Sample records for highly sensitive nested

  1. Development of a highly sensitive one-tube nested real-time PCR for detecting Mycobacterium tuberculosis.

    PubMed

    Choi, Yeonim; Jeon, Bo-Young; Shim, Tae Sun; Jin, Hyunwoo; Cho, Sang-Nae; Lee, Hyeyoung

    2014-12-01

    Rapid, accurate detection of Mycobacterium tuberculosis is crucial in the diagnosis of tuberculosis (TB), but conventional diagnostic methods have limited sensitivity and specificity or are time consuming. A new highly sensitive nucleic acid amplification test, combined nested and real-time polymerase chain reaction (PCR) in a single tube (one-tube nested real-time PCR), was developed for detecting M. tuberculosis, which takes advantage of two PCR techniques, i.e., nested PCR and real-time PCR. One-tube nested real-time PCR was designed to have two sequential reactions with two sets of primers and dual probes for the insertion sequence (IS) 6110 sequence of M. tuberculosis in a single closed tube. The minimum limits of detection of IS6110 real-time PCR and IS6110 one-tube nested real-time PCR were 100 fg/μL and 1 fg/μL of M. tuberculosis DNA, respectively. AdvanSure TB/non-tuberculous mycobacteria (NTM) real-time PCR, IS6110 real-time PCR, and two-tube nested real-time PCR showed 100% sensitivity and 100% specificity for clinical M. tuberculosis isolates and NTM isolates. In comparison, the sensitivities of AdvanSure TB/NTM real-time PCR, single IS6110 real-time PCR, and one-tube nested real-time PCR were 91% (152/167), 94.6% (158/167), and 100% (167/167) for sputum specimens, respectively. In conclusion, IS6110 one-tube nested real-time PCR is useful for detecting M. tuberculosis due to its high sensitivity and simple manipulation. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Novel approach based on one-tube nested PCR and a lateral flow strip for highly sensitive diagnosis of tuberculous meningitis.

    PubMed

    Sun, Yajuan; Chen, Jiajun; Li, Jia; Xu, Yawei; Jin, Hui; Xu, Na; Yin, Rui; Hu, Guohua

    2017-01-01

    Rapid and sensitive detection of Mycobacterium tuberculosis (M. Tb) in cerebrospinal fluid is crucial in the diagnosis of tuberculous meningitis (TBM), but conventional diagnostic technologies have limited sensitivity and specificity or are time-consuming. In this work, a novel, highly sensitive molecular diagnostic method, one-tube nested PCR-lateral flow strip test (OTNPCR-LFST), was developed for detecting M. tuberculosis. This one-tube nested PCR maintains the sensitivity of conventional two-step nested PCR and reduces both the chance of cross-contamination and the time required for analysis. The PCR product was detected by a lateral flow strip assay, which provided a basis for migration of the test to a point-of-care (POC) microfluidic format. The developed assay had an improved sensitivity compared with traditional PCR, and the limit of detection was up to 1 fg DNA isolated from M. tuberculosis. The assay was also specific for M. tuberculosis, and no cross-reactions were found in other non-target bacteria. The application of this technique to clinical samples was successfully evaluated, and OTNPCR-LFST showed 89% overall sensitivity and 100% specificity for TBM patients. This one-tube nested PCR-lateral flow strip assay is useful for detecting M. tuberculosis in TBM due to its rapidity, high sensitivity and simple manipulation.

  3. Novel approach based on one-tube nested PCR and a lateral flow strip for highly sensitive diagnosis of tuberculous meningitis

    PubMed Central

    Sun, Yajuan; Chen, Jiajun; Li, Jia; Xu, Yawei; Jin, Hui; Xu, Na; Yin, Rui

    2017-01-01

    Rapid and sensitive detection of Mycobacterium tuberculosis (M. Tb) in cerebrospinal fluid is crucial in the diagnosis of tuberculous meningitis (TBM), but conventional diagnostic technologies have limited sensitivity and specificity or are time-consuming. In this work, a novel, highly sensitive molecular diagnostic method, one-tube nested PCR-lateral flow strip test (OTNPCR-LFST), was developed for detecting M. tuberculosis. This one-tube nested PCR maintains the sensitivity of conventional two-step nested PCR and reduces both the chance of cross-contamination and the time required for analysis. The PCR product was detected by a lateral flow strip assay, which provided a basis for migration of the test to a point-of-care (POC) microfluidic format. The developed assay had an improved sensitivity compared with traditional PCR, and the limit of detection was up to 1 fg DNA isolated from M. tuberculosis. The assay was also specific for M. tuberculosis, and no cross-reactions were found in other non-target bacteria. The application of this technique to clinical samples was successfully evaluated, and OTNPCR-LFST showed 89% overall sensitivity and 100% specificity for TBM patients. This one-tube nested PCR-lateral flow strip assay is useful for detecting M. tuberculosis in TBM due to its rapidity, high sensitivity and simple manipulation. PMID:29084241

  4. Nested PCR Assay for Eight Pathogens: A Rapid Tool for Diagnosis of Bacterial Meningitis.

    PubMed

    Bhagchandani, Sharda P; Kubade, Sushant; Nikhare, Priyanka P; Manke, Sonali; Chandak, Nitin H; Kabra, Dinesh; Baheti, Neeraj N; Agrawal, Vijay S; Sarda, Pankaj; Mahajan, Parikshit; Ganjre, Ashish; Purohit, Hemant J; Singh, Lokendra; Taori, Girdhar M; Daginawala, Hatim F; Kashyap, Rajpal S

    2016-02-01

    Bacterial meningitis is a dreadful infectious disease with a high mortality and morbidity if remained undiagnosed. Traditional diagnostic methods for bacterial meningitis pose a challenge in accurate identification of pathogen, making prognosis difficult. The present study is therefore aimed to design and evaluate a specific and sensitive nested 16S rDNA genus-based polymerase chain reaction (PCR) assay using clinical cerebrospinal fluid (CSF) for rapid diagnosis of eight pathogens causing the disease. The present work was dedicated to development of an in-house genus specific 16S rDNA nested PCR covering pathogens of eight genera responsible for causing bacterial meningitis using newly designed as well as literature based primers for respective genus. A total 150 suspected meningitis CSF obtained from the patients admitted to Central India Institute of Medical Sciences (CIIMS), India during the period from August 2011 to May 2014, were used to evaluate clinical sensitivity and clinical specificity of optimized PCR assays. The analytical sensitivity and specificity of our newly designed genus-specific 16S rDNA PCR were found to be ≥92%. With such a high sensitivity and specificity, our in-house nested PCR was able to give 100% sensitivity in clinically confirmed positive cases and 100% specificity in clinically confirmed negative cases indicating its applicability in clinical diagnosis. Our in-house nested PCR system therefore can diagnose the accurate pathogen causing bacterial meningitis and therefore be useful in selecting a specific treatment line to minimize morbidity. Results are obtained within 24 h and high sensitivity makes this nested PCR assay a rapid and accurate diagnostic tool compared to traditional culture-based methods.

  5. Sensitivity of nested-PCR for plasmodium detection in pooled whole blood samples and its usefulness to blood donor screening in endemic areas.

    PubMed

    de Freitas, Daniel Roberto Coradi; Gomes, Luciano Teixeira; Fontes, Cor Jesus F; Tauil, Pedro Luiz; Pang, Lorrin W; Duarte, Elisabeth Carmen

    2014-04-01

    Transfusion-transmitted malaria is a severe disease with high fatality rate. Most Brazilian blood banks in the Amazon region perform malaria screening using microscopic examination (thick smears). Since low parasite concentrations are expected in asymptomatic blood donors a high sensitivity test should be used for donor screening. This study determined the sensitivity of a nested-PCR for plasmodium detection in pooled samples. We performed a one-stage criterion validation study with 21 positive samples pooled with samples from ten negative volunteer until three different concentrations were reached (0.33; 0.25; 0.20 parasites/μL - p/μL). Nested PCR was performed as described by Snounou et al. (1993). Sensitivities (and confidence intervals) were determined by stratum of final parasite concentration on the pooled samples. All samples with parasitemia values of 0.33 and 0.25 p/μL had 100% sensitivity (95%CI=86.3-100). One negative result was obtained from a sample with 0.20 p/μL sensitivity=95.2% (95%CI=76.2-99.9). Compared to parasitemia detectable under ideal conditions of thick smear, this nested-PCR in pooled sample was able to detect 40 times more parasites per microliter. Nested-PCR in pooled samples should be considered as a high sensitive alternative to thick smear for donor screening in blood banks at endemic regions. Local authorities need to assess cost:benefit advantages of this method compared to alternatives. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Detection of tumor markers in prostate cancer and comparison of sensitivity between real time and nested PCR.

    PubMed

    Matsuoka, Takayuki; Shigemura, Katsumi; Yamamichi, Fukashi; Fujisawa, Masato; Kawabata, Masato; Shirakawa, Toshiro

    2012-06-27

    The objective of this study is to investigate and compare the sensitivity in conventional PCR, quantitative real time PCR, nested PCR and western blots for detection of prostate cancer tumor markers using prostate cancer (PCa) cells. We performed conventional PCR, quantitative real time PCR, nested PCR, and western blots using 5 kinds of PCa cells. Prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and androgen receptor (AR) were compared for their detection sensitivity by real time PCR and nested PCR. In real time PCR, there was a significant correlation between cell number and the RNA concentration obtained (R(2)=0.9944) for PSA, PSMA, and AR. We found it possible to detect these markers from a single LNCaP cell in both real time and nested PCR. By comparison, nested PCR reached a linear curve in fewer PCR cycles than real time PCR, suggesting that nested PCR may offer PCR results more quickly than real time PCR. In conclusion, nested PCR may offer tumor maker detection in PCa cells more quickly (with fewer PCR cycles) with the same high sensitivity as real time PCR. Further study is necessary to establish and evaluate the best tool for PCa tumor marker detection.

  7. Development of a rapid and sensitive one-step reverse transcription-nested polymerase chain reaction in a single tube using the droplet-polymerase chain reaction machine.

    PubMed

    Yamaguchi, Akemi; Matsuda, Kazuyuki; Sueki, Akane; Taira, Chiaki; Uehara, Masayuki; Saito, Yasunori; Honda, Takayuki

    2015-08-25

    Reverse transcription (RT)-nested polymerase chain reaction (PCR) is a time-consuming procedure because it has several handling steps and is associated with the risk of cross-contamination during each step. Therefore, a rapid and sensitive one-step RT-nested PCR was developed that could be performed in a single tube using a droplet-PCR machine. The K562 BCR-ABL mRNA-positive cell line as well as bone marrow aspirates from 5 patients with chronic myelogenous leukemia (CML) and 5 controls without CML were used. We evaluated one-step RT-nested PCR using the droplet-PCR machine. One-step RT-nested PCR performed in a single tube using the droplet-PCR machine enabled the detection of BCR-ABL mRNA within 40min, which was 10(3)-fold superior to conventional RT nested PCR using three steps in separate tubes. The sensitivity of the one-step RT-nested PCR was 0.001%, with sample reactivity comparable to that of the conventional assay. One-step RT-nested PCR was developed using the droplet-PCR machine, which enabled all reactions to be performed in a single tube accurately and rapidly and with high sensitivity. This one-step RT-nested PCR may be applicable to a wide spectrum of genetic tests in clinical laboratories. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Lizards fail to plastically adjust nesting behavior or thermal tolerance as needed to buffer populations from climate warming.

    PubMed

    Telemeco, Rory S; Fletcher, Brooke; Levy, Ofir; Riley, Angela; Rodriguez-Sanchez, Yesenia; Smith, Colton; Teague, Collin; Waters, Amanda; Angilletta, Michael J; Buckley, Lauren B

    2017-03-01

    Although observations suggest the potential for phenotypic plasticity to allow adaptive responses to climate change, few experiments have assessed that potential. Modeling suggests that Sceloporus tristichus lizards will need increased nest depth, shade cover, or embryonic thermal tolerance to avoid reproductive failure resulting from climate change. To test for such plasticity, we experimentally examined how maternal temperatures affect nesting behavior and embryonic thermal sensitivity. The temperature regime that females experienced while gravid did not affect nesting behavior, but warmer temperatures at the time of nesting reduced nest depth. Additionally, embryos from heat-stressed mothers displayed increased sensitivity to high-temperature exposure. Simulations suggest that critically low temperatures, rather than high temperatures, historically limit development of our study population. Thus, the plasticity needed to buffer this population has not been under selection. Plasticity will likely fail to compensate for ongoing climate change when such change results in novel stressors. © 2016 John Wiley & Sons Ltd.

  9. Development of a Highly Sensitive Nested-PCR Procedure Using a Single Closed Tube for Detection of Erwinia amylovora in Asymptomatic Plant Material

    PubMed Central

    Llop, Pablo; Bonaterra, Anna; Peñalver, Javier; López, María M.

    2000-01-01

    A novel method, which involves a nested PCR in a single closed tube, was developed for the sensitive detection of Erwinia amylovora in plant material. The external and internal primer pairs used had different annealing temperatures and directed the amplification of a specific DNA fragment from plasmid pEA29. The procedure involved two consecutive PCRs, the first of which was performed at a higher annealing temperature that allowed amplification only by the external primer pair. Using pure cultures of E. amylovora, the sensitivity of the nested PCR in one tube was similar to that of a standard nested PCR in two tubes. The specificity and sensitivity were greater than those of standard PCR procedures that used a single primer pair. The presence of inhibitors in plant material, very common in E. amylovora hosts, is overcome with this system in combination with a simple DNA extraction protocol because it eliminates many of the inhibitory compounds. In addition, it needs a very small sample volume (1 μl of DNA extracted). With 83 samples of naturally infected material, this method achieved better results than any other PCR technique: standard PCR detected 55% of positive samples, two-tube nested PCR detected 71% of positive samples, and nested PCR in a single closed tube detected 78% of positive samples. When analyzing asymptomatic plant material, the number of positive samples detected by the developed nested PCR was also the highest, compared with the PCR protocols indicated previously (17, 20, and 25% of 251 samples analyzed, respectively). This method is proposed for the detection of endophytic and epiphytic populations of E. amylovora in epidemiological studies and for routine use in quarantine surveys, due to its high sensitivity, specificity, speed, and simplicity. PMID:10788384

  10. Evaluation of highly conserved hsp65-specific nested PCR primers for diagnosing Mycobacterium tuberculosis.

    PubMed

    Priyadarshini, P; Tiwari, K; Das, A; Kumar, D; Mishra, M N; Desikan, P; Nath, G

    2017-02-01

    To evaluate the sensitivity and specificity of a new nested set of primers designed for the detection of Mycobacterium tuberculosis complex targeting a highly conserved heat shock protein gene (hsp65). The nested primers were designed using multiple sequence alignment assuming the nucleotide sequence of the M. tuberculosis H37Rv hsp65 genome as base. Multidrug-resistant Mycobacterium species along with other non-mycobacterial and fungal species were included to evaluate the specificity of M. tuberculosis hsp65 gene-specific primers. The sensitivity of the primers was determined using serial 10-fold dilutions, and was 100% as shown by the bands in the case of M. tuberculosis complex. None of the other non M. tuberculosis complex bacterial and fungal species yielded any band on nested polymerase chain reaction (PCR). The first round of amplification could amplify 0.3 ng of the template DNA, while nested PCR could detect 0.3 pg. The present hsp65-specific primers have been observed to be sensitive, specific and cost-effective, without requiring interpretation of biochemical tests, real-time PCR, sequencing or high-performance liquid chromatography. These primer sets do not have the drawbacks associated with those protocols that target insertion sequence 6110, 16S rDNA, rpoB, recA and MPT 64.

  11. Diagnostic value of nested-PCR for identification of Malassezia species in dandruff

    NASA Astrophysics Data System (ADS)

    Jusuf, N. K.; Nasution, T. A.; Ullyana, S.

    2018-03-01

    Dandruff or pityriasis simplex is a condition of abnormal occurrence of formation of yellowish white scales from the scalp. Many factors play a role in the pathogenesis of dandruff, i.e.colonization of Malassezia species. Examination of Malassezia species previously done by culture as the gold standard. However, there are various difficulties in doing the culture. Identification method with anested-polymerase chain reaction (nested-PCR) is expected to provide quickly and easily detected. This study aimedto determine the diagnostic value of nested-PCR in the identification of Malassezia species in dandruff. From 21 subjects, scales from the scalp were taken and sent to the laboratory for nested-PCR identification. Statistical analysis of diagnostic test carried out to determine sensitivity, specificity, positive predictive value, and negative predictive value. The results showed nested-PCR detected 10 sample (47.6%) positive for Malassezia species consist of M. sympodialis (23.8%); M. slooffiae (9.5%); M. furfur (4.8%); M. globosa and M. furfur (4.8%); and M. restricta and M. sympodialis (4.8%). Detection of Malassezia species by nested-PCR has 100% in sensitivity whereas the specificity was 55%. Nested-PCR test has high sensitivity. Therefore nested-PCR may be considered for a faster and simpler alternative examination in identification for Malassezia species in dandruff.

  12. A highly sensitive single-tube nested PCR assay for the detection of Pineapple mealybug wilt associated virus-2 (PMWaV-2)

    USDA-ARS?s Scientific Manuscript database

    An assay was developed for the detection of Pineapple mealybug wilt associated virus-2 (PMWaV-2), an important factor in the etiology of mealybug wilt of pineapple. The assay combines reverse transcription of RNA isolated from pineapple with a specific and very sensitive, single, closed-tube nested ...

  13. Establishment of a nested-ASP-PCR method to determine the clarithromycin resistance of Helicobacter pylori.

    PubMed

    Luo, Xiao-Feng; Jiao, Jian-Hua; Zhang, Wen-Yue; Pu, Han-Ming; Qu, Bao-Jin; Yang, Bing-Ya; Hou, Min; Ji, Min-Jun

    2016-07-07

    To investigate clarithromycin resistance positions 2142, 2143 and 2144 of the 23SrRNA gene in Helicobacter pylori (H. pylori) by nested-allele specific primer-polymerase chain reaction (nested-ASP-PCR). The gastric tissue and saliva samples from 99 patients with positive results of the rapid urease test (RUT) were collected. The nested-ASP-PCR method was carried out with the external primers and inner allele-specific primers corresponding to the reference strain and clinical strains. Thirty gastric tissue and saliva samples were tested to determine the sensitivity of nested-ASP-PCR and ASP-PCR methods. Then, clarithromycin resistance was detected for 99 clinical samples by using different methods, including nested-ASP-PCR, bacterial culture and disk diffusion. The nested-ASP-PCR method was successfully established to test the resistance mutation points 2142, 2143 and 2144 of the 23SrRNA gene of H. pylori. Among 30 samples of gastric tissue and saliva, the H. pylori detection rate of nested-ASP-PCR was 90% and 83.33%, while the detection rate of ASP-PCR was just 63% and 56.67%. Especially in the saliva samples, nested-ASP-PCR showed much higher sensitivity in H. pylori detection and resistance mutation rates than ASP-PCR. In the 99 RUT-positive gastric tissue and saliva samples, the H. pylori-positive detection rate by nested-ASP-PCR was 87 (87.88%) and 67 (67.68%), in which there were 30 wild-type and 57 mutated strains in gastric tissue and 22 wild-type and 45 mutated strains in saliva. Genotype analysis showed that three-points mixed mutations were quite common, but different resistant strains were present in gastric mucosa and saliva. Compared to the high sensitivity shown by nested-ASP-PCR, the positive detection of bacterial culture with gastric tissue samples was 50 cases, in which only 26 drug-resistant strains were found through analyzing minimum inhibitory zone of clarithromycin. The nested-ASP-PCR assay showed higher detection sensitivity than ASP-PCR and drug sensitivity testing, which could be performed to evaluate clarithromycin resistance of H. pylori.

  14. Establishment of a nested-ASP-PCR method to determine the clarithromycin resistance of Helicobacter pylori

    PubMed Central

    Luo, Xiao-Feng; Jiao, Jian-Hua; Zhang, Wen-Yue; Pu, Han-Ming; Qu, Bao-Jin; Yang, Bing-Ya; Hou, Min; Ji, Min-Jun

    2016-01-01

    AIM: To investigate clarithromycin resistance positions 2142, 2143 and 2144 of the 23SrRNA gene in Helicobacter pylori (H. pylori) by nested-allele specific primer-polymerase chain reaction (nested-ASP-PCR). METHODS: The gastric tissue and saliva samples from 99 patients with positive results of the rapid urease test (RUT) were collected. The nested-ASP-PCR method was carried out with the external primers and inner allele-specific primers corresponding to the reference strain and clinical strains. Thirty gastric tissue and saliva samples were tested to determine the sensitivity of nested-ASP-PCR and ASP-PCR methods. Then, clarithromycin resistance was detected for 99 clinical samples by using different methods, including nested-ASP-PCR, bacterial culture and disk diffusion. RESULTS: The nested-ASP-PCR method was successfully established to test the resistance mutation points 2142, 2143 and 2144 of the 23SrRNA gene of H. pylori. Among 30 samples of gastric tissue and saliva, the H. pylori detection rate of nested-ASP-PCR was 90% and 83.33%, while the detection rate of ASP-PCR was just 63% and 56.67%. Especially in the saliva samples, nested-ASP-PCR showed much higher sensitivity in H. pylori detection and resistance mutation rates than ASP-PCR. In the 99 RUT-positive gastric tissue and saliva samples, the H. pylori-positive detection rate by nested-ASP-PCR was 87 (87.88%) and 67 (67.68%), in which there were 30 wild-type and 57 mutated strains in gastric tissue and 22 wild-type and 45 mutated strains in saliva. Genotype analysis showed that three-points mixed mutations were quite common, but different resistant strains were present in gastric mucosa and saliva. Compared to the high sensitivity shown by nested-ASP-PCR, the positive detection of bacterial culture with gastric tissue samples was 50 cases, in which only 26 drug-resistant strains were found through analyzing minimum inhibitory zone of clarithromycin. CONCLUSION: The nested-ASP-PCR assay showed higher detection sensitivity than ASP-PCR and drug sensitivity testing, which could be performed to evaluate clarithromycin resistance of H. pylori. PMID:27433095

  15. Use of a nested PCR-enzyme immunoassay with an internal control to detect Chlamydophila psittaci in turkeys

    PubMed Central

    Van Loock, Marnix; Verminnen, Kristel; Messmer, Trudy O; Volckaert, Guido; Goddeeris, Bruno M; Vanrompay, Daisy

    2005-01-01

    Background Laboratory diagnosis of Chlamydophila psittaci, an important turkey respiratory pathogen, is difficult. To facilitate the diagnosis, a nested PCR-enzyme immunoassay (PCR-EIA) was developed to detect the Cp. psittaci outer membrane protein A (ompA) gene in pharyngeal swabs. Methods The fluorescein-biotin labelled PCR products were immobilized on streptavidin-coated microtiter plates and detected with anti-fluorescein peroxidase conjugate and a colorimetric substrate. An internal inhibition control was included to rule out the presence of inhibitors of DNA amplification. The diagnostic value of the ompA nested PCR-EIA in comparison to cell culture and a 16S-rRNA based nested PCR was assessed in pharyngeal turkey swabs from 10 different farms experiencing respiratory disease. Results The sensitivity of the nested PCR-EIA was established at 0.1 infection forming units (IFU). Specificity was 100%. The ompA nested PCR-EIA was more sensitive than the 16S-rRNA based nested PCR and isolation, revealing 105 out of 200 (52.5%) positives against 13 and 74 for the latter two tests, respectively. Twenty-nine (23.8%) out of 122 ompA PCR-EIA negatives showed the presence of inhibitors of DNA amplification, although 27 of them became positive after diluting (1/10) the specimens in PCR buffer or after phenol-chloroform extraction and subsequent ethanol precipitation. Conclusion The present study stresses the need for an internal control to confirm PCR true-negatives and demonstrates the high prevalence of chlamydiosis in Belgian turkeys and its potential zoonotic risk. The ompA nested PCR-EIA described here is a rapid, highly sensitive and specific diagnostic assay and will help to facilitate the diagnosis of Cp. psittaci infections in both poultry and man. PMID:16185353

  16. Detection of low plasma estradiol concentrations in nesting green turtles (Chelonia mydas) by HPLC/Ms-Ms.

    PubMed

    Mahmoud, I Y; Alkindi, A Y; Khan, T; Al-Bahry, S N

    2011-03-01

    In previous studies on nesting green turtles under natural conditions from different geographical regions, 17-β-estradiol (E(2) ) was either undetectable or detected at very low levels. RIA and other related techniques were not sensitive enough to measure low E(2) values in the green turtles. In this study, a sensitive method was used in detecting low hormone concentrations: high performance liquid chromatography with tandem quadruple mass spectrometry (HPLC-MS/MS). Using this technique, estradiol for the first time was detected in nesting green turtles during the peak season (June-October) at Ras Al-Hadd Reserve, Oman. The E(2) values recorded from this study were the highest ever recorded from nesting green turtles in any geographical region, but the levels did not vary significantly throughout different phases of nesting. The presence of E(2) during nesting presumably plays a role in the physiology and behavior of this species. Ras Al-Hadd hosts one of the largest nesting populations of green turtles in the world, and an understanding of their nesting patterns may be of value in conservation and management programs for this endangered species. Copyright © 2011 Wiley-Liss, Inc., A Wiley Company.

  17. Rapid and sensitive detection of mink circovirus by recombinase polymerase amplification.

    PubMed

    Ge, Junwei; Shi, Yunjia; Cui, Xingyang; Gu, Shanshan; Zhao, Lili; Chen, Hongyan

    2018-06-01

    To date, the pathogenic role of mink circovirus (MiCV) remains unclear, and its prevalence and economic importance are unknown. Therefore, a rapid and sensitive molecular diagnosis is necessary for disease management and epidemiological surveillance. However, only PCR methods can identify MiCV infection at present. In this study, we developed a nested PCR and established a novel recombinase polymerase amplification (RPA) assay for MiCV detection. Sensitivity analysis showed that the detection limit of nested PCR and RPA assay was 10 1 copies/reaction, and these methods were more sensitive than conventional PCR, which has a detection limit of 10 5 copies/reaction. The RPA assay had no cross-reactivity with other related viral pathogens, and amplification was completed in less than 20 min with a simple device. Further assessment of clinical samples showed that the two assays were accurate in identifying positive and negative conventional PCR samples. The detection rate of MiCV by the RPA assay in clinical samples was 38.09%, which was 97% consistent with that by the nested PCR. The developed nested PCR is a highly sensitive tool for practical use, and the RPA assay is a simple, sensitive, and potential alternative method for rapid and accurate MiCV diagnosis. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Molecular diagnosis of lyssaviruses and sequence comparison of Australian bat lyssavirus samples.

    PubMed

    Foord, A J; Heine, H G; Pritchard, L I; Lunt, R A; Newberry, K M; Rootes, C L; Boyle, D B

    2006-07-01

    To evaluate and implement molecular diagnostic tests for the detection of lyssaviruses in Australia. A published hemi-nested reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of all lyssavirus genotypes was modified to a fully nested RT-PCR format and compared with the original assay. TaqMan assays for the detection of Australian bat lyssavirus (ABLV) were compared with both the nested and hemi-nested RT-PCR assays. The sequences of RT-PCR products were determined to assess sequence variations of the target region (nucleocapsid gene) in samples of ABLV originating from different regions. The nested RT-PCR assay was highly analytically specific, and at least as analytically sensitive as the hemi-nested assay. The TaqMan assays were highly analytically specific and more analytically sensitive than either RT-PCR assay, with a detection level of approximately 10 genome equivalents per microl. Sequence of the first 544 nucleotides of the nucleocapsid protein coding sequence was obtained from all samples of ABLV received at Australian Animal Health Laboratory during the study period. The nested RT-PCR provided a means for molecular diagnosis of all tested genotypes of lyssavirus including classical rabies virus and Australian bat lyssavirus. The published TaqMan assay proved to be superior to the RT-PCR assays for the detection of ABLV in terms of analytical sensitivity. The TaqMan assay would also be faster and cross contamination is less likely. Nucleotide sequence analyses of samples of ABLV from a wide geographical range in Australia demonstrated the conserved nature of this region of the genome and therefore the suitability of this region for molecular diagnosis.

  19. Rapid direct identification of Cryptococcus neoformans from pigeon droppings by nested PCR using CNLAC1 gene.

    PubMed

    Chae, H S; Park, G N; Kim, S H; Jo, H J; Kim, J T; Jeoung, H Y; An, D J; Kim, N H; Shin, B W; Kang, Y I; Chang, K S

    2012-08-01

    Isolation and identification of Cryptococcus neoformans and pathogenic yeast-like fungi from pigeon droppings has been taken for a long time and requires various nutrients for its growth. In this study, we attempted to establish a rapid direct identification method of Cr. neoformans from pigeon dropping samples by nested-PCR using internal transcribed spacer (ITS) CAP64 and CNLAC1 genes, polysaccharide capsule gene and laccase-associated gene to produce melanin pigment, respectively, which are common genes of yeasts. The ITS and CAP64 genes were amplified in all pathogenic yeasts, but CNLAC1 was amplified only in Cr. neoformans. The ITS gene was useful for yeast genotyping depending on nucleotide sequence. Homology of CAP64 genes among the yeasts were very high. The specificity of PCR using CNLAC1 was demonstrated in Cr. neoformans environmental strains but not in other yeast-like fungi. The CNLAC1 gene was detected in 5 serotypes of Cr. neoformans. The nested-PCR amplified up to 10(-11) μg of the genomic DNA and showed high sensitivity. All pigeon droppings among 31 Cr. neoformans-positive samples were positive and all pigeon droppings among 348 Cr. neoformans-negative samples were negative by the direct nested-PCR. In addition, after primary enrichment of pigeon droppings in Sabouraud dextrose broth, all Cr. neoformans-negative samples were negative by the nested-PCR, which showed high specificity. The nested-PCR showed high sensitivity without culture of pigeon droppings. Nested-PCR using CNLAC1 provides a rapid and reliable molecular diagnostic method to overcome weak points such as long culture time of many conventional methods.

  20. Evaluation of hsp65 Nested PCR-Restriction Analysis (PRA) for Diagnosing Tuberculosis in a High Burden Country

    PubMed Central

    Macente, Sara; Fujimura Leite, Clarice Queico; Santos, Adolfo Carlos Barreto; Siqueira, Vera Lúcia Dias; Machado, Luzia Neri Cosmo; Marcondes, Nadir Rodrigues; Hirata, Mario Hiroyuki; Hirata, Rosário Dominguez Crespo

    2013-01-01

    Current study evaluated the hsp65 Nested PCR Restriction Fragment Length Polymorphism Analysis (hsp65 Nested PCR-PRA) to detect and identify Mycobacterium tuberculosis complex directly in clinical samples for a rapid and specific diagnosis of tuberculosis (TB). hsp65 Nested PCR-PRA was applied directly to 218 clinical samples obtained from 127 patients suspected of TB or another mycobacterial infection from July 2009 to July 2010. The hsp65 Nested PCR-PRA showed 100% sensitivity and 95.0 and 93.1% specificity in comparison with culture and microscopy (acid fast bacillus smear), respectively. hsp65 Nested PCR-PRA was shown to be a fast and reliable assay for diagnosing TB, which may contribute towards a fast diagnosis that could help the selection of appropriate chemotherapeutic and early epidemiological management of the cases which are of paramount importance in a high TB burden country. PMID:24260739

  1. Development of an In-House Multiplex Nested RT-PCR Method for Detecting Acute HIV-1 Infection in High Risk Populations.

    PubMed

    Liu, Zhiying; Li, Wei; Xu, Meng; Sheng, Bo; Yang, Zixuan; Jiao, Yanmei; Zhang, Tong; Mou, Danlei; Chen, Dexi; Wu, Hao

    2015-01-01

    The detection of acute HIV infection (AHI) among high risk populations can help reduce secondary transmission of HIV. The nucleic acid testing (NAT) can shorten the test window period by up to 7-12 days. In this study, we describe an in-house NAT based on the multiplex nested RT-PCR method to detect the HIV RNA. We also evaluated it in a high risk cohort in Beijing. Four primer pairs were designed and evaluated for the detection of different HIV-1 subtypes in group M. Multiplex RT-PCR and nested PCR were performed. The sensitivity, specialty, primers compatibility among HIV subtypes were evaluated simultaneously. In an MSM cohort in Beijing during a 3-year period, a total of 11,808 blood samples that were negative by ELISA or indeterminate by Western blot were analyzed by this multiplex nested RT-PCR with pooling strategy. The multiplex nested RT-PCR was successfully applied for the detection of at least six HIV-1 subtypes. The sensitivity was 40 copies/ml and the specificity was 100%. A total of 29 people were tested HIV-1 positive with acute infection in a MSM cohort of Beijing during a 3 years period. This multiplex nested RT-PCR provides a useful tool for the rapid detection of acute HIV-1 infection. When used in combination with the 3(rd) generation ELISA, it can improve the detection rate of HIV infection, especially in the source limited regions.

  2. Towards high-throughput molecular detection of Plasmodium: new approaches and molecular markers

    PubMed Central

    Steenkeste, Nicolas; Incardona, Sandra; Chy, Sophy; Duval, Linda; Ekala, Marie-Thérèse; Lim, Pharath; Hewitt, Sean; Sochantha, Tho; Socheat, Doung; Rogier, Christophe; Mercereau-Puijalon, Odile; Fandeur, Thierry; Ariey, Frédéric

    2009-01-01

    Background Several strategies are currently deployed in many countries in the tropics to strengthen malaria control toward malaria elimination. To measure the impact of any intervention, there is a need to detect malaria properly. Mostly, decisions still rely on microscopy diagnosis. But sensitive diagnosis tools enabling to deal with a large number of samples are needed. The molecular detection approach offers a much higher sensitivity, and the flexibility to be automated and upgraded. Methods Two new molecular methods were developed: dot18S, a Plasmodium-specific nested PCR based on the 18S rRNA gene followed by dot-blot detection of species by using species-specific probes and CYTB, a Plasmodium-specific nested PCR based on cytochrome b gene followed by species detection using SNP analysis. The results were compared to those obtained with microscopic examination and the "standard" 18S rRNA gene based nested PCR using species specific primers. 337 samples were diagnosed. Results Compared to the microscopy the three molecular methods were more sensitive, greatly increasing the estimated prevalence of Plasmodium infection, including P. malariae and P. ovale. A high rate of mixed infections was uncovered with about one third of the villagers infected with more than one malaria parasite species. Dot18S and CYTB sensitivity outranged the "standard" nested PCR method, CYTB being the most sensitive. As a consequence, compared to the "standard" nested PCR method for the detection of Plasmodium spp., the sensitivity of dot18S and CYTB was respectively 95.3% and 97.3%. Consistent detection of Plasmodium spp. by the three molecular methods was obtained for 83% of tested isolates. Contradictory results were mostly related to detection of Plasmodium malariae and Plasmodium ovale in mixed infections, due to an "all-or-none" detection effect at low-level parasitaemia. Conclusion A large reservoir of asymptomatic infections was uncovered using the molecular methods. Dot18S and CYTB, the new methods reported herein are highly sensitive, allow parasite DNA extraction as well as genus- and species-specific diagnosis of several hundreds of samples, and are amenable to high-throughput scaling up for larger sample sizes. Such methods provide novel information on malaria prevalence and epidemiology and are suited for active malaria detection. The usefulness of such sensitive malaria diagnosis tools, especially in low endemic areas where eradication plans are now on-going, is discussed in this paper. PMID:19402894

  3. Nested-PCR and a new ELISA-based NovaLisa test kit for malaria diagnosis in an endemic area of Thailand.

    PubMed

    Thongdee, Pimwan; Chaijaroenkul, Wanna; Kuesap, Jiraporn; Na-Bangchang, Kesara

    2014-08-01

    Microscopy is considered as the gold standard for malaria diagnosis although its wide application is limited by the requirement of highly experienced microscopists. PCR and serological tests provide efficient diagnostic performance and have been applied for malaria diagnosis and research. The aim of this study was to investigate the diagnostic performance of nested PCR and a recently developed an ELISA-based new rapid diagnosis test (RDT), NovaLisa test kit, for diagnosis of malaria infection, using microscopic method as the gold standard. The performance of nested-PCR as a malaria diagnostic tool is excellent with respect to its high accuracy, sensitivity, specificity, and ability to discriminate Plasmodium species. The sensitivity and specificity of nested-PCR compared with the microscopic method for detection of Plasmodium falciparum, Plasmodium vivax, and P. falciparum/P. vivax mixed infection were 71.4 vs 100%, 100 vs 98.7%, and 100 vs 95.0%, respectively. The sensitivity and specificity of the ELISA-based NovaLisa test kit compared with the microscopic method for detection of Plasmodium genus were 89.0 vs 91.6%, respectively. NovaLisa test kit provided comparable diagnostic performance. Its relatively low cost, simplicity, and rapidity enables large scale field application.

  4. [Sensitivity and specificity of nested PCR pyrosequencing in hepatitis B virus drug resistance gene testing].

    PubMed

    Sun, Shumei; Zhou, Hao; Zhou, Bin; Hu, Ziyou; Hou, Jinlin; Sun, Jian

    2012-05-01

    To evaluate the sensitivity and specificity of nested PCR combined with pyrosequencing in the detection of HBV drug-resistance gene. RtM204I (ATT) mutant and rtM204 (ATG) nonmutant plasmids mixed at different ratios were detected for mutations using nested-PCR combined with pyrosequencing, and the results were compared with those by conventional PCR pyrosequencing to analyze the linearity and consistency of the two methods. Clinical specimens with different viral loads were examined for drug-resistant mutations using nested PCR pyrosequencing and nested PCR combined with dideoxy sequencing (Sanger) for comparison of the detection sensitivity and specificity. The fitting curves demonstrated good linearity of both conventional PCR pyrosequencing and nested PCR pyrosequencing (R(2)>0.99, P<0.05). Nested PCR showed a better consistency with the predicted value than conventional PCR, and was superior to conventional PCR for detection of samples containing 90% mutant plasmid. In the detection of clinical specimens, Sanger sequencing had a significantly lower sensitivity than nested PCR pyrosequencing (92% vs 100%, P<0.01). The detection sensitivity of Sanger sequencing varied with the viral loads, especially in samples with low viral copies (HBV DNA ≤3log10 copies/ml), where the sensitivity was 78%, significantly lower than that of pyrosequencing (100%, P<0.01). Neither of the two methods yielded positive results for the negative control samples, suggesting their good specificity. Compared with nested PCR and Sanger sequencing method, nested PCR pyrosequencing has a higher sensitivity especially in clinical specimens with low viral copies, which can be important for early detection of HBV mutant strains and hence more effective clinical management.

  5. Monitoring and improving the sensitivity of dengue nested RT-PCR used in longitudinal surveillance in Thailand.

    PubMed

    Klungthong, Chonticha; Manasatienkij, Wudtichai; Phonpakobsin, Thipwipha; Chinnawirotpisan, Piyawan; Rodpradit, Prinyada; Hussem, Kittinun; Thaisomboonsuk, Butsaya; Ong-ajchaowlerd, Prapapun; Nisalak, Ananda; Kalayanarooj, Siripen; Buddhari, Darunee; Gibbons, Robert V; Jarman, Richard G; Yoon, In-Kyu; Fernandez, Stefan

    2015-02-01

    AFRIMS longitudinal dengue surveillance in Thailand depends on the nested RT-PCR and the dengue IgM/IgG ELISA. To examine and improve the sensitivity of the nested RT-PCR using a panel of archived samples collected during dengue surveillance. A retrospective analysis of 16,454 dengue IgM/IgG ELISA positive cases collected between 2000 and 2013 was done to investigate the sensitivity of the nested RT-PCR. From these cases, 318 acute serum specimens or extracted RNA, previously found to be negative by the nested RT-PCR, were tested using TaqMan real-time RT-PCR (TaqMan rRT-PCR). To improve the sensitivity of nested RT-PCR, we designed a new primer based on nucleotide sequences from contemporary strains found to be positive by the TaqMan rRT-PCR. Sensitivity of the new nested PCR was calculated using a panel of 87 samples collected during 2011-2013. The percentage of dengue IgM/IgG ELISA positive cases that were negative by the nested RT-PCR varied from 17% to 42% for all serotypes depending on the year. Using TaqMan rRT-PCR, dengue RNA was detected in 194 (61%) of the 318 acute sera or extracted RNA previously found to be negative by the nested RT-PCR. The newly designed DENV-1 specific primer increased the sensitivity of DENV-1 detection by the nested RT-PCR from 48% to 88%, and of all 4 serotypes from 73% to 87%. These findings demonstrate the impact of genetic diversity and signal erosion on the sensitivity of PCR-based methods. Published by Elsevier B.V.

  6. Development of a highly sensitive real-time nested RT-PCR assay in a single closed tube for detection of enterovirus 71 in hand, foot, and mouth disease.

    PubMed

    Niu, Peihua; Qi, Shunxiang; Yu, Benzhang; Zhang, Chen; Wang, Ji; Li, Qi; Ma, Xuejun

    2016-11-01

    Enterovirus 71 (EV71) is one of the major causative agents of outbreaks of hand, foot, and mouth disease (HFMD). A commercial TaqMan probe-based real-time PCR assay has been widely used for the differential detection of EV71 despite its relatively high cost and failure to detect samples with a low viral load (Ct value > 35). In this study, a highly sensitive real-time nested RT-PCR (RTN RT-PCR) assay in a single closed tube for detection of EV71 in HFMD was developed. The sensitivity and specificity of this assay were evaluated using a reference EV71 stock and a panel of controls consisting of coxsackievirus A16 (CVA16) and common respiratory viruses, respectively. The clinical performance of this assay was evaluated and compared with those of a commercial TaqMan probe-based real-time PCR (qRT-PCR) assay and a traditional two-step nested RT-PCR assay. The limit of detection for the RTN RT-PCR assay was 0.01 TCID50/ml, with a Ct value of 38.3, which was the same as that of the traditional two-step nested RT-PCR assay and approximately tenfold lower than that of the qRT-PCR assay. When testing the reference strain EV71, this assay showed favorable detection reproducibility and no obvious cross-reactivity. The testing results of 100 clinical throat swabs from HFMD-suspected patients revealed that 41 samples were positive for EV71 by both RTN RT-PCR and traditional two-step nested RT-PCR assays, whereas only 29 were EV71 positive by qRT-PCR assay.

  7. Estimating raptor nesting success: old and new approaches

    USGS Publications Warehouse

    Brown, Jessi L.; Steenhof, Karen; Kochert, Michael N.; Bond, Laura

    2013-01-01

    Studies of nesting success can be valuable in assessing the status of raptor populations, but differing monitoring protocols can present unique challenges when comparing populations of different species across time or geographic areas. We used large datasets from long-term studies of 3 raptor species to compare estimates of apparent nest success (ANS, the ratio of successful to total number of nesting attempts), Mayfield nesting success, and the logistic-exposure model of nest survival. Golden eagles (Aquila chrysaetos), prairie falcons (Falco mexicanus), and American kestrels (F. sparverius) differ in their breeding biology and the methods often used to monitor their reproduction. Mayfield and logistic-exposure models generated similar estimates of nesting success with similar levels of precision. Apparent nest success overestimated nesting success and was particularly sensitive to inclusion of nesting attempts discovered late in the nesting season. Thus, the ANS estimator is inappropriate when exact point estimates are required, especially when most raptor pairs cannot be located before or soon after laying eggs. However, ANS may be sufficient to assess long-term trends of species in which nesting attempts are highly detectable.

  8. Molecular diagnosis of strongyloidiasis in a population of an endemic area through nested-PCR.

    PubMed

    Sharifdini, Meysam; Keyhani, Amir; Eshraghian, Mohammad Reza; Beigom Kia, Eshrat

    2018-01-01

    This study is aimed to diagnose and analyze strongyloidiasis in a population of an endemic area of Iran using nested-PCR, coupled with parasitological methods. Screening of strongyloidiasis infected people using reliable diagnostic techniques are essential to decrease the mortality and morbidity associated with this infection. Molecular methods have been proved to be highly sensitive and specific for detection of Strongyloides stercoralis in stool samples. A total of 155 fresh single stool samples were randomly collected from residents of north and northwest of Khouzestan Province, Iran. All samples were examined by parasitological methods including formalin-ether concentration and nutrient agar plate culture, and molecular method of nested-PCR. Infections with S. stercoralis were analyzed according to demographic criteria. Based on the results of nested-PCR method 15 cases (9.7%) were strongyloidiasis positive. Nested-PCR was more sensitive than parasitological techniques on single stool sampling. Elderly was the most important population index for higher infectivity with S. stercoralis . In endemic areas of S. stercoralis , old age should be considered as one of the most important risk factors of infection, especially among the immunosuppressed individuals.

  9. Immunocapture reverse transcription-polymerase chain reaction combined with nested PCR greatly increases the detection of Prunus necrotic ring spot virus in the peach.

    PubMed

    Helguera, P R; Taborda, R; Docampo, D M; Ducasse, D A

    2001-06-01

    A detection system based on nested PCR after IC-RT-PCR (IC-RT-PCR-Nested PCR) was developed to improve indexing of Prunus necrotic ringspot virus in peach trees. Inhibitory effects and inconsistencies of the standard IC-RT-PCR were overcome by this approach. IC-RT-PCR-Nested PCR improved detection by three orders of magnitude compared with DAS-ELISA for the detection of PNRSV in leaves. Several different tissues were evaluated and equally consistent results were observed. The main advantages of the method are its consistency, high sensitivity and easy application in quarantine programs.

  10. A novel and highly sensitive real-time nested RT-PCR assay in a single closed tube for detection of enterovirus.

    PubMed

    Shen, Xin-Xin; Qiu, Fang-Zhou; Zhao, Huai-Long; Yang, Meng-Jie; Hong, Liu; Xu, Song-Tao; Zhou, Shuai-Feng; Li, Gui-Xia; Feng, Zhi-Shan; Ma, Xue-Jun

    2018-03-01

    The sensitivity of qRT-PCR assay is not adequate for the detection of the samples with lower viral load, particularly in the cerebrospinal fluid (CSF) of patients. Here, we present the development of a highly sensitive real-time nested RT-PCR (RTN RT-PCR) assay in a single closed tube for detection of human enterovirus (HEV). The clinical performance of both RTN RT-PCR and qRT-PCR was also tested and compared using 140 CSF and fecal specimens. The sensitivities of RTN RT-PCR assay for EV71, Coxsackievirus A (CVA)16, CVA6 and CVA10 achieved 10 -8 dilution with a corresponding Ct value of 38.20, 36.45, 36.75, and 36.45, respectively, which is equal to traditional two-step nested RT-PCR assay and approximately 2-10-fold lower than that of qRT-PCR assay. The specificity of RTN RT-PCR assay was extensively analyzed insilico and subsequently verified using the reference isolates and clinical samples. Sixteen qRT-PCR-negative samples were detected by RTN RT-PCR and a variety of enterovirus serotypes was identified by sequencing of inner PCR products. We conclude RTN RT-PCR is more sensitive than qRT-PCR for the detection of HEV in clinical samples. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. The Diagnostic Utility of Bact/ALERT and Nested PCR in the Diagnosis of Tuberculous Meningitis.

    PubMed

    Sastry, Apurba Sankar; Bhat K, Sandhya; Kumudavathi

    2013-01-01

    The early laboratory diagnosis of Tuberculous Meningitis (TBM) is crucial, to start the antitubercular chemotherapy and to prevent its complications. However, the conventional methods are either less sensitive or time consuming. Hence, the diagnostic potentials of BacT/ALERT and Polymerase Chain Reaction (PCR) was evaluated in this study. The study group comprised of 62 cases and 33 controls. The cases were divided according to Ahuja's criteria into the confirmed (two cases), highly probable (19 cases), probable (26 cases) and the possible (15 cases) subgroups. Ziehl Neelsen's (ZN) and Auramine Phenol (AP) staining, Lowenstein Jensen (LJ) medium culture, BacT/ALERT and nested Polymerase Chain Reaction (PCR) which targeted IS6110 were carried out on all the patients. The sensitivity of the LJ culture was 3.22%. BacT/ALERT showed a sensitivity and a specificity of 25.80% and 100% and those of nested PCR were found to be 40.32% and 96.97% respectively. The mean detection time of growth of the LJ culture was 31.28 days, whereas that of BacT/ALERT was 20.68 days. The contamination rate in the LJ culture and BacT/ALERT were 7.2% and 5.8% respectively. Nested PCR was found to be more sensitive, followed by BacT/ALERT as compared to the LJ culture and smear microscopy. As both false negative and false positive results have been reported for nested PCR, so it should not be used alone as a criterion for initiating or terminating the therapy, but it should be supported by clinical, radiological, cytological and other microbiological findings.

  12. Comparison of Four Human Papillomavirus Genotyping Methods: Next-generation Sequencing, INNO-LiPA, Electrochemical DNA Chip, and Nested-PCR.

    PubMed

    Nilyanimit, Pornjarim; Chansaenroj, Jira; Poomipak, Witthaya; Praianantathavorn, Kesmanee; Payungporn, Sunchai; Poovorawan, Yong

    2018-03-01

    Human papillomavirus (HPV) infection causes cervical cancer, thus necessitating early detection by screening. Rapid and accurate HPV genotyping is crucial both for the assessment of patients with HPV infection and for surveillance studies. Fifty-eight cervicovaginal samples were tested for HPV genotypes using four methods in parallel: nested-PCR followed by conventional sequencing, INNO-LiPA, electrochemical DNA chip, and next-generation sequencing (NGS). Seven HPV genotypes (16, 18, 31, 33, 45, 56, and 58) were identified by all four methods. Nineteen HPV genotypes were detected by NGS, but not by nested-PCR, INNO-LiPA, or electrochemical DNA chip. Although NGS is relatively expensive and complex, it may serve as a sensitive HPV genotyping method. Because of its highly sensitive detection of multiple HPV genotypes, NGS may serve as an alternative for diagnostic HPV genotyping in certain situations. © The Korean Society for Laboratory Medicine

  13. Determination of an effective sampling regime to detect salmonella enteritidis in the environment of poultry units.

    PubMed

    Davies, R H; Wray, C

    1996-05-01

    A study of the dissemination of Salmonella enteritidis in the poultry breeder industry in the UK showed that the choice of sites for sampling the environment of occupied houses and empty houses which had been disinfected after depopulation had a significant influence on the outcome. Increased isolation rates could be achieved by sampling nest box floors and dust in open slave feed hoppers in occupied poultry houses. Nest box floors were the most sensitive sites for detection of residual environmental contamination in poultry houses where enrofloxacin treatment had been used. Floor sweepings, nest box floors, slave feed hoppers, hydrated wall fabric junctions and high beams and pipes were the most sensitive sample sites in cleansed and disinfected poultry houses. The use of universal disinfectant neutralisers gave good results in laboratory trials but appeared to reduce the isolation rate from field samples.

  14. Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé.

    PubMed

    Lee, Pei-Wen; Ji, Dar-Der; Liu, Chia-Tai; Rampao, Herodes S; do Rosario, Virgilio E; Lin, I-Feng; Shaio, Men-Fang

    2012-12-06

    A reliable and simple test for the detection of malaria parasite is crucial in providing effective treatment and therapeutic follow-up, especially in malaria elimination programmes. A comparison of four methods, including nested polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) were used for the malaria diagnosis and treatment follow-up in São Tomé and Príncipe, during a successful pre-elimination campaign. During the period September to November 2009, blood samples from 128 children (five to 14 years old) with temperature ≥38°C (tympanic) in the District of Agua Grande were examined using four different methods, i.e., histidine-rich protein 2 (HRP-2) based rapid diagnostic tests (HRP-2-RDTs), optical microscopy, nested PCR, and LAMP. First-line treatment with artesunate-amodiaquine was given for uncomplicated malaria and intravenous quinine was given for complicated malaria. Children with persistent positivity for malaria by microscopy, or either by nested PCR, or by LAMP on day 7 were given second-line treatment with artemether-lumefantrine. Treatment follow-up was made weekly, for up to four weeks. On day 0, positive results for HRP-2-RDTs, microscopy, nested PCR, and LAMP, were 68(53%), 47(37%), 64(50%), and 65(51%), respectively. When nested PCR was used as a reference standard, only LAMP was comparable; both HRP-2-RDTs and microscopy had moderate sensitivity; HRP-2-RDTs had poor positive predictive value (PPV) and a moderate negative predictive value (NPV) for the treatment follow-up. Seventy-one children with uncomplicated malaria and eight children with complicated falciparum malaria were diagnosed based on at least one positive result from the four tests as well as clinical criteria. Twelve of the 79 children receiving first-line treatment had positive results by nested PCR on day 7 (nested PCR-corrected day 7 cure rate was 85%). After the second-line treatment, nested PCR/LAMP-corrected day 28 cure rate was 83% for these 12 children. HRP-2-RDTs have similar sensitivity as microscopy but less specificity. However, as compared to nested PCR, the poor sensitivity of HRP-2-RDTs indicates that low parasitaemia may not be detected after treatment, as well as the low specificity of HRP-2-RDTs indicates it cannot be applied for treatment follow-up. LAMP has similar sensitivity and specificity to nested PCR. With high PPV and NPV, LAMP is simpler and faster as compared to nested PCR with the advantage of detecting low parasitaemia becoming a potential point-of-care test for treatment follow-up.

  15. Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in São Tomé

    PubMed Central

    2012-01-01

    Background A reliable and simple test for the detection of malaria parasite is crucial in providing effective treatment and therapeutic follow-up, especially in malaria elimination programmes. A comparison of four methods, including nested polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) were used for the malaria diagnosis and treatment follow-up in São Tomé and Príncipe, during a successful pre-elimination campaign. Method During the period September to November 2009, blood samples from 128 children (five to 14 years old) with temperature ≥38°C (tympanic) in the District of Agua Grande were examined using four different methods, i.e., histidine-rich protein 2 (HRP-2) based rapid diagnostic tests (HRP-2-RDTs), optical microscopy, nested PCR, and LAMP. First-line treatment with artesunate-amodiaquine was given for uncomplicated malaria and intravenous quinine was given for complicated malaria. Children with persistent positivity for malaria by microscopy, or either by nested PCR, or by LAMP on day 7 were given second-line treatment with artemether-lumefantrine. Treatment follow-up was made weekly, for up to four weeks. Results On day 0, positive results for HRP-2-RDTs, microscopy, nested PCR, and LAMP, were 68(53%), 47(37%), 64(50%), and 65(51%), respectively. When nested PCR was used as a reference standard, only LAMP was comparable; both HRP-2-RDTs and microscopy had moderate sensitivity; HRP-2-RDTs had poor positive predictive value (PPV) and a moderate negative predictive value (NPV) for the treatment follow-up. Seventy-one children with uncomplicated malaria and eight children with complicated falciparum malaria were diagnosed based on at least one positive result from the four tests as well as clinical criteria. Twelve of the 79 children receiving first-line treatment had positive results by nested PCR on day 7 (nested PCR-corrected day 7 cure rate was 85%). After the second-line treatment, nested PCR/LAMP-corrected day 28 cure rate was 83% for these 12 children. Conclusions HRP-2-RDTs have similar sensitivity as microscopy but less specificity. However, as compared to nested PCR, the poor sensitivity of HRP-2-RDTs indicates that low parasitaemia may not be detected after treatment, as well as the low specificity of HRP-2-RDTs indicates it cannot be applied for treatment follow-up. LAMP has similar sensitivity and specificity to nested PCR. With high PPV and NPV, LAMP is simpler and faster as compared to nested PCR with the advantage of detecting low parasitaemia becoming a potential point-of-care test for treatment follow-up. PMID:23217163

  16. High-resolution angle-resolved photoemission study of electronic structure and charge-density wave formation in HoTe3

    NASA Astrophysics Data System (ADS)

    Liu, Guodong; Wang, Chenlu; Zhang, Yan; Hu, Bingfeng; Mou, Daixiang; Yu, Li; Zhao, Lin; Zhou, Xingjiang; Wang, Nanlin; Chen, Chuangtian; Xu, Zuyan

    We performed high-resolution angle-resolved photoemission spectroscopy (ARPES) measurement on high quality crystal of HoTe3, an intriguing quasi-two-dimensional rare-earth-element tritelluride charge-density-wave (CDW) compound. The main features of the electronic structure in this compound are established by employing a quasi-CW laser (7eV) and a helium discharging lamp (21.22 eV) as excitation light sources. It reveals many bands back folded according to the CDW periodicity and two incommensurate CDW gaps created by perpendicular Fermi surface (FS) nesting vectors. A large gap is found to open in well nested regions of the Fermi surface sheets, whereas other Fermi surface sections with poor nesting remain ungapped. In particular, some peculiar features are identified by using our ultra-high resolution and bulk sensitive laser-ARPES.

  17. Human papillomavirus detection using the Abbott RealTime high-risk HPV tests compared with conventional nested PCR coupled to high-throughput sequencing of amplification products in cervical smear specimens from a Gabonese female population.

    PubMed

    Moussavou-Boundzanga, Pamela; Koumakpayi, Ismaël Hervé; Labouba, Ingrid; Leroy, Eric M; Belembaogo, Ernest; Berthet, Nicolas

    2017-12-21

    Cervical cancer is the fourth most common malignancy in women worldwide. However, screening with human papillomavirus (HPV) molecular tests holds promise for reducing cervical cancer incidence and mortality in low- and middle-income countries. The performance of the Abbott RealTime High-Risk HPV test (AbRT) was evaluated in 83 cervical smear specimens and compared with a conventional nested PCR coupled to high-throughput sequencing (HTS) to identify the amplicons. The AbRT assay detected at least one HPV genotype in 44.57% of women regardless of the grade of cervical abnormalities. Except for one case, good concordance was observed for the genotypes detected with the AbRT assay in the high-risk HPV category determined with HTS of the amplicon generated by conventional nested PCR. The AbRT test is an easy and reliable molecular tool and was as sensitive as conventional nested PCR in cervical smear specimens for detection HPVs associated with high-grade lesions. Moreover, sequencing amplicons using an HTS approach effectively identified the genotype of the hrHPV identified with the AbRT test.

  18. Detection of Mycobacterium tuberculosis in extrapulmonary biopsy samples using PCR targeting IS6110, rpoB, and nested-rpoB PCR Cloning

    PubMed Central

    Meghdadi, Hossein; Khosravi, Azar D.; Ghadiri, Ata A.; Sina, Amir H.; Alami, Ameneh

    2015-01-01

    Present study was aimed to examine the diagnostic utility of polymerase chain reaction (PCR) and nested PCR techniques for the detection of Mycobacterium tuberculosis (MTB) DNA in samples from patients with extra pulmonary tuberculosis (EPTB). In total 80 formalin-fixed, paraffin-embedded (FFPE) samples comprising 70 samples with definite diagnosis of EPTB and 10 samples from known non- EPTB on the basis of histopathology examination, were included in the study. PCR amplification targeting IS6110, rpoB gene and nested PCR targeting the rpoB gene were performed on the extracted DNAs from 80 FFPE samples. The strong positive samples were directly sequenced. For negative samples and those with weak band in nested-rpoB PCR, TA cloning was performed by cloning the products into the plasmid vector with subsequent sequencing. The 95% confidence intervals (CI) for the estimates of sensitivity and specificity were calculated for each method. Fourteen (20%), 34 (48.6%), and 60 (85.7%) of the 70 positive samples confirmed by histopathology, were positive by rpoB-PCR, IS6110-PCR, and nested-rpoB PCR, respectively. By performing TA cloning on samples that yielded weak (n = 8) or negative results (n = 10) in the PCR methods, we were able to improve their quality for later sequencing. All samples with weak band and 7 out of 10 negative samples, showed strong positive results after cloning. So nested-rpoB PCR cloning revealed positivity in 67 out of 70 confirmed samples (95.7%). The sensitivity of these combination methods was calculated as 95.7% in comparison with histopathology examination. The CI for sensitivity of the PCR methods were calculated as 11.39–31.27% for rpoB-PCR, 36.44–60.83% for IS6110- PCR, 75.29–92.93% for nested-rpoB PCR, and 87.98–99.11% for nested-rpoB PCR cloning. The 10 true EPTB negative samples by histopathology, were negative by all tested methods including cloning and were used to calculate the specificity of the applied methods. The CI for 100% specificity of each PCR method were calculated as 69.15–100%. Our results indicated that nested-rpoB PCR combined with TA cloning and sequencing is a preferred method for the detection of MTB DNA in EPTB samples with high sensitivity and specificity which confirm the histopathology results. PMID:26191059

  19. Detection of Mycobacterium tuberculosis in extrapulmonary biopsy samples using PCR targeting IS6110, rpoB, and nested-rpoB PCR Cloning.

    PubMed

    Meghdadi, Hossein; Khosravi, Azar D; Ghadiri, Ata A; Sina, Amir H; Alami, Ameneh

    2015-01-01

    Present study was aimed to examine the diagnostic utility of polymerase chain reaction (PCR) and nested PCR techniques for the detection of Mycobacterium tuberculosis (MTB) DNA in samples from patients with extra pulmonary tuberculosis (EPTB). In total 80 formalin-fixed, paraffin-embedded (FFPE) samples comprising 70 samples with definite diagnosis of EPTB and 10 samples from known non- EPTB on the basis of histopathology examination, were included in the study. PCR amplification targeting IS6110, rpoB gene and nested PCR targeting the rpoB gene were performed on the extracted DNAs from 80 FFPE samples. The strong positive samples were directly sequenced. For negative samples and those with weak band in nested-rpoB PCR, TA cloning was performed by cloning the products into the plasmid vector with subsequent sequencing. The 95% confidence intervals (CI) for the estimates of sensitivity and specificity were calculated for each method. Fourteen (20%), 34 (48.6%), and 60 (85.7%) of the 70 positive samples confirmed by histopathology, were positive by rpoB-PCR, IS6110-PCR, and nested-rpoB PCR, respectively. By performing TA cloning on samples that yielded weak (n = 8) or negative results (n = 10) in the PCR methods, we were able to improve their quality for later sequencing. All samples with weak band and 7 out of 10 negative samples, showed strong positive results after cloning. So nested-rpoB PCR cloning revealed positivity in 67 out of 70 confirmed samples (95.7%). The sensitivity of these combination methods was calculated as 95.7% in comparison with histopathology examination. The CI for sensitivity of the PCR methods were calculated as 11.39-31.27% for rpoB-PCR, 36.44-60.83% for IS6110- PCR, 75.29-92.93% for nested-rpoB PCR, and 87.98-99.11% for nested-rpoB PCR cloning. The 10 true EPTB negative samples by histopathology, were negative by all tested methods including cloning and were used to calculate the specificity of the applied methods. The CI for 100% specificity of each PCR method were calculated as 69.15-100%. Our results indicated that nested-rpoB PCR combined with TA cloning and sequencing is a preferred method for the detection of MTB DNA in EPTB samples with high sensitivity and specificity which confirm the histopathology results.

  20. Molecular and clinical analyses of Helicobacter pylori colonization in inflamed dental pulp.

    PubMed

    Nomura, Ryota; Ogaya, Yuko; Matayoshi, Saaya; Morita, Yumiko; Nakano, Kazuhiko

    2018-04-16

    Recently, dental pulp has been considered a possible source of infection of Helicobacter pylori (H. pylori) in children. We previously developed a novel PCR system for H. pylori detection with high specificity and sensitivity using primer sets constructed based on the complete genome information for 48 H. pylori strains. This PCR system showed high sensitivity with a detection limit of 1-10 cells when serial dilutions of H. pylori genomic DNA were used as templates. However, the detection limit was lower (10 2 -10 3 cells) when H. pylori bacterial DNA was detected from inflamed pulp specimens. Thus, we further refined the system using a nested PCR method, which was much more sensitive than the previous single PCR method. In addition, we examined the distribution and virulence of H. pylori in inflamed pulp tissue. Nested PCR system was constructed using primer sets designed from the complete genome information of 48 H. pylori strains. The detection limit of the nested PCR system was 1-10 cells using both H. pylori genomic DNA and bacterial DNA isolated from inflamed pulp specimens. Next, distribution of H. pylori was examined using 131 inflamed pulp specimens with the nested PCR system. In addition, association between the detection of H. pylori and clinical information regarding endodontic-infected teeth were investigated. Furthermore, adhesion property of H. pylori strains to human dental fibroblast cells was examined. H. pylori was present in 38.9% of inflamed pulp specimens using the nested PCR system. H. pylori was shown to be predominantly detected in primary teeth rather than permanent teeth. In addition, samplings of the inflamed pulp were performed twice from the same teeth at 1- or 2-week intervals, which revealed that H. pylori was detected in most specimens in both samplings. Furthermore, H. pylori strains showed adhesion property to human dental fibroblast cells. Our results suggest that H. pylori colonizes inflamed pulp in approximately 40% of all cases through adhesion to human dental fibroblast cells.

  1. Population dynamics of long-tailed ducks breeding on the Yukon-Kuskokwim Delta, Alaska

    USGS Publications Warehouse

    Schamber, Jason L.; Flint, Paul L.; Grand, J. Barry; Wilson, Heather M.; Morse, Julie A.

    2009-01-01

    Population estimates for long-tailed ducks in North America have declined by nearly 50% over the past 30 years. Life history and population dynamics of this species are difficult to ascertain, because the birds nest at low densities across a broad range of habitat types. Between 1991 and 2004, we collected information on productivity and survival of long-tailed ducks at three locations on the Yukon-Kuskokwim Delta. Clutch size averaged 7.1 eggs, and nesting success averaged 30%. Duckling survival to 30 days old averaged 10% but was highly variable among years, ranging from 0% to 25%. Apparent annual survival of adult females based on mark-recapture of nesting females was estimated at 74%. We combined these estimates of survival and productivity into a matrix-based population model, which predicted an annual population decline of 19%. Elasticities indicated that population growth rate (λ) was most sensitive to changes in adult female survival. Further, the relatively high sensitivity of λ to duckling survival suggests that low duckling survival may be a bottleneck to productivity in some years. These data represent the first attempt to synthesize a population model for this species. Although our analyses were hampered by the small sample sizes inherent in studying a dispersed nesting species, our model provides a basis for management actions and can be enhanced as additional data become available.

  2. Challenging loop-mediated isothermal amplification (LAMP) technique for molecular detection of Toxoplasma gondii.

    PubMed

    Fallahi, Shirzad; Mazar, Zahra Arab; Ghasemian, Mehrdad; Haghighi, Ali

    2015-05-01

    To compare analytical sensitivity and specificity of a newly described DNA amplification technique, LAMP and nested PCR assay targeting the RE and B1 genes for the detection of Toxoplasma gondii (T. gondii) DNA. The analytical sensitivity of LAMP and nested-PCR was obtained against10-fold serial dilutions of T. gondii DNA ranging from 1 ng to 0.01 fg. DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays. After testing LAMP and nested-PCR in duplicate, the detection limit of RE-LAMP, B1-LAMP, RE-nested PCR and B1-nested PCR assays was one fg, 100 fg, 1 pg and 10 pg of T. gondii DNA respectively. All the LAMP assays and nested PCRs were 100% specific. The RE-LAMP assay revealed the most sensitivity for the detection of T. gondii DNA. The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T. gondii. Furthermore, these findings indicate that primers based on the RE are more suitable than those based on the B1 gene. However, the B1-LAMP assay has potential as a diagnostic tool for detection of T. gondii. Copyright © 2015 Hainan Medical College. Production and hosting by Elsevier B.V. All rights reserved.

  3. Whole-Blood Nested-PCR Amplification of M. leprae-Specific DNA for Early Diagnosis of Leprosy

    PubMed Central

    Wen, Yan; Xing, Yan; Yuan, Lian-Chao; Liu, Jian; Zhang, Ying; Li, Huan-Ying

    2013-01-01

    We evaluated the sensitivity and specificity of a nested-polymerase chain reaction (PCR) method for detection of Mycobacterium leprae DNA from whole blood. Whole-blood specimens were subjected to nested-PCR amplification of M. leprae repeat DNA sequences in 49 multibacillary (MB) and 30 paucibacillary (PB) leprosy patients, 96 household contacts (HHCs), 18 tuberculosis (TB) patients, and 35 normal healthy individuals. M. leprae DNA was detected in 95.92% (47/49) of MB, 70% (21/30) of PB, and 6.25% (6/96) of HHC, but it was not detected in 18 TB or 35 normal controls. The sensitivities of the anti-bovine serum albumin (ND-O-BSA) immunoglobulin M (IgM) and antifusion protein of ML0405-ML2331 IgG for MB were 97.96% and 89.8%, and these values for PB were 70% and 53.33%. However, the ND-O-BSA enzyme-linked immunosorbent assay (ELISA) had lower specificity, with relatively high false-positive results for TB patients (16.67%) and normal healthy controls (10%). Based on these promising findings, we propose the use of nested PCR of whole-blood samples along with ELISA test for early detection of leprosy cases. PMID:23478578

  4. Early diagnosis of typhoid fever by nested PCR for flagellin gene of Salmonella enterica serotype Typhi.

    PubMed

    Khan, S; Harish, B N; Menezes, G A; Acharya, N S; Parija, S C

    2012-11-01

    Typhoid fever caused by Salmonella Typhi continues to be a major health problem in spite of the use of antibiotics and the development of newer antibacterial drugs. Inability to make an early laboratory diagnosis and resort to empirical therapy, often lead to increased morbidity and mortality in cases of typhoid fever. This study was aimed to optimize a nested PCR for early diagnosis of typhoid fever and using it as a diagnostic tool in culture negative cases of suspected typhoid fever. Eighty patients with clinical diagnosis of typhoid fever and 40 controls were included in the study. The blood samples collected were subjected to culture, Widal and nested PCR targeting the flagellin gene of S. Typhi. The sensitivity of PCR on blood was found to be 100 per cent whereas the specificity was 76.9 per cent. The positive predictive value (PPV) of PCR was calculated to be 76.9 per cent with an accuracy of 86 per cent. None of the 40 control samples gave a positive PCR. Due to its high sensitivity and specificity nested PCR can be used as a useful tool to diagnose clinically suspected, culture negative cases of typhoid fever.

  5. Differentiation of Mycobacterium tuberculosis complex from non-tubercular mycobacteria by nested multiplex PCR targeting IS6110, MTP40 and 32kD alpha antigen encoding gene fragments.

    PubMed

    Sinha, Pallavi; Gupta, Anamika; Prakash, Pradyot; Anupurba, Shampa; Tripathi, Rajneesh; Srivastava, G N

    2016-03-12

    Control of the global burden of tuberculosis is obstructed due to lack of simple, rapid and cost effective diagnostic techniques that can be used in resource poor-settings. To facilitate the early diagnosis of TB directly from clinical specimens, we have standardized and validated the use of nested multiplex PCR, targeting gene fragments IS6110, MTP40 and 32kD α-antigen encoding genes specific for Mycobacterium tuberculosis complex and non-tubercular mycobacteria (NTM), in comparison to smear microscopy, solid culture and single step multiplex PCR. The results were evaluated in comparison to a composite reference standard (CRS) comprising of microbiological results (smear and culture), clinical, radiological and cytopathological findings, clinical treatment and response to anti-tubercular therapy. The nested multiplex PCR (nMPCR) assay was evaluated to test its utility in 600 (535 pulmonary and 65 extra-pulmonary specimens) clinically suspected TB cases. All specimens were processed for smear, culture, single step multiplex PCR and nested multiplex PCR testing. Out of 535 screened pulmonary and 65 extra-pulmonary specimens, 329 (61.5%) and 19 (29.2%) cases were culture positive for M. tuberculosis. Based on CRS, 450 patients had "clinical TB" (definitive-TB, probable-TB and possible-TB). Remaining 150 were confirmed "non-TB" cases. For culture, the sensitivity was low, 79.3% for pulmonary and 54.3% for extra-pulmonary cases. The sensitivity and specificity results for nMPCR test were evaluated taken composite reference standard as a gold standard. The sensitivity of the nMPCR assay was 97.1% for pulmonary and 91.4% for extra-pulmonary TB cases with specificity of 100% and 93.3% respectively. Nested multiplex PCR using three gene primers is a rapid, reliable and highly sensitive and specific diagnostic technique for the detection and differentiation of M. tuberculosis complex from NTM genome and will be useful in diagnosing paucibacillary samples. Nested multiplex PCR assay was found to be better than single step multiplex PCR for assessing the diagnosis of TB.

  6. Comparison of Nested Polymerase Chain Reaction and Real-Time Polymerase Chain Reaction with Parasitological Methods for Detection of Strongyloides stercoralis in Human Fecal Samples

    PubMed Central

    Sharifdini, Meysam; Mirhendi, Hossein; Ashrafi, Keyhan; Hosseini, Mostafa; Mohebali, Mehdi; Khodadadi, Hossein; Kia, Eshrat Beigom

    2015-01-01

    This study was performed to evaluate nested polymerase chain reaction (PCR) and real-time PCR methods for detection of Strongyloides stercoralis in fecal samples compared with parasitological methods. A total of 466 stool samples were examined by conventional parasitological methods (formalin ether concentration [FEC] and agar plate culture [APC]). DNA was extracted using an in-house method, and mitochondrial cytochrome c oxidase subunit 1 and 18S ribosomal genes were amplified by nested PCR and real-time PCR, respectively. Among 466 samples, 12.7% and 18.2% were found infected with S. stercoralis by FEC and APC, respectively. DNA of S. stercoralis was detected in 18.9% and 25.1% of samples by real-time PCR and nested PCR, respectively. Considering parasitological methods as the diagnostic gold standard, the sensitivity and specificity of nested PCR were 100% and 91.6%, respectively, and that of real-time PCR were 84.7% and 95.8%, respectively. However, considering sequence analyzes of the selected nested PCR products, the specificity of nested PCR is increased. In general, molecular methods were superior to parasitological methods. They were more sensitive and more reliable in detection of S. stercoralis in comparison with parasitological methods. Between the two molecular methods, the sensitivity of nested PCR was higher than real-time PCR. PMID:26350449

  7. Osprey: worldwide sentinel species for assessing and monitoring environmental contamination in rivers, lakes, reservoirs, and estuaries.

    PubMed

    Grove, Robert A; Henny, Charles J; Kaiser, James L

    2009-01-01

    In the United States, many fish and wildlife species have been used nationwide to monitor environmental contaminant exposure and effects, including carcasses of the bald eagle (Haliaeetus leucocephalus), the only top avian predator regularly used in the past. Unfortunately, bald eagles are sensitive to investigator intrusion at the nest. Thus, the osprey (Pandion haliaetus) is evaluated as a potential sentinel species for aquatic ecosystems. Several characteristics support the choice of the osprey as a sentinel species, including: (1) fish-eating diet atop the aquatic food web, (2) long-lived with strong nest fidelity, (3) adapts to human landscapes (potentially the most contaminated), (4) tolerates short-term nest disturbance, (5) nests spatially distributed at regular intervals, (6) highly visible nests easily located for study, (7) ability to accumulate most, if not all, lipophilic contaminants, (8) known sensitivity to many contaminants, and (9) nearly a worldwide distribution. These osprey traits have been instrumental in successfully using the species to understand population distribution, abundance, and changes over time; the effects of various contaminants on reproductive success; how contaminants in prey (fish on biomass basis) contribute to egg concentrations (i.e., biomagnification factors); and spatial residue patterns. Data summarized include nesting population surveys, detailed nesting studies, and chemical analyses of osprey egg, organ, blood, and feather samples for contaminants that bioaccumulate and/or biomagnify in aquatic food webs; and biochemical evaluations of blood and various organs. Studies in the United States, Canada, Mexico, Europe, and elsewhere have shown the osprey to be a useful sentinel species for monitoring selected environmental contaminants, including some emerging contaminants in lakes, reservoirs, rivers, and estuaries.

  8. SENSITIVITY OF NEST SUCCESS, YOUNG FLEDGED, AND PROBABILITY OF RENESTING TO SEASONAL FECUNDITY IN MULTI-BROODED SPECIES

    EPA Science Inventory

    A considerable number of avian species can produce multiple broods within a season. Seasonal fecundity in these species can vary by changes in the number of young fledged per nest, the probability of a successful nest, and the probability of initiating additional nests (e.g., re...

  9. Development of a novel single tube nested PCR for enhanced detection of cytomegalovirus DNA from dried blood spots.

    PubMed

    Atkinson, C; Emery, V C; Griffiths, P D

    2014-02-01

    Newborn screening for congenital cytomegalovirus (CCMV) using dried blood spots (DBS) has been proposed because many developed countries have DBS screening programmes in place for other diseases. The aim of this study was to develop a rapid, single tube nested polymerase chain reaction (PCR) method for enhanced detection of CMV from DBS compared to existing (single target) real time PCRs. The new method was compared with existing real time PCRs for sensitivity and specificity. Overall sensitivity of the single target PCR assays in both asymptomatic and symptomatic infants with laboratory confirmed congenital CMV was 69% (CMV PCR or culture positive before day 21 of life). In contrast, the single tube nested assay had an increased sensitivity of 81% with100% specificity. Overall the assay detected CMV from a DBS equivalent to an original blood sample which contained 500IU/ml. In conclusion this single tube nested methodology allows simultaneous amplification and detection of CMV DNA in 1.5h removing the associated contamination risk of a two step nested PCR. Owing to its increased sensitivity, it has the potential to be used as a screening assay and ultimately allow early identification and intervention for children with congenital CMV. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Comparison of Microscopy, Nested-PCR, and Real-Time-PCR Assays Using High-Throughput Screening of Pooled Samples for Diagnosis of Malaria in Asymptomatic Carriers from Areas of Endemicity in Myanmar

    PubMed Central

    Wang, Bo; Han, Soe-Soe; Cho, Cho; Han, Jin-Hee; Cheng, Yang; Lee, Seong-Kyun; Galappaththy, Gawrie N. L; Thimasarn, Krongthong; Soe, Myat Thu; Oo, Htet Wai; Kyaw, Myat Phone

    2014-01-01

    Asymptomatic infection is an important obstacle for controlling disease in countries where malaria is endemic. Because asymptomatic carriers do not seek treatment for their infections, they can have high levels of gametocytes and constitute a reservoir available for new infection. We employed a sample pooling/PCR-based molecular detection strategy for screening malaria infection in residents from areas of Myanmar where malaria is endemic. Blood samples (n = 1,552) were collected from residents in three areas of malaria endemicity (Kayin State, Bago, and Tanintharyi regions) of Myanmar. Two nested PCR and real-time PCR assays showed that asymptomatic infection was detected in about 1.0% to 9.4% of residents from the surveyed areas. The sensitivities of the two nested PCR and real-time PCR techniques were higher than that of microscopy examination (sensitivity, 100% versus 26.4%; kappa values, 0.2 to 0.5). Among the three regions, parasite-positive samples were highly detected in subjects from the Bago and Tanintharyi regions. Active surveillance of residents from regions of intense malaria transmission would reduce the risk of morbidity and mitigate transmission to the population in these areas of endemicity. Our data demonstrate that PCR-based molecular techniques are more efficient than microscopy for nationwide surveillance of malaria in countries where malaria is endemic. PMID:24648557

  11. Comparison of Two Grid Refinement Approaches for High Resolution Regional Climate Modeling: MPAS vs WRF

    NASA Astrophysics Data System (ADS)

    Leung, L.; Hagos, S. M.; Rauscher, S.; Ringler, T.

    2012-12-01

    This study compares two grid refinement approaches using global variable resolution model and nesting for high-resolution regional climate modeling. The global variable resolution model, Model for Prediction Across Scales (MPAS), and the limited area model, Weather Research and Forecasting (WRF) model, are compared in an idealized aqua-planet context with a focus on the spatial and temporal characteristics of tropical precipitation simulated by the models using the same physics package from the Community Atmosphere Model (CAM4). For MPAS, simulations have been performed with a quasi-uniform resolution global domain at coarse (1 degree) and high (0.25 degree) resolution, and a variable resolution domain with a high-resolution region at 0.25 degree configured inside a coarse resolution global domain at 1 degree resolution. Similarly, WRF has been configured to run on a coarse (1 degree) and high (0.25 degree) resolution tropical channel domain as well as a nested domain with a high-resolution region at 0.25 degree nested two-way inside the coarse resolution (1 degree) tropical channel. The variable resolution or nested simulations are compared against the high-resolution simulations that serve as virtual reality. Both MPAS and WRF simulate 20-day Kelvin waves propagating through the high-resolution domains fairly unaffected by the change in resolution. In addition, both models respond to increased resolution with enhanced precipitation. Grid refinement induces zonal asymmetry in precipitation (heating), accompanied by zonal anomalous Walker like circulations and standing Rossby wave signals. However, there are important differences between the anomalous patterns in MPAS and WRF due to differences in the grid refinement approaches and sensitivity of model physics to grid resolution. This study highlights the need for "scale aware" parameterizations in variable resolution and nested regional models.

  12. [Optimized application of nested PCR method for detection of malaria].

    PubMed

    Yao-Guang, Z; Li, J; Zhen-Yu, W; Li, C

    2017-04-28

    Objective To optimize the application of the nested PCR method for the detection of malaria according to the working practice, so as to improve the efficiency of malaria detection. Methods Premixing solution of PCR, internal primers for further amplification and new designed primers that aimed at two Plasmodium ovale subspecies were employed to optimize the reaction system, reaction condition and specific primers of P . ovale on basis of routine nested PCR. Then the specificity and the sensitivity of the optimized method were analyzed. The positive blood samples and examination samples of malaria were detected by the routine nested PCR and the optimized method simultaneously, and the detection results were compared and analyzed. Results The optimized method showed good specificity, and its sensitivity could reach the pg to fg level. The two methods were used to detect the same positive malarial blood samples simultaneously, the results indicated that the PCR products of the two methods had no significant difference, but the non-specific amplification reduced obviously and the detection rates of P . ovale subspecies improved, as well as the total specificity also increased through the use of the optimized method. The actual detection results of 111 cases of malarial blood samples showed that the sensitivity and specificity of the routine nested PCR were 94.57% and 86.96%, respectively, and those of the optimized method were both 93.48%, and there was no statistically significant difference between the two methods in the sensitivity ( P > 0.05), but there was a statistically significant difference between the two methods in the specificity ( P < 0.05). Conclusion The optimized PCR can improve the specificity without reducing the sensitivity on the basis of the routine nested PCR, it also can save the cost and increase the efficiency of malaria detection as less experiment links.

  13. Sensitivity studies of high-resolution RegCM3 simulations of precipitation over the European Alps: the effect of lateral boundary conditions and domain size

    NASA Astrophysics Data System (ADS)

    Nadeem, Imran; Formayer, Herbert

    2016-11-01

    A suite of high-resolution (10 km) simulations were performed with the International Centre for Theoretical Physics (ICTP) Regional Climate Model (RegCM3) to study the effect of various lateral boundary conditions (LBCs), domain size, and intermediate domains on simulated precipitation over the Great Alpine Region. The boundary conditions used were ECMWF ERA-Interim Reanalysis with grid spacing 0.75∘, the ECMWF ERA-40 Reanalysis with grid spacing 1.125 and 2.5∘, and finally the 2.5∘ NCEP/DOE AMIP-II Reanalysis. The model was run in one-way nesting mode with direct nesting of the high-resolution RCM (horizontal grid spacing Δx = 10 km) with driving reanalysis, with one intermediate resolution nest (Δx = 30 km) between high-resolution RCM and reanalysis forcings, and also with two intermediate resolution nests (Δx = 90 km and Δx = 30 km) for simulations forced with LBC of resolution 2.5∘. Additionally, the impact of domain size was investigated. The results of multiple simulations were evaluated using different analysis techniques, e.g., Taylor diagram and a newly defined useful statistical parameter, called Skill-Score, for evaluation of daily precipitation simulated by the model. It has been found that domain size has the major impact on the results, while different resolution and versions of LBCs, e.g., 1.125∘ ERA40 and 0.7∘ ERA-Interim, do not produce significantly different results. It is also noticed that direct nesting with reasonable domain size, seems to be the most adequate method for reproducing precipitation over complex terrain, while introducing intermediate resolution nests seems to deteriorate the results.

  14. The role of pleural fluid MAGE RT-nested PCR in the diagnosis of malignant pleural effusion.

    PubMed

    Jeon, Eun Ju; Park, Hye Kyeong; Jeon, Kyeongman; Koh, Won-Jung; Suh, Gee Young; Chung, Man Pyo; Kim, Hojoong; Kwon, O Jung; Ki, Chang-Seok; Kim, Jong-Won; Shim, Young Mog; Um, Sang-Won

    2012-11-01

      Melanoma antigen (MAGE) genes are expressed in tumor cells, the testis and the placenta. The purpose of this prospective study was to investigate the sensitivity, specificity, and accuracy of the carcinoembryonic antigen (CEA), MAGE reverse transcriptase-nested polymerase chain reaction (RT-nested PCR), and cytology of pleural fluid in the diagnosis of malignant pleural effusion.   Patients in whom unilateral pleural effusion was identified on chest radiography from January to December 2009 were included in the study. MAGE genes were analyzed by RT-nested PCR using MAGE A1-6 common primers.   Of 81 enrolled patients, 46 were diagnosed as malignant pleural effusion, and 24 were diagnosed as benign pleural effusion. The diagnoses of 11 patients were not confirmed in this study. The diagnostic sensitivity, specificity, and accuracy of MAGE RT-nested PCR were 61.4%, 95.7%, and 73.1%, respectively. The diagnostic sensitivities of cytology and CEA (>5 ng/mL) were 61.4% and 75.0%, respectively. Among 17 patients with negative cytology who had malignant pleural effusion, 12 and 10 patients were positive for CEA (>5.0 ng/mL) and MAGE RT-nested PCR, respectively. However, of five patients with malignant pleural effusion that was not recognized by cytology and CEA, MAGE RT-nested PCR correctly predicted a malignant etiology in only one additional patient (20%).   MAGE RT-nested PCR seems to add little on the combination of conventional methods in the diagnosis of malignant effusion. © 2012 Tianjin Lung Cancer Institute and Wiley Publishing Asia Pty. Ltd.

  15. Validation and clinical application of a molecular method for the identification of Cryptococcus neoformans/Cryptococcus gattii complex DNA in human clinical specimens.

    PubMed

    Rivera, Vanessa; Gaviria, Marcela; Muñoz-Cadavid, Cesar; Cano, Luz; Naranjo, Tonny

    2015-01-01

    The diagnosis of cryptococcosis is usually performed based on cultures of tissue or body fluids and isolation of the fungus, but this method may require several days. Direct microscopic examination, although rapid, is relatively insensitive. Biochemical and immunodiagnostic rapid tests are also used. However, all of these methods have limitations that may hinder final diagnosis. The increasing incidence of fungal infections has focused attention on tools for rapid and accurate diagnosis using molecular biological techniques. Currently, PCR-based methods, particularly nested, multiplex and real-time PCR, provide both high sensitivity and specificity. In the present study, we evaluated a nested PCR targeting the gene encoding the ITS-1 and ITS-2 regions of rDNA in samples from a cohort of patients diagnosed with cryptococcosis. The results showed that in our hands, this Cryptococcus nested PCR assay has 100% specificity and 100% sensitivity and was able to detect until 2 femtograms of Cryptococcus DNA. Copyright © 2015 Elsevier Editora Ltda. All rights reserved.

  16. Improved clonality detection in B-cell lymphoma using a semi-nested modification of the BIOMED-2 PCR assay for IGH rearrangement: A paraffin-embedded tissue study.

    PubMed

    Sakamoto, Yuma; Masaki, Ayako; Aoyama, Satsuki; Han, Shusen; Saida, Kosuke; Fujii, Kana; Takino, Hisashi; Murase, Takayuki; Iida, Shinsuke; Inagaki, Hiroshi

    2017-09-01

    The BIOMED-2 PCR protocol for targeting the IGH gene is widely employed for detecting clonality in B-cell malignancies. Unfortunately, the detection of clonality with this method is not very sensitive when paraffin sections are used as a DNA source. To increase the sensitivity, we devised a semi-nested modification of a JH consensus primer. The clonality detection rates of three assays were compared: the standard BIOMED-2, BIOMED-2 assay followed by BIOMED-2 re-amplification, and BIOMED-2 assay followed by semi-nested BIOMED-2. We tested more than 100 cases using paraffin-embedded tissues of various B-cell lymphomas, and found that the clonality detection rates with the above three assays were 63.9%, 79.6%, and 88.0%, respectively. While BIOMED-2 re-amplification was significantly more sensitive than the standard BIOMED-2, the semi-nested BIOMED-2 was significantly more sensitive than both the standard BIOMED-2 and BIOMED-2 re-amplification. An increase in sensitivity was observed in all lymphoma subtypes examined. In conclusion, tumor clonality may be detected in nearly 90% of B-cell lymphoma cases with semi-nested BIOMED-2. This ancillary assay may be useful when the standard BIOMED-2 fails to detect clonality in histopathologically suspected B-cell lymphomas. © 2017 Japanese Society of Pathology and John Wiley & Sons Australia, Ltd.

  17. Evaluation by latent class analysis of a magnetic capture based DNA extraction followed by real-time qPCR as a new diagnostic method for detection of Echinococcus multilocularis in definitive hosts.

    PubMed

    Maas, Miriam; van Roon, Annika; Dam-Deisz, Cecile; Opsteegh, Marieke; Massolo, Alessandro; Deksne, Gunita; Teunis, Peter; van der Giessen, Joke

    2016-10-30

    A new method, based on a magnetic capture based DNA extraction followed by qPCR, was developed for the detection of the zoonotic parasite Echinococcus multilocularis in definitive hosts. Latent class analysis was used to compare this new method with the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. In total, 60 red foxes and coyotes from three different locations were tested with both molecular methods and the sedimentation and counting technique (SCT) or intestinal scraping technique (IST). Though based on a limited number of samples, it could be established that the magnetic capture based DNA extraction followed by qPCR showed similar sensitivity and specificity as the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. All methods have a high specificity as shown by Bayesian latent class analysis. Both molecular assays have higher sensitivities than the combined SCT and IST, though the uncertainties in sensitivity estimates were wide for all assays tested. The magnetic capture based DNA extraction followed by qPCR has the advantage of not requiring hazardous chemicals like the phenol-chloroform DNA extraction followed by single tube nested PCR. This supports the replacement of the phenol-chloroform DNA extraction followed by single tube nested PCR by the magnetic capture based DNA extraction followed by qPCR for molecular detection of E. multilocularis in definitive hosts. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Vision on the high seas: spatial resolution and optical sensitivity in two procellariiform seabirds with different foraging strategies.

    PubMed

    Mitkus, Mindaugas; Nevitt, Gabrielle A; Danielsen, Johannis; Kelber, Almut

    2016-11-01

    Procellariiform or 'tubenosed' seabirds are challenged to find prey and orient over seemingly featureless oceans. Previous studies have found that life-history strategy (burrow versus surface nesting) was correlated to foraging strategy. Burrow nesters tended to track prey using dimethyl sulphide (DMS), a compound associated with phytoplankton, whereas surface-nesting species did not. Burrow nesters also tended to be smaller and more cryptic, whereas surface nesters were larger with contrasting plumage coloration. Together these results suggested that differences in life-history strategy might also be linked to differences in visual adaptations. Here, we used Leach's storm petrel, a DMS-responder, and northern fulmar, a non-responder, as model species to test this hypothesis on their sensory ecology. From the retinal ganglion cell density and photoreceptor dimensions, we determined that Leach's storm petrels have six times lower spatial resolution than the northern fulmars. However, the optical sensitivity of rod photoreceptors is similar between species. These results suggest that under similar atmospheric conditions, northern fulmars have six times the detection range for similarly sized objects. Both species have extended visual streaks with a central area of highest spatial resolution, but only the northern fulmar has a central fovea. The prediction that burrow-nesting DMS-responding procellariiforms should differ from non-responding species nesting in the open holds true for spatial resolution, but not for optical sensitivity. This result may reflect the fact that both species rely on olfaction for their nocturnal foraging activity, but northern fulmars might use vision more during daytime. © 2016. Published by The Company of Biologists Ltd.

  19. Detection of Mycoplasma hyopneumoniae by Air Sampling with a Nested PCR Assay

    PubMed Central

    Stärk, Katharina D. C.; Nicolet, Jacques; Frey, Joachim

    1998-01-01

    This article describes the first successful detection of airborne Mycoplasma hyopneumoniae under experimental and field conditions with a new nested PCR assay. Air was sampled with polyethersulfone membranes (pore size, 0.2 μm) mounted in filter holders. Filters were processed by dissolution and direct extraction of DNA for PCR analysis. For the PCR, two nested pairs of oligonucleotide primers were designed by using an M. hyopneumoniae-specific DNA sequence of a repeated gene segment. A nested PCR assay was developed and used to analyze samples collected in eight pig houses where respiratory problems had been common. Air was also sampled from a mycoplasma-free herd. The nested PCR was highly specific and 104 times as sensitive as a one-step PCR. Under field conditions, the sampling system was able to detect airborne M. hyopneumoniae on 80% of farms where acute respiratory disease was present. No airborne M. hyopneumoniae was detected on infected farms without acute cases. The chance of successful detection was increased if air was sampled at several locations within a room and at a lower air humidity. PMID:9464391

  20. Relation of water level and fish availability to wood stork reproduction in the southern Everglades, Florida

    USGS Publications Warehouse

    Kushlan, James A.; Ogden, John C.; Higer, Aaron L.

    1975-01-01

    The wood stork is a species of colonial wading bird in the Everglades that is most sensitive to changes in the availability of food. Previous studies have shown that the initiation and success of wood stork nesting depends on high densities of fish concentrated in ponds and other catchment basins during the dry season. The extreme dependence of the wood stork on the cyclic hydrologic regime of the southern Florida wetlands makes it an indicator of the well-being and ecological stability of the Everglades. The wood stork has declined in numbers over the last 25 years. One reason for the decline in wood stork population was the change in the hydrologic regimen of the Everglades which affected the feeding habitat and the food production. The fish on which the wood stork feeds increase in density during the dry season as water levels fall. In the Everglades marsh, densities were highest in front of the drying edge of surface water at a depth of about 0.3 m. Dry-season densities were greatest when a drought occurred the previous year. Historically wood stork nesting success was associated with high summer water levels, high rates of surface-water discharge and high rates of drying. Before the closure of the south side of Conservation Area 3 in 1962, years of successful and unsuccessful nesting were characterized by different patterns of drying. These patterns changed after 1962 and generally the predictability of successful nesting breaks down thereafter. Only two nesting years after 1962 were successful and in only one of these was the drying rate similar to years of successful nesting before 1962. Two other potentially successful years failed after 1962. This suggests that further changes in the hydrobiological relations occurred within the Everglades after 1962. Lack of successful nesting after 1962 can be attributed in large part to late colony formation and the interruption of nesting by winter rainfall. In this period (1962-72), colonies formed earlier in years of high early drying rates than in years of low early drying rates. Delay of colony formation is ultimately the result of inability to attain a suitable nutritional state since food supply is the primary factor in the initiation of nesting. Many of the complex food associations of the wood stork remain to be explained.

  1. High-Resolution Melting-Curve Analysis of obg Gene to Differentiate the Temperature-Sensitive Mycoplasma synoviae Vaccine Strain MS-H from Non-Temperature-Sensitive Strains

    PubMed Central

    Shahid, Muhammad A.; Markham, Philip F.; Marenda, Marc S.; Agnew-Crumpton, Rebecca; Noormohammadi, Amir H.

    2014-01-01

    Temperature-sensitive (ts +) vaccine strain MS-H is the only live attenuated M. synoviae vaccine commercially available for use in poultry. With increasing use of this vaccine to control M. synoviae infections, differentiation of MS-H from field M. synoviae strains and from rarely occurring non-temperature-sensitive (ts –) MS-H revertants has become important, especially in countries where local strains are indistinguishable from MS-H by sequence analysis of variable lipoprotein haemagglutinin (vlhA) gene. Single nucleotide polymorphisms (SNPs) in the obg of MS-H have been found to associate with ts phenotype. In this study, four PCRs followed by high-resolution melting (HRM)-curve analysis of the regions encompassing these SNPs were developed and evaluated for their potential to differentiate MS-H from 36 M. synoviae strains/isolates. The nested-obg PCR-HRM differentiated ts + MS-H vaccine not only from field M. synoviae strains/isolates but also from ts – MS-H revertants. The mean genotype confidence percentages, 96.9±3.4 and 8.8±11.2 for ts + and ts – strains, respectively, demonstrated high differentiating power of the nested-obg PCR-HRM. Using a combination of nested-obg and obg-F3R3 PCR-HRM, 97% of the isolates/strains were typed according to their ts phenotype with all MS-H isolates typed as MS-H. A set of respiratory swabs from MS-H vaccinated specific pathogen free chickens and M. synoviae infected commercial chicken flocks were tested using obg PCR-HRM system and results were consistent with those of vlhA genotyping. The PCR-HRM system developed in this study, proved to be a rapid and reliable tool using pure M. synoviae cultures as well as direct clinical specimens. PMID:24643035

  2. A nested real-time PCR assay for the quantification of Plasmodium falciparum DNA extracted from dried blood spots.

    PubMed

    Tran, Tuan M; Aghili, Amirali; Li, Shanping; Ongoiba, Aissata; Kayentao, Kassoum; Doumbo, Safiatou; Traore, Boubacar; Crompton, Peter D

    2014-10-04

    As public health efforts seek to eradicate malaria, there has been an emphasis on eliminating low-density parasite reservoirs in asymptomatic carriers. As such, diagnosing submicroscopic Plasmodium infections using PCR-based techniques has become important not only in clinical trials of malaria vaccines and therapeutics, but also in active malaria surveillance campaigns. However, PCR-based quantitative assays that rely on nucleic acid extracted from dried blood spots (DBS) have demonstrated lower sensitivity than assays that use cryopreserved whole blood as source material. The density of Plasmodium falciparum asexual parasites was quantified using genomic DNA extracted from dried blood spots (DBS) and the sensitivity of two approaches was compared: quantitative real-time PCR (qPCR) targeting the P. falciparum 18S ribosomal RNA gene, either with an initial conventional PCR amplification prior to qPCR (nested qPCR), or without an initial amplification (qPCR only). Parasite densities determined by nested qPCR, qPCR only, and light microscopy were compared. Nested qPCR results in 10-fold higher sensitivity (0.5 parasites/μl) when compared to qPCR only (five parasites/ul). Among microscopy-positive samples, parasite densities calculated by nested qPCR correlated strongly with microscopy for both asymptomatic (Pearson's r=0.58, P<0.001) and symptomatic (Pearson's r=0.70, P<0.0001) P. falciparum infections. Nested qPCR improves the sensitivity for the detection of P. falciparum blood-stage infection from clinical DBS samples. This approach may be useful for active malaria surveillance in areas where submicroscopic asymptomatic infections are prevalent.

  3. A multiplex nested PCR for the detection and identification of Candida species in blood samples of critically ill paediatric patients

    PubMed Central

    2014-01-01

    Background Nosocomial candidaemia is associated with high mortality rates in critically ill paediatric patients; thus, the early detection and identification of the infectious agent is crucial for successful medical intervention. The PCR-based techniques have significantly increased the detection of Candida species in bloodstream infections. In this study, a multiplex nested PCR approach was developed for candidaemia detection in neonatal and paediatric intensive care patients. Methods DNA samples from the blood of 54 neonates and children hospitalised in intensive care units with suspected candidaemia were evaluated by multiplex nested PCR with specific primers designed to identify seven Candida species, and the results were compared with those obtained from blood cultures. Results The multiplex nested PCR had a detection limit of four Candida genomes/mL of blood for all Candida species. Blood cultures were positive in 14.8% of patients, whereas the multiplex nested PCR was positive in 24.0% of patients, including all culture-positive patients. The results obtained with the molecular technique were available within 24 hours, and the assay was able to identify Candida species with 100% of concordance with blood cultures. Additionally, the multiplex nested PCR detected dual candidaemia in three patients. Conclusions Our proposed PCR method may represent an effective tool for the detection and identification of Candida species in the context of candidaemia diagnosis in children, showing highly sensitive detection and the ability to identify the major species involved in this infection. PMID:25047415

  4. A multiplex nested PCR for the detection and identification of Candida species in blood samples of critically ill paediatric patients.

    PubMed

    Taira, Cleison Ledesma; Okay, Thelma Suely; Delgado, Artur Figueiredo; Ceccon, Maria Esther Jurfest Rivero; de Almeida, Margarete Teresa Gottardo; Del Negro, Gilda Maria Barbaro

    2014-07-21

    Nosocomial candidaemia is associated with high mortality rates in critically ill paediatric patients; thus, the early detection and identification of the infectious agent is crucial for successful medical intervention. The PCR-based techniques have significantly increased the detection of Candida species in bloodstream infections. In this study, a multiplex nested PCR approach was developed for candidaemia detection in neonatal and paediatric intensive care patients. DNA samples from the blood of 54 neonates and children hospitalised in intensive care units with suspected candidaemia were evaluated by multiplex nested PCR with specific primers designed to identify seven Candida species, and the results were compared with those obtained from blood cultures. The multiplex nested PCR had a detection limit of four Candida genomes/mL of blood for all Candida species. Blood cultures were positive in 14.8% of patients, whereas the multiplex nested PCR was positive in 24.0% of patients, including all culture-positive patients. The results obtained with the molecular technique were available within 24 hours, and the assay was able to identify Candida species with 100% of concordance with blood cultures. Additionally, the multiplex nested PCR detected dual candidaemia in three patients. Our proposed PCR method may represent an effective tool for the detection and identification of Candida species in the context of candidaemia diagnosis in children, showing highly sensitive detection and the ability to identify the major species involved in this infection.

  5. Evaluation of a nested-PCR for mycobacterium tuberculosis detection in blood and urine samples.

    PubMed

    da Cruz, Heidi Lacerda Alves; de Albuquerque Montenegro, Rosana; de Araújo Lima, Juliana Falcão; da Rocha Poroca, Diogo; da Costa Lima, Juliana Figueirêdo; Maria Lapa Montenegro, Lílian; Crovella, Sergio; Charifker Schindler, Haiana

    2011-01-01

    The polymerase chain reaction (PCR) and its variations, such as the nested-PCR, have been described as promising techniques for rapid diagnosis of tuberculosis (TB). With the aim of evaluating the usefulness of a nested-PCR method on samples of blood and urine of patients suspected of tuberculosis we analyzed 192 clinical samples, using as a molecular target the insertion element IS6110 specific of M. tuberculosis genome. Nested-PCR method showed higher sensitivity in patients with extrapulmonary tuberculosis (47.8% and 52% in blood and urine) when compared to patients with the pulmonary form of the disease (sensitivity of 29% and 26.9% in blood and urine), regardless of the type of biological sample used. The nested-PCR is a rapid technique that, even if not showing a good sensitivity, should be considered as a helpful tool especially in the extrapulmonary cases or in cases where confirmatory diagnosis is quite difficult to be achieved by routine methods. The performance of PCR-based techniques should be considered and tested in future works on other types of biological specimens besides sputum, like blood and urine, readily obtainable in most cases. The improving of M. tuberculosis nested-PCR detection in TB affected patients will give the possibility of an earlier detection of bacilli thus interrupting the transmission chain of the disease.

  6. Plasticity of parental care under the risk of predation: how much should parents reduce care?

    USGS Publications Warehouse

    Martin, Thomas E.

    2013-01-01

    Predation can be an important agent of natural selection shaping parental care behaviours, and can also favour behavioural plasticity. Parent birds often decrease the rate that they visit the nest to provision offspring when perceived risk is high. Yet, the plasticity of such responses may differ among species as a function of either their relative risk of predation, or the mean rate of provisioning. Here, we report parental provisioning responses to experimental increases in the perceived risk of predation. We tested responses of 10 species of bird in north temperate Arizona and subtropical Argentina that differed in their ambient risk of predation. All species decreased provisioning rates in response to the nest predator but not to a control. However, provisioning rates decreased more in species that had greater ambient risk of predation on natural nests. These results support theoretical predictions that the extent of plasticity of a trait that is sensitive to nest predation risk should vary among species in accordance with predation risk.

  7. New PCR diagnostic systems for the detection and quantification of porcine cytomegalovirus (PCMV).

    PubMed

    Morozov, Vladimir A; Morozov, Alexey V; Denner, Joachim

    2016-05-01

    Pigs are frequently infected with porcine cytomegalovirus (PCMV). Infected adult animals may not present with symptoms of disease, and the virus remains latent. However, the virus may be transmitted to human recipients receiving pig transplants. Recently, it was shown that pig-to-non-human-primate xenotransplantations showed 2 to 3 times lower transplant survival when the donor pig was infected with PCMV. Therefore, highly sensitive methods are required to select virus-free pigs and to examine xenotransplants. Seven previously established PCR detection systems targeting the DNA polymerase gene of PCMV were examined by comparison of thermodynamic parameters of oligonucleotides, and new diagnostic nested PCR and real-time PCR systems with improved parameters and high sensitivity were established. The detection limit of conventional PCR was estimated to be 15 copies, and that of the nested PCR was 5 copies. The sensitivity of the real-time PCR with a TaqMan probe was two copies. An equal efficiency of the newly established detection systems was shown by parallel testing of DNA from sera and blood of six pigs, identifying the same animals as PCMV infected. These new diagnostic PCR systems will improve the detection of PCMV and therefore increase the safety of porcine xenotransplants.

  8. Comparison of loop-mediated isothermal amplification (LAMP) and nested-PCR assay targeting the RE and B1 gene for detection of Toxoplasma gondii in blood samples of children with leukaemia.

    PubMed

    Fallahi, Shirzad; Seyyed Tabaei, Seyyed Javad; Pournia, Yadollah; Zebardast, Nozhat; Kazemi, Bahram

    2014-07-01

    Toxoplasmosis diagnosis constitutes an important measure for disease prevention and control. In this paper, a newly described DNA amplification technique, loop-mediated isothermal amplification (LAMP), and nested-PCR targeting the repeated element (RE) and B1 gene, were compared to each other for the detection of Toxoplasma gondii DNA in blood samples of children with leukaemia. One hundred ten blood samples from these patients were analyzed by LAMP and nested-PCR. Out of 50 seropositive samples (IgM+, IgG+), positive results were obtained with 92% and 86% on RE, B1-LAMP and 82% and 68% on RE, B1-nested PCR analyses, respectively. Of the 50 seronegative samples, three, two and one samples were detected positive by RE-LAMP, B1-LAMP and RE-nested PCR assays, respectively, while none were detected positive by B1-nested PCR. None of the 10 IgM-, IgG+ samples was detected positive after testing LAMP and nested-PCR assays in duplicate. This is the first report of a study in which the LAMP method was applied with high sensitivity and efficacy for the diagnosis of T. gonii in blood samples of children with leukaemia. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Analysis of RET promoter CpG island methylation using methylation-specific PCR (MSP), pyrosequencing, and methylation-sensitive high-resolution melting (MS-HRM): impact on stage II colon cancer patient outcome.

    PubMed

    Draht, Muriel X G; Smits, Kim M; Jooste, Valérie; Tournier, Benjamin; Vervoort, Martijn; Ramaekers, Chantal; Chapusot, Caroline; Weijenberg, Matty P; van Engeland, Manon; Melotte, Veerle

    2016-01-01

    Already since the 1990s, promoter CpG island methylation markers have been considered promising diagnostic, prognostic, and predictive cancer biomarkers. However, so far, only a limited number of DNA methylation markers have been introduced into clinical practice. One reason why the vast majority of methylation markers do not translate into clinical applications is lack of independent validation of methylation markers, often caused by differences in methylation analysis techniques. We recently described RET promoter CpG island methylation as a potential prognostic marker in stage II colorectal cancer (CRC) patients of two independent series. In the current study, we analyzed the RET promoter CpG island methylation of 241 stage II colon cancer patients by direct methylation-specific PCR (MSP), nested-MSP, pyrosequencing, and methylation-sensitive high-resolution melting (MS-HRM). All primers were designed as close as possible to the same genomic region. In order to investigate the effect of different DNA methylation assays on patient outcome, we assessed the clinical sensitivity and specificity as well as the association of RET methylation with overall survival for three and five years of follow-up. Using direct-MSP and nested-MSP, 12.0 % (25/209) and 29.6 % (71/240) of the patients showed RET promoter CpG island methylation. Methylation frequencies detected by pyrosequencing were related to the threshold for positivity that defined RET methylation. Methylation frequencies obtained by pyrosequencing (threshold for positivity at 20 %) and MS-HRM were 13.3 % (32/240) and 13.8 % (33/239), respectively. The pyrosequencing threshold for positivity of 20 % showed the best correlation with MS-HRM and direct-MSP results. Nested-MSP detected RET promoter CpG island methylation in deceased patients with a higher sensitivity (33.1 %) compared to direct-MSP (10.7 %), pyrosequencing (14.4 %), and MS-HRM (15.4 %). While RET methylation frequencies detected by nested-MSP, pyrosequencing, and MS-HRM varied, the prognostic effect seemed similar (HR 1.74, 95 % CI 0.97-3.15; HR 1.85, 95 % CI 0.93-3.86; HR 1.83, 95 % CI 0.92-3.65, respectively). Our results show that upon optimizing and aligning four RET methylation assays with regard to primer location and sensitivity, differences in methylation frequencies and clinical sensitivities are observed; however, the effect on the marker's prognostic outcome is minimal.

  10. Diet of yellow-billed loons (Gavia adamsii) in Arctic lakes during the nesting season inferred from fatty acid analysis

    USGS Publications Warehouse

    Haynes, T B; Schmutz, Joel A.; Bromaghin, Jeffrey F.; Iverson, S J; Padula, V. M.; Rosenberger, A E

    2015-01-01

    Understanding the dietary habits of yellow-billed loons (Gavia adamsii) can give important insights into their ecology, however, studying the diet of loons is difficult when direct observation or specimen collection is impractical. We investigate the diet of yellow-billed loons nesting on the Arctic Coastal Plain of Alaska using quantitative fatty acid signature analysis. Tissue analysis from 26 yellow-billed loons and eleven prey groups (nine fish species and two invertebrate groups) from Arctic lakes suggests that yellow-billed loons are eating high proportions of Alaska blackfish (Dallia pectoralis), broad whitefish (Coregonus nasus) and three-spined stickleback (Gasterosteus aculeatus) during late spring and early summer. The prominence of blackfish in diets highlights the widespread availability of blackfish during the early stages of loon nesting, soon after spring thaw. The high proportions of broad whitefish and three-spined stickleback may reflect a residual signal from the coastal staging period prior to establishing nesting territories on lakes, when loons are more likely to encounter these species. Our analyses were sensitive to the choice of calibration coefficient based on data from three different species, indicating the need for development of loon-specific coefficients for future study and confirmation of our results. Regardless, fish that are coastally distributed and that successfully overwinter in lakes are likely key food items for yellow-billed loons early in the nesting season.

  11. Vulnerability of shortgrass prairie bird assemblages to climate change

    USGS Publications Warehouse

    Skagen, Susan K.; Dreitz, Victoria; Conrey, Reesa Y.; Yackel, Amy; Panjabi, Arvind O.; Knuffman, Lekha

    2016-01-01

    The habitats and resources needed to support grassland birds endemic to North American prairie ecosystems are seriously threatened by impending climate change. To assess the vulnerability of grassland birds to climate change, we consider various components of vulnerability, including sensitivity, exposure, and adaptive capacity (Glick et al. 2011). Sensitivity encompasses the innate characteristics of a species and, in this context, is related to a species’ tolerance to changes in weather patterns. Groundnesting birds, including prairie birds, are particularly responsive to heat waves combined with drought conditions, as revealed by abundance and distribution patterns (Albright et al. 2010). To further assess sensitivity, we estimated reproductive parameters of nearly 3000 breeding attempts of a suite of prairie birds relative to prevailing weather. Fluctuations in weather conditions in eastern Colorado, 1997-2014, influenced breeding performance of a suite of avian species endemic to the shortgrass prairie, many of which have experienced recent population declines. High summer temperatures and intense rain events corresponded with lower nest survival for most species. Although dry conditions favored nest survival of Burrowing Owls and Mountain Plovers (Conrey 2010, Dreitz et al. 2012), drought resulted in smaller clutch sizes and lower nest survival for passerines (Skagen and Yackel Adams 2012, Conrey et al. in review). Declining summer precipitation may reduce the likelihood that some passerine species can maintain stable breeding populations in this region of the shortgrass prairie.

  12. Morphological Manifestations of Parvovirus B19 Infection in the Bone Marrow.

    DTIC Science & Technology

    1997-01-09

    Parvovirus B19 (PV B19 ) preferentially infects erythroid progenitor cells in the bone marrow, frequently causing anemia along with transient aplastic...infection. We devised a highly sensitive two-round, nested PCR procedure to detect PV B19 . Eight of 78 clinical specimens from individuals with

  13. Evaluation of a new single-tube multiprobe real-time PCR for diagnosis of Entamoeba histolytica and Entamoeba dispar.

    PubMed

    Liang, Shih-Yu; Hsia, Kan-Tai; Chan, Yun-Hsien; Fan, Chia-Kwung; Jiang, Donald Dah-Shyong; Landt, Olfert; Ji, Dar-Der

    2010-08-01

    A single-tube multiprobe real-time PCR assay for simultaneous detection of Entamoeba histolytica and Entamoeba dispar was developed. One primer pair with 2 species-specific probes was designed based on new SSU RNA regions of the ribosomal DNA-containing episome. The sensitivity is 1 parasite per milliliter of feces and thus superior to the conventional nested PCR and comparable to other published real-time PCR protocols. The applicability for clinical diagnosis was validated with 218 stool specimens from patients. A total of 51 E. histolytica and 39 E. dispar positive samples was detected by the multiprobe real-time PCR compared to 39 and 22 by routine nested PCR diagnosis. The detection rate of Entamoeba species for the multiprobe real-time PCR assays was significantly higher than the nested PCR (40.8% vs. 28.0%, P < 0.01). The test did not show cross reactivity with DNA from Entamoeba moshkovskii, Giardia lamblia , Cryptosporidium sp., Escherichia coli , or other nonpathogenic enteric parasites. The multiprobe real-time PCR assay is simple and rapid and has high specificity and sensitivity. The assay could streamline the laboratory diagnosis procedure and facilitate epidemiological investigation.

  14. Statistical and procedural issues in the use of heated taxidermic mounts

    USGS Publications Warehouse

    Bakken, G.S.; Kenow, K.P.; Korschgen, C.E.; Boysen, A.F.

    2000-01-01

    Studies using mounts have an inherently nested error structure; calibration and standardization should use the appropriate procedures and statistics. One example is that individual mount differences are nested within morphological factors related to species, age, or gender; without replication, mount differences may be confused with differences due to morphology. Also, the sensitivity of mounts to orientation to wind or sun is nested within mount; without replication, inadvertent variation in mount positioning may be confused with differences among mounts. Data on heat loss from a of 1-day-old mallard duckling mount are used to illustrate orientation sensitivity. (C) 2000 Elsevier Science Ltd. All rights reserved.

  15. Molecular analysis of dolphin morbillivirus: A new sensitive detection method based on nested RT-PCR.

    PubMed

    Centelleghe, Cinzia; Beffagna, Giorgia; Zanetti, Rossella; Zappulli, Valentina; Di Guardo, Giovanni; Mazzariol, Sandro

    2016-09-01

    Cetacean Morbillivirus (CeMV) has been identified as the most pathogenic virus for cetaceans. Over the past three decades, this RNA virus has caused several outbreaks of lethal disease in odontocetes and mysticetes worldwide. Isolation and identification of CeMV RNA is very challenging in whales because of the poor preservation status frequently shown by tissues from stranded animals. Nested reverse transcription polymerase chain reaction (nested RT-PCR) is used instead of conventional RT-PCR when it is necessary to increase the sensitivity and the specificity of the reaction. This study describes a new nested RT-PCR technique useful to amplify small amounts of the cDNA copy of Cetacean morbillivirus (CeMV) when it is present in scant quantity in whales' biological specimens. This technique was used to analyze different tissues (lung, brain, spleen and other lymphoid tissues) from one under human care seal and seven cetaceans stranded along the Italian coastline between October 2011 and September 2015. A well-characterized, 200 base pair (bp) fragment of the dolphin Morbillivirus (DMV) haemagglutinin (H) gene, obtained by nested RT-PCR, was sequenced and used to confirm DMV positivity in all the eight marine mammals under study. In conclusion, this nested RT-PCR protocol can represent a sensitive detection method to identify CeMV-positive, poorly preserved tissue samples. Furthermore, this is also a rather inexpensive molecular technique, relatively easy to apply. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. The efficacy of a nested PCR in detecting cytochrome c oxidase subunit 1 gene of Sarcoptes scabiei var. Hominis for diagnosing scabies.

    PubMed

    Hahm, J E; Kim, C W; Kim, S S

    2018-04-06

    A widespread scabies infestation, associated to long-term residence in nursing homes, is becoming a serious issue in developed countries. Mineral oil examination is regarded as the gold standard in diagnosing scabies, but the sensitivity of this method is generally low-approximately 50%. Molecular tests may contribute to enhance the sensitivity of current tests for laboratory diagnosis of human scabies. In this study, we developed new primers for a nested PCR for the cytochrome c oxidase subunit 1 (cox1) gene of Sarcoptes scabiei var. hominis to increase the sensitivity of a previously developed conventional PCR. Clinically suspected scabies patients underwent dermoscopy-guided skin scraping with microscopic examination. The diagnosis was positive for scabies when mites or eggs were found under the microscope, and patients were then designated as 'microscopy-positive'. Patients in the 'microscopy-negative' group presented with negative microscopic results. Skin scrapings were collected from both groups for PCR. Of the total 63 samples, 28 were microscopy-positive and 35 were negative with no differences in sex and age between the two groups. All microscopically proven scabies cases were positive with the cox1 nested PCR. Among microscopy-negative ones, S. scabiei DNA was detected in 9 samples. If sensitivity of the cox1 nested PCR is considered 100% (95% CI, 90.51-100), then sensitivity of microscopy is 75.68% (95% CI, 58.80-88.23; P = 0.004). Nested PCR can be successfully used as an alternative method for diagnosing suspected scabies patient. Therefore, infection control measures and treatments can be initiated before significant transmission occurs, minimizing the risk of outbreaks. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  17. Development of polymerase chain reaction-based diagnostic tests for detection of Malsoor virus & adenovirus isolated from Rousettus species of bats in Maharashtra, India.

    PubMed

    Shete, Anita M; Yadav, Pragya; Kumar, Vimal; Nikam, Tushar; Mehershahi, Kurosh; Kokate, Prasad; Patil, Deepak; Mourya, Devendra T

    2017-01-01

    Bats are recognized as important reservoirs for emerging infectious disease and some unknown viral diseases. Two novel viruses, Malsoor virus (family Bunyaviridae, genus, Phlebovirus) and a novel adenovirus (AdV) (family, Adenoviridae genus, Mastadenovirus), were identified from Rousettus bats in the Maharashtra State of India. This study was done to develop and optimize real time reverse transcription - polymerase chain reaction (RT-PCR) assays for Malsoor virus and real time and nested PCR for adenovirus from Rousettus bats. For rapid and accurate screening of Malsoor virus and adenovirus a nested polymerase chain reaction and TaqMan-based real-time PCR were developed. Highly conserved region of nucleoprotein gene of phleboviruses and polymerase gene sequence from the Indian bat AdV isolate polyprotein gene were selected respectively for diagnostic assay development of Malsoor virus and AdV. Sensitivity and specificity of assays were calculated and optimized assays were used to screen bat samples. Molecular diagnostic assays were developed for screening of Malsoor virus and AdV and those were found to be specific. Based on the experiments performed with different parameters, nested PCR was found to be more sensitive than real-time PCR; however, for rapid screening, real-time PCR can be used and further nested PCR can be used for final confirmation or in those laboratories where real-time facility/expertise is not existing. This study reports the development and optimization of nested RT-PCR and a TaqMan-based real-time PCR for Malsoor virus and AdV. The diagnostic assays can be used for rapid detection of these novel viruses to understand their prevalence among bat population.

  18. Detection of Mycobacterium bovis in Bovine and Bubaline Tissues Using Nested-PCR for TbD1

    PubMed Central

    Araújo, Cristina P.; Osório, Ana Luiza A. R.; Jorge, Kláudia S. G.; Ramos, Carlos Alberto N.; Filho, Antonio Francisco S.; Vidal, Carlos Eugênio S.; Roxo, Eliana; Nishibe, Christiane; Almeida, Nalvo F.; Júnior, Antônio A. F.; Silva, Marcio R.; Neto, José Diomedes B.; Cerqueira, Valíria D.; Zumárraga, Martín J.; Araújo, Flábio R.

    2014-01-01

    In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis. PMID:24618787

  19. Detection of Mycobacterium bovis in bovine and bubaline tissues using nested-PCR for TbD1.

    PubMed

    Araújo, Cristina P; Osório, Ana Luiza A R; Jorge, Kláudia S G; Ramos, Carlos Alberto N; Filho, Antonio Francisco S; Vidal, Carlos Eugênio S; Roxo, Eliana; Nishibe, Christiane; Almeida, Nalvo F; Júnior, Antônio A F; Silva, Marcio R; Neto, José Diomedes B; Cerqueira, Valíria D; Zumárraga, Martín J; Araújo, Flábio R

    2014-01-01

    In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

  20. Detection of canine distemper virus (CDV) through one step RT-PCR combined with nested PCR.

    PubMed

    Kim, Y H; Cho, K W; Youn, H Y; Yoo, H S; Han, H R

    2001-04-01

    A one step reverse transcription PCR (RT-PCR) combined nested PCR was set up to increase efficiency in the diagnosis of canine distemper virus (CDV) infection after developement of nested PCR. Two PCR primer sets were designed based on the sequence of nucleocapsid gene of CDV Onderstepoort strain. One-step RT-PCR with the outer primer pair was revealed to detect 10(2) PFU/ml. The sensitivity was increased hundredfold using the one-step RT-PCR combined with the nested PCR. Specificity of the PCR was also confirmed using other related canine virus and peripheral blood mononuclear cells (PBMC) and body secretes of healthy dogs. Of the 51 blood samples from dogs clinically suspected of CD, 45 samples were revealed as positive by one-step RT-PCR combined with nested PCR. However, only 15 samples were identified as positive with a single one step RT-PCR. Therefore approximately 60% increase in the efficiency of the diagnosis was observed by the combined method. These results suggested that one step RT-PCR combined with nested PCR could be a sensitive, specific, and practical method for diagnosis of CDV infection.

  1. Floristic conservation value, nested understory floras, and the development of second-growth forest.

    PubMed

    Spyreas, Greg; Matthews, Jeffrey W

    2006-08-01

    Nestedness analysis can reveal patterns of plant composition and diversity among forest patches. For nested floral assemblages, the plants occupying any one patch are a nested subset of the plants present in successively more speciose patches. Elimination of sensitive understory plants with human disturbance is one of several mechanisms hypothesized to generate nonrandom, nested floral distributions. Hypotheses explaining distributions of understory plants remain unsubstantiated across broad landscapes of varying forest types and disturbance histories. We sampled the vegetation of 51 floodplain and 55 upland forests across Illinois (USA) to examine how the diversity, composition, and nestedness of understory floras related to their overstory growth and structure (basal area), and their overall floristic conservation value (mean C). We found that plant assemblages were nested with respect to site species richness, such that rare plants indicated diverse forests. Floras were also nested with respect to site mean C and basal area (BA). However, in an opposite pattern from what we had expected, floras of high-BA stands were nested subsets of those of low-BA stands. A set of early-successional plants restricted to low-BA stands, and more importantly, the absence of a set of true forest plants in high-BA stands, accounted for this pattern. Additionally, we observed a decrease in species richness with increasing BA. These results are consistent with the hypothesis that recovery of true forest plants does not occur concurrently with overstory regeneration following massive anthropogenic disturbance. Nestedness by site mean C indicates that high conservation value (conservative) plants co-occur in highly diverse stands; these forests are assumed to be less disturbed historically. Because site mean C was uncorrelated with BA, BA-neutral disturbances such as livestock usage are suggested as accounting for between-site differences in mean C. When considered individually, conservative plants were actually more likely to be found in low-BA stands (uplands only). This suggests that floras of historically more open-canopied oak-hickory uplands are being degraded by canopy closure from increasing density of "mesophytic, nonpyrogenic" trees. It also indirectly suggests that recent moderate logging is uncorrelated with floristic conservation values.

  2. A rapid single-tube protocol for HAV detection by nested real-time PCR.

    PubMed

    Hu, Yuan; Arsov, Ivica

    2014-09-01

    Infections by food-borne viruses such as hepatitis A virus (HAV) and norovirus are significant public health concerns worldwide. Since food-borne viruses are rarely confirmed through direct isolation from contaminated samples, highly sensitive molecular techniques remain the methods of choice for the detection of viral genetic material. Our group has previously developed a specific nested real-time PCR (NRT-PCR) assay for HAV detection that improved overall sensitivity. Furthermore in this study, we have developed a single-tube NRT-PCR approach for HAV detection in food samples that reduces the likelihood of cross contamination between tubes during sample manipulation. HAV RNA was isolated from HAV-spiked food samples and HAV-infected cell cultures. All reactions following HAV RNA isolation, including conventional reverse transcriptase PCR, nested-PCR, and RT-PCR were performed in a single tube. Our results demonstrated that all the samples tested positive by RT-PCR and nested-PCR were also positive by a single-tube NRT-PCR. The detection limits observed for HAV-infected cell cultures and HAV-spiked green onions were 0.1 and 1 PFU, respectively. This novel method retained the specificity and robustness of the original NRT-PCR method, while greatly reducing sample manipulation, turnaround time, and the risk of carry-over contamination. Single-tube NRT-PCR thus represents a promising new tool that can potentially facilitate the detection of HAV in foods thereby improving food safety and public health.

  3. Evaluation of efficiency of nested multiplex allele-specific PCR assay for detection of multidrug resistant tuberculosis directly from sputum samples.

    PubMed

    Mistri, S K; Sultana, M; Kamal, S M M; Alam, M M; Irin, F; Nessa, J; Ahsan, C R; Yasmin, M

    2016-05-01

    For an effective control of tuberculosis, rapid detection of multidrug resistant tuberculosis (MDR-TB) is necessary. Therefore, we developed a modified nested multiplex allele-specific polymerase chain reaction (MAS-PCR) method that enables rapid MDR-TB detection directly from sputum samples. The efficacy of this method was evaluated using 79 sputum samples collected from suspected tuberculosis patients. The performance of nested MAS-PCR method was compared with other MDR-TB detection methods like drug susceptibility testing (DST) and DNA sequencing. As rifampicin (RIF) resistance conforms to MDR-TB in greater than 90% cases, only the presence of RIF-associated mutations in rpoB gene was determined by DNA sequencing and nested MAS-PCR to detect MDR-TB. The concordance between nested MAS-PCR and DNA sequencing results was found to be 96·3%. When compared with DST, the sensitivity and specificity of nested MAS-PCR for RIF-resistance detection were determined to be 92·9 and 100% respectively. For developing- and high-TB burden countries, molecular-based tests have been recommended by the World Health Organization for rapid detection of MDR-TB. The results of this study indicate that, nested MAS-PCR assay might be a practical and relatively cost effective molecular method for rapid detection of MDR-TB from suspected sputum samples in developing countries with resource poor settings. © 2016 The Society for Applied Microbiology.

  4. Novel insights into the ontogeny of nestmate recognition in Polistes social wasps.

    PubMed

    Signorotti, Lisa; Cappa, Federico; d'Ettorre, Patrizia; Cervo, Rita

    2014-01-01

    The importance of early experience in animals' life is unquestionable, and imprinting-like phenomena may shape important aspects of behaviour. Early learning typically occurs during a sensitive period, which restricts crucial processes of information storage to a specific developmental phase. The characteristics of the sensitive period have been largely investigated in vertebrates, because of their complexity and plasticity, both in behaviour and neurophysiology, but early learning occurs also in invertebrates. In social insects, early learning appears to influence important social behaviours such as nestmate recognition. Yet, the mechanisms underlying recognition systems are not fully understood. It is currently believed that Polistes social wasps are able to discriminate nestmates from non-nestmates following the perception of olfactory cues present on the paper of their nest, which are learned during a strict sensitive period, immediately after emergence. Here, through differential odour experience experiments, we show that workers of Polistes dominula develop correct nestmate recognition abilities soon after emergence even in absence of what have been so far considered the necessary cues (the chemicals spread on nest paper). P. dominula workers were exposed for the first four days of adult life to paper fragments from their nest, or from a foreign conspecific nest or to a neutral condition. Wasps were then transferred to their original nests where recognition abilities were tested. Our results show that wasps do not alter their recognition ability if exposed only to nest material, or in absence of nest material, during the early phase of adult life. It thus appears that the nest paper is not used as a source of recognition cues to be learned in a specific time window, although we discuss possible alternative explanations. Our study provides a novel perspective for the study of the ontogeny of nestmate recognition in Polistes wasps and in other social insects.

  5. Differential behavioural and endocrine responses of common voles (Microtus arvalis) to nest predators and resource competitors.

    PubMed

    Liesenjohann, Monique; Liesenjohann, Thilo; Palme, Rupert; Eccard, Jana Anja

    2013-09-08

    Adaptive behavioural strategies promoting co-occurrence of competing species are known to result from a sympatric evolutionary past. Strategies should be different for indirect resource competition (exploitation, e.g., foraging and avoidance behaviour) than for direct interspecific interference (e.g., aggression, vigilance, and nest guarding). We studied the effects of resource competition and nest predation in sympatric small mammal species using semi-fossorial voles and shrews, which prey on vole offspring during their sensitive nestling phase. Experiments were conducted in caged outdoor enclosures. Focus common vole mothers (Microtus arvalis) were either caged with a greater white-toothed shrew (Crocidura russula) as a potential nest predator, with an herbivorous field vole (Microtus agrestis) as a heterospecific resource competitor, or with a conspecific resource competitor. We studied behavioural adaptations of vole mothers during pregnancy, parturition, and early lactation, specifically modifications of the burrow architecture and activity at burrow entrances. Further, we measured pre- and postpartum faecal corticosterone metabolites (FCMs) of mothers to test for elevated stress hormone levels. Only in the presence of the nest predator were prepartum FCMs elevated, but we found no loss of vole nestlings and no differences in nestling body weight in the presence of the nest predator or the heterospecific resource competitor. Although the presence of both the shrew and the field vole induced prepartum modifications to the burrow architecture, only nest predators caused an increase in vigilance time at burrow entrances during the sensitive nestling phase. Voles displayed an adequate behavioural response for both resource competitors and nest predators. They modified burrow architecture to improve nest guarding and increased their vigilance at burrow entrances to enhance offspring survival chances. Our study revealed differential behavioural adaptations to resource competitors and nest predators.

  6. Differential behavioural and endocrine responses of common voles (Microtus arvalis) to nest predators and resource competitors

    PubMed Central

    2013-01-01

    Background Adaptive behavioural strategies promoting co-occurrence of competing species are known to result from a sympatric evolutionary past. Strategies should be different for indirect resource competition (exploitation, e.g., foraging and avoidance behaviour) than for direct interspecific interference (e.g., aggression, vigilance, and nest guarding). We studied the effects of resource competition and nest predation in sympatric small mammal species using semi-fossorial voles and shrews, which prey on vole offspring during their sensitive nestling phase. Experiments were conducted in caged outdoor enclosures. Focus common vole mothers (Microtus arvalis) were either caged with a greater white-toothed shrew (Crocidura russula) as a potential nest predator, with an herbivorous field vole (Microtus agrestis) as a heterospecific resource competitor, or with a conspecific resource competitor. Results We studied behavioural adaptations of vole mothers during pregnancy, parturition, and early lactation, specifically modifications of the burrow architecture and activity at burrow entrances. Further, we measured pre- and postpartum faecal corticosterone metabolites (FCMs) of mothers to test for elevated stress hormone levels. Only in the presence of the nest predator were prepartum FCMs elevated, but we found no loss of vole nestlings and no differences in nestling body weight in the presence of the nest predator or the heterospecific resource competitor. Although the presence of both the shrew and the field vole induced prepartum modifications to the burrow architecture, only nest predators caused an increase in vigilance time at burrow entrances during the sensitive nestling phase. Conclusion Voles displayed an adequate behavioural response for both resource competitors and nest predators. They modified burrow architecture to improve nest guarding and increased their vigilance at burrow entrances to enhance offspring survival chances. Our study revealed differential behavioural adaptations to resource competitors and nest predators. PMID:24010574

  7. An alternative nested-PCR assay for the detection of Toxoplasma gondii strains based on GRA7 gene sequences.

    PubMed

    Costa, Maria Eduarda S M; Oliveira, Claudio Bruno S; Andrade, Joelma Maria de A; Medeiros, Thatiany A; Neto, Valter F Andrade; Lanza, Daniel C F

    2016-07-01

    Toxoplasma gondii is a widespread parasite able to infect virtually any nucleated cells of warm-blooded hosts. In some cases, T. gondii detection using already developed PCR primers can be inefficient in routine laboratory tests, especially to detect atypical strains. Here we report a new nested-PCR protocol able to detect virtually all T. gondii isolates. Analyzing 685 sequences available in GenBank, we determine that GRA7 is one of the most conserved genes of T. gondii genome. Based on an alignment of 85 GRA7 sequences new primer sets that anneal in the highly conserved regions of this gene were designed. The new GRA7 nested-PCR assay providing sensitivity and specificity equal to or greater than the gold standard PCR assays for T. gondii detection, that amplify the B1 sequence or the repetitive 529bp element. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Gamma-Ray Focusing Optics for Small Animal Imaging

    NASA Technical Reports Server (NTRS)

    Pivovaroff, M. J.; Barber, W. C.; Craig, W. W.; Hasegawa, B. H.; Ramsey, B. D.; Taylor, C.

    2004-01-01

    There is a well-established need for high-resolution radionuclide imaging techniques that provide non-invasive measurement of physiological function in small animals. We, therefore, have begun developing a small animal radionuclide imaging system using grazing incidence mirrors to focus low-energy gamma-rays emitted by I-125, and other radionuclides. Our initial prototype optic, fabricated from thermally-formed glass, demonstrated a resolution of 1500 microns, consistent with the performance predicted by detailed simulations. More recently, we have begun constructing mirrors using a replication technique that reduces low spatial frequency errors in the mirror surface, greatly improving the resolution. Each technique offers particular advantages: e.g., multilayer coatings are easily deposited on glass, while superior resolution is possible with replicated optics. Scaling the results from our prototype optics, which only have a few nested shells, to system where the lens has a full complement of several tens of nested shells, a sensitivity of approx. 1 cps/micro Ci is possible, with the exact number dependent on system magnification and radionuclide species. (Higher levels of efficiency can be obtained with multi-optic imaging systems.) The gamma-ray lens will achieve a resolution as good as 100 microns, independent of the final sensitivity. The combination of high spatial resolution and modest sensitivity will enable in vivo single photon emission imaging studies in small animals.

  9. Development of a nested polymerase chain reaction for amplification of a sequence of the p57 gene of Renibacterium salmoninarum that provides a highly sensitive method for detection of the bacterium in salmonid kidney

    USGS Publications Warehouse

    Chase, D.M.; Pascho, R.J.

    1998-01-01

    Nucleic acid-based assays have shown promise for diagnosing Renibacterium salmoninarum in tissues and body fluids of salmonids. DeVelopment of a nested polymerase chain reaction (PCR) method to detect a 320 bp DNA segment of the gene encoding the p57 protein of R. salmoninarum is described. Whereas a conventional PCR for a 383 bp segment of the p57 gene reliably detected 1000 R. salmoninarum cells per reaction in kidney tissue, the nested PCR detected as few as 10 R. salmoninarum per reaction in kidney tissue. Two DNA extraction methods for the nested PCR were compared and the correlation between replicate samples was generally higher in samples extracted by the QIAamp system compared with those extracted by the phenol/chloroform method. The specificity of the nested PCR was confirmed by testing DNA extracts of common bacterial fish pathogens and a panel of bacterial species reported to cause false-positive reactions in the enzyme-linked immunosorbent assay (ELISA) and the fluorescent antibody test (FAT) for R. salmoninarum. Kidney samples from 74 naturally infected chinook Salmon were examined by the nested PCR, the ELISA, and the FAT, and the detected prevalences of R. salmoninarum were 61, 47, and 43%, respectively.

  10. Differentiation of five enterohepatic Helicobacter species by nested PCR with high-resolution melting curve analysis.

    PubMed

    Wu, Miaoli; Rao, Dan; Zhu, Yujun; Wang, Jing; Yuan, Wen; Zhang, Yu; Huang, Ren; Guo, Pengju

    2017-04-01

    Enterohepatic Helicobacter species (EHS) are widespread in rodent species around the world. Several studies have demonstrated that infection with EHS can interfere with the outcomes of animal experiments in cancer research and significantly influence the study results. Therefore, it is essential to establish a rapid detection and identification of EHS for biomedical research using laboratory rodents. Our study aimed to develop a rapid and sensitive method to detect and distinguish five enterohepatic Helicobacter species. Nested PCR followed by high-resolution melting curve analysis (HRM) was developed for identification of H. bilis, H. rodentium, H. muridarum, H. typhlonius, as well as H. hepaticus. To validate the accuracy of nested PCR-HRM analysis, quantitative real-time PCR methods for five different enterohepatic Helicobacter species were developed. A total of 50 cecal samples were tested using both nested PCR-HRM analysis and qPCR method. The nested PCR-HRM method could distinguish five enterohepatic Helicobacter species by different melting temperatures. The melting curve were characterized by peaks of 78.7 ± 0.12°C for H. rodentium, 80.51 ± 0.09°C for H. bilis, 81.6 ± 0.1°C for H. typhlonius, 82.11 ± 0.18°C for H. muridarum, and 82.95 ± 0.09°C for H. hepaticus. The nested PCR-HRM assay is a simple, rapid, and cost-effective assay. This assay could be a useful tool for molecular epidemiology study of enterohepatic Helicobacter infection and an attractive alternative for genotyping of enterohepatic Helicobacter species. © 2016 John Wiley & Sons Ltd.

  11. Protecting unknown two-qubit entangled states by nesting Uhrig's dynamical decoupling sequences

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mukhtar, Musawwadah; Soh, Wee Tee; Saw, Thuan Beng

    2010-11-15

    Future quantum technologies rely heavily on good protection of quantum entanglement against environment-induced decoherence. A recent study showed that an extension of Uhrig's dynamical decoupling (UDD) sequence can (in theory) lock an arbitrary but known two-qubit entangled state to the Nth order using a sequence of N control pulses [Mukhtar et al., Phys. Rev. A 81, 012331 (2010)]. By nesting three layers of explicitly constructed UDD sequences, here we first consider the protection of unknown two-qubit states as superposition of two known basis states, without making assumptions of the system-environment coupling. It is found that the obtained decoherence suppression canmore » be highly sensitive to the ordering of the three UDD layers and can be remarkably effective with the correct ordering. The detailed theoretical results are useful for general understanding of the nature of controlled quantum dynamics under nested UDD. As an extension of our three-layer UDD, it is finally pointed out that a completely unknown two-qubit state can be protected by nesting four layers of UDD sequences. This work indicates that when UDD is applicable (e.g., when the environment has a sharp frequency cutoff and when control pulses can be taken as instantaneous pulses), dynamical decoupling using nested UDD sequences is a powerful approach for entanglement protection.« less

  12. Detection of Pneumocystis jirovecii by nested PCR in HIV-negative patients with pulmonary disease.

    PubMed

    Santos, Cristina Rodrigues; de Assis, Ângela M; Luz, Edson A; Lyra, Luzia; Toro, Ivan F; Seabra, José Claudio C; Daldin, Dira H; Marcalto, Tathiane U; Galasso, Marcos T; Macedo, Ronaldo F; Schreiber, Angélica Z; Aoki, Francisco H

    Nested PCR can be used to determine the status of Pneumocystis jirovecii infection in other lung diseases. This study sought to detect a target DNA fragment (mitochondrial large subunit rRNA or mtL SUrRNA) of P. jirovecii in patients with lung disease who underwent bronchoscopy with collection of bronchoalveolar lavage (BAL). The results from toluidine blue staining were compared with those obtained using molecular methods that included an "in house" DNA extraction procedure, PCR and nested PCR. Fifty-five BAL samples from patients with atypical chest X-rays were screened for P. jirovecii. None of the samples was positive for P. jirovecii using toluidine blue staining. In contrast, P. jirovecii DNA was detected by nested PCR in BAL samples from 36 of 55 patients (65.5%). The lung diseases in the patients included cancer, pneumonia, tuberculosis, and chronic obstructive pulmonary disease (COPD). Other chronic problems in the patients included hypertension, diabetes, smoking, and alcoholism. Nested PCR showed high sensitivity for detecting P. jirovecii, especially when compared with toluidine blue staining. Using this method, P. jirovecii infection was detected in HIV-negative patients with lung disease. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  13. Nesting ecology of roseate spoonbills at Nueces Bay, Texas

    USGS Publications Warehouse

    White, Donald H.; Mitchell, Christine A.; Cromartie, E.

    1982-01-01

    We conducted a study in 1978-1980 of the nesting ecology of Roseate Spoonbills (Ajaia ajaja) in a relatively polluted environment at Nueces Bay, Texas. For 154 marked nests, the average clutch size was 3.0 eggs; 73% of the eggs hatched, and 87% of the nests were successful (hatched at least 1 young). The average nest success rate (total fledglings:total eggs) was 50% or 1.5 fledglings per total nests. Incubation began the day after the first egg was laid, and the incubation period for each egg was 22 days. Eggs hatched in the order that they were laid; the first and second eggs hatched on consecutive days, and the second, third, and fourth eggs hatched every other day. Nest composition and size were highly consistent, but nest placement varied considerably and was dependent on the vegetative configuration of the dredge-material islands. Growth rate of nestlings conformed to a 'standard' growth curve, where body weight of nestlings at fledging equaled that of adults. There was no difference in weight gain among siblings based on actual age. Nestlings fledged at about 6 weeks of age, when feather development was complete. At fledging, the bills of juveniles had almost reached adult width, but bill length was only 67% that of adults. In general, environmental pollutants were low (<2 ppm) in spoonbill eggs, though a few eggs contained elevated concentrations of DDE (up to 15 ppm, wet weight). Some eggshells were 5% thinner than those in museum collections, but the degree of thinning was not within the range known to cause population declines. We conclude that organochlorine pollutants are not adversely affecting spoonbill reproduction at Nueces Bay, Texas, though the area is surrounded by industries and agricultural lands. Apparently, spoonbills are less sensitive to these contaminants than are other aquatic species.

  14. Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys.

    PubMed

    Li, Peipei; Zhao, Zhenjun; Wang, Ying; Xing, Hua; Parker, Daniel M; Yang, Zhaoqing; Baum, Elizabeth; Li, Wenli; Sattabongkot, Jetsumon; Sirichaisinthop, Jeeraphat; Li, Shuying; Yan, Guiyun; Cui, Liwang; Fan, Qi

    2014-05-08

    Nested PCR is considered a sensitive and specific method for detecting malaria parasites and is especially useful in epidemiological surveys. However, the preparation of DNA templates for PCR is often time-consuming and costly. A simplified PCR method was developed to directly use a small blood filter paper square (2 × 2 mm) as the DNA template after treatment with saponin. This filter paper-based nested PCR method (FP-PCR) was compared to microscopy and standard nested PCR with DNA extracted by using a Qiagen DNA mini kit from filter paper blood spots of 204 febrile cases. The FP-PCR technique was further applied to evaluate malaria infections in 1,708 participants from cross-sectional epidemiological surveys conducted in Myanmar and Thailand. The FP-PCR method had a detection limit of ~0.2 parasites/μL blood, estimated using cultured Plasmodium falciparum parasites. With 204 field samples, the sensitivity of the FP-PCR method was comparable to that of the standard nested PCR method, which was significantly higher than that of microscopy. Application of the FP-PCR method in large cross-sectional studies conducted in Myanmar and Thailand detected 1.9% (12/638) and 6.2% (66/1,070) asymptomatic Plasmodium infections, respectively, as compared to the detection rates of 1.3% (8/638) and 0.04% (4/1,070) by microscopy. This FP-PCR method was much more sensitive than microscopy in detecting Plasmodium infections. It drastically increased the detection sensitivity of asymptomatic infections in cross-sectional surveys conducted in Thailand and Myanmar, suggesting that this FP-PCR method has a potential for future applications in malaria epidemiology studies.

  15. Nested methylation-specific polymerase chain reaction cancer detection method

    DOEpatents

    Belinsky, Steven A [Albuquerque, NM; Palmisano, William A [Edgewood, NM

    2007-05-08

    A molecular marker-based method for monitoring and detecting cancer in humans. Aberrant methylation of gene promoters is a marker for cancer risk in humans. A two-stage, or "nested" polymerase chain reaction method is disclosed for detecting methylated DNA sequences at sufficiently high levels of sensitivity to permit cancer screening in biological fluid samples, such as sputum, obtained non-invasively. The method is for detecting the aberrant methylation of the p16 gene, O 6-methylguanine-DNA methyltransferase gene, Death-associated protein kinase gene, RAS-associated family 1 gene, or other gene promoters. The method offers a potentially powerful approach to population-based screening for the detection of lung and other cancers.

  16. Nested PCR for specific diagnosis of Taenia solium taeniasis.

    PubMed

    Mayta, Holger; Gilman, Robert H; Prendergast, Emily; Castillo, Janeth P; Tinoco, Yeny O; Garcia, Hector H; Gonzalez, Armando E; Sterling, Charles R

    2008-01-01

    Taeniasis due to Taenia solium is a disease with important public health consequences, since the larval stage is not exclusive to the animal intermediate, the pig, but also infects humans, causing neurocysticercosis. Early diagnosis and treatment of T. solium tapeworm carriers is important to prevent human cysticercosis. Current diagnosis based on microscopic observation of eggs lacks both sensitivity and specificity. In the present study, a nested-PCR assay targeting the Tso31 gene was developed for the specific diagnosis of taeniasis due to T. solium. Initial specificity and sensitivity testing was performed using stored known T. solium-positive and -negative samples. The assay was further analyzed under field conditions by conducting a case-control study of pretreatment stool samples collected from a population in an area of endemicity. Using the archived samples, the assay showed 97% (31/32) sensitivity and 100% (123/123) specificity. Under field conditions, the assay had 100% sensitivity and specificity using microscopy/enzyme-linked immunosorbent assay coproantigen testing as the gold standards. The Tso31 nested PCR described here might be a useful tool for the early diagnosis and prevention of taeniasis/cysticercosis.

  17. Identification of and Screening for Human Helicobacter cinaedi Infections and Carriers via Nested PCR

    PubMed Central

    Oyama, Kohta; Khan, Shahzada; Okamoto, Tatsuya; Fujii, Shigemoto; Ono, Katsuhiko; Matsunaga, Tetsuro; Yoshitake, Jun; Sawa, Tomohiro; Tomida, Junko; Kawamura, Yoshiaki

    2012-01-01

    Helicobacter cinaedi is the most frequently reported enterohepatic Helicobacter species isolated from humans. Earlier research suggested that certain patients with H. cinaedi infection may remain undiagnosed or incorrectly diagnosed because of difficulties in detecting the bacteria by conventional culture methods. Here, we report a nested PCR assay that rapidly detects the cytolethal distending toxin gene (cdt) of H. cinaedi with high specificity and sensitivity. Specificity of the assay was validated by using different species of Helicobacter and Campylobacter, as well as known H. cinaedi-positive and -negative samples. The sensitivity of detection for the cdt gene in the assay was 102 CFU/ml urine or 102 CFU/105 infected RAW 264.7 cells. In an H. cinaedi-infected mouse model, the cdt gene of H. cinaedi was effectively detected via the assay with urine (6/7), stool (2/3), and blood (2/6) samples. Importantly, it detected H. cinaedi in blood, urine, and stool samples from one patient with a suspected H. cinaedi infection and three patients with known infections. The assay was further used clinically to follow up two H. cinaedi-infected patients after antibiotic treatment. Stool samples from these two patients evaluated by nested PCR after antibiotic therapy showed clearance of bacterial DNA. Finally, analysis of stool specimens from healthy volunteers showed occasional positive reactions (4/30) to H. cinaedi DNA, which suggests intestinal colonization by H. cinaedi in healthy subjects. In conclusion, this nested PCR assay may be useful for the rapid diagnosis, antimicrobial treatment evaluation, and epidemiological study of H. cinaedi infection. PMID:23015666

  18. Optimized nested polymerase chain reaction for antemortem detection of Mycobacteria in Amazon parrots (Amazona aestiva) and orange-winged Amazons (Amazona amazonica).

    PubMed

    Baquião, Arianne Costa; Luna, Janaina Oliveira; Medina, Aziz Orro; Sanfilippo, Luiz Francisco; de Faria, Maria Jacinta; dos Santos, Manuel Armando Azevedo

    2014-03-01

    The objectives of this study were to optimize nested polymerase chain reaction (PCR) for Mycobacterium avium complex and Mycobacterium tuberculosis complex and apply them on samples from parrots. Results were negative for the presence of these Mycobacterium in the samples, and nested PCR was specific, faster, and more sensitive than other tests, thereby justifying its use in antemortem diagnosis.

  19. Efficiency of Nested-PCR in Detecting Asymptomatic Cases toward Malaria Elimination Program in an Endemic Area of Iran.

    PubMed

    Turki, Habibollah; Raeisi, Ahmad; Malekzadeh, Kianoosh; Ghanbarnejad, Amin; Zoghi, Samaneh; Yeryan, Masoud; Abedi Nejad, Masoumeh; Mohseni, Fatemeh; Shekari, Mohammad

    2015-01-01

    The aim of this study was to detect low parasite and asymptomatic malaria infections by means of three malaria diagnostic tests, in a low transmission region of Minab district, Hormozgan Province, southern Iran. Blood samples of 200 healthy volunteers from Bagh-e-Malek area were evaluated using microscopic, rapid diagnostic tests (RDT) and nested-PCR to inspect malaria parasite. The results showed no Plasmodium parasite in subjects by means of microscopy and RDT. However, 3 P. vivax positive samples (1.5%) were discovered by Nested-PCR while microscopy and RDT missed the cases. Microscopy as the gold standard method and RDT correctly identified 98.5% of cases, and molecular analysis is sensitive and reliable, especially in the detection of "asymptomatic" infections for active case surveillance. Regarding the existence of asymptomatic malaria in endemic area of Hormozgan, Iran, nested-PCR could be considered as a sensitive tool to interrupt malaria transmission in the country, beside the microscopic and RDT methods.

  20. Identification of individual powdery mildew fungi infecting leaves and direct detection of gene expression by single conidium polymerase chain reaction.

    PubMed

    Matsuda, Yoshinori; Sameshima, Takeshi; Moriura, Nobuyuki; Inoue, Kanako; Nonomura, Teruo; Kakutani, Koji; Nishimura, Hiroaki; Kusakari, Shin-Ichi; Takamatsu, Susumu; Toyoda, Hideyoshi

    2005-10-01

    ABSTRACT Greenhouse-grown tomato seedlings were inoculated naturally with two genera of powdery mildew conidia forming appressorial germ tubes that could not be differentiated by length alone. For direct identification, single germinated conidia were removed from leaves by means of a glass pipette linked to the manipulator of a high-fidelity digital microscope. This microscope enabled in vivo observation of the fungi without leaf decoloration or fungal staining. The isolated conidia were subjected to PCR amplification of the 5.8S rDNA and its adjacent internal transcribed spacer sequences followed by nested PCR to attain sensitivity high enough to amplify target nucleotide sequences (PCR/nested PCR). Target sequences from the conidia were completely coincident with those of the pathogen Oidium neolycopersici or Erysiphe trifolii (syn. Microsphaera trifolii), which is nonpathogenic on tomato. Using RT-PCR/nested PCR or multiplex RT-PCR/nested PCR, it was possible to amplify transcripts expressed in single conidia. Conidia at pre- and postgermination stages were removed individually from tomato leaves, and two powdery mildew genes were monitored. The results indicated that the beta-tubulin homolog TUB2-ol was expressed at pre- and postgermination stages and the cutinase homolog CUT1-ol was only expressed postgermination. Combining digital microscopic micromanipulation and two-step PCR amplification is thus useful for investigation of individual propagules on the surface of plants.

  1. Detection of adenoviruses in shellfish by means of conventional-PCR, nested-PCR, and integrated cell culture PCR (ICC/PCR).

    PubMed

    Rigotto, C; Sincero, T C M; Simões, C M O; Barardi, C R M

    2005-01-01

    We tested three PCR based methodologies to detect adenoviruses associated with cultivated oysters. Conventional-PCR, nested-PCR, and integrated cell culture-PCR (ICC/PCR) were first optimized using oysters seeded with know amounts of Adenovirus serotype 5 (Ad5). The maximum sensitivity for Ad5 detection was determined for each method, and then used to detect natural adenovirus contamination in oysters from three aquiculture farms in Florianopolis, Santa Catarina State, Brazil, over a period of 6 months. The results showed that the nested-PCR was more sensitive (limit of detection: 1.2 PFU/g of tissue) than conventional-PCR and ICC-PCR (limit of detection for both: 1.2 x 10(2)PFU/g of tissue) for detection of Ad5 in oyster extracts. Nested-PCR was able to detect 90% of Ad5 contamination in harvested oyster samples, while conventional-PCR was unable to detect Ad5 in any of the samples. The present work suggests that detection of human adenoviruses can be used as a tool to monitor the presence of human viruses in marine environments where shellfish grow, and that nested-PCR is the method of choice.

  2. Hepatitis E Virus in Wild Boar in Northwest Poland: Sensitivity of Methods of Detection.

    PubMed

    Dorn-In, Samart; Schwaiger, Karin; Twarużek, Magdalena; Grajewski, Jan; Gottschalk, Christoph; Gareis, Manfred

    2017-02-01

    In northwest Poland, 163 blood and 53 fecal samples of wild boars were collected in winter 2012/13 and 2013/14. All blood samples were tested for the presence of hepatitis E virus (HEV) ribonucleic acid (RNA) by two reverse transcription-polymerase chain reaction (RT-PCR) based methods and by anti-HEV IgG enzyme-linked immunosorbent assay (ELISA). About 17.2% of blood samples were seropositive. One-step nested RT-PCR turned out to be too insensitive (11.6% were positive). Therefore a two-step nested RT-PCR was applied where 25.8% of the blood samples were tested positive for HEV RNA. About 50.0% of blood samples positive in ELISA were also positive in two-step nested RT-PCR. The prevalence of HEV RNA in feces was 9.4%. Based on the results of blood (ELISA, PCR) and fecal (PCR) tests, the overall prevalence of HEV in wild boars in northwest Poland was 36.8%. There was no correlation between the ELISA results and the presence of HEV RNA in plasma or in feces. According to the sequencing results of 348 bp PCR products of HEV, there were four different subtypes identified. Reports on the prevalence of HEV in wild boar populations are varying due to different sensitivities of the detection methods. However, this study reveals based on a highly sensitive method that HEV is widely spread in wild boar populations in the northwestern region of Poland and posing a potential risk to the consumer of game meat.

  3. Human papillomavirus detection and typing using a nested-PCR-RFLP assay.

    PubMed

    Coser, Janaina; Boeira, Thaís da Rocha; Fonseca, André Salvador Kazantzi; Ikuta, Nilo; Lunge, Vagner Ricardo

    2011-01-01

    It is clinically important to detect and type human papillomavirus (HPV) in a sensitive and specific manner. Development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-PCR-RFLP) assay to detect and type HPV based on the analysis of L1 gene. Analysis of published DNA sequence of mucosal HPV types to select sequences of new primers. Design of an original nested-PCR assay using the new primers pair selected and classical MY09/11 primers. HPV detection and typing in cervical samples using the nested-PCR-RFLP assay. The nested-PCR-RFLP assay detected and typed HPV in cervical samples. Of the total of 128 clinical samples submitted to simple PCR and nested-PCR for detection of HPV, 37 (28.9%) were positive for the virus by both methods and 25 samples were positive only by nested-PCR (67.5% increase in detection rate compared with single PCR). All HPV positive samples were effectively typed by RFLP assay. The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.

  4. [Roles of histologic examination and polymerase chain reaction in diagnosis of toxoplasmic lymphadenitis].

    PubMed

    Dai, Lin; Huang, Juan; Tang, Yuan; Liao, Dian-ying; Dong, Dan-dan; Xu, Gang; Li, Gan-di

    2010-06-01

    To study the roles of histologic examination and polymerase chain reaction in diagnosis of toxoplasmic lymphadenitis (TL). Forty-six archival cases of histologically diagnosed TL, encountered during the period from April, 1999 to September, 2009 and with the paraffin-embedded lymph node tissue blocks available, were enrolled into the study. The presence of genome fragments of Toxoplasma gondii (T. gondii) was analyzed using semi-nested polymerase chain reaction (PCR). Thirty cases of one or two histopathologic triad of TL as the controls. The positive rate of PCR in TL group was 76.1% (35/46), as compared to 10.0% (3/30) in the control group. The difference was of statistical significance. The sensitivity and specificity of the histologic triad in diagnosing TL was 92.1% (35/38) and 71.1% (27/38), respectively. The predictive value of positive and negative PCR results was 76.1% (35/46) and 90.0% (27/30). respectively. The high specificity but low sensitivity of applying the histologic triad in diagnosing TL cases may be due to the occurrence of atypical histologic pattern. The sensitivity is improved with the use of semi-nested PCR in detecting T. gondii DNA.

  5. Summer bird use of Kansas windbreaks

    USGS Publications Warehouse

    Cable, T.T.; Schroeder, R.L.; Brack, V.; Cook, P.S.

    1992-01-01

    Twenty-four windbreaks in Stafford and Pawnee counties in Kansas were censused for birds during the nesting seasons of 1988-1990. Eighty-nine species of birds were found in the windbreaks with 60 species believed to be nesting. Fifteen species used the windbreaks for feeding or as singing perches but did not nest, and 14 species were spring migrants. Species representing 20 taxonomic families and 15 foraging guilds nested in the windbreaks. Fifteen species were members of the omnivore/ground forager guild. Five species considered forest interior or area sensitive were found nesting in the windbreaks. Windbreak area was important in determining which species were present. Windbreaks provide habitat for a large component of the Great Plains' avifauna, including some species whose populations are decreasing in the region.

  6. Comparison of nested PCR and qPCR for the detection and quantitation of BoHV6 DNA.

    PubMed

    Kubiś, Piotr; Materniak, Magdalena; Kuźmak, Jacek

    2013-12-01

    Nested PCR and qPCR (quantitative PCR) tests based on glycoprotein B (gB) gene were designed for detecting Bovine herpesvirus 6 (BoHV6) in bovine whole blood samples and wild ruminant blood clots (deer and roe-deer). This virus, commonly known as BLHV (bovine lymphotropic herpesvirus) belongs to the Herpesviridae family, subfamily Gammaherpesvirinae and Macavirus genus. DNA isolated from 92 dairy cow blood samples and 69 wild ruminant clots were examined for the presence of BoHV6 using nested PCR and qPCR tests. Viral DNA was detected by using nested PCR in 59 out of 92 bovine blood samples (64.1%), and by qPCR in 68 out of 92 bovine blood samples (73.9%), but none out of 69 DNA samples isolated from wild ruminant blood clots, was positive in both assays. The specificity of nested PCR and qPCR was confirmed by using BoHV1, BoHV4, BoHV6, BFV, BIV, and BLV DNA. The sensitivity of nested PCR and qPCR was determined using a serially 10-fold diluted vector pCR2.1HgB (2 × 10(0)-2 × 10(6)copies/reaction). In this testing, qPCR was more sensitive than the nested PCR, detecting two copies of BoHV6 whilst the limit of detection for nested PCR was 20 copies. In all qPCR assays, the coefficients of determination (R(2)) ranged between 0.990 and 0.999, and the calculated amplification efficiencies (Eff%) within the range of 89.7-106.9. The intra- and inter-assay CV (coefficient of variation) values did not exceed 4%. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Can hedgerow management mitigate the impacts of predation on songbird nest survival?

    PubMed

    Dunn, Jenny C; Gruar, Derek; Stoate, Chris; Szczur, John; Peach, Will J

    2016-12-15

    Nest predators can have significant impacts on songbird reproductive success. These impacts may be amplified by habitat simplification and here we test whether sympathetic management of farmland hedgerows can reduce nest depredation, especially by corvids. We test whether songbirds select nest sites according to structural features of hedgerows (including nest visibility and accessibility), and whether these features influence nest predation risk. Songbirds selected nesting sites affording higher vegetation cover above the nest, increased visibility on the nest-side of the hedgerow and reduced visibility on the far side of the hedge. Nest survival was unrelated to corvid abundance and only weakly related (at the egg stage) to corvid nest proximity. Nest survival at the chick stage was higher where vegetation structure restricted access to corvid-sized predators (averaging 0.78 vs. 0.53), and at nests close to potential vantage points. Overall nest survival was sensitive to hedgerow structure (accessibility) particularly at low exposure to corvid predation, while the overall impact of corvid exposure was dependent on the relationship involving proximity to vantage points. Nest survival over the chick stage was much higher (0.67) in stock-proof, trimmed and mechanically cut hedgerows, (which tended to provide lower side visibility and accessibility) than in recently laid, remnant or leggy hedgerows (0.18). Long-term reductions in the management of British hedgerows may therefore be exposing nesting songbirds to increased predation risk. We recommend regular rotational cutting of hedgerows to maintain a dense woody structure and thereby reduce songbird nest predation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Identification of Aspergillus fumigatus and Related Species by Nested PCR Targeting Ribosomal DNA Internal Transcribed Spacer Regions

    PubMed Central

    Zhao, Jun; Kong, Fanrong; Li, Ruoyu; Wang, Xiaohong; Wan, Zhe; Wang, Duanli

    2001-01-01

    Aspergillus fumigatus is the most common species that causes invasive aspergillosis. In order to identify A. fumigatus, partial ribosomal DNA (rDNA) from two to six strains of five different Aspergillus species was sequenced. By comparing sequence data from GenBank, we designed specific primer pairs targeting rDNA internal transcribed spacer (ITS) regions of A. fumigatus. A nested PCR method for identification of other A. fumigatus-related species was established by using the primers. To evaluate the specificities and sensitivities of those primers, 24 isolates of A. fumigatus and variants, 8 isolates of Aspergillus nidulans, 7 isolates of Aspergillus flavus and variants, 8 isolates of Aspergillus terreus, 9 isolates of Aspergillus niger, 1 isolate each of Aspergillus parasiticus, Aspergillus penicilloides, Aspergillus versicolor, Aspergillus wangduanlii, Aspergillus qizutongii, Aspergillus beijingensis, and Exophiala dermatitidis, 4 isolates of Candida, 4 isolates of bacteria, and human DNA were used. The nested PCR method specifically identified the A. fumigatus isolates and closely related species and showed a high degree of sensitivity. Additionally, four A. fumigatus strains that were recently isolated from our clinic were correctly identified by this method. Our results demonstrate that these primers are useful for the identification of A. fumigatus and closely related species in culture and suggest further studies for the identification of Aspergillus fumigatus species in clinical specimens. PMID:11376067

  9. Occurrence of killer yeasts in leaf-cutting ant nests.

    PubMed

    Carreiro, S C; Pagnocca, F C; Bacci, M; Bueno, O C; Hebling, M J A; Middelhoven, W J

    2002-01-01

    Killer activity was screened in 99 yeast strains isolated from the nests of the leaf-cutting ant Atta sexdens against 6 standard sensitive strains, as well as against each other. Among this yeast community killer activity was widespread since 77 strains (78%) were able to kill or inhibit the growth of at least one standard strain or nest strain. Toxin production was observed in representatives of all the studied genera including Aureobasidium, Rhodotorula, Tremella and Trichosporon, whose killer activity has not yet been described.

  10. Aquatic Nitrate Retention at River Network Scales Across Flow Conditions Determined Using Nested In Situ Sensors

    NASA Astrophysics Data System (ADS)

    Wollheim, W. M.; Mulukutla, G. K.; Cook, C.; Carey, R. O.

    2017-11-01

    Nonpoint pollution sources are strongly influenced by hydrology and are therefore sensitive to climate variability. Some pollutants entering aquatic ecosystems, e.g., nitrate, can be mitigated by in-stream processes during transport through river networks. Whole river network nitrate retention is difficult to quantify with observations. High frequency, in situ nitrate sensors, deployed in nested locations within a single watershed, can improve estimates of both nonpoint inputs and aquatic retention at river network scales. We deployed a nested sensor network and associated sampling in the urbanizing Oyster River watershed in coastal New Hampshire, USA, to quantify storm event-scale loading and retention at network scales. An end member analysis used the relative behavior of reactive nitrate and conservative chloride to infer river network fate of nitrate. In the headwater catchments, nitrate and chloride concentrations are both increasingly diluted with increasing storm size. At the mouth of the watershed, chloride is also diluted, but nitrate tended to increase. The end member analysis suggests that this pattern is the result of high retention during small storms (51-78%) that declines to zero during large storms. Although high frequency nitrate sensors did not alter estimates of fluxes over seasonal time periods compared to less frequent grab sampling, they provide the ability to estimate nitrate flux versus storm size at event scales that is critical for such analyses. Nested sensor networks can improve understanding of the controls of both loading and network scale retention, and therefore also improve management of nonpoint source pollution.

  11. Comparative effects of Rauwolfia vomitoria and chlorpromazine on social behaviour and pain

    PubMed Central

    Bisong, Sunday; Brown, Richard; Osim, Eme

    2011-01-01

    Background: Rauwolfia vomitoria has been used in Nigeria to manage psychiatric disorders despite orthodox medicine. Aims: This research was therefore aimed at comparing the effects of R. vomitoria, chlorpromazine and reserpine on social behaviour and pain in mice. Materials and Methods: Ninety male CD-1 mice (32 – 38g body weight) were grouped into 3 with 5 subgroups (n=6) each. Mice were given chlorpromazine (0.0, 0.25, 1.0, 2.0, 4.0 mg/kg i.p.), 30 minutes before testing and R. vomitoria (0.0, 0.25, 1.0, 2.0, 4.0 mg/kg, i.p.) and reserpine (0.0, 0.1, 0.4, 0.8, 1.6 mg/kg, i.p) 24 hours before testing. Nesting score assessed social behaviour while the tail flick and hot plate analgesiometers assessed pain. Results: Chlorpromazine dose-dependently decreased nesting score (F4,25 = 5.5660; p< 0.01), indicating decreased social behaviour (social loss) in the mice. Although R. vomitoria did not affect nesting score, reserpine decreased the nesting score (social loss). In the pain test, chlorpromazine did not alter tail flick latency but decreased hind paw lick latency in the hot plate at 2.0 and 4.0 mg/kg (p< 0.01), indicating increased pain sensitivity at these doses which may indirectly increase social withdrawal and thus aggravating depression. R. vomitoria however, increased tail flick and hind paw lick latencies in the hot plate test (p< 0.05) indicating decreased pain sensitivity. Reserpine, like R. vomitoria, increased latency of hind paw lick in the hot plate. Conclusion: R. vomitoria has a high potential as an antipsychotic and may have advantage over chlorpromazine; it is not necessary to isolate active components from this herb. PMID:22540065

  12. Ovarian Brenner tumour: a morphologic and immunohistochemical analysis suggesting an origin from fallopian tube epithelium.

    PubMed

    Kuhn, Elisabetta; Ayhan, Ayse; Shih, Ie-Ming; Seidman, Jeffrey D; Kurman, Robert J

    2013-12-01

    Brenner tumours (BTs), like other epithelial ovarian tumours, are thought to develop from the ovarian surface epithelium. We hypothesised that BTs arise from transitional metaplasia near the tuboperitoneal junction which, when embedded in the ovary as Walthard cell nests, may progress to BTs. The aim of this study was to validate this hypothesis by a morphologic and immunohistochemical (IHC) analysis. The IHC analysis revealed that fallopian tube secretory cells, transitional metaplasia, Walthard cell nests and the epithelial component of BTs shared a similar IHC profile, consistently expressing AKR1C3 (an enzyme involved in androgen biosynthesis) and androgen receptor, but not calretinin. The tumour stromal cells that immediately surrounded the epithelial nests showed strong expression of calretinin, inhibin and steroidogenic factor 1 (markers of steroidogenic cells) in the majority of BTs. Using a highly sensitive immunofluorescent staining method, we detected small groups of cilia in transitional metaplasia and Walthard cell nests, multifocal stretches of cilia and/or ciliated vacuoles in benign BTs and well-developed cilia in atypical proliferative BTs. Our findings suggest a tubal origin of BTs through transitional metaplasia and Walthard cell nests, based on their anatomic proximity, similar IHC profile and the presence of cilia. In addition, we hypothesise a role of androgenic stimulation in the pathogenesis of BT, based on the IHC staining pattern of calretinin, inhibin and steroidogenic factor 1 expressed in the luteinised stromal cells surrounding the epithelial nests of the tumours, and AKR1C3 and androgen receptor expressed in both the epithelial and stromal components. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Integrated and isolated impact of high-performance work practices on employee health and well-being: A comparative study.

    PubMed

    Ogbonnaya, Chidiebere; Daniels, Kevin; Connolly, Sara; van Veldhoven, Marc

    2017-01-01

    We investigate the positive relationships between high-performance work practices (HPWP) and employee health and well-being and examine the conflicting assumption that high work intensification arising from HPWP might offset these positive relationships. We present new insights on whether the combined use (or integrated effects) of HPWP has greater explanatory power on employee health, well-being, and work intensification compared to their isolated or independent effects. We use data from the 2004 British Workplace Employment Relations Survey (22,451 employees nested within 1,733 workplaces) and the 2010 British National Health Service Staff survey (164,916 employees nested within 386 workplaces). The results show that HPWP have positive combined effects in both contexts, and work intensification has a mediating role in some of the linkages investigated. The results also indicate that the combined use of HPWP may be sensitive to particular organizational settings, and may operate in some sectors but not in others. (PsycINFO Database Record (c) 2016 APA, all rights reserved).

  14. Spatiotemporal patterns of duck nest density and predation risk: a multi-scale analysis of 18 years and more than 10,000 nests

    USGS Publications Warehouse

    Ringelman, Kevin M.; Eadie, John M.; Ackerman, Joshua T.; Sih, Andrew; Loughman, Daniel L.; Yarris, Gregory S.; Oldenburger, Shaun L.; McLandress, M. Robert

    2017-01-01

    Many avian species are behaviorally-plastic in selecting nest sites, and may shift to new locations or habitats following an unsuccessful breeding attempt. If there is predictable spatial variation in predation risk, the process of many individuals using prior experience to adaptively change nest sites may scale up to create shifting patterns of nest density at a population level. We used 18 years of waterfowl nesting data to assess whether there were areas of consistently high or low predation risk, and whether low-risk areas increased, and high-risk areas decreased in nest density the following year. We created kernel density maps of successful and unsuccessful nests in consecutive years and found no correlation in predation risk and no evidence for adaptive shifts, although nest density was correlated between years. We also examined between-year correlations in nest density and nest success at three smaller spatial scales: individual nesting fields (10–28 ha), 16-ha grid cells and 4-ha grid cells. Here, results were similar across all scales: we found no evidence for year-to-year correlation in nest success but found strong evidence that nest density was correlated between years, and areas of high nest success increased in nest density the following year. Prior research in this system has demonstrated that areas of high nest density have higher nest success, and taken together, our results suggest that ducks may adaptively select nest sites based on the local density of conspecifics, rather than the physical location of last year's nest. In unpredictable environments, current cues, such as the presence of active conspecific nests, may be especially useful in selecting nest sites. The cues birds use to select breeding locations and successfully avoid predators deserve continued attention, especially in systems of conservation concern.

  15. Differential Detection of Enterovirus and Herpes Simplex Virus in Cerebrospinal Fluid by Real-Time RT-PCR.

    PubMed

    Sarquiz-Martínez, Brenda; González-Bonilla, César R; Santacruz-Tinoco, Clara Esperanza; Muñoz-Medina, José E; Pardavé-Alejandre, Héctor D; Barbosa-Cabrera, Elizabeth; Ramírez-González, José Ernesto; Díaz-Quiñonez, José Alberto

    2017-01-01

    Enterovirus (EV) and herpes simplex virus 1 and 2 (HSV1 and HSV2) are the main etiologic agents of central nervous system infections. Early laboratory confirmation of these infections is performed by viral culture of the cerebrospinal fluid (CSF), or the detection of specific antibodies in serum (e.g., HSV). The sensitivity of viral culture ranges from 65 to 75%, with a recovery time varying from 3 to 10 days. Serological tests are faster and easy to carry out, but they exhibit cross-reactivity between HSV1 and HSV2. Although molecular techniques are more sensitive (sensitivity >95%), they are more expensive and highly susceptible to cross-contamination. A real-time RT-PCR for the detection of EV, HSV1, and HSV2 was compared with end-point nested PCR. We tested 87 CSF samples of patients with a clinical diagnosis of viral meningitis or encephalitis. Fourteen samples were found to be positive by RT-PCR, but only 8 were positive by end-point PCR. The RT-PCR showed a specificity range of 94-100%, the negative predictive value was 100%, and the positive predictive value was 62, 100, and 28% for HSV1, HSV2, and EV, respectively. Real-time RT-PCR detected EV, HSV1, and HSV2 with a higher sensitivity and specificity than end-point nested RT-PCR. © 2017 S. Karger AG, Basel.

  16. Nested-PCR real time as alternative molecular tool for detection of Borrelia burgdorferi compared to the classical serological diagnosis of the blood.

    PubMed

    Sroka-Oleksiak, Agnieszka; Ufir, Krzysztof; Salamon, Dominika; Bulanda, Malgorzata; Gosiewski, Tomasz

    Lyme disease, caused by Borrelia burgdorferi, is a multisystem disease that often makes difficulties to recognize caused by their genetic heterogenity. Currently, the gold standard for the detection of Lyme disease (LD) is serologic diagnostics based mainly on tests: ELISA and Western blot (WB). These methods, however, are subject to consider- able defect, especially in the initial phase of infection due to the occurrence of so-called serological window period and low specificity. For this reason, they might be replaced by molecular methods, for example polymerase chain reaction (PCR), which should be more sensitivity and specificity. In the present study we attempt to optimize the PCR reaction conditions and enhance existing test sensitivity by applying the equivalent of real time PCR - nested PCR for detection B. burgdorferi DNA in the patient's blood. The study involved 94 blood samples of patients with suspected LD. From each sample, 1.5 ml of blood was used for the isolation of bacterial DNA and PCR real time am- plification and its equivalent, in nested version. The remaining part earmarked for serologi- cal testing. Optimization of the reaction conditions made experimentally, using gradient of the temperature and gradient of the magnesium ions concentration for reaction real time in nested-PCR and PCR version. The results show that the nested-PCR real time, has a much higher sensitivity 45 (47.8%) of positive results for the detection of B. burgdorferi compared to the single- variety, without a preceding pre-amplification 2 (2.1%). Serological methods allowed the detection of infection in 41 (43.6%) samples. These results support of the nested PCR method as a better molecular tool for the detection of B. burgdorferi infection than classical PCR real time reaction. The nested-PCR real time method may be considered as a complement to ELISA and WB mainly in the early stages of infection, when in the blood circulating B. burgdorferi cells. By contrast, the results of serological and molecular tests should always be carried out tak- ing into account the patient's clinical status.

  17. Clinical Usefulness of a One-Tube Nested Reverse Transcription Quantitative Polymerase Chain Reaction Assay for Evaluating Human Epidermal Growth Factor Receptor 2 mRNA Overexpression in Formalin-Fixed and Paraffin-Embedded Breast Cancer Tissue Samples.

    PubMed

    Wang, Hye-Young; Ahn, Sungwoo; Park, Sunyoung; Kim, SeungIl; Lee, Hyeyoung

    2017-01-01

    Currently, the two main methods used to analyze human epidermal growth factor receptor 2 (HER2) amplification or overexpression have a limited accuracy and high costs. These limitations can be overcome by the development of complementary quantitative methods. In this study, we analyzed HER2 mRNA expression in clinical formalin-fixed and paraffin-embedded (FFPE) samples using a one-tube nested reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay. We measured expression relative to 3 reference genes and compared the results to those obtained by conventional immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) assays with 226 FFPE breast cancer tissue samples. The one-tube nested RT-qPCR assay proved to be highly sensitive and specific based on comparisons with IHC (96.9 and 97.7%, respectively) and FISH (92.4 and 92.9%, respectively) obtained with the validation set. Comparisons with clinicopathological data revealed significant associations between HER2 overexpression and TNM stage (p < 0.01), histological type (p < 0.01), ER status (p < 0.001), PR status (p < 0.05), HER2 status (p < 0.001), and molecular subtypes (p < 0.001). Based on these findings, our one-tube nested RT-qPCR assay is a potentially useful and complementary screening tool for the detection of HER2 mRNA overexpression. © 2016 S. Karger AG, Basel.

  18. A simple Bird Sensitivity to Oil Index as a management tool in coastal and marine areas subject to oil spills when few biological information is available.

    PubMed

    Romero, A F; Oliveira, M; Abessa, D M S

    2018-03-01

    This study sought to develop a simple index for ranking birds' environmental sensitivity to oil in which birds are used as biological indicators. The study area consisted of both the Santos Estuarine System (SES), and the Laje de Santos Marine State Park (LSMSP), located in Southeastern Brazil. Information on the bird species and their feeding and nesting behaviors were obtained from the literature and were the basis of the sensitivity index created. The SES had a higher number of species, but only about 30% were found to be highly sensitive. The LSMSP presented a much lower number of species, but all of them were considered to be highly sensitive to oil. Due to its simplicity, this index can be employed worldwide as a decision-making tool that may be integrated into other management tools, particularly when robust information on the biology of birds is lacking. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Influence of an extreme high water event on survival, reproduction, and distribution of snail kites in Florida, USA

    USGS Publications Warehouse

    Bennetts, R.E.; Kitchens, W.M.; Dreitz, V.J.

    2002-01-01

    Hydrology frequently has been reported as the environmental variable having the greatest influence on Florida snail kite (Rostrhamus sociabilis) populations. Although drought has received the most attention, high-water conditions also have been reported to affect kites. Years of high water generally have been reported to be favorable for nesting, although prolonged high water may be detrimental to sustaining suitable habitat. During 1994 and 1995, southern Florida experienced an extreme high water event. This event enabled us to compare survival, nesting success, number of young per successful nest, and spatial distribution of nesting before, during, and after the event. We found no evidence of an effect (either negative or positive) on survival of adult kites. In contrast, juvenile kites experienced the highest survival during the event, although our data suggest greater annual variability than can be explained by the event alone. We found no evidence of an effect of the high water event on nest success or number of young per successful nest. Nest success was highest during the event in the southern portion of the range but was quite similar to other years, both before and after the event. Our data do indicate a substantial shift in the spatial distribution of nesting birds. During the event, nesting activity shifted to higher elevations (i.e., shallower water) in the major nesting areas of the Everglades region. Nesting also occurred in Big Cypress National Preserve during the event, which is typically too dry to support nesting kites. Thus, our data indicate a potential short-term benefit of increased juvenile survival and an expansion of nesting habitat. However, the deterioration of habitat quality from prolonged high water precludes any recommendation for such conditions to be maintained for extended periods. ?? 2002, The Society of Wetland Scientists.

  20. A comparison between the efficiency of the Xpert MTB/RIF assay and nested PCR in identifying Mycobacterium tuberculosis during routine clinical practice.

    PubMed

    Kim, Cheol-Hong; Woo, Heungjeong; Hyun, In Gyu; Kim, Changhwan; Choi, Jeong-Hee; Jang, Seung-Hun; Park, Sang Myeon; Kim, Dong-Gyu; Lee, Myung Goo; Jung, Ki-Suck; Hyun, Jeongwon; Kim, Hyun Soo

    2014-06-01

    Polymerase chain reaction (PCR) for the detection of Mycobacterium tuberculosis (MTB) is more sensitive, specific, and rapid than the conventional methods of acid-fast bacilli (AFB) smear and culture. The aim of this study was to determine if the Xpert MTB/rifampicin (RIF) assay had additional advantages over nested PCR for the detection of MTB in a geographical area with intermediate tuberculosis (TB) incidence. Between February and December 2013, the Xpert MTB/RIF assay and MTB nested PCR, as well as AFB smear and culture, were simultaneously performed on 198 clinical samples (160 pulmonary and 38 non-pulmonary specimens) collected from 171 patients hospitalized at Hallym University Medical Center for possible TB. The accuracy of the diagnosis of MTB culture-positive TB and the turnaround time of reporting laboratory results were calculated and compared. Rifampin resistance by the Xpert MTB/RIF assay was reviewed with that of conventional drug susceptibility testing (DST). The sensitivity, specificity, and positive and negative predictive values of the Xpert MTB/RIF assay and MTB nested PCR for diagnosis of MTB culture-positive pulmonary TB were 86.1% vs. 69.4% (P=0.1563), 97.8% vs. 94.1% (P=0.2173), 91.2% vs. 75.8% (P=0.1695), and 96.4% vs. 92.0% (P=0.2032), respectively. The median turnaround times of the Xpert MTB/RIF assay and MTB nested PCR were 0 [0-4] days and 4 [1-11] days, respectively (P<0.001). Two cases of rifampin resistance, as determined by the Xpert MTB/RIF assay, were found to be multi-drug resistant (MDR) pulmonary TB by DST. The Xpert MTB/RIF assay seemed to be sensitive, specific, and comparable to nested PCR for identifying MTB among clinically suspected TB patients, and the assay can be valuable in giving a timely identification of resistance to rifampin.

  1. Highly Sensitive Detection of Low-Abundance White Spot Syndrome Virus by a Pre-Amplification PCR Method.

    PubMed

    Pan, Xiaoming; Zhang, Yanfang; Sha, Xuejiao; Wang, Jing; Li, Jing; Dong, Ping; Liang, Xingguo

    2017-03-28

    White spot syndrome virus (WSSV) is a major threat to the shrimp farming industry and so far there is no effective therapy for it, and thus early diagnostic of WSSV is of great importance. However, at the early stage of infection, the extremely low-abundance of WSSV DNA challenges the detection sensitivity and accuracy of PCR. To effectively detect low-abundance WSSV, here we developed a pre-amplification PCR (pre-amp PCR) method to amplify trace amounts of WSSV DNA from massive background genomic DNA. Combining with normal specific PCR, 10 copies of target WSSV genes were detected from ~10 10 magnitude of backgrounds. In particular, multiple target genes were able to be balanced amplified with similar efficiency due to the usage of the universal primer. The efficiency of the pre-amp PCR was validated by nested-PCR and quantitative PCR, and pre-amp PCR showed higher efficiency than nested-PCR when multiple targets were detected. The developed method is particularly suitable for the super early diagnosis of WSSV, and has potential to be applied in other low-abundance sample detection cases.

  2. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR.

    PubMed

    Araújo, Cristina P; Osório, Ana Luiza A R; Jorge, Klaudia S G; Ramos, Carlos A N; Souza Filho, Antonio F; Vidal, Carlos E S; Vargas, Agueda P C; Roxo, Eliana; Rocha, Adalgiza S; Suffys, Philip N; Fonseca, Antônio A; Silva, Marcio R; Barbosa Neto, José D; Cerqueira, Valíria D; Araújo, Flábio R

    2014-01-01

    Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

  3. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

    PubMed Central

    Araújo, Cristina P.; Osório, Ana Luiza A.R.; Jorge, Klaudia S.G.; Ramos, Carlos A.N.; Souza Filho, Antonio F.; Vidal, Carlos E.S.; Vargas, Agueda P.C.; Roxo, Eliana; Rocha, Adalgiza S.; Suffys, Philip N.; Fonseca, Antônio A.; Silva, Marcio R.; Barbosa Neto, José D.; Cerqueira, Valíria D.; Araújo, Flábio R.

    2014-01-01

    Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis. PMID:25242951

  4. A Nested Phosphorus and Proton Coil Array for Brain Magnetic Resonance Imaging and Spectroscopy

    PubMed Central

    Brown, Ryan; Lakshmanan, Karthik; Madelin, Guillaume; Parasoglou, Prodromos

    2015-01-01

    A dual-nuclei radiofrequency coil array was constructed for phosphorus and proton magnetic resonance imaging and spectroscopy of the human brain at 7 Tesla. An eight-channel transceive degenerate birdcage phosphorus module was implemented to provide whole-brain coverage and significant sensitivity improvement over a standard dual-tuned loop coil. A nested eight-channel proton module provided adequate sensitivity for anatomical localization without substantially sacrificing performance on the phosphorus module. The developed array enabled phosphorus spectroscopy, a saturation transfer technique to calculate the global creatine kinase forward reaction rate, and single-metabolite whole-brain imaging with 1.4 cm nominal isotropic resolution in 15 min (2.3 cm actual resolution), while additionally enabling 1 mm isotropic proton imaging. This study demonstrates that a multi-channel array can be utilized for phosphorus and proton applications with improved coverage and/or sensitivity over traditional single-channel coils. The efficient multi-channel coil array, time-efficient pulse sequences, and the enhanced signal strength available at ultra-high fields can be combined to allow volumetric assessment of the brain and could provide new insights into the underlying energy metabolism impairment in several neurodegenerative conditions, such as Alzheimer’s and Parkinson’s diseases, as well as mental disorders such as schizophrenia. PMID:26375209

  5. A nested phosphorus and proton coil array for brain magnetic resonance imaging and spectroscopy.

    PubMed

    Brown, Ryan; Lakshmanan, Karthik; Madelin, Guillaume; Parasoglou, Prodromos

    2016-01-01

    A dual-nuclei radiofrequency coil array was constructed for phosphorus and proton magnetic resonance imaging and spectroscopy of the human brain at 7T. An eight-channel transceive degenerate birdcage phosphorus module was implemented to provide whole-brain coverage and significant sensitivity improvement over a standard dual-tuned loop coil. A nested eight-channel proton module provided adequate sensitivity for anatomical localization without substantially sacrificing performance on the phosphorus module. The developed array enabled phosphorus spectroscopy, a saturation transfer technique to calculate the global creatine kinase forward reaction rate, and single-metabolite whole-brain imaging with 1.4cm nominal isotropic resolution in 15min (2.3cm actual resolution), while additionally enabling 1mm isotropic proton imaging. This study demonstrates that a multi-channel array can be utilized for phosphorus and proton applications with improved coverage and/or sensitivity over traditional single-channel coils. The efficient multi-channel coil array, time-efficient pulse sequences, and the enhanced signal strength available at ultra-high fields can be combined to allow volumetric assessment of the brain and could provide new insights into the underlying energy metabolism impairment in several neurodegenerative conditions, such as Alzheimer's and Parkinson's diseases, as well as mental disorders such as schizophrenia. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Note: A sample holder design for sensitive magnetic measurements at high temperatures in a magnetic properties measurement system

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Arauzo, A.; Guerrero, E.; Urtizberea, A.

    2012-06-15

    A sample holder design for high temperature measurements in a commercial MPMS SQUID magnetometer from Quantum Design is presented. It fulfills the requirements for the simultaneous use of the oven and reciprocating sample option (RSO) options, thus allowing sensitive magnetic measurements up to 800 K. Alternating current susceptibility can also be measured, since the holder does not induce any phase shift relative to the ac driven field. It is easily fabricated by twisting Constantan Copyright-Sign wires into a braid nesting the sample inside. This design ensures that the sample be placed tightly into a tough holder with its orientation fixed,more » and prevents any sample displacement during the fast movements of the RSO transport, up to high temperatures.« less

  7. Enhanced detection of tuberculous mycobacteria in animal tissues using a semi-nested probe-based real-time PCR.

    PubMed

    Costa, Pedro; Ferreira, Ana S; Amaro, Ana; Albuquerque, Teresa; Botelho, Ana; Couto, Isabel; Cunha, Mónica V; Viveiros, Miguel; Inácio, João

    2013-01-01

    Bovine tuberculosis has been tackled for decades by costly eradication programs in most developed countries, involving the laboratory testing of tissue samples from allegedly infected animals for detection of Mycobacterium tuberculosis complex (MTC) members, namely Mycobacterium bovis. Definitive diagnosis is usually achieved by bacteriological culture, which may take up to 6-12 weeks, during which the suspect animal carcass and herd are under sanitary arrest. In this work, a user-friendly DNA extraction protocol adapted for tissues was coupled with an IS6110-targeted semi-nested duplex real-time PCR assay to enhance the direct detection of MTC bacteria in animal specimens, reducing the time to achieve a diagnosis and, thus, potentially limiting the herd restriction period. The duplex use of a novel β-actin gene targeted probe, with complementary targets in most mammals, allowed the assessment of amplification inhibitors in the tissue samples. The assay was evaluated with a group of 128 fresh tissue specimens collected from bovines, wild boars, deer and foxes. Mycobacterium bovis was cultured from 57 of these samples. Overall, the full test performance corresponds to a diagnostic sensitivity and specificity of 98.2% (CIP95% 89.4-99.9%) and 88.7% (CIP95% 78.5-94.7%), respectively. An observed kappa coefficient was estimated in 0.859 (CI P95% 0.771-0.948) for the overall agreement between the semi-nested PCR assay and the bacteriological culture. Considering only bovine samples (n = 69), the diagnostic sensitivity and specificity were estimated in 100% (CIP95% 84.0-100%) and 97.7% (CIP95% 86.2-99.9%), respectively. Eight negative culture samples exhibiting TB-like lesions were detected by the semi-nested real-time PCR, thus emphasizing the increased potential of this molecular approach to detect MTC-infected animal tissues. This novel IS6110-targeted assay allows the fast detection of tuberculous mycobacteria in animal specimens with very high sensitivity and specificity, being amenable and cost effective for use in the routine veterinary diagnostic laboratory with further automation possibilities.

  8. Loop mediated isothermal amplification (LAMP) accurately detects malaria DNA from filter paper blood samples of low density parasitaemias.

    PubMed

    Aydin-Schmidt, Berit; Xu, Weiping; González, Iveth J; Polley, Spencer D; Bell, David; Shakely, Delér; Msellem, Mwinyi I; Björkman, Anders; Mårtensson, Andreas

    2014-01-01

    Loop mediated isothermal amplification (LAMP) provides an opportunity for improved, field-friendly detection of malaria infections in endemic areas. However data on the diagnostic accuracy of LAMP for active case detection, particularly low-density parasitaemias, are lacking. We therefore evaluated the performance of a new LAMP kit compared with PCR using DNA from filter paper blood spots. Samples from 865 fever patients and 465 asymptomatic individuals collected in Zanzibar were analysed for Pan (all species) and Pf (P. falciparum) DNA with the Loopamp MALARIA Pan/Pf kit. Samples were amplified at 65°C for 40 minutes in a real-time turbidimeter and results were compared with nested PCR. Samples with discordant results between LAMP and nested PCR were analysed with real-time PCR. The real-time PCR corrected nested PCR result was defined as gold standard. Among the 117 (13.5%) PCR detected P. falciparum infections from fever patients (mean parasite density 7491/µL, range 6-782,400) 115, 115 and 111 were positive by Pan-LAMP, Pf-LAMP and nested PCR, respectively. The sensitivities were 98.3% (95%CI 94-99.8) for both Pan and Pf-LAMP. Among the 54 (11.6%) PCR positive samples from asymptomatic individuals (mean parasite density 10/µL, range 0-4972) Pf-LAMP had a sensitivity of 92.7% (95%CI 80.1-98.5) for detection of the 41 P. falciparum infections. Pan-LAMP had sensitivities of 97% (95%CI 84.2-99.9) and 76.9% (95%CI 46.2-95) for detection of P. falciparum and P. malariae, respectively. The specificities for both Pan and Pf-LAMP were 100% (95%CI 99.1-100) in both study groups. Both components of the Loopamp MALARIA Pan/Pf detection kit revealed high diagnostic accuracy for parasite detection among fever patients and importantly also among asymptomatic individuals of low parasite densities from minute blood volumes preserved on filter paper. These data support LAMPs potential role for improved detection of low-density malaria infections in pre-elimination settings.

  9. Loop Mediated Isothermal Amplification (LAMP) Accurately Detects Malaria DNA from Filter Paper Blood Samples of Low Density Parasitaemias

    PubMed Central

    González, Iveth J.; Polley, Spencer D.; Bell, David; Shakely, Delér; Msellem, Mwinyi I.; Björkman, Anders; Mårtensson, Andreas

    2014-01-01

    Background Loop mediated isothermal amplification (LAMP) provides an opportunity for improved, field-friendly detection of malaria infections in endemic areas. However data on the diagnostic accuracy of LAMP for active case detection, particularly low-density parasitaemias, are lacking. We therefore evaluated the performance of a new LAMP kit compared with PCR using DNA from filter paper blood spots. Methods and Findings Samples from 865 fever patients and 465 asymptomatic individuals collected in Zanzibar were analysed for Pan (all species) and Pf (P. falciparum) DNA with the Loopamp MALARIA Pan/Pf kit. Samples were amplified at 65°C for 40 minutes in a real-time turbidimeter and results were compared with nested PCR. Samples with discordant results between LAMP and nested PCR were analysed with real-time PCR. The real-time PCR corrected nested PCR result was defined as gold standard. Among the 117 (13.5%) PCR detected P. falciparum infections from fever patients (mean parasite density 7491/µL, range 6–782,400) 115, 115 and 111 were positive by Pan-LAMP, Pf-LAMP and nested PCR, respectively. The sensitivities were 98.3% (95%CI 94–99.8) for both Pan and Pf-LAMP. Among the 54 (11.6%) PCR positive samples from asymptomatic individuals (mean parasite density 10/µL, range 0–4972) Pf-LAMP had a sensitivity of 92.7% (95%CI 80.1–98.5) for detection of the 41 P. falciparum infections. Pan-LAMP had sensitivities of 97% (95%CI 84.2–99.9) and 76.9% (95%CI 46.2–95) for detection of P. falciparum and P. malariae, respectively. The specificities for both Pan and Pf-LAMP were 100% (95%CI 99.1–100) in both study groups. Conclusion Both components of the Loopamp MALARIA Pan/Pf detection kit revealed high diagnostic accuracy for parasite detection among fever patients and importantly also among asymptomatic individuals of low parasite densities from minute blood volumes preserved on filter paper. These data support LAMPs potential role for improved detection of low-density malaria infections in pre-elimination settings. PMID:25105591

  10. PCR diagnostics underestimate the prevalence of avian malaria (Plasmodium relictum) in experimentally-infected passerines

    USGS Publications Warehouse

    Jarvi, Susan I.; Schultz, Jeffrey J.; Atkinson, Carter T.

    2002-01-01

    Several polymerase chain reaction (PCR)-based methods have recently been developed for diagnosing malarial infections in both birds and reptiles, but a critical evaluation of their sensitivity in experimentally-infected hosts has not been done. This study compares the sensitivity of several PCR-based methods for diagnosing avian malaria (Plasmodium relictum) in captive Hawaiian honeycreepers using microscopy and a recently developed immunoblotting technique. Sequential blood samples were collected over periods of up to 4.4 yr after experimental infection and rechallenge to determine both the duration and detectability of chronic infections. Two new nested PCR approaches for detecting circulating parasites based on P. relictum 18S rRNA genes and the thrombospondin-related anonymous protein (TRAP) gene are described. The blood smear and the PCR tests were less sensitive than serological methods for detecting chronic malarial infections. Individually, none of the diagnostic methods was 100% accurate in detecting subpatent infections, although serological methods were significantly more sensitive (97%) than either nested PCR (61–84%) or microscopy (27%). Circulating parasites in chronically infected birds either disappear completely from circulation or to drop to intensities below detectability by nested PCR. Thus, the use of PCR as a sole means of detection of circulating parasites may significantly underestimate true prevalence.

  11. Visual Navigation in Nocturnal Insects.

    PubMed

    Warrant, Eric; Dacke, Marie

    2016-05-01

    Despite their tiny eyes and brains, nocturnal insects have evolved a remarkable capacity to visually navigate at night. Whereas some use moonlight or the stars as celestial compass cues to maintain a straight-line course, others use visual landmarks to navigate to and from their nest. These impressive abilities rely on highly sensitive compound eyes and specialized visual processing strategies in the brain. ©2016 Int. Union Physiol. Sci./Am. Physiol. Soc.

  12. H. pylori seroprevalence and risk of diabetes: An ancillary case-control study nested in the diabetes prevention program.

    PubMed

    Alzahrani, Saud; Nelson, Jason; Moss, Steven F; Paulus, Jessica K; Knowler, William C; Pittas, Anastassios G

    2017-10-01

    To determine the association between H. pylori infection and risk of incident diabetes in adults at high risk for diabetes who participated in the Diabetes Prevention Program (DPP) study. In a nested case-control study conducted among 421 adults with newly diagnosed diabetes and 421 matched controls, we examined the association between serological status of H. pylori at baseline and risk of incident diabetes over a mean follow-up period of 2.6years. Using data from the baseline visit of the DPP, we also examined the cross-sectional association between presence of H. pylori antibodies and insulin sensitivity, insulin secretion and the disposition index-like measure after a 75-g oral glucose tolerance test (OGTT). At baseline, H. pylori antibodies were present in 40% of participants who developed diabetes and 39% of controls. After adjusting for matching factors, there was no association between exposure to H. pylori and incident diabetes (odds ratio [OR] of 1.04 (95% CI, 0.77 to 1.40). In cross-sectional analyses, H. pylori status was not significantly associated with insulin sensitivity and disposition index-like measure from OGTT. In adults at high risk for diabetes, H. pylori seropositivity was not associated with risk of developing diabetes. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Highly controlled nest homeostasis of honey bees helps deactivate phenolics in nectar

    NASA Astrophysics Data System (ADS)

    Liu, Fanglin; He, Jianzhong; Fu, Wenjun

    2005-06-01

    Honey bees have a highly developed nest homeostasis, for example, maintaining low CO2 levels and stable nest temperatures at 35°C.We investigate the role of nest homeostasis in deactivating phenolic compounds present in the nectar of Aloe littoralis. We show that the phenolic content in nectar was reduced (from 0.65% to 0.49%) after nectar was incubated in a nest of Apis cerana, and that it was reduced still more (from 0.65% to 0.37%) if nectar was mixed with hypopharyngeal gland proteins (HGP) of worker bees before being placed inside a nest. HGP had little effect on samples outside a nest, indicating that nest conditions are necessary for HGP to deactivate phenolics in nectar. Consequently, the highly controlled nest homeostasis of honey bees facilitates direct deactivation of phenolics in nectar, and plays a role in the action of HGP as well.

  14. Model of the best-of-N nest-site selection process in honeybees.

    PubMed

    Reina, Andreagiovanni; Marshall, James A R; Trianni, Vito; Bose, Thomas

    2017-05-01

    The ability of a honeybee swarm to select the best nest site plays a fundamental role in determining the future colony's fitness. To date, the nest-site selection process has mostly been modeled and theoretically analyzed for the case of binary decisions. However, when the number of alternative nests is larger than two, the decision-process dynamics qualitatively change. In this work, we extend previous analyses of a value-sensitive decision-making mechanism to a decision process among N nests. First, we present the decision-making dynamics in the symmetric case of N equal-quality nests. Then, we generalize our findings to a best-of-N decision scenario with one superior nest and N-1 inferior nests, previously studied empirically in bees and ants. Whereas previous binary models highlighted the crucial role of inhibitory stop-signaling, the key parameter in our new analysis is the relative time invested by swarm members in individual discovery and in signaling behaviors. Our new analysis reveals conflicting pressures on this ratio in symmetric and best-of-N decisions, which could be solved through a time-dependent signaling strategy. Additionally, our analysis suggests how ecological factors determining the density of suitable nest sites may have led to selective pressures for an optimal stable signaling ratio.

  15. Model of the best-of-N nest-site selection process in honeybees

    NASA Astrophysics Data System (ADS)

    Reina, Andreagiovanni; Marshall, James A. R.; Trianni, Vito; Bose, Thomas

    2017-05-01

    The ability of a honeybee swarm to select the best nest site plays a fundamental role in determining the future colony's fitness. To date, the nest-site selection process has mostly been modeled and theoretically analyzed for the case of binary decisions. However, when the number of alternative nests is larger than two, the decision-process dynamics qualitatively change. In this work, we extend previous analyses of a value-sensitive decision-making mechanism to a decision process among N nests. First, we present the decision-making dynamics in the symmetric case of N equal-quality nests. Then, we generalize our findings to a best-of-N decision scenario with one superior nest and N -1 inferior nests, previously studied empirically in bees and ants. Whereas previous binary models highlighted the crucial role of inhibitory stop-signaling, the key parameter in our new analysis is the relative time invested by swarm members in individual discovery and in signaling behaviors. Our new analysis reveals conflicting pressures on this ratio in symmetric and best-of-N decisions, which could be solved through a time-dependent signaling strategy. Additionally, our analysis suggests how ecological factors determining the density of suitable nest sites may have led to selective pressures for an optimal stable signaling ratio.

  16. Conservation hotspots for marine turtle nesting in the United States based on coastal development.

    PubMed

    Fuentes, Mariana M P B; Gredzens, Christian; Bateman, Brooke L; Boettcher, Ruth; Ceriani, Simona A; Godfrey, Matthew H; Helmers, David; Ingram, Dianne K; Kamrowski, Ruth L; Pate, Michelle; Pressey, Robert L; Radeloff, Volker C

    2016-12-01

    Coastal areas provide nesting habitat for marine turtles that is critical for the persistence of their populations. However, many coastal areas are highly affected by coastal development, which affects the reproductive success of marine turtles. Knowing the extent to which nesting areas are exposed to these threats is essential to guide management initiatives. This information is particularly important for coastal areas with both high nesting density and dense human development, a combination that is common in the United States. We assessed the extent to which nesting areas of the loggerhead (Caretta caretta), the green (Chelonia mydas), the Kemp's ridley (Lepidochelys kempii), and leatherback turtles (Dermochelys coriacea) in the continental United States are exposed to coastal development and identified conservation hotspots that currently have high reproductive importance and either face high exposure to coastal development (needing intervention), or have low exposure to coastal development, and are good candidates for continued and future protection. Night-time light, housing, and population density were used as proxies for coastal development and human disturbance. About 81.6% of nesting areas were exposed to housing and human population, and 97.8% were exposed to light pollution. Further, most (>65%) of the very high- and high-density nesting areas for each species/subpopulation, except for the Kemp's ridley, were exposed to coastal development. Forty-nine nesting sites were selected as conservation hotspots; of those high-density nesting sites, 49% were sites with no/low exposure to coastal development and the other 51% were exposed to high-density coastal development. Conservation strategies need to account for ~66.8% of all marine turtle nesting areas being on private land and for nesting sites being exposed to large numbers of seasonal residents. © 2016 by the Ecological Society of America.

  17. 3D deformable image matching: a hierarchical approach over nested subspaces

    NASA Astrophysics Data System (ADS)

    Musse, Olivier; Heitz, Fabrice; Armspach, Jean-Paul

    2000-06-01

    This paper presents a fast hierarchical method to perform dense deformable inter-subject matching of 3D MR Images of the brain. To recover the complex morphological variations in neuroanatomy, a hierarchy of 3D deformations fields is estimated, by minimizing a global energy function over a sequence of nested subspaces. The nested subspaces, generated from a single scaling function, consist of deformation fields constrained at different scales. The highly non linear energy function, describing the interactions between the target and the source images, is minimized using a coarse-to-fine continuation strategy over this hierarchy. The resulting deformable matching method shows low sensitivity to local minima and is able to track large non-linear deformations, with moderate computational load. The performances of the approach are assessed both on simulated 3D transformations and on a real data base of 3D brain MR Images from different individuals. The method has shown efficient in putting into correspondence the principle anatomical structures of the brain. An application to atlas-based MRI segmentation, by transporting a labeled segmentation map on patient data, is also presented.

  18. Effects of neonicotinoid imidacloprid exposure on bumble bee (Hymenoptera: Apidae) queen survival and nest initiation.

    PubMed

    Wu-Smart, Judy; Spivak, Marla

    2018-02-08

    Neonicotinoids are highly toxic to insects and may systemically translocate to nectar and pollen of plants where foraging bees may become exposed. Exposure to neonicotinoids can induce detrimental sublethal effects on individual and colonies of bees and may have long-term impacts, such as impaired foraging, reduced longevity, and reduced brood care or production. Less well-studied are the potential effects on queen bumble bees that may become exposed while foraging in the spring during colony initiation. This study assessed queen survival and nest founding in caged bumble bees [Bombus impatiens (Cresson) (Hymenoptera: Apidae)] after chronic (18-d) dietary exposure of imidacloprid in syrup (1, 5, 10, and 25 ppb) and pollen (0.3, 1.7, 3.3, and 8.3 ppb), paired respectively. Here we show some mortality in queens exposed at all doses even as low as 1 ppb, and, compared with untreated queens, significantly reduced survival of treated queens at the two highest doses. Queens that survived initial imidacloprid exposure commenced nest initiation; however, they exhibited dose-dependent delay in egg-laying and emergence of worker brood. Furthermore, imidacloprid treatment affected other parameters such as nest and queen weight. This study is the first to show direct impacts of imidacloprid at field-relevant levels on individual B. impatiens queen survival and nest founding, indicating that bumble bee queens are particularly sensitive to neonicotinoids when directly exposed. This study also helps focus pesticide risk mitigation efforts and highlights the importance of reducing exposure rates in the early spring when bumble bee queens, and other wild bees are foraging and initiating nests. © The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  19. Temperature-Dependent Sex Determination under Rapid Anthropogenic Environmental Change: Evolution at a Turtle's Pace?

    PubMed

    Refsnider, Jeanine M; Janzen, Fredric J

    2016-01-01

    Organisms become adapted to their environment by evolving through natural selection, a process that generally transpires over many generations. Currently, anthropogenically driven environmental changes are occurring orders of magnitude faster than they did prior to human influence, which could potentially outpace the ability of some organisms to adapt. Here, we focus on traits associated with temperature-dependent sex determination (TSD), a classic polyphenism, in a model turtle species to address the evolutionary potential of species with TSD to respond to rapid climate change. We show, first, that sex-ratio outcomes in species with TSD are sensitive to climatic variation. We then identify the evolutionary potential, in terms of heritability, of TSD and quantify the evolutionary potential of 3 key traits involved in TSD: pivotal temperature, maternal nest-site choice, and nesting phenology. We find that these traits display different patterns of adaptive potential: pivotal temperature exhibits moderate heritable variation, whereas nest-site choice and nesting phenology, with considerable phenotypic plasticity, have only modest evolutionary potential to alter sex ratios. Therefore, the most likely response of species with TSD to anthropogenically induced climate change may be a combination of microevolution in thermal sensitivity of the sex-determining pathway and of plasticity in maternal nesting behavior. © The American Genetic Association. 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Development of Nested PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Cylindrocladium scoparium on Eucalyptus.

    PubMed

    Qiao, Tian-Min; Zhang, Jing; Li, Shu-Jiang; Han, Shan; Zhu, Tian-Hui

    2016-10-01

    Eucalyptus dieback disease, caused by Cylindrocladium scoparium , has occurred in last few years in large Eucalyptus planting areas in China and other countries. Rapid, simple, and reliable diagnostic techniques are desired for the early detection of Eucalyptus dieback of C. scoparium prior to formulation of efficient control plan. For this purpose, three PCR-based methods of nested PCR, multiplex PCR, loop-mediated isothermal amplification (LAMP) were developed for detection of C. scoparium based on factor 1-alpha (tef1) and beta-tubulin gene in this study. All of the three methods showed highly specific to C. scoparium . The sensitivities of the nested PCR and LAMP were much higher than the multiplex PCR. The sensitivity of multiplex PCR was also higher than regular PCR. C. scoparium could be detected within 60 min from infected Eucalyptus plants by LAMP, while at least 2 h was needed by the rest two methods. Using different Eucalyptus tissues as samples for C. scoparium detection, all of the three PCR-based methods showed much better detection results than regular PCR. Base on the results from this study, we concluded that any of the three PCR-based methods could be used as diagnostic technology for the development of efficient strategies of Eucalyptus dieback disease control. Particularly, LAMP was the most practical method in field application because of its one-step and rapid reaction, simple operation, single-tube utilization, and simple visualization of amplification products.

  1. Development of Nested PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Cylindrocladium scoparium on Eucalyptus

    PubMed Central

    Qiao, Tian-Min; Zhang, Jing; Li, Shu-Jiang; Han, Shan; Zhu, Tian-Hui

    2016-01-01

    Eucalyptus dieback disease, caused by Cylindrocladium scoparium, has occurred in last few years in large Eucalyptus planting areas in China and other countries. Rapid, simple, and reliable diagnostic techniques are desired for the early detection of Eucalyptus dieback of C. scoparium prior to formulation of efficient control plan. For this purpose, three PCR-based methods of nested PCR, multiplex PCR, loop-mediated isothermal amplification (LAMP) were developed for detection of C. scoparium based on factor 1-alpha (tef1) and beta-tubulin gene in this study. All of the three methods showed highly specific to C. scoparium. The sensitivities of the nested PCR and LAMP were much higher than the multiplex PCR. The sensitivity of multiplex PCR was also higher than regular PCR. C. scoparium could be detected within 60 min from infected Eucalyptus plants by LAMP, while at least 2 h was needed by the rest two methods. Using different Eucalyptus tissues as samples for C. scoparium detection, all of the three PCR-based methods showed much better detection results than regular PCR. Base on the results from this study, we concluded that any of the three PCR-based methods could be used as diagnostic technology for the development of efficient strategies of Eucalyptus dieback disease control. Particularly, LAMP was the most practical method in field application because of its one-step and rapid reaction, simple operation, single-tube utilization, and simple visualization of amplification products. PMID:27721691

  2. Sensitive and Specific Target Sequences Selected from Retrotransposons of Schistosoma japonicum for the Diagnosis of Schistosomiasis

    PubMed Central

    Xu, Jing; Zhu, Xing-Quan; Wang, Sheng-Yue; Xia, Chao-Ming

    2012-01-01

    Background Schistosomiasis japonica is a serious debilitating and sometimes fatal disease. Accurate diagnostic tests play a key role in patient management and control of the disease. However, currently available diagnostic methods are not ideal, and the detection of the parasite DNA in blood samples has turned out to be one of the most promising tools for the diagnosis of schistosomiasis. In our previous investigations, a 230-bp sequence from the highly repetitive retrotransposon SjR2 was identified and it showed high sensitivity and specificity for detecting Schistosoma japonicum DNA in the sera of rabbit model and patients. Recently, 29 retrotransposons were found in S. japonicum genome by our group. The present study highlighted the key factors for selecting a new perspective sensitive target DNA sequence for the diagnosis of schistosomiasis, which can serve as example for other parasitic pathogens. Methodology/Principal Findings In this study, we demonstrated that the key factors based on the bioinformatic analysis for selecting target sequence are the higher genome proportion, repetitive complete copies and partial copies, and active ESTs than the others in the chromosome genome. New primers based on 25 novel retrotransposons and SjR2 were designed and their sensitivity and specificity for detecting S. japonicum DNA were compared. The results showed that a new 303-bp sequence from non-long terminal repeat (LTR) retrotransposon (SjCHGCS19) had high sensitivity and specificity. The 303-bp target sequence was amplified from the sera of rabbit model at 3 d post-infection by nested-PCR and it became negative at 17 weeks post-treatment. Furthermore, the percentage sensitivity of the nested-PCR was 97.67% in 43 serum samples of S. japonicum-infected patients. Conclusions/Significance Our findings highlighted the key factors based on the bioinformatic analysis for selecting target sequence from S. japonicum genome, which provide basis for establishing powerful molecular diagnostic techniques that can be used for monitoring early infection and therapy efficacy to support schistosomiasis control programs. PMID:22479661

  3. Application of multiplex nested methylated specific PCR in early diagnosis of epithelial ovarian cancer.

    PubMed

    Wang, Bi; Yu, Lei; Yang, Guo-Zhen; Luo, Xin; Huang, Lin

    2015-01-01

    To explore the application of multiplex nested methylated specific polymerase chain reaction (PCR) in the early diagnosis of epithelial ovarian carcinoma (EOC). Serum and fresh tissue samples were collected from 114 EOC patients. RUNX3, TFPI2 and OPCML served as target genes. Methylation levels of tissues were assessed by multiplex nested methylated specific PCR, the results being compared with those for carcinoma antigen 125 (CA125). The serum free deoxyribose nucleic acid (DNA) methylation spectrum of EOC patients was completely contained in the DNA spectrum of cancer tissues, providing an accurate reflection of tumor DNA methylation conditions. Serum levels of CA125 and free DNA methylation in the EOC group were evidently higher than those in benign lesion and control groups (p<0.05). Patients with early EOC had markedly lower serum CA125 than those with advanced EOC (p<0.05), but there was no significant difference in free DNA methylation (p>0.05). The sensitivity, specificity and positive predicative value (PPV) of multiplex nested methylated specific PCR were significantly higher for detection of all patients and those with early EOC than those for CA125 (p<0.05). In the detection of patients with advanced EOC, the PPV of CA125 detection was obviously lower than that of multiplex nested methylated specific PCR (p>0.05), but there was no significant difference in sensitivity (p>0.05). Serum free DNA methylation can be used as a biological marker for EOC and multiplex nested methylated specific PCR should be considered for early diagnosis since it can accurately determine tumor methylation conditions.

  4. Detection of Leishmania DNA in saliva among patients with HIV/AIDS in Trang Province, southern Thailand.

    PubMed

    Pandey, Netranapha; Siripattanapipong, Suradej; Leelayoova, Saovanee; Manomat, Jipada; Mungthin, Mathirut; Tan-Ariya, Peerapan; Bualert, Lertwut; Naaglor, Tawee; Siriyasatien, Padet; Phumee, Atchara; Piyaraj, Phunlerd

    2018-06-08

    Leishmaniasis is a neglected tropical disease causing opportunistic infection among patients with HIV/AIDS. The fatal form of this disease is visceral leishmaniasis (VL). DNA of Leishmania can be detected in saliva, for which the collection is noninvasive and requires little expertise. This study aimed to evaluate the sensitivity and specificity of a nested-PCR to amplify the Internal Transcribed Spacer 1 (ITS1) to detect Leishmania DNA in paired saliva and buffy coat samples of 305 Thai patients with HIV/AIDS in Trang Hospital, Trang Province, southern Thailand. For asymptomatic Leishmania infection among Thai patients with HIV/AIDS, the sensitivity and specificity of the nested-PCR-ITS1 in buffy coat were 73.9 and 100%, respectively. However, the sensitivity in saliva was 26.1% and specificity was 100%. Using the nested-PCR-ITS1, saliva and buffy coat samples showed positive agreement in only 52.0% of patients. Saliva tested results with the nested-PCR-ITS1 showed positive agreement with the Direct Agglutination Test (DAT) in 46.5% of patients. Only 12.1% of the samples showed positive agreement for Leishmania infection among all the three tests: saliva, buffy coat and DAT results. Using nucleotide sequencing, at least three species of Leishmania infection were identified in saliva, i.e., L. siamensis (n = 28), L. martiniquensis (n = 9), and L. donovani complex (n = 1). As a result, buffy coat still appears to be a better specimen to diagnose asymptomatic VL infection among individuals with HIV. However, the use of both buffy coat and saliva together as clinical specimens would increase the sensitivity of Leishmania detection. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Development and Validation of an Improved PCR Method Using the 23S-5S Intergenic Spacer for Detection of Rickettsiae in Dermacentor variabilis Ticks and Tissue Samples from Humans and Laboratory Animals

    PubMed Central

    Kakumanu, Madhavi L.; Ponnusamy, Loganathan; Sutton, Haley T.; Meshnick, Steven R.; Nicholson, William L.

    2016-01-01

    A novel nested PCR assay was developed to detect Rickettsia spp. in ticks and tissue samples from humans and laboratory animals. Primers were designed for the nested run to amplify a variable region of the 23S-5S intergenic spacer (IGS) of Rickettsia spp. The newly designed primers were evaluated using genomic DNA from 11 Rickettsia species belonging to the spotted fever, typhus, and ancestral groups and, in parallel, compared to other Rickettsia-specific PCR targets (ompA, gltA, and the 17-kDa protein gene). The new 23S-5S IGS nested PCR assay amplified all 11 Rickettsia spp., but the assays employing other PCR targets did not. The novel nested assay was sensitive enough to detect one copy of a cloned 23S-5S IGS fragment from “Candidatus Rickettsia amblyommii.” Subsequently, the detection efficiency of the 23S-5S IGS nested assay was compared to those of the other three assays using genomic DNA extracted from 40 adult Dermacentor variabilis ticks. The nested 23S-5S IGS assay detected Rickettsia DNA in 45% of the ticks, while the amplification rates of the other three assays ranged between 5 and 20%. The novel PCR assay was validated using clinical samples from humans and laboratory animals that were known to be infected with pathogenic species of Rickettsia. The nested 23S-5S IGS PCR assay was coupled with reverse line blot hybridization with species-specific probes for high-throughput detection and simultaneous identification of the species of Rickettsia in the ticks. “Candidatus Rickettsia amblyommii,” R. montanensis, R. felis, and R. bellii were frequently identified species, along with some potentially novel Rickettsia strains that were closely related to R. bellii and R. conorii. PMID:26818674

  6. Development and Validation of an Improved PCR Method Using the 23S-5S Intergenic Spacer for Detection of Rickettsiae in Dermacentor variabilis Ticks and Tissue Samples from Humans and Laboratory Animals.

    PubMed

    Kakumanu, Madhavi L; Ponnusamy, Loganathan; Sutton, Haley T; Meshnick, Steven R; Nicholson, William L; Apperson, Charles S

    2016-04-01

    A novel nested PCR assay was developed to detectRickettsiaspp. in ticks and tissue samples from humans and laboratory animals. Primers were designed for the nested run to amplify a variable region of the 23S-5S intergenic spacer (IGS) ofRickettsiaspp. The newly designed primers were evaluated using genomic DNA from 11Rickettsiaspecies belonging to the spotted fever, typhus, and ancestral groups and, in parallel, compared to otherRickettsia-specific PCR targets (ompA,gltA, and the 17-kDa protein gene). The new 23S-5S IGS nested PCR assay amplified all 11Rickettsiaspp., but the assays employing other PCR targets did not. The novel nested assay was sensitive enough to detect one copy of a cloned 23S-5S IGS fragment from "CandidatusRickettsia amblyommii." Subsequently, the detection efficiency of the 23S-5S IGS nested assay was compared to those of the other three assays using genomic DNA extracted from 40 adultDermacentor variabilisticks. The nested 23S-5S IGS assay detectedRickettsiaDNA in 45% of the ticks, while the amplification rates of the other three assays ranged between 5 and 20%. The novel PCR assay was validated using clinical samples from humans and laboratory animals that were known to be infected with pathogenic species ofRickettsia The nested 23S-5S IGS PCR assay was coupled with reverse line blot hybridization with species-specific probes for high-throughput detection and simultaneous identification of the species ofRickettsiain the ticks. "CandidatusRickettsia amblyommii,"R. montanensis,R. felis, andR. belliiwere frequently identified species, along with some potentially novelRickettsiastrains that were closely related toR. belliiandR. conorii. Copyright © 2016 Kakumanu et al.

  7. Distorting Gene Pools by Conservation: Assessing the Case of Doomed Turtle Eggs

    NASA Astrophysics Data System (ADS)

    Mrosovsky, N.

    2006-10-01

    Sea turtles have a high reproductive output and high mortality at early stages of the life cycle. In particular, many nests are laid below or close to high tide lines, and subsequently large numbers of eggs may be inundated and destroyed. A common conservation procedure is to relocate such doomed eggs to higher ground. This article examines this practice in the light of recent data revealing that some individual turtles tend to nest relatively near the water and others relatively higher up the beach. Discussion is focused on the question of why apparently poor placement of nests has not been selected against. Comparison between the ecology of leatherback and hawksbill turtle nesting beaches suggests that predictability of environmental conditions on the nesting beaches has an important influence on patterns of nest-site selection. Options are outlined for the management of nesting beaches where a high proportion of turtle eggs is subject to destruction by flooding.

  8. Distorting gene pools by conservation: Assessing the case of doomed turtle eggs.

    PubMed

    Mrosovsky, N

    2006-10-01

    Sea turtles have a high reproductive output and high mortality at early stages of the life cycle. In particular, many nests are laid below or close to high tide lines, and subsequently large numbers of eggs may be inundated and destroyed. A common conservation procedure is to relocate such doomed eggs to higher ground. This article examines this practice in the light of recent data revealing that some individual turtles tend to nest relatively near the water and others relatively higher up the beach. Discussion is focused on the question of why apparently poor placement of nests has not been selected against. Comparison between the ecology of leatherback and hawksbill turtle nesting beaches suggests that predictability of environmental conditions on the nesting beaches has an important influence on patterns of nest-site selection. Options are outlined for the management of nesting beaches where a high proportion of turtle eggs is subject to destruction by flooding.

  9. A Nested PCR Assay to Avoid False Positive Detection of the Microsporidian Enterocytozoon hepatopenaei (EHP) in Environmental Samples in Shrimp Farms

    PubMed Central

    Jaroenlak, Pattana; Sanguanrut, Piyachat; Williams, Bryony A. P.; Stentiford, Grant D.; Flegel, Timothy W.; Sritunyalucksana, Kallaya

    2016-01-01

    Hepatopancreatic microsporidiosis (HPM) caused by Enterocytozoon hepatopenaei (EHP) is an important disease of cultivated shrimp. Heavy infections may lead to retarded growth and unprofitable harvests. Existing PCR detection methods target the EHP small subunit ribosomal RNA (SSU rRNA) gene (SSU-PCR). However, we discovered that they can give false positive test results due to cross reactivity of the SSU-PCR primers with DNA from closely related microsporidia that infect other aquatic organisms. This is problematic for investigating and monitoring EHP infection pathways. To overcome this problem, a sensitive and specific nested PCR method was developed for detection of the spore wall protein (SWP) gene of EHP (SWP-PCR). The new SWP-PCR method did not produce false positive results from closely related microsporidia. The first PCR step of the SWP-PCR method was 100 times (104 plasmid copies per reaction vial) more sensitive than that of the existing SSU-PCR method (106 copies) but sensitivity was equal for both in the nested step (10 copies). Since the hepatopancreas of cultivated shrimp is not currently known to be infected with microsporidia other than EHP, the SSU-PCR methods are still valid for analyzing hepatopancreatic samples despite the lower sensitivity than the SWP-PCR method. However, due to its greater specificity and sensitivity, we recommend that the SWP-PCR method be used to screen for EHP in feces, feed and environmental samples for potential EHP carriers. PMID:27832178

  10. A Nested PCR Assay to Avoid False Positive Detection of the Microsporidian Enterocytozoon hepatopenaei (EHP) in Environmental Samples in Shrimp Farms.

    PubMed

    Jaroenlak, Pattana; Sanguanrut, Piyachat; Williams, Bryony A P; Stentiford, Grant D; Flegel, Timothy W; Sritunyalucksana, Kallaya; Itsathitphaisarn, Ornchuma

    2016-01-01

    Hepatopancreatic microsporidiosis (HPM) caused by Enterocytozoon hepatopenaei (EHP) is an important disease of cultivated shrimp. Heavy infections may lead to retarded growth and unprofitable harvests. Existing PCR detection methods target the EHP small subunit ribosomal RNA (SSU rRNA) gene (SSU-PCR). However, we discovered that they can give false positive test results due to cross reactivity of the SSU-PCR primers with DNA from closely related microsporidia that infect other aquatic organisms. This is problematic for investigating and monitoring EHP infection pathways. To overcome this problem, a sensitive and specific nested PCR method was developed for detection of the spore wall protein (SWP) gene of EHP (SWP-PCR). The new SWP-PCR method did not produce false positive results from closely related microsporidia. The first PCR step of the SWP-PCR method was 100 times (104 plasmid copies per reaction vial) more sensitive than that of the existing SSU-PCR method (106 copies) but sensitivity was equal for both in the nested step (10 copies). Since the hepatopancreas of cultivated shrimp is not currently known to be infected with microsporidia other than EHP, the SSU-PCR methods are still valid for analyzing hepatopancreatic samples despite the lower sensitivity than the SWP-PCR method. However, due to its greater specificity and sensitivity, we recommend that the SWP-PCR method be used to screen for EHP in feces, feed and environmental samples for potential EHP carriers.

  11. Evaluation of sensitivity of TaqMan RT-PCR for rubella virus detection in clinical specimens.

    PubMed

    Okamoto, Kiyoko; Mori, Yoshio; Komagome, Rika; Nagano, Hideki; Miyoshi, Masahiro; Okano, Motohiko; Aoki, Yoko; Ogura, Atsushi; Hotta, Chiemi; Ogawa, Tomoko; Saikusa, Miwako; Kodama, Hiroe; Yasui, Yoshihiro; Minagawa, Hiroko; Kurata, Takako; Kanbayashi, Daiki; Kase, Tetsuo; Murata, Sachiko; Shirabe, Komei; Hamasaki, Mitsuhiro; Kato, Takashi; Otsuki, Noriyuki; Sakata, Masafumi; Komase, Katsuhiro; Takeda, Makoto

    2016-07-01

    An easy and reliable assay for detection of the rubella virus is required to strengthen rubella surveillance. Although a TaqMan RT-PCR assay for detection of the rubella virus has been established in Japan, its utility for diagnostic purposes has not been tested. To allow introduction of the TaqMan RT-PCR into the rubella surveillance system in Japan, the sensitivity of the assay was determined using representative strains for all genotypes and clinical specimens. The detection limits of the method for individual genotypes were examined using viral RNA extracted from 13 representative strains. The assay was also tested at 10 prefectural laboratories in Japan, designated as local reference laboratories for measles and rubella, to allow nationwide application of the assay. The detection limits and amplification efficiencies of the assay were similar among all the representative strains of the 13 genotypes. The TaqMan RT-PCR could detect approximately 90% of throat swab and urine samples taken up to 5days of illness. These samples were determined positive by a highly sensitive nested RT-PCR. The TaqMan RT-PCR could detect at least 10 pfu of rubella virus. Although the sensitivity was somewhat lower than that of the conventional nested RT-PCR, the TaqMan RT-PCR could be more practical to routine tests for rubella laboratory diagnosis and detection in view of the rapid response and reducing risks of contamination. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Ground-nesting by the chimpanzees of the Nimba Mountains, Guinea: environmentally or socially determined?

    PubMed

    Koops, Kathelijne; Humle, Tatyana; Sterck, Elisabeth H M; Matsuzawa, Tetsuro

    2007-04-01

    The chimpanzees (Pan troglodytes verus) of the Nimba Mountains, Guinea, West Africa, commonly make both elaborate ("night") and simple ("day") nests on the ground. In this study we investigated which factors might influence ground-nesting in this population, and tested two ecological hypotheses: 1) climatic conditions, such as high wind speeds at high altitudes, may deter chimpanzees from nesting in trees; and 2) a lack of appropriate arboreal nesting opportunities may drive the chimpanzees to nest on the ground. In addition to testing these two hypotheses, we explored whether ground-nesting is a sex-linked behavior. Data were collected monthly between August 2003 and May 2004 along transects and ad libitum. To identify the sex of ground-nesting individuals, we used DNA extracted from hair samples. The results showed that the occurrence and distribution of ground nests were not affected by climatic conditions or a lack of appropriate nest trees. Support was found for the notion that ground-nesting is a sex-linked behavior, as males were responsible for building all of the elaborate ground nests and most of the simple ground nests sampled. Elaborate ground nests occurred mostly in nest groups associated with tree nests, whereas simple ground nests usually occurred without tree nests in their vicinity. These results suggest that ground-nesting may be socially, rather than ecologically, determined.

  13. Nest success of snowy plovers (Charadrius nivosus) in the Southern high plains of Texas

    USGS Publications Warehouse

    Saalfeld, S.T.; Conway, Warren C.; Haukos, D.A.; Johnson, W.P.

    2011-01-01

    Snowy Plovers (Charadrius nivosus) nesting on edges of saline lakes within the Southern High Plains (SHP) of Texas are threatened by habitat degradation due to reduced artesian spring flow, making many saline lakes unsuitable for nesting and migrating shorebirds. Factors influencing nest success were evaluated, current nest success estimates in the SHP of Texas were compared to estimates obtained ten years prior, and causes and timing of nest failures determined. Overall, 215 nests were monitored from three saline lakes in 20082009, with nest success estimates from Program MARK ranging from 7-33% ( x??= 22%). The leading causes of nest failures were attributed to predation (40%) and weather (36%). Nest success was negatively influenced by number of plants within 707-cm 2 plot, positively influenced by percent surface water availability, and at one saline lake, negatively influenced by day during the nesting season (i.e., nest success declined later in the nesting season). When compared to estimates ten years prior (19981999), mean nest success has declined by 31%. If nesting Snowy Plovers continue to experience increased predation rates, decreased hydrological integrity, and habitat alterations, populations will continue to decline throughout this region.

  14. Nest placement of the giant Amazon river turtle, Podocnemis expansa, in the Araguaia River, Goiás State, Brazil.

    PubMed

    Ferreira, Paulo Dias Júnior; Castro, Paulo de Tarso Amorim

    2005-05-01

    The giant Amazon river turtle (Podocnemis expansa) nests on extensive sand bars on the margins and interior of the channel during the dry season. The high concentration of nests in specific points of certain beaches indicates that the selection of nest placement is not random but is related to some geological aspects, such as bar margin inclination and presence of a high, sandy platform. The presence of access channels to high platform points or ramp morphology are decisive factors in the choice of nesting areas. The eroded and escarped margins of the beaches hinder the Amazon river turtle arriving at the most suitable places for nesting. Through the years, changes in beach morphology can alter nest distribution.

  15. Density-dependent nest predation in waterfowl: the relative importance of nest density versus nest dispersion

    USGS Publications Warehouse

    Ackerman, Joshua T.; Ringelman, Kevin M.; Eadie, J.M.

    2012-01-01

    When nest predation levels are very high or very low, the absolute range of observable nest success is constrained (a floor/ceiling effect), and it may be more difficult to detect density-dependent nest predation. Density-dependent nest predation may be more detectable in years with moderate predation rates, simply because there can be a greater absolute difference in nest success between sites. To test this, we replicated a predation experiment 10 years after the original study, using both natural and artificial nests, comparing a year when overall rates of nest predation were high (2000) to a year with moderate nest predation (2010). We found no evidence for density-dependent predation on artificial nests in either year, indicating that nest predation is not density-dependent at the spatial scale of our experimental replicates (1-ha patches). Using nearest-neighbor distances as a measure of nest dispersion, we also found little evidence for “dispersion-dependent” predation on artificial nests. However, when we tested for dispersion-dependent predation using natural nests, we found that nest survival increased with shorter nearest-neighbor distances, and that neighboring nests were more likely to share the same nest fate than non-adjacent nests. Thus, at small spatial scales, density-dependence appears to operate in the opposite direction as predicted: closer nearest neighbors are more likely to be successful. We suggest that local nest dispersion, rather than larger-scale measures of nest density per se, may play a more important role in density-dependent nest predation.

  16. Applying geographic profiling used in the field of criminology for predicting the nest locations of bumble bees.

    PubMed

    Suzuki-Ohno, Yukari; Inoue, Maki N; Ohno, Kazunori

    2010-07-21

    We tested whether geographic profiling (GP) can predict multiple nest locations of bumble bees. GP was originally developed in the field of criminology for predicting the area where an offender most likely resides on the basis of the actual crime sites and the predefined probability of crime interaction. The predefined probability of crime interaction in the GP model depends on the distance of a site from an offender's residence. We applied GP for predicting nest locations, assuming that foraging and nest sites were the crime sites and the offenders' residences, respectively. We identified the foraging and nest sites of the invasive species Bombus terrestris in 2004, 2005, and 2006. We fitted GP model coefficients to the field data of the foraging and nest sites, and used GP with the fitting coefficients. GP succeeded in predicting about 10-30% of actual nests. Sensitivity analysis showed that the predictability of the GP model mainly depended on the coefficient value of buffer zone, the distance at the mode of the foraging probability. GP will be able to predict the nest locations of bumble bees in other area by using the fitting coefficient values measured in this study. It will be possible to further improve the predictability of the GP model by considering food site preference and nest density. (c) 2010 Elsevier Ltd. All rights reserved.

  17. Clinical and epidemiological use of nested PCR targeting the repetitive element IS1111 associated with the transposase gene from Coxiella burnetii.

    PubMed

    Mares-Guia, Maria Angélica M M; Guterres, Alexandro; Rozental, Tatiana; Ferreira, Michelle Dos Santos; Lemos, Elba R S

    Q fever is a worldwide zoonosis caused by Coxiella burnetii-a small obligate intracellular Gram-negative bacterium found in a variety of animals. It is transmitted to humans by inhalation of contaminated aerosols from urine, feces, milk, amniotic fluid, placenta, abortion products, wool, and rarely by ingestion of raw milk from infected animals. Nested PCR can improve the sensitivity and specificity of testing while offering a suitable amplicon size for sequencing. Serial dilutions were performed tenfold to test the limit of detection, and the result was 10× detection of C. burnetti DNA with internal nested PCR primers relative to trans-PCR. Different biological samples were tested and identified only in nested PCR. This demonstrates the efficiency and effectiveness of the primers. Of the 19 samples, which amplify the partial sequence of C. burnetii, 12 were positive by conventional PCR and nested PCR. Seven samples-five spleen tissue samples from rodents and two tick samples-were only positive in nested PCR. With these new internal primers for trans-PCR, we demonstrate that our nested PCR assay for C. burnetii can achieve better results than conventional PCR. Published by Elsevier Editora Ltda.

  18. Selection of anthropogenic features and vegetation characteristics by nesting Common Ravens in the sagebrush ecosystem

    USGS Publications Warehouse

    Howe, Kristy B.; Coates, Peter S.; Delehanty, David J.

    2014-01-01

    Common Raven (Corvus corax) numbers and distribution are increasing throughout the sagebrush steppe, influencing avian communities in complex ways. Anthropogenic structures are thought to increase raven populations by providing food and nesting subsidies, which is cause for concern because ravens are important nest predators of sensitive species, including Greater Sage-Grouse (Centrocercus urophasianus). During 2007–2009, we located raven nests in southeastern Idaho and conducted a resource selection analysis. We measured variables at multiple spatial scales for 72 unique nest locations, including landscape-level vegetation characteristics and anthropogenic structures. Using generalized linear mixed models and an information-theoretic approach, we found a 31% decrease in the odds of nesting by ravens for every 1 km increase in distance away from a transmission line. Furthermore, a 100-m increase in distance away from the edge of two different land cover types decreased the odds of nesting by 20%, and an increase in the amount of edge by 1 km within an area of 102.1 ha centered on the nest increased the odds of nesting by 49%. A post hoc analysis revealed that ravens were most likely to nest near edges of adjoining big sagebrush (Artemisia tridentata) and land cover types that were associated with direct human disturbance or fire. These findings contribute to our understanding of raven expansion into rural environments and could be used to make better-informed conservation decisions, especially in the face of increasing renewable energy development.

  19. Rapid and sensitive multiplex single-tube nested PCR for the identification of five human Plasmodium species.

    PubMed

    Saito, Takahiro; Kikuchi, Aoi; Kaneko, Akira; Isozumi, Rie; Teramoto, Isao; Kimura, Masatsugu; Hirasawa, Noriyasu; Hiratsuka, Masahiro

    2018-06-01

    Malaria is caused by five species of Plasmodium in humans. Microscopy is currently used for pathogen detection, requiring considerable training and technical expertise as the parasites are often difficult to differentiate morphologically. Rapid diagnostic tests are as reliable as microscopy and offer faster diagnoses but possess lower detection limits and are incapable of distinguishing among the parasitic species. To improve global health efforts towards malaria control, a rapid, sensitive, species-specific, and economically viable diagnostic method is needed. In this study, we designed a malaria diagnostic method involving a multiplex single-tube nested PCR targeting Plasmodium mitochondrial cytochrome c oxidase III and single-stranded tag hybridization chromatographic printed-array strip. The detection sensitivity was found to be at least 40 times higher than that of agarose gel electrophoresis with ethidium bromide. This system also enables the identification of both single- and mixed-species malaria infections. The assay was validated with 152 Kenyan samples; using nested PCR as the standard, the assay's sensitivity and specificity were 88.7% and 100.0%, respectively. The turnaround time required, from PCR preparation to signal detection, is 90min. Our method should improve the diagnostic speed, treatment efficacy, and control of malaria, in addition to facilitating surveillance within global malaria eradication programs. Copyright © 2018 Elsevier B.V. All rights reserved.

  20. Arctic foxes, lemmings, and canada goose nest survival at cape Churchill, Manitoba

    USGS Publications Warehouse

    Reiter, M.E.; Andersen, D.E.

    2011-01-01

    We examined factors influencing Canada Goose (Branta canadensis interior) annual nest success, including the relative abundance of collared lemmings (Dicrostonyx richardsoni), arctic fox (Alopex lagopus) den occupancy, nest density, and spring phenology using data collected during annual Canada Goose breeding area surveys at Cape Churchill, Manitoba. Nest density and arctic fox den occupancy strongly influenced Canada Goose nest success. High nest density resulted in higher nest success and high den occupancy reduced nest success. Nest success was not influenced by lemming abundance in the current or previous year as predicted by the "bird-lemming" hypothesis. Reducing arctic fox abundance through targeted management increased nest survival of Canada Geese; a result that further emphasizes the importance of arctic fox as nest predators in this system. The spatial distribution of nest predators, at least for dispersed-nesting geese, may be most important for nest survival, regardless of the abundance of small mammals in the local ecosystem. Further understanding of the factors influencing the magnitude and variance in arctic fox abundance in this region, and the spatial scale at which these factors are realized, is necessary to fully explain predator-prey-alternative prey dynamics in this system. ?? 2011 by the Wilson Ornithological Society.

  1. Variation in Nest Temperatures of the American Alligator Found on the Kennedy Space Center Merritt Island National Wildlife Refuge

    NASA Technical Reports Server (NTRS)

    Lowers, Russell; Guillette, Louis J.; Weiss, Stephanie

    2016-01-01

    Information on nest temperatures of the American Alligator (Alligator mississippiensis) constructed in the wild is limited. Nesting temperatures during a critical thermal sensitive period determine the sex of alligators and are therefore critical in establishing the sex biases in recruitment efforts of alligators within a given community. Nest components, varying environmental conditions, and global warming could have a significant impact on nest temperatures, thus affecting future generations of a given population. One hundred and seventy four programmable thermistors were inserted into fifty eight nests from 2010 through 2015 nesting cycles. Three thermistors were placed inside each nest cavity (one on top of the eggs, one in the middle of the eggs, and one at the bottom of the clutch of the eggs) to collect temperature profiles in the incubation chamber and throughout the entire incubation period. One thermistor was also placed near or above these nests to obtain an ambient air temperature profile. Once retrieved, data from these thermistors were downloaded to examine temperature profiles throughout the incubation period as well as during the period of sexual determination. These data would help establish survival rates related to nest temperature and predict sex ratio of recruited neonates at the Kennedy Space Center. Over three million temperatures have been recorded since 2010 for the alligator thermistor study giving us insight to the recruitment efforts found here. Precipitation was the largest influence on nesting temperatures outside of daily photoperiod, with immediate changes of up to eight degrees Celsius.

  2. Wetland and microhabitat use by nesting four-toed salamanders in Maine

    USGS Publications Warehouse

    Chalmers, R.J.; Loftin, C.S.

    2006-01-01

    Little is known of Four-Toed Salamander (Hemidactylium scutatum) habitat use, despite the species' extensive range and elevated conservation status. We investigated species-habitat relationships that predict H. scutatum nesting presence in Maine at wetland and microhabitat scales by comparing microhabitats with and without nests. We created logistic regression models, selected models with AIC, and evaluated models with reserve data. Wetlands with nests were best predicted by shoreline microhabitat of Sphagnum spp., wood substrate, water flow, blue-joint reed grass (Calamagrostis canadensis), meadowsweet (Spiraea alba), steeplebush (Spiraea tomentosa), sensitive fern (Onoclea sensibilis), and absence of sheep laurel (Kalmia angustifolia) or deciduous forest canopy. Within occupied wetlands, shoreline microhabitat where nests occurred was best distinguished from available, unoccupied shoreline microhabitat by steeper shore, greater near-shore and basin water depth, deeper nesting vegetation, presence of moss spp. and winterberry (Ilex verticillata), and a negative association with S. alba, leatherleaf (Chamaedaphne calyculata), and K. angustifolia. These models of wetland and microhabitat use by H. scutatum may assist ecologists and managers in detecting and conserving this species. Copyright 2006 Society for the Study of Amphibians and Reptiles.

  3. Specific and sensitive detection of the conifer pathogen Gremmeniella abietina by nested PCR

    PubMed Central

    Zeng, Qing-Yin; Hansson, Per; Wang, Xiao-Ru

    2005-01-01

    Background Gremmeniella abietina (Lagerb.) Morelet is an ascomycete fungus that causes stem canker and shoot dieback in many conifer species. The fungus is widespread and causes severe damage to forest plantations in Europe, North America and Asia. To facilitate early diagnosis and improve measures to control the spread of the disease, rapid, specific and sensitive detection methods for G. abietina in conifer hosts are needed. Results We designed two pairs of specific primers for G. abietina based on the 18S rDNA sequence variation pattern. These primers were validated against a wide range of fungi and 14 potential conifer hosts. Based on these specific primers, two nested PCR systems were developed. The first system employed universal fungal primers to enrich the fungal DNA targets in the first round, followed by a second round selective amplification of the pathogen. The other system employed G. abietina-specific primers in both PCR steps. Both approaches can detect the presence of G. abietina in composite samples with high sensitivity, as little as 7.5 fg G. abietina DNA in the host genomic background. Conclusion The methods described here are rapid and can be applied directly to a wide range of conifer species, without the need for fungal isolation and cultivation. Therefore, it represents a promising alternative to disease inspection in forest nurseries, plantations and quarantine control facilities. PMID:16280082

  4. Varying hydric conditions during incubation influence egg water exchange and hatchling phenotype in the red-eared slider turtle.

    PubMed

    Delmas, Virginie; Bonnet, Xavier; Girondot, Marc; Prévot-Julliard, Anne-Caroline

    2008-01-01

    Environmental conditions within the nest, notably temperature and moisture of substrate, exert a powerful influence during embryogenesis in oviparous reptiles. The influence of fluctuating nest temperatures has been experimentally examined in different reptile species; however, similar experiments using moisture as the key variable are lacking. In this article, we examine the effect of various substrate moisture regimes during incubation on different traits (egg mass, incubation length, and hatchling mass) in a chelonian species with flexible-shelled eggs, the red-eared slider turtle (Trachemys scripta elegans). Our results show that the rate of water uptake by the eggs was higher in wet than in dry substrate and varied across development. More important, during the first third of development, the egg mass changes were relatively independent of the soil moisture level; they became very sensitive to moisture levels during the other two-thirds. Moreover, hydric conditions exerted a strong influence on the eggs' long-term sensitivity to the moisture of the substrate. Even short-term episodes of high or low levels of moisture modified permanently their water sensitivity, notably through modification of eggshell shape and volume, and in turn entailed significant effects on hatchling mass (and hence offspring quality). Such complex influences of fluctuating moisture levels at various incubation stages on hatchling phenotype better reflect the natural situation, compared to experiments based on stable, albeit different, moisture levels.

  5. Sharp-Tailed Grouse Nest Survival and Nest Predator Habitat Use in North Dakota's Bakken Oil Field.

    PubMed

    Burr, Paul C; Robinson, Aaron C; Larsen, Randy T; Newman, Robert A; Ellis-Felege, Susan N

    2017-01-01

    Recent advancements in extraction technologies have resulted in rapid increases of gas and oil development across the United States and specifically in western North Dakota. This expansion of energy development has unknown influences on local wildlife populations and the ecological interactions within and among species. Our objectives for this study were to evaluate nest success and nest predator dynamics of sharp-tailed grouse (Tympanuchus phasianellus) in two study sites that represented areas of high and low energy development intensities in North Dakota. During the summers of 2012 and 2013, we monitored 163 grouse nests using radio telemetry. Of these, 90 nests also were monitored using miniature cameras to accurately determine nest fates and identify nest predators. We simultaneously conducted predator surveys using camera scent stations and occupancy modeling to estimate nest predator occurrence at each site. American badgers (Taxidea taxus) and striped skunks (Mephitis mephitis) were the primary nest predators, accounting for 56.7% of all video recorded nest depredations. Nests in our high intensity gas and oil area were 1.95 times more likely to succeed compared to our minimal intensity area. Camera monitored nests were 2.03 times more likely to succeed than non-camera monitored nests. Occupancy of mammalian nest predators was 6.9 times more likely in our study area of minimal gas and oil intensity compared to the high intensity area. Although only a correlative study, our results suggest energy development may alter the predator community, thereby increasing nest success for sharp-tailed grouse in areas of intense development, while adjacent areas may have increased predator occurrence and reduced nest success. Our study illustrates the potential influences of energy development on the nest predator-prey dynamics of sharp-tailed grouse in western North Dakota and the complexity of evaluating such impacts on wildlife.

  6. Nesting ecology of boreal forest birds following a massive outbreak of spruce beetles

    USGS Publications Warehouse

    Matsuoka, S.M.; Handel, C.M.

    2007-01-01

    We studied breeding dark-eyed juncos (Junco hyemalis), yellow-rumped warblers (Dendroica coronata), and spruce-nesting birds from 1997 to 1998 among forests with different levels of spruce (Picea spp.) mortality following an outbreak of spruce beetles (Dendroctonus rufipennis) in Alaska, USA. We identified species using live and beetle-killed spruce for nest sites and monitored nests to determine how the outbreak influenced avian habitat selection and reproduction. We tested predictions that 1) nesting success of ground-nesting juncos would increase with spruce mortality due to proliferation of understory vegetation available to conceal nests from predators, 2) nesting success of canopy-nesting warblers would decrease with spruce mortality due to fewer live spruce in which to conceal nests, and 3) both species would alter nest-site selection in response to disturbance. Juncos did not benefit from changes in understory vegetation; nesting success in highly disturbed stands (46%) was comparable to that in undisturbed habitats throughout their range. In stands with low spruce mortality, nesting success of juncos was low (5%) and corresponded with high densities of red squirrels (Tamiasciurus hudsonicus). Yellow-rumped warblers nested exclusively in spruce, but success did not vary with spruce mortality. As disturbance increased, nesting warblers switched from selecting forest patches with high densities of live white spruce (Picea glauca) to patches with beetle-killed spruce. Warblers also placed nests in large-diameter live or beetle-killed spruce, depending on which was more abundant in the stand, with no differences in nesting success. Five of the 12 other species of spruce-nesting birds also used beetle-killed spruce as nest sites. Because beetle-killed spruce can remain standing for >50 years, even highly disturbed stands provide an important breeding resource for boreal forest birds. We recommend that boreal forest managers preserve uncut blocks of infested forest within managed forest landscapes and practice partial harvest of beetle-killed spruce rather than commercial clear-cutting of infested stands in order to sustain breeding bird populations until natural reforestation occurs. Because breeding densities do not always reflect fitness, assessing impacts of a massive natural disturbance should include measuring impacts of changes in vegetation on both reproductive success and predator–prey dynamics.

  7. Design of a Nested Eight-Channel Sodium and Four-Channel Proton Coil for 7 Tesla Knee Imaging

    PubMed Central

    Brown, Ryan; Madelin, Guillaume; Lattanzi, Riccardo; Chang, Gregory; Regatte, Ravinder R.; Sodickson, Daniel K.; Wiggins, Graham C.

    2012-01-01

    The critical design aim for a dual-tuned sodium/proton coil is to maximize sodium sensitivity and transmit field (B1+) homogeneity while simultaneously providing adequate proton sensitivity and homogeneity. While most dual-frequency coils utilize lossy high-impedance trap circuits or PIN diodes to allow dual-resonance, we explored a nested-coil design for sodium/proton knee imaging at 7T. A stand-alone eight-channel sodium receive array was implemented without standard dual-resonance circuitry to provide improved sodium signal-to-noise ratio (SNR) over a volume coil. A detunable sodium birdcage was added for homogeneous sodium excitation and a four-channel proton transmit-receive array was added to provide anatomical reference imaging and B0 shimming capability. Both modules were implemented with minimal disturbance to the eight-channel sodium array by managing their respective resonances and geometrical arrangement. In vivo sodium SNR was 1.2 to 1.7 times greater in the developed eight-channel array than in a mono-nuclear sodium birdcage coil, while the developed four-channel proton array provided SNR similar to that of a commercial mono-nuclear proton birdcage coil. PMID:22887123

  8. Urine-Based Nested PCR for the Diagnosis of Mycobacterium tuberculosis: A Comparative Study Between HIV-Positive and HIV-Negative Patients.

    PubMed

    Jamshidi Makiani, Mahin; Davoodian, Parivash; Baghershiroodi, Mahnaz; Nejatizadeh, Abdol Azim; Fakkhar, Farideh; Zangeneh, Mehrangiz; Jahangiri, Nadia

    2016-08-01

    While tuberculosis (TB) can be diagnosed by microscopy and culture, the sensitivity of Ziehl-Neelsen staining is variable and culture results require 4 - 8 weeks to be determined. Polymerase chain reaction (PCR) and its modifications, including nested PCR, might be promising methods for the rapid diagnosis of TB. This study aimed to evaluate the performance of nested PCR on urine samples of human immunodeficiency virus (HIV)-positive and -negative patients with different manifestations of clinical TB. In a prospective study, three early-morning urine samples from 100 patients with pulmonary TB (PTB) or extrapulmonary TB (EPTB) were evaluated using a molecular target with insertion element IS6110, specific to the Mycobacterium tuberculosis genome, and nested PCR was performed. The results were analyzed with SPSS version 22. A total of 100 patients, including 74 (74%) with PTB and 26 (26%) with EPTB, were enrolled. Positive smears were seen in 38 patients (38%). Lymph nodes were the most commonly involved organ in 14 of the 26 (53.8%) EPTB patients (13.5%). Seven (23.1%) of the EPTB patients were HIV-positive. Urine PCR was positive in only 28 patients (28%). Seven HIV-positive patients with PTB showed positive urine PCR results. Moreover, PCR results were positive in only one of the seven HIV-positive subjects with EPTB. Positive PCR results were found in 20 of the 73 HIV-negative patients (27.4%) and in 8 of the 27 HIV-positive patients (29.6%). Therefore, there was no significant difference between the HIV-negative and HIV-positive patients for urine PCR (sensitivity 29.6%, specificity 72.6%; positive and negative predictive values 28% and 72%, respectively; P = 0.138). Nested PCR showed the same sensitivity in HIV-positive and HIV-negative patients. It can be applied as a rapid technique for the diagnosis of TB.

  9. Comparison of real-time SYBR green dengue assay with real-time taqman RT-PCR dengue assay and the conventional nested PCR for diagnosis of primary and secondary dengue infection

    PubMed Central

    Paudel, Damodar; Jarman, Richard; Limkittikul, Kriengsak; Klungthong, Chonticha; Chamnanchanunt, Supat; Nisalak, Ananda; Gibbons, Robert; Chokejindachai, Watcharee

    2011-01-01

    Background: Dengue fever and dengue hemorrhagic fever are caused by dengue virus. Dengue infection remains a burning problem of many countries. To diagnose acute dengue in the early phase we improve the low cost, rapid SYBR green real time assay and compared the sensitivity and specificity with real time Taqman® assay and conventional nested PCR assay. Aims: To develop low cost, rapid and reliable real time SYBR green diagnostic dengue assay and compare with Taqman real-time assay and conventional nested PCR (modified Lanciotti). Materials and Methods: Eight cultured virus strains were diluted in tenth dilution down to undetectable level by the PCR to optimize the primer, temperature (annealing, and extension and to detect the limit of detection of the assay. Hundred and ninety three ELISA and PCR proved dengue clinical samples were tested with real time SYBR® Green assay, real time Taqman® assay to compare the sensitivity and specificity. Results: Sensitivity and specificity of real time SYBR® green dengue assay (84% and 66%, respectively) was almost comparable to those (81% and 74%) of Taqman real time PCR dengue assay. Real time SYBR® green RT-PCR was equally sensitive in primary and secondary infection while real time Taqman was less sensitive in the secondary infection. Sensitivity of real time Taqman on DENV3 (87%) was equal to SYBR green real time PCR dengue assay. Conclusion: We developed low cost rapid diagnostic SYBR green dengue assay. Further study is needed to make duplex primer assay for the serotyping of dengue virus. PMID:22363089

  10. Selection of nest-site habitat by interior least terns in relation to sandbar construction

    USGS Publications Warehouse

    Sherfy, M.H.; Stucker, J.H.; Buhl, D.A.

    2012-01-01

    Federally endangered interior least terns (Sternula antillarum) nest on bare or sparsely vegetated sandbars on midcontinent river systems. Loss of nesting habitat has been implicated as a cause of population declines, and managing these habitats is a major initiative in population recovery. One such initiative involves construction of mid-channel sandbars on the Missouri River, where natural sandbar habitat has declined in quantity and quality since the late 1990s. We evaluated nest-site habitat selection by least terns on constructed and natural sandbars by comparing vegetation, substrate, and debris variables at nest sites (na =a 798) and random points (na =a 1,113) in bare or sparsely vegetated habitats. Our logistic regression models revealed that a broader suite of habitat features was important in nest-site selection on constructed than on natural sandbars. Odds ratios for habitat variables indicated that avoidance of habitat features was the dominant nest-site selection process on both sandbar types, with nesting terns being attracted to nest-site habitat features (gravel and debris) and avoiding vegetation only on constructed sandbars, and avoiding silt and leaf litter on both sandbar types. Despite the seemingly uniform nature of these habitats, our results suggest that a complex suite of habitat features influences nest-site choice by least terns. However, nest-site selection in this social, colonially nesting species may be influenced by other factors, including spatial arrangement of bare sand habitat, proximity to other least terns, and prior habitat occupancy by piping plovers (Charadrius melodus). We found that nest-site selection was sensitive to subtle variation in habitat features, suggesting that rigor in maintaining habitat condition will be necessary in managing sandbars for the benefit of least terns. Further, management strategies that reduce habitat features that are avoided by least terns may be the most beneficial to nesting least terns. ?? 2011 The Wildlife Society.

  11. Factors influencing nest survival and productivity of Red-throated Loons (Gavia stellata) in Alaska

    USGS Publications Warehouse

    Rizzolo, Daniel; Schmutz, Joel A.; McCloskey, Sarah E.; Fondell, Thomas F.

    2014-01-01

    Red-throated Loon (Gavia stellata) numbers in Alaska have fluctuated dramatically over the past 3 decades; however, the demographic processes contributing to these population dynamics are poorly understood. To examine spatial and temporal variation in productivity, we estimated breeding parameters at 5 sites in Alaska: at Cape Espenberg and the Copper River Delta we estimated nest survival, and at 3 sites within the Yukon-Kuskokwim Delta we estimated nest survival and productivity. Nest survival varied broadly among sites and years; annual estimates (lower, upper 95% confidence interval) ranged from 0.09 (0.03, 0.29) at Cape Espenberg in 2001 to 0.93 (0.76, 0.99) at the Copper River Delta in 2002. Annual variation among sites was not concordant, suggesting that site-scale factors had a strong influence on nest survival. Models of nest survival indicated that visits to monitor nests had a negative effect on nest daily survival probability, which if not accounted for biased nest survival strongly downward. The sensitivity of breeding Red-throated Loons to nest monitoring suggests other sources of disturbance that cause incubating birds to flush from their nests may also reduce nest survival. Nest daily survival probability at the Yukon-Kuskokwim Delta was negatively associated with an annual index of fox occurrence. Survival through the incubation and chick-rearing periods on the Yukon-Kuskokwim Delta ranged from 0.09 (0.001, 0.493) to 0.50 (0.04, 0.77). Daily survival probability during the chick-rearing period was lower for chicks that had a sibling in 2 of 3 years, consistent with the hypothesis that food availability was limited. Estimates of annual productivity on the Yukon-Kuskokwim Delta ranged from 0.17 to 1.0 chicks per pair. Productivity was not sufficient to maintain population stability in 2 of 3 years, indicating that nest depredation by foxes and poor foraging conditions during chick rearing can have important effects on productivity.

  12. Selection of nest-site habitat by interior least terns in relation to sandbar construction

    USGS Publications Warehouse

    Sherfy, Mark H.; Stucker, Jennifer H.; Buhl, Deborah A.

    2012-01-01

    Federally endangered interior least terns (Sternula antillarum) nest on bare or sparsely vegetated sandbars on midcontinent river systems. Loss of nesting habitat has been implicated as a cause of population declines, and managing these habitats is a major initiative in population recovery. One such initiative involves construction of mid-channel sandbars on the Missouri River, where natural sandbar habitat has declined in quantity and quality since the late 1990s. We evaluated nest-site habitat selection by least terns on constructed and natural sandbars by comparing vegetation, substrate, and debris variables at nest sites (n = 798) and random points (n = 1,113) in bare or sparsely vegetated habitats. Our logistic regression models revealed that a broader suite of habitat features was important in nest-site selection on constructed than on natural sandbars. Odds ratios for habitat variables indicated that avoidance of habitat features was the dominant nest-site selection process on both sandbar types, with nesting terns being attracted to nest-site habitat features (gravel and debris) and avoiding vegetation only on constructed sandbars, and avoiding silt and leaf litter on both sandbar types. Despite the seemingly uniform nature of these habitats, our results suggest that a complex suite of habitat features influences nest-site choice by least terns. However, nest-site selection in this social, colonially nesting species may be influenced by other factors, including spatial arrangement of bare sand habitat, proximity to other least terns, and prior habitat occupancy by piping plovers (Charadrius melodus). We found that nest-site selection was sensitive to subtle variation in habitat features, suggesting that rigor in maintaining habitat condition will be necessary in managing sandbars for the benefit of least terns. Further, management strategies that reduce habitat features that are avoided by least terns may be the most beneficial to nesting least terns.

  13. Nested polymerase chain reaction (PCR) targeting 16S rDNA for bacterial identification in empyema.

    PubMed

    Prasad, Rajniti; Kumari, Chhaya; Das, B K; Nath, Gopal

    2014-05-01

    Empyema in children causes significant morbidity and mortality. However, identification of organisms is a major concern. To detect bacterial pathogens in pus specimens of children with empyema by 16S rDNA nested polymerase chain reaction (PCR) and correlate it with culture and sensitivity. Sixty-six children admitted to the paediatric ward with a diagnosis of empyema were enrolled prospectively. Aspirated pus was subjected to cytochemical examination, culture and sensitivity, and nested PCR targeting 16S rDNA using a universal eubacterial primer. Mean (SD) age was 5·8 (1·8) years (range 1-13). Analysis of aspirated pus demonstrated total leucocyte count >1000×10(6)/L, elevated protein (≧20 g/L) and decreased glucose (≤2·2 mmol/L) in 80·3%, 98·5% and 100%, respectively. Gram-positive cocci were detected in 29 (43·9%) and Gram-negative bacilli in two patients. Nested PCR for the presence of bacterial pathogens was positive in 50·0%, compared with 36·3% for culture. 16S rDNA PCR improves rates of detection of bacteria in pleural fluid, and can detect bacterial species in a single assay as well as identifying unusual and unexpected causal agents.

  14. Site selection and nest survival of the Bar-Headed Goose (Anser indicus) on the Mongolian Plateau

    USGS Publications Warehouse

    Batbayar, Nyambayar; Takekawa, John Y.; Natsagdorj, Tseveenmyadag; Spragens, Kyle A.; Xiao, Xiamgming

    2014-01-01

    Waterbirds breeding on the Mongolian Plateau in Central Asia must find suitable wetland areas for nesting in a semiarid region characterized by highly variable water conditions. The first systematic nesting study of a waterbird dependent on this region for breeding was conducted on the Bar-headed Goose (Anser indicus). The purpose of this study was to document Bar-headed Goose nesting locations, characterize nests and nesting strategies, and estimate daily nest survival (n = 235 nests) from eight areas of west-central Mongolia across three summers (2009–2011) using a modified Mayfield estimator. Bar-headed Goose daily nest survival ranged from 0.94 to 0.98, with a 3-year average nest success of 42.6% during incubation. Bar-headed Geese were found to primarily nest on isolated pond and lake islands as previously reported, but were also documented regularly, though less frequently, along rocky cliffs in several regions of west-central Mongolia. Daily nest survival was higher for cliff nests than for island nests. Information-theoretic models indicated that nest survival decreased with nest age and varied annually with changing environmental conditions. Results of this study suggest that while Bar-headed Geese primarily rely on nesting island sites these sites may be more susceptible to anthropogenic disturbance and predation events influenced by seasonal variation in environmental conditions, and that higher daily nest survival values documented for the less frequent cliff nest strategy may provide an important alternative strategy during poor island nest success years. Thus, conservation efforts for this and other waterbird species in the semiarid region should be focused on conserving nesting islands and protecting them from disturbance in areas of high livestock densities experiencing a rapidly warming climate.

  15. Nest sanitation elicits egg discrimination in cuckoo hosts.

    PubMed

    Yang, Canchao; Chen, Min; Wang, Longwu; Liang, Wei; Møller, Anders Pape

    2015-11-01

    Nest sanitation is a nearly universal behavior in birds, while egg discrimination is a more specific adaptation that has evolved to counter brood parasitism. These two behaviors are closely related with nest sanitation being the ancestral behavior, and it has been hypothesized to constitute a preadaptation for egg discrimination. However, previous studies found little evidence to support this hypothesis. Here, we conducted an empirical test of the association between nest sanitation and egg discrimination in the barn swallow (Hirundo rustica) by inserting a single non-mimetic model egg or a non-mimetic model egg plus half a peanut shell into host nests. Compared to the rejection rate of single model eggs, barn swallows significantly increased egg rejection frequency if a half peanut shell was simultaneously introduced. Our result for the first time shows the impact of nest sanitation on egg discrimination and demonstrates that nest sanitation can elicit egg discrimination in hosts of brood parasites. This study provided evidence for nest sanitation being a preadaptation to egg discrimination by facilitating egg rejection, thereby significantly advancing our understanding of avian cognition of foreign objects. Furthermore, we suggest that egg discrimination behavior in many accepters and intermediate rejecters may be lost or diluted. Such egg discrimination can be elicited and restored after nest sanitation, implying a sensitive and rapid phenotypic response to increased risk of parasitism. Our study offers a novel perspective for investigating the role of so-called intermediate rejecter individuals or species in the long-term coevolutionary cycle between brood parasites and their hosts.

  16. A Newly Developed Nested PCR Assay for the Detection of Helicobacter pylori in the Oral Cavity.

    PubMed

    Ismail, Hawazen; Morgan, Claire; Griffiths, Paul; Williams, John; Jenkins, Gareth

    2016-01-01

    To develop a new nested polymerase chain reaction (PCR) assay for identifying Helicobacter pylori DNA from dental plaque. H. pylori is one of the most common chronic bacterial pathogens in humans. The accurate detection of this organism is essential for proper patient management and for the eradication of the bacteria following treatment. Forty-nine patients (24 males and 25 females; mean age: 51; range, 19 to 94 y) were investigated for the presence of H. pylori in dental plaque by single-step PCR and nested PCR and in the stomach by single-step PCR, nested PCR, and histologic examination. The newly developed nested PCR assay identified H. pylori DNA in gastric biopsies of 18 patients who were histologically classified as H. pylori-positive and 2 additional biopsies of patients who were H. pylori-negative by histologic examination (20/49; 40.8%). Dental plaque samples collected before and after endoscopy from the 49 patients revealed that single-step PCR did not detect H. pylori but nested PCR was able to detect H. pylori DNA in 40.8% (20/49) patients. Nested PCR gave a higher detection rate (40.8%, 20/49) than that of histology (36.7%, 18/49) and single-step PCR. When nested PCR results were compared with histology results there was no significant difference between the 2 methods. Our newly developed nested PCR assay is at least as sensitive as histology and may be useful for H. pylori detection in patients unfit for endoscopic examination.

  17. High contrast sensitivity for visually guided flight control in bumblebees.

    PubMed

    Chakravarthi, Aravin; Kelber, Almut; Baird, Emily; Dacke, Marie

    2017-12-01

    Many insects rely on vision to find food, to return to their nest and to carefully control their flight between these two locations. The amount of information available to support these tasks is, in part, dictated by the spatial resolution and contrast sensitivity of their visual systems. Here, we investigate the absolute limits of these visual properties for visually guided position and speed control in Bombus terrestris. Our results indicate that the limit of spatial vision in the translational motion detection system of B. terrestris lies at 0.21 cycles deg -1 with a peak contrast sensitivity of at least 33. In the perspective of earlier findings, these results indicate that bumblebees have higher contrast sensitivity in the motion detection system underlying position control than in their object discrimination system. This suggests that bumblebees, and most likely also other insects, have different visual thresholds depending on the behavioral context.

  18. A productivity model for parasitized, multibrooded songbirds

    USGS Publications Warehouse

    Powell, L.A.; Knutson, M.G.

    2006-01-01

    We present an enhancement of a simulation model to predict annual productivity for Wood Thrushes (Hylocichla mustelina) and American Redstarts (Setophaga ruticilla); the model includes effects of Brown-headed Cowbird (Molothrus ater) parasitism. We used species-specific data from the Driftless Area Ecoregion of Wisconsin, Minnesota, and Iowa to parameterize the model as a case study. The simulation model predicted annual productivity of 2.03 ?? 1.60 SD for Wood Thrushes and 1.56 ?? 1.31 SD for American Redstarts. Our sensitivity analysis showed that high parasitism lowered Wood Thrush annual productivity more than American Redstart productivity, even though parasitism affected individual nests of redstarts more severely. Annual productivity predictions are valuable for habitat managers, but productivity is not easily obtained from field studies. Our model provides a useful means of integrating complex life history parameters to predict productivity for songbirds that experience nest parasitism. ?? The Cooper Ornithological Society 2006.

  19. Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals.

    PubMed

    Zhang, Xin; He, Chuan-Chuan; Liu, Jin-Ming; Li, Hao; Lu, Ke; Fu, Zhi-Qiang; Zhu, Chuan-Gang; Liu, Yi-Ping; Tong, Lai-Bao; Zhou, De-Bao; Zha, Li; Hong, Yang; Jin, Ya-Mei; Lin, Jiao-Jiao

    2017-04-13

    Schistosomiasis japonica is a common zoonosis. Domestic animals are the primary source of infection and play an important role in disease transmission. The prevalence and infectivity of this disease in domestic animals in China have significantly decreased and, for this reason, diagnostics with a higher sensitivity have become increasingly necessary. It was reported that polymerase chain reaction (PCR)-based methods could be used to detect schistosome infection in humans and animals and presented a high sensitivity and specificity. The present study aimed to develop a PCR-based method for detection of Schistosoma japonicum infection in domestic animals. A specific nested-PCR assay was developed to detect S. japonicum infection in domestic animals via amplification of a 231-bp DNA fragment of retrotransposon SjR2. The developed assay was first used in sera and dry blood filter paper (DBFP) from goats and buffaloes at different time points of infection. Then, 78 DBFPs from 39 artificially-infected bovines at 14 and 28 days post-infection and 42 DBFPs from schistosome-negative bovines from the city of Huangshan in the Anhui province were used to evaluate the diagnostic validity. Furthermore, this assay was used to detect S. japonicum infection in domestic animals in Dongzhi and Wangjiang counties. The expected PCR product was detected in eggs and adult worms of S. japonicum and blood samples from S. japonicum-infected goats and water buffaloes, but not from Fasciola and Haemonchus contortus worms. The nested-PCR assay could detect the target S. japonicum DNA in DBFPs from goats and buffaloes after day 3 post-infection. The sensitivity in buffaloes at 14 and 28 days post-infection was 92.30% (36/39) and 100% (39/39), respectively. The specificity was 97.60% (41/42). The positivity rates in Dongzhi and Wangjiang counties were 6.00% and 8.00% in bovines and 22.00% and 16.67% in goats, respectively. The positivity rates in goats in both counties were higher than those in bovines with a significant difference in Dongzhi County but not in Wangjiang County (P < 0.05 and P = 0.23, respectively). Our results suggest that the developed nested-PCR assay may be used for the diagnosis of S. japonicum infection in domestic animals, and the control of S. japonicum infection in goats should be paid more attention.

  20. Reproductive ecology and demography of the 'Akohekohe

    USGS Publications Warehouse

    Simon, John C.; Pratt, Thane K.; Berlin, Kim E.; Kowalsky, James R.

    2001-01-01

    The ‘Ākohekohe (Palmeria dolei) is an endangered Hawaiian honeycreeper endemic to the montane rain forests of east Maui in the Hawaiian Islands. We investigated ‘Ākohekohe nesting ecology using color-banded birds for the first time as a background to understanding the species' conservation. From 1994–1997, we color-banded 78 individuals, located and monitored 46 active nests, and took behavioral data during 534 hr of nest observation at Hanawī Natural Area Reserve, near the center of the species' range. ‘Ākohekohe nesting behavior and life history closely resembled that of ‘Apapane (Himatione sanguinea) and related honeycreepers. The birds were monogamous within and among years, and we found no evidence of polyandry, polygyny, or helpers at the nest. The nesting season extended from November to early June. Females performed all incubation and brooding. Males provisioned females and nestlings, and they were more active than females in feeding fledglings during the two-week period of parental dependency. Modal clutch size, as determined from egg counts at three nests and by counting begging chicks at other nests, was two eggs, and parents frequently fledged two chicks. We found an overall nest success rate of 68% by the Mayfield method, a high rate compared with other Hawaiian honeycreepers and continental passerines. An average of 1.1 chicks fledged per active nest, and at least 42% of nesting pairs made two or more nesting attempts per season. Rats (Rattus spp.) were abundant at the study site, and we confirmed their depredating some ‘Ākohekohe nests, so we did not expect to find such a high rate of nest success. The estimated annual probability of adult survival was also high, at 0.95 ± 0.10 (SE).

  1. Nested polymerase chain reaction on blood clots for gene encoding 56 kDa antigen and serology for the diagnosis of scrub typhus.

    PubMed

    Prakash, J A J; Kavitha, M L; Mathai, E

    2011-01-01

    Scrub typhus is a zoonotic illness endemic in the Asia-Pacific region. Early diagnosis and appropriate management contribute significantly to preventing adverse outcomes including mortality. Serology is widely used for diagnosing scrub typhus. Recent reports suggest that polymerase chain reaction (PCR) could be a rapid and reliable alternative. This study assessed the utility of these tests for scrub typhus diagnosis. Nested PCR to detect the 56 kDa antigen gene of O. tsutsugamushi was performed on blood clots from 87 individuals with clinically suspected scrub typhus. Weil-Felix test and scrub typhus IgM ELISA were performed on serum samples from the same patients. As a gold standard reference test was not available, latent class analysis (LCA) was used to assess the performance of the three tests. The LCA analysis showed the sensitivity of Weil-Felix test, IgM ELISA and PCR to be 59%, 100% and 58% respectively. The specificity of ELISA was only 73%, whereas those of the Weil-Felix test and PCR were 94% and 100% respectively. Nested PCR using blood clots while specific, lacked sensitivity as compared to IgM ELISA. In resource-poor settings Weil-Felix test still remains valuable despite its moderate sensitivity.

  2. Daily Activity and Nest Occupation Patterns of Fox Squirrels (Sciurus niger) throughout the Year.

    PubMed

    Wassmer, Thomas; Refinetti, Roberto

    2016-01-01

    The authors investigated the general activity and nest occupation patterns of fox squirrels in a natural setting using temperature-sensitive data loggers that measure activity as changes in the microenvironment of the animal. Data were obtained from 25 distinct preparations, upon 14 unique squirrels, totaling 1385 recording days. The animals were clearly diurnal, with a predominantly unimodal activity pattern, although individual squirrels occasionally exhibited bimodal patterns, particularly in the spring and summer. Even during the short days of winter (9 hours of light), the squirrels typically left the nest after dawn and returned before dusk, spending only about 7 hours out of the nest each day. Although the duration of the daily active phase did not change with the seasons, the squirrels exited the nest earlier in the day when the days became longer in the summer and exited the nest later in the day when the days became shorter in the winter, thus tracking dawn along the seasons. During the few hours spent outside the nest each day, fox squirrels seemed to spend most of the time sitting or lying. These findings suggest that fox squirrels may have adopted a slow life history strategy that involves long periods of rest on trees and short periods of ground activity each day.

  3. High energy electrons beyond 100 GEV observed by emulsion chamber

    NASA Technical Reports Server (NTRS)

    Nishimura, J.; Fujii, M.; Yoshida, A.; Taira, T.; Aizu, H.; Nomura, Y.; Kobayashi, T.; Kazuno, M.; Nishio, A.; Golden, R. L.

    1986-01-01

    Much efforts have been expended to observe the spectrum of electrons in the high energy region with large area emulsion chambers exposed at balloon altitudes, and now 15 electrons beyond 1 TeV have been observed. The observed integral flux at 1 TeV is (3.24 + or - 0.87)x10(-5)/sq m sec sr. The statistics of the data around a few hundred GeV are also improving by using new shower detecting films of high sensitivity. The astrophysical significance of the observed spectrum are discussed for the propagation of electrons based on the leaky box and the nested leaky box model.

  4. In situ detection of the Clostridium botulinum type C1 toxin gene in wetland sediments with a nested PCR assay

    USGS Publications Warehouse

    Williamson, Judy L.; Rocke, Tonie E.; Aiken, Judd M.

    1999-01-01

    A nested PCR was developed for detection of the Clostridium botulinum type C1 toxin gene in sediments collected from wetlands where avian botulism outbreaks had or had not occurred. The C1 toxin gene was detected in 16 of 18 sites, demonstrating both the ubiquitous distribution of C. botulinum type C in wetland sediments and the sensitivity of the detection assay.

  5. Assessing nest-building behavior of mice using a 3D depth camera.

    PubMed

    Okayama, Tsuyoshi; Goto, Tatsuhiko; Toyoda, Atsushi

    2015-08-15

    We developed a novel method to evaluate the nest-building behavior of mice using an inexpensive depth camera. The depth camera clearly captured nest-building behavior. Using three-dimensional information from the depth camera, we obtained objective features for assessing nest-building behavior, including "volume," "radius," and "mean height". The "volume" represents the change in volume of the nesting material, a pressed cotton square that a mouse shreds and untangles in order to build its nest. During the nest-building process, the total volume of cotton fragments is increased. The "radius" refers to the radius of the circle enclosing the fragments of cotton. It describes the extent of nesting material dispersion. The "radius" averaged approximately 60mm when a nest was built. The "mean height" represents the change in the mean height of objects. If the nest walls were high, the "mean height" was also high. These features provided us with useful information for assessment of nest-building behavior, similar to conventional methods for the assessment of nest building. However, using the novel method, we found that JF1 mice built nests with higher walls than B6 mice, and B6 mice built nests faster than JF1 mice. Thus, our novel method can evaluate the differences in nest-building behavior that cannot be detected or quantified by conventional methods. In future studies, we will evaluate nest-building behaviors of genetically modified, as well as several inbred, strains of mice, with several nesting materials. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Regional Model Nesting Within GFS Daily Forecasts Over West Africa

    NASA Technical Reports Server (NTRS)

    Druyan, Leonard M.; Fulakeza, Matthew; Lonergan, Patrick; Worrell, Ruben

    2010-01-01

    The study uses the RM3, the regional climate model at the Center for Climate Systems Research of Columbia University and the NASA/Goddard Institute for Space Studies (CCSR/GISS). The paper evaluates 30 48-hour RM3 weather forecasts over West Africa during September 2006 made on a 0.5 grid nested within 1 Global Forecast System (GFS) global forecasts. September 2006 was the Special Observing Period #3 of the African Monsoon Multidisciplinary Analysis (AMMA). Archived GFS initial conditions and lateral boundary conditions for the simulations from the US National Weather Service, National Oceanographic and Atmospheric Administration were interpolated four times daily. Results for precipitation forecasts are validated against Tropical Rainfall Measurement Mission (TRMM) satellite estimates and data from the Famine Early Warning System (FEWS), which includes rain gauge measurements, and forecasts of circulation are compared to reanalysis 2. Performance statistics for the precipitation forecasts include bias, root-mean-square errors and spatial correlation coefficients. The nested regional model forecasts are compared to GFS forecasts to gauge whether nesting provides additional realistic information. They are also compared to RM3 simulations driven by reanalysis 2, representing high potential skill forecasts, to gauge the sensitivity of results to lateral boundary conditions. Nested RM3/GFS forecasts generate excessive moisture advection toward West Africa, which in turn causes prodigious amounts of model precipitation. This problem is corrected by empirical adjustments in the preparation of lateral boundary conditions and initial conditions. The resulting modified simulations improve on the GFS precipitation forecasts, achieving time-space correlations with TRMM of 0.77 on the first day and 0.63 on the second day. One realtime RM3/GFS precipitation forecast made at and posted by the African Centre of Meteorological Application for Development (ACMAD) in Niamey, Niger is shown.

  7. [Brant goose colonies near snowy owls: internest distances in relation to lemming and arctic fox abundance].

    PubMed

    Kharitonov, S P; Volkov, A E; Willems, F; van Kleef, H; Klaassen, R H G; Nowak, D J; Nowak, A I; Bublichenko, A G

    2008-01-01

    Brant goose colonies around snowy owl nests have been studied near Meduza Bay (73 degrees 21' N, 80 degrees 32' E) and in the lower reaches of the Uboinaya River (73 degrees 37' N, 82 degrees 10' E), the northwestern Taimyr Peninsula, from 1999 to 2006. All brant nests within 680 m from an owl nest have been regarded as an individual colony. The results show that the area of the colony is always larger than the guarded area around the owl nest. In years of high abundance of lemmings, brant geese nest generally closer to the owl nest than in years of high abundance. When arctic foxes are abundant, however, brant geese nest significantly closer to owls than when the foxes are scarce, irrespective of lemming abundance. The mechanism of brant colony formation around owl nests is based on a number of stimuli.

  8. Decline in a population of spectacled eiders nesting on the Yukon-Kuskokwim Delta, Alaska

    USGS Publications Warehouse

    Ely, Craig R.; Dau, Christian; Babcock, Christopher

    1994-01-01

    The number of spectacled eiders nesting on two study areas near the Kashunuk River, on the central Yukon-Kuskokwim (Y-K) Delta, Alaska, declined by over 75% in the last 20 years. Nesting densities have remained low, but have not significantly declined since 1985. There has been no decrease in the reproductive effort of individual females as indicated by average clutch sizes. There has been a significant decline in the proportion of nests located on islands on one of the two study areas. Nesting success declined significantly during the 1970's. Success was not monitored in recent years, but has likely been low, based on the poor nesting success and declining numbers of cackling Canada geese and black brant nesting on the area. Nest predation by arctic foxes severely limited the productivity of cackling Canada geese, and foxes were likely the major predators of eider nests. Persistent high predation rates may lead to local extirpation in highly philopatric species such as eiders.

  9. Projected response of an endangered marine turtle population to climate change

    NASA Astrophysics Data System (ADS)

    Saba, Vincent S.; Stock, Charles A.; Spotila, James R.; Paladino, Frank V.; Tomillo, Pilar Santidrián

    2012-11-01

    Assessing the potential impacts of climate change on individual species and populations is essential for the stewardship of ecosystems and biodiversity. Critically endangered leatherback turtles in the eastern Pacific Ocean are excellent candidates for such an assessment because their sensitivity to contemporary climate variability has been substantially studied. If incidental fisheries mortality is eliminated, this population still faces the challenge of recovery in a rapidly changing climate. Here we combined an Earth system model, climate model projections assessed by the Intergovernmental Panel on Climate Change and a population dynamics model to estimate a 7% per decade decline in the Costa Rica nesting population over the twenty-first century. Whereas changes in ocean conditions had a small effect on the population, the ~2.5°C warming of the nesting beach was the primary driver of the decline through reduced hatching success and hatchling emergence rate. Hatchling sex ratio did not substantially change. Adjusting nesting phenology or changing nesting sites may not entirely prevent the decline, but could offset the decline rate. However, if future observations show a long-term decline in hatching success and emergence rate, anthropogenic climate mitigation of nests (for example, shading, irrigation) may be able to preserve the nesting population.

  10. Comparative evaluation of the nested ITS PCR against the 18S PCR-RFLP in a survey of bovine trypanosomiasis in Kwale County, Kenya.

    PubMed

    Odongo, Steven; Delespaux, Vincent; Ngotho, Maina; Bekkele, Serkalem Mindaye; Magez, Stefan

    2016-09-01

    We compared the nested internal transcribed spacer (ITS) PCR and the 18S PCR-RFLP (restriction-fragment length polymorphism) pan-trypanosome assays in a cross-sectional survey of bovine trypanosomiasis in 358 cattle in Kwale County, Kenya. The prevalence of trypanosomiasis as determined by the nested ITS PCR was 19.6% (70/358) and by 18S PCR-RFLP was 16.8% (60/358). Of the pathogenic trypanosomes detected, the prevalence of Trypanosoma congolense and Trypanosoma vivax was greater than that of Trypanosoma simiae The nested ITS PCR detected 83 parasite events, whereas the 18S PCR-RFLP detected 64; however, overall frequencies of infections and the parasite events detected did not differ between the assays (χ(2) = 0.8, df = 1, p > 0.05 and χ(2) = 2.5, df = 1, p > 0.05, respectively). The kappa statistic (0.8) showed good agreement between the tests. The nested ITS PCR and the 18S PCR-RFLP had comparable sensitivity, although the nested ITS PCR was better at detecting mixed infections (χ(2) = 5.4, df = 1, p < 0.05). © 2016 The Author(s).

  11. Olive Ridley Sea Turtle Hatching Success as a Function of the Microbial Abundance in Nest Sand at Ostional, Costa Rica

    PubMed Central

    Bézy, Vanessa S.; Valverde, Roldán A.; Plante, Craig J.

    2015-01-01

    Several studies have suggested that significant embryo mortality is caused by microbes, while high microbial loads are generated by the decomposition of eggs broken by later nesting turtles. This occurs commonly when nesting density is high, especially during mass nesting events (arribadas). However, no previous research has directly quantified microbial abundance and the associated effects on sea turtle hatching success at a nesting beach. The aim of this study was to test the hypothesis that the microbial abundance in olive ridley sea turtle nest sand affects the hatching success at Ostional, Costa Rica. We applied experimental treatments to alter the microbial abundance within the sand into which nests were relocated. We monitored temperature, oxygen, and organic matter content throughout the incubation period and quantified the microbial abundance within the nest sand using a quantitative polymerase chain reaction (qPCR) molecular analysis. The most successful treatment in increasing hatching success was the removal and replacement of nest sand. We found a negative correlation between hatching success and fungal abundance (fungal 18S rRNA gene copies g-1 nest sand). Of secondary importance in determining hatching success was the abundance of bacteria (bacterial 16S rRNA gene copies g-1 g-1 nest sand). Our data are consistent with the hypothesis that high microbial activity is responsible for the lower hatching success observed at Ostional beach. Furthermore, the underlying mechanism appears to be the deprivation of oxygen and exposure to higher temperatures resulting from microbial decomposition in the nest. PMID:25714355

  12. Degenerate and Nested PCR: a Highly Sensitive and Specific Method for Detection of Human Papillomavirus Infection in Cutaneous Warts

    PubMed Central

    Harwood, Catherine A.; Spink, Patricia J.; Surentheran, T.; Leigh, Irene M.; de Villiers, Ethel-Michele; McGregor, Jane M.; Proby, Charlotte M.; Breuer, Judith

    1999-01-01

    The role of human papillomavirus (HPV) in anogenital carcinogenesis is firmly established, but evidence that supports a similar role in skin remains speculative. Immunosuppressed renal transplant recipients have an increased incidence of viral warts and nonmelanoma skin cancer, and the presence of HPV DNA in these lesions, especially types associated with the condition epidermodysplasia verruciformis (EV), has led to suggestions that HPV may play a pathogenic role. However, differences in the specificities and sensitivities of techniques used to detect HPV in skin have led to wide discrepancies in the spectrum of HPV types reported. We describe a degenerate nested PCR technique with the capacity to detect a broad spectrum of cutaneous, mucosal, and EV HPV types. In a series of 51 warts from 23 renal transplant recipients, this method detected HPV DNA in all lesions, representing a significant improvement over many previously published studies. Cutaneous types were found in 84.3% of warts and EV types were found in 80.4% of warts, whereas mucosal types were detected in 27.4% of warts. In addition, the method allowed codetection of two or more distinct HPV types in 94.1% of lesions. In contrast, single HPV types were detected in all but 1 of 20 warts from 15 immunocompetent individuals. In summary, we have established a highly sensitive and comprehensive degenerate PCR methodology for detection and genotyping of HPV from the skin and have demonstrated a diverse spectrum of multiple HPV types in cutaneous warts from transplant recipients. Studies designed to assess the significance of these findings to cutaneous carcinogenesis are under way. PMID:10523550

  13. Detecting DNA methylation of the BCL2, CDKN2A and NID2 genes in urine using a nested methylation specific polymerase chain reaction assay to predict bladder cancer.

    PubMed

    Scher, Michael B; Elbaum, Michael B; Mogilevkin, Yakov; Hilbert, David W; Mydlo, Jack H; Sidi, A Ami; Adelson, Martin E; Mordechai, Eli; Trama, Jason P

    2012-12-01

    Detection of methylated DNA has been shown to be a good biomarker for bladder cancer. Bladder cancer has the highest recurrence rate of any cancer and, as such, patients are regularly monitored using invasive diagnostic techniques. As urine is easily attainable, bladder cancer is an optimal cancer to detect using DNA methylation. DNA methylation is highly specific in cancer detection. However, it is difficult to detect because of the limited amount of DNA present in the urine of patients with bladder cancer. Therefore, an improved, sensitive and noninvasive diagnostic test is needed. We developed a highly specific and sensitive nested methylation specific polymerase chain reaction assay to detect the presence of bladder cancer in small volumes of patient urine. The genes assayed for DNA methylation are BCL2, CDKN2A and NID2. The regions surrounding the DNA methylation sites were amplified in a methylation independent first round polymerase chain reaction and the amplification product from the first polymerase chain reaction was used in a real-time methylation specific polymerase chain reaction. Urine samples were collected from patients receiving treatment at Wolfson Medical Center in Holon, Israel. In a pilot clinical study using patient urine samples we were able to differentiate bladder cancer from other urogenital malignancies and nonmalignant conditions with a sensitivity of 80.9% and a specificity of 86.4%. We developed a novel methylation specific polymerase chain reaction assay for the detection and monitoring of bladder cancer using DNA extracted from patient urine. The assay may also be combined with other diagnostic tests to improve accuracy. Copyright © 2012 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  14. Anatomy of a bottleneck: diagnosing factors limiting population growth in the Puerto Rican parrot

    USGS Publications Warehouse

    Beissenger, S.R.; Wunderle, J.M.; Meyers, J.M.; Saether, B.-E.; Engen, S.

    2008-01-01

    The relative importance of genetic, demographic, environmental, and catastrophic processes that maintain population bottlenecks has received little consideration. We evaluate the role of these factors in maintaining the Puerto Rican Parrot (Amazona vittata) in a prolonged bottleneck from 1973 through 2000 despite intensive conservation efforts. We first conduct a risk analysis, then examine evidence for the importance of specific processes maintaining the bottleneck using the multiple competing hypotheses approach, and finally integrate these results through a sensitivity analysis of a demographic model using life-stage simulation analysis (LSA) to determine the relative importance of genetic, demographic, environmental, and catastrophic processes on population growth. Annual population growth has been slow and variable (1.0 6 5.2 parrots per year, or an average k?1.05 6 0.19) from 16 parrots (1973) to a high of 40-42 birds (1997-1998). A risk analysis based on population prediction intervals (PPI) indicates great risk and large uncertainty, with a range of 22?83 birds in the 90% PPI only five years into the future. Four primary factors (reduced hatching success due to inbreeding, failure of adults to nest, nest failure due to nongenetic causes, and reduced survival of adults and juveniles) were responsible for maintaining the bottleneck. Egghatchability rates were low (70.6% per egg and 76.8% per pair), and hatchability increased after mate changes, suggesting inbreeding effects. Only an average of 34% of the population nested annually, which was well below the percentage of adults that should have reached an age of first breeding (41-56%). This chronic failure to nest appears to have been caused primarily by environmental and/or behavioral factors, and not by nest-site scarcity or a skewed sex ratio. Nest failure rates from nongenetic causes (i.e., predation, parasitism, and wet cavities) were low (29%) due to active management (protecting nests and fostering captive young into wild nests), diminishing the importance of nest failure as a limiting factor. Annual survival has been periodically reduced by catastrophes (hurricanes), which have greatly constrained population growth, but survival rates were high under non-catastrophic conditions. Although the importance of factors maintaining the Puerto Rican Parrot bottleneck varied throughout the 30-year period of study, we determined their long-term influence using LSA simulations to correlate variation in demographic rates with variation in population growth (k). The bottleneck appears to have been maintained primarily by periodic catastrophes (hurricanes) that reduced adult survival, and secondarily by environmental and/or behavioral factors that resulted in a failure of many adults to nest. The influence of inbreeding through reduced hatching success played a much less significant role, even when additional effects of inbreeding on the production and mortality of young were incorporated into the LSA. Management actions needed to speed recovery include (1) continued nest guarding to minimize the effects of nest failure due to nongenetic causes; (2) creating a second population at another location on the island --a process that was recently initiated--to reduce the chance that hurricane strikes will cause extinction; and (3) determining the causes of the low percentage of breeders in the population and ameliorating them, which would have a large impact on population growth.

  15. Nest sanitation through defecation: antifungal properties of wood cockroach feces.

    PubMed

    Rosengaus, Rebeca B; Mead, Kerry; Du Comb, William S; Benson, Ryan W; Godoy, Veronica G

    2013-11-01

    The wood cockroach Cryptocercus punctulatus nests as family units inside decayed wood, a substrate known for its high microbial load. We tested the hypothesis that defecation within their nests, a common occurrence in this species, reduces the probability of fungal development. Conidia of the entomopathogenic fungus, Metarhizium anisopliae, were incubated with crushed feces and subsequently plated on potato dextrose agar. Relative to controls, the viability of fungal conidia was significantly reduced following incubation with feces and was negatively correlated with incubation time. Although the cockroach's hindgut contained abundant β-1,3-glucanase activity, its feces had no detectable enzymatic function. Hence, these enzymes are unlikely the source of the fungistasis. Instead, the antifungal compound(s) of the feces involved heat-sensitive factor(s) of potential microbial origin. When feces were boiled or when they were subjected to ultraviolet radiation and subsequently incubated with conidia, viability was "rescued" and germination rates were similar to those of controls. Filtration experiments indicate that the fungistatic activity of feces results from chemical interference. Because Cryptocercidae cockroaches have been considered appropriate models to make inferences about the factors fostering the evolution of termite sociality, we suggest that nesting in microbe-rich environments likely selected for the coupling of intranest defecation and feces fungistasis in the common ancestor of wood cockroaches and termites. This might in turn have served as a preadaptation that prevented mycosis as these phylogenetically related taxa diverged and evolved respectively into subsocial and eusocial organizations.

  16. Comparison of three human papillomavirus DNA detection methods: Next generation sequencing, multiplex-PCR and nested-PCR followed by Sanger based sequencing.

    PubMed

    da Fonseca, Allex Jardim; Galvão, Renata Silva; Miranda, Angelica Espinosa; Ferreira, Luiz Carlos de Lima; Chen, Zigui

    2016-05-01

    To compare the diagnostic performance for HPV infection using three laboratorial techniques. Ninty-five cervicovaginal samples were randomly selected; each was tested for HPV DNA and genotypes using 3 methods in parallel: Multiplex-PCR, the Nested PCR followed by Sanger sequencing, and the Next_Gen Sequencing (NGS) with two assays (NGS-A1, NGS-A2). The study was approved by the Brazilian National IRB (CONEP protocol 16,800). The prevalence of HPV by the NGS assays was higher than that using the Multiplex-PCR (64.2% vs. 45.2%, respectively; P = 0.001) and the Nested-PCR (64.2% vs. 49.5%, respectively; P = 0.003). NGS also showed better performance in detecting high-risk HPV (HR-HPV) and HPV16. There was a weak interobservers agreement between the results of Multiplex-PCR and Nested-PCR in relation to NGS for the diagnosis of HPV infection, and a moderate correlation for HR-HPV detection. Both NGS assays showed a strong correlation for detection of HPVs (k = 0.86), HR-HPVs (k = 0.91), HPV16 (k = 0.92) and HPV18 (k = 0.91). NGS is more sensitive than the traditional Sanger sequencing and the Multiplex PCR to genotype HPVs, with promising ability to detect multiple infections, and may have the potential to establish an alternative method for the diagnosis and genotyping of HPV. © 2015 Wiley Periodicals, Inc.

  17. Whole blood Nested PCR and Real-time PCR amplification of Talaromyces marneffei specific DNA for diagnosis.

    PubMed

    Lu, Sha; Li, Xiqing; Calderone, Richard; Zhang, Jing; Ma, Jianchi; Cai, Wenying; Xi, Liyan

    2016-02-01

    Talaromyces marneffei is a dimorphic pathogenic fungus, which is a life-threatening invasive mycosis in the immunocompromised host. Prompt diagnosis of T. marneffei infection remains difficult although there has been progress in attempts to expedite the diagnosis of this infection. We previously demonstrated the value of nested polymerase chain reaction (PCR) to detect T. marneffei in paraffin embedded tissue samples with high sensitivity and specificity. In this study, this assay was used to detect the DNA of T. marneffei in whole blood samples. Real-time PCR assay was also evaluated to identify T. marneffei in the same samples. Twenty out of 30 whole blood samples (67%) collected from 23 patients were found positive by using the nested PCR assay, while 23/30 (77%) samples were found positive by using the real-time PCR assay. In order to express accurately the fungal loads, we used a normalized linearized plasmid as an internal control for real-time PCR. The assay results were correlated as the initial quantity (copies/μl) with fungal burden. These data indicate that combination of nested PCR and real-time PCR assay provides an attractive alternative for identification of T. marneffei DNA in whole blood samples of HIV-infected patients. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Nest sanitation through defecation: antifungal properties of wood cockroach feces

    NASA Astrophysics Data System (ADS)

    Rosengaus, Rebeca B.; Mead, Kerry; Du Comb, William S.; Benson, Ryan W.; Godoy, Veronica G.

    2013-11-01

    The wood cockroach Cryptocercus punctulatus nests as family units inside decayed wood, a substrate known for its high microbial load. We tested the hypothesis that defecation within their nests, a common occurrence in this species, reduces the probability of fungal development. Conidia of the entomopathogenic fungus, Metarhizium anisopliae, were incubated with crushed feces and subsequently plated on potato dextrose agar. Relative to controls, the viability of fungal conidia was significantly reduced following incubation with feces and was negatively correlated with incubation time. Although the cockroach's hindgut contained abundant β-1,3-glucanase activity, its feces had no detectable enzymatic function. Hence, these enzymes are unlikely the source of the fungistasis. Instead, the antifungal compound(s) of the feces involved heat-sensitive factor(s) of potential microbial origin. When feces were boiled or when they were subjected to ultraviolet radiation and subsequently incubated with conidia, viability was "rescued" and germination rates were similar to those of controls. Filtration experiments indicate that the fungistatic activity of feces results from chemical interference. Because Cryptocercidae cockroaches have been considered appropriate models to make inferences about the factors fostering the evolution of termite sociality, we suggest that nesting in microbe-rich environments likely selected for the coupling of intranest defecation and feces fungistasis in the common ancestor of wood cockroaches and termites. This might in turn have served as a preadaptation that prevented mycosis as these phylogenetically related taxa diverged and evolved respectively into subsocial and eusocial organizations.

  19. Sites of ozone sensitivity in diverse maize inbred lines

    USDA-ARS?s Scientific Manuscript database

    Tropospheric ozone (O3) is an air pollutant that costs ~$14-26 billion in global crop losses and is projected to worsen in the future. Potential sites of O3 sensitivity in maize were tested by growing 200 inbred lines, including the nested association mapping population founder lines, under ambient...

  20. Nested high-resolution large-eddy simulations in WRF to support wind power

    NASA Astrophysics Data System (ADS)

    Mirocha, J.; Kirkil, G.; Kosovic, B.; Lundquist, J. K.

    2009-12-01

    The WRF model’s grid nesting capability provides a potentially powerful framework for simulating flow over a wide range of scales. One such application is computation of realistic inflow boundary conditions for large eddy simulations (LES) by nesting LES domains within mesoscale domains. While nesting has been widely and successfully applied at GCM to mesoscale resolutions, the WRF model’s nesting behavior at the high-resolution (Δx < 1000m) end of the spectrum is less well understood. Nesting LES within msoscale domains can significantly improve turbulent flow prediction at the scale of a wind park, providing a basis for superior site characterization, or for improved simulation of turbulent inflows encountered by turbines. We investigate WRF’s grid nesting capability at high mesh resolutions using nested mesoscale and large-eddy simulations. We examine the spatial scales required for flow structures to equilibrate to the finer mesh as flow enters a nest, and how the process depends on several parameters, including grid resolution, turbulence subfilter stress models, relaxation zones at nest interfaces, flow velocities, surface roughnesses, terrain complexity and atmospheric stability. Guidance on appropriate domain sizes and turbulence models for LES in light of these results is provided This work is performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344 LLNL-ABS-416482

  1. Detection of Histoplasma capsulatum from clinical specimens by cycling probe-based real-time PCR and nested real-time PCR.

    PubMed

    Muraosa, Yasunori; Toyotome, Takahito; Yahiro, Maki; Watanabe, Akira; Shikanai-Yasuda, Maria Aparecida; Kamei, Katsuhiko

    2016-05-01

    We developed new cycling probe-based real-time PCR and nested real-time PCR assays for the detection of Histoplasma capsulatum that were designed to detect the gene encoding N-acetylated α-linked acidic dipeptidase (NAALADase), which we previously identified as an H. capsulatum antigen reacting with sera from patients with histoplasmosis. Both assays specifically detected the DNAs of all H. capsulatum strains but not those of other fungi or human DNA. The limited of detection (LOD) of the real-time PCR assay was 10 DNA copies when using 10-fold serial dilutions of the standard plasmid DNA and 50 DNA copies when using human serum spiked with standard plasmid DNA. The nested real-time PCR improved the LOD to 5 DNA copies when using human serum spiked with standard plasmid DNA, which represents a 10-fold higher than that observed with the real-time PCR assay. To assess the ability of the two assays to diagnose histoplasmosis, we analyzed a small number of clinical specimens collected from five patients with histoplasmosis, such as sera (n = 4), formalin-fixed paraffin-embedded (FFPE) tissue (n = 4), and bronchoalveolar lavage fluid (BALF) (n = 1). Although clinical sensitivity of the real-time PCR assay was insufficiently sensitive (33%), the nested real-time PCR assay increased the clinical sensitivity (77%), suggesting it has a potential to be a useful method for detecting H. capsulatum DNA in clinical specimens. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Influence of human development and predators on nest survival of tundra birds, Arctic Coastal Plain, Alaska.

    PubMed

    Liebezeit, J R; Kendall, S J; Brown, S; Johnson, C B; Martin, P; McDonald, T L; Payer, D C; Rea, C L; Streever, B; Wildman, A M; Zack, S

    2009-09-01

    Nest predation may influence population dynamics of birds on the Arctic Coastal Plain (ACP) of Alaska, USA. Anthropogenic development on the ACP is increasing, which may attract nest predators by providing artificial sources of food, perches, den sites, and nest sites. Enhanced populations or concentrations of human-subsidized predators may reduce nest survival for tundra-nesting birds. In this study, we tested the hypothesis that nest survival decreases in proximity to human infrastructure. We monitored 1257 nests of 13 shorebird species and 619 nests of four passerine species at seven sites on the ACP from 2002 to 2005. Study sites were chosen to represent a range of distances to infrastructure from 100 m to 80 km. We used Cox proportional hazards regression models to evaluate the effects of background (i.e., natural) factors and infrastructure on nest survival. We documented high spatial and temporal variability in nest survival, and site and year were both included in the best background model. We did not detect an effect of human infrastructure on nest survival for shorebirds as a group. In contrast, we found evidence that risk of predation for passerine nests increased within 5 km of infrastructure. This finding provides quantitative evidence of a relationship between infrastructure and nest survival for breeding passerines on the ACP. A posteriori finer-scale analyses (within oil field sites and individual species) suggested that Red and Red-necked Phalaropes combined (Phalaropus fulicarius, P. lobatus) had lower productivity closer to infrastructure and in areas with higher abundance of subsidized predators. However, we did not detect such a relationship between infrastructure and nest survival for Semipalmated and Pectoral Sandpipers (Calidris pusilla, C. melanotos), the two most abundant shorebirds. High variability in environmental conditions, nest survival, and predator numbers between sites and years may have contributed to these inconsistent results. We recommend targeted management actions to minimize anthropogenic effects and suggest new research needed on this issue as expanding development is planned for the ACP of Alaska. In particular, we recommend research on demography of key predators and their importance with respect to nest survival, and experimental studies that better address challenges posed by high natural variability.

  3. [Contribution of nested PCR in the diagnosis of imported malaria in southern Algeria].

    PubMed

    Bouiba, L; Gassen, B; Gasmi, M; Hammadi, D; Harrat, Z

    2016-12-01

    The nested PCR was used to estimate its inputs in malaria diagnosis and in the performance of the microscope operators involved in the surveillance of malaria in remote areas of South Algeria. For the period 2010 to 2015, 112 patients (93 febrile and 19 asymptomatic) coming from sub-Saharan Africa were tested for malaria in the hospital of Tamanrasset. One part of the blood taken from fingertip was used for blood smears and the second part was absorbed in filter paper for molecular diagnosis. Overall, the infection was detected by nested PCR in 63 samples versus 53 by direct examination. In addition, 11 mixed infections and 6 positive asymptomatic cases not detected by microscopy were diagnosed by PCR. Moreover, two negative samples in nested PCR were tested positive by direct examination. The molecular tool is more sensitive than the direct examination in detecting infra-microscopic parasitaemia and mixed infections...

  4. An analysis of travel costs on transport of load and nest building in golden hamster.

    PubMed

    Guerra, Rogerio F.; Ades, Cesar

    2002-03-28

    We investigated the effects of travel costs on transporting nest material and nest-building activity in golden hamsters. Nest-deprived animals were submitted to run alleys 30, 90 and 180 cm long to access a source containing paper strips as nest material (Experiment 1) or were submitted to the same travel costs in 24-h experimental sessions (Experiment 2). We noted that increased travel costs were related to a decreased number of trips to the source, larger amounts (cm(2)) of nest material transported per trip (although total loads also decreased in longer alleys), longer intervals between trips, and increased time spent at the source and in nest building activity. Foraging efficiency (i.e. size of load divided by the time spent at the source) decreased as a function of travel costs, and animals transported their loads in two fundamental ways: in 30-cm alleys, they simply used their mouth to pull the paper strips, but in 90- or 180-cm alleys they transported the loads in their cheek pouches. The animals were faster when returning to the home-cage and their running speed (cm/s) increased as a function of the length of the alley, showing that animals are under different environmental pressures when searching for resources and subsequently running back with the load to the nest. Both male and female subjects were sensitive to travel costs, but males engaged in nest building activity more promptly and exhibited higher mean performances in most measures. We conclude that nest material is a good reinforcer, and our major results are in accordance with the predictions of microeconomic and optimal foraging theories.

  5. Effects of ketoprofen, morphine, and kappa opioids on pain-related depression of nesting in mice.

    PubMed

    Negus, S Stevens; Neddenriep, Bradley; Altarifi, Ahmad A; Carroll, F Ivy; Leitl, Michael D; Miller, Laurence L

    2015-06-01

    Pain-related functional impairment and behavioral depression are diagnostic indicators of pain and targets for its treatment. Nesting is an innate behavior in mice that may be sensitive to pain manipulations and responsive to analgesics. The goal of this study was to develop and validate a procedure for evaluation of pain-related depression of nesting in mice. Male ICR mice were individually housed and tested in their home cages. On test days, a 5- × 5-cm Nestlet was subdivided into 6 pieces, the pieces were evenly distributed on the cage floor, and Nestlet consolidation was quantified during 100-minute sessions. Baseline nesting was stable within and between subjects, and nesting was depressed by 2 commonly used inflammatory pain stimuli (intraperitoneal injection of dilute acid; intraplantar injection of complete Freund adjuvant). Pain-related depression of nesting was alleviated by drugs from 2 classes of clinically effective analgesics (the nonsteroidal anti-inflammatory drug ketoprofen and the μ-opioid receptor agonist morphine) but not by a drug from a class that has failed to yield effective analgesics (the centrally acting kappa opioid agonist U69,593). Neither ketoprofen nor morphine alleviated depression of nesting by U69,593, which suggests that ketoprofen and morphine effects were selective for pain-related depression of nesting. In contrast to ketoprofen and morphine, the kappa opioid receptor antagonist JDTic blocked depression of nesting by U69,593 but not by acid or complete Freund adjuvant. These results support utility of this procedure to assess expression and treatment of pain-related depression in mice.

  6. Stick-Nest Rat Middens as Sources of Paleoecological Data in Australian Deserts

    NASA Astrophysics Data System (ADS)

    Pearson, Stuart; Dodson, John R.

    1993-05-01

    The existing paleoenvironmental data from the Australian arid zone lack sensitivity and come from only a few sites. Macrofossils and pollen from four dated middens of the stick-nest rat ( Leporillus spp.) were analyzed from two sites in Western Australia. Animal and plant macrofossil remains were well preserved and provided evidence of change in species distribution within the last 1150 yr. Brush-tail possum and golden bandicoot have contracted their ranges in the recent past, possibly since the introduction of cats into Australia. An undescribed lacewing was also a significant find. Pollen preserved in parts of the same midden and in middens from different sites indicates that records are sensitive to the composition of the local vegetation when the midden was built. Pollen spectra are quite different from playa lakes, which record largely regional vegetation. Pollen preserved in the fecal pellets, desiccated urine, and grass mat nesting material provided similar information but some differences were apparent, suggesting dietary preferences were reflected in the fecal component. The pollen record suggested a trend to less-wooded vegetation cover in central Australia between 900 and 300 yr B.P.

  7. Development and evaluation of a 28S rRNA gene-based nested PCR assay for P. falciparum and P. vivax

    PubMed Central

    Pakalapati, Deepak; Garg, Shilpi; Middha, Sheetal; Acharya, Jyoti; Subudhi, Amit K; Boopathi, Arunachalam P; Saxena, Vishal; Kochar, Sanjay K; Kochar, Dhanpat K; Das, Ashis

    2013-01-01

    The 28S rRNA gene was amplified and sequenced from P. falciparum and P. vivax isolates collected from northwest India. Based upon the sequence diversity of the Plasmodium 28SrRNA gene in comparison with its human counterpart, various nested polymerase chain reaction (PCR) primers were designed from the 3R region of the 28SrRNA gene and evaluated on field isolates. This is the first report demonstrating the utility of this gene for species-specific diagnosis of malaria for these two species, prevalent in India. The initial evaluation on 363 clinical isolates indicated that, in comparison with microscopy, which showed sensitivity and specificity of 85.39% and 100% respectively, the sensitivity and specificity of the nested PCR assay was found to be 99.08% and 100% respectively. This assay was also successful in detecting mixed infections that are undetected by microscopy. Our results demonstrate the utility of the 28S rRNA gene as a diagnostic target for the detection of the major plasmodial species infecting humans. PMID:23816509

  8. Nest predators of open and cavity nesting birds in oak woodlands

    Treesearch

    Kathryn L. Purcell; Jared Verner

    1999-01-01

    Camera setups revealed at least three species of rodents and seven species of birds as potential predators at artificial open nests. Surprisingly, among avian predators identified at open nests, one third were Bullock's Orioles (Icterus bullockii). Two rodent species and three bird species were potential predators at artificial cavity nests. This high predator...

  9. CpG Methylation Analysis of HPV16 in Laser Capture Microdissected Archival Tissue and Whole Tissue Sections from High Grade Anal Squamous Intraepithelial Lesions: A Potential Disease Biomarker

    PubMed Central

    Molano, Monica; Tabrizi, Sepehr N.; Garland, Suzanne M.; Roberts, Jennifer M.; Machalek, Dorothy A.; Phillips, Samuel; Chandler, David; Hillman, Richard J.; Grulich, Andrew E.; Jin, Fengyi; Poynten, I. Mary; Templeton, David J.; Cornall, Alyssa M.

    2016-01-01

    Incidence and mortality rates of anal cancer are increasing globally. More than 90% of anal squamous cell carcinomas (ASCC) are associated with human papillomavirus (HPV). Studies on HPV-related anogenital lesions have shown that patterns of methylation of viral and cellular DNA targets could potentially be developed as disease biomarkers. Lesion-specific DNA isolated from formalin-fixed paraffin-embedded (FFPE) tissues from existing or prospective patient cohorts may constitute a valuable resource for methylation analysis. However, low concentrations of DNA make these samples technically challenging to analyse using existing methods. We therefore set out to develop a sensitive and reproducible nested PCR-pyrosequencing based method to accurately quantify methylation at 10 CpG sites within the E2BS1, E2BS2,3,4 and Sp1 binding sites in the viral upstream regulatory region of HPV16 genome. Methylation analyses using primary and nested PCR-pyrosequencing on 52 FFPE tissue [26 paired whole tissue sections (WTS) and laser capture microdissected (LCM) tissues] from patients with anal squamous intraepithelial lesions was performed. Using nested PCR, methylation results were obtained for the E2BS1, E2BS2,3,4 and Sp1 binding sites in 86.4% of the WTS and 81.8% of the LCM samples. Methylation patterns were strongly correlated within median values of matched pairs of WTS and LCM sections, but overall methylation was higher in LCM samples at different CpG sites. High grade lesions showed low methylation levels in the E2BS1 and E2BS2 regions, with increased methylation detected in the E2BS,3,4/Sp1 regions, showing the highest methylation at CpG site 37. The method developed is highly sensitive in samples with low amounts of DNA and demonstrated to be suitable for archival samples. Our data shows a possible role of specific methylation in the HPV16 URR for detection of HSIL. PMID:27529629

  10. CpG Methylation Analysis of HPV16 in Laser Capture Microdissected Archival Tissue and Whole Tissue Sections from High Grade Anal Squamous Intraepithelial Lesions: A Potential Disease Biomarker.

    PubMed

    Molano, Monica; Tabrizi, Sepehr N; Garland, Suzanne M; Roberts, Jennifer M; Machalek, Dorothy A; Phillips, Samuel; Chandler, David; Hillman, Richard J; Grulich, Andrew E; Jin, Fengyi; Poynten, I Mary; Templeton, David J; Cornall, Alyssa M

    2016-01-01

    Incidence and mortality rates of anal cancer are increasing globally. More than 90% of anal squamous cell carcinomas (ASCC) are associated with human papillomavirus (HPV). Studies on HPV-related anogenital lesions have shown that patterns of methylation of viral and cellular DNA targets could potentially be developed as disease biomarkers. Lesion-specific DNA isolated from formalin-fixed paraffin-embedded (FFPE) tissues from existing or prospective patient cohorts may constitute a valuable resource for methylation analysis. However, low concentrations of DNA make these samples technically challenging to analyse using existing methods. We therefore set out to develop a sensitive and reproducible nested PCR-pyrosequencing based method to accurately quantify methylation at 10 CpG sites within the E2BS1, E2BS2,3,4 and Sp1 binding sites in the viral upstream regulatory region of HPV16 genome. Methylation analyses using primary and nested PCR-pyrosequencing on 52 FFPE tissue [26 paired whole tissue sections (WTS) and laser capture microdissected (LCM) tissues] from patients with anal squamous intraepithelial lesions was performed. Using nested PCR, methylation results were obtained for the E2BS1, E2BS2,3,4 and Sp1 binding sites in 86.4% of the WTS and 81.8% of the LCM samples. Methylation patterns were strongly correlated within median values of matched pairs of WTS and LCM sections, but overall methylation was higher in LCM samples at different CpG sites. High grade lesions showed low methylation levels in the E2BS1 and E2BS2 regions, with increased methylation detected in the E2BS,3,4/Sp1 regions, showing the highest methylation at CpG site 37. The method developed is highly sensitive in samples with low amounts of DNA and demonstrated to be suitable for archival samples. Our data shows a possible role of specific methylation in the HPV16 URR for detection of HSIL.

  11. The implications of variable remigration intervals for the assessment of population size in marine turtles.

    PubMed

    Hays, G C

    2000-09-21

    Sea turtles nest on sandy beaches and tend to show high fidelity to specific nesting areas, but, despite this fidelity, the inter-annual variation in nesting numbers may be large. This variation may reflect the fact that turtles do not usually nest in consecutive years. Here, theoretical models are developed in which the interval between successive nesting years (the remigration interval) reflects conditions encountered on the feeding grounds, with good feeding years leading to a reduction in the remigration interval and vice versa. These simple models produce high levels of inter-annual variation in nesting numbers with, on occasion, almost no turtles nesting in some years even when the population is large and stable. The implications for assessing the size of sea turtle populations are considered. Copyright 2000 Academic Press.

  12. Effects of neonicotinoid insecticide exposure and monofloral diet on nest-founding bumblebee queens.

    PubMed

    Leza, Mar; Watrous, Kristal M; Bratu, Jade; Woodard, S Hollis

    2018-06-13

    Bumblebees are among the world's most important groups of pollinating insects in natural and agricultural ecosystems. Each spring, queen bumblebees emerge from overwintering and initiate new nests, which ultimately give rise to workers and new reproductives later in the season. Nest initiation and survival are thus key drivers of both bumblebee pollination services and population dynamics. We performed the first laboratory experiment with the model bumblebee species Bombus impatiens that explores how early nesting success is impacted by the effects of temporary or more sustained exposure to sublethal levels of a neonicotinoid-type insecticide (imidacloprid at 5 ppb in nectar) and by reliance on a monofloral pollen diet, two factors that have been previously implicated in bumblebee decline. We found that queens exhibited increased mortality and dramatically reduced activity levels when exposed to imidacloprid, as well as delayed nest initiation and lower brood numbers in the nest, but partially recovered from these effects when they only received early, temporary exposure. The effects of pollen diet on individual queen- and colony-level responses were overshadowed by effects of the insecticide, although a monofloral pollen diet alone was sufficient to negatively impact brood production. These findings speak to the sensitivity of queen bumblebees during the nest initiation phase of the colony cycle, with implications for how queens and their young nests are uniquely impacted by exposure to threats such as pesticide exposure and foraging habitat unsuitability. © 2018 The Author(s).

  13. Assessment of a Pan-Dermatophyte Nested-PCR Compared with Conventional Methods for Direct Detection and Identification of Dermatophytosis Agents in Animals.

    PubMed

    Piri, Fahimeh; Zarei Mahmoudabadi, Ali; Ronagh, Ali; Ahmadi, Bahram; Makimura, Koichi; Rezaei-Matehkolaei, Ali

    2018-06-26

    Conventional direct microscopy with potassium hydroxide (KOH) and culture were found to lack the ability to establish a fast and specific diagnosis of dermatophytosis. A pan-dermatophyte nested-PCR assay was developed using a novel primer pair targeting the translation elongation factor 1-α (Tef-1α) sequences for direct detection and identification of most veterinary relevant dermatophytes in animal samples suspected to dermatophytosis. A total of 140 animal skin and hair samples were subjected to direct microscopy, culture, and ITS-RFLP/ITS-sequencing of culture isolates for the detection and identification of dermatophytosis agents. Nested-PCR sequencing was performed on all the extracted DNAs using a commercial kit after dissolving the specimens by mechanical beating. Nested-PCR was positive in 90% of samples, followed by direct microscopy (85.7%) and culture (75%). The degree of agreement between nested-PCR and direct microscopy (94.4%) was higher than with culture (83.3%). In 105 culture positive cases, the measures of agreement for the identification of dermatophytosis agents were as follows: 100% between nested-PCR sequencing and ITS-RFLP/ITS-sequencing and 63.8% between nested-PCR sequencing and culture. The developed nested-PCR was faster as well as more sensitive and specific than conventional methods for detection and identification of dermatophytes in clinical samples, which was particularly suitable for epidemiological studies. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  14. Performance of a new gelled nested PCR test for the diagnosis of imported malaria: comparison with microscopy, rapid diagnostic test, and real-time PCR.

    PubMed

    Iglesias, Nuria; Subirats, Mercedes; Trevisi, Patricia; Ramírez-Olivencia, Germán; Castán, Pablo; Puente, Sabino; Toro, Carlos

    2014-07-01

    Microscopy and rapid diagnostic tests (RDTs) are the techniques commonly used for malaria diagnosis but they are usually insensitive at very low levels of parasitemia. Nested PCR is commonly used as a reference technique in the diagnosis of malaria due to its high sensitivity and specificity. However, it is a cumbersome assay only available in reference centers. We evaluated a new nested PCR-based assay, BIOMALAR kit (Biotools B&M Labs, Madrid, Spain) which employs ready-to-use gelled reagents and allows the identification of the main four species of Plasmodium. Blood samples were obtained from patients with clinical suspicion of malaria. A total of 94 subjects were studied. Fifty-two (55.3%) of them were malaria-infected subjects corresponding to 48 cases of Plasmodium falciparum, 1 Plasmodium malariae, 2 Plasmodium vivax, and 1 Plasmodium ovale. The performance of the BIOMALAR test was compared with microscopy, rapid diagnostic test (RDT) (BinaxNOW® Malaria) and real-time quantitative PCR (qPCR). The BIOMALAR test showed a sensitivity of 98.1% (95% confidence interval [CI], 89.7-100), superior to microscopy (82.7% [95% CI, 69.7-91.8]) and RDT (94.2% [95% CI, 84.1-98.8]) and similar to qPCR (100% [95% CI, 93.2-100]). In terms of specificity, the BIOMALAR assay showed the same value as microscopy and qPCR (100% [95% CI, 93.2-100]). Nine subjects were submicroscopic carriers of malaria. The BIOMALAR test identified almost all of them (8/9) in comparison with RDT (6/9) and microscopy (0/9). In conclusion, the BIOMALAR is a PCR-based assay easy to use with an excellent performance and especially useful for diagnosis submicroscopic malaria.

  15. A Molecular Approach to Nested RT-PCR Using a New Set of Primers for the Detection of the Human Immunodeficiency Virus Protease Gene.

    PubMed

    Zarei, Mohammad; Ravanshad, Mehrdad; Bagban, Ashraf; Fallahi, Shahab

    2016-07-01

    The human immunodeficiency virus (HIV-1) is the etiologic agent of AIDS. The disease can be transmitted via blood in the window period prior to the development of antibodies to the disease. Thus, an appropriate method for the detection of HIV-1 during this window period is very important. This descriptive study proposes a sensitive, efficient, inexpensive, and easy method to detect HIV-1. In this study 25 serum samples of patients under treatment and also 10 positive and 10 negative control samples were studied. Twenty-five blood samples were obtained from HIV-1-infected individuals who were receiving treatment at the acquired immune deficiency syndrome (AIDS) research center of Imam Khomeini hospital in Tehran. The identification of HIV-1-positive samples was done by using reverse transcription to produce copy deoxyribonucleic acid (cDNA) and then optimizing the nested polymerase chain reaction (PCR) method. Two pairs of primers were then designed specifically for the protease gene fragment of the nested real time-PCR (RT-PCR) samples. Electrophoresis was used to examine the PCR products. The results were analyzed using statistical tests, including Fisher's exact test, and SPSS17 software. The 325 bp band of the protease gene was observed in all the positive control samples and in none of the negative control samples. The proposed method correctly identified HIV-1 in 23 of the 25 samples. These results suggest that, in comparison with viral cultures, antibody detection by enzyme linked immunosorbent assay (ELISAs), and conventional PCR methods, the proposed method has high sensitivity and specificity for the detection of HIV-1.

  16. Network-based model of the growth of termite nests

    NASA Astrophysics Data System (ADS)

    Eom, Young-Ho; Perna, Andrea; Fortunato, Santo; Darrouzet, Eric; Theraulaz, Guy; Jost, Christian

    2015-12-01

    We present a model for the growth of the transportation network inside nests of the social insect subfamily Termitinae (Isoptera, termitidae). These nests consist of large chambers (nodes) connected by tunnels (edges). The model based on the empirical analysis of the real nest networks combined with pruning (edge removal, either random or weighted by betweenness centrality) and a memory effect (preferential growth from the latest added chambers) successfully predicts emergent nest properties (degree distribution, size of the largest connected component, average path lengths, backbone link ratios, and local graph redundancy). The two pruning alternatives can be associated with different genuses in the subfamily. A sensitivity analysis on the pruning and memory parameters indicates that Termitinae networks favor fast internal transportation over efficient defense strategies against ant predators. Our results provide an example of how complex network organization and efficient network properties can be generated from simple building rules based on local interactions and contribute to our understanding of the mechanisms that come into play for the formation of termite networks and of biological transportation networks in general.

  17. Predation of Songbird Nests Differs By Predator and Between Field and Forest Habitats

    Treesearch

    Frank R., III Thompson; Dirk E. Burhans

    2003-01-01

    Our understanding of factors affecting nest predation and ability to mitigate high nest predation rates is hampered by a lack of information on the importance of various nest predator species in different habitats and landscapes. We identified predators of songbird nests in old-field and forest habitats in central Missouri, USA, with miniature video cameras. We used an...

  18. Site fidelity, mate fidelity, and breeding dispersal in American kestrels

    USGS Publications Warehouse

    Steenhof, K.; Peterson, B.E.

    2009-01-01

    We assessed mate fidelity, nest-box fidelity, and breeding dispersal distances of American Kestrels (falco sparverius) nesting in boxes in southwestern Idaho from 1990 through 2006. Seventy-seven percent of boxes had different males and 87% had different females where nest-box occupants were identified in consecutive years. High turnover rates were partly a result of box-switching. Forty-eight percent of males and 58% of females that nested within the study area in successive years used different boxes. The probability of changing boxes was unrelated to gender, nesting success in the prior year, or years of nesting experience. Breeding dispersal distances for birds that moved to different boxes averaged 2.2 km for males (max = 22 km) and 3.2 km for females (max = 32 km). Approximately 70% of birds that nested in consecutive years on the study area had a different mate in the second year. Mate fidelity was related to box fidelity but not to prior nesting success or years of nesting experience. Mate changes occurred 32% of the time when the previous mate was known to be alive and nesting in the area. Kestrels that switched mates and boxes did not improve or decrease their subsequent nesting success. Kestrels usually switched to mates with less experience and lower lifetime productivity than their previous mates. The costs of switching boxes and mates were low, and there were no obvious benefits to fidelity. The cost of "waiting" for a previous mate that might have died could be high in species with high annual mortality.

  19. Determinants of stingless bee nest density in lowland dipterocarp forests of Sabah, Malaysia.

    PubMed

    Eltz, Thomas; Brühl, Carsten A; van der Kaars, Sander; Linsenmair, Eduard K

    2002-03-01

    We measured the nest density of stingless bees (Apidae, Meliponini) in undisturbed and logged-over dipterocarp forests in Sabah, northern Borneo, and evaluated hypotheses on proximate factors leading to the observed variation: population control mediated by (1) nest predation, (2) limitation of nest trees, or (3) food limitation. Per-area nest density varied twentyfold across 14 forest sites and was significantly affected by locality, but not by the degree and history of disturbance. Nest density was generally high in sites located in the Sepilok Forest fragment (mean 8.4 nests/ha), bordering mangroves or plantations. In contrast, nest densities in continuous forests were all low (between 0 and 2.1 nests/ha, mean 0.5 nests/ha). Yearly nest mortality was low (13.5-15.0%) over 4 years of observation and did not vary between forest localities, thus limiting the potential of nest predation (1) in creating the observed variation in nest density. The presence of potential nest trees (2), though positively correlated with nest density, explained only a minute fraction of the observed variation. Nest density was best explained by differences in the pollen resources (3) available to the bees (quantified by analysis of pollen in bee garbage). Across five selected sites the amount of nonforest pollen (from mangrove or crop plants) included in diets of Trigona collina was positively correlated with T. collina nest density. External pollen sources are a likely supplement to bee diets at times when little flowering occurs inside the forest, thus increasing overall bee carrying capacity. Pollen limitation was also indicated by direct measurements of pollen import and foraging activity of T. collina in three selected sites: Pollen traps installed at nests in high-density Sepilok captured significantly more corbicular pollen than colonies in low-density Deramakot. At the same time, morning foraging activity was also greater in Sepilok, indicating a regulatory increase in foraging in response to high pollen availability. We conclude that the abundance of stingless bees in forests in Sabah is chiefly dependent on the local availability of food resources. Bee populations strongly benefit from edge effects and increased foraging habitat diversity. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00442-001-0848-6.

  20. Nest temperatures in a loggerhead nesting beach in Turkey is more determined by sea surface than air temperature.

    PubMed

    Girondot, Marc; Kaska, Yakup

    2015-01-01

    While climate change is now fully recognised as a reality, its impact on biodiversity is still not completely understood. To predict its impact, proxies coherent with the studied ecosystem or species are thus required. Marine turtles are threatened worldwide (though some populations are recovering) as they are particularly sensitive to temperature throughout their entire life cycle. This is especially true at the embryo stage when temperature affects both growth rates and sex determination. Nest temperature is thus of prime importance to understand the persistence of populations in the context of climate change. We analysed the nest temperature of 21 loggerheads (Caretta caretta) originating from Dalyan Beach in Turkey using day-lagged generalised mixed models with autocorrelation. Surprisingly, the selected model for nest temperature includes an effect for sea surface temperature 4-times higher than for air temperature. We also detected a very significant effect of metabolic heating during development compatible with what is already known about marine turtle nests. Our new methodology allows the prediction of marine turtle nest temperatures with good precision based on a combination of air temperature measured at beach level and sea surface temperature in front of the beach. These data are available in public databases for most of the beaches worldwide. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Factors influencing depredation of artificial duck nests

    USGS Publications Warehouse

    Esler, Daniel N.; Grand, James B.

    1993-01-01

    Because artificial nests can facilitate controlled experiments of nest success, we used them to assess whether human visitation, nest density, vegetation structure, and proximity to habitat edge could affect depredation of duck nests on Yukon Flats National Wildlife Refuge, Alaska. More (P < 0.01) nests in a plot visited daily (100%) were depredated than those in plots visited at intervals of 7 (40%), 14 (35%), or 28 days (45%). More (P < 0.01) nests were depredated in a plot with 10 nests/ha (95%) than nests in a plot of a lower density (2/ha; 40%). Vegetation height, vegetation density, distance to a wetland, distance to forest edge, or distance to the nearest ecotone did not differ (P > 0.05) between depredated and undisturbed nests. We suggest that daily visitation of duck nests increases depredation, but longer intervals, typical of most nest studies, do not. High nesting densities, which could occur when flooding limits nesting habitat, may result in higher depredation rates.

  2. A semi-nested real-time PCR method to detect low chimerism percentage in small quantity of hematopoietic stem cell transplant DNA samples.

    PubMed

    Aloisio, Michelangelo; Bortot, Barbara; Gandin, Ilaria; Severini, Giovanni Maria; Athanasakis, Emmanouil

    2017-02-01

    Chimerism status evaluation of post-allogeneic hematopoietic stem cell transplantation samples is essential to predict post-transplant relapse. The most commonly used technique capable of detecting small increments of chimerism is quantitative real-time PCR. Although this method is already used in several laboratories, previously described protocols often lack sensitivity and the amount of the DNA required for each chimerism analysis is too high. In the present study, we compared a novel semi-nested allele-specific real-time PCR (sNAS-qPCR) protocol with our in-house standard allele-specific real-time PCR (gAS-qPCR) protocol. We selected two genetic markers and analyzed technical parameters (slope, y-intercept, R2, and standard deviation) useful to determine the performances of the two protocols. The sNAS-qPCR protocol showed better sensitivity and precision. Moreover, the sNAS-qPCR protocol requires, as input, only 10 ng of DNA, which is at least 10-fold less than the gAS-qPCR protocols described in the literature. Finally, the proposed sNAS-qPCR protocol could prove very useful for performing chimerism analysis with a small amount of DNA, as in the case of blood cell subsets.

  3. FilmArray, an Automated Nested Multiplex PCR System for Multi-Pathogen Detection: Development and Application to Respiratory Tract Infection

    PubMed Central

    Poritz, Mark A.; Blaschke, Anne J.; Byington, Carrie L.; Meyers, Lindsay; Nilsson, Kody; Jones, David E.; Thatcher, Stephanie A.; Robbins, Thomas; Lingenfelter, Beth; Amiott, Elizabeth; Herbener, Amy; Daly, Judy; Dobrowolski, Steven F.; Teng, David H. -F.; Ririe, Kirk M.

    2011-01-01

    The ideal clinical diagnostic system should deliver rapid, sensitive, specific and reproducible results while minimizing the requirements for specialized laboratory facilities and skilled technicians. We describe an integrated diagnostic platform, the “FilmArray”, which fully automates the detection and identification of multiple organisms from a single sample in about one hour. An unprocessed biologic/clinical sample is subjected to nucleic acid purification, reverse transcription, a high-order nested multiplex polymerase chain reaction and amplicon melt curve analysis. Biochemical reactions are enclosed in a disposable pouch, minimizing the PCR contamination risk. FilmArray has the potential to detect greater than 100 different nucleic acid targets at one time. These features make the system well-suited for molecular detection of infectious agents. Validation of the FilmArray technology was achieved through development of a panel of assays capable of identifying 21 common viral and bacterial respiratory pathogens. Initial testing of the system using both cultured organisms and clinical nasal aspirates obtained from children demonstrated an analytical and clinical sensitivity and specificity comparable to existing diagnostic platforms. We demonstrate that automated identification of pathogens from their corresponding target amplicon(s) can be accomplished by analysis of the DNA melting curve of the amplicon. PMID:22039434

  4. Procedures for offline grid nesting in regional ocean models

    NASA Astrophysics Data System (ADS)

    Mason, Evan; Molemaker, Jeroen; Shchepetkin, Alexander F.; Colas, Francois; McWilliams, James C.; Sangrà, Pablo

    One-way offline nesting of a primitive-equation regional ocean numerical model (ROMS) is investigated, with special attention to the boundary forcing file creation process. The model has a modified open boundary condition which minimises false wave reflections, and is optimised to utilise high-frequency boundary updates. The model configuration features a previously computed solution which supplies boundary forcing data to an interior domain with an increased grid resolution. At the open boundaries of the interior grid (the child) the topography is matched to that of the outer grid (the parent), over a narrow transition region. A correction is applied to the normal baroclinic and barotropic velocities at the open boundaries of the child to ensure volume conservation. It is shown that these steps, together with a carefully constructed interpolation of the parent data, lead to a high-quality child solution, with minimal artifacts such as persistent rim currents and wave reflections at the boundaries. Sensitivity experiments provide information about the robustness of the model open boundary condition to perturbations in the surface wind stress forcing field, to the perturbation of the volume conservation enforcement in the boundary forcing, and to perturbation of the vertical density structure in the boundary forcing. This knowledge is important when extending the nesting technique to include external data from alien sources, such as ocean models with physics and/or numerics different from ROMS, or from observed climatologies of temperature, salinity and sea level.

  5. High duck nesting success in a predator-reduced environment

    USGS Publications Warehouse

    Duebbert, H.F.; Lokemoen, J.T.

    1980-01-01

    Duck nesting and production were studied during 1969-74 on a 51-ha field of undisturbed grass-legume cover and a surrounding 8.13-km2 area in north-central South Dakota. The principal mammalian predators of ducks were reduced within a 259-km2 zone from May 1969 through August 1971. Dabbling duck nest densities, hatching success, and breeding populations attained high levels. Seven duck species produced 1,062 nests on the 51-ha field during 6 years, 864 (81%) hatched, 146 (14%) were destroyed, and 52 (5%) had other fates. During 1970-72, when predator reduction was most effective, the hatching success for 756 nests was 94%. The number of mallard (Anas platyrhynchos) nests increased from 37 (0.7/ha) in 1969 to 181 (3.5/ha) in 1972. Mallard pairs increased from 2.8/km2 to 16.8/km2 on the 8.13-km2 area during the same period. A minimum of 7,250 ducklings hatched on the 51-ha field during the 6 years, including 2,342 ducklings in 1972. Exceptionally high duck nesting densities and hatching rates occurred when predators were controlled.

  6. Comparison of ELISA, nested PCR and sequencing and a novel qPCR for detection of Giardia isolates from Jordan.

    PubMed

    Hijjawi, Nawal; Yang, Rongchang; Hatmal, Ma'mon; Yassin, Yasmeen; Mharib, Taghrid; Mukbel, Rami; Mahmoud, Sameer Alhaj; Al-Shudifat, Abdel-Ellah; Ryan, Una

    2018-02-01

    Little is known about the prevalence of Giardia duodenalis in human patients in Jordan and all previous studies have used direct microscopy, which lacks sensitivity. The present study developed a novel quantitative PCR (qPCR) assay at the β-giardin (bg) locus and evaluated its use as a frontline test for the diagnosis of giardiasis in comparison with a commercially available ELISA using nested PCR and sequencing of the glutamate dehydrogenase (gdh) locus (gdh nPCR) as the gold standard. A total of 96 human faecal samples were collected from 96 patients suffering from diarrhoea from 5 regions of Jordan and were screened using the ELISA and qPCR. The analytical specificity of the bg qPCR assay revealed no cross-reactions with other genera and detected all the Giardia isolates tested. Analytical sensitivity was 1 Giardia cyst per μl of DNA extract. The overall prevalence of Giardia was 64.6%. The clinical sensitivity and specificity of the bg qPCR was 89.9% and 82.9% respectively compared to 76.5 and 68.0% for the ELISA. This study is the first to compare three different methods (ELISA, bg qPCR, nested PCR and sequencing at the gdh locus) to diagnose Jordanian patients suffering from giardiasis and to analyze their demographic data. Copyright © 2018 Elsevier Inc. All rights reserved.

  7. Clinical Evaluation of a Loop-Mediated Amplification Kit for Diagnosis of Imported Malaria

    PubMed Central

    Polley, Spencer D.; González, Iveth J.; Mohamed, Deqa; Daly, Rosemarie; Bowers, Kathy; Watson, Julie; Mewse, Emma; Armstrong, Margaret; Gray, Christen; Perkins, Mark D.; Bell, David; Kanda, Hidetoshi; Tomita, Norihiro; Kubota, Yutaka; Mori, Yasuyoshi; Chiodini, Peter L.; Sutherland, Colin J.

    2013-01-01

    Background. Diagnosis of malaria relies on parasite detection by microscopy or antigen detection; both fail to detect low-density infections. New tests providing rapid, sensitive diagnosis with minimal need for training would enhance both malaria diagnosis and malaria control activities. We determined the diagnostic accuracy of a new loop-mediated amplification (LAMP) kit in febrile returned travelers. Methods. The kit was evaluated in sequential blood samples from returned travelers sent for pathogen testing to a specialist parasitology laboratory. Microscopy was performed, and then malaria LAMP was performed using Plasmodium genus and Plasmodium falciparum–specific tests in parallel. Nested polymerase chain reaction (PCR) was performed on all samples as the reference standard. Primary outcome measures for diagnostic accuracy were sensitivity and specificity of LAMP results, compared with those of nested PCR. Results. A total of 705 samples were tested in the primary analysis. Sensitivity and specificity were 98.4% and 98.1%, respectively, for the LAMP P. falciparum primers and 97.0% and 99.2%, respectively, for the Plasmodium genus primers. Post hoc repeat PCR analysis of all 15 tests with discrepant results resolved 4 results in favor of LAMP, suggesting that the primary analysis had underestimated diagnostic accuracy. Conclusions. Malaria LAMP had a diagnostic accuracy similar to that of nested PCR, with a greatly reduced time to result, and was superior to expert microscopy. PMID:23633403

  8. Rapid and sensitive detection of Curvularia lunata associated with maize leaf spot based on its Clg2p gene using semi-nested PCR.

    PubMed

    Hou, J M; Ma, B C; Zuo, Y H; Guo, L L; Gao, S G; Wang, Y Y; Liu, T

    2013-04-01

    Curvularia lunata (Wakker) Boed, the causative agent of Curvularia leaf spot in maize, was determined according to conidiophore and conidium morphology in a previous study. In the current study, a sensitive polymerase chain reaction assay was developed for the detection of C. lunata. Two specific forward (ClgD1/ClgD2) and one reverse primers (ClgD3) were designed based on a Ras-related (Clg2p) gene. Eight C. lunata isolates that represent different virulent strains in maize, six other Curvularia spp., and 22 fungal plant pathogens were used to test the specificity of the primers. PCR amplification using ClgD1/ClgD3 as the first-round primers resulted in an 870-bp band from the C. lunata isolates. The detection sensitivity using ClgD1/ClgD3 was 100 pg of genomic DNA. In the second round of PCR, a 1 : 50 dilution of the first-round PCR products was used as a template with the ClgD2/ClgD3 primer pair, which increased the detection sensitivity to 1 fg. This semi-nested PCR procedure could also be used to detect C. lunata from infected maize leaves. The proposed PCR-based assay may be used for diagnosing and monitoring maize Curvularia leaf spot. The semi-nested PCR assay may provide researchers and laboratory technologists a tool to rapidly detect C. lunata, which causes maize Curvularia leaf spot, compared with histological examination. © 2012 The Society for Applied Microbiology.

  9. Soil attributes drive nest-site selection by the campo miner Geositta poeciloptera.

    PubMed

    Meireles, Ricardo Camargos de; Teixeira, João Paulo Gusmão; Solar, Ricardo; Vasconcelos, Bruno Nery F; Fernandes, Raphael B A; Lopes, Leonardo Esteves

    2018-01-01

    Substrate type is a key-factor in nest-site selection and nest architecture of burrowing birds. However, little is known about which factors drive nest-site selection for these species, especially in the tropics. We studied the influence of soil attributes on nest-site selection by the campo miner Geositta poeciloptera, an open grassland bird that builds its nests within soil cavities. For all nests found, we measured the depth of the nest cavity and the resistance of the soil to penetration, and identified the soil horizon in which the nest was located. In soil banks with nests, we collected soil samples for granulometric analysis around each nest cavity, while in soil banks without nests we collected these samples at random points. From 43 nests found, 86% were located in the deeper soil horizons (C-horizon), and only 14% in the shallower horizons (B-horizon). Granulometric analysis showed that the C-horizons possessed a high similar granulometric composition, with high silt and low clay contents. These characteristics are associated with a low degree of structural development of the soil, which makes it easier to excavate. Contrarily, soil resistance to penetration does not seem to be an important criterion for nest site selection, although nests in more resistant the soils tend to have shallower nest cavities. Among the soil banks analyzed, 40% of those without cavities possessed a larger proportion of B-horizon relative to the C-horizon, and their texture was more clayey. On the other hand, almost all soil banks containing nest cavities had a larger C-horizon and a silty texture, indicating that soil attributes drive nest-site selection by G. poeciloptera. Thus, we conclude that the patchy distribution of G. poeciloptera can attributed to the infrequent natural exposure of the C-horizon in the tropical region, where well developed, deep and permeable soils are more common.

  10. Soil attributes drive nest-site selection by the campo miner Geositta poeciloptera

    PubMed Central

    Teixeira, João Paulo Gusmão; Solar, Ricardo; Vasconcelos, Bruno Nery F.; Fernandes, Raphael B. A.; Lopes, Leonardo Esteves

    2018-01-01

    Substrate type is a key-factor in nest-site selection and nest architecture of burrowing birds. However, little is known about which factors drive nest-site selection for these species, especially in the tropics. We studied the influence of soil attributes on nest-site selection by the campo miner Geositta poeciloptera, an open grassland bird that builds its nests within soil cavities. For all nests found, we measured the depth of the nest cavity and the resistance of the soil to penetration, and identified the soil horizon in which the nest was located. In soil banks with nests, we collected soil samples for granulometric analysis around each nest cavity, while in soil banks without nests we collected these samples at random points. From 43 nests found, 86% were located in the deeper soil horizons (C-horizon), and only 14% in the shallower horizons (B-horizon). Granulometric analysis showed that the C-horizons possessed a high similar granulometric composition, with high silt and low clay contents. These characteristics are associated with a low degree of structural development of the soil, which makes it easier to excavate. Contrarily, soil resistance to penetration does not seem to be an important criterion for nest site selection, although nests in more resistant the soils tend to have shallower nest cavities. Among the soil banks analyzed, 40% of those without cavities possessed a larger proportion of B-horizon relative to the C-horizon, and their texture was more clayey. On the other hand, almost all soil banks containing nest cavities had a larger C-horizon and a silty texture, indicating that soil attributes drive nest-site selection by G. poeciloptera. Thus, we conclude that the patchy distribution of G. poeciloptera can attributed to the infrequent natural exposure of the C-horizon in the tropical region, where well developed, deep and permeable soils are more common. PMID:29381768

  11. Nest-site selection and nest success of an Arctic-breeding passerine, Smith's Longspur, in a changing climate

    USGS Publications Warehouse

    McFarland, Heather R.; Kendall, Steve J.; Powell, Abby

    2017-01-01

    Despite changes in shrub cover and weather patterns associated with climate change in the Arctic, little is known about the breeding requirements of most passerines tied to northern regions. We investigated the nesting biology and nest habitat characteristics of Smith's Longspurs (Calcarius pictus) in 2 study areas in the Brooks Range of Alaska, USA. First, we examined variation in nesting phenology in relation to local temperatures. We then characterized nesting habitat and analyzed nest-site selection for a subset of nests (n = 86) in comparison with paired random points. Finally, we estimated the daily survival rate of 257 nests found in 2007–2013 with respect to both habitat characteristics and weather variables. Nest initiation was delayed in years with snow events, heavy rain, and freezing temperatures early in the breeding season. Nests were typically found in open, low-shrub tundra, and never among tall shrubs (mean shrub height at nests = 26.8 ± 6.7 cm). We observed weak nest-site selection patterns. Considering the similarity between nest sites and paired random points, coupled with the unique social mating system of Smith's Longspurs, we suggest that habitat selection may occur at the neighborhood scale and not at the nest-site scale. The best approximating model explaining nest survival suggested a positive relationship with the numbers of days above 21°C that an individual nest experienced; there was little support for models containing habitat variables. The daily nest survival rate was high (0.972–0.982) compared with that of most passerines in forested or grassland habitats, but similar to that of passerines nesting on tundra. Considering their high nesting success and ability to delay nest initiation during inclement weather, Smith's Longspurs may be resilient to predicted changes in weather regimes on the breeding grounds. Thus, the greatest threat to breeding Smith's Longspurs associated with climate change may be the loss of low-shrub habitat types, which could significantly change the characteristics of breeding areas.

  12. Relative effects of plumage coloration and vegetation density on nest success

    USGS Publications Warehouse

    Miller, M.W.

    1999-01-01

    Many passerine species are highly dichromatic with brightly-colored males and cryptically-colored females. Bright plumage in males is commonly thought to arise as a result of sexual selection by females such that males with bright coloration possess high fitness. However, bright plumage potentially could expose males to increased predation risk. Consistent with this idea, males of many highly dichromatic passerine species do not incubate. I tested whether brightly-colored males avoid incubation to reduce the probability of visual predators locating their nest. This hypothesis predicts greater hatching success for clutches incubated by cryptically-colored individuals than by brightly-colored individuals. The Northern Cardinal (Cardinalis cardinalis) is a common dichromatic species that breeds throughout the eastern U.S. I placed two button-quail (Turnix st).) eggs in each of 203 simulated cardinal nests. Dull brown cardboard, simulating a female cardinal, was placed over about half of all clutches. Bright red cardboard, simulating a male cardinal, was placed over the other clutches. Nest success was highest for well-concealed nests (87%) and lowest for nests in open habitat (54%). Nests containing red cardboard did not have significantly lower success than nests with brown cardboard, nor did I detect a significant color X vegetation-density interaction. My analysis may have had insufficient power to detect an effect of color on nest success; alternatively, brightly-colored males that do not incubate may achieve benefits unrelated to predation risk.

  13. Parental investment decisions in response to ambient nest-predation risk versus actual predation on the prior nest

    USGS Publications Warehouse

    Chalfoun, A.D.; Martin, T.E.

    2010-01-01

    Theory predicts that parents should invest less in dependent offspring with lower reproductive value, such as those with a high risk of predation. Moreover, high predation risk can favor reduced parental activity when such activity attracts nest predators. Yet, the ability of parents to assess ambient nest-predation risk and respond adaptively remains unclear, especially where nest-predator assemblages are diverse and potentially difficult to assess. We tested whether variation in parental investment by a multi-brooded songbird (Brewer's Sparrow, Spizella breweri) in an environment (sagebrush steppe) with diverse predators was predicted by ambient nest-predation risk or direct experience with nest predation. Variation among eight sites in ambient nest-predation risk, assayed by daily probabilities of nest predation, was largely uncorrelated across four years. In this system risk may therefore be unpredictable, and aspects of parental investment (clutch size, egg mass, incubation rhythms, nestling-feeding rates) were not related to ambient risk. Moreover, investment at first nests that were successful did not differ from that at nests that were depredated, suggesting parents could not assess and respond to territorylevel nest-predation risk. However, parents whose nests were depredated reduced clutch sizes and activity at nests attempted later in the season by increasing the length of incubation shifts (on-bouts) and recesses (off-bouts) and decreasing trips to feed nestlings. In this unpredictable environment parent birds may therefore lack sufficient cues of ambient risk on which to base their investment decisions and instead rely on direct experience with nest predation to inform at least some of their decisions. ?? 2010 The Cooper Ornithological Society.

  14. Response of beach-nesting American Oystercatchers to off-road vehicles: An experimental approach reveals physiological nuances and decreased nest attendance

    USGS Publications Warehouse

    Felton, Shilo K.; Pollock, Kenneth H.; Simons, Theodore R.

    2018-01-01

    Shorebird populations face increasing challenges as rising sea levels and growing human populations constrain their breeding habitats. On recreational beaches, the nesting season often coincides with a season of high visitor use, increasing the potential for conflict, which may negatively influence beach-nesting shorebird species. We designed a field experiment to study the responses of nesting American Oystercatchers (Haematopus palliatus) to off-road passenger vehicles (ORVs) at Cape Hatteras and Cape Lookout National Seashores in North Carolina, USA. We used continuous video and heart rate recordings to assess changes in the behavior and physiology of incubating oystercatchers. We conducted driving experiments affecting 7 nesting pairs in 2014 and 19 nesting pairs in 2015, between April and July of each year. Experimental treatments were repeated throughout the incubation period for each nest. Although responses were highly variable within and among pairs, paired randomized permutation tests indicated that, overall, oystercatcher pairs spent a greater proportion of time with their heads up and exhibited slower heart rates during driving treatments. Pairs also left their nests more frequently and attended their nests for a lower proportion of time during driving treatments, although these responses diminished over time. Higher nest attendance and lower departure rates late in incubation may have reflected a stronger attachment to nests closer to hatching or habituation to the driving treatment, although individuals continued to exhibit physiological responses to passing vehicles throughout incubation. Beach-nesting birds may benefit from reduced vehicle traffic at their nesting sites, allowing parents to spend more time attending the nest and less time on defensive behaviors.

  15. Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems.

    PubMed

    Ximenes, Camila; Brandão, Eduardo; Oliveira, Paula; Rocha, Abraham; Rego, Tamisa; Medeiros, Rafael; Aguiar-Santos, Ana; Ferraz, João; Reis, Christian; Araujo, Paulo; Carvalho, Luiz; Melo, Fabio L

    2014-12-01

    The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.

  16. Teaching Ecological Concepts with Mud Dauber Nests.

    ERIC Educational Resources Information Center

    Matthews, Robert W.; Matthews, Janice R.

    1999-01-01

    Contends that mud dauber nests--which are widely available, safe, inexpensive, and easy to use--offer a novel and highly motivating way to teach ecological concepts to life science students at many grade levels. Presents background information for teachers, details classroom-tested methods for nest dissection, provides keys to nest contents, and…

  17. Using Artificial Nests to Study Nest Predation in Birds

    ERIC Educational Resources Information Center

    Belthoff, James R.

    2005-01-01

    A simple and effective field exercise that demonstrates factors affecting predation on bird nests is described. With instructor guidance, students in high school biology or college-level biology, ecology, animal behavior, wildlife management or ornithology laboratory courses can collaborate to design field experiments related to nest depredation.

  18. Changes of loggerhead turtle (Caretta caretta) dive behavior associated with tropical storm passage during the inter-nesting period.

    PubMed

    Wilson, Maria; Tucker, Anton D; Beedholm, Kristian; Mann, David A

    2017-10-01

    To improve conservation strategies for threatened sea turtles, more knowledge on their ecology, behavior, and how they cope with severe and changing weather conditions is needed. Satellite and animal motion datalogging tags were used to study the inter-nesting behavior of two female loggerhead turtles in the Gulf of Mexico, which regularly has hurricanes and tropical storms during nesting season. We contrast the behavioral patterns and swimming energetics of these two turtles, the first tracked in calm weather and the second tracked before, during and after a tropical storm. Turtle 1 was highly active and swam at the surface or submerged 95% of the time during the entire inter-nesting period, with a high estimated specific oxygen consumption rate (0.95 ml min -1  kg -0.83 ). Turtle 2 was inactive for most of the first 9 days of the inter-nesting period, during which she rested at the bottom (80% of the time) with low estimated oxygen consumption (0.62 ml min -1  kg -0.83 ). Midway through the inter-nesting period, turtle 2 encountered a tropical storm and became highly active (swimming 88% of the time during and 95% after the storm). Her oxygen consumption increased significantly to 0.97 ml min -1  kg -0.83 during and 0.98 ml min -1  kg -0.83 after the storm. However, despite the tropical storm, turtle 2 returned to the nesting beach, where she successfully re-nested 75 m from her previous nest. Thus, the tropical storm had a minor effect on this female's individual nesting success, even though the storm caused 90% loss nests at Casey Key. © 2017. Published by The Company of Biologists Ltd.

  19. Environmental modulation of androgen levels and secondary sex characters in two populations of the peacock blenny Salaria pavo.

    PubMed

    Saraiva, João L; Gonçalves, David M; Oliveira, Rui F

    2010-02-01

    Morphology and endocrinology were studied in two populations of the peacock blenny Salaria pavo, with different regimes of sexual selection imposed by differences in nest site availability. The peacock blenny is a small, sexually dimorphic benthic fish that presents exclusive paternal care of the clutch and inhabits rocky shores of the Mediterranean and adjacent Atlantic areas. In a population from the Gulf of Trieste (Northern Adriatic sea) inhabiting rocky shores where nest sites are abundant, male-male competition for nests is low, males court females and a low frequency of alternative reproductive tactics (small, parasitic female-mimicking sneaker males that change tactic into nest holders in subsequent breeding seasons) occurs. Conversely at Ria Formosa, a coastal lagoon in Southern Portugal, where nest sites are scarce and highly aggregated, male-male competition for nests is very high, there is sex-role reversal with female courtship and a high frequency of alternative reproductive tactics is observed. Concomitantly, at Ria Formosa nest holder males are larger and present more developed secondary sex characters and higher levels of 11KT than at the Gulf of Trieste. However, the gonads of nest holders and parasitic males were larger in the Gulf of Trieste population. Competition for nests at Ria Formosa seems to promote more developed secondary sex characters in nest site scarcity conditions, while competition for females at the Gulf of Trieste seems to be spurring sperm competition among males in populations where nest sites are more abundant. 11KT was thus associated with the development and expression of secondary sex characters in contrasting environments. These results exemplify how the modulation of behavioral plasticity and secondary sex characters by the social environment can be mediated by androgens. Copyright 2009 Elsevier Inc. All rights reserved.

  20. High resolution regional climate simulation of the Hawaiian Islands - Validation of the historical run from 2003 to 2012

    NASA Astrophysics Data System (ADS)

    Xue, L.; Newman, A. J.; Ikeda, K.; Rasmussen, R.; Clark, M. P.; Monaghan, A. J.

    2016-12-01

    A high-resolution (a 1.5 km grid spacing domain nested within a 4.5 km grid spacing domain) 10-year regional climate simulation over the entire Hawaiian archipelago is being conducted at the National Center for Atmospheric Research (NCAR) using the Weather Research and Forecasting (WRF) model version 3.7.1. Numerical sensitivity simulations of the Hawaiian Rainband Project (HaRP, a filed experiment from July to August in 1990) showed that the simulated precipitation properties are sensitive to initial and lateral boundary conditions, sea surface temperature (SST), land surface models, vertical resolution and cloud droplet concentration. The validations of model simulated statistics of the trade wind inversion, temperature, wind field, cloud cover, and precipitation over the islands against various observations from soundings, satellites, weather stations and rain gauges during the period from 2003 to 2012 will be presented at the meeting.

  1. Plasma steroid profiles in nesting loggerhead turtles (Caretta caretta) in Queensland, Australia: relationship to nesting episode and season.

    PubMed

    Whittier, J M; Corrie, F; Limpus, C

    1997-04-01

    Plasma levels of four hormones-progesterone (P), testosterone (T), estradiol 17-beta (E2), and corticosterone (B)-were measured in samples taken from nesting female loggerhead turtles (Caretta caretta) by using specific radioimmunoassays. Samples were taken in an early, middle, or late period during the summer nesting season from females at first, second, third, or > fourth nesting episodes, defined as successive within-season nesting events, at Mon Repos Beach, Queensland, Australia. Data on individual patterns of nesting, collected over the past 20 years by the Queensland Turtle Research Project, and the seasonal nesting data, were analyzed with respect to influences on hormonal profiles. Circulating levels of E2 were mostly undetectable, suggesting either that this estrogen is not produced at this time of nesting, or that, perhaps, another estrogen may be present that is not detected by the specific radioimmunoassay. P, T, and B profiles in the nesting females were associated with the individual turtles' progression through successive nesting episodes, with a marked decline in all three hormones by the last (> 4) nesting episode of the season. Nesting episode accounted for significant changes that were related to season, in that nesting episode and season were significantly correlated. These patterns were observed in the population, when singly sampled at each time period or nesting episode, and in individual females sampled repeatedly over time. Moreover, T and B were highly and significantly correlated in females at all nesting episodes and time periods, and in the singly and repeatedly sampled females. The magnitude of change in T and B over time was also highly and significantly correlated in repeatedly sampled females. Together these results suggest the hypothesis that T and B interact over the period of successive nesting and may be involved in reproductive functions such as the mobilization of reserves for egg production in C. caretta.

  2. Daily survival rates for nests of Black Skimmers from a core breeding area of the Southeastern USA

    USGS Publications Warehouse

    Brooks, Gillian L.; Sanders, Felicia J.; Gerard, Patrick D.; Jodice, Patrick G.R.

    2014-01-01

    Little is known about the reproductive success of Black Skimmers (Rynchops niger) throughout the southeastern USA where availability of undisturbed beaches for nesting is limited. Daily survival rates (DSR) of nests were examined at three nesting sites in Cape Romain National Wildlife Refuge (CRNWR), South Carolina, USA, 2009–2010. The percent of successful nests (n  =  346 nests) ranged from 42–69% among colony sites when data were pooled across both years. The DSR of nests was primarily related to colony site, predation risk, height of high tide, and clutch size. Predation and overwash were the principal causes of identifiable nest loss, each accounting for ∼33% of nest failures during the two study years. Because of the challenges of resighting skimmer chicks, we were not able to measure chick survival effectively and therefore accurate measures of productivity remain elusive. High variability in nest success among sites within close proximity to each other (<20 km) suggests factors at local scales such as disturbance, predation, and overwash events strongly influenced nest success of Black Skimmers during these 2 years as opposed to more region-wide stressors such as tropical storms or food availability. Although time-intensive techniques to control predators do exist, management options to limit flooding and overwash are far more limited. Conservation of Black Skimmers in the southeastern USA would benefit from coordinated, multi-state efforts to measure nest and chick survival.

  3. Inadequate thermal refuge constrains landscape habitability for a grassland bird species

    PubMed Central

    Pierce, Brian L.

    2017-01-01

    Ecologists have long recognized the influence that environmental conditions have on abundance and range extent of animal species. We used the northern bobwhite (Colinus virginianus; hereafter bobwhite) as a model species for studying how microclimates serve as refuge against severe weather conditions. This species serves as an indicator or umbrella species for other sensitive ground-nesting, grassland obligate species. We conducted a mensurative field experiment in the rolling plains of Texas, USA, a semi-arid ecosystem on the southwestern periphery of bobwhite range, to determine whether native bunch grasses, apparently suitable for bobwhite nesting, could reduce ambient temperature below levels harmful for eggs. During the nesting season, we compared temperature and relative humidity readings at daily heat maxima (i.e., the 3 h during each day with highest temperatures) during the nesting season over the course of two years at 63 suitable nest sites paired with 63 random locations (n = 126) using two sensors at ∼10 and ∼60 cm above ground level. Mean temperature at nest height was 2.3% cooler at nest sites (35.99 °C ± 0.07 SE) compared to random locations (36.81 °C ± 0.07 SE); at ambient height, nest sites were slightly cooler (32.78 °C ± 0.06 SE) than random location (32.99 °C ± 0.06 SE). Mean relative humidity at nest sites was greater at nest height (34.53% ± 0.112 SE) and ambient height (36.22% ± 0.10 SE) compared to random locations at nest (33.35% ± 0.12 SE) and ambient height (35.75% ± 0.10 SE). Based on these results, cover at sites that appear visually suitable for nesting by bobwhites and other ground nesting birds provided adequate thermal refuge in the rolling plains by maintaining cooler, moister microclimates than surrounding non-nesting locations. Post-hoc analyses of data revealed that habitat conditions surrounding suitable nest sites strongly influenced thermal suitability of the substrate. Given that eggs of bobwhites and probably other species would experience lethal temperatures without these thermal refuges in the context of proper habitat condition, nesting vegetation is a critical component of niche space for bobwhites and other ground nesting birds in semi-arid regions. Many contemporary land uses, however, degrade or destroy bunch grasses and grassland systems, and thus decrease landscape inhabitability. Conservationists working with obligate grassland species that require bunch grasses in semi-arid regions should develop land management strategies that maximize the availability of these thermal refuges across space and time. PMID:28828282

  4. Inadequate thermal refuge constrains landscape habitability for a grassland bird species.

    PubMed

    Tomecek, John M; Pierce, Brian L; Reyna, Kelly S; Peterson, Markus J

    2017-01-01

    Ecologists have long recognized the influence that environmental conditions have on abundance and range extent of animal species. We used the northern bobwhite ( Colinus virginianus ; hereafter bobwhite) as a model species for studying how microclimates serve as refuge against severe weather conditions. This species serves as an indicator or umbrella species for other sensitive ground-nesting, grassland obligate species. We conducted a mensurative field experiment in the rolling plains of Texas, USA, a semi-arid ecosystem on the southwestern periphery of bobwhite range, to determine whether native bunch grasses, apparently suitable for bobwhite nesting, could reduce ambient temperature below levels harmful for eggs. During the nesting season, we compared temperature and relative humidity readings at daily heat maxima (i.e., the 3 h during each day with highest temperatures) during the nesting season over the course of two years at 63 suitable nest sites paired with 63 random locations ( n = 126) using two sensors at ∼10 and ∼60 cm above ground level. Mean temperature at nest height was 2.3% cooler at nest sites (35.99 °C ± 0.07 SE) compared to random locations (36.81 °C ± 0.07 SE); at ambient height, nest sites were slightly cooler (32.78 °C ± 0.06 SE) than random location (32.99 °C ± 0.06 SE). Mean relative humidity at nest sites was greater at nest height (34.53% ± 0.112 SE) and ambient height (36.22% ± 0.10 SE) compared to random locations at nest (33.35% ± 0.12 SE) and ambient height (35.75% ± 0.10 SE). Based on these results, cover at sites that appear visually suitable for nesting by bobwhites and other ground nesting birds provided adequate thermal refuge in the rolling plains by maintaining cooler, moister microclimates than surrounding non-nesting locations. Post-hoc analyses of data revealed that habitat conditions surrounding suitable nest sites strongly influenced thermal suitability of the substrate. Given that eggs of bobwhites and probably other species would experience lethal temperatures without these thermal refuges in the context of proper habitat condition, nesting vegetation is a critical component of niche space for bobwhites and other ground nesting birds in semi-arid regions. Many contemporary land uses, however, degrade or destroy bunch grasses and grassland systems, and thus decrease landscape inhabitability. Conservationists working with obligate grassland species that require bunch grasses in semi-arid regions should develop land management strategies that maximize the availability of these thermal refuges across space and time.

  5. Dynamics of habitat selection in birds: adaptive response to nest predation depends on multiple factors.

    PubMed

    Devries, J H; Clark, R G; Armstrong, L M

    2018-05-01

    According to theory, habitat selection by organisms should reflect underlying habitat-specific fitness consequences and, in birds, reproductive success has a strong impact on population growth in many species. Understanding processes affecting habitat selection also is critically important for guiding conservation initiatives. Northern pintails (Anas acuta) are migratory, temperate-nesting birds that breed in greatest concentrations in the prairies of North America and their population remains below conservation goals. Habitat loss and changing land use practices may have decoupled formerly reliable fitness cues with respect to nest habitat choices. We used data from 62 waterfowl nesting study sites across prairie Canada (1997-2009) to examine nest survival, a primary fitness metric, at multiple scales, in combination with estimates of habitat selection (i.e., nests versus random points), to test for evidence of adaptive habitat choices. We used the same habitat covariates in both analyses. Pintail nest survival varied with nest initiation date, nest habitat, pintail breeding pair density, landscape composition and annual moisture. Selection of nesting habitat reflected patterns in nest survival in some cases, indicating adaptive selection, but strength of habitat selection varied seasonally and depended on population density and landscape composition. Adaptive selection was most evident late in the breeding season, at low breeding densities and in cropland-dominated landscapes. Strikingly, at high breeding density, habitat choice appears to become maladaptive relative to nest predation. At larger spatial scales, the relative availability of habitats with low versus high nest survival, and changing land use practices, may limit the reproductive potential of pintails.

  6. Sensitivity of breeding parameters to food supply in Black-legged Kittiwakes Rissa tridactyla

    USGS Publications Warehouse

    Gill, Verena A.; Hatch, Scott A.; Lanctot, Richard B.

    2002-01-01

    We fed Herring Clupea pallasi to pairs of Black-legged Kittiwakes Rissa tridactyla throughout the breeding season in two years at a colony in the northern Gulf of Alaska. We measured responses to supplemental feeding in a wide array of breeding parameters to gauge their relative sensitivity to food supply, and thus their potential as indicators of natural foraging conditions. Conventional measures of success (hatching, fledging and overall productivity) were more effective as indicators of food supply than behavioural attributes such as courtship feeding, chick provisioning rates and sibling aggression. However, behaviour such as nest relief during incubation and adult attendance with older chicks were also highly responsive to supplemental food and may be useful for monitoring environmental conditions in studies of shorter duration. On average, the chick-rearing stage contained more sensitive indicators of food availability than prelaying or incubation stages. Overall, rates of hatching and fledging success, and the mean duration of incubation shifts were the most food-sensitive parameters studied.

  7. A novel loop-mediated isothermal amplification-based test for detecting Neospora caninum DNA.

    PubMed

    Ramos, Andrea Estefanía; Muñoz, Marina; Cortés-Vecino, Jesús Alfredo; Barato, Paola; Patarroyo, Manuel Alfonso

    2017-11-29

    Neospora caninum is a cyst-forming, coccidian parasite which is known to cause neurological disorders in dogs and abortion and neonatal mortality in cows and other livestock. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay based on the Neospora caninum Nc-5 gene and compares its efficacy for detecting DNA to that of a semi-nested PCR test. Six primers were designed based on the Nc-5 repeat region of N. caninum. Specific LAMP primers led to successful amplification of N. caninum DNA at 63 °C in 30 min. The LAMP assay was highly specific (i.e. it did not reveal cross-reactivity with other parasite species) and had a low N. caninum plasmid DNA limit of detection (1 fg), which is ten times higher than that for the semi-nested PCR. LAMP applicability was evaluated using a set of naturally-infected samples (59 from canine faeces and five from bovine abortions). Thirty-nine percent (25/64) of the naturally-infected samples were positive for N. caninum DNA by LAMP and 36% (23/64) by semi-nested PCR. However, the LAMP assay is much faster to perform than semi-nested PCR and provides results in 30 min. The optimized reaction conditions described in this study resulted in a sensitive, specific and rapid technique for detecting N. caninum DNA. Considering the advantages of LAMP for detecting N. caninum DNA, further assays aimed at testing its usefulness on a wider range of field samples are recommended.

  8. Nest survival relative to patch size in a highly fragmented shortgrass prairie landscape

    USGS Publications Warehouse

    Skagen, S.K.; Yackel Adams, A.A.; Adams, R.D.

    2005-01-01

    Understanding the influences of habitat fragmentation on vertebrate populations is essential for the protection and ecological restoration of strategic sites for native species. We examined the effects of prairie fragmentation on avian reproductive success using artificial and natural nests on 26 randomly selected, privately owned patches of shortgrass prairie ranging in size from 7 to 454 ha within a cropland matrix in Washington County, Colorado, summer 2000. Survival trends of artificial and natural nests differed. Daily survival of artificial nests increased with patch size up to about 65 ha and differed little at larger patch sizes, whereas daily survival of Lark Bunting (Calamospiza melanocorys) and Horned Lark (Eremophila alpestris) nests decreased with increasing size of the grassland patch. We hypothesize that our unexpected findings of lower survival of natural nests with increasing patch sizes and different trends between artificial and natural nests are due to the particular structure of predator communities in our study area and the ways in which individual predators respond to artificial and natural nests. We recommend that the value of small habitat patches in highly fragmented landscapes not be overlooked.

  9. Effects of Invasive European Fire Ants (Myrmica rubra) on Herring Gull (Larus argentatus) Reproduction

    PubMed Central

    DeFisher, Luke E.; Bonter, David N.

    2013-01-01

    Various invasive ant species have negatively affected reproductive success in birds by disrupting nest site selection, incubation patterns, food supply, and by direct predation on nestlings. Impacts can be particularly severe when non-native ants colonize seabird nesting islands where thousands of birds may nest in high densities on the ground or in burrows or crevices. Here we report on the first documented effects of Myrmica rubra, the European fire ant, on the reproduction of birds in its non-native range. We documented herring gulls (Larus argentatus) on Appledore Island, Maine, engaging in more erratic incubation behaviors at nests infested by the ants. Newly-hatched chicks in some nests were swarmed by ants, leading to rapid chick death. Due to high overall rates of chick mortality, survival probabilities did not vary between nests with and without ant activity, however chick growth rates were slower at nests with ants than at ant-free nests. Ant infestation likely leads to longer-term fitness consequences because slower growth rates early in life may ultimately lead to lower post-fledging survival probabilities. PMID:23691168

  10. Caterpillars benefit from thermal ecosystem engineering by wandering albatrosses on sub-Antarctic Marion Island.

    PubMed

    Sinclair, Brent J; Chown, Steven L

    2006-03-22

    Wandering albatrosses (Diomedea exulans) nest on Southern Ocean islands, building elevated nests upon which they incubate eggs and raise chicks, and which the chicks occupy through winter. The nests support high invertebrate biomass, including larvae of the flightless moth Pringleophaga marioni. Here we argue that high biomass of P. marioni in the nests is not associated with nutrient loading as previously suspected, but that higher temperatures in the nests increase growth and feeding rate, and decrease deleterious repeated cold exposure, providing fitness advantages for P. marioni. Thus, wandering albatrosses may be serving as thermal engineers, modifying temperature and therefore enabling better resource use by P. marioni.

  11. The possible function of stone ramparts at the nest entrance of the blackstart.

    PubMed

    Leader; Yom-tov

    1998-07-01

    Blackstarts, Cercomela melanura, Turdidae, construct a rampart of stones at the entrance to their nests. These ramparts may reach remarkable proportions, containing hundreds of flat rocks. We investigated several hypotheses regarding the function of stone ramparts, by monitoring individually marked blackstarts at the En-Gedi Nature Reserve, Israel. Stones were collected solely by females, who carried them in their beaks, while flying to the nest, at a rate of up to one stone per min, after pair formation had occurred. The number and total weight of stones as well as rampart height showed a highly positive correlation with the size of the nest cavity opening. The rampart decreased the size of the cavity entrance to some nests by as much as 67%, which suggests an antipredator barrier function. Survival rates of eggs and chicks were extremely low and the major cause of reproductive failure was predation. Successful nests tended to be located higher off the ground than predated nests, and often contained fewer stones. Furthermore, larger females in terms of wing and tail length nested in cavities higher off the ground and built smaller ramparts containing lighter stones. An artificial nest predation experiment did not reveal a difference in predation rates between nests with and without stone ramparts. Spiny mice, Acomys sp., were the main egg predators. However, in 37% of nests with ramparts that were predated, the perpetrator flattened the rampart, suggesting that they may serve as a barricade, forcing the predator to invest time in clearing the stones in order to gain access to the nest, and perhaps allowing the nesting female sufficient time to escape. We propose, therefore, that owing to such high nest predation rates, females nesting close to the ground build stone ramparts as an 'early warning' mechanism to prevent themselves from being trapped inside nest cavities and predated. Copyright 1998 The Association for the Study of Animal Behaviour.

  12. Identifying effective actions to guide volunteer-based and nationwide conservation efforts for a ground-nesting farmland bird.

    PubMed

    Santangeli, Andrea; Arroyo, Beatriz; Millon, Alexandre; Bretagnolle, Vincent

    2015-08-01

    1. Modern farming practices threaten wildlife in different ways, and failure to identify the complexity of multiple threats acting in synergy may result in ineffective management. To protect ground-nesting birds in farmland, monitoring and mitigating impacts of mechanical harvesting is crucial. 2. Here, we use 6 years of data from a nationwide volunteer-based monitoring scheme of the Montagu's harrier, a ground-nesting raptor, in French farmlands. We assess the effectiveness of alternative nest protection measures and map their potential benefit to the species. 3. We show that unprotected nests in cultivated land are strongly negatively affected by harvesting and thus require active management. Further, we show that protection from harvesting alone (e.g. by leaving a small unharvested buffer around the nest) is impaired by post-harvest predation at nests that become highly conspicuous after harvest. Measures that simultaneously protect from harvesting and predation (by adding a fence around the nest) significantly enhance nest productivity. 4. The map of expected gain from nest protection in relation to available volunteers' workforce pinpoints large areas of high expected gain from nest protection that are not matched by equally high workforce availability. This mismatch suggests that the impact of nest protection can be further improved by increasing volunteer efforts in key areas where they are low relative to the expected gain they could have. 5. Synthesis and applications . This study shows that synergistic interplay of multiple factors (e.g. mechanical harvesting and predation) may completely undermine the success of well-intentioned conservation efforts. However, identifying areas where the greatest expected gains can be achieved relative to effort expended can minimize the risk of wasted volunteer actions. Overall, this study underscores the importance of citizen science for collecting large-scale data useful for producing science and ultimately informs large-scale evidence-based conservation actions within an adaptive management framework.

  13. Efficacy of the detection of Legionella in hot and cold water samples by culture and PCR. I. Standardization of methods.

    PubMed

    Wójcik-Fatla, Angelina; Stojek, Nimfa Maria; Dutkiewicz, Jacek

    2012-01-01

    The aim of the present study was: - to compare methods for concentration and isolation of Legionella DNA from water; - to examine the efficacy of various modifications of PCR test (PCR, semi-nested PCR, and real-time PCR) for the detection of known numbers of Legionella pneumophila in water samples artificially contaminated with the strain of this bacterium and in randomly selected samples of environmental water, in parallel with examination by culture. It was found that filtration is much more effective than centrifugation for the concentration of DNA in water samples, and that the Qiamp DNA Mini-Kit is the most efficient for isolation of Legionella DNA from water. The semi-nested PCR and real-time PCR proved to be the most sensitive methods for detection of Legionella DNA in water samples. Both PCR modifications showed a high correlation with recovery of Legionella by culture (p<0.01), while no correlation occurred between the results of one-stage PCR and culture (p>0.1).

  14. Wildlife studies on the Hanford Site: 1993 Highlights report

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cadwell, L.L.

    The Pacific Northwest Laboratory (PNL) Wildlife Resources Monitoring Project was initiated by DOE to track the status of wildlife populations to determine whether Hanford operations affected them. The project continues to conduct a census of wildlife populations that are highly visible, economically or aesthetically important, and rare or otherwise considered sensitive. Examples of long-term data collected and maintained through the Wildlife Resources Monitoring Project include annual goose nesting surveys conducted on islands in the Hanford Reach, wintering bald eagle surveys, and fall Chinook salmon redd (nest) surveys. The report highlights activities related to salmon and mollusks on the Hanford Reachmore » of the Columbia River; describes efforts to map vegetation on the Site and efforts to survey species of concern; provides descriptions of shrub-steppe bird surveys, including bald eagles, Canada geese, and hawks; outlines efforts to monitor mule deer and elk populations on the Site; and describes development of a biological database management system.« less

  15. The importance of odor in nest site selection by a lodger bee, Centris Bicornuta Mocsáry (Hymenoptera: Apidae) in the dry forest of Costa Rica.

    PubMed

    Vinson, S B; Frankie, G W; Rao, A

    2011-01-01

    The more common lodger bee occurring in the dry forest of Costa Rica, Centris bicornuta Muscáry), has been observed nesting in new nest cavities drilled into wooden blocks placed next to cavities used by another female within 2-3 days. In contrast, new nest cavities placed in similar areas with no nesting Centris nearby were not used for weeks. These observations suggest that the presence of nesting bees may play a role in nest site selection. To confirm our observations, new nest cavities were placed in areas with or without nesting. We found nest initiation in newly placed nest cavities only in areas where bees were actively nesting. To examine the possibility that nesting locations are not unique, we placed new nest cavities in new locations either with (a) a number of completed nest cavities or (b) placed alone. Within three days we only found bees nesting in the newly placed nest cavities in situation "a". The results suggested that odor might be involved. We next compared nesting in new cavities placed alone with cavities contaminated with either (a) nest entrance plug material, (b) nest nectar, (c) nest pollen or (d) a combination of pollen and nectar. Nesting was significantly low in cavities placed next to cavities with nest entrance plug material (a), and high in cavities placed next to cavities "b, c, or d". The results suggest that pollen and /or nectar odor play a role in the location of potential nest sites.

  16. Psychophysical Laws and the Superorganism.

    PubMed

    Reina, Andreagiovanni; Bose, Thomas; Trianni, Vito; Marshall, James A R

    2018-03-12

    Through theoretical analysis, we show how a superorganism may react to stimulus variations according to psychophysical laws observed in humans and other animals. We investigate an empirically-motivated honeybee house-hunting model, which describes a value-sensitive decision process over potential nest-sites, at the level of the colony. In this study, we show how colony decision time increases with the number of available nests, in agreement with the Hick-Hyman law of psychophysics, and decreases with mean nest quality, in agreement with Piéron's law. We also show that colony error rate depends on mean nest quality, and difference in quality, in agreement with Weber's law. Psychophysical laws, particularly Weber's law, have been found in diverse species, including unicellular organisms. Our theoretical results predict that superorganisms may also exhibit such behaviour, suggesting that these laws arise from fundamental mechanisms of information processing and decision-making. Finally, we propose a combined psychophysical law which unifies Hick-Hyman's law and Piéron's law, traditionally studied independently; this unified law makes predictions that can be empirically tested.

  17. A time-compressed simulated geomagnetic storm influences the nest-exiting flight angles of the stingless bee Tetragonisca angustula

    NASA Astrophysics Data System (ADS)

    Esquivel, D. M. S.; Corrêa, A. A. C.; Vaillant, O. S.; de Melo, V. Bandeira; Gouvêa, G. S.; Ferreira, C. G.; Ferreira, T. A.; Wajnberg, E.

    2014-03-01

    Insects have been used as models for understanding animal orientation. It is well accepted that social insects such as honeybees and ants use different natural cues in their orientation mechanism. A magnetic sensitivity was suggested for the stingless bee Schwarziana quadripunctata, based on the observation of a surprising effect of a geomagnetic storm on the nest-exiting flight angles. Stimulated by this result, in this paper, the effects of a time-compressed simulated geomagnetic storm (TC-SGS) on the nest-exiting flight angles of another stingless bee, Tetragonisca angustula, are presented. Under an applied SGS, either on the horizontal or vertical component of the geomagnetic field, both nest-exiting flight angles, dip and azimuth, are statistically different from those under geomagnetic conditions. The angular dependence of ferromagnetic resonance (FMR) spectra of whole stingless bees shows the presence of organized magnetic nanoparticles in their bodies, which indicates this material as a possible magnetic detector.

  18. A time-compressed simulated geomagnetic storm influences the nest-exiting flight angles of the stingless bee Tetragonisca angustula.

    PubMed

    Esquivel, D M S; Corrêa, A A C; Vaillant, O S; de Melo, V Bandeira; Gouvêa, G S; Ferreira, C G; Ferreira, T A; Wajnberg, E

    2014-03-01

    Insects have been used as models for understanding animal orientation. It is well accepted that social insects such as honeybees and ants use different natural cues in their orientation mechanism. A magnetic sensitivity was suggested for the stingless bee Schwarziana quadripunctata, based on the observation of a surprising effect of a geomagnetic storm on the nest-exiting flight angles. Stimulated by this result, in this paper, the effects of a time-compressed simulated geomagnetic storm (TC-SGS) on the nest-exiting flight angles of another stingless bee, Tetragonisca angustula, are presented. Under an applied SGS, either on the horizontal or vertical component of the geomagnetic field, both nest-exiting flight angles, dip and azimuth, are statistically different from those under geomagnetic conditions. The angular dependence of ferromagnetic resonance (FMR) spectra of whole stingless bees shows the presence of organized magnetic nanoparticles in their bodies, which indicates this material as a possible magnetic detector.

  19. Individual variation in prelaying behaviour and the incidence of floor eggs.

    PubMed

    Cooper, J J; Appleby, M C

    1996-05-01

    1. Floor eggs are a problem in non-cage systems for laying hens, as they require secondary egg collecting. Failure to lay in a well-defined nest site may also be a welfare problem for the hens, but only if their nesting motivation has been thwarted. We investigated the relationships between a hen's prelaying behaviour and its tendency to lay on the floor by recording the behaviour of 20 hens housed individually in wire cages with single littered nest boxes. 3. Most floor eggs (80%) were laid by the same 6 hens. These 6 "floor-layers" performed more nest seeking behaviour, less nest-building behaviour and less sitting prior to oviposition than the 14 hens that consistently laid in nest boxes. 4. The incidence of floor eggs declined with age. Both nest and floor laying hens performed less nest seeking behaviour with age. Floor layers, however, increased their performance of nesting behaviour, whilst nest layers performed less nesting behaviour with age. 5. Floor laying hens behaved as if they found the nest box less attractive than nest-laying hens; perhaps because they had lower nesting motivation, or perhaps because their nesting motivation was as high, but they less readily perceived the nest box as an appropriate nest site.

  20. Large increase in nest size linked to climate change: an indicator of life history, senescence and condition.

    PubMed

    Møller, Anders Pape; Nielsen, Jan Tøttrup

    2015-11-01

    Many animals build extravagant nests that exceed the size required for successful reproduction. Large nests may signal the parenting ability of nest builders suggesting that nests may have a signaling function. In particular, many raptors build very large nests for their body size. We studied nest size in the goshawk Accipiter gentilis, which is a top predator throughout most of the Nearctic. Both males and females build nests, and males provision their females and offspring with food. Nest volume in the goshawk is almost three-fold larger than predicted from their body size. Nest size in the goshawk is highly variable and may reach more than 600 kg for a bird that weighs ca. 1 kg. While 8.5% of nests fell down, smaller nests fell down more often than large nests. There was a hump-shaped relationship between nest volume and female age, with a decline in nest volume late in life, as expected for senescence. Clutch size increased with nest volume. Nest volume increased during 1977-2014 in an accelerating fashion, linked to increasing spring temperature during April, when goshawks build and start reproduction. These findings are consistent with nest size being a reliable signal of parental ability, with large nest size signaling superior parenting ability and senescence, and also indicating climate warming.

  1. Detection of Hepatocyte Clones Containing Integrated Hepatitis B Virus DNA Using Inverse Nested PCR.

    PubMed

    Tu, Thomas; Jilbert, Allison R

    2017-01-01

    Chronic hepatitis B virus (HBV) infection is a major cause of liver cirrhosis and hepatocellular carcinoma (HCC), leading to ~600,000 deaths per year worldwide. Many of the steps that occur during progression from the normal liver to cirrhosis and/or HCC are unknown. Integration of HBV DNA into random sites in the host cell genome occurs as a by-product of the HBV replication cycle and forms a unique junction between virus and cellular DNA. Analyses of integrated HBV DNA have revealed that HCCs are clonal and imply that they develop from the transformation of hepatocytes, the only liver cell known to be infected by HBV. Integrated HBV DNA has also been shown, at least in some tumors, to cause insertional mutagenesis in cancer driver genes, which may facilitate the development of HCC. Studies of HBV DNA integration in the histologically normal liver have provided additional insight into HBV-associated liver disease, suggesting that hepatocytes with a survival or growth advantage undergo high levels of clonal expansion even in the absence of oncogenic transformation. Here we describe inverse nested PCR (invPCR), a highly sensitive method that allows detection, sequencing, and enumeration of virus-cell DNA junctions formed by the integration of HBV DNA. The invPCR protocol is composed of two major steps: inversion of the virus-cell DNA junction and single-molecule nested PCR. The invPCR method is highly specific and inexpensive and can be tailored to DNA extracted from large or small amounts of liver. This procedure also allows detection of genome-wide random integration of any known DNA sequence and is therefore a useful technique for molecular biology, virology, and genetic research.

  2. The Influences of Soil Characteristics on Nest-Site Selection in Painted Turtles (Chrysemys picta)

    NASA Astrophysics Data System (ADS)

    Page, R.

    2017-12-01

    A variety of animals dig nests and lay their eggs in soil, leaving them to incubate and hatch without assistance from the parents. Nesting habitat is important for these organisms many of which exhibit temperature dependent sex determination (TSD) whereby the incubation temperature determines the sex of each hatchling. However, suitable nesting habitat may be limited due to anthropogenic activities and global temperature increases. Soil thermal properties are critical to these organisms and are positively correlated with water retention and soil carbon; carbon-rich soils result in higher incubation temperatures. We investigated nest-site selection in painted turtles (Chrysemys picta) inhabiting an anthropogenic pond in south central Pennsylvania. We surveyed for turtle nests and documented location, depth, width, temperature, canopy coverage, clutch size, and hatch success for a total of 31 turtle nests. To address the influence of soil carbon and particle size on nest selection, we analyzed samples collected from: 1) actual nests that were depredated, 2) false nests, incomplete nests aborted during digging prior to nest completion, and 3) randomized locations. Soil samples were separated into coarse, medium, and fine grain size fractions through a stack of sieves. Samples were combusted in a total carbon analyzer to measure weight percent organic carbon. We found that anthropogenic activity at this site has created homogenous, sandy, compacted soils at the uppermost layer that may limit females' access to appropriate nesting habitat. Turtle nesting activity was limited to a linear region north of the pond and was constrained by an impassable rail line. Relative to other studies, turtle nests were notably shallow (5.8±0.9 cm) and placed close to the pond. Compared to false nests and random locations, turtle-selected sites averaged greater coarse grains (35% compared to 20.24 and 20.57%) and less fine grains (47% compared to 59 and 59, respectively). Despite remarkably high soil carbon along the rail line (47.08%) turtles nested here with slightly higher hatch success. We suggest that the turtles are limited to sandy, compact soils with low heat capacities and may compensate for this by also nesting adjacent to the rail line where high soil carbon could increase incubation temperatures.

  3. Is courtship intensity a signal of male parental care in Red-winged Blackbirds (Agelaius phoeniceus)?

    USGS Publications Warehouse

    Yasukawa, Ken; Knight, Richard L.; Skagen, Susan Knight

    1987-01-01

    We studied Red-winged Blackbird (Agelaius phoeniceus) behavior in marsh and prairie habitats to determine whether courting males provide cues of their nest defense, whether females prefer males that defend nests vigorously, and whether male nest defense affects nest success. We found no evidence that courtship behavior provides cues of male nest defense, or that females prefer males that court intensely. We found evidence that females nesting in marshes prefer males that defend nests intensely, but no evidence that intensely defending males in the prairie habitat achieve high nest success. We found differences, however, between marsh and prairie habitats that suggest a coarse-grained relationship between male parental quality and female choice of mate in this species.

  4. Validation and clinical application of a nested PCR for paracoccidioidomycosis diagnosis in clinical samples from Colombian patients.

    PubMed

    Gaviria, Marcela; Rivera, Vanessa; Muñoz-Cadavid, Cesar; Cano, Luz Elena; Naranjo, Tonny Williams

    2015-01-01

    Paracoccidioidomycosis is a systemic and endemic mycosis, restricted to tropical and subtropical areas of Latin America. The infection is caused by the thermal dimorphic fungus Paracoccidioides brasiliensis and Paracoccidioides lutzii. The diagnosis of paracoccidioidomycosis is usually performed by microscopic examination, culture and immunodiagnostic tests to respiratory specimens, body fluids and/or biopsies; however these methods require laboratory personnel with experience and several days to produce a result. In the present study, we have validated and evaluated a nested PCR assay targeting the gene encoding the Paracoccidioides gp43 membrane protein in 191 clinical samples: 115 samples from patients with proven infections other than paracoccidioidomycosis, 51 samples as negative controls, and 25 samples from patients diagnosed with paracoccidioidomycosis. Additionally, the specificity of the nested PCR assay was also evaluated using purified DNA isolated from cultures of different microorganisms (n=35) previously identified by culture and/or sequencing. The results showed that in our hands, this nested PCR assay for gp43 protein showed specificity and sensitivity rates of 100%. The optimized nested PCR conditions in our laboratory allowed detection down to 1fg of P. brasiliensis DNA. Copyright © 2015 Elsevier Editora Ltda. All rights reserved.

  5. High-tech or field techs: Radio-telemetry is a cost-effective method for reducing bias in songbird nest searching

    USGS Publications Warehouse

    Peterson, Sean M.; Streby, Henry M.; Lehman, Justin A.; Kramer, Gunnar R.; Fish, Alexander C.; Andersen, David E.

    2015-01-01

    We compared the efficacy of standard nest-searching methods with finding nests via radio-tagged birds to assess how search technique influenced our determination of nest-site characteristics and nest success for Golden-winged Warblers (Vermivora chrysoptera). We also evaluated the cost-effectiveness of using radio-tagged birds to find nests. Using standard nest-searching techniques for 3 populations, we found 111 nests in locations with habitat characteristics similar to those described in previous studies: edges between forest and relatively open areas of early successional vegetation or shrubby wetlands, with 43% within 5 m of forest edge. The 83 nests found using telemetry were about half as likely (23%) to be within 5 m of forest edge. We spent little time searching >25 m into forest because published reports state that Golden-winged Warblers do not nest there. However, 14 nests found using telemetry (18%) were >25 m into forest. We modeled nest success using nest-searching method, nest age, and distance to forest edge as explanatory variables. Nest-searching method explained nest success better than nest age alone; we estimated that nests found using telemetry were 10% more likely to fledge young than nests found using standard nest-searching methods. Although radio-telemetry was more expensive than standard nest searching, the cost-effectiveness of both methods differed depending on searcher experience, amount of equipment owned, and bird population density. Our results demonstrate that telemetry can be an effective method for reducing bias in Golden-winged Warbler nest samples, can be cost competitive with standard nest-searching methods in some situations, and is likely to be a useful approach for finding nests of other forest-nesting songbirds.

  6. Island characteristics within wetlands influence waterbird nest success and abundance

    USGS Publications Warehouse

    Hartman, Christopher; Ackerman, Joshua T.; Herzog, Mark

    2016-01-01

    Coastal waterbird populations are threatened by habitat loss and degradation from urban and agricultural development and forecasted sea level rise associated with climate change. Remaining wetlands often must be managed to ensure that waterbird habitat needs, and other ecosystem functions, are met. For many waterbirds, the availability of island nesting habitat is important for conserving breeding populations. We used linear mixed models to investigate the influence of pond and island landscape characteristics on nest abundance and nest success of American avocets (Recurvirostra americana), black-necked stilts (Himantopus mexicanus), and Forster's terns (Sterna forsteri) in San Francisco Bay, California, USA, based on a 9-year dataset that included >9,000 nests. Nest abundance and nest success were greatest within ponds and on individual islands located either <1 km or >4 km from San Francisco Bay. Further, nest abundance was greater within ponds with relatively few islands, and on linear-shaped, highly elongated islands compared to more rounded islands. Nest success was greater on islands located away from the nearest surrounding pond levee. Compared to more rounded islands, linear islands contained more near-water habitat preferred by many nesting waterbirds. Islands located away from pond levees may provide greater protection from terrestrial egg and chick predators. Our results indicate that creating and maintaining a few, relatively small, highly elongated and narrow islands away from mainland levees, in as many wetland ponds as possible would be effective at providing waterbirds with preferred nesting habitat.

  7. Predation risk of artificial ground nests in managed floodplain meadows

    NASA Astrophysics Data System (ADS)

    Arbeiter, Susanne; Franke, Elisabeth

    2018-01-01

    Nest predation highly determines the reproductive success in birds. In agricultural grasslands, vegetation characteristics and management practices influences the predation risk of ground breeders. Little is known so far on the predation pressure on non-passerine nests in tall swards. Investigations on the interaction of land use with nesting site conditions and the habitat selection of nest predators are crucial to develop effective conservation measures for grassland birds. In this study, we used artificial nests baited with quail and plasticine eggs to identify potential predators of ground nests in floodplain meadows and related predation risk to vegetation structure and grassland management. Mean daily predation rate was 0.01 (±0.012) after an exposure duration of 21 days. 70% of all observed nest predations were caused by mammals (Red Fox and mustelids) and 17.5% by avian predators (corvids). Nest sites close to the meadow edge and those providing low forb cover were faced with a higher daily predation risk. Predation risk also increased later in the season. Land use in the preceding year had a significant effect on predation risk, showing higher predation rates on unmanaged sites than on mown sites. Unused meadows probably attract mammalian predators, because they provide a high abundance of small rodents and a more favourable vegetation structure for foraging, increasing also the risk of incidental nest predations. Although mowing operation is a major threat to ground-nesting birds, our results suggest that an annual removal of vegetation may reduce predation risk in the subsequent year.

  8. Effects of nest density, location, and timing on breeding success of Caspian Terns

    USGS Publications Warehouse

    Antolos, Michelle; Roby, D.D.; Lyons, Donald E.; Anderson, Scott K.; Collis, K.

    2006-01-01

    One of the proposed benefits of colonial nesting in birds is the protection afforded against avian predators. This advantage may be counter-balanced by the negative effects of intraspecific aggression on breeding success. Effects of nest density, nest location within the colony, and timing of nest initiation on productivity of Caspian Terns (Sterna caspia) were investigated on Crescent Island in the mid-Columbia River, Washington, USA. In the absence of intense nest predation at the Crescent Island tern colony, it was hypothesized that nest density would be negatively associated with productivity. A rangefinder was used to determine spatial distribution of Caspian Tern nests, and these data used to calculate nest characteristics (nest density, nearest neighbor distance, and distance to colony edge) for a randomly-selected subset of nests monitored for nest chronology and productivity. Productivity did not differ between nests in high- and low-density areas of the colony, and was positively associated with earlier nest initiation. Early nests were more productive, were located in areas of higher nest density, and were further from the colony edge than late nests. The strong effect of timing may have been attributable to seasonal declines in prey resources for terns at this site. Our results suggest that Caspian Terns nesting at the highest densities observed in this study did not incur immediate reproductive costs, despite increased potential for encounters between chicks and aggressive conspecific adults.

  9. Applied Genomics: Data Mining Reveals Species-Specific Malaria Diagnostic Targets More Sensitive than 18S rRNA▿†‡

    PubMed Central

    Demas, Allison; Oberstaller, Jenna; DeBarry, Jeremy; Lucchi, Naomi W.; Srinivasamoorthy, Ganesh; Sumari, Deborah; Kabanywanyi, Abdunoor M.; Villegas, Leopoldo; Escalante, Ananias A.; Kachur, S. Patrick; Barnwell, John W.; Peterson, David S.; Udhayakumar, Venkatachalam; Kissinger, Jessica C.

    2011-01-01

    Accurate and rapid diagnosis of malaria infections is crucial for implementing species-appropriate treatment and saving lives. Molecular diagnostic tools are the most accurate and sensitive method of detecting Plasmodium, differentiating between Plasmodium species, and detecting subclinical infections. Despite available whole-genome sequence data for Plasmodium falciparum and P. vivax, the majority of PCR-based methods still rely on the 18S rRNA gene targets. Historically, this gene has served as the best target for diagnostic assays. However, it is limited in its ability to detect mixed infections in multiplex assay platforms without the use of nested PCR. New diagnostic targets are needed. Ideal targets will be species specific, highly sensitive, and amenable to both single-step and multiplex PCRs. We have mined the genomes of P. falciparum and P. vivax to identify species-specific, repetitive sequences that serve as new PCR targets for the detection of malaria. We show that these targets (Pvr47 and Pfr364) exist in 14 to 41 copies and are more sensitive than 18S rRNA when utilized in a single-step PCR. Parasites are routinely detected at levels of 1 to 10 parasites/μl. The reaction can be multiplexed to detect both species in a single reaction. We have examined 7 P. falciparum strains and 91 P. falciparum clinical isolates from Tanzania and 10 P. vivax strains and 96 P. vivax clinical isolates from Venezuela, and we have verified a sensitivity and specificity of ∼100% for both targets compared with a nested 18S rRNA approach. We show that bioinformatics approaches can be successfully applied to identify novel diagnostic targets and improve molecular methods for pathogen detection. These novel targets provide a powerful alternative molecular diagnostic method for the detection of P. falciparum and P. vivax in conventional or multiplex PCR platforms. PMID:21525225

  10. Excessive nest building is a unique behavioural phenotype in the deer mouse model of obsessive-compulsive disorder.

    PubMed

    Wolmarans, De Wet; Stein, Dan J; Harvey, Brian H

    2016-09-01

    Obsessive-compulsive disorder (OCD) is a phenotypically heterogeneous condition characterised by time-consuming intrusive thoughts and/or compulsions. Irrespective of the symptom type diagnosed, the severity of OCD is characterised by heterogeneity in symptom presentation that complicates diagnosis and treatment. Heterogeneity of symptoms would be invaluable in an animal model. Nest building behaviour forms part of the normal behavioural repertoire of rodents and demonstrates profound between-species differences. However, it has been proposed that within-species differences in nest building behaviour (i.e. aberrant vs. normal nest building) may resemble obsessive-compulsive-like symptoms. In an attempt to investigate whether other obsessive-compulsive-like behaviours are present in an animal model of OCD, or if aberrant nest building behaviour may represent a unique obsessive-compulsive phenotype in such a model, the current study assessed nest building behaviour in high (H, viz obsessive-compulsive) and non (N, viz normal) stereotypical deer mice. Subsequently, 12 N and H animals, respectively, were provided with an excess of cotton wool daily for one week prior to and following four weeks of high-dose oral escitalopram treatment (50 mg/kg/day). Data from the current investigation demonstrate daily nesting activity to be highly variable in deer mice, with stereotypy and nest building being independent behaviours. However, we identified unique aberrant large nest building behaviour in 30% of animals from both cohorts that was attenuated by escitalopram to pre-treatment nesting scores of the larger group. In summary, behavioural and drug-treatment evidence confirms that deer mouse behaviour does indeed resemble symptom heterogeneity related to OCD, and as such expands its face and predictive validity for the disorder. © The Author(s) 2016.

  11. Rapid detection of fungal keratitis with DNA-stabilizing FTA filter paper.

    PubMed

    Menassa, Nardine; Bosshard, Philipp P; Kaufmann, Claude; Grimm, Christian; Auffarth, Gerd U; Thiel, Michael A

    2010-04-01

    Purpose. Polymerase chain reaction (PCR) is increasingly important for the rapid detection of fungal keratitis. However, techniques of specimen collection and DNA extraction before PCR may interfere with test sensitivity. The purpose of this study was to investigate the use of DNA-stabilizing FTA filter paper (Indicating FTA filter paper; Whatman International, Ltd., Maidstone, UK) for specimen collection without DNA extraction in a single-step, nonnested PCR for fungal keratitis. Methods. Specimens were collected from ocular surfaces with FTA filter discs, which automatically lyse collected cells and stabilize nucleic acids. Filter discs were directly used in single-step PCR reactions to detect fungal DNA. Test sensitivity was evaluated with serial dilutions of Candida albicans, Fusarium oxysporum, and Aspergillus fumigatus cultures. Test specificity was analyzed by comparing 196 and 155 healthy individuals from Switzerland and Egypt, respectively, with 15 patients with a diagnosis of microbial keratitis. Results. PCR with filter discs detected 3 C. albicans, 25 F. oxysporum, and 125 A. fumigatus organisms. In healthy volunteers, fungal PCR was positive in 1.0% and 8.4% of eyes from Switzerland and Egypt, respectively. Fungal PCR remained negative in 10 cases of culture-proven bacterial keratitis, became positive in 4 cases of fungal keratitis, but missed 1 case of culture-proven A. fumigatus keratitis. Conclusions. FTA filter paper for specimen collection together with direct PCR is a promising method of detecting fungal keratitis. The analytical sensitivity is high without the need for a semi-nested or nested second PCR, the clinical specificity is 91.7% to 99.0%, and the method is rapid and inexpensive.

  12. Methods for Integrated Air Sampling and DNA Analysis for Detection of Airborne Fungal Spores

    PubMed Central

    Williams, Roger H.; Ward, Elaine; McCartney, H. Alastair

    2001-01-01

    Integrated air sampling and PCR-based methods for detecting airborne fungal spores, using Penicillium roqueforti as a model fungus, are described. P. roqueforti spores were collected directly into Eppendorf tubes using a miniature cyclone-type air sampler. They were then suspended in 0.1% Nonidet P-40, and counted using microscopy. Serial dilutions of the spores were made. Three methods were used to produce DNA for PCR tests: adding untreated spores to PCRs, disrupting spores (fracturing of spore walls to release the contents) using Ballotini beads, and disrupting spores followed by DNA purification. Three P. roqueforti-specific assays were tested: single-step PCR, nested PCR, and PCR followed by Southern blotting and probing. Disrupting the spores was found to be essential for achieving maximum sensitivity of the assay. Adding untreated spores to the PCR did allow the detection of P. roqueforti, but this was never achieved when fewer than 1,000 spores were added to the PCR. By disrupting the spores, with or without subsequent DNA purification, it was possible to detect DNA from a single spore. When known quantities of P. roqueforti spores were added to air samples consisting of high concentrations of unidentified fungal spores, pollen, and dust, detection sensitivity was reduced. P. roqueforti DNA could not be detected using untreated or disrupted spore suspensions added to the PCRs. However, using purified DNA, it was possible to detect 10 P. roqueforti spores in a background of 4,500 other spores. For all DNA extraction methods, nested PCR was more sensitive than single-step PCR or PCR followed by Southern blotting. PMID:11375150

  13. Nest Predation Deviates from Nest Predator Abundance in an Ecologically Trapped Bird.

    PubMed

    Hollander, Franck A; Van Dyck, Hans; San Martin, Gilles; Titeux, Nicolas

    2015-01-01

    In human-modified environments, ecological traps may result from a preference for low-quality habitat where survival or reproductive success is lower than in high-quality habitat. It has often been shown that low reproductive success for birds in preferred habitat types was due to higher nest predator abundance. However, between-habitat differences in nest predation may only weakly correlate with differences in nest predator abundance. An ecological trap is at work in a farmland bird (Lanius collurio) that recently expanded its breeding habitat into open areas in plantation forests. This passerine bird shows a strong preference for forest habitat, but it has a higher nest success in farmland. We tested whether higher abundance of nest predators in the preferred habitat or, alternatively, a decoupling of nest predator abundance and nest predation explained this observed pattern of maladaptive habitat selection. More than 90% of brood failures were attributed to nest predation. Nest predator abundance was more than 50% higher in farmland, but nest predation was 17% higher in forest. Differences between nest predation on actual shrike nests and on artificial nests suggested that parent shrikes may facilitate nest disclosure for predators in forest more than they do in farmland. The level of caution by parent shrikes when visiting their nest during a simulated nest predator intrusion was the same in the two habitats, but nest concealment was considerably lower in forest, which contributes to explaining the higher nest predation in this habitat. We conclude that a decoupling of nest predator abundance and nest predation may create ecological traps in human-modified environments.

  14. Nest Predation Deviates from Nest Predator Abundance in an Ecologically Trapped Bird

    PubMed Central

    Hollander, Franck A.; Van Dyck, Hans; San Martin, Gilles; Titeux, Nicolas

    2015-01-01

    In human-modified environments, ecological traps may result from a preference for low-quality habitat where survival or reproductive success is lower than in high-quality habitat. It has often been shown that low reproductive success for birds in preferred habitat types was due to higher nest predator abundance. However, between-habitat differences in nest predation may only weakly correlate with differences in nest predator abundance. An ecological trap is at work in a farmland bird (Lanius collurio) that recently expanded its breeding habitat into open areas in plantation forests. This passerine bird shows a strong preference for forest habitat, but it has a higher nest success in farmland. We tested whether higher abundance of nest predators in the preferred habitat or, alternatively, a decoupling of nest predator abundance and nest predation explained this observed pattern of maladaptive habitat selection. More than 90% of brood failures were attributed to nest predation. Nest predator abundance was more than 50% higher in farmland, but nest predation was 17% higher in forest. Differences between nest predation on actual shrike nests and on artificial nests suggested that parent shrikes may facilitate nest disclosure for predators in forest more than they do in farmland. The level of caution by parent shrikes when visiting their nest during a simulated nest predator intrusion was the same in the two habitats, but nest concealment was considerably lower in forest, which contributes to explaining the higher nest predation in this habitat. We conclude that a decoupling of nest predator abundance and nest predation may create ecological traps in human-modified environments. PMID:26624619

  15. Edge, height and visibility effects on nest predation by birds and mammals in the Brazilian cerrado

    NASA Astrophysics Data System (ADS)

    Dodonov, Pavel; Paneczko, Ingrid Toledo; Telles, Marina

    2017-08-01

    Edge influence is one of the main impacts in fragmented landscapes; yet, most of studies on edge influence have focused on high-contrast edges, and the impacts of low-contrast edges and narrow linear openings are less understood. Edge influence often affects bird nest predation, but these effects are not ubiquitous and may depend on characteristics such as nest height and visibility. We performed an experiment on nest predation in a migratory passerine, Elaenia chiriquensis (Lesser Elaenia; Passeriformes: Tyrannidae), in a savanna vegetation of the Brazilian Cerrado biome in South-Eastern Brazil. We used 89 real E. chiriquensis nests, collected during previous reproductive seasons, with two plasticine eggs in each, and randomly distributed them at two locations (edge - up to 20 m from a firebreak edge and interior - approx. 150-350 m from the edge) and two heights (low - 60-175 cm and high - 190-315 cm above ground). We also measured leaf and branch density around each nest. We performed this study on two 15-days campaigns, checking the nests every 2-3 days and removing those with predation marks. We sorted the predation marks into those made by birds, mammals, or unidentified predators, and used generalized linear models to assess the effects of location, height and leaf density on survival time and predator type. Only four nests had not been predated during the experiment; 55 nests were predated by birds, 7 by mammals, and 23 by unidentified predators. Low nests in the interior tended to have larger survival times whereas high nests at the edge tended to be more predated by birds and less predated by mammals. Thus, even a low-contrast (firebreak) edge may significantly increase nest predation, which is also affected by the nest's height, mainly due to predation by birds. These effects may be due to predator movement along the edge as well as to edge-related changes in vegetation structure. We suggest that higher-contrast edges which may also be used as movement conduits, for example powerline openings, may have even stronger effects, demanding further studies.

  16. Comparison of milk culture, direct and nested polymerase chain reaction (PCR) with fecal culture based on samples from dairy herds infected with Mycobacterium avium subsp. paratuberculosis

    PubMed Central

    Gao, Anli; Odumeru, Joseph; Raymond, Melinda; Hendrick, Steven; Duffield, Todd; Mutharia, Lucy

    2009-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne’s disease in cattle and other farm ruminants, and is also a suspected pathogen of Crohn’s disease in humans. Development of diagnostic methods for MAP infection has been a challenge over the last few decades. The objective of this study was to investigate the relationship between different methods for detection of MAP in milk and fecal samples. A total of 134 milk samples and 110 feces samples were collected from 146 individual cows in 14 MAP-infected herds in southwestern Ontario. Culture, IS900 polymerase chain reaction (PCR) and nested PCR methods were used for detecting MAP in milk; results were compared with those of fecal culture. A significant relationship was found between milk culture, direct PCR, and nested PCR (P < 0.05). The fecal culture results were not related to any of the 3 assay methods used for the milk samples (P > 0.10). Although fecal culture showed a higher sensitivity than the milk culture method, the difference was not significant (P = 0.2473). The number of MAP colony-forming units (CFU) isolated by culture from fecal samples was, on average, higher than that isolated from milk samples (P = 0.0083). There was no significant correlation between the number of CFU cultured from milk and from feces (Pearson correlation coefficient = 0.1957, N = 63, P = 0.1243). The animals with high numbers of CFU in milk culture may not be detected by fecal culture at all, and vise versa. A significant proportion (29% to 41%) of the positive animals would be missed if only 1 culture method, instead of both milk and feces, were to be used for diagnosis. This suggests that the shedding of MAP in feces and milk is not synchronized. Most of the infected cows were low-level shedders. The proportion of low-level shedders may even be underestimated because MAP is killed during decontamination, thus reducing the chance of detection. Therefore, to identify suspected Johne’s-infected animals using the tests in this study, both milk and feces samples should be collected in duplicate to enhance the diagnostic rate. The high MAP kill rate identified in the culture methods during decontamination may be compensated for by using the nested PCR method, which had a higher sensitivity than the IS900 PCR method used. PMID:19337397

  17. Detection by hemi-nested reverse transcription polymerase chain reaction and genetic characterization of wild type strains of Canine distemper virus in suspected infected dogs.

    PubMed

    Di Francesco, Cristina E; Di Francesco, Daniela; Di Martino, Barbara; Speranza, Roberto; Santori, Domenico; Boari, Andrea; Marsilio, Fulvio

    2012-01-01

    A new highly sensitive and specific hemi-nested reverse transcription polymerase chain reaction (RT-PCR) assay was applied to detect nucleoprotein (NP) gene of Canine distemper virus (CDV) in samples collected from dogs showing respiratory, gastrointestinal, and neurological signs. Thirty-eight out of 86 samples were positive suggesting that despite the vaccination, canine distemper may still represent a high risk to the canine population. The 968 base pair (bp) fragments from the hemagglutinin (H) gene of 10 viral strains detected in positive samples were amplified and analyzed by restriction fragment length polymorphism (RFLP) using AluI and PsiI enzymes in order to differentiate among vaccine and wild-type CDV strains and to characterize the field viral strains. The products of the both enzymatic digestions allowed identification all viruses as wild strains of CDV. In addition, the RFLP analysis with AluI provided additional information about the identity level among the strains analyzed on the basis of the positions of the cleavage site in the nucleotide sequences of the H gene. The method could be a more useful and simpler method for molecular studies of CDV strains.

  18. Detection and molecular characterization of Cryptosporidium spp. in captive canaries (Serinus canaria) using different diagnostic methods.

    PubMed

    Camargo, Vinícius da Silva; Santana, Bruna Nicoleti; Ferrari, Elis Domingos; Nakamura, Alex Akira; Nagata, Walter Bertequini; Nardi, Ana Rita Moraes; Meireles, Marcelo Vasconcelos

    2018-01-01

    This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.

  19. Nested archetypes of vulnerability in African drylands: where lies potential for sustainable agricultural intensification?

    NASA Astrophysics Data System (ADS)

    Sietz, D.; Ordoñez, J. C.; Kok, M. T. J.; Janssen, P.; Hilderink, H. B. M.; Tittonell, P.; Van Dijk, H.

    2017-09-01

    Food production is key to achieving food security in the drylands of sub-Saharan Africa. Since agricultural productivity is limited, however, due to inherent agro-ecological constraints and land degradation, sustainable agricultural intensification has been widely discussed as an opportunity for improving food security and reducing vulnerability. Yet vulnerability determinants are distributed heterogeneously in the drylands of sub-Saharan Africa and sustainable intensification cannot be achieved everywhere in cost-effective and efficient ways. To better understand the heterogeneity of farming systems’ vulnerability in order to support decision making at regional scales, we present archetypes, i.e. socio-ecological patterns, of farming systems’ vulnerability in the drylands of sub-Saharan Africa and reveal their nestedness. We quantitatively indicated the most relevant farming systems’ properties at a sub-national resolution. These factors included water availability, agro-ecological potential, erosion sensitivity, population pressure, urbanisation, remoteness, governance, income and undernourishment. Cluster analysis revealed eight broad archetypes of vulnerability across all drylands of sub-Saharan Africa. The broad archetype representing better governance and highest remoteness in extremely dry and resource-constrained regions encompassed the largest area share (19%), mainly indicated in western Africa. Moreover, six nested archetypes were identified within those regions with better agropotential and prevalent agricultural livelihoods. Among these patterns, the nested archetype depicting regions with highest erosion sensitivity, severe undernourishment and lower agropotential represented the largest population (30%) and area (28%) share, mainly found in the Sahel region. The nested archetype indicating medium undernourishment, better governance and lowest erosion sensitivity showed particular potential for sustainable agricultural intensification, mainly in western and some parts of southeastern and eastern Africa. Insights into the nestedness of archetypes allowed a more differentiated discussion of vulnerability and sustainable intensification opportunities, enhancing the evaluation of key interlinkages between land management and food security. The archetypes may support the transfer of successful intensification strategies based on similarities among the drylands in sub-Saharan Africa.

  20. Comparison of the diagnostic performance of microscopic examination with nested polymerase chain reaction for optimum malaria diagnosis in Upper Myanmar.

    PubMed

    Kang, Jung-Mi; Cho, Pyo-Yun; Moe, Mya; Lee, Jinyoung; Jun, Hojong; Lee, Hyeong-Woo; Ahn, Seong Kyu; Kim, Tae Im; Pak, Jhang Ho; Myint, Moe Kyaw; Lin, Khin; Kim, Tong-Soo; Na, Byoung-Kuk

    2017-03-16

    Accurate diagnosis of Plasmodium infection is crucial for prompt malaria treatment and surveillance. Microscopic examination has been widely applied as the gold standard for malaria diagnosis in most part of malaria endemic areas, but its diagnostic value has been questioned, particularly in submicroscopic malaria. In this study, the diagnostic performance of microscopic examination and nested polymerase chain reaction (PCR) was evaluated to establish optimal malaria diagnosis method in Myanmar. A total of 1125 blood samples collected from residents in the villages and towns located in Naung Cho, Pyin Oo Lwin, Tha Beik Kyin townships and Mandalay of Upper Myanmar were screened by microscopic examination and species-specific nested PCR method. Among the 1125 blood samples, 261 samples were confirmed to be infected with malaria by microscopic examination. Evaluation of the 1125 samples by species-specific nested PCR analysis revealed that the agreement between microscopic examination and nested PCR was 87.3% (261/299). Nested PCR successfully detected 38 Plasmodium falciparum or Plasmodium vivax infections, which were missed in microscopic examination. Microscopic examinations also either misdiagnosed the infected Plasmodium species, or did not detect mixed infections with different Plasmodium species in 31 cases. The nested PCR method is more reliable than conventional microscopic examination for the diagnosis of malaria infections, and this is particularly true in cases of mixed infections and submicroscopic infections. Given the observed higher sensitivity and specificity of nested PCR, the molecular method holds enormous promise in malaria diagnosis and species differentiation, and can be applied as an effective monitoring tool for malaria surveillance, control and elimination in Myanmar.

  1. An economical wireless cavity-nest viewer

    Treesearch

    Daniel P. Huebner; Sarah R. Hurteau

    2007-01-01

    Inspection of cavity nests and nest boxes is often required during studies of cavity-nesting birds, and fiberscopes and pole-mounted video cameras are sometimes used for such inspection. However, the cost of these systems may be prohibitive for some potential users. We describe a user-built, wireless cavity viewer that can be used to access cavities as high as 15 m and...

  2. Nesting behavior of house mice (Mus domesticus) selected for increased wheel-running activity.

    PubMed

    Carter, P A; Swallow, J G; Davis, S J; Garland, T

    2000-03-01

    Nest building was measured in "active" (housed with access to running wheels) and "sedentary" (without wheel access) mice (Mus domesticus) from four replicate lines selected for 10 generations for high voluntary wheel-running behavior, and from four randombred control lines. Based on previous studies of mice bidirectionally selected for thermoregulatory nest building, it was hypothesized that nest building would show a negative correlated response to selection on wheel-running. Such a response could constrain the evolution of high voluntary activity because nesting has also been shown to be positively genetically correlated with successful production of weaned pups. With wheel access, selected mice of both sexes built significantly smaller nests than did control mice. Without wheel access, selected females also built significantly smaller nests than did control females, but only when body mass was excluded from the statistical model, suggesting that body mass mediated this correlated response to selection. Total distance run and mean running speed on wheels was significantly higher in selected mice than in controls, but no differences in amount of time spent running were measured, indicating a complex cause of the response of nesting to selection for voluntary wheel running.

  3. Changes in infestation, cell cap condition, and reproductive status of Varroa destructor (Mesostigmata: Varrroidae) in brood exposed to honey bees with Varroa sensitive hygiene

    USDA-ARS?s Scientific Manuscript database

    Honey bees (Apis mellifera L.) bred for Varroa sensitive hygiene (VSH) selectively remove pupae infested with Varroa destructor Anderson & Trueman from capped brood that is inserted into the nest. After one week, remaining brood cells tend to have been uncapped and recapped, and remaining mites are...

  4. The effect of canopy closure on chimpanzee nest abundance in Lagoas de Cufada National Park, Guinea-Bissau.

    PubMed

    Sousa, Joana; Casanova, Catarina; Barata, André V; Sousa, Cláudia

    2014-04-01

    The present study aimed to gather baseline information about chimpanzee nesting and density in Lagoas de Cufada Natural Park (LCNP), in Guinea-Bissau. Old and narrow trails were followed to estimate chimpanzee density through marked-nest counts and to test the effect of canopy closure (woodland savannah, forest with a sparse canopy, and forest with a dense canopy) on nest distribution. Chimpanzee abundance was estimated at 0.79 nest builders/km(2), the lowest among the areas of Guinea-Bissau with currently studied chimpanzee populations. Our data suggest that sub-humid forest with a dense canopy accounts for significantly higher chimpanzee nest abundance (1.50 nests/km of trail) than sub-humid forest with a sparse canopy (0.49 nests/km of trail) or woodland savannah (0.30 nests/km of trail). Dense-canopy forests play an important role in chimpanzee nesting in the patchy and highly humanized landscape of LCNP. The tree species most frequently used for nesting are Dialium guineense (46%) and Elaeis guineensis (28%). E. guineensis contain nests built higher in the canopy, while D. guineense contain nests built at lower heights. Nests observed during baseline sampling and replications suggest seasonal variations in the tree species used for nest building.

  5. Performance of two rapid diagnostic tests for malaria diagnosis at the China-Myanmar border area

    PubMed Central

    2013-01-01

    Background Rapid diagnostic tests (RDTs) have become an essential tool in the contemporary malaria control and management programmes in the world. This study aims to evaluate the performance of two commonly used RDTs for malaria diagnosis in the China-Myanmar border area. Methods A total 606 febrile patients in the China-Myanmar border were recruited to this study and were diagnosed for malaria infections by microscopy, two RDTs tests (Pf/Pan device, and Pv/Pf device) and nested PCR. Results Malaria parasites were found in 143 patients by microscopy, of which 51, 73, and 19 were Plasmodium falciparum, Plasmodium vivax and P. falciparum/P. vivax mixed infections, respectively. Compared to microscopy, the sensitivity of the Pf/Pan device was 88.6% for P. falciparum and 69.9% for P. vivax with the specificity of 90.4%. For a subset of 350 patients, the sensitivity of the Pf/Pan device and Pv/Pf device for detection of P. falciparum was 87.5% and 91.7%, respectively; and for detection of P. vivax was 72.0% and 73.8%, respectively. The specificity of the Pf/Pan device and Pv/Pf device was 94.3% and 96.5%, respectively. Nested PCR detected malaria parasites in 174 of 606 samples, of which 67, 79, two and 26 were P. falciparum, P. vivax, P. ovale and P. falciparum/P. vivax mixed infections, respectively. Compared to nested PCR, all other methods had sensitivity below 80%, suggesting that a significant number of cases were missed. Conclusions Compared to PCR, both microscopy and RDTs had lower sensitivities. RDTs had similar performance to microscopy for P. falciparum diagnosis, but performed worse for P. vivax diagnosis. Other RDT products should be selected with higher sensitivity (and good specificity) for both P. falciparum and P. vivax diagnosis. PMID:23433230

  6. The role of moisture in the nest thermoregulation of social wasps.

    PubMed

    Klingner, R; Richter, K; Schmolz, E; Keller, B

    2005-09-01

    Paper nests of social wasps are intriguing constructions for both, biologists and engineers. We demonstrate that moisture and latent heat significantly influence the thermal performance of the nest construction. Two colonies of the hornet Vespa crabro were investigated in order to clarify the relation of the temperature and the moisture regime inside the nest. Next to fairly stable nest temperatures the hornets maintain a high relative humidity inside the nest. We found that in consequence a partial vapor-pressure gradient between nest and ambient drives a constant vapor flux through the envelope. The vapor flux is limited by the diffusion resistance of the envelope. The driving force of vapor flux is heat, which is consumed through evaporation inside the nest. The colony has to compensate this loss with metabolic heat production in order to maintain a stable nest temperature. However, humidity fluctuations inside the nest induce circadian adsorption and desorption cycles, which stabilize the nest temperature and thus contribute significantly to temperature homeostasis. Our study demonstrates that both mechanisms influence nest thermoregulation and need to be considered to understand the thermodynamic behavior of nests of wasps and social insects in general.

  7. Strategies for nest-site selection by king eiders

    USGS Publications Warehouse

    Bentzen, R.L.; Powell, A.N.; Suydam, R.S.

    2009-01-01

    Nest site selection is a critical component of reproduction and has presumably evolved in relation to predation, local resources, and microclimate. We investigated nest-site choice by king eiders (Somateria spectabilis) on the coastal plain of northern Alaska, USA, 2003-2005. We hypothesized that nest-site selection is driven by predator avoidance and that a variety of strategies including concealment, seclusion, and conspecific or inter-specific nest defense might lead to improved nesting success. We systematically searched wetland basins for king eider nests and measured habitat and social variables at nests (n = 212) and random locations (n = 493). King eiders made use of both secluded and concealed breeding strategies; logistic regression models revealed that females selected nests close to water, on islands, and in areas with high willow (Salix spp.) cover but did not select sites near conspecific or glaucous gull (Larus hyperboreus) nests. The most effective nest-placement strategy may vary depending on density and types of nest predators; seclusion is likely a mammalian-predator avoidance tactic whereas concealment may provide protection from avian predators. We recommend that managers in northern Alaska attempt to maintain wetland basins with islands and complex shorelines to provide potential nest sites in the vicinity of water. ?? The Wildlife Society.

  8. Effects of industrial and investigator disturbance on Arctic-nesting geese

    USGS Publications Warehouse

    Meixell, Brandt W.; Flint, Paul L.

    2017-01-01

    Oil and gas development on the Arctic Coastal Plain (ACP) of Alaska, USA may have effects on Arctic-nesting birds. To estimate effects of industrial activity and investigator disturbance on avian productivity, we monitored nests of greater white-fronted geese (Anser albifrons) with digital cameras and periodic nest visits during 2013–2014 at 2 sites on the ACP. A disturbed site was adjacent to human-made infrastructure and industrial clean-up activities initiated at the onset of the study and a control site was >2 km from sources of industrial disturbance. We assessed variation in estimates of incubation constancy, nest survival, and predator behavior relative to site, year, and distance from industrial activity using nest photographs obtained at 1-minute intervals. We compared analysis of hourly nest survival informed by intensive monitoring with cameras to analysis of daily nest survival informed by traditional nest visit data obtained at intervals of 5–7 days to assess how method and time scale of sampling affect ecological inference. Geese in both sites exhibited high levels of nest attendance and initiated incubation breaks less than once per day. Observer-caused incubation breaks associated with nest visits ( = 37.8 min) were longer than other types of incubation breaks ( = 8.7 min), demonstrating a differential response by nesting geese to direct human encroachment versus indirect vehicular and aircraft traffic. During both years, geese were absent from nests more frequently in the disturbed ( = 0.9 breaks/day) than control ( = 0.6 breaks/day) site, and this break frequency was slightly higher for nests closer to industrial activity. In the year with high rates of depredation, nest survival was positively related to distance from industrial activity and abandoned infrastructure, consistent with predictions of industry-caused effects. This relationship, however, was not evident in the year with reduced predation pressure, likely because of annual variation in arctic fox (Vulpes lagopus) behavior. Analysis of nest survival probability informed by camera data allowed for detection of detailed patterns of variation that were not supported when using only visit data for the same nests. Observer visits were responsible for reductions of 7–35% in nest survival probability, highlighting the importance of minimizing, and controlling for, observer effects in studies of avian productivity. Indirect vehicular and aircraft disturbance posed less risk to nest survival than direct encroachment by observers at nest sites. Therefore, effects of industrial activities on avian productivity in the Arctic can be minimized through practices that limit direct encounters with nests. 

  9. Nesting ecology of Chelonia mydas (Testudines: Cheloniidae) on the Guanahacabibes Peninsula, Cuba.

    PubMed

    Azanza Ricardo, Julia; Ibarra Martín, Maria E; González Sansón, Gaspar; Abreu Grobois, F Alberto; Eckert, Karen L; Espinosa López, Georgina; Oyama, Ken

    2013-12-01

    The nesting colony of green sea turtles (Chelonia mydas) at Guanahacabibes Peninsula Biosphere Reserve and National Park is one of the largest in the Cuban archipelago; however, little information about its nesting ecology is available. Temporal and spatial variation in nesting and reproductive success as well as morphometric characteristics of gravid females were used to ecologically characterize this colony. Nine beaches of the Southernmost coast of Guanahacabibes Peninsula were monitored for 14 years (1998-2012) to determine green turtle nesting activity, from May to September (peak nesting season in this area). Beach dimensions were measured to determine nest density using the length and the area. Afterward the beaches were divided in two categories, index and secondary. Females were measured and tagged to compare new tagged females (823) with returning tagged females (140). Remigration interval was also determined. Temporal variation was identified as the annual number of nesting emergences and oviposits per female, with apparent peaks in reproductive activity on a biennial cycle in the first six years followed by periods of annual increase in nest number (2003-2008) and periods of decreasing number of nests (2010-2012). We also found intra-seasonal variation with the highest nesting activity in July, particularly in the second half of the month. The peak emergence time was 22:00-02:00 hr. In terms of spatial variation, smaller beaches had the highest nest density and nesting was more frequent 6-9m from the high tide line, where hatchling production was maximized although hatchling success was high on average, above 80%. Morphometric analysis of females was made and newly tagged turtles were smaller on average than remigrants. Our results are only a first attempt at characterizing Guanahacabibes' populations but have great value for establishing conservation priorities within the context of national management plans, and for efficient monitoring and protection of nesting beaches.

  10. Marine turtles are not fussy nesters: a novel test of small-scale nest site selection using structure from motion beach terrain information

    PubMed Central

    Leon, Javier X.; Gilby, Ben L.; Olds, Andrew D.; Schlacher, Thomas A.

    2017-01-01

    Background Nest selection is widely regarded as a key process determining the fitness of individuals and viability of animal populations. For marine turtles that nest on beaches, this is particularly pivotal as the nesting environment can significantly control reproductive success.The aim of this study was to identify the environmental attributes of beaches (i.e., morphology, vegetation, urbanisation) that may be associated with successful oviposition in green and loggerhead turtle nests. Methods We quantified the proximity of turtle nests (and surrounding beach locations) to urban areas, measured their exposure to artificial light, and used ultra-high resolution (cm-scale) digital surface models derived from Structure-from-Motion (SfM) algorithms, to characterise geomorphic and vegetation features of beaches on the Sunshine Coast, eastern Australia. Results At small spatial scales (i.e., <100 m), we found no evidence that turtles selected nest sites based on a particular suite of environmental attributes (i.e., the attributes of nest sites were not consistently different from those of surrounding beach locations). Nest sites were, however, typically characterised by occurring close to vegetation, on parts of the shore where the beach- and dune-face was concave and not highly rugged, and in areas with moderate exposure to artificial light. Conclusion This study used a novel empirical approach to identify the attributes of turtle nest sites from a broader ‘envelope’ of environmental nest traits, and is the first step towards optimizing conservation actions to mitigate, at the local scale, present and emerging human impacts on turtle nesting beaches. PMID:28070454

  11. Industrious leaf cutter ants and their carbon footprints

    NASA Astrophysics Data System (ADS)

    Swanson, A. C.; Dierick, D.; Trahan, N. A.; Allen, M. F.; Schwendenmann, L.; Harmon, T. C.; Oberbauer, S. F.; Fernandez Bou, A. S.; Zelikova, T. J.

    2017-12-01

    Leaf cutter ants (LCA) are considered ecosystem engineers in Neotropical forest ecosystems because they alter physical and environmental conditions for other organisms. LCA excavate large underground nests, maintaining intricate tunnels and fungal and waste chambers, and they continuously bring in vast amounts of fresh leaf material. In order to understand their ecosystem-wide impacts, we set out to determine whether their engineering activities fundamentally alter soil structure, soil nutrient pools, and gas fluxes in a wet tropical rainforest in Costa Rica. To directly compare LCA nest to non-nest sites, we utilized embedded sensor arrays with series of soil moisture, CO2, O2, and temperature sensors placed at four soil depths and automated minirhizotrons (AMR) to measure root and hyphal production and turnover. We also collected soils for biogeochemical analyses and measured soil CO2 fluxes and carbon isotope ratios of below-ground CO2 for two years. Our measurements confirmed that LCA alter their soil environment to regulate internal soil CO2 concentrations, moisture, and temperature, increasing O2 concentrations in the process. There were marked differences in soil structure inside nests relative to non-nests and these were associated with increased root and hyphal production and turnover in nests. Soil C, N, P, and their respective degrading enzymes were highly variable among sites and between nests and controls but N and P increased with soil depth and were generally higher in nests than controls. Contrary to our expectations, C mineralization rates were lower in nests but CO2 fluxes were high from nest vents and similar to non-nests elsewhere. At the system scale, LCA appear to fundamentally change the soil environment inside their nests and create spatial heterogeneity in biogeochemical processes and root and hyphal growth, influencing the overall C balance of Neotropical forests.

  12. Predation and infanticide influence ideal free choice by a parrot occupying heterogeneous tropical habitats.

    PubMed

    Bonebrake, Timothy C; Beissinger, Steven R

    2010-06-01

    The ideal free distribution (IFD) predicts that organisms will disperse to sites that maximize their fitness based on availability of resources. Habitat heterogeneity underlies resource variation and influences spatial variation in demography and the distribution of populations. We relate nest site productivity at multiple scales measured over a decade to habitat quality in a box-nesting population of Forpus passerinus (green-rumped parrotlets) in Venezuela to examine critical IFD assumptions. Variation in reproductive success at the local population and neighborhood scales had a much larger influence on productivity (fledglings per nest box per year) than nest site or female identity. Habitat features were reliable cues of nest site quality. Nest sites with less vegetative cover produced greater numbers of fledglings than sites with more cover. However, there was also a competitive cost to nesting in high-quality, low-vegetative cover nest boxes, as these sites experienced the most infanticide events. In the lowland local population, water depth and cover surrounding nest sites were related with F. passerinus productivity. Low vegetative cover and deeper water were associated with lower predation rates, suggesting that predation could be a primary factor driving habitat selection patterns. Parrotlets also demonstrated directional dispersal. Pairs that changed nest sites were more likely to disperse from poor-quality nest sites to high-quality nest sites rather than vice versa, and juveniles were more likely to disperse to, or remain in, the more productive of the two local populations. Parrotlets exhibited three characteristics fundamental to the IFD: habitat heterogeneity within and between local populations, reliable habitat cues to productivity, and active dispersal to sites of higher fitness.

  13. Compound Eye Adaptations for Diurnal and Nocturnal Lifestyle in the Intertidal Ant, Polyrhachis sokolova

    PubMed Central

    Narendra, Ajay; Alkaladi, Ali; Raderschall, Chloé A.; Robson, Simon K. A.; Ribi, Willi A.

    2013-01-01

    The Australian intertidal ant, Polyrhachis sokolova lives in mudflat habitats and nests at the base of mangroves. They are solitary foraging ants that rely on visual cues. The ants are active during low tides at both day and night and thus experience a wide range of light intensities. We here ask the extent to which the compound eyes of P. sokolova reflect the fact that they operate during both day and night. The ants have typical apposition compound eyes with 596 ommatidia per eye and an interommatidial angle of 6.0°. We find the ants have developed large lenses (33 µm in diameter) and wide rhabdoms (5 µm in diameter) to make their eyes highly sensitive to low light conditions. To be active at bright light conditions, the ants have developed an extreme pupillary mechanism during which the primary pigment cells constrict the crystalline cone to form a narrow tract of 0.5 µm wide and 16 µm long. This pupillary mechanism protects the photoreceptors from bright light, making the eyes less sensitive during the day. The dorsal rim area of their compound eye has specialised photoreceptors that could aid in detecting the orientation of the pattern of polarised skylight, which would assist the animals to determine compass directions required while navigating between nest and food sources. PMID:24155883

  14. Comparison of diagnostic techniques for the detection of Cryptosporidium oocysts in animal samples

    PubMed Central

    Mirhashemi, Marzieh Ezzaty; Zintl, Annetta; Grant, Tim; Lucy, Frances E.; Mulcahy, Grace; De Waal, Theo

    2015-01-01

    While a large number of laboratory methods for the detection of Cryptosporidium oocysts in faecal samples are now available, their efficacy for identifying asymptomatic cases of cryptosporidiosis is poorly understood. This study was carried out to determine a reliable screening test for epidemiological studies in livestock. In addition, three molecular tests were compared to identify Cryptosporidium species responsible for the infection in cattle, sheep and horses. A variety of diagnostic tests including microscopic (Kinyoun's staining), immunological (Direct Fluorescence Antibody tests or DFAT), enzyme-linked immunosorbent assay (ELISA), and molecular methods (nested PCR) were compared to assess their ability to detect Cryptosporidium in cattle, horse and sheep faecal samples. The results indicate that the sensitivity and specificity of each test is highly dependent on the input samples; while Kinyoun's and DFAT proved to be reliable screening tools for cattle samples, DFAT and PCR analysis (targeted at the 18S rRNA gene fragment) were more sensitive for screening sheep and horse samples. Finally different PCR primer sets targeted at the same region resulted in the preferential amplification of certain Cryptosporidium species when multiple species were present in the sample. Therefore, for identification of Cryptosporidium spp. in the event of asymptomatic cryptosporidiosis, the combination of different 18S rRNA nested PCR primer sets is recommended for further epidemiological applications and also tracking the sources of infection. PMID:25662435

  15. Beach dynamics and nest distribution of the leatherback turtle (Dermochelys coriacea) at Grande Riviere Beach, Trinidad & Tobago.

    PubMed

    Lum, Lori Lee

    2005-05-01

    Grande Riviere Beach in Trinidad and Tobago is an important nesting site in the Caribbean for the Critically Endangered leatherback sea turtle, Dermochelys coriacea. Community members were concerned that beach erosion and seasonal river flooding were destroying many of the nests deposited annually and thought that a hatchery was a possible solution. Over the 2001 turtle nesting season, the Institute of Marine Affairs (IMA) assessed the spatial and temporal distribution of nests using the Global Positioning System recorded to reference points, and beach dynamics using permanent bench mark profile stations, to determine areas of high risk and more stable areas for nesting. A total of 1449 leatherback nests were positioned. It was evident that at the start of the season in March, the majority of leatherback nests were deposited at the eastern section of the beach. After May, there was a continuing westward shift in nest distribution as the season progressed until August and beach erosion in the eastern section became predominant. The backshore remained relatively stable along the entire beach throughout the nesting season, and erosion was predominant in the foreshore at the eastern section of the beach, from the middle to the end of the season. Similar trends in accretion and erosion were observed in 2000. River flooding did not occur during the study period or in the previous year. With both high risk and more stable regions for turtle nesting available at Grande Riviere Beach, there was no compelling evidence to justify the need for a hatchery.

  16. Testing models of female reproductive migratory behaviour and population structure in the Caribbean hawksbill turtle, Eretmochelys imbricata, with mtDNA sequences.

    PubMed

    Bass, A L; Good, D A; Bjorndal, K A; Richardson, J I; Hillis, Z M; Horrocks, J A; Bowen, B W

    1996-06-01

    Information on the reproductive behaviour and population structure of female hawksbill turtles, Eretmochelys imbricata, is necessary to define conservation priorities for this highly endangered species. Two hypotheses to explain female nest site choice, natal homing and social facilitation, were tested by analyzing mtDNA control region sequences of 103 individuals from seven nesting colonies in the Caribbean and western Atlantic. Under the social facilitation model, newly mature females follow older females to a nesting location, and subsequently use this site for future nesting. This model generates an expectation that female lineages will be homogenized among regional nesting colonies. Contrary to expectations of the social facilitation model, mtDNA lineages were highly structured among western Atlantic nesting colonies. These analyses identified at least 6 female breeding stocks in the Caribbean and western Atlantic and support a natal homing model for recruitment of breeding females. Reproductive populations are effectively isolated over ecological time scales, and recovery plans for this species should include protection at the level of individual nesting colonies.

  17. Anadromous sea lampreys (Petromyzon marinus) are ecosystem engineers in a spawning tributary

    USGS Publications Warehouse

    Hogg, Robert S.; Coghlan, Stephen M.; Zydlewski, Joseph D.; Simon, Kevin S.

    2014-01-01

    Sea lampreys (Petromyzon marinus) disturb the substratum during nest construction and alter the physical habitat, potentially affecting other stream organisms. We quantified differences in depth, velocity, fine-sediment coverage, embeddedness, intragravel permeability and benthic invertebrate assemblages (density and diversity) among nest mounds, nest pits and undisturbed reference locations over a 4-month period after June spawning. In 2010 and 2011, immediate and persistent effects of nest construction were assessed in summer (July) and in autumn (late September to early October), respectively. Randomly selected nests were sampled annually (25 each in summer and autumn). Nest construction increased stream-bed complexity by creating and juxtaposing shallow, swift, rocky habitat patches with deep, slow, sandy habitat patches. Mounds had a 50–143% less cover of fine sediment, and a 30–62% reduction in embeddedness, compared to pits and reference locations. These physical changes persisted into the autumn (almost 4 months). Five insect families contributed 74% of the benthic invertebrate abundance: Chironomidae (27%), Hydropsychidae (26%), Heptageniidae (8%), Philopotamidae (7%) and Ephemerellidae (6%). Densities of Hydropsychidae, Philopotamidae and Heptageniidae were up to 10 times greater in mounds than in pits and adjacent reference habitat. In summer, mounds had twice the density of Chironomidae than did pits, and 1.5 times more than reference habitats, but densities were similar among the habitats in autumn. These results suggest that spawning sea lampreys are ecosystem engineers. The physical disturbance caused by nest-building activity was significant and persistent, increasing habitat heterogeneity and favouring pollution-sensitive benthic invertebrates and, possibly, drift-feeding fish.

  18. Nest substrate reflects incubation style in extant archosaurs with implications for dinosaur nesting habits.

    PubMed

    Tanaka, Kohei; Zelenitsky, Darla K; Therrien, François; Kobayashi, Yoshitsugu

    2018-03-15

    Dinosaurs thrived and reproduced in various regions worldwide, including the Arctic. In order to understand their nesting in diverse or extreme environments, the relationships between nests, nesting environments, and incubation methods in extant archosaurs were investigated. Statistical analyses reveal that species of extant covered nesters (i.e., crocodylians and megapodes) preferentially select specific sediments/substrates as a function of their nesting style and incubation heat sources. Relationships between dinosaur eggs and the sediments in which they occur reveal that hadrosaurs and some sauropods (i.e., megaloolithid eggs) built organic-rich mound nests that relied on microbial decay for incubation, whereas other sauropods (i.e., faveoloolithid eggs) built sandy in-filled hole nests that relied on solar or potentially geothermal heat for incubation. Paleogeographic distribution of mound nests and sandy in-filled hole nests in dinosaurs reveals these nest types produced sufficient incubation heat to be successful up to mid latitudes (≤47°), 10° higher than covered nesters today. However, only mound nesting and likely brooding could have produced sufficient incubation heat for nesting above the polar circle (>66°). As a result, differences in nesting styles may have placed restrictions on the reproduction of dinosaurs and their dispersal at high latitudes.

  19. High atmospheric temperatures and ‘ambient incubation’ drive embryonic development and lead to earlier hatching in a passerine bird

    PubMed Central

    Griffith, Simon C.; Mainwaring, Mark C.; Sorato, Enrico; Beckmann, Christa

    2016-01-01

    Tropical and subtropical species typically experience relatively high atmospheric temperatures during reproduction, and are subject to climate-related challenges that are largely unexplored, relative to more extensive work conducted in temperate regions. We studied the effects of high atmospheric and nest temperatures during reproduction in the zebra finch. We characterized the temperature within nests in a subtropical population of this species in relation to atmospheric temperature. Temperatures within nests frequently exceeded the level at which embryo’s develop optimally, even in the absence of parental incubation. We experimentally manipulated internal nest temperature to demonstrate that an average difference of 6°C in the nest temperature during the laying period reduced hatching time by an average of 3% of the total incubation time, owing to ‘ambient incubation’. Given the avian constraint of laying a single egg per day, the first eggs of a clutch are subject to prolonged effects of nest temperature relative to later laid eggs, potentially increasing hatching asynchrony. While birds may ameliorate the negative effects of ambient incubation on embryonic development by varying the location and design of their nests, high atmospheric temperatures are likely to constitute an important selective force on avian reproductive behaviour and physiology in subtropical and tropical regions, particularly in the light of predicted climate change that in many areas is leading to a higher frequency of hot days during the periods when birds breed. PMID:26998315

  20. Seasonal fecundity is not related to geographic position ...

    EPA Pesticide Factsheets

    AimSixty-five years ago, Theodosius Dobzhansky suggested that individuals of a species face greater challenges from abiotic stressors at high latitudes and from biotic stressors at their low-latitude range edges. This idea has been expanded to the hypothesis that species’ ranges are limited by abiotic and biotic stressors at high and low latitudes, respectively. Support has been found in many systems, but this hypothesis has almost never been tested with demographic data. We present an analysis of fecundity across the breeding range of a species as a test of this hypothesis.Location575 km of tidal marshes in the northeastern United States.MethodsWe monitored saltmarsh sparrow (Ammodramus caudacutus) nests at twenty-three sites from Maine to New Jersey, USA. With data from 840 nests, we calculated daily nest failure probabilities due to competing abiotic (flooding) and biotic (depredation) stressors.ResultsWe observed that abiotic stress (nest flooding probability) was greater than biotic stress (nest depredation probability) at the high-latitude range edge of saltmarsh sparrows, consistent with Dobzhansky’s hypothesis. Similarly, biotic stress decreased with increasing latitude throughout the range, whereas abiotic stress was not predicted by latitude alone. Instead, nest flooding probability was best predicted by date, maximum high tide, and extremity of rare flooding events.Main conclusionsOur results provide support for Dobzhansky’s hypothesis across th

  1. Comparative evaluation of uniplex, nested, semi-nested, multiplex and nested multiplex PCR methods in the identification of microbial etiology of clinically suspected infectious endophthalmitis.

    PubMed

    Bharathi, Madasamy Jayahar; Murugan, Nandagopal; Rameshkumar, Gunasekaran; Ramakrishnan, Rengappa; Venugopal Reddy, Yerahaia Chinna; Shivkumar, Chandrasekar; Ramesh, Srinivasan

    2013-05-01

    This study is aimed to determine the utility of various polymerase chain reaction (PCR) methods in vitreous fluids (VFs) for detecting the infectious genomes in the diagnosis of infectious endophthalmitis in terms of sensitivity and specificity. This prospective and consecutive analysis included a total of 66 VFs that were submitted for the microbiological evaluation, which were obtained from 66 clinically diagnosed endophthalmitis patients presented between November 2010 and October 2011 at the tertiary eye care referral centre in South India. Part of the collected VFs were subjected to cultures and smears, and the remaining parts were utilized for five PCR methods: uniplex, nested, semi-nested, multiplex and nested multiplex after extracting DNA, using universal eubacterial and Propionibacterium acnes species-specific primer sets targeting 16S rRNA gene in all bacteria and P. acnes, and panfungal primers, targeting 28S rRNA gene in all fungi. Of the 66 VFs, five (7.5%) showed positive results in smears, 16 (24%) in cultures and 43 (65%) showed positive results in PCRs. Among the 43 positively amplified VFs, 10 (15%) were positive for P. acnes genome, one for panfungal genome and 42 (62%) for eubacterial genome (including 10 P. acnes positives). Among 42 eubacterial-positive VFs, 36 were positive by both uniplex (first round) and multiplex (first round) PCRs, while nested (second round) and nested multiplex (second round) PCRs produced positive results in 42 and 41 VFs, respectively. Of the 43 PCR-positive specimens, 16 (37%) had positive growth (15 bacterial and one fungal) in culture. Of 50 culture-negative specimens, 27 (54%) were showed positive amplification, of which 10 were amplified for both P. acnes and eubacterial genomes and the remaining 17 were for eubacterial genome alone. Nested PCRs are superior than uniplex and multiplex PCR. PCRs proved to be a powerful tool in the diagnosis of endophthalmitis, especially for detecting uncultured microbes.

  2. Phenology of marine turtle nesting revealed by statistical model of the nesting season.

    PubMed

    Girondot, Marc; Rivalan, Philippe; Wongsopawiro, Ronald; Briane, Jean-Paul; Hulin, Vincent; Caut, Stéphane; Guirlet, Elodie; Godfrey, Matthew H

    2006-08-31

    Marine turtles deposit their eggs on tropical or subtropical beaches during discrete nesting seasons that span several months. The number and distribution of nests laid during a nesting season provide vital information on various aspects of marine turtle ecology and conservation. In the case of leatherback sea turtles nesting in French Guiana, we developed a mathematical model to explore the phenology of their nesting season, derived from an incomplete nest count dataset. We detected 3 primary components in the nest distribution of leatherbacks: an overall shape that corresponds to the arrival and departure of leatherback females in the Guianas region, a sinusoidal pattern with a period of approximately 10 days that is related to physiological constraints of nesting female leatherbacks, and a sinusoidal pattern with a period of approximately 15 days that likely reflects the influence of spring high tides on nesting female turtles. The model proposed here offers a variety of uses for both marine turtles and also other taxa when individuals are observed in a particular location for only part of the year.

  3. Sensitive detection of Treponema pallidum DNA from the whole blood of patients with syphilis by the nested PCR assay.

    PubMed

    Wang, Cuini; Cheng, Yuanyuan; Liu, Biao; Wang, Yuanyuan; Gong, Weiming; Qian, Yihong; Guan, Zhifang; Lu, Haikong; Gu, Xin; Shi, Mei; Zhou, Pingyu

    2018-05-09

    The aim of this work was to investigate the application of the nested PCR assay for the detection of Treponema pallidum (TP) DNA from the blood of patients with different stages of syphilis. In this study, a nested PCR method targeting the Tpp47 and polA genes (Tpp47-Tp-PCR and polA-Tp-PCR) was developed to detect TP-DNA in whole blood samples collected from 262 patients with different stages of syphilis (84 primary syphilis, 97 secondary syphilis, and 81 latent syphilis patients). The PCR assay detected T. pallidum DNA in 53.6% and 62.9% of the patients with primary and secondary syphilis, respectively, which was much higher than the detection levels in patients with latent syphilis (7.4%) (both p < 0.001). For primary syphilis, a low RPR (0-16) was correlated with a higher detection rate of TP-DNA, whereas for secondary syphilis, the higher detection rate of blood TP-DNA was correlated with higher blood RPR titers (at or beyond 32). For latent syphilis, TP-DNA was only detectable by PCR in the early phase of the latent infection. Thus, blood RPR titers were correlated with the blood T. pallidum burden, but the correlations varied with primary and secondary syphilis. The results indicate that nested PCR is a sensitive method for detecting blood TP-DNA and is especially useful for detecting early syphilis including primary syphilis and secondary syphilis. The findings also suggest that the PCR assay may be used to complement other methods to enhance the diagnosis of syphilis.

  4. Presence of Breeding Birds Improves Body Condition for a Crocodilian Nest Protector.

    PubMed

    Nell, Lucas A; Frederick, Peter C; Mazzotti, Frank J; Vliet, Kent A; Brandt, Laura A

    2016-01-01

    Ecological associations where one species enhances habitat for another nearby species (facilitations) shape fundamental community dynamics and can promote niche expansion, thereby influencing how and where species persist and coexist. For the many breeding birds facing high nest-predation pressure, enemy-free space can be gained by nesting near more formidable animals for physical protection. While the benefits to protected species seem well documented, very few studies have explored whether and how protector species are affected by nest protection associations. Long-legged wading birds (Pelecaniformes and Ciconiiformes) actively choose nesting sites above resident American alligators (Alligator mississippiensis), apparently to take advantage of the protection from mammalian nest predators that alligator presence offers. Previous research has shown that wading bird nesting colonies could provide substantial food for alligators in the form of dropped chicks. We compared alligator body condition in similar habitat with and without wading bird nesting colonies present. Alligator morphometric body condition indices were significantly higher in colony than in non-colony locations, an effect that was statistically independent of a range of environmental variables. Since colonially nesting birds and crocodilians co-occur in many tropical and subtropical wetlands, our results highlight a potentially widespread keystone process between two ecologically important species-groups. These findings suggest the interaction is highly beneficial for both groups of actors, and illustrate how selective pressures may have acted to form and reinforce a strongly positive ecological interaction.

  5. Projecting avian response to linked changes in groundwater and riparian floodplain vegetation along a dryland river: A scenario analysis

    USGS Publications Warehouse

    Arriana, Brand L.; Stromberg, J.C.; Goodrich, D.C.; Dixon, M.D.; Lansey, K.; Kang, D.; Brookshire, D.S.; Cerasale, D.J.

    2011-01-01

    Groundwater is a key driver of riparian condition on dryland rivers but is in high demand for municipal, industrial, and agricultural uses. Approaches are needed to guide decisions that balance human water needs while conserving riparian ecosystems. We developed a space-for-time substitution model that links groundwater change scenarios implemented within a Decision Support System (DSS) with proportions of floodplain vegetation types and abundances of breeding and migratory birds along the upper San Pedro River, AZ, USA. We investigated nine scenarios ranging from groundwater depletion to recharge. In groundwater decline scenarios, relative proportions of tall-canopied obligate phreatophytes (Populus/Salix, cottonwood/willow) on the floodplain progressively decline, and shrubbier species less dependent on permanent water sources (e.g. Tamarix spp., saltcedar) increase. These scenarios result in broad shifts in the composition of the breeding bird community, with canopy-nesting and water-obligate birds declining but midstory nesting birds increasing in abundance as groundwater declines. For the most extreme draw-down scenario where all reaches undergo groundwater declines, models project that only 10% of the upper San Pedro floodplain would be comprised of cottonwood/willow (73% saltcedar and 18% mesquite), and abundances of canopy-nesting, water-obligate, and spring migrant birds would decline 48%, 72%, and 40%, respectively. Groundwater recharge scenarios were associated with increases in canopy-nesting birds particularly given the extreme recharge scenario (all reaches regain shallow water tables and perennial streamflow). Model outputs serve to assess the sensitivity of biotic groups to potential changes in groundwater and thus to rank scenarios based on their expected ecological impacts. ?? 2010 John Wiley & Sons, Ltd.

  6. Nests and nest sites of the San Miguel Island Song Sparrow

    USGS Publications Warehouse

    Kern, Michael D.; Sogge, Mark K.; Kern, Robert B.; van Riper, Charles

    1993-01-01

    Nests and nest sites of the San Miguel Island (SMI) Song Sparrow (Melospiza melodia micronyx) are described; nests are compared with those of 16 other races of Song Sparrows. Bush lupins (Lupinus albifrons), coyote brush (Baccharis pilularis) and golden bush (Haplopappus venetus) were the shrubs used most commonly as nest sites by Song Sparrows on SMI. As a result of its location, the nest was effectively concealed from gray foxes (Urocyon littoralis), the major predator of this sparrow. Nest and nest site also moderated the combined chilling effects of cool air temperatures and strong northwesterly winds on the eggs and nestlings. Even in the absence of these moderating effects of the nest site, the energetic cost of incubation, estimated at 41-53% of the sparrow's resting metabolic rate, was modest. Twenty-nine percent of the canopy above the nest was open and as much as 73% of the nest cup was in the sun at midday, a time when surface temperatures of foliage, nest and nestlings sometimes exceeded 40 C. Whereas this exposure did not apparently reduce fledging success, it may explain why the incidence of addled eggs was so high in this population of Song Sparrows compared to others. Significant differences existed among races of Song Sparrows in the size, porosity and insulation of the nest. In most cases, these differences were not related to the latitude of the races' nesting areas.

  7. Variability in nest survival rates and implications to nesting studies

    USGS Publications Warehouse

    Klett, A.T.; Johnson, D.H.

    1982-01-01

    We used four reasonably large samples (83-213) of Mallard (Anas platyrhynchos) and Blue-winged Teal (A. discors) nests on an interstate highway right-of-way in southcentral North Dakota to evaluate potential biases in hatch-rate estimates. Twelve consecutive, weekly searches for nests were conducted with a cable-chain drag in 1976 and 1977. Nests were revisited at weekly intervals. Four methods were used to estimate hatch rates for the four data sets: the Traditional Method, the Mayfield Method, and two modifications of the Mayfield Method that are sometimes appropriate when daily mortality rates of nests are not constant. Hatch rates and the average age of nests at discovery declined as the interval between searches decreased, suggesting that mortality rates were not constant in our samples. An analysis of variance indicated that daily mortality rates varied with the age of nests in all four samples. Mortality was generally highest during the early laying period, moderately high during the late laying period, and lowest during incubation. We speculate that this relationship of mortality to nest age might be due to the presence of hens at nests or to differences in the vulnerability of nest sites to predation. A modification of the Mayfield Method that accounts for age-related variation in nest mortality was most appropriate for our samples. We suggest methods for conducting nesting studies and estimating nest success for species possessing similar nesting habits.

  8. Predation and nutrients drive population declines in breeding waders.

    PubMed

    Møller, Anders Pape; Thorup, Ole; Laursen, Karsten

    2018-04-20

    Allee effects are defined as a decline in per capita fitness at low population density. We hypothesized that predation reduces population size of breeding waders and thereby the efficiency of predator deterrence, while total nitrogen through its effects on primary and secondary productivity increases population size. Therefore, nest predation could have negative consequences for population size because nest failure generally results in breeding dispersal and hence reduced local population density. To test these predictions, we recorded nest predation in five species of waders for 4,745 nests during 1987-2015 at the nature reserve Tipperne, Denmark. Predation rates were generally negatively related to conspecific and heterospecific population density, but positively related to overall population density of the entire wader community. Nest predation and population density were related to ground water level, management (grazing and mowing), and nutrients. High nest predation with a time lag of one year resulted in low overall breeding population density, while high nutrient levels resulted in higher population density. These two factors accounted for 86% of the variance in population size, presumably due to effects of nest predation on emigration, while nutrient levels increased the level of vegetation cover and the abundance of food in the surrounding brackish water. These findings are consistent with the hypothesis that predation may reduce population density through negative density dependence, while total nitrogen at adjacent shallow water may increase population size. Nest predation rates were reduced by high ground water level in March, grazing by cattle and mowing that affected access to and susceptibility of nests to predators. These effects can be managed to benefit breeding waders. © 2018 by the Ecological Society of America.

  9. Living with aliens: effects of invasive shrub honeysuckles on avian nesting.

    PubMed

    Gleditsch, Jason M; Carlo, Tomás A

    2014-01-01

    Invasive species have come to the forefront of conservation biology as a major threat to native biodiversity. Habitats dominated by shrub honeysuckles (Lonicera spp.) in the United States have been characterized as "ecological traps" by ecologists. Here we tested this hypothesis by investigating the effects of shrub honeysuckles on the nesting ecology of native birds in seven study sites in central Pennsylvania, USA. We examined how the abundance of shrub honeysuckles influenced the selection of nesting substrates and habitat for a community of common songbirds, and the parental-care behavior and nestling development of gray catbirds (Dumetella carolinensis). We found that birds had a strong bias towards nesting in honeysuckle shrubs, but not necessarily for nesting in honeysuckle-dominated habitats. Nest predation rates were affected by the density of nests in a habitat, but not by the overall abundance of honeysuckles in such habitats. Honeysuckle abundance in the habitat did show significant effects on some parental-care behavioral parameters: catbirds had higher nest visitation rates and shorter visit lengths in areas of high honeysuckle density. On average, Gray catbirds fed fruit 12%±0.31 s.e. of their nestling-feeding bouts, mostly fruits of shrub honeysuckles. Nestlings in sites with high honeysuckle density also showed higher mass:tarsus ratios, suggesting a good (possibly better) physiological condition of catbird nestlings at the time of fledging. Our study shows that honeysuckle-dominated habitats could have equivocal effects on nesting parameters of common species of native birds. We advise more caution in the widespread denomination of novel plant communities with high densities of honeysuckle as "ecological traps" as effects can be null or positive on native birds in certain localities.

  10. Nest site attributes and temporal patterns of northern flicker nest loss: effects of predation and competition.

    PubMed

    Fisher, Ryan J; Wiebe, Karen L

    2006-04-01

    To date, most studies of nest site selection have failed to take into account more than one source of nest loss (or have combined all sources in one analysis) when examining nest site characteristics, leaving us with an incomplete understanding of the potential trade-offs that individuals may face when selecting a nest site. Our objectives were to determine whether northern flickers (Colaptes auratus) may experience a trade-off in nest site selection in response to mammalian nest predation and nest loss to a cavity nest competitor (European starling, Sturnus vulgaris). We also document within-season temporal patterns of these two sources of nest loss with the hypothesis that flickers may also be constrained in the timing of reproduction under both predatory and competitive influence. Mammalian predators frequently depredated flicker nests that were: lower to the ground, less concealed by vegetation around the cavity entrance and at the base of the nest tree, closer to coniferous forest edges and in forest clumps with a high percentage of conifer content. Proximity to coniferous edges or coniferous trees increased the probability of nest predation, but nests near conifers were less likely to be lost to starlings. Flickers may thus face a trade-off in nest site selection with respect to safety from predators or competitors. Models suggested that peaks of nest predation and nest loss to eviction occurred at the same time, although a competing model suggested that the peak of nest loss to starlings occurred 5 days earlier than the peak of mammalian predation. Differences in peaks of mammalian predation and loss to starlings may constrain any adjustment in clutch initiation date by flickers to avoid one source of nest loss.

  11. A New Diagnostic system for Ultra Sensitive and Specific Detection and Quantitation of “Candidatus Liberibacter asiaticus”, the Bacterium Associated with Citrus Huanglongbing

    USDA-ARS?s Scientific Manuscript database

    In this study, an ultra sensitive and quantitative diagnostic system for “Candidatus Liberibacter asiaticus” was developed. This system adapts a nested PCR and Taq-Man PCR in a single closed tube. The procedure involves two steps of PCR using the species specific outer and inner primer pairs. Differ...

  12. Does body size influence nest attendance? A comparison of Ross's geese (Chen rossii) and the larger, sympatric lesser snow geese (C. caerulescens caerulescens)

    USGS Publications Warehouse

    Jonsson, J.E.; Afton, A.D.; Alisauskas, R.T.

    2007-01-01

    The body-size hypothesis predicts that nest attendance is positively related to body size among waterfowl and that recess duration is inversely related to body size. Several physiological and behavioral characteristics of Ross's geese (Chen rossii) suggest that females of this species should maintain high nest attendance despite their relatively small body size. Accordingly, we used 8-mm films to compare the incubation behavior of Ross's geese to that of the larger, closely-related lesser snow geese (C. caerulescens caerulescens; hereafter, snow geese) nesting sympatrically at Karrak lake, Nunavut, Canada in 1993. We found that nest attendance averaged 99% for both species. Our results offer no support for the body-size hypothesis. We suggest that temperature requirements of embryos in relation to short incubation duration and a low foraging efficiency of females select for high nest attendance in both snow geese and Ross's geese. ?? Dt. Ornithologen-Gesellschaft e.V. 2007.

  13. Changes in reproductive life-history strategies in response to nest density in a shell-brooding cichlid, Telmatochromis vittatus

    NASA Astrophysics Data System (ADS)

    Ota, Kazutaka; Hori, Michio; Kohda, Masanori

    2012-01-01

    To determine whether the appearance of a reproductively parasitic tactic varies, and how this variation affects territorial males of the Lake Tanganyika cichlid fish Telmatochromis vittatus, we examined the reproductive ecology of territorial males in Mtondwe and compared it with that of a neighboring Wonzye population, where nest density differs from that at Mtondwe. In Wonzye, with high nest density, male tactics change with their body size from a territorial to a non-territorial parasitic tactic called piracy in which they conquer several nests defended by territorial males and take over the nests while females are spawning. These "pirate" males could decrease the costs incurred by travelling among nests by exclusively targeting aggregations of nests in close proximity while avoiding separate nests. Territorial males in Wonzye sacrifice the potential higher attractiveness offered by large nests and instead compete for nests farther from neighbors on which pirates less frequently intrude. In contrast, the Mtondwe population had lower nest density and piracy was absent. Given that the success of piracy depends on the close proximity of nests, nest density is likely responsible for the observed variation in the occurrence of piracy between the two populations. Furthermore, in Mtondwe, territorial males competed for larger nests and were smaller than the territorial males in Wonzye. Thus, this lower nest density may free territorial males from the selection pressures for increased size caused by both defense against nest piracy and the need to develop into pirates as they grow.

  14. Simulation and Sensitivity in a Nested Modeling System for South America. Part II: GCM Boundary Forcing.

    NASA Astrophysics Data System (ADS)

    Rojas, Maisa; Seth, Anji

    2003-08-01

    of this study, the RegCM's ability to simulate circulation and rainfall observed in the two extreme seasons was demonstrated when driven at the lateral boundaries by reanalyzed forcing. Seasonal integrations with the RegCM driven by GCM ensemble-derived lateral boundary forcing demonstrate that the nested model responds well to the SST forcing, by capturing the major features of the circulation and rainfall differences between the two years. The GCM-driven model also improves upon the monthly evolution of rainfall compared with that from the GCM. However, the nested model rainfall simulations for the two seasons are degraded compared with those from the reanalyses-driven RegCM integrations. The poor location of the Atlantic intertropical convergence zone (ITCZ) in the GCM leads to excess rainfall in Nordeste in the nested model.An expanded domain was tested, wherein the RegCM was permitted more internal freedom to respond to SST and regional orographic forcing. Results show that the RegCM is able to improve the location of the ITCZ, and the seasonal evolution of rainfall in Nordeste, the Amazon region, and the southeastern region of Brazil. However, it remains that the limiting factor in the skill of the nested modeling system is the quality of the lateral boundary forcing provided by the global model.

  15. Detection of fungi by conventional methods and semi-nested PCR in patients with presumed fungal keratitis.

    PubMed

    Haghani, I; Amirinia, F; Nowroozpoor-Dailami, K; Shokohi, T

    2015-06-01

    Fungal keratitis is a suppurative, ulcerative, and sight-threatening infection of the cornea that sometimes leads to blindness. The aims of this study were: recuperating facilities for laboratory diagnosis, determining the causative microorganisms, and comparing conventional laboratory diagnostic tools and semi-nested PCR. Sampling was conducted in patients with suspected fungal keratitis. Two corneal scrapings specimens, one for direct smear and culture and the other for semi- nested PCR were obtained. Of the 40 expected cases of mycotic keratitis, calcofluor white staining showed positivity in 25%, culture in 17.5%, KOH in 10%, and semi-nested PCR in 27.5%. The sensitivities of semi-nested PCR, KOH, and CFW were 57.1%, 28.5%, and 42% while the specificities were 78.7%, 94%, and 78.7%, respectively. The time taken for PCR assay was 4 to 8 hours, whereas positive fungal cultures took at least 5 to 7 days. Due to the increasing incidence of fungal infections in people with weakened immune systems, uninformed using of topical corticosteroids and improper use of contact lens, fast diagnosis and accurate treatment of keratomycosis seems to be essential . Therefore, according to the current study, molecular methods can detect mycotic keratitis early and correctly leading to appropriate treatment.

  16. Improved assay to detect Plasmodium falciparum using an uninterrupted, semi-nested PCR and quantitative lateral flow analysis

    PubMed Central

    2013-01-01

    Background A rapid, non-invasive, and inexpensive point-of-care (POC) diagnostic for malaria followed by therapeutic intervention would improve the ability to control infection in endemic areas. Methods A semi-nested PCR amplification protocol is described for quantitative detection of Plasmodium falciparum and is compared to a traditional nested PCR. The approach uses primers that target the P. falciparum dihydrofolate reductase gene. Results This study demonstrates that it is possible to perform an uninterrupted, asymmetric, semi-nested PCR assay with reduced assay time to detect P. falciparum without compromising the sensitivity and specificity of the assay using saliva as a testing matrix. Conclusions The development of this PCR allows nucleic acid amplification without the need to transfer amplicon from the first PCR step to a second reaction tube with nested primers, thus reducing both the chance of contamination and the time for analysis to < two hours. Analysis of the PCR amplicon yield was adapted to lateral flow detection using the quantitative up-converting phosphor (UCP) reporter technology. This approach provides a basis for migration of the assay to a POC microfluidic format. In addition the assay was successfully evaluated with oral samples. Oral fluid collection provides a simple non-invasive method to collect clinical samples. PMID:23433252

  17. Influence of olfactory and visual cover on nest site selection and nest success for grassland-nesting birds.

    PubMed

    Fogarty, Dillon T; Elmore, R Dwayne; Fuhlendorf, Samuel D; Loss, Scott R

    2017-08-01

    Habitat selection by animals is influenced by and mitigates the effects of predation and environmental extremes. For birds, nest site selection is crucial to offspring production because nests are exposed to extreme weather and predation pressure. Predators that forage using olfaction often dominate nest predator communities; therefore, factors that influence olfactory detection (e.g., airflow and weather variables, including turbulence and moisture) should influence nest site selection and survival. However, few studies have assessed the importance of olfactory cover for habitat selection and survival. We assessed whether ground-nesting birds select nest sites based on visual and/or olfactory cover. Additionally, we assessed the importance of visual cover and airflow and weather variables associated with olfactory cover in influencing nest survival. In managed grasslands in Oklahoma, USA, we monitored nests of Northern Bobwhite ( Colinus virginianus ), Eastern Meadowlark ( Sturnella magna ), and Grasshopper Sparrow ( Ammodramus savannarum ) during 2015 and 2016. To assess nest site selection, we compared cover variables between nests and random points. To assess factors influencing nest survival, we used visual cover and olfactory-related measurements (i.e., airflow and weather variables) to model daily nest survival. For nest site selection, nest sites had greater overhead visual cover than random points, but no other significant differences were found. Weather variables hypothesized to influence olfactory detection, specifically precipitation and relative humidity, were the best predictors of and were positively related to daily nest survival. Selection for overhead cover likely contributed to mitigation of thermal extremes and possibly reduced detectability of nests. For daily nest survival, we hypothesize that major nest predators focused on prey other than the monitored species' nests during high moisture conditions, thus increasing nest survival on these days. Our study highlights how mechanistic approaches to studying cover informs which dimensions are perceived and selected by animals and which dimensions confer fitness-related benefits.

  18. Will the Effects of Sea-Level Rise Create Ecological Traps for Pacific Island Seabirds?

    PubMed

    Reynolds, Michelle H; Courtot, Karen N; Berkowitz, Paul; Storlazzi, Curt D; Moore, Janet; Flint, Elizabeth

    2015-01-01

    More than 18 million seabirds nest on 58 Pacific islands protected within vast U.S. Marine National Monuments (1.9 million km2). However, most of these seabird colonies are on low-elevation islands and sea-level rise (SLR) and accompanying high-water perturbations are predicted to escalate with climate change. To understand how SLR may impact protected islands and insular biodiversity, we modeled inundation and wave-driven flooding of a globally important seabird rookery in the subtropical Pacific. We acquired new high-resolution Digital Elevation Models (DEMs) and used the Delft3D wave model and ArcGIS to model wave heights and inundation for a range of SLR scenarios (+0.5, +1.0, +1.5, and +2.0 m) at Midway Atoll. Next, we classified vegetation to delineate habitat exposure to inundation and identified how breeding phenology, colony synchrony, and life history traits affect species-specific sensitivity. We identified 3 of 13 species as highly vulnerable to SLR in the Hawaiian Islands and quantified their atoll-wide distribution (Laysan albatross, Phoebastria immutabilis; black-footed albatross, P. nigripes; and Bonin petrel, Pterodroma hypoleuca). Our models of wave-driven flooding forecast nest losses up to 10% greater than passive inundation models at +1.0 m SLR. At projections of + 2.0 m SLR, approximately 60% of albatross and 44% of Bonin petrel nests were overwashed displacing more than 616,400 breeding albatrosses and petrels. Habitat loss due to passive SLR may decrease the carrying capacity of some islands to support seabird colonies, while sudden high-water events directly reduce survival and reproduction. This is the first study to simulate wave-driven flooding and the combined impacts of SLR, groundwater rise, and storm waves on seabird colonies. Our results highlight the need for early climate change planning and restoration of higher elevation seabird refugia to prevent low-lying protected islands from becoming ecological traps in the face of rising sea levels.

  19. Will the effects of sea-level rise create ecological traps for Pacific Island seabirds?

    USGS Publications Warehouse

    Reynolds, Michelle H.; Courtot, Karen; Berkowitz, Paul; Storlazzi, Curt; Moore, Janet; Flint, Elizabeth

    2015-01-01

    More than 18 million seabirds nest on 58 Pacific islands protected within vast U.S. Marine National Monuments (1.9 million km2). However, most of these seabird colonies are on low-elevation islands and sea-level rise (SLR) and accompanying high-water perturbations are predicted to escalate with climate change. To understand how SLR may impact protected islands and insular biodiversity, we modeled inundation and wave-driven flooding of a globally important seabird rookery in the subtropical Pacific. We acquired new high-resolution Digital Elevation Models (DEMs) and used the Delft3D wave model and ArcGIS to model wave heights and inundation for a range of SLR scenarios (+0.5, +1.0, +1.5, and +2.0 m) at Midway Atoll. Next, we classified vegetation to delineate habitat exposure to inundation and identified how breeding phenology, colony synchrony, and life history traits affect species-specific sensitivity. We identified 3 of 13 species as highly vulnerable to SLR in the Hawaiian Islands and quantified their atoll-wide distribution (Laysan albatross, Phoebastria immutabilis; black-footed albatross, P. nigripes; and Bonin petrel, Pterodroma hypoleuca). Our models of wave-driven flooding forecast nest losses up to 10% greater than passive inundation models at +1.0 m SLR. At projections of + 2.0 m SLR, approximately 60% of albatross and 44% of Bonin petrel nests were overwashed displacing more than 616,400 breeding albatrosses and petrels. Habitat loss due to passive SLR may decrease the carrying capacity of some islands to support seabird colonies, while sudden high-water events directly reduce survival and reproduction. This is the first study to simulate wave-driven flooding and the combined impacts of SLR, groundwater rise, and storm waves on seabird colonies. Our results highlight the need for early climate change planning and restoration of higher elevation seabird refugia to prevent low-lying protected islands from becoming ecological traps in the face of rising sea levels.

  20. Will the Effects of Sea-Level Rise Create Ecological Traps for Pacific Island Seabirds?

    PubMed Central

    Reynolds, Michelle H.; Courtot, Karen N.; Berkowitz, Paul; Storlazzi, Curt D.; Moore, Janet; Flint, Elizabeth

    2015-01-01

    More than 18 million seabirds nest on 58 Pacific islands protected within vast U.S. Marine National Monuments (1.9 million km2). However, most of these seabird colonies are on low-elevation islands and sea-level rise (SLR) and accompanying high-water perturbations are predicted to escalate with climate change. To understand how SLR may impact protected islands and insular biodiversity, we modeled inundation and wave-driven flooding of a globally important seabird rookery in the subtropical Pacific. We acquired new high-resolution Digital Elevation Models (DEMs) and used the Delft3D wave model and ArcGIS to model wave heights and inundation for a range of SLR scenarios (+0.5, +1.0, +1.5, and +2.0 m) at Midway Atoll. Next, we classified vegetation to delineate habitat exposure to inundation and identified how breeding phenology, colony synchrony, and life history traits affect species-specific sensitivity. We identified 3 of 13 species as highly vulnerable to SLR in the Hawaiian Islands and quantified their atoll-wide distribution (Laysan albatross, Phoebastria immutabilis; black-footed albatross, P. nigripes; and Bonin petrel, Pterodroma hypoleuca). Our models of wave-driven flooding forecast nest losses up to 10% greater than passive inundation models at +1.0 m SLR. At projections of + 2.0 m SLR, approximately 60% of albatross and 44% of Bonin petrel nests were overwashed displacing more than 616,400 breeding albatrosses and petrels. Habitat loss due to passive SLR may decrease the carrying capacity of some islands to support seabird colonies, while sudden high-water events directly reduce survival and reproduction. This is the first study to simulate wave-driven flooding and the combined impacts of SLR, groundwater rise, and storm waves on seabird colonies. Our results highlight the need for early climate change planning and restoration of higher elevation seabird refugia to prevent low-lying protected islands from becoming ecological traps in the face of rising sea levels. PMID:26398209

  1. Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins.

    PubMed

    Ast, Cindy; Foret, Jessica; Oltrogge, Luke M; De Michele, Roberto; Kleist, Thomas J; Ho, Cheng-Hsun; Frommer, Wolf B

    2017-09-05

    Sensitivity, dynamic and detection range as well as exclusion of expression and instrumental artifacts are critical for the quantitation of data obtained with fluorescent protein (FP)-based biosensors in vivo. Current biosensors designs are, in general, unable to simultaneously meet all these criteria. Here, we describe a generalizable platform to create dual-FP biosensors with large dynamic ranges by employing a single FP-cassette, named GO-(Green-Orange) Matryoshka. The cassette nests a stable reference FP (large Stokes shift LSSmOrange) within a reporter FP (circularly permuted green FP). GO- Matryoshka yields green and orange fluorescence upon blue excitation. As proof of concept, we converted existing, single-emission biosensors into a series of ratiometric calcium sensors (MatryoshCaMP6s) and ammonium transport activity sensors (AmTryoshka1;3). We additionally identified the internal acid-base equilibrium as a key determinant of the GCaMP dynamic range. Matryoshka technology promises flexibility in the design of a wide spectrum of ratiometric biosensors and expanded in vivo applications.Single fluorescent protein biosensors are susceptible to expression and instrumental artifacts. Here Ast et al. describe a dual fluorescent protein design whereby a reference fluorescent protein is nested within a reporter fluorescent protein to control for such artifacts while preserving sensitivity and dynamic range.

  2. Environmental change mediates mate choice for an extended phenotype, but not for mate quality.

    PubMed

    Head, Megan L; Fox, Rebecca J; Barber, Iain

    2017-01-01

    Sexual cues, including extended phenotypes, are expected to be reliable indicators of male genetic quality and/or provide information on parental quality. However, the reliability of these cues may be dependent on stability of the environment, with heterogeneity affecting how selection acts on such traits. Here, we test how environmental change mediates mate choice for multiple sexual traits, including an extended phenotype--the structure of male-built nests - in stickleback fish. First, we manipulated the dissolved oxygen (DO) content of water to create high or low DO environments in which male fish built nests. Then we recorded the mate choice of females encountering these males (and their nests), under either the same or reversed DO conditions. Males in high DO environments built more compact nests than those in low DO conditions and males adjusted their nest structure in response to changing conditions. Female mate choice for extended phenotype (male nests) was environmentally dependent (females chose more compact nests in high DO conditions), while female choice for male phenotype was not (females chose large, vigorous males regardless of DO level). Examining mate choice in this dynamic context suggests that females evaluate the reliability of multiple sexual cues, taking into account environmental heterogeneity. © 2016 The Author(s). Evolution published by Wiley Periodicals, Inc. on behalf of The Society for the Study of Evolution.

  3. An evaluation and comparison of conservation guidelines for an at-risk migratory songbird

    USGS Publications Warehouse

    McNeil, Darin J.; Aldinger, Kyle R.; Bakermans, Marja H.; Lehman, Justin A.; Tisdale, Anna C.; Jones, John A.; Wood, Petra B.; Buehler, David A.; Smalling, Curtis G.; Siefferman, Lynn; Larkin, Jeffrey L.

    2017-01-01

    For at-risk wildlife species, it is important to consider conservation within the process of adaptive management. Golden-winged Warblers (Vermivora chrysoptera) are Neotropical migratory songbirds that are experiencing long-term population declines due in part to the loss of early-successional nesting habitat. Recently-developed Golden-winged Warbler habitat management guidelines are being implemented by USDA: Natural Resource Conservation Service (2014) and its partners through the Working Lands For Wildlife (WLFW) program. During 2012–2014, we studied the nesting ecology of Golden-winged Warblers in managed habitats of the eastern US that conformed to WLFW conservation practices. We evaluated five NRCS “management scenarios” with respect to nesting success and attainment of recommended nest site vegetation conditions outlined in the Golden-winged Warbler breeding habitat guidelines. Using estimates of territory density, pairing rate, nest survival, and clutch size, we also estimated fledgling productivity (number of fledglings/ha) for each management scenario. In general, Golden-winged Warbler nest survival declined as each breeding season advanced, but nest survival was similar across management scenarios. Within each management scenario, vegetation variables had little influence on nest survival. Still, percent Rubus cover and density of >2 m tall shrubs were relevant in some management scenarios. All five management scenarios rarely attained recommended levels of nest site vegetation conditions for Golden-winged, yet nest survival was high. Fledgling productivity estimates for each management scenario ranged from 2.1 to 8.6 fledglings/10 hectares. Our results indicate that targeted habitat management for Golden-winged Warblers using a variety of management techniques on private lands has the capability to yield high nest survival and fledgling productivity, and thus have the potential to contribute to the species recovery.

  4. The development of loop-mediated isothermal amplification targeting alpha-tubulin DNA for the rapid detection of Plasmodium vivax.

    PubMed

    Dinzouna-Boutamba, Sylvatrie-Danne; Yang, Hye-Won; Joo, So-Young; Jeong, Sookwan; Na, Byoung-Kuk; Inoue, Noboru; Lee, Won-Ki; Kong, Hyun-Hee; Chung, Dong-Il; Goo, Youn-Kyoung; Hong, Yeonchul

    2014-06-30

    Malaria that is caused by Plasmodium vivax is the most widely distributed human malaria. Its recent resurgence in many parts of the world, including the Republic of Korea (ROK), emphasizes the importance of improved access to the early and accurate detection of P. vivax to reduce disease burden. In this study, a rapid and efficient loop-mediated isothermal amplification (LAMP)-based method was developed and validated using blood samples from malaria-suspected patients. A LAMP assay targeting the α-tubulin gene for the detection of P. vivax was developed with six primers that recognize different regions of the target gene. The diagnostic performance of the α-tubulin LAMP assay was compared to three other tests: microscopic examinations, rapid diagnostic tests (RDTs), and nested polymerase chain reactions (PCRs) using 177 whole blood specimens obtained from ROK military personnel from May to December 2011. The α-tubulin LAMP assay was highly sensitive with a detection limit of 100 copies of P. vivax α-tubulin gene per reaction within 50 min. It specifically amplified the target gene only from P. vivax. Validation of the α-tubulin LAMP assay showed that the assay had the highest sensitivity (P < 0.001 versus microscopy; P = 0.0023 versus RDT) when nested PCR was used as the gold standard and better agreement (concordance: 94.9%, kappa value: 0.865) with nested PCR than RDT and microscopy. A Receiver Operation Characteristics analysis showed that the diagnostic accuracy of the α-tubulin LAMP assay for vivax malaria was higher (Area Under Curve = 0.908) than RDT and microscopy. This study showed that the P. vivax α-tubulin LAMP assay, which can be used to diagnose early infections of vivax malaria, is an alternative molecular diagnostic tool and a point-of-care test that may help to prevent transmission in endemic areas.

  5. Breeding loggerhead marine turtles Caretta caretta in Dry Tortugas National Park, USA, show high fidelity to diverse habitats near nesting beaches

    USGS Publications Warehouse

    Hart, Kristen M.; Zawada, David G.; Sartain-Iverson, Autumn R.; Fujisaki, Ikuko

    2016-01-01

    We used satellite telemetry to identify in-water habitat used by individuals in the smallest North-west Atlantic subpopulation of adult nesting loggerhead turtles Caretta caretta during the breeding season. During 2010, 2011 and 2012 breeding periods, a total of 20 adult females used habitats proximal to nesting beaches with various levels of protection within Dry Tortugas National Park. We then used a rapid, high-resolution, digital imaging system to map habitat adjacent to nesting beaches, revealing the diversity and distribution of available benthic cover. Turtle behaviour showing measurable site-fidelity to these diverse habitats has implications for managing protected areas and human activities within them. Protecting diverse benthic areas adjacent to loggerhead turtle nesting beaches here and elsewhere could provide benefits for overall biodiversity conservation.

  6. Mourning Dove nesting habitat and nest success in Central Missouri

    USGS Publications Warehouse

    Drobney, R.D.; Schulz, J.H.; Sheriff, S.L.; Fuemmeler, W.J.

    1998-01-01

    Previous Mourning Dove (Zenaida macroura) nesting studies conducted in areas containing a mixture of edge and continuous habitats have focused on edge habitats. Consequently, little is known about the potential contribution of continuous habitats to dove production. In this study we evaluated the relative importance of these two extensive habitat types by monitoring the habitat use and nest success of 59 radio-marked doves during 1990-1991 in central Missouri. Of 83 nests initiated by our marked sample, most (81.9%) were located in edge habitats. Although continuous habitats were selected less as nest sites, the proportion of successful nests did not differ significantly from that in edge habitats. Our data indicate that continuous habitats should not be considered marginal nesting habitat. If the intensity of use and nest success that we observed are representative regionally or nationally, continuous habitats could contribute substantially to annual Mourning Dove production because of the high availability of these habitats throughout much of the Mourning Dove breeding range.

  7. Development and clinical performance of high throughput loop-mediated isothermal amplification for detection of malaria

    PubMed Central

    Perera, Rushini S.; Ding, Xavier C.; Tully, Frank; Oliver, James; Bright, Nigel; Bell, David; Chiodini, Peter L.; Gonzalez, Iveth J.; Polley, Spencer D.

    2017-01-01

    Background Accurate and efficient detection of sub-microscopic malaria infections is crucial for enabling rapid treatment and interruption of transmission. Commercially available malaria LAMP kits have excellent diagnostic performance, though throughput is limited by the need to prepare samples individually. Here, we evaluate the clinical performance of a newly developed high throughput (HTP) sample processing system for use in conjunction with the Eiken malaria LAMP kit. Methods The HTP system utilised dried blood spots (DBS) and liquid whole blood (WB), with parallel sample processing of 94 samples per run. The system was evaluated using 699 samples of known infection status pre-determined by gold standard nested PCR. Results The sensitivity and specificity of WB-HTP-LAMP was 98.6% (95% CI, 95.7–100), and 99.7% (95% CI, 99.2–100); sensitivity of DBS-HTP-LAMP was 97.1% (95% CI, 93.1–100), and specificity 100% against PCR. At parasite densities greater or equal to 2 parasites/μL, WB and DBS HTP-LAMP showed 100% sensitivity and specificity against PCR. At densities less than 2 p/μL, WB-HTP-LAMP sensitivity was 88.9% (95% CI, 77.1–100) and specificity was 99.7% (95% CI, 99.2–100); sensitivity and specificity of DBS-HTP-LAMP was 77.8% (95% CI, 54.3–99.5) and 100% respectively. Conclusions The HTP-LAMP system is a highly sensitive diagnostic test, with the potential to allow large scale population screening in malaria elimination campaigns. PMID:28166235

  8. Sensitivity of the Carolina Coastal Ocean Circulation to Open Boundary and Atmospheric Forcing

    NASA Astrophysics Data System (ADS)

    Liu, X.; Xie, L.; Pietrafesa, L.

    2003-12-01

    The ocean circulation on the continental shelf off the Carolina coast is characterized by a complex flow regime and temporal variability, which is influenced by atmospheric forcing, the Gulf Stream system, complex coastline and bathymetry, river discharge and tidal forcing. In this study, a triple-nested, HYbrid Coordinate Ocean Model (HYCOM) is used to simulate the coastal ocean circulation on the continental shelf off the Carolina coast and its interactions with the offshore large-scale ocean circulation system. The horizontal mesh size in the innermost domain was set to 1 km, whereas the outermost domain coincides with the near real-time 1/12­’ Atlantic HYCOM Nowcast/Forecast System operated at the Naval Research Laboratory. The intermediate domain uses a mesh size of 3 km. Atmospheric forcing fields for the Carolina coastal region are derived from the NOAA operational ETA model, the ECMWF reanalysis fields and NCEP/NCAR reanalysis fields. These forcing fields are derived at 0.8›¦, 1.125›¦ and 1.875›¦ resolutions, and at intervals of 6 hour, daily and monthly. The sensitivity of the model results to the spatial and temporal resolution of the atmospheric forcing fields is analyzed. To study the dependence of the model sensitivity on the model grid size, single-window simulations at resolutions of 1km, 3km and 9km are carried out using the same forcing fields that were applied to the nested system. Comparisons between the nested and the single domain simulation results will be presented.

  9. Double-Crested Cormorant ( Phalacrocorax auritus) Nesting Effects on Understory Composition and Diversity on Island Ecosystems in Lake Erie

    NASA Astrophysics Data System (ADS)

    McGrath, Darby M.; Murphy, Stephen D.

    2012-08-01

    The context for this study is the management concerns over the severity and extent of the impact of cormorants on island flora in the recent past on Lake Erie islands. Accordingly, this study sought to quantify the nesting colonies' influence on coarse woody litter and how nest densities and litter depth may influence the herbaceous layer, the seed bank composition and viability across the extent of three Lake Erie islands. The data for this study were collected from 2004 to 2008 on East Sister Island and Middle Island using two main strategies. First, herbaceous layer surveys, cormorant nest counts, soil seed bank cores, and litter depth measurements were executed using a plotless-point quarter method to test island-wide impacts from nesting activities (data were also collected on a third island, West Sister Island as a reference for the other two islands). Secondly, a sub-sample of the entire plot set was examined in particularly high nesting density areas for two islands (Middle Island and East Sister Island). Kruskal-Wallis tests indicated that there are subtle changes in the herbaceous diversity (total, native and exotic) and seed bank composition across the islands. The sub sample set of the plots demonstrated that Phalacrocorax auritus nest density does influence litter depth, herbaceous species abundance and diversity. Cormorant nesting pressures are restricted to areas of high nesting pressures and competition. However, there remains a risk to the interior herbaceous layer of the island if the effects of nesting pressures at the edges advance inward from this perimeter.

  10. Variation in spatial scale of competing polydomous twig-nesting ants in coffee agroecosystems

    PubMed Central

    Mathis, Kaitlyn A.; Philpott, Stacy M.; Ramirez, Santiago R.

    2016-01-01

    Arboreal ants are both highly diverse and ecologically dominant in the tropics. This ecologically important group is particularly useful in ongoing efforts to understand processes that regulate species diversity and coexistence. Our study addresses how polydomy can influence patterns of nest occupation in competing arboreal ants. We examined the spatial structure of nest occupation (nest distance, abundance and density) in three polydomous co-occurring twig-nesting ant species (Pseudomyrmex simplex, P. ejectus and P. PSW-53) by mapping twigs occupied by ants from each species within plots in our study site. We then used two colony structure estimators (intraspecific aggression and cuticular hydrocarbon variation) to determine the relative degree of polydomy for each species. All work was conducted in coffee agroforests in Chiapas, Mexico. Our results revealed that the two species with highest abundance and nest density were also highly polydomous, where both species had either single or multiple non-aggressive colonies occupying nests on a large spatial scale (greater than the hectare level). Our results also indicate that the species with the lowest abundance and density is less polydomous, occupying several overlapping and territorial colonies at the hectare level in which multiple colonies never co-occur on the same host plant. These results contribute evidence that successful coexistence and highly polydomous colony structure may allow ants, through reduced intraspecific aggression, to successfully occupy more nests more densely than ant species that have multiple territorial colonies. Furthermore our study highlights the importance of considering intraspecific interactions when examining community assembly of ants. PMID:27795573

  11. [Nesting habitat characterization for Amazona oratrix (Psittaciformes: Psittacidae) in the Central Pacific, Mexico].

    PubMed

    Monterrubio-Rico, Tiberio C; Álvarez-Jara, Margarito; Tellez-Garcia, Loreno; Tena-Morelos, Carlos

    2014-09-01

    The nesting requirements of the Yellow-headed Parrot (Amazona oratrix) are poorly understood, despite their broad historical distribution, high demand for pet trade and current endangered status. Information concerning their nesting requirements is required in order to design specific restoration and conser- vation actions. To assess this, we studied their nesting ecology in the Central Pacific, Michoacan, Mexico during a ten year period. The analyzed variables ranged from local scale nest site characteristics such as nesting tree species, dimensions, geographic positions, diet and nesting forest patches structure, to large scale features such as vegetation use and climatic variables associated to the nesting tree distributions by an ecological niche model using Maxent. We also evaluated the parrot tolerance to land management regimes, and compared the Pacific nest trees with 18 nest trees recorded in an intensively managed private ranch in Tamaulipas, Gulf of Mexico. Parrots nested in tall trees with canopy level cavities in 92 nest-trees recorded from 11 tree species. The 72.8% of nesting occurred in trees of Astronium graveolens, and Enterolobium cyclocarpum which qualified as key- stone trees. The forests where the parrots nested, presented a maximum of 54 tree species, 50% of which were identified as food source; besides, these areas also had a high abundance of trees used as food supply. The lowest number of tree species and trees to forage occurred in an active cattle ranch, whereas the highest species rich- ness was observed in areas with natural recovery. The nesting cavity entrance height from above ground of the Pacific nesting trees resulted higher than those found in the Gulf of Mexico. We hypothesize that the differences may be attributed to Parrot behavioral differences adapting to differential poaching pressure and cavity avail- ability. Nesting trees were found in six vegetation types; however the parrots preferred conserved and riparian semi-deciduous forest for nesting, with fewer nests in deciduous forest, while nesting in transformed agricultural fields was avoided. The main climatic variables associated with the potential distribution of nests were: mean temperature of wettest quarter, mean diurnal temperature range, and precipitation of wettest month. Suitable cli- matic conditions for the potential presence of nesting trees were present in 61% of the region; however, most of the area consisted of tropical deciduous forests (55.8%), while semi-deciduous tropical forests covered only 17% of the region. These results indicated the importance to conserve semi-deciduous forests as breeding habitats for the Yellow-headed Parrot, and revealed the urgent need to implement conservation and restoration actions. These should include a total ban of land use change in tropical semi-deciduous forest areas, and for selective logging of all keystone tree species; besides, we recommend the establishment of wildlife sanctuaries in important nesting areas, and a series of tropical forest restoration programs in the Central Pacific coast.

  12. Diagnosis of neonatal group B Streptococcus sepsis by nested-PCR of residual urine samples

    PubMed Central

    Cezarino, Bruno Nicolino; Yamamoto, Lidia; Del Negro, Gilda Maria Barbaro; Rocha, Daisy; Okay, Thelma Suely

    2008-01-01

    Group B streptococcus (GBS) remains the most common cause of early-onset sepsis in newborns. Laboratory gold-standard, broth culture methods are highly specific, but lack sensitivity. The aim of this study was to validate a nested-PCR and to determine whether residue volumes of urine samples obtained by non invasive, non sterile methods could be used to confirm neonatal GBS sepsis. The nested-PCR was performed with primers of the major GBS surface antigen. Unavailability of biological samples to perform life supporting exams, as well as others to elucidate the etiology of infections is a frequent problem concerning newborn patients. Nevertheless, we decided to include cases according to strict criteria: newborns had to present with signs and symptoms compatible with GBS infection; at least one of the following biological samples had to be sent for culture: blood, urine, or cerebrospinal fluid; availability of residue volumes of the samples sent for cultures, or of others collected on the day of hospitalization, prior to antibiotic therapy prescription, to be analyzed by PCR; favorable outcome after GBS empiric treatment. In only one newborn GBS infection was confirmed by cultures, while infection was only presumptive in the other three patients (they fulfilled inclusion criteria but were GBS-culture negative). From a total of 12 biological samples (5 blood, 3 CSF and 4 urine specimen), eight were tested by culture methods (2/8 were positive), and 8 were tested by PCR (7/8 were positive), and only 4 samples were simultaneously tested by both methods (1 positive by culture and 3 by PCR). In conclusion, although based on a restricted number of neonates and samples, our results suggest that the proposed nested-PCR might be used to diagnose GBS sepsis as it has successfully amplified the three types of biological samples analyzed (blood, urine and cerebrospinal fluid), and was more sensitive than culture methods as PCR in urine confirmed diagnosis in all four patients. Moreover, PCR has enabled us to use residue volumes of urine samples collected by non invasive, non sterile methods, what is technically adequate as GBS is not part of the normal urine flora, thus avoiding invasive procedures such as suprapubic bladder punction or transurethral catheterization. At the same time, the use of urine instead of blood samples could help preventing newborns blood spoliation. PMID:24031170

  13. Vision and visual navigation in nocturnal insects.

    PubMed

    Warrant, Eric; Dacke, Marie

    2011-01-01

    With their highly sensitive visual systems, nocturnal insects have evolved a remarkable capacity to discriminate colors, orient themselves using faint celestial cues, fly unimpeded through a complicated habitat, and navigate to and from a nest using learned visual landmarks. Even though the compound eyes of nocturnal insects are significantly more sensitive to light than those of their closely related diurnal relatives, their photoreceptors absorb photons at very low rates in dim light, even during demanding nocturnal visual tasks. To explain this apparent paradox, it is hypothesized that the necessary bridge between retinal signaling and visual behavior is a neural strategy of spatial and temporal summation at a higher level in the visual system. Exactly where in the visual system this summation takes place, and the nature of the neural circuitry that is involved, is currently unknown but provides a promising avenue for future research.

  14. Nest construction by a ground-nesting bird represents a potential trade-off between egg crypticity and thermoregulation.

    PubMed

    Mayer, Paul M; Smith, Levica M; Ford, Robert G; Watterson, Dustin C; McCutchen, Marshall D; Ryan, Mark R

    2009-04-01

    Predation selects against conspicuous colors in bird eggs and nests, while thermoregulatory constraints select for nest-building behavior that regulates incubation temperatures. We present results that suggest a trade-off between nest crypticity and thermoregulation of eggs based on selection of nest materials by piping plovers (Charadrius melodus), a ground-nesting bird that constructs simple, pebble-lined nests highly vulnerable to predators and exposed to temperature extremes. Piping plovers selected pebbles that were whiter and appeared closer in color to eggs than randomly available pebbles, suggesting a crypsis function. However, nests that were more contrasting in color to surrounding substrates were at greater risk of predation, suggesting an alternate strategy driving selection of white rocks. Near-infrared reflectance of nest pebbles was higher than randomly available pebbles, indicating a direct physical mechanism for heat control through pebble selection. Artificial nests constructed of randomly available pebbles heated more quickly and conferred heat to model eggs, causing eggs to heat more rapidly than in nests constructed from piping plover nest pebbles. Thermal models and field data indicated that temperatures inside nests may remain up to 2-6 degrees C cooler than surrounding substrates. Thermal models indicated that nests heat especially rapidly if not incubated, suggesting that nest construction behavior may serve to keep eggs cooler during the unattended laying period. Thus, pebble selection suggests a potential trade-off between maximizing heat reflectance to improve egg microclimate and minimizing conspicuous contrast of nests with the surrounding substrate to conceal eggs from predators. Nest construction behavior that employs light-colored, thermally reflective materials may represent an evolutionary response by birds and other egg-laying organisms to egg predation and heat stress.

  15. Methods for reducing ghost rays on the Wolter-I focusing figures of the FOXSI rocket payload

    NASA Astrophysics Data System (ADS)

    Buitrago-Casas, Juan Camilo; Glesener, Lindsay; Christe, Steven; Ramsey, Brian; Elsner, Ronald; Courtade, Sasha; Vievering, Juliana; Subramania, Athiray; Krucker, Sam; Bale, Stuart

    2017-08-01

    In high energy solar astrophysics, imaging hard X-rays by direct focusing offers higher dynamic range and greater sensitivity compared to past techniques that used indirect imaging. The Focusing Optics X-ray Solar Imager (FOXSI) is a sounding rocket payload which uses seven sets of nested Wolter-I figured mirrors that, together with seven high-sensitive semiconductor detectors, observes the Sun in hard X-rays by direct focusing. The FOXSI rocket has successfully flown twice and is funded to fly a third time in summer 2018.The Wolter-I geometry consists of two consecutive mirrors, one paraboloid, and one hyperboloid, that reflect photons at grazing angles. Correctly focused X-rays reflect twice, once per mirror segment. For extended sources, like the Sun, off-axis photons at certain incident angles can reflect on only one mirror and still reach the focal plane, generating a pattern of single-bounce photons, or ‘ghost rays’ that can limit the sensitivity of the observation of focused X-rays. Understanding and cutting down the ghost rays on the FOXSI optics will maximize the instrument’s sensitivity of the solar faintest sources for future flights. We present an analysis of the FOXSI ghost rays based on ray-tracing simulations, as well as the effectiveness of different physical strategies to reduce them.

  16. Methods for Reducing Singly Reflected Rays on the Wolter-I Focusing Figures of the FOXSI Rocket Experiment

    NASA Technical Reports Server (NTRS)

    Buitrago-Casas, Juan Camilo; Glesener, Lindsay; Christe, Steven; Elsner, Ronald; Ramsey, Brian; Courtade, Sasha; Ishikawa, Shin-nosuke; Narukage, Noriyuki; Vievering, Juliana; Subramania, Athiray; hide

    2017-01-01

    In high energy solar astrophysics, imaging hard X-rays by direct focusing offers higher dynamic range and greater sensitivity compared to past techniques that used indirect imaging. The Focusing Optics X-ray Solar Imager (FOXSI) is a sounding rocket payload which uses seven sets of nested Wolter-I figured mirrors that, together with seven high-sensitivity semiconductor detectors, observes the Sun in hard X-rays by direct focusing. The FOXSI rocket has successfully flown twice and is funded to fly a third time in Summer 2018. The Wolter-I geometry consists of two consecutive mirrors, one paraboloid, and one hyperboloid, that reflect photons at grazing angles. Correctly focused X-rays reflect twice, once per mirror segment. For extended sources, like the Sun, off-axis photons at certain incident angles can reflect on only one mirror and still reach the focal plane, generating a pattern of single-bounce photons that can limit the sensitivity of the observation of faint focused X-rays. Understanding and cutting down the singly reflected rays on the FOXSI optics will maximize the instrument's sensitivity of the faintest solar sources for future flights. We present an analysis of the FOXSI singly reflected rays based on ray-tracing simulations, as well as the effectiveness of different physical strategies to reduce them.

  17. Development of auditory sensitivity in budgerigars (Melopsittacus undulatus)

    NASA Astrophysics Data System (ADS)

    Brittan-Powell, Elizabeth F.; Dooling, Robert J.

    2004-06-01

    Auditory feedback influences the development of vocalizations in songbirds and parrots; however, little is known about the development of hearing in these birds. The auditory brainstem response was used to track the development of auditory sensitivity in budgerigars from hatch to 6 weeks of age. Responses were first obtained from 1-week-old at high stimulation levels at frequencies at or below 2 kHz, showing that budgerigars do not hear well at hatch. Over the next week, thresholds improved markedly, and responses were obtained for almost all test frequencies throughout the range of hearing by 14 days. By 3 weeks posthatch, birds' best sensitivity shifted from 2 to 2.86 kHz, and the shape of the auditory brainstem response (ABR) audiogram became similar to that of adult budgerigars. About a week before leaving the nest, ABR audiograms of young budgerigars are very similar to those of adult birds. These data complement what is known about vocal development in budgerigars and show that hearing is fully developed by the time that vocal learning begins.

  18. Shrub Nesting of the Red-Eyed Vireo in Relation to Structure of Aspen Forests

    Treesearch

    William J. Jr Mattson

    1979-01-01

    Nests were built in five species of high shrubs in four trembling aspen forests of northern Minnesoa. Nest densities ranged between 0.5 and 3.0 per acre and were positively related to the abundance3 of large (>1.2-cm diameter) shrub stems.

  19. Presence of Breeding Birds Improves Body Condition for a Crocodilian Nest Protector

    PubMed Central

    Nell, Lucas A.; Frederick, Peter C.; Mazzotti, Frank J.; Vliet, Kent A.; Brandt, Laura A.

    2016-01-01

    Ecological associations where one species enhances habitat for another nearby species (facilitations) shape fundamental community dynamics and can promote niche expansion, thereby influencing how and where species persist and coexist. For the many breeding birds facing high nest-predation pressure, enemy-free space can be gained by nesting near more formidable animals for physical protection. While the benefits to protected species seem well documented, very few studies have explored whether and how protector species are affected by nest protection associations. Long-legged wading birds (Pelecaniformes and Ciconiiformes) actively choose nesting sites above resident American alligators (Alligator mississippiensis), apparently to take advantage of the protection from mammalian nest predators that alligator presence offers. Previous research has shown that wading bird nesting colonies could provide substantial food for alligators in the form of dropped chicks. We compared alligator body condition in similar habitat with and without wading bird nesting colonies present. Alligator morphometric body condition indices were significantly higher in colony than in non-colony locations, an effect that was statistically independent of a range of environmental variables. Since colonially nesting birds and crocodilians co-occur in many tropical and subtropical wetlands, our results highlight a potentially widespread keystone process between two ecologically important species-groups. These findings suggest the interaction is highly beneficial for both groups of actors, and illustrate how selective pressures may have acted to form and reinforce a strongly positive ecological interaction. PMID:26934602

  20. Lifespan Analyses of Forest Raptor Nests: Patterns of Creation, Persistence and Reuse

    PubMed Central

    Jiménez-Franco, María V.; Martínez, José E.; Calvo, José F.

    2014-01-01

    Structural elements for breeding such as nests are key resources for the conservation of bird populations. This is especially true when structural elements require a specific and restricted habitat, or if the construction of nests is costly in time and energy. The availability of nesting-platforms is influenced by nest creation and persistence. In a Mediterranean forest in southeastern Spain, nesting-platforms are the only structural element for three forest-dwelling raptor species: booted eagle Aquila pennata, common buzzard Buteo buteo and northern goshawk Accipiter gentilis. From 1998 to 2013, we tracked the fate of 157 nesting-platforms built and reused by these species with the aim of determining the rates of creation and destruction of nesting-platforms, estimating nest persistence by applying two survival analyses, describing the pattern of nest reuse and testing the effects of nest use on breeding success. Nest creation and destruction rates were low (0.14 and 0.05, respectively). Using Kaplan Meier survival estimates and Cox proportional-hazards regression models we found that median nest longevity was 12 years and that this was not significantly affected by nest characteristics, nest-tree dimensions, nest-builder species, or frequency of use of the platform. We also estimated a transition matrix, considering the different stages of nest occupation (vacant or occupied by one of the focal species), to obtain the fundamental matrix and the average life expectancies of nests, which varied from 17.9 to 19.7 years. Eighty six percent of nests were used in at least one breeding attempt, 67.5% were reused and 17.8% were successively occupied by at least two of the study species. The frequency of nest use had no significant effects on the breeding success of any species. We conclude that nesting-platforms constitute an important resource for forest raptors and that their longevity is sufficiently high to allow their reuse in multiple breeding attempts. PMID:24717935

  1. Lifespan analyses of forest raptor nests: patterns of creation, persistence and reuse.

    PubMed

    Jiménez-Franco, María V; Martínez, José E; Calvo, José F

    2014-01-01

    Structural elements for breeding such as nests are key resources for the conservation of bird populations. This is especially true when structural elements require a specific and restricted habitat, or if the construction of nests is costly in time and energy. The availability of nesting-platforms is influenced by nest creation and persistence. In a Mediterranean forest in southeastern Spain, nesting-platforms are the only structural element for three forest-dwelling raptor species: booted eagle Aquila pennata, common buzzard Buteo buteo and northern goshawk Accipiter gentilis. From 1998 to 2013, we tracked the fate of 157 nesting-platforms built and reused by these species with the aim of determining the rates of creation and destruction of nesting-platforms, estimating nest persistence by applying two survival analyses, describing the pattern of nest reuse and testing the effects of nest use on breeding success. Nest creation and destruction rates were low (0.14 and 0.05, respectively). Using Kaplan Meier survival estimates and Cox proportional-hazards regression models we found that median nest longevity was 12 years and that this was not significantly affected by nest characteristics, nest-tree dimensions, nest-builder species, or frequency of use of the platform. We also estimated a transition matrix, considering the different stages of nest occupation (vacant or occupied by one of the focal species), to obtain the fundamental matrix and the average life expectancies of nests, which varied from 17.9 to 19.7 years. Eighty six percent of nests were used in at least one breeding attempt, 67.5% were reused and 17.8% were successively occupied by at least two of the study species. The frequency of nest use had no significant effects on the breeding success of any species. We conclude that nesting-platforms constitute an important resource for forest raptors and that their longevity is sufficiently high to allow their reuse in multiple breeding attempts.

  2. Detection of Talaromyces marneffei from Fresh Tissue of an Inhalational Murine Pulmonary Model Using Nested PCR

    PubMed Central

    Liu, Yinghui; Huang, Xiaowen; Yi, Xiuwen; He, Ya; Mylonakis, Eleftherios; Xi, Liyan

    2016-01-01

    Penicilliosis marneffei, often consecutive to the aspiration of Talaromyces marneffei (Penicillium marneffei), continues to be one of the significant causes of morbidity and mortality in immunocompromised patients in endemic regions such as Southeast Asia. Improving the accuracy of diagnosing this disease would aid in reducing the mortality of associated infections. In this study, we developed a stable and reproducible murine pulmonary model that mimics human penicilliosis marneffei using a nebulizer to deliver Talaromyces marneffei (SUMS0152) conidia to the lungs of BALB/c nude mice housed in exposure chamber. Using this model, we further revealed that nested PCR was sensitive and specific for detecting Talaromyces marneffei in bronchoalveolar lavage fluid and fresh tissues. This inhalation model may provide a more representative analysis tool for studying the development of penicilliosis marneffei, in addition to revealing that nested PCR has a predictive value in reflecting pulmonary infection. PMID:26886887

  3. Estimating population size of Pygoscelid Penguins from TM data

    NASA Technical Reports Server (NTRS)

    Olson, Charles E., Jr.; Schwaller, Mathew R.; Dahmer, Paul A.

    1987-01-01

    An estimate was made toward a continent wide population of penguins. The results indicate that Thematic Mapper data can be used to identify penguin rookeries due to the unique reflectance properties of guano. Strong correlations exist between nesting populations and rookery area occupied by the birds. These correlations allow estimation of the number of nesting pairs in colonies. The success of remote sensing and biometric analyses leads one to believe that a continent wide estimate of penguin populations is possible based on a timely sample employing ground based and remote sensing techniques. Satellite remote sensing along the coastline may well locate previously undiscovered penguin nesting sites, or locate rookeries which have been assumed to exist for over a half century, but never located. Observations which found that penguins are one of the most sensitive elements in the complex of Southern Ocean ecosystems motivated this study.

  4. Does formal complexity reflect cognitive complexity? Investigating aspects of the Chomsky Hierarchy in an artificial language learning study.

    PubMed

    Öttl, Birgit; Jäger, Gerhard; Kaup, Barbara

    2015-01-01

    This study investigated whether formal complexity, as described by the Chomsky Hierarchy, corresponds to cognitive complexity during language learning. According to the Chomsky Hierarchy, nested dependencies (context-free) are less complex than cross-serial dependencies (mildly context-sensitive). In two artificial grammar learning (AGL) experiments participants were presented with a language containing either nested or cross-serial dependencies. A learning effect for both types of dependencies could be observed, but no difference between dependency types emerged. These behavioral findings do not seem to reflect complexity differences as described in the Chomsky Hierarchy. This study extends previous findings in demonstrating learning effects for nested and cross-serial dependencies with more natural stimulus materials in a classical AGL paradigm after only one hour of exposure. The current findings can be taken as a starting point for further exploring the degree to which the Chomsky Hierarchy reflects cognitive processes.

  5. Does Formal Complexity Reflect Cognitive Complexity? Investigating Aspects of the Chomsky Hierarchy in an Artificial Language Learning Study

    PubMed Central

    Öttl, Birgit; Jäger, Gerhard; Kaup, Barbara

    2015-01-01

    This study investigated whether formal complexity, as described by the Chomsky Hierarchy, corresponds to cognitive complexity during language learning. According to the Chomsky Hierarchy, nested dependencies (context-free) are less complex than cross-serial dependencies (mildly context-sensitive). In two artificial grammar learning (AGL) experiments participants were presented with a language containing either nested or cross-serial dependencies. A learning effect for both types of dependencies could be observed, but no difference between dependency types emerged. These behavioral findings do not seem to reflect complexity differences as described in the Chomsky Hierarchy. This study extends previous findings in demonstrating learning effects for nested and cross-serial dependencies with more natural stimulus materials in a classical AGL paradigm after only one hour of exposure. The current findings can be taken as a starting point for further exploring the degree to which the Chomsky Hierarchy reflects cognitive processes. PMID:25885790

  6. Differential effects of coyotes and red foxes on duck nest success

    USGS Publications Warehouse

    Sovada, Marsha A.; Sargeant, A.; Grier, J.W.

    1995-01-01

    Low recruitment rates prevail among ducks in the Prairie Pothole Region of North America, primarily because of high nest depredation rates. The red fox (Vulpes vulpes) is a major predator of duck eggs, but fox abundance is depressed by coyotes (Canis latrans). We tested the hypothesis that nest success of upland-nesting ducks is higher in areas with coyotes than in areas with red foxes. We conducted the study during 1990-92 in uplands of 36 areas managed for nesting ducks in North Dakota and South Dakota. Overall nest success averaged 32% (95% CI = 25-40) on 17 study areas where coyotes were the principal canid and 17% (CI = 11-25) on 13 study areas where red foxes were the principal canid (P = 0.01). Both canids were common on 6 other areas, where nest success averaged 25% (CI = 13-47). Habitat composition, predator communities with the exception of canids, and species composition of duck nests in coyote and red fox areas were similar overall. Upon examining only nests with greater than or equal to 6 eggs on the last visit prior to hatch or depredation, we determined nests with evidence characteristic of fox predation accounted for 4% of depredated nests in coyote areas and 27% in fox areas (P = 0.001). An expanding coyote population is contributing to higher overall nest success. Management of coyotes may be an effective method for increasing duck nest success.

  7. Development of a nested-PCR assay for the rapid detection of Pilidiella granati in pomegranate fruit

    PubMed Central

    Yang, Xue; Hameed, Uzma; Zhang, Ai-Fang; Zang, Hao-Yu; Gu, Chun-Yan; Chen, Yu; Xu, Yi-Liu

    2017-01-01

    Pilidiella granati, a causal agent of twig blight and crown rot of pomegranate, is an emerging threat that may cause severe risk to the pomegranate industry in the future. Development of a rapid assay for the timely and accurate detection of P. granati will be helpful in the active surveillance and management of the disease caused by this pathogen. In this study, a nested PCR method was established for the detection of P. granati. Comparative analysis of genetic diversity within 5.8S rDNA internal transcribed spacer (ITS) sequences of P. granati and 21 other selected fungal species was performed to design species-specific primers (S1 and S2). This primer pair successfully amplified a 450 bp product exclusively from the genomic DNA of P. granati. The developed method can detect 10 pg genomic DNA of the pathogen in about 6 h. This technique was successfully applied to detect the natural infection of P. granati in the pomegranate fruit. The designed protocol is rapid and precise with a high degree of sensitivity. PMID:28106107

  8. How uncertain is the future of electric vehicle market: Results from Monte Carlo simulations using a nested logit model

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Liu, Changzheng; Oak Ridge National Lab.; Lin, Zhenhong

    Plug-in electric vehicles (PEVs) are widely regarded as an important component of the technology portfolio designed to accomplish policy goals in sustainability and energy security. However, the market acceptance of PEVs in the future remains largely uncertain from today's perspective. By integrating a consumer choice model based on nested multinomial logit and Monte Carlo simulation, this study analyzes the uncertainty of PEV market penetration using Monte Carlo simulation. Results suggest that the future market for PEVs is highly uncertain and there is a substantial risk of low penetration in the early and midterm market. Top factors contributing to market sharemore » variability are price sensitivities, energy cost, range limitation, and charging availability. The results also illustrate the potential effect of public policies in promoting PEVs through investment in battery technology and infrastructure deployment. Here, continued improvement of battery technologies and deployment of charging infrastructure alone do not necessarily reduce the spread of market share distributions, but may shift distributions toward right, i.e., increase the probability of having great market success.« less

  9. [Molecular cloning and expression of the severe acute respiratory syndrome-associated coronavirus nucleocapsid protein and its clinical application].

    PubMed

    Lu, Jian; Zhou, Bai-ping; Zhou, Yu-sen; Jiang, Xiao-ling; Wen, Li-xia; Le, Xiao-hua; Li, Bing; Xu, Liu-mei; Li, Li-xiong

    2005-03-01

    To clone and express nucleocapsid (N) protein of the severe acute respiratory syndrome (SARS)-associated coronavirus, and to evaluate its antigenicity and application value in the development of serological diagnostic test for SARS. SARS-associated coronavirus N protein gene was amplified from its genomic RNA by reverse transcript nested polymerase chain reaction (RT-nested-PCR) and cloned into pBAD/Thio-TOPO prokaryotic expression vector. The recombinant N fusion protein was expressed and purified, and its antigenicity and specificity was analyzed by Western Blot, to establish the recombinant N protein-based ELISA for detection of IgG antibodies to SARS-associated coronavirus, and SARS-associated coronavirus lysates-based ELISA was compared parallelly. The recombinant expression vector produced high level of the N fusion protein after induction, and that protein was purified successfully by affinity chromatography and displayed higher antigenicity and specificity as compared with whole virus lysates. The recombinant SARS-associated coronavirus N protein possessed better antigenicity and specificity and could be employed to establish a new, sensitive, and specific ELISA for SARS diagnosis.

  10. How uncertain is the future of electric vehicle market: Results from Monte Carlo simulations using a nested logit model

    DOE PAGES

    Liu, Changzheng; Oak Ridge National Lab.; Lin, Zhenhong; ...

    2016-12-08

    Plug-in electric vehicles (PEVs) are widely regarded as an important component of the technology portfolio designed to accomplish policy goals in sustainability and energy security. However, the market acceptance of PEVs in the future remains largely uncertain from today's perspective. By integrating a consumer choice model based on nested multinomial logit and Monte Carlo simulation, this study analyzes the uncertainty of PEV market penetration using Monte Carlo simulation. Results suggest that the future market for PEVs is highly uncertain and there is a substantial risk of low penetration in the early and midterm market. Top factors contributing to market sharemore » variability are price sensitivities, energy cost, range limitation, and charging availability. The results also illustrate the potential effect of public policies in promoting PEVs through investment in battery technology and infrastructure deployment. Here, continued improvement of battery technologies and deployment of charging infrastructure alone do not necessarily reduce the spread of market share distributions, but may shift distributions toward right, i.e., increase the probability of having great market success.« less

  11. Absence of Asymptomatic Malaria Infection in a Cross-sectional Study in Iranshahr District, Iran under Elimination Programmes.

    PubMed

    Pirahmadi, Sakineh; Zakeri, Sedigheh; Raeisi, Ahmad

    2017-01-01

    Asymptomatic malaria infection provides a reservoir of parasites, causing the persistence of malaria transmission. It accounts an important challenge for successful management of the control, elimination, and eradication programmes in any malaria-endemic region. This investigation was designed to assess the presence and the prevalence of asymptomatic carriers in Iranshahr district of Sistan and Baluchistan Province (2013-2014), with a considerable population movement, during the malaria elimination phase in Iran. Finger-prick blood samples were collected from symptomless (n=250) and febrile (n=50) individuals residing in Iranshahr district, easthern Iran (Hoodian, Mand, Chah-e Giji, Jolgehashem, Esfand, Dalgan and Chahshour) during Jan 2013 to Dec 2014, and Plasmodium infections were detected using light microscopic and highly sensitive nested-PCR techniques. Thick and thin Giemsa-stained blood smears were negative for Plasmodium parasites. In addition, based on nested-PCR analysis, no P. vivax , P. falciparum, and P. malariae parasites were detected among the studied individuals. Investigation the absence of asymptomatic carriers in Iranshahr district was illustrated and achieving malaria elimination in this area is feasible in a near future.

  12. Age and breeding success related to nest position in a White stork Ciconia ciconia colony

    NASA Astrophysics Data System (ADS)

    Vergara, Pablo; Aguirre, José I.

    2006-11-01

    Coloniality is a breeding system that may produce benefits in terms of breeding success, although these advantages could vary according to factors such as colony size or nest position. We studied breeder's age in relation to nest position (peripheral or central) within the colony. In addition, we studied the relationship between breeding success and nest position, controlling for breeder's age, a highly correlated factor, in a White Stork Ciconia ciconia colony over a 7-year period. Our results show that central nests are mainly occupied by adult birds and had lower failure rates. However, controlling for breeder's age, nest position per se did not explain breeding success. The scarce predation and the lack of human disturbance in the study colony could explain the absence of differences in breeding success between different nest positions within the colony.

  13. Red-cockaded Woodpecker Picoides borealis Microhabitat Characteristics and Reproductive Success in a Loblolly-Shortleaf Pine Forest

    USGS Publications Warehouse

    Wood, Douglas R.; Burger, L. Wesley; Vilella, Francisco

    2014-01-01

    We investigated the relationship between red-cockaded woodpecker (Picoides borealis) reproductive success and microhabitat characteristics in a southeastern loblolly (Pinus taeda) and shortleaf (P. echinata) pine forest. From 1997 to 1999, we recorded reproductive success parameters of 41 red-cockaded woodpecker groups at the Bienville National Forest, Mississippi. Microhabitat characteristics were measured for each group during the nesting season. Logistic regression identified understory vegetation height and small nesting season home range size as predictors of red-cockaded woodpecker nest attempts. Linear regression models identified several variables as predictors of red-cockaded woodpecker reproductive success including group density, reduced hardwood component, small nesting season home range size, and shorter foraging distances. Red-cockaded woodpecker reproductive success was correlated with habitat and behavioral characteristics that emphasize high quality habitat. By providing high quality foraging habitat during the nesting season, red-cockaded woodpeckers can successfully reproduce within small home ranges.

  14. Habitat suitability and nest survival of white-headed woodpeckers in unburned forests of Oregon

    USGS Publications Warehouse

    Hollenbeck, Jeff P.; Saab, Victoria A.; Frenzel, Richard W.

    2011-01-01

    We evaluated habitat suitability and nest survival of breeding white-headed woodpeckers (Picoides albolarvatus) in unburned forests of central Oregon, USA. Daily nest-survival rate was positively related to maximum daily temperature during the nest interval and to density of large-diameter trees surrounding the nest tree. We developed a niche-based habitat suitability model (partitioned Mahalanobis distance) for nesting white-headed woodpeckers using remotely sensed data. Along with low elevation, high density of large trees, and low slope, our habitat suitability model suggested that interspersion–juxtaposition of low- and high-canopy cover ponderosa pine (Pinus ponderosa) patches was important for nest-site suitability. Cross-validation suggested the model performed adequately for management planning at a scale >1 ha. Evaluation of mapped habitat suitability index (HSI) suggested that the maximum predictive gain (HSI = 0.36), where the number of nest locations are maximized in the smallest proportion of the modeled landscape, provided an objective initial threshold for identification of suitable habitat. However, managers can choose the threshold HSI most appropriate for their purposes (e.g., locating regions of low–moderate suitability that have potential for habitat restoration). Consequently, our habitat suitability model may be useful for managing dry coniferous forests for white-headed woodpeckers in central Oregon; however, model validation is necessary before our model could be applied to other locations.

  15. Changes in gonadal and adrenal steroid levels in the leatherback sea turtle (Dermochelys coriacea) during the nesting cycle.

    PubMed

    Rostal, D C; Grumbles, J S; Palmer, K S; Lance, V A; Spotila, J R; Paladino, F V

    2001-05-01

    The reproductive endocrinology of nesting leatherback turtles (Dermochelys coriacea) was studied during the 1996-1997 and 1997-1998 nesting seasons at Los Baulas National Park, Playa Grande, Costa Rica. Blood samples were collected from nesting females throughout the season. Females were observed to nest up to 10 times during the nesting season. Plasma steroids were measured by radioimmunoassay and total plasma calcium was measured by flame atomic absorption spectrophotometry. Plasma testosterone and plasma estradiol levels declined throughout the nesting cycle of the female. Testosterone declined from a mean of 10.18 +/- 0.77 ng/ml at the beginning of the nesting cycle to 1.73 +/- 0.34 ng/ml at the end of the nesting cycle. Estradiol declined in a similar manner, ranging from a mean of 190.95 +/- 16.80 pg/ml at the beginning of the nesting cycle to 76.52 +/- 12.66 pg/ml at the end of the nesting cycle. Plasma progesterone and total calcium levels were relatively constant throughout the nesting cycle. Lack of fluctuation of total calcium levels, ranging from a mean high of 97.46 +/- 11.37 microg/ml to a mean low of 64.85 +/- 11.20 microg/ml, further suggests that vitellogenesis is complete prior to the arrival of the female at the nesting beach. Clutch size (both yolked and yolkless eggs) did not vary over the course of the nesting cycle. Copyright 2001 Academic Press.

  16. Comparison of PCR and culture to the indirect fluorescent-antibody test for diagnosis of Potomac horse fever.

    PubMed

    Mott, J; Rikihisa, Y; Zhang, Y; Reed, S M; Yu, C Y

    1997-09-01

    Potomac horse fever is an acute systemic equine disease caused by Ehrlichia risticii. Currently, serologic methods are widely used to diagnose this disease. However, serologic methods cannot determine whether the horse is presently infected or has been exposed to ehrlichial antigens in the past. The purpose of the present study was to compare the sensitivities of the nested PCR and cell culture with that of the indirect fluorescent-antibody (IFA) test for the diagnosis of Potomac horse fever. Blood and fecal specimens serially collected from a pony experimentally infected with E. risticii Maryland, blood specimens serially collected from mice inoculated with E. risticii Ohio 380, and blood and/or fecal specimens collected from 27 horses which had clinical signs compatible with Potomac horse fever were examined. These horses resided in Kentucky, Indiana, Pennsylvania, and Vermont. The IFA test titer became positive after 6 days postinoculation (p.i.) for the pony. A culture of the blood of the pony was positive for E. risticii starting on day 1 and was positive through day 28 p.i. By the nested PCR, E. risticii was detectable in the blood and feces of the pony starting on day 1 and was detectable through day 32 p.i. E. risticii was detected in the blood of subclinically infected mice by the nested PCR. Twenty-two clinical specimens were seropositive for E. risticii by the IFA test, with titers ranging from 1:20 to 1:1,280. E. risticii was cultured from 95% (20 of 21) of seropositive clinical blood specimens. E. risticii was detected in the blood by PCR in 81% (17 of 20) of the culture-positive clinical specimens. The study indicated that the nested PCR is as sensitive as culture for detecting infection with E. risticii.

  17. Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR

    PubMed Central

    2012-01-01

    Background Gametocyte carriage is essential for malaria transmission and endemicity of disease; thereby it is a target for malaria control strategies. Malaria-infected individuals may harbour gametocytes below the microscopic detection threshold that can be detected by reverse transcription polymerase chain reaction (RT-PCR) targeting gametocyte-specific mRNA. To date, RT-PCR has mainly been applied to the diagnosis of Plasmodium falciparum gametocytes but very limited for that of Plasmodium vivax. Methods A multiplex-nested RT-PCR targeting Pfs25 and Pvs25 mRNA specific to mature gametocytes of P. falciparum and P. vivax, respectively, was developed. The assay was evaluated using blood samples collected in rainy and dry seasons from febrile patients,in a malaria-endemic area in Thailand. Malaria diagnosis was performed by Giemsa-stained blood smears and 18S rRNA PCR. Results The multiplex-nested RT-PCR detected Pfs25 mRNA in 75 of 86 (87.2%) P. falciparum-infected individuals and Pvs25 mRNA in 82 of 90 (91.1%) P. vivax malaria patients diagnosed by 18S rRNA PCR. Gametocytes were detected in 38 (eight P. falciparum and 30 P. vivax) of 157 microscopy positive samples, implying that a large number of patients harbour sub-microscopic gametocytaemia. No seasonal differences in gametocyte carriage were observed for both malaria species diagnosed by multiplex-nested RT-PCR. With single-nested RT-PCR targeting Pfs25 or Pvs25 mRNA as standard, the multiplex-nested RT-PCR offered sensitivities of 97.4% and 98.9% and specificities of 100% and 98.8% for diagnosing mature gametocytes of P. falciparum and P. vivax, respectively. The minimum detection limit of the multiplex-nested PCR was 10 copies of templates. Conclusions The multiplex-nested RT-PCR developed herein is useful for simultaneous assessment of both P. falciparum and P. vivax gametocyte carriage that is prevalent and generally sympatric in several malaria-endemic areas outside Africa. PMID:22682065

  18. Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR.

    PubMed

    Kuamsab, Napaporn; Putaporntip, Chaturong; Pattanawong, Urassaya; Jongwutiwes, Somchai

    2012-06-10

    Gametocyte carriage is essential for malaria transmission and endemicity of disease; thereby it is a target for malaria control strategies. Malaria-infected individuals may harbour gametocytes below the microscopic detection threshold that can be detected by reverse transcription polymerase chain reaction (RT-PCR) targeting gametocyte-specific mRNA. To date, RT-PCR has mainly been applied to the diagnosis of Plasmodium falciparum gametocytes but very limited for that of Plasmodium vivax. A multiplex-nested RT-PCR targeting Pfs25 and Pvs25 mRNA specific to mature gametocytes of P. falciparum and P. vivax, respectively, was developed. The assay was evaluated using blood samples collected in rainy and dry seasons from febrile patients,in a malaria-endemic area in Thailand. Malaria diagnosis was performed by Giemsa-stained blood smears and 18S rRNA PCR. The multiplex-nested RT-PCR detected Pfs25 mRNA in 75 of 86 (87.2%) P. falciparum-infected individuals and Pvs25 mRNA in 82 of 90 (91.1%) P. vivax malaria patients diagnosed by 18S rRNA PCR. Gametocytes were detected in 38 (eight P. falciparum and 30 P. vivax) of 157 microscopy positive samples, implying that a large number of patients harbour sub-microscopic gametocytaemia. No seasonal differences in gametocyte carriage were observed for both malaria species diagnosed by multiplex-nested RT-PCR. With single-nested RT-PCR targeting Pfs25 or Pvs25 mRNA as standard, the multiplex-nested RT-PCR offered sensitivities of 97.4% and 98.9% and specificities of 100% and 98.8% for diagnosing mature gametocytes of P. falciparum and P. vivax, respectively. The minimum detection limit of the multiplex-nested PCR was 10 copies of templates. The multiplex-nested RT-PCR developed herein is useful for simultaneous assessment of both P. falciparum and P. vivax gametocyte carriage that is prevalent and generally sympatric in several malaria-endemic areas outside Africa.

  19. Effects of climate and exurban development on nest predation and predator presence in the southern Appalachian Mountains (USA).

    PubMed

    Lumpkin, Heather A; Pearson, Scott M; Turner, Monica G

    2012-08-01

    In the eastern United States, land-use and climate change have likely contributed to declines in the abundance of Neotropical migrant birds that occupy forest interiors, but the mechanisms are not well understood. We conducted a nest-predation experiment in southern Appalachian Mountain forests (North Carolina, U.S.A.) during the 2009 and 2010 breeding seasons to determine the effects of exurban development and temperature on predator presence and the average number of days until eggs in an artificial nest were disturbed by predators. We baited artificial nests with quail (Excalfactoria chinensi) eggs and monitored them for 18 days. We used clay eggs, track plates, and motion-triggered cameras to detect and identify nest predators. The average number of days a nest was undisturbed decreased as mean temperature increased and, to a lesser extent, as the density of buildings increased. Nests on the ground were more often depredated than those in trees, likely due to increased predation by opossum (Didelphis virginiana) and other carnivores. Raccoons (Procyon lotor), opossums, corvids (Corvus brachyrhynchos and Cyanocitta cristata), chipmunks (Tamias striatus), black bears (Ursus americanus), and domestic cats (Felis catus) were the most commonly detected predators. Presence of these predators did not vary as a function of mean temperature. Domestic cats and corvids were detected more frequently in plots with high rather than low densities of buildings. Forest-interior specialists and Neotropical migrants often nest in cool, high-elevation areas with low housing density. These bird species, especially those that nest on the ground, may be most vulnerable to increased nest predation if temperature and exurban development increase at higher elevations as anticipated. ©2012 Society for Conservation Biology.

  20. Breeding sex ratio and population size of loggerhead turtles from Southwestern Florida.

    PubMed

    Lasala, Jacob A; Hughes, Colin R; Wyneken, Jeanette

    2018-01-01

    Species that display temperature-dependent sex determination are at risk as a result of increasing global temperatures. For marine turtles, high incubation temperatures can skew sex ratios towards females. There are concerns that temperature increases may result in highly female-biased offspring sex ratios, which would drive a future sex ratio skew. Studying the sex ratios of adults in the ocean is logistically very difficult because individuals are widely distributed and males are inaccessible because they remain in the ocean. Breeding sex ratios (BSR) are sought as a functional alternative to study adult sex ratios. One way to examine BSR is to determine the number of males that contribute to nests. Our goal was to evaluate the BSR for loggerhead turtles (Caretta caretta) nesting along the eastern Gulf of Mexico in Florida, from 2013-2015, encompassing three nesting seasons. We genotyped 64 nesting females (approximately 28% of all turtles nesting at that time) and up to 20 hatchlings from their nests (n = 989) using 7 polymorphic microsatellite markers. We identified multiple paternal contributions in 70% of the nests analyzed and 126 individual males. The breeding sex ratio was approximately 1 female for every 2.5 males. We did not find repeat males in any of our nests. The sex ratio and lack of repeating males was surprising because of female-biased primary sex ratios. We hypothesize that females mate offshore of their nesting beaches as well as en route. We recommend further comparisons of subsequent nesting events and of other beaches as it is imperative to establish baseline breeding sex ratios to understand how growing populations behave before extreme environmental effects are evident.

  1. Breeding sex ratio and population size of loggerhead turtles from Southwestern Florida

    PubMed Central

    Hughes, Colin R.; Wyneken, Jeanette

    2018-01-01

    Species that display temperature-dependent sex determination are at risk as a result of increasing global temperatures. For marine turtles, high incubation temperatures can skew sex ratios towards females. There are concerns that temperature increases may result in highly female-biased offspring sex ratios, which would drive a future sex ratio skew. Studying the sex ratios of adults in the ocean is logistically very difficult because individuals are widely distributed and males are inaccessible because they remain in the ocean. Breeding sex ratios (BSR) are sought as a functional alternative to study adult sex ratios. One way to examine BSR is to determine the number of males that contribute to nests. Our goal was to evaluate the BSR for loggerhead turtles (Caretta caretta) nesting along the eastern Gulf of Mexico in Florida, from 2013–2015, encompassing three nesting seasons. We genotyped 64 nesting females (approximately 28% of all turtles nesting at that time) and up to 20 hatchlings from their nests (n = 989) using 7 polymorphic microsatellite markers. We identified multiple paternal contributions in 70% of the nests analyzed and 126 individual males. The breeding sex ratio was approximately 1 female for every 2.5 males. We did not find repeat males in any of our nests. The sex ratio and lack of repeating males was surprising because of female-biased primary sex ratios. We hypothesize that females mate offshore of their nesting beaches as well as en route. We recommend further comparisons of subsequent nesting events and of other beaches as it is imperative to establish baseline breeding sex ratios to understand how growing populations behave before extreme environmental effects are evident. PMID:29370223

  2. Thermal regimes of Mexican spotted owl nest stands

    Treesearch

    Joseph L. Ganey

    2004-01-01

    To evaluate the hypothesis that spotted owls (Strix occidentalis) select habitats with cool microclimates to avoid high daytime temperatures, I sampled thermal regimes in nest areas used by Mexican spotted owls (S. o. lucida) in northern Arizona. I sampled air temperature at 30-min intervals in 30 pairs of nest and random sites...

  3. Reproductive success and habitat characteristics of Golden-winged Warblers in high-elevation pasturelands

    USGS Publications Warehouse

    Wood, Petra; Aldinger, Kyle R.

    2016-01-01

    The Golden-winged Warbler (Vermivora chrysoptera) is one of the most rapidly declining vertebrate species in the Appalachian Mountains. It is the subject of extensive range-wide research and conservation action. However, little is known about this species' breeding ecology in high-elevation pasturelands, a breeding habitat with conservation potential considering the U.S. Natural Resource Conservation Service's Working Lands for Wildlife program targeting private lands in the Appalachian Mountains. We located 100 nests of Golden-winged Warblers in pastures in and around the Monongahela National Forest in West Virginia during 2008–2012. Daily nest survival rate (mean ± SE  =  0.962 ± 0.006), clutch size (4.5 ± 0.1), and number of young fledged per nest attempt (2.0 ± 0.2) and successful nest (4.0 ± 0.1) fell within the range of values reported in other parts of the species' range and were not significantly affected by year or the presence/absence of cattle grazing. Classification tree analysis revealed that nests were in denser vegetation (≥52%) and closer to forest edges (<36.0 m) and shrubs (<7.0 cm) than random locations within the male's territory. Successful nests had significantly more woody cover (≥9%) within 1 m than failed nests. Our results suggest that cattle grazing at 1.2–2.4 ha of forage/animal unit with periodic mowing can create and maintain these characteristics without interfering with the nesting of Golden-winged Warblers. High-elevation pasturelands may provide a refuge for remaining populations of Golden-winged Warblers in this region.

  4. Biomimetic Ant-Nest Electrode Structures for High Sulfur Ratio Lithium–Sulfur Batteries

    DOE PAGES

    Ai, Guo; Dai, Yiling; Mao, Wenfeng; ...

    2016-08-08

    The lithium–sulfur (Li–S) rechargeable battery has the benefit of high gravimetric energy density and low cost. Significant research currently focuses on increasing the sulfur loading and sulfur/inactive-materials ratio, to improve life and capacity. Inspired by nature’s ant-nest structure, this study results in a novel Li–S electrode that is designed to meet both goals. With only three simple manufacturing-friendly steps, which include slurry ball-milling, doctor-blade-based laminate casting, and the use of the sacrificial method with water to dissolve away table salt, the ant-nest design has been successfully recreated in an Li–S electrode. The efficient capabilities of the ant-nest structure are adoptedmore » to facilitate fast ion transportation, sustain polysulfide dissolution, and assist efficient precipitation. Finally, high cycling stability in the Li–S batteries, for practical applications, has been achieved with up to 3 mg·cm –2 sulfur loading. Li–S electrodes with up to a 85% sulfur ratio have also been achieved for the efficient design of this novel ant-nest structure.« less

  5. Effects of parents and Brown-headed Cowbirds (Molothrus ater) on nest predation risk for a songbird

    PubMed Central

    Latif, Quresh S; Heath, Sacha K; Rotenberry, John T

    2012-01-01

    Nest predation limits avian fitness, so ornithologists study nest predation, but they often only document patterns of predation rates without substantively investigating underlying mechanisms. Parental behavior and predator ecology are two fundamental drivers of predation rates and patterns, but the role of parents is less certain, particularly for songbirds. Previous work reproduced microhabitat-predation patterns experienced by Yellow Warblers (Setophaga petechia) in the Mono Lake basin at experimental nests without parents, suggesting that these patterns were driven by predator ecology rather than predator interactions with parents. In this study, we further explored effects of post-initiation parental behavior (nest defense and attendance) on predation risk by comparing natural versus experimental patterns related to territory density, seasonal timing of nest initiation, and nest age. Rates of parasitism by Brown-headed Cowbirds (Molothrus ater) were high in this system (49% nests parasitized), so we also examined parasitism-predation relationships. Natural nest predation rates (NPR) correlated negatively with breeding territory density and nonlinearly (U-shaped relationship) with nest-initiation timing, but experimental nests recorded no such patterns. After adjusting natural-nest data to control for these differences from experimental nests other than the presence of parents (e.g., defining nest failure similarly and excluding nestling-period data), we obtained similar results. Thus, parents were necessary to produce observed patterns. Lower natural NPR compared with experimental NPR suggested that parents reduced predation rates via nest defense, so this parental behavior or its consequences were likely correlated with density or seasonal timing. In contrast, daily predation rates decreased with nest age for both nest types, indicating this pattern did not involve parents. Parasitized nests suffered higher rates of partial predation but lower rates of complete predation, suggesting direct predation by cowbirds. Explicit behavioral research on parents, predators (including cowbirds), and their interactions would further illuminate mechanisms underlying the density, seasonal, and nest age patterns we observed. PMID:23301174

  6. Nest trampling and ground nesting birds: Quantifying temporal and spatial overlap between cattle activity and breeding redshank.

    PubMed

    Sharps, Elwyn; Smart, Jennifer; Mason, Lucy R; Jones, Kate; Skov, Martin W; Garbutt, Angus; Hiddink, Jan G

    2017-08-01

    Conservation grazing for breeding birds needs to balance the positive effects on vegetation structure and negative effects of nest trampling. In the UK, populations of Common redshank Tringa totanus breeding on saltmarshes declined by >50% between 1985 and 2011. These declines have been linked to changes in grazing management. The highest breeding densities of redshank on saltmarshes are found in lightly grazed areas. Conservation initiatives have encouraged low-intensity grazing at <1 cattle/ha, but even these levels of grazing can result in high levels of nest trampling. If livestock distribution is not spatially or temporally homogenous but concentrated where and when redshank breed, rates of nest trampling may be much higher than expected based on livestock density alone. By GPS tracking cattle on saltmarshes and monitoring trampling of dummy nests, this study quantified (i) the spatial and temporal distribution of cattle in relation to the distribution of redshank nesting habitats and (ii) trampling rates of dummy nests. The distribution of livestock was highly variable depending on both time in the season and the saltmarsh under study, with cattle using between 3% and 42% of the saltmarsh extent and spending most their time on higher elevation habitat within 500 m of the sea wall, but moving further onto the saltmarsh as the season progressed. Breeding redshank also nest on these higher elevation zones, and this breeding coincides with the early period of grazing. Probability of nest trampling was correlated to livestock density and was up to six times higher in the areas where redshank breed. This overlap in both space and time of the habitat use of cattle and redshank means that the trampling probability of a nest can be much higher than would be expected based on standard measures of cattle density. Synthesis and applications : Because saltmarsh grazing is required to maintain a favorable vegetation structure for redshank breeding, grazing management should aim to keep livestock away from redshank nesting habitat between mid-April and mid-July when nests are active, through delaying the onset of grazing or introducing a rotational grazing system.

  7. Why wasp foundresses change nests: relatedness, dominance, and nest quality.

    PubMed

    Seppä, Perttu; Queller, David C; Strassmann, Joan E

    2012-01-01

    The costs and benefits of different social options are best understood when individuals can be followed as they make different choices, something that can be difficult in social insects. In this detailed study, we follow overwintered females of the social wasp Polistes carolina through different nesting strategies in a stratified habitat where nest site quality varies with proximity to a foraging area, and genetic relatedness among females is known. Females may initiate nests, join nests temporarily or permanently, or abandon nests. Females can become helpers or egglayers, effectively workers or queens. What they actually do can be predicted by a combination of ecological and relatedness factors. Advantages through increased lifetime success of individuals and nests drives foundresses of the social wasp Polistes from solitary to social nest founding. We studied reproductive options of spring foundresses of P. carolina by monitoring individually-marked wasps and assessing reproductive success of each foundress by using DNA microsatellites. We examined what behavioral decisions foundresses make after relaxing a strong ecological constraint, shortage of nesting sites. We also look at the reproductive consequences of different behaviors. As in other Polistes, the most successful strategy for a foundress was to initiate a nest as early as possible and then accept others as subordinates. A common feature for many P. carolina foundresses was, however, that they reassessed their reproductive options by actively monitoring other nests at the field site and sometimes moving permanently to new nests should that offer better (inclusive) fitness prospects compared to their original nests. A clear motivation for moving to new nests was high genetic relatedness; by the end of the foundress period all females were on nests with full sisters.

  8. Arboreal ant colonies as 'hot-points' of cryptic diversity for myrmecophiles: the weaver ant Camponotus sp. aff. textor and its interaction network with its associates.

    PubMed

    Pérez-Lachaud, Gabriela; Lachaud, Jean-Paul

    2014-01-01

    Systematic surveys of macrofaunal diversity within ant colonies are lacking, particularly for ants nesting in microhabitats that are difficult to sample. Species associated with ants are generally small and rarely collected organisms, which makes them more likely to be unnoticed. We assumed that this tendency is greater for arthropod communities in microhabitats with low accessibility, such as those found in the nests of arboreal ants that may constitute a source of cryptic biodiversity. We investigated the invertebrate diversity associated with an undescribed, but already threatened, Neotropical Camponotus weaver ant. As most of the common sampling methods used in studies of ant diversity are not suited for evaluating myrmecophile diversity within ant nests, we evaluated the macrofauna within ant nests through exhaustive colony sampling of three nests and examination of more than 80,000 individuals. We identified invertebrates from three classes belonging to 18 taxa, some of which were new to science, and recorded the first instance of the co-occurrence of two brood parasitoid wasp families attacking the same ant host colony. This diversity of ant associates corresponded to a highly complex interaction network. Agonistic interactions prevailed, but the prevalence of myrmecophiles was remarkably low. Our data support the hypothesis of the evolution of low virulence in a variety of symbionts associated with large insect societies. Because most myrmecophiles found in this work are rare, strictly specific, and exhibit highly specialized biology, the risk of extinction for these hitherto unknown invertebrates and their natural enemies is high. The cryptic, far unappreciated diversity within arboreal ant nests in areas at high risk of habitat loss qualifies these nests as 'hot-points' of biodiversity that urgently require special attention as a component of conservation and management programs.

  9. Nesting fidelity and molecular evidence for natal homing in the freshwater turtle, Graptemys kohnii

    PubMed Central

    Freedberg, Steven; Ewert, Michael A; Ridenhour, Benjamin J; Neiman, Maurine; Nelson, Craig E

    2005-01-01

    Numerous studies of sea turtle nesting ecology have revealed that females exhibit natal homing, whereby they imprint on the nesting area from which they hatch and subsequently return there to nest as adults. Because freshwater turtles comprise the majority of reptiles known to display environmental sex determination (ESD), the study of natal homing in this group may shed light on recent evolutionary models of sex allocation that are predicated on natal homing in reptiles with ESD. We examined natal homing in Graptemys kohnii, a freshwater turtle with ESD, using mitochondrial sequencing, microsatellite genotyping and mark and recapture of 290 nesting females. Females showed high fidelity to nesting areas, even after being transplanted several kilometres away. A Mantel test revealed significant genetic isolation by distance with respect to nesting locations (r=0.147; p<0.05), suggesting that related females nest in close proximity to one another. The patterns of fidelity and genotype distributions are consistent with homing at a scale that may affect population sex ratios. PMID:16006324

  10. Effects of habitat on avian productivity in abandoned pecan orchards in southern Georgia

    USGS Publications Warehouse

    Patnode, K.A.; White, D.H.

    1992-01-01

    Daily survival rates (DSRs) of nests, eggs and nestlings were determined for Mourning Doves (Zenaida macroura), Northern Cardinals (Cardinalis cardinalis), Brown Thrashers (Toxostoma rufum) and Northern Mockingbirds (Mimus polyglottos) occupying abandoned pecan orchards in a highly fragmented and intensively farmed area of southern Georgia. The effects of nest placement parameters, seasonal factors and habitat disruptions on DSRs for all species combined were statistically analyzed. Egg and nestling DSRs varied significantly by month of nesting, percent cover, vegetative form and position of nest in substrate. Causes of nest failure (no fledglings produced) in order of decreasing importance were predation by small mammals/snakes, avian predation, predation by large mammals, and abandonment. Results provide further evidence that the importance of nest placement and habitat disruptions in nesting success is influenced by foraging strategies of the predator community. Site-specific predator/habitat complexes may be a more appropriate criterion than habitat conditions alone for evaluating avian nesting habitat

  11. Nested neural networks

    NASA Technical Reports Server (NTRS)

    Baram, Yoram

    1988-01-01

    Nested neural networks, consisting of small interconnected subnetworks, allow for the storage and retrieval of neural state patterns of different sizes. The subnetworks are naturally categorized by layers of corresponding to spatial frequencies in the pattern field. The storage capacity and the error correction capability of the subnetworks generally increase with the degree of connectivity between layers (the nesting degree). Storage of only few subpatterns in each subnetworks results in a vast storage capacity of patterns and subpatterns in the nested network, maintaining high stability and error correction capability.

  12. Anthropogenically-Mediated Density Dependence in a Declining Farmland Bird

    PubMed Central

    Dunn, Jenny C.; Hamer, Keith C.; Benton, Tim G.

    2015-01-01

    Land management intrinsically influences the distribution of animals and can consequently alter the potential for density-dependent processes to act within populations. For declining species, high densities of breeding territories are typically considered to represent productive populations. However, as density-dependent effects of food limitation or predator pressure may occur (especially when species are dependent upon separate nesting and foraging habitats), high territory density may limit per-capita productivity. Here, we use a declining but widespread European farmland bird, the yellowhammer Emberiza citrinella L., as a model system to test whether higher territory densities result in lower fledging success, parental provisioning rates or nestling growth rates compared to lower densities. Organic landscapes held higher territory densities, but nests on organic farms fledged fewer nestlings, translating to a 5 times higher rate of population shrinkage on organic farms compared to conventional. In addition, when parental provisioning behaviour was not restricted by predation risk (i.e., at times of low corvid activity), nestling provisioning rates were higher at lower territory densities, resulting in a much greater increase in nestling mass in low density areas, suggesting that food limitation occurred at high densities. These findings in turn suggest an ecological trap, whereby preferred nesting habitat does not provide sufficient food for rearing nestlings at high population density, creating a population sink. Habitat management for farmland birds should focus not simply on creating a high nesting density, but also on ensuring heterogeneous habitats to provide food resources in close proximity to nesting birds, even if this occurs through potentially restricting overall nest density but increasing population-level breeding success. PMID:26431173

  13. Age-specific productivity and nest site characteristics of Cooper's hawks (Accipiter cooperii)

    USGS Publications Warehouse

    Moore, K.R.; Henny, C.J.

    1984-01-01

    Nesting Cooper's Hawks (Accipiter cooperii) were studied in northeastern Oregon. Second-year (SY) males did not breed, but 22 percent of the breeding females were SY's. Mean clutch size (P = 0.012) and mean number of young fledged per pair that laid eggs (P < 0.10) were lower for SY females than for adult (after second year [ASY}) females; however, an equal percentage of the eggs (excluding a collected sample egg) yielded fledged young for each age class. Stepwise discriminant analysis suggested differences in structural characteristics of the nest site habitat for ASY and SY females, i.e., SY female nest sites were associated with younger successional stages than ASY female nest sites. Nests of both age groups were built in trees with high crown volume, but ASY females utilized mistletoe (Arceuthobium sp.) for nest structures more frequently (P < 0.01) than SY females.

  14. Coupling contaminants with demography: Effects of lead and selenium in Pacific common eiders

    USGS Publications Warehouse

    Wilson, H.M.; Flint, Paul L.; Powell, A.N.

    2007-01-01

    We coupled intensive population monitoring with collection of blood samples from 383 nesting Pacific common eiders (Somateria mollisima v-nigrum) at two locations in Alaska (USA) from 2002 to 2004. We investigated annual, geographic, and within-season variation in blood concentrations of lead and selenium; compared exposure patterns with sympatrically nesting spectacled eiders (Somateria fischeri); and examined relationships with clutch size, egg viability, probability of hatching, and apparent survival of adult females. Lead concentrations were elevated in 3.6% of females, and all individuals exhibited elevated selenium, most (81%) at concentrations associated with death in captive waterfowl. Blood lead and selenium concentrations varied both within and among site-years and were lower than those of spectacled eiders. During incubation, blood lead concentrations in females increased significantly (possibly via re-release of stored lead from bone), whereas selenium concentrations decreased (likely because of natural excretion). Probability of a nest containing at least one nonviable egg was positively related to blood selenium in hens, but adverse effects in other life-history variables were not supported. Although reproduction appeared to be sensitive to selenium toxicity, our data suggest that high rates of nonviability are unlikely in this population and that selenium-related reductions to clutch size would be inconsequential at the scale of overall population dynamics. We conclude that Pacific common eiders and other wild marine birds likely have higher selenium tolerances than freshwater species and that interspecific differences in exposure levels may reflect differences in reproductive strategies.

  15. Modeling tradeoffs in avian life history traits and consequences for population growth

    USGS Publications Warehouse

    Clark, M.E.; Martin, T.E.

    2007-01-01

    Variation in population dynamics is inherently related to life history characteristics of species, which vary markedly even within phylogenetic groups such as passerine birds. We computed the finite rate of population change (??) from a matrix projection model and from mark-recapture observations for 23 bird species breeding in northern Arizona. We used sensitivity analyses and a simulation model to separate contributions of different life history traits to population growth rate. In particular we focused on contrasting effects of components of reproduction (nest success, clutch size, number of clutches, and juvenile survival) versus adult survival on ??. We explored how changes in nest success or adult survival coupled to costs in other life history parameters affected ?? over a life history gradient provided by our 23 Arizona species, as well as a broader sample of 121 North American passerine species. We further examined these effects for more than 200 passeriform and piciform populations breeding across North America. Model simulations indicate nest success and juvenile survival exert the largest effects on population growth in species with moderate to high reproductive output, whereas adult survival contributed more to population growth in long-lived species. Our simulations suggest that monitoring breeding success in populations across a broad geographic area provides an important index for identifying neotropical migratory populations at risk of serious population declines and a potential method for identifying large-scale mechanisms regulating population dynamics. ?? 2007 Elsevier B.V. All rights reserved.

  16. Relative roles of grey squirrels, supplementary feeding, and habitat in shaping urban bird assemblages.

    PubMed

    Bonnington, Colin; Gaston, Kevin J; Evans, Karl L

    2014-01-01

    Non-native species are frequently considered to influence urban assemblages. The grey squirrel Sciurus carolinensis is one such species that is widespread in the UK and is starting to spread across Europe; it predates birds' nests and can compete with birds for supplementary food. Using distance sampling across the urbanisation intensity gradient in Sheffield (UK) we test whether urban grey squirrels influence avian species richness and density through nest predation and competition for supplementary food sources. We also assess how urban bird assemblages respond to supplementary feeding. We find that grey squirrels slightly reduced the abundance of breeding bird species most sensitive to squirrel nest predation by reducing the beneficial impact of woodland cover. There was no evidence that grey squirrel presence altered relationships between supplementary feeding and avian assemblage structure. This may be because, somewhat surprisingly, supplementary feeding was not associated with the richness or density of wintering bird assemblages. These associations were positive during the summer, supporting advocacy to feed birds during the breeding season and not just winter, but explanatory capacity was limited. The amount of green space and its quality, assessed as canopy cover, had a stronger influence on avian species richness and population size than the presence of grey squirrels and supplementary feeding stations. Urban bird populations are thus more likely to benefit from investment in improving the availability of high quality habitats than controlling squirrel populations or increased investment in supplementary feeding.

  17. Relative Roles of Grey Squirrels, Supplementary Feeding, and Habitat in Shaping Urban Bird Assemblages

    PubMed Central

    Bonnington, Colin; Gaston, Kevin J.; Evans, Karl L.

    2014-01-01

    Non-native species are frequently considered to influence urban assemblages. The grey squirrel Sciurus carolinensis is one such species that is widespread in the UK and is starting to spread across Europe; it predates birds’ nests and can compete with birds for supplementary food. Using distance sampling across the urbanisation intensity gradient in Sheffield (UK) we test whether urban grey squirrels influence avian species richness and density through nest predation and competition for supplementary food sources. We also assess how urban bird assemblages respond to supplementary feeding. We find that grey squirrels slightly reduced the abundance of breeding bird species most sensitive to squirrel nest predation by reducing the beneficial impact of woodland cover. There was no evidence that grey squirrel presence altered relationships between supplementary feeding and avian assemblage structure. This may be because, somewhat surprisingly, supplementary feeding was not associated with the richness or density of wintering bird assemblages. These associations were positive during the summer, supporting advocacy to feed birds during the breeding season and not just winter, but explanatory capacity was limited. The amount of green space and its quality, assessed as canopy cover, had a stronger influence on avian species richness and population size than the presence of grey squirrels and supplementary feeding stations. Urban bird populations are thus more likely to benefit from investment in improving the availability of high quality habitats than controlling squirrel populations or increased investment in supplementary feeding. PMID:25338062

  18. Plant selection for nest building by western lowland gorillas in Cameroon.

    PubMed

    Willie, Jacob; Tagg, Nikki; Petre, Charles-Albert; Pereboom, Zjef; Lens, Luc

    2014-01-01

    We examined 834 nests built by western lowland gorillas in Cameroon between July 2008 and July 2011 to identify the plant species used in their construction. Preference for each plant species for nesting was assessed using a 'preference index' calculated by combining information on the occurrence of each species in the forest and in the nests. Forty-six plant species representing about 15 % of the total number of species in the forest and 26 % of species used for nest building were frequently used by gorillas. Preference levels significantly varied among these species. Nests were mostly built with herbs of the families Marantaceae and Zingiberaceae and woody species such as Manniophyton fulvum (liana) and Alchornea floribunda (shrub). As observed in other gorilla populations, suitability for nest building and availability of gorilla food in stems were the likely determinants of plant selection. The total number of species used per nest ranged from 1 to 11, with an average of 4.9. This is high compared to other sites, emphasizing variability in the availability of nest building materials and habitat differences across the range of the western gorilla. Seasonal changes in the use of different habitat types for nesting did not appear to influence plant use for nest building as there was little variation in plant selection across seasons or the composition of nests. Our findings suggest that gorillas non-randomly select plant species to build nests, and use a particular set of species combined at varying proportions, with no clear seasonal or spatial patterns.

  19. Nesting behavior of the poo-uli

    USGS Publications Warehouse

    Kepler, C.B.; Pratt, T.K.; Ecton, A.M.; Engilis, A.; Fluetsch, K.M.

    1996-01-01

    We describe two sequential nestings of a pair of Poo-uli (Melamprosops phaeosoma), a Hawaiian honeycreeper nearing extinction. Similarities to nesting of most other honeycreepers included: nest site in ohia lehua (Metrosideros polymorpha Gaud.) canopy; breeding in March through June; monogamous breeding system with the putative male helping build the nest, feeding the putative female throughout each nesting event, and feeding the chicks, but not incubating or brooding; and complete nest sanitation. Notable differences were the paucity of songs and calls by the parents and inclusion of snails in the diet of nestlings. Clutch size was probably two eggs for both nests. High winds, rain, or both influenced parental behavior: the female stayed longer on the nest and took shorter recesses in poor weather. Weather did not affect rates at which the male fed the female on the nest; however, the feeding rate increased from the egg to the chick stage probably because food was passed on to the chicks. At nest #2, parents fed young chicks (<14 days old) more often in good than in poor weather; data were insufficient for old chicks. Weather is usually poor throughout the year in the relictual range of the Poo-uli and is likely to impact nesting success. The first nest failed in poor weather. The second fledged a single young 21 days old. Diet of nestlings appeared to consist of a higher proportion of insect larvae than that of older birds, which are reported to eat mostly snails.

  20. Avian nest success in midwestern forests fragmented by agriculture

    USGS Publications Warehouse

    Knutson, M.G.; Niemi, G.J.; Newton, W.E.; Friberg, M.A.

    2004-01-01

    We studied how forest-bird nest success varied by landscape context from 1996 to 1998 in an agricultural region of southeastern Minnesota, southwestern Wisconsin, and northeastern Iowa. Nest success was 48% for all nests, 82% for cavity-nesting species, and 42% for cup-nesting species. Mayfield-adjusted nest success for five common species ranged from 23% for the American Redstart (Setophaga ruticilla) to 43% for the Eastern Wood-Pewee (Contopus virens). Nest success was lowest for open-cup nesters, species that reject Brown-headed Cowbird (Molothrus ater) eggs, species that nest near forest edges, and Neo-tropical migrants. The proportion of forest core area in a 5-km radius around the plot had a weakly negative relationship with daily survival rate of nests for all species pooled and for medium or high canopy nesters, species associated with interior and edge habitats, open-cup nesters, and nests located between 75 and 199 m from an edge. The proportion of forest core area was positively related to daily survival rate only for ground and low nesters. Our findings are in contrast to a number of studies from the eastern United States reporting strong positive associations between forest area and nesting success. Supported models of habitat associations changed with the spatial scale of analysis and included variables not often considered in studies of forest birds, including the proportion of water, shrubs, and grasslands in the landscape. Forest area may not be a strong indicator of nest success in landscapes where all the available forests are fragmented.

  1. Long term productivity of canvasbacks (Aythya valisineria) in a snowpack-driven desert marsh

    USGS Publications Warehouse

    Kruse, Kammie L.; Lovvorn, James R.; Takekawa, John Y.; Mackay, Jeffrey

    2003-01-01

    Ruby Lake, Nevada, is a large palustrine wetland that hosts the southern-most major breeding population of Canvasbacks (Aythya valisineria). That arid marsh, fed by springs derived from mountain snowpack, differs in climate and hydrology from glaciated potholes of the northern prairies where most Canvasbacks breed. Fourteen years of nesting data on Canvasbacks over a 31 year period (1970–2000) were analyzed to determine factors affecting breeding performance at Ruby Lake and whether they differed from those in the prairies. Long-term Mayfield nest success at Ruby Lake (50% of all nests) was in the range of that in the northern prairies (21–65%). Of all Canvasback nests, 73% were parasitized (mostly by Redheads [Aythya americana]) as compared to 83–97% in a large Manitoba marsh and 57–65% in Manitoba potholes. However, as in the northern prairies, nest parasitism generally had little or no effect on either nest success or percentage of host eggs that hatched. In Manitoba potholes, nest success was unrelated to habitat variables measured; but successful nests at Ruby Lake were over shallower water, farther from shore, in wider bands of emergent vegetation, and surrounded by lower stem densities than unsuccessful nests. Water level is the key factor in breeding performance of Canvasbacks at both Ruby Lake and the northern prairies; however, the source of water differs (mountain snowpack at Ruby Lake, direct precipitation in the prairies) and effects of water-level variations are reversed. In small prairie potholes (mostly <0.4 ha) with many mammalian predators, productivity of Canvasbacks (which build floating nests) is increased by high water that floods the emergent fringe. At Ruby Lake, a very large marsh (2,830 ha) with mostly avian predators, Canvasback productivity is decreased by high water that floods interior emergent stands too deeply. Water level at Ruby Lake was highly correlated (multiple R2 = 0.91) with mountain snowpack up to three years earlier, emphasizing the strong effect of climatic variations on wetland birds in that arid region.

  2. The ethological trap: functional and numerical responses of highly efficient invasive predators driving prey extinctions.

    PubMed

    Spencer, Ricky-John; Van Dyke, James U; Thompson, Michael B

    2016-10-01

    Ecological traps are threats to organisms, and exist in a range of biological systems. A subset of ecological trap theory is the "ethological trap," whereby behaviors canalized by past natural selection become traps when environments change rapidly. Invasive predators are major threats to imperiled species and their ability to exploit canalized behaviors of naive prey is particularly important for the establishment of the predator and the decline of the native prey. Our study uses ecological theory to demonstrate that invasive predator controls require shifts in management priorities. Total predation rate (i.e., total response) is the product of both the functional response and numerical response of predators to prey. Functional responses are the changes in the rate of prey consumption by individual predators, relative to prey abundance. Numerical responses are the aggregative rates of prey consumption by all predators relative to prey density, which change with predator density via reproduction or migration, in response to changes in prey density. Traditional invasive predator management methods focus on reducing predator populations, and thus manage for numerical responses. These management efforts fail to manage for functional responses, and may not eliminate impacts of highly efficient individual predators. We explore this problem by modeling the impacts of functional and numerical responses of invasive foxes depredating imperiled Australian turtle nests. Foxes exhibit exceptionally efficient functional responses. A single fox can destroy >95% of turtle nests in a nesting area, which eliminates juvenile recruitment. In this case, the ethological trap is the "Arribada" nesting strategy, an emergent behavior whereby most turtles in a population nest simultaneously in the same nesting grounds. Our models show that Arribada nesting events do not oversaturate foxes, and small numbers of foxes depredate all of the nests in a given Arribada. Widely scattering nests may reduce fox predation rates, but the long generation times of turtles combined with their rapid recent decline suggests that evolutionary responses in nesting strategy may be unlikely. Our study demonstrates that reducing populations of highly efficient invasive predators is insufficient for preserving native prey species. Instead, management must reduce individual predator efficiency, independent of reducing predator population size. © 2016 by the Ecological Society of America.

  3. Methods for measuring populations of arboreal rodents.

    Treesearch

    Andrew B. Carey; Brian L. Biswell; Joseph W. Witt

    1991-01-01

    Three arboreal rodents are sensitive indicators of forest ecosystem function in the Pacific Northwest. The northern flying squirrel (Glaucomys sabrinus) is mycophagous, cavity-nesting, and a major prey of the spotted owl (Strix occidentalis). The red tree vole (Phenacomys longicaudus) is restricted to trees...

  4. Florida Harvester Ant Nest Architecture, Nest Relocation and Soil Carbon Dioxide Gradients

    PubMed Central

    Tschinkel, Walter R.

    2013-01-01

    Colonies of the Florida harvester ant, Pogonomyrmex badius, excavate species-typical subterranean nests up the 3 m deep with characteristic vertical distribution of chamber area/shape, spacing between levels and vertical arrangement of the ants by age and brood stage. Colonies excavate and occupy a new nest about once a year, and doing so requires that they have information about the depth below ground. Careful excavation and mapping of vacated and new nests revealed that there was no significant difference between the old and new nests in any measure of nest size, shape or arrangement. Colonies essentially built a replicate of the just-vacated nest (although details differed), and they did so in less than a week. The reason for nest relocation is not apparent. Tschinkel noted that the vertical distribution of chamber area, worker age and brood type was strongly correlated to the soil carbon dioxide gradient, and proposed that this gradient serves as a template for nest excavation and vertical distribution. To test this hypothesis, the carbon dioxide gradient of colonies that were just beginning to excavate a new nest was eliminated by boring 6 vent holes around the forming nest, allowing the soil CO2 to diffuse into the atmosphere and eliminating the gradient. Sadly, neither the nest architecture nor the vertical ant distribution of vented nests differed from either unvented control or from their own vacated nest. In a stronger test, workers excavated a new nest under a reversed carbon dioxide gradient (high concentration near the surface, low below). Even under these conditions, the new and old nests did not differ significantly, showing that the soil carbon dioxide gradient does not serve as a template for nest construction or vertical worker distribution. The possible importance of soil CO2 gradients for soil-dwelling animals is discussed. PMID:23555829

  5. Eggshell Porosity Provides Insight on Evolution of Nesting in Dinosaurs.

    PubMed

    Tanaka, Kohei; Zelenitsky, Darla K; Therrien, François

    2015-01-01

    Knowledge about the types of nests built by dinosaurs can provide insight into the evolution of nesting and reproductive behaviors among archosaurs. However, the low preservation potential of their nesting materials and nesting structures means that most information can only be gleaned indirectly through comparison with extant archosaurs. Two general nest types are recognized among living archosaurs: 1) covered nests, in which eggs are incubated while fully covered by nesting material (as in crocodylians and megapodes), and 2) open nests, in which eggs are exposed in the nest and brooded (as in most birds). Previously, dinosaur nest types had been inferred by estimating the water vapor conductance (i.e., diffusive capacity) of their eggs, based on the premise that high conductance corresponds to covered nests and low conductance to open nests. However, a lack of statistical rigor and inconsistencies in this method render its application problematic and its validity questionable. As an alternative we propose a statistically rigorous approach to infer nest type based on large datasets of eggshell porosity and egg mass compiled for over 120 extant archosaur species and 29 archosaur extinct taxa/ootaxa. The presence of a strong correlation between eggshell porosity and nest type among extant archosaurs indicates that eggshell porosity can be used as a proxy for nest type, and thus discriminant analyses can help predict nest type in extinct taxa. Our results suggest that: 1) covered nests are likely the primitive condition for dinosaurs (and probably archosaurs), and 2) open nests first evolved among non-avian theropods more derived than Lourinhanosaurus and were likely widespread in non-avian maniraptorans, well before the appearance of birds. Although taphonomic evidence suggests that basal open nesters (i.e., oviraptorosaurs and troodontids) were potentially the first dinosaurs to brood their clutches, they still partially buried their eggs in sediment. Open nests with fully exposed eggs only became widespread among Euornithes. A potential co-evolution of open nests and brooding behavior among maniraptorans may have freed theropods from the ground-based restrictions inherent to covered nests and allowed the exploitation of alternate nesting locations. These changes in nesting styles and behaviors thus may have played a role in the evolutionary success of maniraptorans (including birds).

  6. Eggshell Porosity Provides Insight on Evolution of Nesting in Dinosaurs

    PubMed Central

    2015-01-01

    Knowledge about the types of nests built by dinosaurs can provide insight into the evolution of nesting and reproductive behaviors among archosaurs. However, the low preservation potential of their nesting materials and nesting structures means that most information can only be gleaned indirectly through comparison with extant archosaurs. Two general nest types are recognized among living archosaurs: 1) covered nests, in which eggs are incubated while fully covered by nesting material (as in crocodylians and megapodes), and 2) open nests, in which eggs are exposed in the nest and brooded (as in most birds). Previously, dinosaur nest types had been inferred by estimating the water vapor conductance (i.e., diffusive capacity) of their eggs, based on the premise that high conductance corresponds to covered nests and low conductance to open nests. However, a lack of statistical rigor and inconsistencies in this method render its application problematic and its validity questionable. As an alternative we propose a statistically rigorous approach to infer nest type based on large datasets of eggshell porosity and egg mass compiled for over 120 extant archosaur species and 29 archosaur extinct taxa/ootaxa. The presence of a strong correlation between eggshell porosity and nest type among extant archosaurs indicates that eggshell porosity can be used as a proxy for nest type, and thus discriminant analyses can help predict nest type in extinct taxa. Our results suggest that: 1) covered nests are likely the primitive condition for dinosaurs (and probably archosaurs), and 2) open nests first evolved among non-avian theropods more derived than Lourinhanosaurus and were likely widespread in non-avian maniraptorans, well before the appearance of birds. Although taphonomic evidence suggests that basal open nesters (i.e., oviraptorosaurs and troodontids) were potentially the first dinosaurs to brood their clutches, they still partially buried their eggs in sediment. Open nests with fully exposed eggs only became widespread among Euornithes. A potential co-evolution of open nests and brooding behavior among maniraptorans may have freed theropods from the ground-based restrictions inherent to covered nests and allowed the exploitation of alternate nesting locations. These changes in nesting styles and behaviors thus may have played a role in the evolutionary success of maniraptorans (including birds). PMID:26605799

  7. An 'ecological trap' for yellow warbler nest microhabitat selection

    Treesearch

    Quresh S. Latif; Sacha K. Heath; John T. Rotenberry

    2011-01-01

    Contrary to assumptions of habitat selection theory, fi eld studies frequently detect ‘ ecological traps ’ , where animals prefer habitats conferring lower fi tness than available alternatives. Evidence for traps includes cases where birds prefer breeding habitats associated with relatively high nest predation rates despite the importance of nest survival to avian fi...

  8. Sex ratio estimations of loggerhead sea turtle hatchlings at Kuriat islands, Tunisia: can minor nesting sites contribute to compensate globally female-biased sex ratio?

    PubMed

    Jribi, Imed; Bradai, Mohamed Nejmeddine

    2014-01-01

    Hatchling sex ratios in the loggerhead turtle Caretta caretta were estimated by placing electronic temperature recorders in seven nests at Kuriat islands (Tunisia) during the 2013 nesting season. Based on the mean temperatures during the middle third of the incubation period, and on incubation duration, the sex ratio of hatchlings at Kuriat islands was highly male-biased. Presently, the majority of hatchling sex ratio studies are focused on major nesting areas, whereby the sex ratios are universally believed to be heavily female-biased. Here we present findings from a minor nesting site in the Mediterranean, where the hatchling sex ratio was found to be male-biased, suggesting a potential difference between major and minor nesting sites.

  9. High, low, or familiar? Nest site preferences of experienced laying hens.

    PubMed

    Krause, E T; Schrader, L

    2018-05-22

    1. The aim of this study was to investigate which nest heights are preferred by laying hens in the absence of familiar nest locations and whether preferred nest heights are more attractive than a familiar location. In two experiments, a total of 108 hens of four different layer breeds, which were at least 50 weeks of age, were studied. 2. In the first experiment, hens were given individual free choice between nests for 1-week at four different heights (0 cm, 39 cm, 78 cm, and 117 cm above ground). Hens of the four breeds differed in their nest height preferences (P = 0.0013). However, hens of three breeds preferred ground level nests (P < 0.007) and the fourth line showed an equal preference for the ground level and level three, the latter level corresponding to the height of the nests in their home compartments. 4. In the second experiment, hens from the four breeds were given a choice between ground level nests and nests at a familiar location, i.e. at the same location as in their home compartment. Hens of all strains preferred the familiar nest location (P = 0.002) and preferences did not differ between strains (P = 0.77). 5. Laying hens seem to prefer nests at ground level in the absence of a familiar nest. However, if possible, experienced 50 week old hens continue to use a familiar nest location instead of a ground nest location. The results are discussed with respect to a potential primary preference that may be modifiable by experience and with respect to possible relevance in commercial housing.

  10. Landscape-moderated bird nest predation in hedges and forest edges

    NASA Astrophysics Data System (ADS)

    Ludwig, Martin; Schlinkert, Hella; Holzschuh, Andrea; Fischer, Christina; Scherber, Christoph; Trnka, Alfréd; Tscharntke, Teja; Batáry, Péter

    2012-11-01

    Landscape-scale agricultural intensification has caused severe declines in biodiversity. Hedges and forest remnants may mitigate biodiversity loss by enhancing landscape heterogeneity and providing habitat to a wide range of species, including birds. However, nest predation, the major cause of reproductive failure of birds, has been shown to be higher in forest edges than in forest interiors. Little is known about how spatial arrangement (configuration) of hedges affects the avian nest predation. We performed an experiment with artificial ground and elevated nests (resembling yellowhammer and whitethroat nests) baited with quail and plasticine eggs. Nests were placed in three habitat types with different degrees of isolation from forests: forest edges, hedges connected to forests and hedges isolated from forests. Nest predation was highest in forest edges, lowest in hedges connected to forests and intermediate in isolated hedges. In the early breeding season, we found similar nest predation on ground and elevated nests, but in the late breeding season nest predation was higher on ground nests than on elevated nests. Small mammals were the main predators of ground nests and appeared to be responsible for the increase in predation from early to late breeding season, whereas the elevated nests were mainly depredated by small birds and small mammals. High predation pressure at forest edges was probably caused by both forest and open-landscape predators. The influence of forest predators may be lower at hedges, leading to lower predation pressure than in forest edges. Higher predation pressure in isolated than connected hedges might be an effect of concentration of predators in these isolated habitats. We conclude that landscape configuration of hedges is important in nest predation, with connected hedges allowing higher survival than isolated hedges and forest edges.

  11. Single molecule quantitation and sequencing of rare translocations using microfluidic nested digital PCR.

    PubMed

    Shuga, Joe; Zeng, Yong; Novak, Richard; Lan, Qing; Tang, Xiaojiang; Rothman, Nathaniel; Vermeulen, Roel; Li, Laiyu; Hubbard, Alan; Zhang, Luoping; Mathies, Richard A; Smith, Martyn T

    2013-09-01

    Cancers are heterogeneous and genetically unstable. New methods are needed that provide the sensitivity and specificity to query single cells at the genetic loci that drive cancer progression, thereby enabling researchers to study the progression of individual tumors. Here, we report the development and application of a bead-based hemi-nested microfluidic droplet digital PCR (dPCR) technology to achieve 'quantitative' measurement and single-molecule sequencing of somatically acquired carcinogenic translocations at extremely low levels (<10(-6)) in healthy subjects. We use this technique in our healthy study population to determine the overall concentration of the t(14;18) translocation, which is strongly associated with follicular lymphoma. The nested dPCR approach improves the detection limit to 1×10(-7) or lower while maintaining the analysis efficiency and specificity. Further, the bead-based dPCR enabled us to isolate and quantify the relative amounts of the various clonal forms of t(14;18) translocation in these subjects, and the single-molecule sensitivity and resolution of dPCR led to the discovery of new clonal forms of t(14;18) that were otherwise masked by the conventional quantitative PCR measurements. In this manner, we created a quantitative map for this carcinogenic mutation in this healthy population and identified the positions on chromosomes 14 and 18 where the vast majority of these t(14;18) events occur.

  12. Development of a reverse transcription polymerase chain reaction method for yellow fever virus detection.

    PubMed

    Méndez, María C; Domingo, Cristina; Tenorio, Antonio; Pardo, Lissethe C; Rey, Gloria J; Méndez, Jairo A

    2013-09-01

    Yellow fever is considered a re-emerging disease and is endemic in tropical regions of Africa and South America. At present, there are no standardized or commercialized kits available for yellow fever virus detection. Therefore, diagnosis must be made by time-consuming routine techniques, and sometimes, the virus or its proteins are not detected. Furthermore, co-circulation with other flaviviruses, including dengue virus, increases the difficulty of diagnosis. To develop a specific reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR-based assay to improve the detection and diagnosis of yellow fever virus using both serum and fresh tissue samples. RT-PCR primers were designed to amplify a short fragment of all yellow fever virus genotypes reported. A second set of primers was used in a nested PCR to increase sensitivity. Thirty-three clinical samples were tested with the standardized reaction. The expected amplicon was obtained in 25 out of 33 samples analyzed using this approach, and 2 more samples tested positive after a subsequent nested PCR approach. This improved technique not only ensures the specific detection of a wide range of yellow fever virus genotypes but also may increase the sensitivity of detection by introducing a second round of amplification, allowing a rapid differential diagnosis between dengue and yellow fever infection, which is required for effective surveillance and opportune epidemiologic measures.

  13. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Belinsky, Steven A; Palmisano, William A

    A molecular marker-based method for monitoring and detecting cancer in humans. Aberrant methylation of gene promoters is a marker for cancer risk in humans. A two-stage, or "nested" polymerase chain reaction method is disclosed for detecting methylated DNA sequences at sufficiently high levels of sensitivity to permit cancer screening in biological fluid samples, such as sputum, obtained non-invasively. The method is for detecting the aberrant methylation of the p16 gene, O 6-methylguanine-DNA methyltransferase gene, Death-associated protein kinase gene, RAS-associated family 1 gene, or other gene promoters. The method offers a potentially powerful approach to population-based screening for the detection ofmore » lung and other cancers.« less

  14. Optimization for minimum sensitivity to uncertain parameters

    NASA Technical Reports Server (NTRS)

    Pritchard, Jocelyn I.; Adelman, Howard M.; Sobieszczanski-Sobieski, Jaroslaw

    1994-01-01

    A procedure to design a structure for minimum sensitivity to uncertainties in problem parameters is described. The approach is to minimize directly the sensitivity derivatives of the optimum design with respect to fixed design parameters using a nested optimization procedure. The procedure is demonstrated for the design of a bimetallic beam for minimum weight with insensitivity to uncertainties in structural properties. The beam is modeled with finite elements based on two dimensional beam analysis. A sequential quadratic programming procedure used as the optimizer supplies the Lagrange multipliers that are used to calculate the optimum sensitivity derivatives. The method was perceived to be successful from comparisons of the optimization results with parametric studies.

  15. Tropical flatback turtle (Natator depressus) embryos are resilient to the heat of climate change.

    PubMed

    Howard, Robert; Bell, Ian; Pike, David A

    2015-10-01

    Climate change is threatening reproduction of many ectotherms by increasing nest temperatures, potentially making it more difficult for females to locate nest sites that provide suitable incubation regimes during embryonic development. Elevated nest temperatures could increase the incidence of embryonic mortality and/or maladaptive phenotypes. We investigated whether elevated nest temperatures reduce hatching success in tropical flatback turtles (Natator depressus) nesting in the Gulf of Carpentaria, Australia. Egg incubation treatments began at 29.5°C and progressively increased in temperature throughout incubation, up to maxima of 31, 32, 33, 34 and 35°C. Elevated nest temperatures did not reduce hatching success or hatchling body size relative to control temperatures (29.5°C), but did speed up embryonic development. A combination of sudden exposure to high temperatures during the first 2 weeks of incubation (>36°C for 48 h) and prolonged warming throughout incubation (from 29.5-35°C) did not reduce hatching success. We also recorded an unusually high pivotal sex-determining temperature in this flatback turtle population relative to other sea turtle populations: an equal ratio of male and female hatchlings is produced at ∼30.4°C. This adaptation may allow some flatback turtle populations to continue producing large numbers of hatchlings of both sexes under the most extreme climate change scenarios. Some tropical populations of nesting flatbacks may possess important adaptations to high-temperature incubation environments, which are not found in more southerly temperate populations. © 2015. Published by The Company of Biologists Ltd.

  16. Greater sage-grouse nest predators in the Virginia Mountains of northwestern Nevada

    USGS Publications Warehouse

    Lockyer, Zachary B.; Coates, Peter S.; Casazza, Michael L.; Espinosa, Shawn; Delehanty, David J.

    2013-01-01

    Greater sage-grouse Centrocercus urophasianus, hereafter sage-grouse, populations have declined across their range due to the loss, degradation, and fragmentation of habitat. Habitat alterations can lead not only to vegetative changes but also to shifts in animal behavior and predator composition that may influence population vital rates, such as nest success. For example, common ravens Corvus corax are sage-grouse nest predators, and common raven abundance is positively associated with human-caused habitat alterations. Because nest success is a central component to sage-grouse population persistence, research that identifies factors influencing nest success will better inform conservation efforts. We used videography to unequivocally identify sage-grouse nest predators within the Virginia Mountains of northwestern Nevada, USA, from 2009 to 2011 and used maximum likelihood to calculate daily probability of nest survival. In the Virginia Mountains, fires, energy exploration, and other anthropogenic activities have altered historic sage-grouse habitat. We monitored 71 sage-grouse nests during the study, placing video cameras at 39 nests. Cumulative nest survival for all nests was 22.4% (95% CI, 13.0–33.4%), a survival rate that was significantly lower than other published results for sage-grouse in the Great Basin. Depredation was the primary cause for nest failure in our study (82.5%), and common ravens were the most frequent sage-grouse nest predator, accounting for 46.7% of nest depredations. We also successfully documented a suite of mammalian and reptilian species depredating sage-grouse nests, including some predators never previously confirmed in the literature to be sage-grouse nest predators (i.e., bobcats Lynx rufus and long-tailed weasels Mephitis frenata). Within the high elevation, disturbed habitat of the Virginia Mountains, low sage-grouse nest success may be limiting sage-grouse population growth. These results suggest that management actions that restore habitat in the Virginia Mountains and decrease anthropogenic subsidies of ravens will benefit sage-grouse.

  17. Development of high-resolution multi-scale modelling system for simulation of coastal-fluvial urban flooding

    NASA Astrophysics Data System (ADS)

    Comer, Joanne; Indiana Olbert, Agnieszka; Nash, Stephen; Hartnett, Michael

    2017-02-01

    Urban developments in coastal zones are often exposed to natural hazards such as flooding. In this research, a state-of-the-art, multi-scale nested flood (MSN_Flood) model is applied to simulate complex coastal-fluvial urban flooding due to combined effects of tides, surges and river discharges. Cork city on Ireland's southwest coast is a study case. The flood modelling system comprises a cascade of four dynamically linked models that resolve the hydrodynamics of Cork Harbour and/or its sub-region at four scales: 90, 30, 6 and 2 m. Results demonstrate that the internalization of the nested boundary through the use of ghost cells combined with a tailored adaptive interpolation technique creates a highly dynamic moving boundary that permits flooding and drying of the nested boundary. This novel feature of MSN_Flood provides a high degree of choice regarding the location of the boundaries to the nested domain and therefore flexibility in model application. The nested MSN_Flood model through dynamic downscaling facilitates significant improvements in accuracy of model output without incurring the computational expense of high spatial resolution over the entire model domain. The urban flood model provides full characteristics of water levels and flow regimes necessary for flood hazard identification and flood risk assessment.

  18. Size determines antennal sensitivity and behavioral threshold to odors in bumblebee workers

    NASA Astrophysics Data System (ADS)

    Spaethe, Johannes; Brockmann, Axel; Halbig, Christine; Tautz, Jürgen

    2007-09-01

    The eusocial bumblebees exhibit pronounced size variation among workers of the same colony. Differently sized workers engage in different tasks (alloethism); large individuals are found to have a higher probability to leave the colony and search for food, whereas small workers tend to stay inside the nest and attend to nest duties. We investigated the effect of size variation on morphology and physiology of the peripheral olfactory system and the behavioral response thresholds to odors in workers of Bombus terrestris. Number and density of olfactory sensilla on the antennae correlate significantly with worker size. Consistent with these morphological changes, we found that antennal sensitivity to odors increases with body size. Antennae of large individuals show higher electroantennogram responses to a given odor concentration than those of smaller nestmates. This finding indicates that large antennae exhibit an increased capability to catch odor molecules and thus are more sensitive to odors than small antennae. We confirmed this prediction in a dual choice behavioral experiment showing that large workers indeed are able to respond correctly to much lower odor concentrations than small workers. Learning performance in these experiments did not differ between small and large bumblebees. Our results clearly show that, in the social bumblebees, variation in olfactory sensilla number due to size differences among workers strongly affects individual odor sensitivity. We speculate that superior odor sensitivity of large workers has favored size-related division of labor in bumblebee colonies.

  19. Magnetoreception in Eusocial Insects: An Update

    USDA-ARS?s Scientific Manuscript database

    Behavioral experiments for magnetoreception in eusocial insects in the last decade are reviewed. Ants and bees use the geomagnetic field to orient and navigate in areas around their nests and in migratory paths. Bees show sensitivity to small changes in magnetic fields in conditioning experiments a...

  20. Diagnosis of Gestational, Congenital, and Placental Malaria in Colombia: Comparison of the Efficacy of Microscopy, Nested Polymerase Chain Reaction, and Histopathology

    PubMed Central

    Campos, Ivón M.; Uribe, Mary L.; Cuesta, Carolina; Franco-Gallego, Alexander; Carmona-Fonseca, Jaime; Maestre, Amanda

    2011-01-01

    The technical capability of different methods to diagnose Plasmodium in maternal peripheral blood, placenta, and umbilical cord blood has not been assessed in Colombia and seldom explored in other malaria-endemic regions. We designed a study to compare the technical and the operational-economical performances of light microscopy (LM), nested polymerase chain reaction (nPCR), and histopathology (HP). In maternal blood, LM had 41% sensitivity and 100% specificity and in placental blood, 35% and 100%, respectively, compared with nPCR. In placental tissue, LM had 33% sensitivity and 95% specificity; and nPCR 47% and 77%, respectively; compared with HP. Light microscopy had the best operational-economical qualification. We concluded that nPCR and HP performed better compared with LM, but field implementation of these two techniques remains a problem. Therefore, LM is recommended as the gold standard for diagnosis of gestational malaria and placental blood infection in the field. PMID:21633030

  1. Comparison between conventional and molecular methods for diagnosis of bovine babesiosis (Babesia bovis infection) in tick infested cattle in upper Egypt.

    PubMed

    Al-Hosary, Amira A T

    2017-03-01

    Ticks and tick-borne diseases are the main problems affecting the livestock production in Egypt. Bovine babesiosis has adverse effects on the animal health and production. A comparison of Giemsa stained blood smears, polymerase chain reaction (PCR) and nested PCR (nPCR) assays for detection of Babesia bovis infection in Egyptian Baladi cattle ( Bos taurus ) in reference to reverse line blot was carried out. The sensitivity of PCR and nested PCR (nPCR) assays were 65 and 100 % respectively. Giemsa stained blood smears showed the lowest sensitivity (30 %). According to these results using of PCR and nPCR target for B. bovis , [BBOV-IV005650 (BV5650)] gene are suitable for diagnosis of B. bovis infection. The 18Ss rRNA partial sequence confirmed that all the positive samples were Babesia bovis and all of them were deposited in the GenBank databases (Accession No: KM455548, KM455549 and KM455550).

  2. Modified use of methylene blue in the tissue compression technique to detect sarcocysts in meat-producing animals.

    PubMed

    Ng, Yit Han; Subramaniam, Vellayan; Lau, Yee Ling

    2015-11-30

    Sarcocystosis in meat-producing animals is a major cause of reduced productivity in many countries, especially those that rely on agriculture. Although several diagnostic methods are available to detect sarcocystosis, many are too time-consuming for routine use in abattoirs and meat inspection centers, where large numbers of samples need to be tested. This study aimed to compare the sensitivity of the methylene blue tissue preparation, unstained tissue preparation and nested PCR in the detection of sarcocysts in tissue samples. Approximately three-fold more sarcocysts were detected in methylene blue-stained tissue compared to unstained controls (McNemar's test: P<0.01). Test sensitivity was comparable to that of the gold standard for sarcocyst detection, nested polymerase chain reaction. These results suggest that methylene blue can be used in tissue compression as a rapid, safe, and inexpensive technique for the detection of ruminant sarcocystosis in abattoirs. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. High-temperature, high-pressure bonding of nested tubular metallic components

    DOEpatents

    Quinby, T.C.

    A tool is described for effecting high-temperature, high-compression bonding between the confronting faces of nested, tubular, metallic components. In a typical application, the tool is used to produce tubular target assemblies for irradiation in nuclear reactors or particle accelerators. The target assembly comprising a uranum foil and an aluninum-alloy substrate. The tool is composed of graphite. It comprises a tubular restraining member in which a mechanically expandable tubular core is mounted to form an annulus. The components to be bonded are mounted in nested relation in the annulus. The expandable core is formed of individually movable, axially elongated segments whose outer faces cooperatively define a cylindrical pressing surface and whose inner faces cooperatively define two opposed, inwardly tapered, axial bores. Tapered rams extend into the bores. The loaded tool is mounted in a conventional hot-press provided with evacuation means, heaters for maintaining its interior at bonding temperature, and hydraulic cylinders for maintaining a selected inwardly directed pressure on the tapered rams. With the hot-press evacuated and the loaded tool at the desired temperature, the cylinders are actuated to apply the selected pressure to the rams. The rams in turn expand the segmented core to maintain the nested components in compression against the restraining member. These conditions are maintained until the confronting faces of the nested components are joined in a continuous, uniform bond characterized by high thermal conductivity.

  4. Local habitat and landscape influence predation of bird nests on afforested Mediterranean cropland

    NASA Astrophysics Data System (ADS)

    Sánchez-Oliver, J. S.; Rey Benayas, J. M.; Carrascal, L. M.

    2014-07-01

    Afforestation programs such as the one promoted by the EU Common Agrarian Policy have contributed to spread tree plantations on former cropland. Nevertheless these afforestations may cause severe damage to open habitat species, especially birds of high conservation value. We investigated predation of artificial bird nests at young tree plantations and at the open farmland habitat adjacent to the tree plantations in central Spain. Predation rates were very high at both tree plantations (95.6%) and open farmland habitat (94.2%) after two and three week exposure. Plantation edge/area ratio and development of the tree canopy decreased predation rates and plantation area and magpie (Pica pica) abundance increased predation rates within tree plantations, which were also affected by land use types around plantations. The area of nearby tree plantations (positive effect), distance to the tree plantation edge (negative effect), and habitat type (mainly attributable to the location of nests in vineyards) explained predation rates at open farmland habitat. We conclude that predation rates on artificial nests were particularly high and rapid at or nearby large plantations, with high numbers of magpies and low tree development, and located in homogenous landscapes dominated by herbaceous crops and pastures with no remnants of semi-natural woody vegetation. Landscape planning should not favour tree plantations as the ones studied here in Mediterranean agricultural areas that are highly valuable for ground-nesting bird species.

  5. Retirement investment theory explains patterns in songbird nest-site choice

    USGS Publications Warehouse

    Streby, Henry M.; Refsnider, Jeanine M.; Peterson, Sean M.; Andersen, David E.

    2014-01-01

    When opposing evolutionary selection pressures act on a behavioural trait, the result is often stabilizing selection for an intermediate optimal phenotype, with deviations from the predicted optimum attributed to tracking a moving target, development of behavioural syndromes or shifts in riskiness over an individual's lifetime. We investigated nest-site choice by female golden-winged warblers, and the selection pressures acting on that choice by two fitness components, nest success and fledgling survival. We observed strong and consistent opposing selection pressures on nest-site choice for maximizing these two fitness components, and an abrupt, within-season switch in the fitness component birds prioritize via nest-site choice, dependent on the time remaining for additional nesting attempts. We found that females consistently deviated from the predicted optimal behaviour when choosing nest sites because they can make multiple attempts at one fitness component, nest success, but only one attempt at the subsequent component, fledgling survival. Our results demonstrate a unique natural strategy for balancing opposing selection pressures to maximize total fitness. This time-dependent switch from high to low risk tolerance in nest-site choice maximizes songbird fitness in the same way a well-timed switch in human investor risk tolerance can maximize one's nest egg at retirement. Our results also provide strong evidence for the adaptive nature of songbird nest-site choice, which we suggest has been elusive primarily due to a lack of consideration for fledgling survival.

  6. Inheritance of nesting behaviour across natural environmental variation in a turtle with temperature-dependent sex determination.

    PubMed

    McGaugh, Suzanne E; Schwanz, Lisa E; Bowden, Rachel M; Gonzalez, Julie E; Janzen, Fredric J

    2010-04-22

    Nesting behaviour is critical for reproductive success in oviparous organisms with no parental care. In organisms where sex is determined by incubation temperature, nesting behaviour may be a prime target of selection in response to unbalanced sex ratios. To produce an evolutionary change in response to sex-ratio selection, components of nesting behaviour must be heritable. We estimated the field heritability of two key components of nesting behaviour in a population of painted turtles (Chrysemys picta) with temperature-dependent sex determination by applying the 'animal model' to a pedigree reconstructed from genotype data. We obtained estimates of low to non-detectable heritability using repeated records across all environments. We then determined environment-specific heritability by grouping records with similar temperatures for the winter preceding the nesting season, a variable known to be highly associated with our two traits of interest, nest vegetation cover and Julian date of nesting. The heritability estimates of nest vegetation cover and Julian date of nesting were qualitatively highest and significant, or nearly so, after hot winters. Additive genetic variance for these traits was not detectable after cold winters. Our analysis suggests that the potential for evolutionary change of nesting behaviour may be dependent on the thermal conditions of the preceding winter, a season that is predicted to be especially subject to climate change.

  7. FVCOM one-way and two-way nesting using ESMF: Development and validation

    NASA Astrophysics Data System (ADS)

    Qi, Jianhua; Chen, Changsheng; Beardsley, Robert C.

    2018-04-01

    Built on the Earth System Modeling Framework (ESMF), the one-way and two-way nesting methods were implemented into the unstructured-grid Finite-Volume Community Ocean Model (FVCOM). These methods help utilize the unstructured-grid multi-domain nesting of FVCOM with an aim at resolving the multi-scale physical and ecosystem processes. A detail of procedures on implementing FVCOM into ESMF was described. The experiments were made to validate and evaluate the performance of the nested-grid FVCOM system. The first was made for a wave-current interaction case with a two-domain nesting with an emphasis on qualifying a critical need of nesting to resolve a high-resolution feature near the coast and harbor with little loss in computational efficiency. The second was conducted for the pseudo river plume cases to examine the differences in the model-simulated salinity between one-way and two-way nesting approaches and evaluate the performance of mass conservative two-way nesting method. The third was carried out for the river plume case in the realistic geometric domain in Mass Bay, supporting the importance for having the two-way nesting for coastal-estuarine integrated modeling. The nesting method described in this paper has been used in the Northeast Coastal Ocean Forecast System (NECOFS)-a global-regional-coastal nesting FVCOM system that has been placed into the end-to-end forecast and hindcast operations since 2007.

  8. Female turtles from hot nests: is it duration of incubation or proportion of development at high temperatures that matters?

    PubMed

    Georges, Arthur

    1989-11-01

    Mean daily temperature in natural nests of freshwater turtles with temperature-dependent sex determination is known to be a poor predictor of hatchling sex ratios when nest temperatures fluctuate. To account for this, a model was developed on the assumption that females will emerge from eggs when more than half of embryonic development occurs above the threshold temperature for sex determination rather than from eggs that spend more than half their time above the threshold. The model is consistent with previously published data and in particular explains the phenomenon whereby the mean temperature that best distinguishes between male and female nests decreases with increasing variability in nest temperature. The model, if verified by controlled experiments, has important implications for our understanding of temperature-dependent sex determination in natural nests. Both mean nest temperature and "hours spent above the threshold" will be poor predictors of hatchling sex ratios. Studies designed to investigate latitudinal trends and inter-specific differences in the threshold temperature will need to consider latitudinal and inter-specific variation in the magnitude of diel fluctuations in nest temperature, and variation in factors influencing the magnitude of those fluctuations, such as nest depth. Furthermore, any factor that modifies the relationship between developmental rate and temperature can be expected to influence hatchling sex ratios in natural nests, especially when nest temperatures are close to the threshold.

  9. Post-Mating Sexual Behaviors of Oriental Storks (Ciconia boyciana) in Captivity.

    PubMed

    Yoon, Jongmin; Ha, Hae-Sook; Jung, Jung-Shim; Park, Shi-Ryong

    2015-08-01

    Parental behaviors that vary by sex and breeding stage facilitate the survival of offspring in birds. Females invest in reproducing according to the level of direct and/or indirect benefits provided by males. Males face trade-offs from defending territories or nests, providing for the young, and seeking additional mating opportunities. We examined whether post-mating sexual behaviors such as courtship display and nest-building were associated with reproductive investments made by males and females in the current study. The oriental stork (Ciconia boyciana) is known as a solitary breeder that nests in tall trees and provides biparental care with long post-hatching development. We filmed parental behaviors in seven stork pairs in captivity for 655 hours (245 hours during nest-building, 218 hours during incubation, 328 hours during nestling, and 192 hours during the fledgling period). We found that paired mating behaviors such as synchronous allopreening and bill-clattering were highly skewed to the pre-incubation period. Males participated in nest-building more than females although both sexes shared similar parental care during the incubation and nestling period. Male nest-building was negatively correlated with male nest attentiveness only during the nestling period. Our results suggest that male oriental storks with higher nest-building effort might spend more time taking precautions against conspecific intruders or nest predators near the nests with chicks while the females were attending the nests in the form of biparental care.

  10. Strong selection on mandible and nest features in a carpenter bee that nests in two sympatric host plants

    PubMed Central

    Flores-Prado, Luis; Pinto, Carlos F; Rojas, Alejandra; Fontúrbel, Francisco E

    2014-01-01

    Host plants are used by herbivorous insects as feeding or nesting resources. In wood-boring insects, host plants features may impose selective forces leading to phenotypic differentiation on traits related to nest construction. Carpenter bees build their nests in dead stems or dry twigs of shrubs and trees; thus, mandibles are essential for the nesting process, and the nest is required for egg laying and offspring survival. We explored the shape and intensity of natural selection on phenotypic variation on three size measures of the bees (intertegular width, wing length, and mandible area) and two nest architecture measures (tunnel length and diameter) on bees using the native species Chusquea quila (Poaceae), and the alloctonous species Rubus ulmifolius (Rosaceae), in central Chile. Our results showed significant and positive linear selection gradients for tunnel length on both hosts, indicating that bees building long nests have more offspring. Bees with broader mandibles show greater fitness on C. quila but not on R. ulmifolius. Considering that C. quila represents a selective force on mandible area, we hypothesized a high adaptive value of this trait, resulting in higher fitness values when nesting on this host, despite its wood is denser and hence more difficult to be bored. PMID:24963379

  11. Strong selection on mandible and nest features in a carpenter bee that nests in two sympatric host plants.

    PubMed

    Flores-Prado, Luis; Pinto, Carlos F; Rojas, Alejandra; Fontúrbel, Francisco E

    2014-05-01

    Host plants are used by herbivorous insects as feeding or nesting resources. In wood-boring insects, host plants features may impose selective forces leading to phenotypic differentiation on traits related to nest construction. Carpenter bees build their nests in dead stems or dry twigs of shrubs and trees; thus, mandibles are essential for the nesting process, and the nest is required for egg laying and offspring survival. We explored the shape and intensity of natural selection on phenotypic variation on three size measures of the bees (intertegular width, wing length, and mandible area) and two nest architecture measures (tunnel length and diameter) on bees using the native species Chusquea quila (Poaceae), and the alloctonous species Rubus ulmifolius (Rosaceae), in central Chile. Our results showed significant and positive linear selection gradients for tunnel length on both hosts, indicating that bees building long nests have more offspring. Bees with broader mandibles show greater fitness on C. quila but not on R. ulmifolius. Considering that C. quila represents a selective force on mandible area, we hypothesized a high adaptive value of this trait, resulting in higher fitness values when nesting on this host, despite its wood is denser and hence more difficult to be bored.

  12. Superpredator proximity and landscape characteristics alters nest site selection and breeding success of a subordinate predator.

    PubMed

    Atuo, Fidelis Akunke; O'Connell, Timothy John

    2018-03-01

    Selecting nesting habitat that minimizes predation risk but maximizes foraging success is one of the most important decisions in avian life history. This takes on added complexity when a predator is faced with the challenge of avoiding fellow predators. We assessed the importance of local and landscape vegetation, food abundance, and predation risk on nest site selection and nest survival in a subordinate raptor (Mississippi Kite; Ictinia mississippiensis) nesting in proximity to two superpredators, Red-tailed hawk (Buteo jamaicensis) and Great horned owl (Bubo virginianus). All three species nested in trees in a grassland landscape. In this landscape, kites favored upland trees and shrubs, avoiding their more typical riparian forest association elsewhere in the species' range. Compared to random conditions, kites selected nest sites with high tree density and more closed canopy in the surrounding area. Mississippi Kite selection was not related to food abundance but could be explained by the presence of superpredators (i.e., hawks and owls) selecting riparian woodland for their nests. Nest survival declined with proximity to superpredator nesting sites. Overall, our study demonstrates how landscape structure and superior predators shapes predation risk for subordinate predators. Our results emphasize the importance of spatial heterogeneity in presenting opportunities for subordinate predators to coexist in a landscape with important superpredators.

  13. Monochromatic X-ray-induced thermal effect on four-reflection “nested” meV-monochromators: dynamical diffraction theory and finite-element analysis

    NASA Astrophysics Data System (ADS)

    Hu, Ling-Fei; Gao, Li-Dan; Li, Zhen-Jie; Wang, Shan-Feng; Sheng, Wei-Fan; Liu, Peng; Xu, Wei

    2015-09-01

    The high energy resolution monochromator (HRM) is widely used in inelastic scattering programs to detect phonons with energy resolution, down to the meV level. Although the large amount of heat from insertion devices can be reduced by a high heat-load monochromator, the unbalanced heat load on the inner pair of crystals in a nested HRM can affect its overall performance. Here, a theoretical analysis of the unbalanced heat load using dynamical diffraction theory and finite element analysis is presented. By utilizing the ray-tracing method, the performance of different HRM nesting configurations is simulated. It is suggested that the heat balance ratio, energy resolution, and overall spectral transmission efficiency are the figures of merit for evaluating the performance of nested HRMs. Although the present study is mainly focused on nested HRMs working at 57Fe nuclear resonant energy at 14.4 keV, it is feasible to extend this to other nested HRMs working at different energies.

  14. Are Nested Networks More Robust to Disturbance? A Test Using Epiphyte-Tree, Comensalistic Networks

    PubMed Central

    Piazzon, Martín; Larrinaga, Asier R.; Santamaría, Luis

    2011-01-01

    Recent research on ecological networks suggests that mutualistic networks are more nested than antagonistic ones and, as a result, they are more robust against chains of extinctions caused by disturbances. We evaluate whether mutualistic networks are more nested than comensalistic and antagonistic networks, and whether highly nested, host-epiphyte comensalistic networks fit the prediction of high robustness against disturbance. A review of 59 networks including mutualistic, antagonistic and comensalistic relationships showed that comensalistic networks are significantly more nested than antagonistic and mutualistic networks, which did not differ between themselves. Epiphyte-host networks from old-growth forests differed from those from disturbed forest in several topological parameters based on both qualitative and quantitative matrices. Network robustness increased with network size, but the slope of this relationship varied with nestedness and connectance. Our results indicate that interaction networks show complex responses to disturbances, which influence their topology and indirectly affect their robustness against species extinctions. PMID:21589931

  15. Turnover and dispersal of prairie falcons in southwestern Idaho

    USGS Publications Warehouse

    Lehman, Robert N.; Steenhof, Karen; Carpenter, L.B.; Kochert, Michael N.

    2000-01-01

    We studied Prairie Falcon (Falco mexicanus) breeding dispersal, natal dispersal, and turnover at nesting areas in the Snake River Birds of Prey National Conservation Area (NCA) from 1971- 95. Of 61 nesting areas where falcons identified one year were known to be present or absent the following year, 57% had a different falcon. This turnover rate was 2-3 times higher than that reported elsewhere for large falcons, and may have been related to high nesting densities in the NCA. Turnover at nesting areas was independent of nesting success in the previous year, but was significantly higher for females nesting on large cliffs. Mean distance between natal and breeding locations for 26 falcons banded as nestlings and later encountered as nesting adults was 8.9 km. Natal dispersal distances were similar for males and females, but more than twice as many males marked as nestlings were later encountered nesting in the NCA. Fourteen adult falcons found on different nesting areas in successive years moved an average of 1.5 km between nesting areas; males dispersed significantly farther than females. Natal and breeding dispersal distances in the NCA were lower than those reported for Prairie Falcons in other study areas. Only four falcons banded as nestlings were found outside NCA boundaries during the breeding period, and only one of these birds was known to be occupying a nesting area. We encountered no falcons banded outside the NCA occupying nesting areas in the NCA during this study.

  16. A nested-PCR with an Internal Amplification Control for the detection and differentiation of Bartonella henselae and B. clarridgeiae: An examination of cats in Trinidad

    PubMed Central

    Rampersad, Joanne N; Watkins, John D; Samlal, Michael S; Deonanan, Raymond; Ramsubeik, Shalini; Ammons, David R

    2005-01-01

    Background Bartonella species are bacterial blood parasites of animals capable of causing disease in both animals and man. Cat-Scratch Disease (CSD) in humans is caused mainly by Bartonella henselae and is acquired from the cat, which serves as a reservoir for the bacteria. A second species, B. clarridgeiae is also implicated in the disease. Diagnosis of Bartonellosis by culture requires a week or more of incubation on enriched media containing blood, and recovery is often complicated by faster growing contaminating bacteria and fungi. PCR has been explored as an alternative to culture for both the detection and species identification of Bartonella, however sensitivity problems have been reported and false negative reactions due to blood inhibitors have not generally been addressed in test design. Methods A novel, nested-PCR was designed for the detection of Bartonella henselae and B. clarridgeiae based on the strategy of targeting species-specific size differences in the 16S-23S rDNA intergenic regions. An Internal Amplification Control was used for detecting PCR inhibition. The nested-PCR was utilized in a study on 103 blood samples from pet and stray cats in Trinidad. Results None of the samples were positive by primary PCR, but the Nested-PCR detected Bartonella in 32/103 (31%) cats where 16 were infected with only B. henselae, 13 with only B. clarridgeiae and 3 with both species. Of 22 stray cats housed at an animal shelter, 13 (59%) were positive for either or both species, supporting the reported increased incidence of Bartonella among feral cats. Conclusion The usefulness of a single PCR for the detection of Bartonella henselae and B. clarridgeiae in the blood of cats is questionable. A nested-PCR offers increased sensitivity over a primary PCR and should be evaluated with currently used methods for the routine detection and speciation of Bartonella henselae and B. clarridgeiae. In Trinidad, B. henselae and B. clarridgeiae are the predominant species in cats and infection appears highest with stray cats, however B. clarridgeiae may be present at levels similar to that of B. henselae in the pet population. PMID:16098227

  17. House sparrows benefit from the conservation of white storks

    NASA Astrophysics Data System (ADS)

    Kosicki, Jakub Z.; Sparks, Tim H.; Tryjanowski, Piotr

    2007-05-01

    As with many farmland bird species, the house sparrow Passer domesticus is declining in Europe, mainly due to intensification of agriculture reducing nest sites and food supplies. During 2002-2005, we studied the population size and nest site characteristics of house sparrows breeding within white stork Ciconia ciconia nests in a large area of agricultural landscape within western Poland. To explain sparrow density within stork nests, we examined characteristics of white stork nests (position, age, productivity) and the farm type around the nest. House sparrow density was greatest in the longest established (and hence larger) white stork nests located on traditionally managed farms. Two recent changes appear to have adverse effects on house sparrows. The first is the intensification of farming and the second is active management of white stork nests on electric poles to reduce nest size and thus avoid both disruption to the electrical supply and electrocution of white storks. Because the white stork has such a high profile in Poland, there are numerous schemes to conserve and enhance this species. In conclusion, we clearly show that protecting one species can have valuable, although unplanned, benefits to another species of conservation interest, the house sparrow.

  18. House sparrows benefit from the conservation of white storks.

    PubMed

    Kosicki, Jakub Z; Sparks, Tim H; Tryjanowski, Piotr

    2007-05-01

    As with many farmland bird species, the house sparrow Passer domesticus is declining in Europe, mainly due to intensification of agriculture reducing nest sites and food supplies. During 2002-2005, we studied the population size and nest site characteristics of house sparrows breeding within white stork Ciconia ciconia nests in a large area of agricultural landscape within western Poland. To explain sparrow density within stork nests, we examined characteristics of white stork nests (position, age, productivity) and the farm type around the nest. House sparrow density was greatest in the longest established (and hence larger) white stork nests located on traditionally managed farms. Two recent changes appear to have adverse effects on house sparrows. The first is the intensification of farming and the second is active management of white stork nests on electric poles to reduce nest size and thus avoid both disruption to the electrical supply and electrocution of white storks. Because the white stork has such a high profile in Poland, there are numerous schemes to conserve and enhance this species. In conclusion, we clearly show that protecting one species can have valuable, although unplanned, benefits to another species of conservation interest, the house sparrow.

  19. Field evaluation of the photo-induced electron transfer fluorogenic primers (PET) real-time PCR for the detection of Plasmodium falciparum in Tanzania.

    PubMed

    Talundzic, Eldin; Maganga, Mussa; Masanja, Irene M; Peterson, David S; Udhayakumar, Venkatachalam; Lucchi, Naomi W

    2014-01-27

    Accurate diagnosis of malaria infections remains challenging, especially in the identification of submicroscopic infections. New molecular diagnostic tools that are inexpensive, sensitive enough to detect low-level infections and suitable in laboratory settings of resource-limited countries are required for malaria control and elimination programmes. Here the diagnostic potential of a recently developed photo-induced electron transfer fluorogenic primer (PET) real-time polymerase chain reaction (PCR) called PET-PCR was investigated. This study aimed to (i) evaluate the use of this assay as a method for the detection of both Plasmodium falciparum and other Plasmodium species infections in a developing country's diagnostic laboratory; and, (ii) determine the assay's sensitivity and specificity compared to a nested 18S rRNA PCR. Samples used in this study were obtained from a previous study conducted in the region of Iringa, Tanzania. A total of 303 samples from eight health facilities in Tanzania were utilized for this evaluation. All samples were screened using the multiplex PET-PCR assay designed to detect Plasmodium genus and P. falciparum initially in laboratory in Tanzania and then repeated at a reference laboratory at the CDC in the USA. Microscopy data was available for all the 303 samples. A subset of the samples were tested in a blinded fashion to find the sensitivity and specificity of the PET-PCR compared to the nested 18S rRNA PCR. Compared to microscopy, the PET-PCR assay was 59% more sensitive in detecting P. falciparum infections. The observed sensitivity and specificity were 100% (95% confidence interval (CI0.95) = 94-100%) and (CI0.95 = 96-100%), respectively, for the PET-PCR assay when compared to nested 18S rRNA PCR. When compared to 18S rRNA PCR, microscopy had a low sensitivity of 40% (CI0.95 = 23-61%) and specificity of 100% (CI0.95 = 96-100%). The PET-PCR results performed in the field laboratory in Tanzania were in 100% concordance with the results obtained at the reference laboratory in the USA. The PET-PCR is a new molecular diagnostic tool with similar performance characteristics as commonly used PCR methods that is less expensive, easy to use, and amiable to large scale-surveillance studies in developing country settings.

  20. Flexible social organization and high incidence of drifting in the sweat bee, Halictus scabiosae.

    PubMed

    Ulrich, Yuko; Perrin, Nicolas; Chapuisat, Michel

    2009-04-01

    The very diverse social systems of sweat bees make them interesting models to study social evolution. Here we focus on the dispersal behaviour and social organization of Halictus scabiosae, a common yet poorly known species of Europe. By combining field observations and genetic data, we show that females have multiple reproductive strategies, which generates a large diversity in the social structure of nests. A detailed microsatellite analysis of 60 nests revealed that 55% of the nests contained the offspring of a single female, whereas the rest had more complex social structures, with three clear cases of multiple females reproducing in the same nest and frequent occurrence of unrelated individuals. Drifting among nests was surprisingly common, as 16% of the 122 nests in the overall sample and 44% of the nests with complex social structure contained females that had genotypes consistent with being full-sisters of females sampled in other nests of the population. Drifters originated from nests with an above-average productivity and were unrelated to their nestmates, suggesting that drifting might be a strategy to avoid competition among related females. The sex-specific comparison of genetic differentiation indicated that dispersal was male-biased, which would reinforce local resource competition among females. The pattern of genetic differentiation among populations was consistent with a dynamic process of patch colonization and extinction, as expected from the unstable, anthropogenic habitat of this species. Overall, our data show that H. scabiosae varies greatly in dispersal behaviour and social organization. The surprisingly high frequency of drifters echoes recent findings in wasps and bees, calling for further investigation of the adaptive basis of drifting in the social insects.

  1. Temporal changes in artificial light exposure of marine turtle nesting areas.

    PubMed

    Kamrowski, Ruth L; Limpus, Col; Jones, Rhondda; Anderson, Sharolyn; Hamann, Mark

    2014-08-01

    Artificial light at night poses a significant threat to multiple taxa across the globe. In coastal regions, artificial lighting close to marine turtle nesting beaches is disruptive to their breeding success. Prioritizing effective management of light pollution requires an understanding of how the light exposure of nesting areas changes over time in response to changing temporal and spatial distributions of coastal development. We analyzed multitemporal, satellite night-light data, in combination with linear mixed model analysis, to determine broadscale changes in artificial light exposure at Australian marine turtle nesting areas between 1993 and 2010. We found seven marine turtle management units (MU), from five species, have experienced significant increases in light exposure over time, with flatback turtles nesting in east Australia experiencing the fastest increases. The remaining 12 MUs showed no significant change in light exposure. Unchanging MUs included those previously identified as having high exposure to light pollution (located in western Australia and southern Queensland), indicating that turtles in these areas have been potentially exposed to high light levels since at least the early nineties. At a finer geographic scale (within-MU), nine MUs contained nesting areas with significant increases in light exposure. These nesting areas predominantly occurred close to heavily industrialized coastal areas, thus emphasizing the importance of rigorous light management in industry. Within all MUs, nesting areas existed where light levels were extremely low and/or had not significantly increased since 1993. With continued coastal development, nesting females may shift to these darker/unchanging 'buffer' areas in the future. This is valuable information that informs our understanding of the capacity and resilience of marine turtles faced with coastal development: an understanding that is essential for effective marine turtle conservation. © 2013 John Wiley & Sons Ltd.

  2. Occupancy modeling reveals territory-level effects of nest boxes on the presence, colonization, and persistence of a declining raptor in a fruit-growing region.

    PubMed

    Shave, Megan E; Lindell, Catherine A

    2017-01-01

    Nest boxes for predators in agricultural regions are an easily implemented tool to improve local habitat quality with potential benefits for both conservation and agriculture. The potential for nest boxes to increase raptor populations in agricultural regions is of particular interest given their positions as top predators. This study examined the effects of cherry orchard nest boxes on the local breeding population of a declining species, the American Kestrel (Falco sparverius), in a fruit-growing region of Michigan. During the 2013-2016 study, we added a total of 23 new nest boxes in addition to 24 intact boxes installed previously; kestrels used up to 100% of our new boxes each season. We conducted temporally-replicated surveys along four roadside transects divided into 1.6 km × 500 m sites. We developed a multi-season occupancy model under a Bayesian framework and found that nest boxes had strong positive effects on first-year site occupancy, site colonization, and site persistence probabilities. The estimated number of occupied sites increased between 2013 and 2016, which correlated with the increase in number of sites with boxes. Kestrel detections decreased with survey date but were not affected by time of day or activity at the boxes themselves. These results indicate that nest boxes determined the presence of kestrels at our study sites and support the conclusion that the local kestrel population is likely limited by nest site availability. Furthermore, our results are highly relevant to the farmers on whose properties the boxes were installed, for we can conclude that installing a nest box in an orchard resulted in a high probability of kestrels occupying that orchard or the areas adjacent to it.

  3. Nest spacing, habitat selection, and behavior of waterfowl on Miller Lake Island, North Dakota

    USGS Publications Warehouse

    Lokemoen, J.T.; Duebbert, H.F.; Sharp, D.E.

    1984-01-01

    The nesting behavior of a concentration of nesting mallards (Anas platyrhynchos) and gadwalls (A. strepera) was studied on a 4.5-ha island in Miller Lake, North Dakota, in 1977. A single 0.59-ha clump of thick shrub contained 225 simultaneously active mallard nests on 10 May. During the peak nesting period, mallard nests were spaced an average of 2.7 m from conspecifics. Active nests of all species peaked at 327, spaced an average of 2.1 m apart. Nests were clustered in thick shrub with moderate numbers in open shrub and few in grassland. Nest placement was significantly related to the amount of vegetative screening although mammalian predators were absent on the island. Cover density appeared to be important to the nesting hens as vegetation screened nests from potential avian predators and from harassment by other conspecific nesting hens and drakes. Mallards and gadwalls nesting on this island used wetlands in >100 km2 around Miller Lake. During daylight hours in late April 1978, an average of 4.9 mallards/minute arrived at the island; a peak of 17.2 mallards/minute arrived at 0800. In late April 1978, as many as 26 mallard pairs/ha occurred on favored wetlands and behavioral aggression was intense. Yet, most mallard hens maintained adequate space to acquire food and other requisites. Mallards and gadwalls were sufficiently adaptable to successfully exploit the Miller Lake island and environments that resulted in high reproductive success. Other local breeding Anatidae because of behavioral constraints were unable to exploit the same situation.

  4. Convective and microphysics parameterization impact on simulating heavy rainfall in Semarang (case study on February 12th, 2015)

    NASA Astrophysics Data System (ADS)

    Faridatussafura, Nurzaka; Wandala, Agie

    2018-05-01

    The meteorological model WRF-ARW version 3.8.1 is used for simulating the heavy rainfall in Semarang that occurred on February 12th, 2015. Two different convective schemes and two different microphysics scheme in a nested configuration were chosen. The sensitivity of those schemes in capturing the extreme weather event has been tested. GFS data were used for the initial and boundary condition. Verification on the twenty-four hours accumulated rainfall using GSMaPsatellite data shows that Kain-Fritsch convective scheme and Lin microphysics scheme is the best combination scheme among the others. The combination also gives the highest success ratio value in placing high intensity rainfall area. Based on the ROC diagram, KF-Lin shows the best performance in detecting high intensity rainfall. However, the combination still has high bias value.

  5. Nest predation of Abert's Towhees by coachwhips and roadrunners

    Treesearch

    Deborah M. Finch

    1981-01-01

    Predation by snakes has been implicated as a major cause of high nesting mortality in many passerine birds (e.g., Willis 1972, Best 1978, Nolan 1978), but predation is rarely observed (e.g., Snow 1962, Lill 1974). Snakes and other predators typically consume the entire contents of a nest during one visit (e.g., Nolan 1978). Nestling starvation caused by sibling...

  6. Long-eared owls nesting in Badlands National Park

    Treesearch

    Deborah D. Paulson; Carolyn Hull Sieg

    1985-01-01

    Long-eared Owls nest at high densities locally over the Great Plains where suitable habitat is limited (Bent 1938), yet , according to Whitney et al. (1978), this species is rare to uncommon in South Dakota. Especially west of the Missouri River, few nesting records have been reported. This paper reports the occurrence of Long-eared Owls in the Badlands National Park...

  7. Landscape patterns around Northern Spotted Owl (Strix occidentalis caurina) nest sites in Oregon's central Coast Ranges

    Treesearch

    J. P. Perkins; J. A. Thrailkill; W. J. Ripple; K. T. Hershey

    1997-01-01

    We investigated landscape characteristics around 41 Northern Spotted Owl (Strix occidentalis caurina) nest sites to assess habitat proportions and patterns on this highly fragmented landscape in the central Coast Ranges of Oregon. We compared the proportion of seven forest cover-types between nest sites and random sites at plot sizes of 112 ha, 456...

  8. Biotic and abiotic factors affect the nest environment of embryonic leatherback turtles, Dermochelys coriacea.

    PubMed

    Wallace, Bryan P; Sotherland, Paul R; Spotila, James R; Reina, Richard D; Franks, Bryan F; Paladino, Frank V

    2004-01-01

    Clutches of leatherback turtles, Dermochelys coriacea, have lower hatching success than those of other sea turtles, but causes of high embryonic mortality are unknown. We measured characteristics of clutches along with spatial and temporal changes in PO(2) and temperature during incubation to determine the extent to which they affected the developmental environment of leatherback embryos. Minimum PO(2) in nests decreased as both the total number and mass of metabolizing embryos increased. Increases in both the number and mass of metabolizing embryos caused an increase in maximum nest temperature. However, neither PO(2) nor temperature was correlated with hatching success. Our measurements of relatively high nest PO(2) (lowest 17.1 kPa or 16.9% O(2)) indicate that hypoxia apparently does not cause the low hatching success of leatherback clutches. Oxygen partial pressure increased and temperature decreased from the center toward the periphery of leatherback nests. We inferred from these measurements that positions of eggs within nests vary in quality and potentially affect overall developmental success of entire clutches. The large metabolic mass of leatherback clutches and limits to gas flux imposed by the sand create a situation in which leatherback embryos collectively affect their own environment.

  9. Identification and properties of steroid-binding proteins in nesting Chelonia mydas plasma.

    PubMed

    Ikonomopoulou, M P; Bradley, A J; Whittier, J M; Ibrahim, K

    2006-11-01

    We report for the first time the presence of a sex steroid-binding protein in the plasma of green sea turtles Chelonia mydas, which provides an insight into reproductive status. A high affinity, low capacity sex hormone steroid-binding protein was identified in nesting C. mydas and its thermal profile was established. In nesting C. mydas testosterone and oestradiol bind at 4 degrees C with high affinity (K (a) = 1.49 +/- 0.09 x 10(9) M(-1); 0.17 +/- 0.02 x 10(7) M(-1)) and low binding capacity (B (max) = 3.24 +/- 0.84 x 10(-5) M; 0.33 +/- 0.06 x 10(-4) M). The binding affinity and capacity of testosterone at 23 and 36 degrees C, respectively were similar to those determined at 4 degrees C. However, oestradiol showed no binding activity at 36 degrees C. With competition studies we showed that oestradiol and oestrone do not compete for binding sites. Furthermore, in nesting C. mydas plasma no high-affinity binding was observed for adrenocortical steroids (cortisol and corticosterone) and progesterone. Our results indicate that in nesting C. mydas plasma temperature has a minimal effect on the high-affinity binding of testosterone to sex steroid-binding protein, however, the high affinity binding of oestradiol to sex steroid-binding protein is abolished at a hypothetically high (36 degrees C) sea/ambient/body temperature. This suggests that at high core body temperatures most of the oestradiol becomes biologically available to the tissues rather than remaining bound to a high-affinity carrier.

  10. Evaluating nest supplementation as a recovery strategy for the endangered rodents of the Florida Keys

    USGS Publications Warehouse

    Cove, Michael V.; Simons, Theodore R.; Gardner, Beth; Maurer, Andrew S.; O'Connell, Allan F.

    2017-01-01

    The Key Largo woodrat (Neotoma floridana smalli) and Key Largo cotton mouse (Peromyscus gossypinus allapaticola) are federally endangered subspecies endemic to the tropical hardwood hammocks of Key Largo, Florida. Woodrats are considered generalists in habitat and diet, yet a steady decline in natural stick nests and capture rates over the past several decades suggests that they are limited by the availability of nesting habitat due to habitat loss and fragmentation. The more specialized Key Largo cotton mouse appears to rely on old growth hammock, a habitat type that is rare following past land clearing. In 2004, the U.S. Fish and Wildlife Service started building supplemental nest structures to restore habitat quality and connectivity for these endangered rodents, but nest use requires evaluation. We used camera traps and occupancy models to evaluate the factors influencing woodrat and cotton mouse use of the supplemental nests. We detected woodrats at 65 and cotton mice at 175 of 284 sampled nest structures, with co-occurrence at 38 nests. Woodrat nest use followed a gradient from low nest use in the north to high nest use in the south, which might relate to the proximity of free-ranging domestic cat (Felis catus) colonies in residential developments. Cotton mouse nest use, however, was related positively to mature hammock and related negatively to disturbed areas (e.g. scarified lands). The two species occurred independently of each other. Stick-stacking behavior was observed at supplemental nests and, although it was correlated with detection of woodrats, it was not a strong predictor of their occurrence. We suggest that nest supplementation can be an important tool for species recovery as habitat quality continues to improve with succession.

  11. Interactive effects between nest microclimate and nest vegetation structure confirm microclimate thresholds for Lesser Prairie-Chicken nest survival

    USGS Publications Warehouse

    Grisham, Blake A.; Godar, Alixandra J.; Boal, Clint W.; Haukos, David A.

    2016-01-01

    The range of Lesser Prairie-Chickens (Tympanuchus pallidicinctus) spans 4 unique ecoregions along 2 distinct environmental gradients. The Sand Shinnery Oak Prairie ecoregion of the Southern High Plains of New Mexico and Texas is environmentally isolated, warmer, and more arid than the Short-Grass, Sand Sagebrush, and Mixed-Grass Prairie ecoregions in Colorado, Kansas, Oklahoma, and the northeast panhandle of Texas. Weather is known to influence Lesser Prairie-Chicken nest survival in the Sand Shinnery Oak Prairie ecoregion; regional variation may also influence nest microclimate and, ultimately, survival during incubation. To address this question, we placed data loggers adjacent to nests during incubation to quantify temperature and humidity distribution functions in 3 ecoregions. We developed a suite of a priori nest survival models that incorporated derived microclimate parameters and visual obstruction as covariates in Program MARK. We monitored 49 nests in Mixed-Grass, 22 nests in Sand Shinnery Oak, and 30 nests in Short-Grass ecoregions from 2010 to 2014. Our findings indicated that (1) the Sand Shinnery Oak Prairie ecoregion was hotter and drier during incubation than the Mixed- and Short-Grass ecoregions; (2) nest microclimate varied among years within ecoregions; (3) visual obstruction was positively associated with nest survival; but (4) daily nest survival probability decreased by 10% every half-hour when temperature was greater than 34°C and vapor pressure deficit was less than −23 mmHg during the day (about 0600–2100 hours). Our major finding confirmed microclimate thresholds for nest survival under natural conditions across the species' distribution, although Lesser Prairie-Chickens are more likely to experience microclimate conditions that result in nest failures in the Sand Shinnery Oak Prairie ecoregion. The species would benefit from identification of thermal landscapes and management actions that promote cooler, more humid nest microclimates.

  12. Methods for reducing singly reflected rays on the Wolter-I focusing mirrors of the FOXSI rocket experiment

    NASA Astrophysics Data System (ADS)

    Buitrago-Casas, Juan Camilo; Elsner, Ronald; Glesener, Lindsay; Christe, Steven; Ramsey, Brian; Courtade, Sasha; Ishikawa, Shin-nosuke; Narukage, Noriyuki; Turin, Paul; Vievering, Juliana; Athiray, P. S.; Musset, Sophie; Krucker, Säm.

    2017-08-01

    In high energy solar astrophysics, imaging hard X-rays by direct focusing offers higher dynamic range and greater sensitivity compared to past techniques that used indirect imaging. The Focusing Optics X-ray Solar Imager (FOXSI) is a sounding rocket payload that uses seven sets of nested Wolter-I figured mirrors together with seven high-sensitivity semiconductor detectors to observe the Sun in hard X-rays through direct focusing. The FOXSI rocket has successfully flown twice and is funded to fly a third time in summer 2018. The Wolter-I geometry consists of two consecutive mirrors, one paraboloid and one hyperboloid, that reflect photons at grazing angles. Correctly focused X-rays reflect once per mirror segment. For extended sources, like the Sun, off-axis photons at certain incident angles can reflect on only one mirror and still reach the focal plane, generating a background pattern of singly reflected rays (i.e., ghost rays) that can limit the sensitivity of the observation to faint, focused sources. Understanding and mitigating the impact of the singly reflected rays on the FOXSI optical modules will maximize the instruments' sensitivity to background-limited sources. We present an analysis of the FOXSI singly reflected rays based on ray-tracing simulations and laboratory measurements, as well as the effectiveness of different physical strategies to reduce them.

  13. The Focusing Optics X-ray Solar Imager

    NASA Astrophysics Data System (ADS)

    Glesener, Lindsay; Krucker, S.; Christe, S.; Turin, P.; McBride, S.

    2009-01-01

    The Focusing Optics X-ray Solar Imager (FOXSI) is a NASA Low Cost Access to Space sounding rocket payload scheduled to fly in late 2010 to observe hard X-ray emission (HXR) from the quiet Sun. Particle acceleration in small "nanoflares" in the quiet Sun is thought to play an important role in the heating of the corona to millions of degrees Kelvin. FOXSI HXR observations of these flares will provide first estimates of the non-thermal energy content in small flares from the quiet Sun. Imaging nanoflares requires high energy sensitivity and a large dynamic range. To date, the most sensitive HXR images are made using a rotating modulating collimator aboard the Reuven Ramaty High Energy Spectroscopic Imager satellite (RHESSI). However, the rotating modulation technique is intrinsically limited in sensitivity and dynamic range. The focusing optics of FOXSI will achieve a sensitivity 100 times better than that of RHESSI at energies around 10 keV. FOXSI uses nested-shell, grazing-angle optics and silicon strip detectors to achieve an angular resolution of 12" (FWHM) and 1 keV energy resolution. FOXSI will observe the quiet Sun in the 4 to 15 keV range for 5 minutes. The focusing optics technique developed by FOXSI will prove useful to future solar HXR observing missions, especially those interested in imaging faint HXR emission from particle acceleration regions in the corona.

  14. Ectoparasitism and the role of green nesting material in the European starling

    USGS Publications Warehouse

    Fauth, P.T.; Krementz, D.G.; Hines, J.E.

    1991-01-01

    The use of green nesting material is wide-spread among birds. Recent evidence suggests that birds use secondary chemicals contained in green plants to control ectoparasites. We manipulated green nesting material and ectoparasites of European starlings (Sturnus vulgaris ) to test two hypotheses: (1) ectoparasites adversely affect prefledging survival and morphometrics or postfledging survival, and (2) green nesting material ameliorates the effects of ectoparasites. We recorded fat score, number of scabs, tarsal length, body mass, and hematocrit level on each nestling 17 days after hatching. We also fitted each nestling with unique patagial tags and resighted the starlings for 6-8 weeks after fledging to estimate survival and sighting rates. Nests devoid of green nesting material and dusted with the insecticide, carbaryl, had fewer high ectoparasite infestations, and nestlings had significantly lower scab scores, and significantly higher body masses than nestlings in undusted boxes. However, there was no difference in postfledging survival between birds from carbaryl-treated and undusted nests.

  15. Impacts of Water Levels on Breeding Canada Geese and Methods for Mitigation and Management in the Southern Flathead Valley, Montana, 1983-1987 Final Report.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mackey, Dennis L.; Gregory, Shari K.; Matthews, William C. Jr.

    Kerr Hydroelectric Dam is located at the south end of Flathead Lake, controls water levels on the lake and the Flathead River below the dam, and is currently operated as a load control facility. Current operation of Kerr Dam creates the greatest yearly water level fluctuations on both the lake and river during the Canada goose (Branta canadensis moffitti) brood and nesting period. Data collected from 1980-1982 indicated that goose nest numbers on the river were lower than during the 1950's, and that brood habitat on the lake may be limiting the goose population there. Our study was conducted frommore » 1983-1987 to determine the effects of Kerr Dam operation on Canada goose populations and habitat on the south half of Flathead Lake and the Flathead River, and to formulate management and mitigation recommendations. Nesting geese on the river appeared to be negatively affected by a lack of nest sites free from predators, and responded to available artificial nest structures with an increase in nest numbers and nesting success. Under current dam operation, river channel depths and widths do not discourage access to nesting islands by mammalian predators during some years and high predation on ground nests occurs. Intensively used brood areas on the lake and river were identified and described. Brood habitat on the lake was lower in quality and quantity than on the river due to dam operations. Gosling mortality on the lake was high, almost 2 times higher than on the river. Lake broods expended more energy obtaining food than river broods. Losses of brood habitat in the form of wet meadow marshes were documented and mitigation options developed. Management/mitigation alternatives and monitoring methods for nesting and brooding geese were identified.« less

  16. The Urban Nest.

    ERIC Educational Resources Information Center

    Pollowy, Anne-Marie

    In this book, the many factors necessary to make city spaces more child-sensitive are considered. Particular attention is given to dwellings and play spaces. Child development as it relates to the spatial environment and the child's use of various spaces (houses, apartments, service areas, playgrounds) at different ages are investigated. The…

  17. Reproductive ecology of the Maui Parrotbill

    USGS Publications Warehouse

    Simon, John C.; Pratt, T.K.; Berlin, Kim E.; Kowalsky, James R.

    2000-01-01

    The endangered Maui Parrotbill (Pseudonestor xanthophrys) is an excavating, insectivorous Hawaiian honeycreeper endemic to the high elevation rain forests of east Maui, Hawaii. From March 1994 to June 1997, we studied various aspects of their breeding ecology. We color-banded 18 individuals, located and monitored 9 active nests, and took behavioral data during 440 hrs of nest observation. Both members of a pair maintained a year-round, all-purpose territory that included nest sites and food resources. Maui Parrotbill were monogamous within and between years; we found no evidence of polyandry, polygyny, or helpers at the nest. Nests were cup-shaped, composed mainly of lichen interlaced with small twigs, and positioned in the outer canopy forks of mature ohia (Metrosideros polymorpha) trees. Modal clutch size was one. Females performed most nest construction and all incubation and brooding; males provisioned females and assisted in feeding nestlings after their fourth day. Fledglings depended on parental care for 5-8 months, during which their bill strength increased and foraging skills improved. We calculated the overall nest success rate by the Mayfield Method as 0.42 for the 1995/1996 and 1996/1997 breeding seasons combined. Nest failure and fledgling disappearance coincided with events of high rainfall. Their breeding ecology most closely resembled the Akiapolaau (Hemignathus munroi), another excavating, insectivorous Hawaiian honeycreeper found on Hawaii Island. As with the Akiapolaau, the threat of extinction is persistent and results from both the constraints of inherent life history traits and artificial ecological changes. We advocate the protection and expansion of habitable forest areas and an ongoing program to monitor and mitigate the effects of invasive species.

  18. Evaluation of Trail-Cameras for Analyzing the Diet of Nesting Raptors Using the Northern Goshawk as a Model

    PubMed Central

    García-Salgado, Gonzalo; Rebollo, Salvador; Pérez-Camacho, Lorenzo; Martínez-Hesterkamp, Sara; Navarro, Alberto; Fernández-Pereira, José-Manuel

    2015-01-01

    Diet studies present numerous methodological challenges. We evaluated the usefulness of commercially available trail-cameras for analyzing the diet of Northern Goshawks (Accipiter gentilis) as a model for nesting raptors during the period 2007–2011. We compared diet estimates obtained by direct camera monitoring of 80 nests with four indirect analyses of prey remains collected from the nests and surroundings (pellets, bones, feather-and-hair remains, and feather-hair-and-bone remains combined). In addition, we evaluated the performance of the trail-cameras and whether camera monitoring affected Goshawk behavior. The sensitivity of each diet-analysis method depended on prey size and taxonomic group, with no method providing unbiased estimates for all prey sizes and types. The cameras registered the greatest number of prey items and were probably the least biased method for estimating diet composition. Nevertheless this direct method yielded the largest proportion of prey unidentified to species level, and it underestimated small prey. Our trail-camera system was able to operate without maintenance for longer periods than what has been reported in previous studies with other types of cameras. Initially Goshawks showed distrust toward the cameras but they usually became habituated to its presence within 1–2 days. The habituation period was shorter for breeding pairs that had previous experience with cameras. Using trail-cameras to monitor prey provisioning to nests is an effective tool for studying the diet of nesting raptors. However, the technique is limited by technical failures and difficulties in identifying certain prey types. Our study also shows that cameras can alter adult Goshawk behavior, an aspect that must be controlled to minimize potential negative impacts. PMID:25992956

  19. Evaluation of trail-cameras for analyzing the diet of nesting raptors using the Northern Goshawk as a model.

    PubMed

    García-Salgado, Gonzalo; Rebollo, Salvador; Pérez-Camacho, Lorenzo; Martínez-Hesterkamp, Sara; Navarro, Alberto; Fernández-Pereira, José-Manuel

    2015-01-01

    Diet studies present numerous methodological challenges. We evaluated the usefulness of commercially available trail-cameras for analyzing the diet of Northern Goshawks (Accipiter gentilis) as a model for nesting raptors during the period 2007-2011. We compared diet estimates obtained by direct camera monitoring of 80 nests with four indirect analyses of prey remains collected from the nests and surroundings (pellets, bones, feather-and-hair remains, and feather-hair-and-bone remains combined). In addition, we evaluated the performance of the trail-cameras and whether camera monitoring affected Goshawk behavior. The sensitivity of each diet-analysis method depended on prey size and taxonomic group, with no method providing unbiased estimates for all prey sizes and types. The cameras registered the greatest number of prey items and were probably the least biased method for estimating diet composition. Nevertheless this direct method yielded the largest proportion of prey unidentified to species level, and it underestimated small prey. Our trail-camera system was able to operate without maintenance for longer periods than what has been reported in previous studies with other types of cameras. Initially Goshawks showed distrust toward the cameras but they usually became habituated to its presence within 1-2 days. The habituation period was shorter for breeding pairs that had previous experience with cameras. Using trail-cameras to monitor prey provisioning to nests is an effective tool for studying the diet of nesting raptors. However, the technique is limited by technical failures and difficulties in identifying certain prey types. Our study also shows that cameras can alter adult Goshawk behavior, an aspect that must be controlled to minimize potential negative impacts.

  20. Detection of Toxoplasma gondii oocysts in soils in northwestern China using a new semi-nested PCR assay.

    PubMed

    Wang, Meng; Meng, Peng; Ye, Qiang; Pu, Yuan-Hua; Yang, Xiao-Yu; Luo, Jian-Xun; Zhang, Nian-Zhang; Zhang, De-Lin

    2014-09-28

    Toxoplasma gondii is a zoonotic pathogen that can infect a range of animals and humans. Ingestion of T. gondii oocysts in soil is a significant transmission route for humans and animals acquiring toxoplasmosis. In the present study, we developed a new semi-nested PCR method to determine T. gondii oocysts distribution in soils in northwestern China. The one tube semi-nested PCR assay was developed to detect the oocysts of T. gondii in soil, targeting the repetitive 529 bp fragment of T. gondii genomic DNA. Then a total of 268 soil samples, including 148 samples from Gansu Province and 120 samples from Qinghai Province, northwestern China, were examined by the semi-nested PCR method. One third of the positive samples were sequenced. The sensitivity of the semi-nested PCR assay was 10(2)  T. gondii oocysts in 5 g soil sample. Investigation of soil samples from northwestern China showed that 34 out of 268 soil samples (12.69%) were T. gondii positive. Sequences of the partial 529 bp fragments varied from 0-1.2% among the sequenced samples. The prevalence of T. gondii oocysts in soil from cities (24/163) was slightly higher than that in soils from pasturing areas (10/105) (P = 0.21). Among the different regions in cities, the prevalence of T. gondii oocysts in soils from parks was 14.15%, whereas that in soils from schools was 19.05%. The present study firstly reported the prevalence of T. gondii oocysts in soils in northwest China using a novel semi-nested PCR assay, which provided baseline data for the effective prevention and control of toxoplasmosis in this region.

  1. Ocean regional circulation model sensitizes to resolution of the lateral boundary conditions

    NASA Astrophysics Data System (ADS)

    Pham, Van Sy; Hwang, Jin Hwan

    2017-04-01

    Dynamical downscaling with nested regional oceanographic models is an effective approach for forecasting operationally coastal weather and projecting long term climate on the ocean. Nesting procedures deliver the unwanted in dynamic downscaling due to the differences of numerical grid sizes and updating steps. Therefore, such unavoidable errors restrict the application of the Ocean Regional Circulation Model (ORCMs) in both short-term forecasts and long-term projections. The current work identifies the effects of errors induced by computational limitations during nesting procedures on the downscaled results of the ORCMs. The errors are quantitatively evaluated for each error source and its characteristics by the Big-Brother Experiments (BBE). The BBE separates identified errors from each other and quantitatively assess the amount of uncertainties employing the same model to simulate for both nesting and nested model. Here, we focus on discussing errors resulting from two main matters associated with nesting procedures. They should be the spatial grids' differences and the temporal updating steps. After the diverse cases from separately running of the BBE, a Taylor diagram was adopted to analyze the results and suggest an optimization intern of grid size and updating period and domain sizes. Key words: lateral boundary condition, error, ocean regional circulation model, Big-Brother Experiment. Acknowledgement: This research was supported by grants from the Korean Ministry of Oceans and Fisheries entitled "Development of integrated estuarine management system" and a National Research Foundation of Korea (NRF) Grant (No. 2015R1A5A 7037372) funded by MSIP of Korea. The authors thank the Integrated Research Institute of Construction and Environmental Engineering of Seoul National University for administrative support.

  2. Foraging habitat quality constrains effectiveness of artificial nest-site provisioning in reversing population declines in a colonial cavity nester.

    PubMed

    Catry, Inês; Franco, Aldina M A; Rocha, Pedro; Alcazar, Rita; Reis, Susana; Cordeiro, Ana; Ventim, Rita; Teodósio, Joaquim; Moreira, Francisco

    2013-01-01

    Among birds, breeding numbers are mainly limited by two resources of major importance: food supply and nest-site availability. Here, we investigated how differences in land-use and nest-site availability affected the foraging behaviour, breeding success and population trends of the colonial cavity-dependent lesser kestrel Falco naumanni inhabiting two protected areas. Both areas were provided with artificial nests to increase nest-site availability. The first area is a pseudo-steppe characterized by traditional extensive cereal cultivation, whereas the second area is a previous agricultural zone now abandoned or replaced by forested areas. In both areas, lesser kestrels selected extensive agricultural habitats, such as fallows and cereal fields, and avoided scrubland and forests. In the second area, tracked birds from one colony travelled significantly farther distances (6.2 km ± 1.7 vs. 1.8 km ± 0.4 and 1.9 km ± 0.6) and had significant larger foraging-ranges (144 km(2) vs. 18.8 and 14.8 km(2)) when compared to the birds of two colonies in the extensive agricultural area. Longer foraging trips were reflected in lower chick feeding rates, lower fledging success and reduced chick fitness. Availability and occupation of artificial nests was high in both areas but population followed opposite trends, with a positive increment recorded exclusively in the first area with a large proportion of agricultural areas. Progressive habitat loss around the studied colony in the second area (suitable habitat decreased from 32% in 1990 to only 7% in 2002) is likely the main driver of the recorded population decline and suggests that the effectiveness of bird species conservation based on nest-site provisioning is highly constrained by habitat quality in the surrounding areas. Therefore, the conservation of cavity-dependent species may be enhanced firstly by finding the best areas of remaining habitat and secondly by increasing the carrying capacity of high-quality habitat areas through safe nest-site provisioning.

  3. Foraging Habitat Quality Constrains Effectiveness of Artificial Nest-Site Provisioning in Reversing Population Declines in a Colonial Cavity Nester

    PubMed Central

    Catry, Inês; Franco, Aldina M. A.; Rocha, Pedro; Alcazar, Rita; Reis, Susana; Cordeiro, Ana; Ventim, Rita; Teodósio, Joaquim; Moreira, Francisco

    2013-01-01

    Among birds, breeding numbers are mainly limited by two resources of major importance: food supply and nest-site availability. Here, we investigated how differences in land-use and nest-site availability affected the foraging behaviour, breeding success and population trends of the colonial cavity-dependent lesser kestrel Falco naumanni inhabiting two protected areas. Both areas were provided with artificial nests to increase nest-site availability. The first area is a pseudo-steppe characterized by traditional extensive cereal cultivation, whereas the second area is a previous agricultural zone now abandoned or replaced by forested areas. In both areas, lesser kestrels selected extensive agricultural habitats, such as fallows and cereal fields, and avoided scrubland and forests. In the second area, tracked birds from one colony travelled significantly farther distances (6.2 km ±1.7 vs. 1.8 km ±0.4 and 1.9 km ±0.6) and had significant larger foraging-ranges (144 km2 vs. 18.8 and 14.8 km2) when compared to the birds of two colonies in the extensive agricultural area. Longer foraging trips were reflected in lower chick feeding rates, lower fledging success and reduced chick fitness. Availability and occupation of artificial nests was high in both areas but population followed opposite trends, with a positive increment recorded exclusively in the first area with a large proportion of agricultural areas. Progressive habitat loss around the studied colony in the second area (suitable habitat decreased from 32% in 1990 to only 7% in 2002) is likely the main driver of the recorded population decline and suggests that the effectiveness of bird species conservation based on nest-site provisioning is highly constrained by habitat quality in the surrounding areas. Therefore, the conservation of cavity-dependent species may be enhanced firstly by finding the best areas of remaining habitat and secondly by increasing the carrying capacity of high-quality habitat areas through safe nest-site provisioning. PMID:23484016

  4. Feeding rates, double brooding, nest reuse, and seasonal fecundity of eastern wood-pewees in the Missouri Ozarks

    Treesearch

    Sarah W. Kendrick; Frank R. III Thompson; Jennifer L. Reidy

    2014-01-01

    Despite being widespread and abundant, little is known about the breeding ecology and natural history of the Eastern Wood-Pewee (Contopus virens), in part because nests are often high in the canopy, difficult to view, and adults are monomorphic. We monitored nests of Eastern Wood-Pewees and recorded the feeding rate of nestlings by adults as part of...

  5. Comparative study of Gram stain, potassium hydroxide smear, culture and nested PCR in the diagnosis of fungal keratitis.

    PubMed

    Badiee, Parisa; Nejabat, Mahmood; Alborzi, Abdolvahab; Keshavarz, Fatemeh; Shakiba, Elaheh

    2010-01-01

    This study seeks to evaluate the efficacy and practicality of the molecular method, compared to the standard microbiological techniques for diagnosing fungal keratitis (FK). Patients with eye findings suspected of FK were enrolled for cornea sampling. Scrapings from the affected areas of the infected corneas were obtained and were divided into two parts: one for smears and cultures, and the other for nested PCR analysis. Of the 38 eyes, 28 were judged to have fungal infections based on clinical and positive findings in the culture, smear and responses to antifungal treatment. Potassium hydroxide, Gram staining, culture and nested PCR results (either positive or negative) matched in 76.3, 42.1, 68.4 and 81.6%, respectively. PCR is a sensitive method but due to the lack of sophisticated facilities in routine laboratory procedures, it can serve only complementarily and cannot replace conventional methods. Copyright © 2010 S. Karger AG, Basel.

  6. Should I stay or should I go? Hormonal control of nest abandonment in a long-lived bird, the Adélie penguin.

    PubMed

    Spée, Marion; Beaulieu, Michaël; Dervaux, Antoine; Chastel, Olivier; Le Maho, Yvon; Raclot, Thierry

    2010-11-01

    According to life-history theory, long-lived birds should favor their survival over the current reproductive attempt, when breeding becomes too costly. In seabirds, incubation is often associated with spontaneous long-term fasting. Below a threshold in body reserves, hormonal and metabolic shift characteristics of a switch from lipid to protein utilization (phase III, PIII) occur. These metabolic changes are paralleled by nest abandonment and stimulation of refeeding behavior. Parental behavior is then under control of two hormones with opposite effects: corticosterone (CORT) and prolactin which stimulate foraging and incubation behavior, respectively. The aim of this study was to determine the respective role of these two hormones in nest abandonment by Adélie penguins. To this end, plasma hormone levels were measured before egg-laying and at departure from the colony (i.e. when birds were relieved by their partner or abandoned their nest), and related to nutritional state and incubation success. We found that males abandoning their nest in PIII presented high CORT levels and low prolactin levels. Interestingly, males which presented high plasma levels of prolactin in PIII did not abandon. We show that although CORT is the first hormone to be affected by prolonged energy constraints, the combined effects of high CORT and low prolactin levels are necessary for parents to favor self-maintenance and abandon the nest. We provide insights into time-course changes of the endocrine profile as PIII proceeds and report that reaching proteolytic late fasting is not sufficient to induce nest abandonment in a long-lived bird. Copyright © 2010 Elsevier Inc. All rights reserved.

  7. Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.

    PubMed

    Valadares, Helder Magno Silva; Pimenta, Juliana Ramos; de Freitas, Jorge Marcelo; Duffy, Tomás; Bartholomeu, Daniella C; Oliveira, Riva de Paula; Chiari, Egler; Moreira, Maria da Consolação Vieira; Filho, Geraldo Brasileiro; Schijman, Alejandro Gabriel; Franco, Glória Regina; Machado, Carlos Renato; Pena, Sérgio Danilo Junho; Macedo, Andréa Mara

    2008-06-01

    The investigation of the importance of the genetics of Trypanosoma cruzi in determining the clinical course of Chagas disease will depend on precise characterisation of the parasites present in the tissue lesions. This can be adequately accomplished by the use of hypervariable nuclear markers such as microsatellites. However the unilocal nature of these loci and the scarcity of parasites in chronic lesions make it necessary to use high sensitivity PCR with nested primers, whose design depends on the availability of long flanking regions, a feature not hitherto available for any known T. cruzi microsatellites. Herein, making use of the extensive T. cruzi genome sequence now available and using the Tandem Repeats Finder software, it was possible to identify and characterise seven new microsatellite loci--six composed of trinucleotide (TcTAC15, TcTAT20, TcAAT8, TcATT14, TcGAG10 and TcCAA10) and one composed of tetranucleotide (TcAAAT6) motifs. All except the TcCAA10 locus were physically mapped onto distinct intergenic regions of chromosome III of the CL Brener clone contigs. The TcCAA10 locus was localised within a hypothetical protein gene in the T. cruzi genome. All microsatellites were polymorphic and useful for T. cruzi genetic variability studies. Using the TcTAC15 locus it was possible to separate the strains belonging to the T. cruzi I lineage (DTU I) from those belonging to T. cruzi II (DTU IIb), T. cruzi III (DTU IIc) and a hybrid group (DTU IId, IIe). The long flanking regions of these novel microsatellites allowed construction of nested primers and the use of full nested PCR protocols. This strategy enabled us to detect and differentiate T. cruzi strains directly in clinical specimens including heart, blood, CSF and skin tissues from patients in the acute and chronic phases of Chagas disease.

  8. Tropospheric carbon monoxide over the Pacific during HIPPO: two-way coupled simulation of GEOS-Chem and its multiple nested models

    NASA Astrophysics Data System (ADS)

    Yan, Y.-Y.; Lin, J.-T.; Kuang, Y.; Yang, D.; Zhang, L.

    2014-12-01

    Global chemical transport models (CTMs) are used extensively to study air pollution and transport at a global scale. These models are limited by coarse horizontal resolutions that do not allow for a detailed representation of small-scale nonlinear processes over the pollutant source regions. Here we couple the global GEOS-Chem CTM and its three high-resolution nested models to simulate the tropospheric carbon monoxide (CO) over the Pacific Ocean during five High-performance Instrumented Airborne Platform for Environmental Research (HIAPER) Pole-to-Pole Observations (HIPPO) campaigns between 2009 and 2011. We develop a two-way coupler, the PeKing University CouPLer (PKUCPL), allowing for the exchange and interaction of chemical constituents between the global model (at 2.5° long. × 2° lat.) and the three nested models (at 0.667° long. × 0.5° lat.) covering Asia, North America, and Europe. The coupler obtains nested model results to modify the global model simulation within the respective nested domains, and simultaneously acquires global model results to provide lateral boundary conditions (LBCs) for the nested models. Compared to the global model alone, the two-way coupled simulation results in enhanced CO concentrations in the nested domains. Sensitivity tests suggest the enhancement to be a result of improved representation of the spatial distributions of CO, nitrogen oxides, and non-methane volatile organic compounds, the meteorological dependence of natural emissions, and other resolution-dependent processes. The relatively long lifetime of CO allows for the enhancement to be accumulated and carried across the globe. We found that the two-way coupled simulation increased the global tropospheric mean CO concentrations in 2009 by 10.4%, with a greater enhancement at 13.3% in the Northern Hemisphere. Coincidently, the global tropospheric mean hydroxyl radical (OH) was reduced by 4.2%, resulting in a 4.2% enhancement in the methyl chloroform lifetime (MCF; via reaction with tropospheric OH). The resulting CO and OH contents and MCF lifetime are closer to observation-based estimates. Both the global and the two-way coupled models capture the general spatiotemporal patterns of HIPPO CO over the Pacific. The two-way coupled simulation is much closer to HIPPO CO, with a mean bias of 1.1 ppb (1.4%) below 9 km compared to the bias at -7.2 ppb (-9.2%) for the global model alone. The improvement is most apparent over the North Pacific. Our test simulations show that the global model alone could resemble the two-way coupled simulation (especially below 4 km) by increasing its global CO emissions by 15% for HIPPO-1 and HIPPO-3, by 25% for HIPPO-2 and HIPPO-4, and by 35% for HIPPO-5. This has important implications for using the global model alone to constrain CO emissions. Thus, the two-way coupled simulation is a significantly improved model tool for studying the global impacts of air pollutants from major anthropogenic source regions.

  9. Buteo Nesting Ecology: Evaluating Nesting of Swainson's Hawks in the Northern Great Plains.

    PubMed

    Inselman, Will M; Datta, Shubham; Jenks, Jonathan A; Jensen, Kent C; Grovenburg, Troy W

    2015-01-01

    Swainson's hawks (Buteo swainsoni) are long-distance migratory raptors that nest primarily in isolated trees located in areas of high grassland density. In recent years, anthropogenic conversion of grassland habitat has raised concerns about the status of the breeding population in the northern Great Plains. In 2013, we initiated a study to investigate the influence of extrinsic factors influencing Swainson's hawk nesting ecology in north-central South Dakota and south-central North Dakota. Using ground and aerial surveys, we located and monitored nesting Swainson's hawk pairs: 73 in 2013 and 120 in 2014. We documented 98 successful breeding attempts that fledged 163 chicks; 1.52 and 1.72 fledglings per successful nest in 2013 and 2014, respectively. We used Program MARK to evaluate the influence of land cover on nest survival. The top model, SDist2Farm+%Hay, indicated that nest survival (fledging at least one chick) decreased as nests were located farther from farm sites and as the percent of hay cover increased within 1200-m of the nest site (34.4%; 95% CI = 27.6%-42.3%). We used logistic regression analysis to evaluate the influence of landscape variables on nest-site selection; Swainson's hawks selected for nest sites located closer to roads. We suggest that tree belts associated with farm sites, whether occupied or not, provide critical breeding sites for Swainson's hawks. Additionally, poor breeding success may be related to the late migratory behavior of this species which requires them to occupy marginal habitat due to other raptors occupying the most suitable habitat prior to Swainson's hawks arriving to the breeding grounds.

  10. Variables associated with nest survival of Golden-winged Warblers (Vermivora chrysoptera) among vegetation communities commonly used for nesting

    USGS Publications Warehouse

    Aldinger, Kyle R.; Terhune, Theron M.; Wood, Petra Bohall; Buehler, David A.; Bakermans, Marja H.; Confer,  John L.; Flaspohler, David J.; Larkin, Jeffrey L.; Loegering, John P.; Percy, Katie L.; Roth, Amber M.; Smalling, Curtis G.

    2015-01-01

    Among shrubland- and young forest-nesting bird species in North America, Golden-winged Warblers (Vermivora chrysoptera) are one of the most rapidly declining partly because of limited nesting habitat. Creation and management of high quality vegetation communities used for nesting are needed to reduce declines. Thus, we examined whether common characteristics could be managed across much of the Golden-winged Warbler’s breeding range to increase daily survival rate (DSR) of nests. We monitored 388 nests on 62 sites throughout Minnesota, Wisconsin, New York, North Carolina, Pennsylvania, Tennessee, and West Virginia. We evaluated competing DSR models in spatial-temporal (dominant vegetation type, population segment, state, and year), intraseasonal (nest stage and time-within-season), and vegetation model suites. The best-supported DSR models among the three model suites suggested potential associations between daily survival rate of nests and state, time-within-season, percent grass and Rubus cover within 1 m of the nest, and distance to later successional forest edge. Overall, grass cover (negative association with DSR above 50%) and Rubus cover (DSR lowest at about 30%) within 1 m of the nest and distance to later successional forest edge (negative association with DSR) may represent common management targets across our states for increasing Golden-winged Warbler DSR, particularly in the Appalachian Mountains population segment. Context-specific adjustments to management strategies, such as in wetlands or areas of overlap with Blue-winged Warblers (Vermivora cyanoptera), may be necessary to increase DSR for Golden-winged Warblers.

  11. Retirement investment theory explains patterns in songbird nest-site choice

    PubMed Central

    Streby, Henry M.; Refsnider, Jeanine M.; Peterson, Sean M.; Andersen, David E.

    2014-01-01

    When opposing evolutionary selection pressures act on a behavioural trait, the result is often stabilizing selection for an intermediate optimal phenotype, with deviations from the predicted optimum attributed to tracking a moving target, development of behavioural syndromes or shifts in riskiness over an individual's lifetime. We investigated nest-site choice by female golden-winged warblers, and the selection pressures acting on that choice by two fitness components, nest success and fledgling survival. We observed strong and consistent opposing selection pressures on nest-site choice for maximizing these two fitness components, and an abrupt, within-season switch in the fitness component birds prioritize via nest-site choice, dependent on the time remaining for additional nesting attempts. We found that females consistently deviated from the predicted optimal behaviour when choosing nest sites because they can make multiple attempts at one fitness component, nest success, but only one attempt at the subsequent component, fledgling survival. Our results demonstrate a unique natural strategy for balancing opposing selection pressures to maximize total fitness. This time-dependent switch from high to low risk tolerance in nest-site choice maximizes songbird fitness in the same way a well-timed switch in human investor risk tolerance can maximize one's nest egg at retirement. Our results also provide strong evidence for the adaptive nature of songbird nest-site choice, which we suggest has been elusive primarily due to a lack of consideration for fledgling survival. PMID:24403320

  12. A simple, physiologically-based model of sea turtle remigration intervals and nesting population dynamics: Effects of temperature.

    PubMed

    Neeman, Noga; Spotila, James R; O'Connor, Michael P

    2015-09-07

    Variation in the yearly number of sea turtles nesting at rookeries can interfere with population estimates and obscure real population dynamics. Previous theoretical models suggested that this variation in nesting numbers may be driven by changes in resources at the foraging grounds. We developed a physiologically-based model that uses temperatures at foraging sites to predict foraging conditions, resource accumulation, remigration probabilities, and, ultimately, nesting numbers for a stable population of sea turtles. We used this model to explore several scenarios of temperature variation at the foraging grounds, including one-year perturbations and cyclical temperature oscillations. We found that thermally driven resource variation can indeed synchronize nesting in groups of turtles, creating cohorts, but that these cohorts tend to break down over 5-10 years unless regenerated by environmental conditions. Cohorts were broken down faster at lower temperatures. One-year perturbations of low temperature had a synchronizing effect on nesting the following year, while high temperature perturbations tended to delay nesting in a less synchronized way. Cyclical temperatures lead to cyclical responses both in nesting numbers and remigration intervals, with the amplitude and lag of the response depending on the duration of the cycle. Overall, model behavior is consistent with observations at nesting beaches. Future work should focus on refining the model to fit particular nesting populations and testing further whether or not it may be used to predict observed nesting numbers and remigration intervals. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Arboreal Ant Colonies as ‘Hot-Points’ of Cryptic Diversity for Myrmecophiles: The Weaver Ant Camponotus sp. aff. textor and Its Interaction Network with Its Associates

    PubMed Central

    Pérez-Lachaud, Gabriela; Lachaud, Jean-Paul

    2014-01-01

    Introduction Systematic surveys of macrofaunal diversity within ant colonies are lacking, particularly for ants nesting in microhabitats that are difficult to sample. Species associated with ants are generally small and rarely collected organisms, which makes them more likely to be unnoticed. We assumed that this tendency is greater for arthropod communities in microhabitats with low accessibility, such as those found in the nests of arboreal ants that may constitute a source of cryptic biodiversity. Materials and Methods We investigated the invertebrate diversity associated with an undescribed, but already threatened, Neotropical Camponotus weaver ant. As most of the common sampling methods used in studies of ant diversity are not suited for evaluating myrmecophile diversity within ant nests, we evaluated the macrofauna within ant nests through exhaustive colony sampling of three nests and examination of more than 80,000 individuals. Results We identified invertebrates from three classes belonging to 18 taxa, some of which were new to science, and recorded the first instance of the co-occurrence of two brood parasitoid wasp families attacking the same ant host colony. This diversity of ant associates corresponded to a highly complex interaction network. Agonistic interactions prevailed, but the prevalence of myrmecophiles was remarkably low. Conclusions Our data support the hypothesis of the evolution of low virulence in a variety of symbionts associated with large insect societies. Because most myrmecophiles found in this work are rare, strictly specific, and exhibit highly specialized biology, the risk of extinction for these hitherto unknown invertebrates and their natural enemies is high. The cryptic, far unappreciated diversity within arboreal ant nests in areas at high risk of habitat loss qualifies these nests as ‘hot-points’ of biodiversity that urgently require special attention as a component of conservation and management programs. PMID:24941047

  14. Reproductive plasticity and landscape heterogeneity benefit a ground-nesting bird in a fire-prone ecosystem.

    PubMed

    Carroll, J Matthew; Hovick, Torre J; Davis, Craig A; Elmore, Robert Dwayne; Fuhlendorf, Samuel D

    2017-10-01

    Disturbance is critical for the conservation of rangeland ecosystems worldwide and many of these systems are fire dependent. Although it is well established that restoring fire as an ecological process can lead to increased biodiversity in grasslands and shrublands, the underlying mechanisms driving community patterns are poorly understood for fauna in fire-prone landscapes. Much of this uncertainty stems from the paucity of studies that examine the effects of fire at scales relevant to organism life histories. We assessed the response of a non-migratory ground-dwelling bird to disturbance (i.e., prescribed fire) and environmental stochasticity over the course of a 4-yr period, which spanned years of historic drought and record rainfall. Specifically, we investigated the nesting ecology of Northern Bobwhite (Colinus virginianus; hereafter Bobwhite) to illuminate possible avenues by which individuals respond to dynamic landscape patterns during a critical reproductive stage (i.e., nesting) in a mixed-grass shrubland in western Oklahoma, USA. We found that Bobwhites exhibited extreme plasticity in nest substrate use among time since fire categories (TSF) and subsequently maintained high nest survival (e.g., 57-70%). Bobwhites were opportunistic in nest substrate use among TSF categories (i.e., 72% of nest sites in shrubs in 0-12 months post fire compared to 71% in herbaceous vegetation in >36 months post fire), yet nesting decisions were first filtered by similar structural components (i.e., vertical and horizontal cover) within the vicinity of nest sites regardless of TSF category. Despite being a non-migratory and comparatively less mobile ground-nesting bird species, Bobwhites adjusted to dynamic vegetation mosaics on a fire-prone landscape under stochastic climatic conditions that culminated in stable and high nest survival. Broadly, our findings provide a unique depiction of organism response strategies to fire at scales relevant to a critical life-stage, a topic that has been previously understudied and poorly understood. We also demonstrate how doing so can better inform conservation practices aimed at restoring fire regimes on grassland and shrubland landscapes. © 2017 by the Ecological Society of America.

  15. Nesting ecology of tundra swans on the coastal Yukon-Kuskokwim Delta, Alaska

    USGS Publications Warehouse

    Babcock, C.A.; Fowler, A.C.; Ely, Craig R.

    2002-01-01

    Nesting ecology of Tundra Swans (Cygnus columbianus columbianus) was studies the Kashunuk River near Old Chevak (61A?26a??N, 165A?27a??W), on the Yukon-Kuskokwim Delta of western Alaska from 1988-2000. Annual variation in snow-melt chronology, nesting phenology, nesting density, clutch size and nest success was examined. The same area (approximately 23 kmA?) was searched each year and nests were found as early as possible in the laying period. Laying initiation dates ranged from 1-27 May and hatch dates from 12 June a?? 4 July among pairs and years of study. The peak arrival of Tundra Swans and the phenology of nest initiation and hatch were highly correlated with the progression of ice and snow melt in spring. Nest density averaged 0.71 kmA? and 89% of nesting pairs hatched at least one egg. Incubation period ranged from 26 to 33 days with a median of 30 days. Clutch size varied significantly among years, driven by a low mean value of 3.4 eggs in 1999. Clutch sizes were generally larger than found in previous investigations on the Yukon-Kuskokwim Delta, and nearly one egg larger than reported for clutches from Alaskaa??s North Slope (=70A?N). There was no indication of reduced clutch size in years of late spring snow melt, although nesting density tended to be lower.

  16. Nesting ecology of roseate spoonbills at Nueces Bay, Texas

    USGS Publications Warehouse

    White, D.H.; Mitchell, C.A.

    1982-01-01

    We conducted a 3-year study in 1978-80 of thenesting ecology of Roseate Spoonbills (Ajaia ajaja) at Nueces Bay, Texas. Reproductive success, as well as nesting chronology, nest composition and placement, growth and development of young, egg measurements, and pollutant residues in eggs were determined. For 154 marked nests, the average clutch size was 3.0 eggs; 73% of the eggs hatched, and 87% of the nests were successful (fledged at least one young). Nest success (total fledglings: total eggs) averaged 50% during the study, or 1.5 fledglings per nest. Incubation began the day after the first egg was laid, and the incubation period for each egg was 22 days. Eggs hatched in the order that they were laid; the first and second egg hatched on consecutive days, and the second, third, and fourth egg hatched every other day. Nest composition and size were highly consistent, as was distance between nests within clusters, but nest placement varied considerably and was dependent on the vegetative configuration of the dredged-material islands. Growth rate of nestlings conformed to a 'standard' growth curve where body weight equaled that of adults at fledging. There was no difference in weight gain among siblings based on actual age. Nestlings fledged at about siz weeks of age when feather development was complete. At fledging, the bills of juveniles had almost reached adult width, but bill length was only 67% that of adults.

  17. Effect of tidal overwash on the embryonic development of leatherback turtles in French Guiana.

    PubMed

    Caut, Stéphane; Guirlet, Elodie; Girondot, Marc

    2010-05-01

    In marine turtles, the physical conditions experienced by eggs during incubation affect embryonic development. In the leatherback, hatching success is known to be low in relation to other marine turtles as a result of high embryonic mortality. Moreover, the hatching success on Yalimapo in French Guiana, one major nesting beach for this species, is lower compared to other nesting sites. We assessed the rate of leatherback turtle embryonic mortality in order to investigate the tolerance of leatherback turtle clutches laid on Yalimapo beach to tidal overwash, and we highlight causes of poor hatching success. Of the 89 nests studied, 27 were overlapped by tide at least once during the incubation period (of which five nests were lost by erosion). The hatching success was on average significantly lower in overwashed nests than in non-overwashed, highlighting the existence of embryonic developmental arrest linked to tidal inundation. The stages of developmental arrest and their proportion are linked with time, frequency and level of overwash events. In the context of global warming and associated sea-level rise, understanding the detrimental effect of tidal inundation on the development of marine turtle nests is of interest in nesting sites where turtles are likely to be forced to nest closer to the tide line, thus exposing their nests to greater risk of nest overlap with sea and tidal inundation. Copyright 2009 Elsevier Ltd. All rights reserved.

  18. Effects of three-dimensional and color patterns on nest location and progeny mortality in alfalfa leafcutting bee (Hymenoptera: Megachilidae).

    PubMed

    Guédot, Christelle; Bosch, Jordi; James, Rosalind R; Kemp, William P

    2006-06-01

    ABSTRACT In alfalfa, Medicago sativa L., seed production where high bee densities are released, alfalfa leafcutting bee, Megachile rotundata (F.) (Hymenoptera: Megachilidae), females may enter several nesting holes before locating their nests. Such levels of "wrong hole" visits lead to an increase in the time spent by females locating their own nests, thereby decreasing alfalfa pollination efficiency and possibly healthy brood production. The objectives of this study were to determine the effect of different nesting board configurations in commercial alfalfa leafcutting bee shelters (separating nesting boards, applying a three-dimensional pattern to the boards, applying a color contrast pattern, or applying a combination of three-dimensional and color contrast patterns) on nest location performance, on the incidence of chalkbrood disease, and on the incidence of broodless provisions. Separating the nesting boards inside shelters improved the ability of females to locate their nests. An increase in nest location performance also occurred in boards with the three-dimensional pattern and the combined three-dimensional and color contrast pattern, compared with the uniform board (a standard configuration currently used commercially). The percentage of provisioned cells that were broodless was not statistically different between treatments, but the percentage of larvae infected with chalkbrood decreased by half in the three-dimensional board design, compared with the uniform board.

  19. Learning Context When Studying Financial Planning in High Schools: Nesting of Student, Teacher, and Classroom Characteristics

    ERIC Educational Resources Information Center

    Danes, Sharon M.; Rodriguez, Michael C.; Brewton, Katherine E.

    2013-01-01

    Grounded in social construction theory, the current study investigates the learning context when studying financial planning in high school by analyzing the nesting of student, teacher and classroom characteristics. Key findings were that three student characteristics (initial financial knowledge, gender, senior grade level), one teacher variable…

  20. Facile synthesis of bird's nest-like TiO2 microstructure with exposed (001) facets for photocatalytic degradation of methylene blue

    NASA Astrophysics Data System (ADS)

    Zhang, Guozhong; Zhang, Shuqu; Wang, Longlu; Liu, Ran; Zeng, Yunxiong; Xia, Xinnian; Liu, Yutang; Luo, Shenglian

    2017-01-01

    The scrupulous design of hierarchical structure and highly active crystal facets exposure is essential for the creation of photocatalytic system. However, it is still a big challenge for scrupulous design of TiO2 architectures. In this paper, bird's nest-like anatase TiO2 microstructure with exposed highly active (001) surface has been successfully synthesized by a facile one-step solvothermal method. Methylene blue (MB) is chosen as a model pollutant to evaluate photocatalytic activity of as-obtained TiO2 samples. The results show that the photocatalytic activity of the bird's nest-like sample is more excellent than P25 in the degradation of MB due to high specific surface area and highly active (001) crystal facets exposure when tested under simulated solar light. Besides, it can be readily separated from the photocatalytic system by sedimentation after photocatalytic reaction, which is a significant advantage against conventional powder photocatalyst. The bird's nest-like microspheres with novel structure may have potential application in photocatalysis and other fields.

  1. The importance of floodplain forests in the conservation and management of neotropical migratory birds in the Midwest

    USGS Publications Warehouse

    Knutson, M.G.; Hoover, J.P.; Klaas, E.E.; Thompson, Frank R.

    1996-01-01

    Bottomland forests of the Central Forest Region of the Upper Midwest are found primarily on the floodplains of large rivers and include at least six types of forest communities. Birds breeding in bottomland forests are affected by extensive variation in latitude, climate, hydrology, forest succession, and change caused by anthropogenic disturbances. The floodplain forest bird community differs in species composition and in relative abundance from adjacent upland habitats. High abundances of some species are found in the floodplain and some species, such as the prothonotary warbler, brown creeper, yellow-billed cuckoo, yellow-bellied sapsucker, and great crested flycatcher, show a clear preference for floodplain forests. Studies of nesting success indicate that, for some species, nest success may be higher in the floodplain than in the uplands. Floodplain birds face threats due to large-scale loss of floodplain forest habitat. Conservation efforts should focus on restoring degraded floodplains by maintaining high tree species diversity and wide corridors. To accomplish this, the underlying hydrodynamics which support a diverse floodplain forest habitat may need to be restored. Large, contiguous tracts of floodplain and upland forests should be maintained where they exist and restored in other locations. This will provide some high quality habitat for area-sensitive neotropical migratory birds (NTMBs) in agricultural landscapes where small, scattered forest fragments are the rule. Future research efforts should examine the importance of floodplain forests in maintaining populations of neotropical migrants, especially birds experiencing population declines in adjacent uplands.

  2. Demographic consequences of nest box use for Red-footed Falcons Falco vespertinus in Central Asia

    USGS Publications Warehouse

    Bragin, Evgeny A.; Bragin, Alexander E.; Katzner, Todd

    2017-01-01

    Nest box programs are frequently implemented for the conservation of cavity-nesting birds, but their effectiveness is rarely evaluated in comparison to birds not using nest boxes. In the European Palearctic, Red-footed Falcon Falco vespertinus populations are both of high conservation concern and are strongly associated with nest box programs in heavily managed landscapes. We used a 21-year monitoring dataset collected on 753 nesting attempts by Red-footed Falcons in unmanaged natural or semi-natural habitats to provide basic information on this poorly known species; to evaluate long-term demographic trends; and to evaluate response of demographic parameters of Red-footed Falcons to environmental factors including use of nest boxes. We observed significant differences among years in laying date, offspring loss, and numbers of fledglings produced, but not in egg production. Of these four parameters, offspring loss and, to a lesser extent, number of fledglings exhibited directional trends over time. Variation in laying date and in numbers of eggs were not well explained by any one model, but instead by combinations of models, each with informative terms for nest type. Nevertheless, laying in nest boxes occurred 2.10 ± 0.70 days earlier than in natural nests. In contrast, variation in both offspring loss and numbers of fledglings produced were fairly well explained by a single model including terms for nest type, nest location, and an interaction between the two parameters (65% and 81% model weights respectively), with highest offspring loss in nest boxes on forest edges. Because, for other species, earlier laying dates are associated with more fit individuals, this interaction highlighted a possible ecological trap, whereby birds using nest boxes on forest edges lay eggs earlier but suffer greater offspring loss and produce lower numbers of fledglings than do those in other nesting settings. If nest boxes increase offspring loss for Red-footed Falcons in heavily managed landscapes where populations are at greater risk, or for the many other species of rare or endangered birds supported by nest box programs, these processes could have important demographic and conservation consequences.

  3. Nest-site selection and hatching success of waterbirds in coastal Virginia: some results of habitat manipulation

    USGS Publications Warehouse

    Rounds, R.A.; Erwin, R.M.; Portera, J.H.

    2004-01-01

    Rising sea levels in the mid-Atlantic region pose a long-term threat to marshes and their avian inhabitants. The Gull-billed Tern (Sterna nilotica), Common Tern (S. hirundo), Black Skimmer (Rynchops niger), and American Oystercatcher (Haematopus palliatus), species of concern in Virginia, nest on low shelly perimeters of salt marsh islands on the Eastern Shore of Virginia. Marsh shellpiles are free of mammalian predators, but subject to frequent floods that reduce reproductive success. In an attempt to examine nest-site selection, enhance habitat, and improve hatching success, small (2 ? 2 m) plots on five island shellpiles were experimentally elevated, and nest-site selection and hatching success were monitored from 1 May to 1 August, 2002. In addition, location, elevation, and nesting performance of all other nests in the colonies were also monitored. No species selected the elevated experimental plots preferentially over adjacent control plots at any of the sites. When all nests were considered, Common Tern nests were located significantly lower than were random point elevations at two sites, as they tended to concentrate on low-lying wrack. At two other sites, however, Common Tern nests were significantly higher than were random points. Gull-billed Terns and American Oystercatchers showed a weak preference for higher elevations on bare shell at most sites. Hatching success was not improved on elevated plots, despite the protection they provided from flooding. Because of a 7 June flood, when 47% of all nests flooded, hatching success for all species was low. Nest elevation had the strongest impact on a nest's probability of hatching, followed by nest-initiation date. Predation rates were high at small colonies, and Ruddy Turnstones (Arenaria interpres) depredated 90% of early Gull-billed Tern nests at one shellpile. The importance of nest elevation and flooding on hatching success demonstrates the potential for management of certain waterbird nesting sites. Facing threats from predators on barrier islands and rising sea levels especially in the mid-Atlantic region, several species of nesting waterbirds may benefit dramatically with modest manipulation of even small habitat patches on isolated marsh islands.

  4. Breeding behavior of northern saw-whet owls in Oregon

    USGS Publications Warehouse

    McCullough, Jenna M.; Conway, Courtney J.

    2017-01-01

    We know little about the breeding behavior of most nocturnal raptors. Nest attendance and prey delivery rates can be used as indices of relative habitat quality or extent of parental care. We used video cameras to document and observe prey delivery rates, nest attendance and bout durations at two northern saw-whet owl (Aegolius acadicus) nests in two artificial nest boxes in north-central Oregon. We collected 858 hours of video surveillance between 21 March and 01 June 2014. The number of prey deliveries per night increased as the nesting season progressed: 1.25 during laying, 1.33 during incubation, and 4.0 during the nestling phase. Prey was delivered most often between 2100 and 2200. Nest attendance by females was high during pre-laying (97.8%), laying (97.9%) and incubation (98.2%), but decreased during the nestling phase (55.7%). Nest attendance was higher during diurnal hours than nocturnal hours across all nesting phases. Duration of off bouts (recesses) was similar during pre-laying and laying (26 min), decreased during incubation (19 min), then increased during the nestling phase (55 min). One of the nesting attempts was successful and the female abandoned the other clutch 22 days after initiation. The incubation and nestling periods were 30 days each.

  5. High-temperature, high-pressure bonding of nested tubular metallic components

    DOEpatents

    Quinby, Thomas C.

    1980-01-01

    This invention is a tool for effecting high-temperature, high-compression bonding between the confronting faces of nested, tubular, metallic components. In a typical application, the tool is used to produce tubular target assemblies for irradiation in nuclear reactors or particle accelerators, the target assembly comprising a uranium foil and an aluminum-alloy substrate. The tool preferably is composed throughout of graphite. It comprises a tubular restraining member in which a mechanically expandable tubular core is mounted to form an annulus with the member. The components to be bonded are mounted in nested relation in the annulus. The expandable core is formed of individually movable, axially elongated segments whose outer faces cooperatively define a cylindrical pressing surface and whose inner faces cooperatively define two opposed, inwardly tapered, axial bores. Tapered rams extend respectively into the bores. The loaded tool is mounted in a conventional hot-press provided with evacuation means, heaters for maintaining its interior at bonding temperature, and hydraulic cylinders for maintaining a selected inwardly directed pressure on the tapered rams. With the hot-press evacuated and the loaded tool at the desired temperature, the cylinders are actuated to apply the selected pressure to the rams. The rams in turn expand the segmented core to maintain the nested components in compression against the restraining member. These conditions are maintained until the confronting faces of the nested components are joined in a continuous, uniform bond characterized by high thermal conductivity.

  6. Population ecology of breeding Pacific common eiders on the Yukon-Kuskokwim Delta, Alaska

    USGS Publications Warehouse

    Wilson, Heather M.; Flint, Paul L.; Powell, Abby N.; Grand, J. Barry; Moral, Christine L.

    2012-01-01

    Populations of Pacific common eiders (Somateria mollissima v-nigrum) on the Yukon-Kuskokwim Delta (YKD) in western Alaska declined by 50–90% from 1957 to 1992 and then stabilized at reduced numbers from the early 1990s to the present. We investigated the underlying processes affecting their population dynamics by collection and analysis of demographic data from Pacific common eiders at 3 sites on the YKD (1991–2004) for 29 site-years. We examined variation in components of reproduction, tested hypotheses about the influence of specific ecological factors on life-history variables, and investigated their relative contributions to local population dynamics. Reproductive output was low and variable, both within and among individuals, whereas apparent survival of adult females was high and relatively invariant (0.89 ± 0.005). All reproductive parameters varied across study sites and years. Clutch initiation dates ranged from 4 May to 28 June, with peak (modal) initiation occurring on 26 May. Females at an island study site consistently initiated clutches 3–5 days earlier in each year than those on 2 mainland sites. Population variance in nest initiation date was negatively related to the peak, suggesting increased synchrony in years of delayed initiation. On average, total clutch size (laid) ranged from 4.8 to 6.6 eggs, and declined with date of nest initiation. After accounting for partial predation and non-viability of eggs, average clutch size at hatch ranged from 2.0 to 5.8 eggs. Within seasons, daily survival probability (DSP) of nests was lowest during egg-laying and late-initiation dates. Estimated nest survival varied considerably across sites and years (mean = 0.55, range: 0.06–0.92), but process variance in nest survival was relatively low (0.02, CI: 0.01–0.05), indicating that most variance was likely attributed to sampling error. We found evidence that observer effects may have reduced overall nest survival by 0.0–0.36 across site-years. Study sites with lower sample sizes and more frequent visitations appeared to experience greater observer effects. In general, Pacific common eiders exhibited high spatio-temporal variance in reproductive components. Larger clutch sizes and high nest survival at early initiation dates suggested directional selection favoring early nesting. However, stochastic environmental effects may have precluded response to this apparent selection pressure. Our results suggest that females breeding early in the season have the greatest reproductive value, as these birds lay the largest clutches and have the highest probability of successfully hatching. We developed stochastic, stage-based, matrix population models that incorporated observed spatio-temporal (process) variance and co-variation in vital rates, and projected the stable stage distribution () and population growth rate (λ). We used perturbation analyses to examine the relative influence of changes in vital rates on λ and variance decomposition to assess the proportion of variation in λ explained by process variation in each vital rate. In addition to matrix-based λ, we estimated λ using capture–recapture approaches, and log-linear regression. We found the stable age distribution for Pacific common eiders was weighted heavily towards experienced adult females (≥4 yr of age), and all calculations of λ indicated that the YKD population was stable to slightly increasing (λmatrix = 1.02, CI: 1.00–1.04); λreverse-capture–recapture = 1.05, CI: 0.99–1.11; λlog-linear = 1.04, CI: 0.98–1.10). Perturbation analyses suggested the population would respond most dramatically to changes in adult female survival (relative influence of adult survival was 1.5 times that of fecundity), whereas retrospective variation in λ was primarily explained by fecundity parameters (60%), particularly duckling survival (42%). Among components of fecundity, sensitivities were highest for duckling survival, suggesti

  7. Measuring the effects of morphological changes to sea turtle nesting beaches over time with LiDAR data

    NASA Astrophysics Data System (ADS)

    Yamamoto, Kristina H.; Anderson, Sharolyn J.; Sutton, Paul C.

    2015-10-01

    Sea turtle nesting beaches in southeastern Florida were evaluated for changes from 1999 to 2005 using LiDAR datasets. Changes to beach volume were correlated with changes in several elevation-derived characteristics, such as elevation and slope. In addition, these changes to beach geomorphology were correlated to changes in nest success, illustrating that beach alterations may affect sea turtle nesting behavior. The ability to use LiDAR datasets to quickly and efficiently conduct beach comparisons for habitat use represents another benefit to this high spatial resolution data.

  8. EBV, HCMV, HHV6, and HHV7 Screening in Bone Marrow Samples from Children with Acute Lymphoblastic Leukemia

    PubMed Central

    Morales-Sánchez, A.; Pompa-Mera, E. N.; Fajardo-Gutiérrez, A.; Alvarez-Rodríguez, F. J.; Bekker-Méndez, V. C.; Flores-Chapa, J. de Diego; Flores-Lujano, J.; Jiménez-Hernández, E.; Peñaloza-González, J. G.; Rodríguez-Zepeda, M. C.; Torres-Nava, J. R.; Velázquez-Aviña, M. M.; Amador-Sánchez, R.; Alvarado-Ibarra, M.; Reyes-Zepeda, N.; Espinosa-Elizondo, R. M.; Pérez-Saldivar, M. L.; Núñez-Enríquez, J. C.; Mejía-Aranguré, J. M.; Fuentes-Pananá, E. M.

    2014-01-01

    Acute lymphoblastic leukemia (ALL) is the most common cancer in childhood worldwide and Mexico has reported one of the highest incidence rates. An infectious etiology has been suggested and supported by epidemiological evidences; however, the identity of the involved agent(s) is not known. We considered that early transmitted lymphotropic herpes viruses were good candidates, since transforming mechanisms have been described for them and some are already associated with human cancers. In this study we interrogated the direct role of EBV, HCMV, HHV6, and HHV7 human herpes viruses in childhood ALL. Viral genomes were screened in 70 bone marrow samples from ALL patients through standard and a more sensitive nested PCR. Positive samples were detected only by nested PCR indicating a low level of infection. Our result argues that viral genomes were not present in all leukemic cells, and, hence, infection most likely was not part of the initial genetic lesions leading to ALL. The high statistical power of the study suggested that these agents are not involved in the genesis of ALL in Mexican children. Additional analysis showed that detected infections or coinfections were not associated with prognosis. PMID:25309913

  9. EBV, HCMV, HHV6, and HHV7 screening in bone marrow samples from children with acute lymphoblastic leukemia.

    PubMed

    Morales-Sánchez, A; Pompa-Mera, E N; Fajardo-Gutiérrez, A; Alvarez-Rodríguez, F J; Bekker-Méndez, V C; Flores-Chapa, J de Diego; Flores-Lujano, J; Jiménez-Hernández, E; Peñaloza-González, J G; Rodríguez-Zepeda, M C; Torres-Nava, J R; Velázquez-Aviña, M M; Amador-Sánchez, R; Alvarado-Ibarra, M; Reyes-Zepeda, N; Espinosa-Elizondo, R M; Pérez-Saldivar, M L; Núñez-Enríquez, J C; Mejía-Aranguré, J M; Fuentes-Pananá, E M

    2014-01-01

    Acute lymphoblastic leukemia (ALL) is the most common cancer in childhood worldwide and Mexico has reported one of the highest incidence rates. An infectious etiology has been suggested and supported by epidemiological evidences; however, the identity of the involved agent(s) is not known. We considered that early transmitted lymphotropic herpes viruses were good candidates, since transforming mechanisms have been described for them and some are already associated with human cancers. In this study we interrogated the direct role of EBV, HCMV, HHV6, and HHV7 human herpes viruses in childhood ALL. Viral genomes were screened in 70 bone marrow samples from ALL patients through standard and a more sensitive nested PCR. Positive samples were detected only by nested PCR indicating a low level of infection. Our result argues that viral genomes were not present in all leukemic cells, and, hence, infection most likely was not part of the initial genetic lesions leading to ALL. The high statistical power of the study suggested that these agents are not involved in the genesis of ALL in Mexican children. Additional analysis showed that detected infections or coinfections were not associated with prognosis.

  10. Absence of Asymptomatic Malaria Infection in a Cross-sectional Study in Iranshahr District, Iran under Elimination Programmes

    PubMed Central

    PIRAHMADI, Sakineh; ZAKERI, Sedigheh; RAEISI, Ahmad

    2017-01-01

    Background: Asymptomatic malaria infection provides a reservoir of parasites, causing the persistence of malaria transmission. It accounts an important challenge for successful management of the control, elimination, and eradication programmes in any malaria-endemic region. This investigation was designed to assess the presence and the prevalence of asymptomatic carriers in Iranshahr district of Sistan and Baluchistan Province (2013–2014), with a considerable population movement, during the malaria elimination phase in Iran. Methods: Finger-prick blood samples were collected from symptomless (n=250) and febrile (n=50) individuals residing in Iranshahr district, easthern Iran (Hoodian, Mand, Chah-e Giji, Jolgehashem, Esfand, Dalgan and Chahshour) during Jan 2013 to Dec 2014, and Plasmodium infections were detected using light microscopic and highly sensitive nested-PCR techniques. Results: Thick and thin Giemsa-stained blood smears were negative for Plasmodium parasites. In addition, based on nested-PCR analysis, no P. vivax, P. falciparum, and P. malariae parasites were detected among the studied individuals. Conclusion: Investigation the absence of asymptomatic carriers in Iranshahr district was illustrated and achieving malaria elimination in this area is feasible in a near future. PMID:28761465

  11. A Novel Isothermal Assay of Borrelia burgdorferi by Recombinase Polymerase Amplification with Lateral Flow Detection.

    PubMed

    Liu, Wei; Liu, Hui-Xin; Zhang, Lin; Hou, Xue-Xia; Wan, Kang-Lin; Hao, Qin

    2016-08-03

    A novel isothermal detection for recombinase polymerase amplification with lateral flow (LF-RPA) was established for Borrelia burgdorferi (B. burgdorferi) detection in this study. This assay with high sensitivity and specificity can get a visible result without any additional equipment in 30 min. We designed a pair of primers according to recA gene of B. burgdorferi strains and a methodology evaluation was performed. The results showed that the RPA assay based on the recA gene was successfully applied in B. burgdorferi detection, and its specific amplification was only achieved from the genomic DNA of B. burgdorferi. The detection limit of the new assay was about 25 copies of the B. burgdorferi genomic DNA. Twenty Lyme borreliosis patients' serum samples were detected by LF-RPA assay, real-time qPCR and nested-PCR. Results showed the LF-RPA assay is more effective than nested-PCR for its shorter reaction time and considerably higher detection rate. This method is of great value in clinical rapid detection for Lyme borreliosis. Using the RPA assay might be a megatrend for DNA detection in clinics and endemic regions.

  12. Probabilistic methods for sensitivity analysis and calibration in the NASA challenge problem

    DOE PAGES

    Safta, Cosmin; Sargsyan, Khachik; Najm, Habib N.; ...

    2015-01-01

    In this study, a series of algorithms are proposed to address the problems in the NASA Langley Research Center Multidisciplinary Uncertainty Quantification Challenge. A Bayesian approach is employed to characterize and calibrate the epistemic parameters based on the available data, whereas a variance-based global sensitivity analysis is used to rank the epistemic and aleatory model parameters. A nested sampling of the aleatory–epistemic space is proposed to propagate uncertainties from model parameters to output quantities of interest.

  13. Probabilistic methods for sensitivity analysis and calibration in the NASA challenge problem

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Safta, Cosmin; Sargsyan, Khachik; Najm, Habib N.

    In this study, a series of algorithms are proposed to address the problems in the NASA Langley Research Center Multidisciplinary Uncertainty Quantification Challenge. A Bayesian approach is employed to characterize and calibrate the epistemic parameters based on the available data, whereas a variance-based global sensitivity analysis is used to rank the epistemic and aleatory model parameters. A nested sampling of the aleatory–epistemic space is proposed to propagate uncertainties from model parameters to output quantities of interest.

  14. Rapid detection of 21-hydroxylase deficiency mutations by allele-specific in vitro amplification and capillary zone electrophoresis.

    PubMed

    Carrera, P; Barbieri, A M; Ferrari, M; Righetti, P G; Perego, M; Gelfi, C

    1997-11-01

    A quick diagnosis of the classic form of 21-hydroxylase deficiency (simple virilizing and salt wasting) is of great importance, especially for prenatal diagnosis and treatment in pregnancies at risk. A method for simultaneous detection of common point mutations in the P450c21 B gene is here proposed by combining a nested PCR amplification refractory mutation system (ARMS) with capillary zone electrophoresis (CZE) in sieving liquid polymers. In the first PCR, B genes are selectively amplified. In the nested reaction, ARMS-detected wild-type and mutated alleles are separately pooled and resolved by CZE. CZE is performed in coated capillaries in the presence of 30 g/L hydroxyethyl cellulose in the background electrolyte for size separation of the DNA analytes. For high-sensitivity detection the electrophoresis buffer contains the fluorescent dye SYBR Green I. Laser-induced fluorescence detection is obtained by excitation at 488 nm and signal collection at 520 nm. Specificity and reproducibility of the protocols were established by using samples from 75 Italian families with 21-hydroxylase deficiency already genotyped by allele-specific oligonucleotide hybridization or direct sequencing. Whereas dot-blot is time consuming because of the high number of hybridizations with radioactive probes, this present protocol is more rapid, giving sufficient separation on CZE after PCR reactions without preconcentration or desalting of samples.

  15. A two-way nested simulation of the oceanic circulation in the Southwestern Atlantic

    NASA Astrophysics Data System (ADS)

    Combes, Vincent; Matano, Ricardo P.

    2014-02-01

    This article presents the results of a high-resolution (1/12°), two-way nested simulation of the oceanic circulation in the southwestern Atlantic region. A comparison between the model results and extant observations indicates that the nested model has skill in reproducing the best-known aspects of the regional circulation, e.g., the volume transport of the ACC, the latitudinal position of the BMC, the shelf break upwelling of Patagonia, and the Zapiola Anticyclone. Sensitivity experiments indicate that the bottom stress parameterization significantly impacts the mean location of the Brazil/Malvinas Confluence and the transport of the Zapiola Anticyclone. The transport of the Brazil Current strengthens during the austral summer and weakens during the austral winter. These variations are driven by the wind stress curl over the southwestern Atlantic. The variations of the transport of the Malvinas Current are out of phase with those of the Brazil Current. Most of the seasonal variability of this current is concentrated in the offshore portion of the jet, the inshore portion has a weak seasonality that modulates the magnitude of the Patagonian shelf break upwelling. Using passive tracers we show that most of the entrainment of deep waters into the shelf occurs in the southernmost portion of the Patagonian shelf and along the inshore boundary of the Brazil Current. Shelf waters are preferentially detrained near the Brazil/Malvinas Confluence. Consistent with previous studies, our simulation also shows that south of ˜42°S the Malvinas Current is composed of two jets, which merge near 42°S to form a single jet farther north.

  16. Specific detection of Xanthomonas axonopodis pv. dieffenbachiae in anthurium (Anthurium andreanum) tissues by nested PCR.

    PubMed

    Robène-Soustrade, Isabelle; Laurent, Philippe; Gagnevin, Lionel; Jouen, Emmanuel; Pruvost, Olivier

    2006-02-01

    Efficient control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires a sensitive and reliable diagnostic tool. A nested PCR test was developed from a sequence-characterized amplified region marker identified by randomly amplified polymorphic DNA PCR for the detection of X. axonopodis pv. dieffenbachiae. Serological and pathogenicity tests were performed concurrently with the nested PCR test with a large collection of X. axonopodis pv. dieffenbachiae strains that were isolated worldwide and are pathogenic to anthurium and/or other aroids. The internal primer pair directed amplification of the expected product (785 bp) for all 70 X. axonopodis pv. dieffenbachiae strains pathogenic to anthurium tested and for isolates originating from syngonium and not pathogenic to anthurium. This finding is consistent with previous studies which indicated that there is a high level of relatedness between strains from anthurium and strains from syngonium. Strains originating from the two host genera can be distinguished by restriction analysis of the amplification product. No amplification product was obtained with 98 strains of unrelated phytopathogenic bacteria or saprophytic bacteria from the anthurium phyllosphere, except for a weak signal obtained for one X. axonopodis pv. allii strain. Nevertheless, restriction enzyme analysis permitted the two pathovars to be distinguished. The detection threshold obtained with pure cultures or plant extracts (10(3) CFU ml(-1)) allowed detection of the pathogen from symptomless contaminated plants. This test could be a useful diagnostic tool for screening propagation stock plant material and for monitoring international movement of X. axonopodis pv. dieffenbachiae.

  17. Analysis of Wading Bird use of Impounded Wetland Habitat on the Kennedy Space Center/Merritt Island National Wildlife Refuge, 1987-1998

    NASA Technical Reports Server (NTRS)

    Stolen, Eric D.; Breininger, David R.; Smith, Rebecca B.; Quincy, Charlie (Technical Monitor)

    2002-01-01

    This report summarizes results of the first eleven years of monthly aerial surveys of wading bird use of foraging habitats within impoundments on the Kennedy Space Center/Merritt Island National Wildlife Refuge. Some impoundments were used much more heavily by wading birds than were others. Analysis suggests that an increase in interspersion of open water and vegetated habitats is preferred foraging habitat. Many wading bird species increased their use of vegetated habitat in Fall and Winter when impoundments were flooded. The mean number of wading birds per survey was greatest during the Pre-nesting and Nesting seasons, declined during Post-nesting season, and was lowest during Winter when water levels within impoundments were high. During these times, shallow habitat along the IRL shoreline provided alternative habitats for wading birds. Various measures of monthly precipitation and impoundment water level were well correlated with the numbers of wading birds observed. Numbers of nesting attempts was steady during the study period, with the exception of an unusually high number of attempts in 1990. White Ibis accounted for over half of all wading bird nests counted. The mean number of nests per colony decreased during the study period, and the number of individual colonies increased.

  18. Seasonal patterns of prey availability and the foraging behavior of arctic foxes (Alopex lagopus) in a waterfowl nesting area

    USGS Publications Warehouse

    Stickney, Alice

    1991-01-01

    The foraging behavior of arctic foxes was observed in a waterfowl nesting area on the Yukon–Kuskokwim Delta of Alaska in 1985–1986. Observations were made during peak fox activity from two towers, 3 m high, located in different community types. Data were collected continuously for individual foxes on specific activities, the community in which activities occurred, and the type of food obtained. After migratory birds started nesting in the area, the food potentially available to foxes changed from microtines, old caches, and carrion to include eggs and birds. This change was reflected in the foraging behavior of the foxes as they switched to predation on eggs. After nesting began, the search success rate of foxes increased (from <30% to >50%) and search duration decreased (mean 19.7 s before nest initiation versus mean 9.4 s in mid-incubation) as the rate of food acquisition increased. Over 80% of the eggs taken by foxes were cached rather than eaten immediately, which extended the availability of this temporally limited resource to foxes. Eggs were the primary prey of arctic foxes during the nesting stages in both years, even though microtine populations were high in one year (1985) and low in the other (1986).

  19. Nest predation risk explains variation in avian clutch size

    USGS Publications Warehouse

    Dillon, Kristen G.; Conway, Courtney J.

    2018-01-01

    Questions about the ecological drivers of, and mechanistic constraints on, productivity have driven research on life-history evolution for decades. Resource availability and offspring mortality are considered among the 2 most important influences on the number of offspring per reproductive attempt. We used a factorial experimental design to manipulate food abundance and perceived offspring predation risk in a wild avian population (red-faced warblers; Cardellina rubrifrons) to identify the mechanistic cause of variation in avian clutch size. Additionally, we tested whether female quality helped explain the extant variation in clutch size. We found no support for the Food Limitation or Female Quality Hypotheses, but we did find support for both predictions of the Nest Predation Risk Hypothesis. Females that experienced an experimentally heightened perception of offspring predation risk responded by laying a smaller clutch than females in the control group. Additionally, predation rates at artificial nests were highest where red-faced warbler clutch size was smallest (at high elevations). Life-history theory predicts that an individual should invest less in reproduction when high nest predation risk reduces the likely benefit from that nesting attempt and, indeed, we found that birds exhibit phenotypic plasticity in clutch size by laying fewer eggs in response to increasing nest predation risk.

  20. Colony Fusion in a Parthenogenetic Ant, Pristomyrmex punctatus

    PubMed Central

    Satow, Show; Satoh, Toshiyuki; Hirota, Tadao

    2013-01-01

    In the ant Pristomyrmex punctatus Smith (Hymenoptera: Formicidae), all young workers lay a small number of eggs parthenogenetically. Some colonies consist of monoclonal individuals that provide high inclusive fitness, according to the kin selection theory. However, in some populations, a majority of the colonies contain multiple lineages. Intracolonial genetic variation of parthenogenetic ants cannot be explained by the multiple mating of single founderesses or by the foundation of a colony by multiple foundresses, which are the usual causes of genetically diverse colonies in social insects. Here, we hypothesized that the fusion of established colonies might facilitate the formation of multiclonal colonies. Colony fusion decreases indirect benefits because of the reduction in intracolonial relatedness. However, when suitable nesting places for overwintering are scarce, colony fusion provides a strategy for the survival of colonies. Here, ants derived from different colonies were allowed to encounter one another in a container with just one nesting place. Initially, high aggression was observed; however, after several days, no aggression was observed and the ants shared the nest. When the fused colonies were allowed to transfer to two alternative nests, ants from different colonies occupied the same nest. This study highlights the importance of limiting the number of nesting places in order to understand the genetic diversity of parthenogenetic ant colonies. PMID:23895053

  1. Factors affecting chick provisioning by Caspian Terns nesting in the Columbia River estuary

    USGS Publications Warehouse

    Anderson, Scott K.; Roby, D.D.; Lyons, Donald E.; Collis, K.

    2005-01-01

    We investigated factors affecting chick provisioning by radio-tagged Caspian Terns (Sterna caspia) nesting in a large colony on East Sand Island in the Columbia River estuary during 2001. Caspian Tern predation on juvenile salmonids (Oncorhynchus spp.) in the estuary prompted resource managers to relocate ca. 9,000 pairs of terns nesting on Rice Island (river km 34) to East Sand Island (river km 8), where terns were expected to consume fewer salmonids in favor of marine forage fishes. This study investigated factors influencing foraging success, diet composition, and overall reproductive success at the managed Caspian Tern colony. Our results indicated that daytime colony attendance by nesting terns averaged 64% and decreased throughout the chick-rearing period, while duration of foraging trips averaged 47 min and increased during the same period; these seasonal changes were more strongly related to date than chick age. Average meal delivery rates to 2-chick broods (0.88 meals h-1) were 2.6 times greater than to 1-chick broods (0.33 meals h-1). Parents delivered more juvenile salmonids to chicks during ebb tides than flood tides, but meal delivery rates to the nest remained constant, suggesting diet composition tracks relative availability of prey species. Foraging trips resulting in delivery of juvenile salmonids averaged 68% longer than foraging trips for schooling marine forage fishes, indicating higher availability of marine prey near the colony. High availability of marine forage fish in the Columbia River estuary during 2001 was apparently responsible for high colony attendance, short foraging trips, high chick meal delivery rates, and high nesting success of Caspian Terns on East Sand Island.

  2. Constraints on productivity of wild Nene or Hawaiian geese Branta sandvicensis

    USGS Publications Warehouse

    Banko, P.C.

    1992-01-01

    I investigated constraints on the productivity of wild Nene on Hawaii and Maui during 1978-81. These populations were composed largely of captive-reared birds. Recruitment of young was low. Of 140 breeding attempts, 36% resulted in successful nests and 7% produced fledglings. Annual productivity was limited because: 1) relatively few available pairs attempted to breed (58% on Hawaii; 46% on Maui), 2) average rate of nest success was low (44%), and 3) gosling survival was low <39%). Low incidence of nesting suggests that many females could not accumulate sufficient body reserves for egg-laying and incubation due to poor foraging conditions or poorly developed foraging skills. Nest failure was high due to predation on eggs and incubating females by the introduced mongoose. Gosling mortality was high because of poor foraging conditions near many nests, forcing broods to travel over rugged, volcanic terrain to distant rearing areas. In addition, some goslings were killed by predators. Nene populations would benefit most from improved foraging opportunities for adult females and goslings and from reduced predator populations.

  3. Evaluation of Different PCR-Based Assays and LAMP Method for Rapid Detection of Phytophthora infestans by Targeting the Ypt1 Gene

    PubMed Central

    Khan, Mehran; Li, Benjin; Jiang, Yue; Weng, Qiyong; Chen, Qinghe

    2017-01-01

    Late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases affecting potato and tomato worldwide. Early diagnosis of the P. infestans pathogen causing late blight should be the top priority for addressing disease epidemics and management. In this study, we performed a loop-mediated isothermal amplification (LAMP) assay, conventional polymerase chain reaction (PCR), nested PCR, and real-time PCR to verify and compare the sensitivity and specificity of the reaction based on the Ypt1 (Ras-related protein) gene of P. infestans. In comparison with the PCR-based assays, the LAMP technique led to higher specificity and sensitivity, using uncomplicated equipment with an equivalent time frame. All 43 P. infestans isolates, yielded positive detection results using LAMP assay showing no cross reaction with other Phytophthora spp., oomycetes or fungal pathogens. The LAMP assay yielded the lowest detectable DNA concentration (1.28 × 10-4 ng μL-1), being 10 times more sensitive than nested PCR (1.28 × 10-3 ng μL-1), 100 times more sensitive than real-time PCR (1.28 × 10-2 ng μL-1) and 103 times more sensitive than the conventional PCR assay (1.28 × 10-1 ng μL-1). In the field experiment, the LAMP assay outperformed the other tests by amplifying only diseased tissues (leaf and stem), and showing no positive reaction in healthy tissues. Overall, the LAMP assay developed in this study provides a specific, sensitive, simple, and effective visual method for detection of the P. infestans pathogen, and is therefore suitable for application in early prediction of the disease to reduce the risk of epidemics. PMID:29051751

  4. Evaluation of Different PCR-Based Assays and LAMP Method for Rapid Detection of Phytophthora infestans by Targeting the Ypt1 Gene.

    PubMed

    Khan, Mehran; Li, Benjin; Jiang, Yue; Weng, Qiyong; Chen, Qinghe

    2017-01-01

    Late blight, caused by the oomycete Phytophthora infestans , is one of the most devastating diseases affecting potato and tomato worldwide. Early diagnosis of the P. infestans pathogen causing late blight should be the top priority for addressing disease epidemics and management. In this study, we performed a loop-mediated isothermal amplification (LAMP) assay, conventional polymerase chain reaction (PCR), nested PCR, and real-time PCR to verify and compare the sensitivity and specificity of the reaction based on the Ypt1 (Ras-related protein) gene of P. infestans. In comparison with the PCR-based assays, the LAMP technique led to higher specificity and sensitivity, using uncomplicated equipment with an equivalent time frame. All 43 P. infestans isolates, yielded positive detection results using LAMP assay showing no cross reaction with other Phytophthora spp., oomycetes or fungal pathogens. The LAMP assay yielded the lowest detectable DNA concentration (1.28 × 10 -4 ng μL -1 ), being 10 times more sensitive than nested PCR (1.28 × 10 -3 ng μL -1 ), 100 times more sensitive than real-time PCR (1.28 × 10 -2 ng μL -1 ) and 10 3 times more sensitive than the conventional PCR assay (1.28 × 10 -1 ng μL -1 ). In the field experiment, the LAMP assay outperformed the other tests by amplifying only diseased tissues (leaf and stem), and showing no positive reaction in healthy tissues. Overall, the LAMP assay developed in this study provides a specific, sensitive, simple, and effective visual method for detection of the P. infestans pathogen, and is therefore suitable for application in early prediction of the disease to reduce the risk of epidemics.

  5. Nesting ecology and nest survival of lesser prairie-chickens on the Southern High Plains of Texas

    USGS Publications Warehouse

    Grisham, Blake A.; Borsdorf, Philip K.; Boal, Clint W.; Boydston, Kathy K.

    2014-01-01

    The decline in population and range of lesser prairie-chickens (Tympanuchus pallidicinctus) throughout the central and southern Great Plains has raised concerns considering their candidate status under the United States Endangered Species Act. Baseline ecological data for lesser prairie-chickens are limited, especially for the shinnery oak-grassland communities of Texas. This information is imperative because lesser prairie-chickens in shinnery oak grasslands occur at the extreme southwestern edge of their distribution. This geographic region is characterized by hot, arid climates, less fragmentation, and less anthropogenic development than within the remaining core distribution of the species. Thus, large expanses of open rangeland with less anthropogenic development and a climate that is classified as extreme for ground nesting birds may subsequently influence nest ecology, nest survival, and nest site selection differently compared to the rest of the distribution of the species. We investigated the nesting ecology of 50 radio-tagged lesser prairie-chicken hens from 2008 to 2011 in the shinnery oak-grassland communities in west Texas and found a substantial amount of inter-annual variation in incubation start date and percent of females incubating nests. Prairie-chickens were less likely to nest near unimproved roads and utility poles and in areas with more bare ground and litter. In contrast, hens selected areas dominated by grasses and shrubs and close to stock tanks to nest. Candidate models including visual obstruction best explained daily nest survival; a 5% increase in visual obstruction improved nest survival probability by 10%. The model-averaged probability of a nest surviving the incubation period was 0.43 (SE = 0.006; 95% CI: 0.23, 0.56). Our findings indicate that lesser prairie-chicken reproduction during our study period was dynamic and was correlated with seasonal weather patterns that ultimately promoted greater grass growth earlier in the nesting season that provided visual obstruction from predators.

  6. The influence of coyotes on an urban Canada goose population in the Chicago metropolitan area

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Brown, Justin L.; /Ohio State U.

    Canada geese (Branta canadensis) have become common in many urban areas, often creating nuisance problems for human residents. The presence of urban geese has raised concerns about the spread of disease, increased erosion, excessive noise, eutrophication of waterways, and general nuisance problems. Goose populations have grown due to an increase in urbanization resulting in an abundance of high quality food (urban grass) and suitable nesting sites, as well as a decrease in some predators. I monitored nest predation in the Chicago suburbs during the 2004 and 2005 nesting seasons using 3 nest monitoring techniques to identify predators: video cameras, plasticinemore » eggs, and sign from nest using a classification tree analysis. Of 58 nests monitored in 2004 and 286 in 2005, only raccoons (Procyon lotor) and coyotes (Canis latrans) were identified as nest predators. Raccoons were responsible for 22-25% of depredated nests, but were rarely capable of depredating nests that were actively defended by a goose. Coyotes were responsible for 75-78% of all Canada goose nest depredation and were documented killing one adult goose and feeding on several others. The coyote is a top-level predator that had increased in many metropolitan areas in recent years. To determine if coyotes were actively hunting geese or eggs during the nesting season, I analyzed coyote habitat selection between nesting and pre-nesting or post-nesting seasons. Coyote home ranges (95% Minimum Convex Polygon) were calculated for 19 coyotes to examine third order habitat selection related to goose nest abundance. A 100 m buffer (buffer habitat) was created and centered on each waterway edge and contained 90% of all nests. Coyotes showed selection for habitats during all seasons. Buffer habitat was the top ranked habitat in both pre-nesting and nesting seasons, but dropped to third ranked in post-nesting season. Habitat selection across seasons was compared using a repeated measures MANOVA. Habitat selection between pre-nesting and nesting seasons (P=0.72) were similar, while between post-nesting and nesting seasons there was a nearly significant difference (P=0.07). The insignificant change in habitat use across seasons suggests that coyotes did not switch habitat use to take advantage of goose nests. Alternatively, the change in ranking of buffer habitat across seasons suggests that coyotes may have switched habitat use to take advantage of goose nests. The results are not clear as large individual variation between coyotes due to differences in habitat availability, and social status interfere with the results of the analysis. Even though I failed to find strong support for coyotes actively hunting goose nests, they nevertheless were the primary nest predator in the area and may influence Canada goose populations. To determine the potential influence of coyotes on the Canada goose population, I created a Canada goose matrix population model that included variables such as coyote predation on adults and nests as well as coyote influence on nest desertion. Using the base population model I calculated the Canada goose population to be increasing with {lambda} = 1.055. The removal of all coyote influence on the goose population would allow {lambda} to increase to 1.214. Nest predation was the most important factor related to coyotes: the removal of coyote nest predation from the model resulted in a population growth rate {lambda} = 1.157. Modeling results suggest coyotes are serving as a limiting factor for the Canada goose population within the Chicago metropolitan area.« less

  7. Risk of predation and weather events affect nest site selection by sympatric Pacific (Gavia pacifica) and Yellow-billed (Gavia adamsii) loons in Arctic habitats

    USGS Publications Warehouse

    Haynes, Trevor B.; Schmutz, Joel A.; Lindberg, Mark S.; Rosenberger, Amanda E.

    2014-01-01

    Pacific (Gavia pacifica) and Yellow-billed (G. adamsii) loons nest sympatrically in Arctic regions. These related species likely face similar constraints and requirements for nesting success; therefore, use of similar habitats and direct competition for nesting habitat is likely. Both of these loon species must select a breeding lake that provides suitable habitat for nesting and raising chicks; however, characteristics of nest site selection by either species on interior Arctic lakes remains poorly understood. Here, logistic regression was used to compare structural and habitat characteristics of all loon nest locations with random points from lakes on the interior Arctic Coastal Plain, Alaska. Results suggest that both loon species select nest sites to avoid predation and exposure to waves and shifting ice. Loon nest sites were more likely to be on islands and peninsulas (odds ratio = 16.13, 95% CI = 4.64–56.16) than mainland shoreline, which may help loons avoid terrestrial predators. Further, nest sites had a higher degree of visibility (mean degrees of visibility to 100 and 200 m) of approaching predators than random points (odds ratio = 2.57, 95% CI = 1.22–5.39). Nests were sheltered from exposure, having lower odds of being exposed to prevailing winds (odds ratio = 0.34, 95% CI = 0.13–0.92) and lower odds of having high fetch values (odds ratio = 0.46, 95% CI = 0.22–0.96). Differences between Pacific and Yellow-billed loon nesting sites were subtle, suggesting that both species have similar general nest site requirements. However, Yellow-billed Loons nested at slightly higher elevations and were more likely to nest on peninsulas than Pacific Loons. Pacific Loons constructed built up nests from mud and vegetation, potentially in response to limited access to suitable shoreline due to other territorial loons. Results suggest that land managers wishing to protect habitats for these species should focus on lakes with islands as well as shorelines sheltered from exposure to prevailing wind and ice patterns.

  8. Focusing X-Ray Telescopes

    NASA Technical Reports Server (NTRS)

    O'Dell, Stephen; Brissenden, Roger; Davis, William; Elsner, Ronald; Elvis, Martin; Freeman, Mark; Gaetz, Terrance; Gorenstein, Paul; Gubarev, Mikhall; Jerlus, Diab; hide

    2010-01-01

    During the half-century history of x-ray astronomy, focusing x-ray telescopes, through increased effective area and finer angular resolution, have improved sensitivity by 8 orders of magnitude. Here, we review previous and current x-ray-telescope missions. Next, we describe the planned next-generation x-ray-astronomy facility, the International X-ray Observatory (IXO). We conclude with an overview of a concept for the next next-generation facility, Generation X. Its scientific objectives will require very large areas (about 10,000 sq m) of highly-nested, lightweight grazing-incidence mirrors, with exceptional (about 0.1-arcsec) resolution. Achieving this angular resolution with lightweight mirrors will likely require on-orbit adjustment of alignment and figure.

  9. [Evaluation of the usefulness of various PCR method variations and nucleic acid hybridization for CMV infection in immunosuppressed patients].

    PubMed

    Siennicka, J; Trzcińska, A; Litwińska, B; Durlik, M; Seferyńska, I; Pałynyczko, G; Kańtoch, M

    2000-01-01

    In diagnosis of CMV infection various laboratory methods are used. The methods based on detection of viral nucleic acids have been introduced routinely in many laboratories. The aim of this study was to compare nucleic acid hybridisation method and various variants of PCR methods with respect to their ability to detect CMV DNA. The studied material comprised 60 blood samples from 19 patients including 13 renal transplant recipients and 6 with acute leukaemia. The samples were subjected to hybridisation (Murex Hybrid Capture System CMV DNA) and PCR carried out in 3 variants: with one pair of primers (single PCR), nested PCR and Digene SHARP System with detection of PCR product using a genetic probe in ELISA system. The sensitivity of the variants ranged from 10(0) particles of viral DNA in nested PCR to 10(2) in single PCR. The producer claimed the sensitivity of the hybridisation test to be 3 x 10(5) and it seems to be sufficient for detection of CMV infection. The obtained results show that sensitivity of hybridisation was comparable to that of single PCR and the possibility of obtaining quantitative results makes it superior, on efficacy of antiviral therapy, especially in monitoring CMV infection in immunossuppressed patients and in following the efficacy of antiviral treatment.

  10. Breeding productivity and survival of the endangered Hawai'i Creeper in a wet forest refuge on Mauna Kea, Hawai'i

    USGS Publications Warehouse

    Collins, Mark

    2001-01-01

    We studied the demography of the endangered Hawai‘i Creeper (Oreomystis muna) from 1994-1999 at three sites in Hakalau Forest National Wildlife Refuge (NWR). Hawai‘i Creepers bred from January to June, with peak breeding in February through May (about 120-l 80 days), and molted from May to August. A small proportion (4.9%) of individuals overlapped breeding and molting activities. We located and monitored the fates of 60 nests. Mean clutch size was 2.1 eggs, nest building required 19 days, incubation was 16 to 17 days, and nestling period lasted 18 days. Of all nest attempts, 25% were abandoned before egg laying, 6.7% were removed for captive propagation, 13.3% had undetermined fates, 38.3% failed during incubation or nestling periods, and 16.7% were successful. Thus, of 33 nests that were active through egg laying and outcome was confirmed, only 30% were successful. The daily survival rate of active nests was 0.960 t 0.009 SE. An average of 1.7 chicks fledged from successful nests. Thirty-two percent of hatch-year birds were alive and in the study area at least one year later. Annual adult survival was high (0.88 t 0.03). The primary factors limiting productivity of Hawai‘i Creeper in Hakalau Forest NWR appear to be low reproductive potential in combination with high rates of nesting failure. Further research into the causes of nest failure, the length of the breeding season, and renesting behavior of females is needed, and protection of the forest from the degrading impacts of introduced mammals is paramount.

  11. Multi-scale habitat selection in highly territorial bird species: Exploring the contribution of nest, territory and landscape levels to site choice in breeding rallids (Aves: Rallidae)

    NASA Astrophysics Data System (ADS)

    Jedlikowski, Jan; Chibowski, Piotr; Karasek, Tomasz; Brambilla, Mattia

    2016-05-01

    Habitat selection often involves choices made at different spatial scales, but the underlying mechanisms are still poorly understood, and studies that investigate the relative importance of individual scales are rare. We investigated the effect of three spatial scales (landscape, territory, nest-site) on the occurrence pattern of little crake Zapornia parva and water rail Rallus aquaticus at 74 ponds in the Masurian Lakeland, Poland. Habitat structure, food abundance and water chemical parameters were measured at nests and random points within landscape plots (from 300-m to 50-m radius), territory (14-m) and nest-site plots (3-m). Regression analyses suggested that the most relevant scale was territory level, followed by landscape, and finally by nest-site for both species. Variation partitioning confirmed this pattern for water rail, but also highlighted the importance of nest-site (the level explaining the highest share of unique variation) for little crake. The most important variables determining the occurrence of both species were water body fragmentation (landscape), vegetation density (territory) and water depth (at territory level for little crake, and at nest-site level for water rail). Finally, for both species multi-scale models including factors from different levels were more parsimonious than single-scale ones, i.e. habitat selection was likely a multi-scale process. The importance of particular spatial scales seemed more related to life-history traits than to the extent of the scales considered. In the case of our study species, the territory level was highly important likely because both rallids have to obtain all the resources they need (nest site, food and mates) in relatively small areas, the multi-purpose territories they defend.

  12. Gaseous templates in ant nests.

    PubMed

    Cox, M D; Blanchard, G B

    2000-05-21

    We apply a diffusion model to the atmosphere of ant nests. With particular reference to carbon dioxide (CO2), we explore analytically and numerically the spatial and temporal patterns of brood- or worker-produced gases in nests. The maximum concentration within a typical one-chamber ant nest with approximately 200 ants can reach 12.5 times atmospheric concentration, reaching 95% of equilibrium concentrations within 15 min. Maximum concentration increases with increasing number of ants in the nest (or production rate of the gas), distance between the centre of the nest ants and the nest entrance, entrance length, wall thickness, and with decreasing entrance width, wall permeability and diffusion coefficient. The nest can be divided into three qualitatively distinct regions according to the shape of the gradient: a plateau of high concentration in the back half of the nest; an intermediate region of increasingly steep gradient towards the entrance; and a steep linear gradient in the entrance tunnel. These regions are robust to changes in gas concentrations, but vary with changes in nest architecture. The pattern of diffusing gases contains information about position and orientation relative to gas sources and sinks, and about colony state, including colony size, activity state and aspects of nest architecture. We discuss how this diffusion pattern may act as a "dynamic template", providing local cues which trigger behavioural acts appropriate to colony needs, which in turn may feed back to changes in the gas template. In particular, wall building occurs along lines of similar concentration for a variety of nest geometries; there is surprising convergence between the period of cycles of synchronously active ants and the time taken for CO2 levels to equilibrate; and the qualitatively distinct regions of the "dynamic template" correspond to regions occupied by different groups of ants.

  13. Buteo Nesting Ecology: Evaluating Nesting of Swainson’s Hawks in the Northern Great Plains

    PubMed Central

    Inselman, Will M.; Datta, Shubham; Jenks, Jonathan A.; Jensen, Kent C.; Grovenburg, Troy W.

    2015-01-01

    Swainson’s hawks (Buteo swainsoni) are long-distance migratory raptors that nest primarily in isolated trees located in areas of high grassland density. In recent years, anthropogenic conversion of grassland habitat has raised concerns about the status of the breeding population in the northern Great Plains. In 2013, we initiated a study to investigate the influence of extrinsic factors influencing Swainson’s hawk nesting ecology in north-central South Dakota and south-central North Dakota. Using ground and aerial surveys, we located and monitored nesting Swainson’s hawk pairs: 73 in 2013 and 120 in 2014. We documented 98 successful breeding attempts that fledged 163 chicks; 1.52 and 1.72 fledglings per successful nest in 2013 and 2014, respectively. We used Program MARK to evaluate the influence of land cover on nest survival. The top model, S Dist2Farm+%Hay, indicated that nest survival (fledging at least one chick) decreased as nests were located farther from farm sites and as the percent of hay cover increased within 1200-m of the nest site (34.4%; 95% CI = 27.6%–42.3%). We used logistic regression analysis to evaluate the influence of landscape variables on nest-site selection; Swainson’s hawks selected for nest sites located closer to roads. We suggest that tree belts associated with farm sites, whether occupied or not, provide critical breeding sites for Swainson’s hawks. Additionally, poor breeding success may be related to the late migratory behavior of this species which requires them to occupy marginal habitat due to other raptors occupying the most suitable habitat prior to Swainson’s hawks arriving to the breeding grounds. PMID:26327440

  14. Determinants of mallard and gadwall nesting on constructed islands in North Dakota

    USGS Publications Warehouse

    Shaffer, Terry L.; Dahl, Ann L.; Reynolds, Ronald E.; Baer, Kathy L.; Johnson, Michael A.; Sargeant, Glen A.

    2006-01-01

    Constructed islands with adequate nesting cover provide secure nesting sites for ducks because islands restrict access by mammalian predators. These islands are costly to construct and should be placed in areas that ensure the greatest use by nesting ducks. We studied mallard (Anas platyrhynchos) and gadwall (A. strepera) nesting on constructed islands in North Dakota in 1996 (n = 20) and 1997 (n = 22) to evaluate factors—particularly amount of perennial grass cover in the surrounding landscape and density of breeding pairs—that possibly influence numbers of initiated nests. We also examined effects of island characteristics, such as island vegetation, on numbers of nests. Numbers of mallard and gadwall nests on islands were negatively related to amounts of perennial grass cover in the surrounding uplands. Numbers of mallard nests were positively related to percentages of tall dense cover on islands. We found no effects of breeding-pair density on numbers of nests initiated by either species, possibly because breeding pairs were abundant on all study sites. Percent shrub cover on islands was a better predictor of island use than was percent tall dense cover. Island use by these species increased with island age and distance from mainland shore. Amounts of perennial cover in landscapes should be primary considerations in determining where to build islands. Our data suggest that use of islands by nesting mallards and gadwalls is greatest in landscapes with little perennial grass cover (i.e., high amounts of cropland). Other researchers documented a positive relation between nest success in upland covers and amount of perennial grass cover in the landscape. Therefore, islands constructed in landscapes with little perennial cover should provide greater gains in duck recruitment rates than islands constructed in landscapes with greater amounts of perennial grass cover.

  15. Probability of detection of nests and implications for survey design

    USGS Publications Warehouse

    Smith, P.A.; Bart, J.; Lanctot, Richard B.; McCaffery, B.J.; Brown, S.

    2009-01-01

    Surveys based on double sampling include a correction for the probability of detection by assuming complete enumeration of birds in an intensively surveyed subsample of plots. To evaluate this assumption, we calculated the probability of detecting active shorebird nests by using information from observers who searched the same plots independently. Our results demonstrate that this probability varies substantially by species and stage of the nesting cycle but less by site or density of nests. Among the species we studied, the estimated single-visit probability of nest detection during the incubation period varied from 0.21 for the White-rumped Sandpiper (Calidris fuscicollis), the most difficult species to detect, to 0.64 for the Western Sandpiper (Calidris mauri), the most easily detected species, with a mean across species of 0.46. We used these detection probabilities to predict the fraction of persistent nests found over repeated nest searches. For a species with the mean value for detectability, the detection rate exceeded 0.85 after four visits. This level of nest detection was exceeded in only three visits for the Western Sandpiper, but six to nine visits were required for the White-rumped Sandpiper, depending on the type of survey employed. Our results suggest that the double-sampling method's requirement of nearly complete counts of birds in the intensively surveyed plots is likely to be met for birds with nests that survive over several visits of nest searching. Individuals with nests that fail quickly or individuals that do not breed can be detected with high probability only if territorial behavior is used to identify likely nesting pairs. ?? The Cooper Ornithological Society, 2009.

  16. Catastrophic health expenditure: a comparative analysis of empty-nest and non-empty-nest households with seniors in Shandong, China.

    PubMed

    Yang, Tingting; Chu, Jie; Zhou, Chengchao; Medina, Alexis; Li, Cuicui; Jiang, Shan; Zheng, Wengui; Sun, Liyuan; Liu, Jing

    2016-07-05

    The aim of this study was to compare the catastrophic health expenditure (CHE) prevalence and its determinants between empty-nest and non-empty-nest elderly households. Shandong province of China. A total of 2761 elderly households are included in the analysis. CHE incidence among elderly households was 44.9%. The CHE incidence of empty-nest singles (59.3%, p=0.000, OR=3.19) and empty-nest couples (52.9%, p=0.000, OR=2.45) are both statistically higher than that of non-empty-nest elderly households (31.4%). An inverse association was observed between CHE incidence and income level in all elderly household types. Factors including 1 or more household elderly members with non-communicable chronic diseases in the past 6 months, 1 or more elderly household members being hospitalised in the past year and lower household income, are significant risk factors for CHE in all 3 household types (p<0.05). Health insurance status was found to be a significant determinant of CHE among empty-nest singles and non-empty-nest households (p<0.05). CHE incidence among elderly households is high in China. Empty-nest households are at higher risk for CHE than non-empty-nest households. Based on these findings, we suggest that special insurance be developed to broaden the coverage of health services and heighten the reimbursement rate for empty-nest elderly in the existing health insurance schemes. Financial and social protection interventions are also essential for identified at-risk subgroups among different types of elderly households. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  17. Catastrophic health expenditure: a comparative analysis of empty-nest and non-empty-nest households with seniors in Shandong, China

    PubMed Central

    Yang, Tingting; Chu, Jie; Zhou, Chengchao; Medina, Alexis; Li, Cuicui; Jiang, Shan; Zheng, Wengui; Sun, Liyuan; Liu, Jing

    2016-01-01

    Objective The aim of this study was to compare the catastrophic health expenditure (CHE) prevalence and its determinants between empty-nest and non-empty-nest elderly households. Setting Shandong province of China. Participants A total of 2761 elderly households are included in the analysis. Results CHE incidence among elderly households was 44.9%. The CHE incidence of empty-nest singles (59.3%, p=0.000, OR=3.19) and empty-nest couples (52.9%, p=0.000, OR=2.45) are both statistically higher than that of non-empty-nest elderly households (31.4%). An inverse association was observed between CHE incidence and income level in all elderly household types. Factors including 1 or more household elderly members with non-communicable chronic diseases in the past 6 months, 1 or more elderly household members being hospitalised in the past year and lower household income, are significant risk factors for CHE in all 3 household types (p<0.05). Health insurance status was found to be a significant determinant of CHE among empty-nest singles and non-empty-nest households (p<0.05). Conclusions CHE incidence among elderly households is high in China. Empty-nest households are at higher risk for CHE than non-empty-nest households. Based on these findings, we suggest that special insurance be developed to broaden the coverage of health services and heighten the reimbursement rate for empty-nest elderly in the existing health insurance schemes. Financial and social protection interventions are also essential for identified at-risk subgroups among different types of elderly households. PMID:27381206

  18. The common redstart as a suitable model to study cuckoo-host coevolution in a unique ecological context.

    PubMed

    Samaš, Peter; Rutila, Jarkko; Grim, Tomáš

    2016-11-25

    Co-evolutionary arms-races result in spatio-temporally dynamic relationships between interacting species, e.g., brood parasites and their avian hosts. However, majority of avian co-evolutionary studies are limited to "snap-shots" of a single breeding season in an open-nesting host. In a long-term study (11 breeding seasons), we explored a unique system between the brood parasitic common cuckoo (Cuculus canorus) and its host, the common redstart (Phoenicurus phoenicurus) which is exceptional among all cuckoo hosts due to being a cavity nester. Conditions in cavities are different from open nests, e.g., lower risks of predation, more favourable microclimate, increased risks of unsuccessful eviction of host offspring by the cuckoo nestling. Different conditions in cavities thus can be expected to shape parasite-host coevolution differently from what is typically studied in open nesting hosts. In our highly parasitised nest-box population (32.5%, n = 569 nests) only 35.7% of cuckoo eggs were laid into the nest cup and incubated by redstarts. Host nests shifted availability to later into the breeding season from 2006 to 2016 and cuckoos followed this trend by also shifting their timing of parasitism. Although previous studies revealed that redstarts selectively eject experimental non-mimetic eggs (desertion was not a specific response to foreign eggs), the hosts never ejected naturally-laid cuckoo eggs or cuckoo eggs cross-fostered into naturally non-parasitised nests. We solve the long-standing debate about the origin of cuckoo eggs found on the nest rim: we gained the first direct video-recording evidence that eggs found on the nest rim were mislaid by parasites and not ejected by hosts. Naturally-parasitised nests were deserted more often (18.6%) than control non-parasitized nests (5.6%) or nests artificially parasitised by us (1.4%). This suggests that the sight of the laying cuckoo female is the primary cue that triggers egg rejection (by desertion) in this host. Review of data from this and other study sites (10 populations, n = 853 experiments) demonstrates high variability in rejection rates and shows that populations facing higher parasitism rates reject parasitic eggs with higher frequencies. Surprisingly, cuckoo chicks either growing solitarily or with redstart chicks did not differ in their fledging success. We suggest that the redstart is an ideal model system to study the flexibility and limits of brood parasite-host co-evolution in an extreme ecological setting.

  19. Birds' nesting parameters in four forest types in the Pantanal wetland.

    PubMed

    Pinho, J B; Marini, M A

    2014-11-01

    We tested the heterogeneity/productivity hypothesis with respect to the abundance and richness of birds and the vegetation density hypothesis with respect to birds' nest predation rates, and determined the relative importance of forested vegetation formations for the conservation of birds in the Pantanal. We estimated the apparent nesting success, and the abundance and richness of nesting birds' in four forest types, by monitoring nests during two reproductive seasons in four forested physiognomies (two high productivity/heterogeneity evergreen forests = Cambará and Landi; two low productivity/heterogeneity dry forests = Cordilheira and Carvoeiro) in the Pantanal wetland in Poconé, State of Mato Grosso, Brazil. We found 381 nests of 46 species (35 Passeriformes and 11 non-Passeriformes) in the four forest types. Of these, we monitored 220 active nests belonging to 44 species, 101 during the reproductive season of 2001 and 119 in 2002. We supported the productivity/heterogeneity hypothesis since the two evergreen forests had higher nest abundance and one of them (Cambará) had higher nesting species richness than the dry forests. The number of nests found in each habitat differed with most nests monitored in the Cambará forest (82%), followed by Landi (9%), Cordilheira (6%) and Carvoeiro (3%) forests. The total number of nests monitored was significantly higher in evergreen forests than in dry forests. Also, more species nested in evergreen (37 species) than in dry (16 species) forests. A Correspondence Analysis revealed that only Carvoeiros had a different nesting bird community. The overall apparent nesting success of 220 nests was 26.8%. We did not support the vegetation density hypothesis since nest predation rates were similar between evergreen (73.5%) and dry (70%) forests, and were higher in the Landi (85%) than in the other three forests (69.2 to 72.2%). Our data indicate that Cambará forests seem to be a key nesting habitat for many bird species of the Pantanal. If this local pattern also occurs in other regions of the Pantanal, the use and management of Cambará forests might prove to be important for the conservation of forest birds of this biome. However, conflicting results with other taxonomic groups show that conservation measures for these forests and land use policies should be based on a more complete biodiversity evaluation of the region.

  20. Breeding biology and success of a reintroduced population of the critically endangered Puaiohi (Myadestes palmeri)

    USGS Publications Warehouse

    Tweed, E.J.; Foster, J.T.; Woodworth, B.L.; Monahan, W.B.; Kellerman, J.L.; Lieberman, A.

    2006-01-01

    The ultimate success of reintroduction programs for endangered species depends on the ability of reintroduced animals to breed in the wild. We studied the nesting success and breeding biology of a reintroduced population of Puaiohi (Myadestes palmeri) on the island of Kaua'i, Hawaii. Thirty-four captive-bred Puaiohi were released into the Alaka'i Swamp in 1999-2001 and monitored using radiotelemetry. Ten females and two males paired with wild and other released birds, including one polygynous trio. From March to September, 31 nests were built. Mean clutch size was 2.0 eggs, daily nest survival was 0.97 ± 0.01 (mean ± SE) and overall nest success was 0.40 ± 0.02. We confirmed predation, most probably by rats (Rattus spp.), as the greatest cause of nest failure, occurring at 38% of active nests with known fates, and causing the death of two nesting adult females. Ground-based rodent control proved ineffective at protecting nest attempts. Successful nests fledged an average of 1.4 young each (n = 10), and 85% of fledglings survived at least two weeks. Importantly, breeding behavior and success were comparable to those of wild Puaiohi. This is the first record of breeding in the wild from captive-bred endangered Hawaiian passerines. The ability of captive-bred Puaiohi to survive and breed successfully in the wild bodes well for future releases of this and other endangered passerines, but high predation rates on nests and nesting females highlights the importance of maintaining and restoring safe habitat for recovery.

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