Sample records for animals sampled based

  1. Relative sensitivity of conventional and real-time PCR assays for detection of SFG Rickettsia in blood and tissue samples from laboratory animals.

    PubMed

    Zemtsova, Galina E; Montgomery, Merrill; Levin, Michael L

    2015-01-01

    Studies on the natural transmission cycles of zoonotic pathogens and the reservoir competence of vertebrate hosts require methods for reliable diagnosis of infection in wild and laboratory animals. Several PCR-based applications have been developed for detection of infections caused by Spotted Fever group Rickettsia spp. in a variety of animal tissues. These assays are being widely used by researchers, but they differ in their sensitivity and reliability. We compared the sensitivity of five previously published conventional PCR assays and one SYBR green-based real-time PCR assay for the detection of rickettsial DNA in blood and tissue samples from Rickettsia- infected laboratory animals (n = 87). The real-time PCR, which detected rickettsial DNA in 37.9% of samples, was the most sensitive. The next best were the semi-nested ompA assay and rpoB conventional PCR, which detected as positive 18.4% and 14.9% samples respectively. Conventional assays targeting ompB, gltA and hrtA genes have been the least sensitive. Therefore, we recommend the SYBR green-based real-time PCR as a tool for the detection of rickettsial DNA in animal samples due to its higher sensitivity when compared to more traditional assays.

  2. Relative Sensitivity of Conventional and Real-Time PCR Assays for Detection of SFG Rickettsia in Blood and Tissue Samples from Laboratory Animals

    PubMed Central

    Zemtsova, Galina E.; Montgomery, Merrill; Levin, Michael L.

    2015-01-01

    Studies on the natural transmission cycles of zoonotic pathogens and the reservoir competence of vertebrate hosts require methods for reliable diagnosis of infection in wild and laboratory animals. Several PCR-based applications have been developed for detection of infections caused by Spotted Fever group Rickettsia spp. in a variety of animal tissues. These assays are being widely used by researchers, but they differ in their sensitivity and reliability. We compared the sensitivity of five previously published conventional PCR assays and one SYBR green-based real-time PCR assay for the detection of rickettsial DNA in blood and tissue samples from Rickettsia- infected laboratory animals (n = 87). The real-time PCR, which detected rickettsial DNA in 37.9% of samples, was the most sensitive. The next best were the semi-nested ompA assay and rpoB conventional PCR, which detected as positive 18.4% and 14.9% samples respectively. Conventional assays targeting ompB, gltA and hrtA genes have been the least sensitive. Therefore, we recommend the SYBR green-based real-time PCR as a tool for the detection of rickettsial DNA in animal samples due to its higher sensitivity when compared to more traditional assays. PMID:25607846

  3. A parametric generalization of the Hayne estimator for line transect sampling

    USGS Publications Warehouse

    Burnham, Kenneth P.

    1979-01-01

    The Hayne model for line transect sampling is generalized by using an elliptical (rather than circular) flushing model for animal detection. By assuming the ration of major and minor axes lengths is constant for all animals, a model results which allows estimation of population density based directly upon sighting distances and sighting angles. The derived estimator of animal density is a generalization of the Hayne estimator for line transect sampling.

  4. Salmonella and antimicrobial resistance in an animal-based agriculture river system.

    PubMed

    Palhares, Julio Cesar Pascale; Kich, Jalusa D; Bessa, Marjo C; Biesus, Luiza L; Berno, Lais G; Triques, Nelise J

    2014-02-15

    The aim of this study was to examine the Salmonella serovars and antimicrobial resistance within an animal-based agriculture river system. The study area consisted of a 1,345 ha upper part of Pinhal catchment. A total of 384 samples were collected in four years of monitoring. Salmonella was isolated from 241 samples (62.7%), resulting in 324 isolates. The highest number of Salmonella sp. occurred in samples associated with sites with high stoking density animal unit per hectare. It was possible to demonstrate the variability of serovars in the study area: 30 different serovars were found and at least 11 per monitoring site. Thirty-three potentially related isolates were genotyped by PFGE, one major clone was observed in serovar Typhimurium, which occurred in animal feces (swine and bovine), and different sites and samplings proving the cross-contamination and persistence of this specific clone. Among 180 isolates submitted to an antimicrobial susceptibility test, 50.5% were susceptible to all 21 antimicrobials tested and 54 different profiles were found. In the current study, 49.5% of the tested isolates were resistant to at least one antimicrobial, and multi-resistance occurred in 18% of isolates. Results indicate a close interaction between animal-based agriculture, Salmonella, and antimicrobial resistance. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Childhood animal cruelty methods and their link to adult interpersonal violence.

    PubMed

    Henderson, Brandy B; Hensley, Christopher; Tallichet, Suzanne E

    2011-07-01

    Recent research has begun to establish a relationship between childhood acts of animal cruelty and later violence against humans. However, few studies have focused on the influence of animal cruelty methods on later interpersonal violence. In a replication of a study by Hensley and Tallichet (2009) and based on a sample of 180 inmates at medium- and maximum-security prisons in a Southern state, the present study examines the relationship between several retrospectively identified animal cruelty methods (drowned, hit, shot, kicked, choked, burned, and sex) and interpersonal violence committed against humans. Four out of 5 inmates reported hitting animals. Over one third of the sample chose to shoot or kick animals, while 1 in 5 had sex with them. Less then one fifth of the sample drowned or choked animals, while less than one sixth of the inmates burned animals. Regression analyses revealed that the age at which offenders began committing animal cruelty and having sex with animals were predictive of adult interpersonal violence.

  6. Active animal health surveillance in European Union Member States: gaps and opportunities.

    PubMed

    Bisdorff, B; Schauer, B; Taylor, N; Rodríguez-Prieto, V; Comin, A; Brouwer, A; Dórea, F; Drewe, J; Hoinville, L; Lindberg, A; Martinez Avilés, M; Martínez-López, B; Peyre, M; Pinto Ferreira, J; Rushton, J; VAN Schaik, G; Stärk, K D C; Staubach, C; Vicente-Rubiano, M; Witteveen, G; Pfeiffer, D; Häsler, B

    2017-03-01

    Animal health surveillance enables the detection and control of animal diseases including zoonoses. Under the EU-FP7 project RISKSUR, a survey was conducted in 11 EU Member States and Switzerland to describe active surveillance components in 2011 managed by the public or private sector and identify gaps and opportunities. Information was collected about hazard, target population, geographical focus, legal obligation, management, surveillance design, risk-based sampling, and multi-hazard surveillance. Two countries were excluded due to incompleteness of data. Most of the 664 components targeted cattle (26·7%), pigs (17·5%) or poultry (16·0%). The most common surveillance objectives were demonstrating freedom from disease (43·8%) and case detection (26·8%). Over half of components applied risk-based sampling (57·1%), but mainly focused on a single population stratum (targeted risk-based) rather than differentiating between risk levels of different strata (stratified risk-based). About a third of components were multi-hazard (37·3%). Both risk-based sampling and multi-hazard surveillance were used more frequently in privately funded components. The study identified several gaps (e.g. lack of systematic documentation, inconsistent application of terminology) and opportunities (e.g. stratified risk-based sampling). The greater flexibility provided by the new EU Animal Health Law means that systematic evaluation of surveillance alternatives will be required to optimize cost-effectiveness.

  7. A TaqMan real-time PCR method based on alternative oxidase genes for detection of plant species in animal feed samples.

    PubMed

    Campos, Maria Doroteia; Valadas, Vera; Campos, Catarina; Morello, Laura; Braglia, Luca; Breviario, Diego; Cardoso, Hélia G

    2018-01-01

    Traceability of processed food and feed products has been gaining importance due to the impact that those products can have on human/animal health and to the associated economic and legal concerns, often related to adulterations and frauds as it can be the case for meat and milk. Despite mandatory traceability requirements for the analysis of feed composition, few reliable and accurate methods are presently available to enforce the legislative frame and allow the authentication of animal feeds. In this study, nine sensitive and species-specific real-time PCR TaqMan MGB assays are described for plant species detection in animal feed samples. The method is based on selective real-time qPCR (RT-qPCR) amplification of target genes belonging to the alternative oxidase (AOX) gene family. The plant species selected for detection in feed samples were wheat, maize, barley, soybean, rice and sunflower as common components of feeds, and cotton, flax and peanut as possible undesirable contaminants. The obtained results were compared with end-point PCR methodology. The applicability of the AOX TaqMan assays was evaluated through the screening of commercial feed samples, and by the analysis of plant mixtures with known composition. The RT-qPCR methodology allowed the detection of the most abundant species in feeds but also the identification of contaminant species present in lower amounts, down to 1% w/w. AOX-based methodology provides a suitable molecular marker approach to ascertain plant species composition of animal feed samples, thus supporting feed control and enforcement of the feed sector and animal production.

  8. Colorado animal-based plague surveillance systems: relationships between targeted animal species and prediction efficacy of areas at risk for humans.

    PubMed

    Lowell, Jennifer L; Eisen, Rebecca J; Schotthoefer, Anna M; Xiaocheng, Liang; Montenieri, John A; Tanda, Dale; Pape, John; Schriefer, Martin E; Antolin, Michael F; Gage, Kenneth L

    2009-06-01

    Human plague risks (Yersinia pestis infection) are greatest when epizootics cause high mortality among this bacterium's natural rodent hosts. Therefore, health departments in plague-endemic areas commonly establish animal-based surveillance programs to monitor Y. pestis infection among plague hosts and vectors. The primary objectives of our study were to determine whether passive animal-based plague surveillance samples collected in Colorado from 1991 to 2005 were sampled from high human plague risk areas and whether these samples provided information useful for predicting human plague case locations. By comparing locations of plague-positive animal samples with a previously constructed GIS-based plague risk model, we determined that the majority of plague-positive Gunnison's prairie dogs (100%) and non-prairie dog sciurids (85.82%), and moderately high percentages of sigmodontine rodents (71.4%), domestic cats (69.3%), coyotes (62.9%), and domestic dogs (62.5%) were recovered within 1 km of the nearest area posing high peridomestic risk to humans. In contrast, the majority of white-tailed prairie dog (66.7%), leporid (cottontailed and jack rabbits) (71.4%), and black-tailed prairie dog (93.0%) samples originated more than 1 km from the nearest human risk habitat. Plague-positive animals or their fleas were rarely (one of 19 cases) collected within 2 km of a case exposure site during the 24 months preceding the dates of illness onset for these cases. Low spatial accuracy for identifying epizootic activity prior to human plague cases suggested that other mammalian species or their fleas are likely more important sources of human infection in high plague risk areas. To address this issue, epidemiological observations and multi-locus variable number tandem repeat analyses (MLVA) were used to preliminarily identify chipmunks as an under-sampled, but potentially important, species for human plague risk in Colorado.

  9. Hierarchical models of animal abundance and occurrence

    USGS Publications Warehouse

    Royle, J. Andrew; Dorazio, R.M.

    2006-01-01

    Much of animal ecology is devoted to studies of abundance and occurrence of species, based on surveys of spatially referenced sample units. These surveys frequently yield sparse counts that are contaminated by imperfect detection, making direct inference about abundance or occurrence based on observational data infeasible. This article describes a flexible hierarchical modeling framework for estimation and inference about animal abundance and occurrence from survey data that are subject to imperfect detection. Within this framework, we specify models of abundance and detectability of animals at the level of the local populations defined by the sample units. Information at the level of the local population is aggregated by specifying models that describe variation in abundance and detection among sites. We describe likelihood-based and Bayesian methods for estimation and inference under the resulting hierarchical model. We provide two examples of the application of hierarchical models to animal survey data, the first based on removal counts of stream fish and the second based on avian quadrat counts. For both examples, we provide a Bayesian analysis of the models using the software WinBUGS.

  10. Reproducibility of preclinical animal research improves with heterogeneity of study samples

    PubMed Central

    Vogt, Lucile; Sena, Emily S.; Würbel, Hanno

    2018-01-01

    Single-laboratory studies conducted under highly standardized conditions are the gold standard in preclinical animal research. Using simulations based on 440 preclinical studies across 13 different interventions in animal models of stroke, myocardial infarction, and breast cancer, we compared the accuracy of effect size estimates between single-laboratory and multi-laboratory study designs. Single-laboratory studies generally failed to predict effect size accurately, and larger sample sizes rendered effect size estimates even less accurate. By contrast, multi-laboratory designs including as few as 2 to 4 laboratories increased coverage probability by up to 42 percentage points without a need for larger sample sizes. These findings demonstrate that within-study standardization is a major cause of poor reproducibility. More representative study samples are required to improve the external validity and reproducibility of preclinical animal research and to prevent wasting animals and resources for inconclusive research. PMID:29470495

  11. Automated DBS microsampling, microscale automation and microflow LC-MS for therapeutic protein PK.

    PubMed

    Zhang, Qian; Tomazela, Daniela; Vasicek, Lisa A; Spellman, Daniel S; Beaumont, Maribel; Shyong, BaoJen; Kenny, Jacqueline; Fauty, Scott; Fillgrove, Kerry; Harrelson, Jane; Bateman, Kevin P

    2016-04-01

    Reduce animal usage for discovery-stage PK studies for biologics programs using microsampling-based approaches and microscale LC-MS. We report the development of an automated DBS-based serial microsampling approach for studying the PK of therapeutic proteins in mice. Automated sample preparation and microflow LC-MS were used to enable assay miniaturization and improve overall assay throughput. Serial sampling of mice was possible over the full 21-day study period with the first six time points over 24 h being collected using automated DBS sample collection. Overall, this approach demonstrated comparable data to a previous study using single mice per time point liquid samples while reducing animal and compound requirements by 14-fold. Reduction in animals and drug material is enabled by the use of automated serial DBS microsampling for mice studies in discovery-stage studies of protein therapeutics.

  12. A Story-Based Simulation for Teaching Sampling Distributions

    ERIC Educational Resources Information Center

    Turner, Stephen; Dabney, Alan R.

    2015-01-01

    Statistical inference relies heavily on the concept of sampling distributions. However, sampling distributions are difficult to teach. We present a series of short animations that are story-based, with associated assessments. We hope that our contribution can be useful as a tool to teach sampling distributions in the introductory statistics…

  13. Machine vision application in animal trajectory tracking.

    PubMed

    Koniar, Dušan; Hargaš, Libor; Loncová, Zuzana; Duchoň, František; Beňo, Peter

    2016-04-01

    This article was motivated by the doctors' demand to make a technical support in pathologies of gastrointestinal tract research [10], which would be based on machine vision tools. Proposed solution should be less expensive alternative to already existing RF (radio frequency) methods. The objective of whole experiment was to evaluate the amount of animal motion dependent on degree of pathology (gastric ulcer). In the theoretical part of the article, several methods of animal trajectory tracking are presented: two differential methods based on background subtraction, the thresholding methods based on global and local threshold and the last method used for animal tracking was the color matching with a chosen template containing a searched spectrum of colors. The methods were tested offline on five video samples. Each sample contained situation with moving guinea pig locked in a cage under various lighting conditions. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  14. Comparative diagnostic efficacy of recombinant LLO and PI-PLC-based ELISAs for detection of listeriosis in animals.

    PubMed

    Suryawanshi, Rahul D; Malik, Satya Veer Singh; Jayarao, Bhushan; Chaudhari, Sandeep P; Savage, Emily; Vergis, Jess; Kurkure, Nitin V; Barbuddhe, Sukhadeo B; Rawool, Deepak B

    2017-06-01

    The present study for the first time evaluates the serodiagnostic efficacy of two recombinant antigens namely, listeriolysin O (rLLO) and phosphatidyl-inositol phospholipase C (rPI-PLC). Indirect ELISA with the above recombinant antigens was used on samples collected from bovines (n=106), goats (n=138) and pigs (n=92) having either a history of abortion, emaciation and/or apparently healthy animals. Isolation of Listeria was attempted from the blood samples using USDA-FSIS method. On screening of test sera by rLLO-based ELISA, antibodies against anti-listeriolysin O (ALLO) were observed in goats (22.46%), bovines (15.10%) and pigs (16.31%). As advocated, after adsorption of positive serum samples with streptolysin O (SLO), the seropositivity for ALLO was marginally reduced (p>0.05) in goats (21.73%) and bovines (10.38%), whereas, in pigs the reduction (5.43%) was significant (p<0.05). On the contrary, rPI-PLC-based ELISA revealed higher non-specific seropositivity for antilisterial antibodies in goats (45.65%), bovines (31.13%) and pigs (8.69%). Further, on comparing the seropositivity with isolation rate, of the 16 animals that were culturally-positive for L. monocytogenes, 15 showed ALLO positivity in unadsorbed as well as SLO-adsorbed sera by rLLO-based ELISA, however, rPI-PLC-based ELISA could detect seropositivity in only 5 animals. Moreover, rPI-PLC-based ELISA also showed seropositivity in those animals (7/30) that were culturally positive for other Listeria spp. In conclusion, rLLO can serve as a better antigen than rPI-PLC in ELISA for the serodiagnosis of listeriosis in animals; however, prior adsorption of test sera with SLO is required to avoid false positive results. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Genie Inference Engine Rule Writer’s Guide.

    DTIC Science & Technology

    1987-08-01

    33 APPENDIX D. Animal Bootstrap File.............................................................. 39...APPENDIX E. Sample Run of Animal Identification Expert System.......................... 43 APPENDIX F. Numeric Test Knowledge Base...and other data s.tructures stored in the knowledge base (KB), queries the user for input, and draws conclusions. Genie (GENeric Inference Engine) is

  16. Employment Opportunities in Applied Biological and Agricultural Occupations in the Metropolitan Area of Chicago.

    ERIC Educational Resources Information Center

    Thomas, Hollie B.; Neavill, Arthur

    Based on questionnaire data collected from a sample of employers, this phase of a larger research project ascertained employment opportunities in the area of applied biological and agricultural occupations in the metropolitan area of Chicago. Specific fields of business surveyed by stratified random sample were animal care, animal health care,…

  17. Novel rope-based sampling of classical swine fever shedding in a group of wild boar showing low contagiosity upon experimental infection with a classical swine fever field strain of genotype 2.3.

    PubMed

    Mouchantat, Susan; Globig, Anja; Böhle, Wolfgang; Petrov, Anja; Strebelow, Heinz-Günther; Mettenleiter, Thomas C; Depner, Klaus

    2014-06-04

    Several classical swine fever (CSF) epidemics in wild boar and domestic pigs in Europe during the last decades have been caused by CSF virus (CSFV) strains of genotype 2.3. This genotype is known to be virulent leading to high morbidity and mortality. We experimentally infected two eight months old wild boar with 10(5,5) TCID50 of CSFV genotype 2.3 and kept the animals together with five noninoculated wild boar of the same age. Our original purpose was to evaluate a non-invasive sampling method based on saliva collection using "rope-in-a-bait" sampling baits. While expecting high morbidity, high level of virus shedding and some mortality, we actually observed a subclinical course of infection with an unexpected low contagiosity. The two inoculated animals infected only three contact animals while two contact animals remained uninfected. These findings substantially add to our epidemiological understanding of CSFV circulation in wild boar populations. CSFV infected animals older than six months and in good condition may not shed sufficient virus to transmit infection to all seronegative in-contact animals. The contagiosity in relation to the animal's age is discussed. This supports the hypothesis of silent perpetuation of CSFV in wild boar populations for several months if the wild boar density is sufficiently high. The feasibility of the "rope-in-a-bait" sampling method could be proven during the short viraemic phase of infected animals during the second week of infection. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Sample preparation combined with electroanalysis to improve simultaneous determination of antibiotics in animal derived food samples.

    PubMed

    da Silva, Wesley Pereira; de Oliveira, Luiz Henrique; Santos, André Luiz Dos; Ferreira, Valdir Souza; Trindade, Magno Aparecido Gonçalves

    2018-06-01

    A procedure based on liquid-liquid extraction (LLE) and phase separation using magnetically stirred salt-induced high-temperature liquid-liquid extraction (PS-MSSI-HT-LLE) was developed to extract and pre-concentrate ciprofloxacin (CIPRO) and enrofloxacin (ENRO) from animal food samples before electroanalysis. Firstly, simple LLE was used to extract the fluoroquinolones (FQs) from animal food samples, in which dilution was performed to reduce interference effects to below a tolerable threshold. Then, adapted PS-MSSI-HT-LLE protocols allowed re-extraction and further pre-concentration of target analytes in the diluted acid samples for simultaneous electrochemical quantification at low concentration levels. To improve the peak separation, in simultaneous detection, a baseline-corrected second-order derivative approach was processed. These approaches allowed quantification of target FQs from animal food samples spiked at levels of 0.80 to 2.00 µmol L -1 in chicken meat, with recovery values always higher than 80.5%, as well as in milk samples spiked at 4.00 µmol L -1 , with recovery values close to 70.0%. Copyright © 2018 Elsevier Ltd. All rights reserved.

  19. Policy-driven development of cost-effective, risk-based surveillance strategies.

    PubMed

    Reist, M; Jemmi, T; Stärk, K D C

    2012-07-01

    Animal health and residue surveillance verifies the good health status of the animal population, thereby supporting international free trade of animals and animal products. However, active surveillance is costly and time-consuming. The development of cost-effective tools for animal health and food hazard surveillance is therefore a priority for decision-makers in the field of veterinary public health. The assumption of this paper is that outcome-based formulation of standards, legislation leaving room for risk-based approaches and close collaboration and a mutual understanding and exchange between scientists and policy makers are essential for cost-effective surveillance. We illustrate this using the following examples: (i) a risk-based sample size calculation for surveys to substantiate freedom from diseases/infection, (ii) a cost-effective national surveillance system for Bluetongue using scenario tree modelling and (iii) a framework for risk-based residue monitoring. Surveys to substantiate freedom from infectious bovine rhinotracheitis and enzootic bovine leucosis between 2002 and 2009 saved over 6 million € by applying a risk-based sample size calculation approach, and by taking into account prior information from repeated surveys. An open, progressive policy making process stimulates research and science to develop risk-based and cost-efficient survey methodologies. Early involvement of policy makers in scientific developments facilitates implementation of new findings and full exploitation of benefits for producers and consumers. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Efficient Stochastic Rendering of Static and Animated Volumes Using Visibility Sweeps.

    PubMed

    von Radziewsky, Philipp; Kroes, Thomas; Eisemann, Martin; Eisemann, Elmar

    2017-09-01

    Stochastically solving the rendering integral (particularly visibility) is the de-facto standard for physically-based light transport but it is computationally expensive, especially when displaying heterogeneous volumetric data. In this work, we present efficient techniques to speed-up the rendering process via a novel visibility-estimation method in concert with an unbiased importance sampling (involving environmental lighting and visibility inside the volume), filtering, and update techniques for both static and animated scenes. Our major contributions include a progressive estimate of partial occlusions based on a fast sweeping-plane algorithm. These occlusions are stored in an octahedral representation, which can be conveniently transformed into a quadtree-based hierarchy suited for a joint importance sampling. Further, we propose sweep-space filtering, which suppresses the occurrence of fireflies and investigate different update schemes for animated scenes. Our technique is unbiased, requires little precomputation, is highly parallelizable, and is applicable to a various volume data sets, dynamic transfer functions, animated volumes and changing environmental lighting.

  1. Sampling considerations for disease surveillance in wildlife populations

    USGS Publications Warehouse

    Nusser, S.M.; Clark, W.R.; Otis, D.L.; Huang, L.

    2008-01-01

    Disease surveillance in wildlife populations involves detecting the presence of a disease, characterizing its prevalence and spread, and subsequent monitoring. A probability sample of animals selected from the population and corresponding estimators of disease prevalence and detection provide estimates with quantifiable statistical properties, but this approach is rarely used. Although wildlife scientists often assume probability sampling and random disease distributions to calculate sample sizes, convenience samples (i.e., samples of readily available animals) are typically used, and disease distributions are rarely random. We demonstrate how landscape-based simulation can be used to explore properties of estimators from convenience samples in relation to probability samples. We used simulation methods to model what is known about the habitat preferences of the wildlife population, the disease distribution, and the potential biases of the convenience-sample approach. Using chronic wasting disease in free-ranging deer (Odocoileus virginianus) as a simple illustration, we show that using probability sample designs with appropriate estimators provides unbiased surveillance parameter estimates but that the selection bias and coverage errors associated with convenience samples can lead to biased and misleading results. We also suggest practical alternatives to convenience samples that mix probability and convenience sampling. For example, a sample of land areas can be selected using a probability design that oversamples areas with larger animal populations, followed by harvesting of individual animals within sampled areas using a convenience sampling method.

  2. First Results in the Use of Bovine Ear Notch Tag for Bovine Viral Diarrhoea Virus Detection and Genetic Analysis

    PubMed Central

    Quinet, Christian; Czaplicki, Guy; Dion, Elise; Dal Pozzo, Fabiana; Kurz, Anke; Saegerman, Claude

    2016-01-01

    Background Infection due to bovine viral diarrhoea virus (BVDV) is endemic in most cattle-producing countries throughout the world. The key elements of a BVDV control programme are biosecurity, elimination of persistently infected animals and surveillance. Bovine viral diarrhoea (BVD) is a notifiable disease in Belgium and an official eradication programme started from January 2015, based on testing ear notches sampled during the official identification and registration of calves at birth. An antigen-capture ELISA test based on the detection of BVDV Erns protein is used. Ear notch sample may also be used to characterize the genotype of the calf when appropriate elution/dilution buffer is added. Both BVDV antigen-ELISA analysis and animal traceability could be performed. Methodology With regards to the reference protocol used in the preparation of ear notch samples, alternative procedures were tested in terms of BVDV analytic sensitivity, diagnostic sensitivity and specificity, as well as quality and purity of animal DNA. Principal Findings/Significance The Allflex DNA Buffer D showed promising results in BVDV diagnosis and genome analyses, opening new perspectives for the livestock industry by the exploitation of the animal genome. Due to the high number of cattle involved in the Belgian official BVDV eradication programme based on ear notch tags sample, a large database on both BVDV status of newborn calves and cattle genome could be created for subsequent different uses (e.g. traceability, determination of parentage, genetic signatures throughout the genome associated with particular traits) evolving through a more integrated animal health. PMID:27764130

  3. Analytical Raman spectroscopic study for discriminant analysis of different animal-derived feedstuff: Understanding the high correlation between Raman spectroscopy and lipid characteristics.

    PubMed

    Gao, Fei; Xu, Lingzhi; Zhang, Yuejing; Yang, Zengling; Han, Lujia; Liu, Xian

    2018-02-01

    The objectives of the current study were to explore the correlation between Raman spectroscopy and lipid characteristics and to assess the potential of Raman spectroscopic methods for distinguishing the different sources of animal-originated feed based on lipid characteristics. A total of 105 lipid samples derived from five animal species have been analyzed by gas chromatography (GC) and FT-Raman spectroscopy. High correlations (r 2 >0.94) were found between the characteristic peak ratio of the Raman spectra (1654/1748 and 1654/1445) and the degree of unsaturation of the animal lipids. The results of FT-Raman data combined with chemometrics showed that the fishmeal, poultry, porcine and ruminant (bovine and ovine) MBMs could be well separated based on their lipid spectral characteristics. This study demonstrated that FT-Raman spectroscopy can mostly exhibit the lipid structure specificity of different species of animal-originated feed and can be used to discriminate different animal-originated feed samples. Copyright © 2017. Published by Elsevier Ltd.

  4. Wildlife monitoring across multiple spatial scales using grid-based sampling

    Treesearch

    Kevin S. McKelvey; Samuel A. Cushman; Michael K. Schwartz; Leonard F. Ruggiero

    2009-01-01

    Recently, noninvasive genetic sampling has become the most effective way to reliably sample occurrence of many species. In addition, genetic data provide a rich data source enabling the monitoring of population status. The combination of genetically based animal data collected at known spatial coordinates with vegetation, topography, and other available covariates...

  5. Authorization of Animal Experiments Is Based on Confidence Rather than Evidence of Scientific Rigor

    PubMed Central

    Nathues, Christina; Würbel, Hanno

    2016-01-01

    Accumulating evidence indicates high risk of bias in preclinical animal research, questioning the scientific validity and reproducibility of published research findings. Systematic reviews found low rates of reporting of measures against risks of bias in the published literature (e.g., randomization, blinding, sample size calculation) and a correlation between low reporting rates and inflated treatment effects. That most animal research undergoes peer review or ethical review would offer the possibility to detect risks of bias at an earlier stage, before the research has been conducted. For example, in Switzerland, animal experiments are licensed based on a detailed description of the study protocol and a harm–benefit analysis. We therefore screened applications for animal experiments submitted to Swiss authorities (n = 1,277) for the rates at which the use of seven basic measures against bias (allocation concealment, blinding, randomization, sample size calculation, inclusion/exclusion criteria, primary outcome variable, and statistical analysis plan) were described and compared them with the reporting rates of the same measures in a representative sub-sample of publications (n = 50) resulting from studies described in these applications. Measures against bias were described at very low rates, ranging on average from 2.4% for statistical analysis plan to 19% for primary outcome variable in applications for animal experiments, and from 0.0% for sample size calculation to 34% for statistical analysis plan in publications from these experiments. Calculating an internal validity score (IVS) based on the proportion of the seven measures against bias, we found a weak positive correlation between the IVS of applications and that of publications (Spearman’s rho = 0.34, p = 0.014), indicating that the rates of description of these measures in applications partly predict their rates of reporting in publications. These results indicate that the authorities licensing animal experiments are lacking important information about experimental conduct that determines the scientific validity of the findings, which may be critical for the weight attributed to the benefit of the research in the harm–benefit analysis. Similar to manuscripts getting accepted for publication despite poor reporting of measures against bias, applications for animal experiments may often be approved based on implicit confidence rather than explicit evidence of scientific rigor. Our findings shed serious doubt on the current authorization procedure for animal experiments, as well as the peer-review process for scientific publications, which in the long run may undermine the credibility of research. Developing existing authorization procedures that are already in place in many countries towards a preregistration system for animal research is one promising way to reform the system. This would not only benefit the scientific validity of findings from animal experiments but also help to avoid unnecessary harm to animals for inconclusive research. PMID:27911892

  6. Authorization of Animal Experiments Is Based on Confidence Rather than Evidence of Scientific Rigor.

    PubMed

    Vogt, Lucile; Reichlin, Thomas S; Nathues, Christina; Würbel, Hanno

    2016-12-01

    Accumulating evidence indicates high risk of bias in preclinical animal research, questioning the scientific validity and reproducibility of published research findings. Systematic reviews found low rates of reporting of measures against risks of bias in the published literature (e.g., randomization, blinding, sample size calculation) and a correlation between low reporting rates and inflated treatment effects. That most animal research undergoes peer review or ethical review would offer the possibility to detect risks of bias at an earlier stage, before the research has been conducted. For example, in Switzerland, animal experiments are licensed based on a detailed description of the study protocol and a harm-benefit analysis. We therefore screened applications for animal experiments submitted to Swiss authorities (n = 1,277) for the rates at which the use of seven basic measures against bias (allocation concealment, blinding, randomization, sample size calculation, inclusion/exclusion criteria, primary outcome variable, and statistical analysis plan) were described and compared them with the reporting rates of the same measures in a representative sub-sample of publications (n = 50) resulting from studies described in these applications. Measures against bias were described at very low rates, ranging on average from 2.4% for statistical analysis plan to 19% for primary outcome variable in applications for animal experiments, and from 0.0% for sample size calculation to 34% for statistical analysis plan in publications from these experiments. Calculating an internal validity score (IVS) based on the proportion of the seven measures against bias, we found a weak positive correlation between the IVS of applications and that of publications (Spearman's rho = 0.34, p = 0.014), indicating that the rates of description of these measures in applications partly predict their rates of reporting in publications. These results indicate that the authorities licensing animal experiments are lacking important information about experimental conduct that determines the scientific validity of the findings, which may be critical for the weight attributed to the benefit of the research in the harm-benefit analysis. Similar to manuscripts getting accepted for publication despite poor reporting of measures against bias, applications for animal experiments may often be approved based on implicit confidence rather than explicit evidence of scientific rigor. Our findings shed serious doubt on the current authorization procedure for animal experiments, as well as the peer-review process for scientific publications, which in the long run may undermine the credibility of research. Developing existing authorization procedures that are already in place in many countries towards a preregistration system for animal research is one promising way to reform the system. This would not only benefit the scientific validity of findings from animal experiments but also help to avoid unnecessary harm to animals for inconclusive research.

  7. Reports of severe physical punishment and exposure to animal cruelty by inmates convicted of felonies and by university students.

    PubMed

    Miller, K S; Knutson, J F

    1997-01-01

    A self-report questionnaire designed to assess abusive childhood environments and exposure to animal cruelty was administered to 314 inmates in a prisoner classification center. Although high rates of physical punishment characterized the entire sample, persons charged with violent, but nonhomicidal crimes reported more severely punitive childhood histories than those charged with homicidal crimes, sex offenses, and nonviolent offenses. Some exposure to animal cruelty was widespread in the sample, but there was no association between experiencing animal cruelty and the type of crime committed. Moreover, there were only modest associations between animal cruelty experiences and the aversive childhood histories of the subjects, as well as the subjects' reported use of physical and sexual coercion in dating and intimate relationships. To determine whether the high base rate of exposure to animal cruelty was unique to the incarcerated sample, a follow-up study was completed with university undergraduates. Widespread exposure to some animal cruelty was reported by undergraduates; there were modest associations between reporting animal cruelty and reporting punitive and acrimonious childhood histories. In general, the findings were consistent with the hypothesis that there is an association between punitive childhood histories and antisocial behavior but not consistent with the hypothesis that exposure to animal cruelty is importantly related to antisocial behavior or child maltreatment.

  8. Extrahuman Epidemiology of Acinetobacter baumannii in Lebanon

    PubMed Central

    Rafei, Rayane; Hamze, Monzer; Pailhoriès, Hélène; Eveillard, Matthieu; Marsollier, Laurent; Joly-Guillou, Marie-Laure; Dabboussi, Fouad

    2015-01-01

    The presence of Acinetobacter baumannii outside hospitals is still a controversial issue. The objective of our study was to explore the extrahospital epidemiology of A. baumannii in Lebanon. From February 2012 to October 2013, a total of 73 water samples, 51 soil samples, 37 raw cow milk samples, 50 cow meat samples, 7 raw cheese samples, and 379 animal samples were analyzed by cultural methods for the presence of A. baumannii. Species identification was performed by rpoB gene sequencing. Antibiotic susceptibility was investigated, and the A. baumannii population was studied by two genotyping approaches: multilocus sequence typing (MLST) and blaOXA-51 sequence-based typing (SBT). A. baumannii was detected in 6.9% of water samples, 2.7% of milk samples, 8.0% of meat samples, 14.3% of cheese samples, and 7.7% of animal samples. All isolates showed a susceptible phenotype against most of the antibiotics tested and lacked carbapenemase-encoding genes, except one that harbored a blaOXA-143 gene. MLST analysis revealed the presence of 36 sequence types (STs), among which 24 were novel STs reported for the first time in this study. blaOXA-51 SBT showed the presence of 34 variants, among which 21 were novel and all were isolated from animal origins. Finally, 30 isolates had new partial rpoB sequences and were considered putative new Acinetobacter species. In conclusion, animals can be a potential reservoir for A. baumannii and the dissemination of new emerging carbapenemases. The roles of the novel animal clones identified in community-acquired infections should be investigated. PMID:25616788

  9. Magnetic Stirrer Method for the Detection of Trichinella Larvae in Muscle Samples.

    PubMed

    Mayer-Scholl, Anne; Pozio, Edoardo; Gayda, Jennifer; Thaben, Nora; Bahn, Peter; Nöckler, Karsten

    2017-03-03

    Trichinellosis is a debilitating disease in humans and is caused by the consumption of raw or undercooked meat of animals infected with the nematode larvae of the genus Trichinella. The most important sources of human infections worldwide are game meat and pork or pork products. In many countries, the prevention of human trichinellosis is based on the identification of infected animals by means of the artificial digestion of muscle samples from susceptible animal carcasses. There are several methods based on the digestion of meat but the magnetic stirrer method is considered the gold standard. This method allows the detection of Trichinella larvae by microscopy after the enzymatic digestion of muscle samples and subsequent filtration and sedimentation steps. Although this method does not require special and expensive equipment, internal controls cannot be used. Therefore, stringent quality management should be applied throughout the test. The aim of the present work is to provide detailed handling instructions and critical control points of the method to analysts, based on the experience of the European Union Reference Laboratory for Parasites and the National Reference Laboratory of Germany for Trichinella.

  10. Ambient Molecular Analysis of Biological Tissue Using Low-Energy, Femtosecond Laser Vaporization and Nanospray Postionization Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Shi, Fengjian; Flanigan, Paul M.; Archer, Jieutonne J.; Levis, Robert J.

    2016-03-01

    Direct analysis of plant and animal tissue samples by laser electrospray mass spectrometry (LEMS) was investigated using low-energy, femtosecond duration laser vaporization at wavelengths of 800 and 1042 nm followed by nanospray postionization. Low-energy (<50 μJ), fiber-based 1042 nm LEMS (F-LEMS) allowed interrogation of the molecular species in fresh flower petal and leaf samples using 435 fs, 10 Hz bursts of 20 pulses from a Ytterbium-doped fiber laser and revealed comparable results to high energy (75-1120 μJ), 45 fs, 800 nm Ti:Sapphire-based LEMS (Ti:Sapphire-LEMS) measurements. Anthocyanins, sugars, and other metabolites were successfully detected and revealed the anticipated metabolite profile for the petal and leaf samples. Phospholipids, especially phosphatidylcholine, were identified from a fresh mouse brain section sample using Ti:Sapphire-LEMS without the application of matrix. These lipid features were suppressed in both the fiber-based and Ti:Sapphire-based LEMS measurements when the brain sample was prepared using the optimal cutting temperature compounds that are commonly used in animal tissue cryosections.

  11. Evaluation of a new serological technique for detecting rabies virus antibodies following vaccination.

    PubMed

    Ma, Xiaoyue; Niezgoda, Michael; Blanton, Jesse D; Recuenco, Sergio; Rupprecht, Charles E

    2012-08-03

    Two major techniques are currently used to estimate rabies virus antibody values: neutralization assays, such as the rapid fluorescent focus inhibition test (RFFIT), and enzyme-linked immunosorbent assays (ELISAs). The RFFIT is considered the gold standard assay and has been used to assess the titer of rabies virus neutralizing antibodies for more than three decades. In the late 1970s, ELISA began to be used to estimate the level of rabies virus antibody and has recently been used by some laboratories as an alternate screening test for animal sera. Although the ELISA appears simpler, safer and more efficient, the assay is less sensitive in detecting low values of rabies virus neutralizing antibodies than neutralization tests. This study was designed to evaluate a new ELISA-based method for detecting rabies virus binding antibody. This new technique uses electro-chemi-luminescence labels and carbon electrode plates to detect binding events. In this comparative study, the RFFIT and the new ELISA-based technique were used to evaluate the level of rabies virus antibodies in human and animal serum samples. By using a conservative approximation of 0.15 IU/ml as a cutoff point, the new ELISA-based technique demonstrated a sensitivity of 100% and a specificity of 95% for human samples and for experimental animal samples. The sensitivity and specificity for field animal samples was 96% and 95%, respectively. The preliminary results from this study appear promising and demonstrate a higher sensitivity than traditional ELISA methods. Published by Elsevier Ltd.

  12. Evaluation of Selected Nutrients and Contaminants in Distillers Grains from Ethanol Production in Texas.

    PubMed

    Lee, Kyung-Min; Herrman, Timothy J; Post, Lynn

    2016-09-01

    This article contains the results of the evaluation of distillers grain (DG) coproducts from different ethanol plants around the United States and supplemented in animal diets in Texas, based on samples analyzed from 2008 to 2014. The samples were assessed for concentration, occurrence, and prevalence of selected nutrients and contaminants. Protein and sulfur contents of DG were largely different between maize and sorghum coproducts, as well as wet distillers grain with solubles (WDGS) and dried distillers grain with solubles (DDGS), indicating a significant effect of grain feedstock and dry grind process stream on DG composition and quality. Salmonella was isolated in 4 DDGS samples of a total of 157 DG samples, a percentage (2.5%) that is lower than the percentage of Salmonella -positive samples found in other feed samples analyzed during the same period. A small amount of virginiamycin residue was found in 24 maize DDGS, 1 maize WDGS, and 2 sorghum DDGS samples of 242 samples in total. One sorghum DDGS sample of 168 DG samples was contaminated with animal protein prohibited for use in ruminant feed and was channeled to poultry feed. The concentrations of aflatoxin and fumonisin DG coproducts averaged 3.4 μg/kg and 0.7 mg/kg, respectively. Among contaminated maize DG samples, five DDGS samples for aflatoxin contained a higher concentration than the U.S. Food and Drug Administration (FDA) minimum action level of 20 μg/kg for use in animal feed, whereas no sample for fumonisin was found above the action level of 5 mg/kg. The study provides the most current results involving DG coproducts and associated hazards that will assist development of food safety plans required by the FDA in their September 2015 rule titled "Current Good Manufacturing Practice Hazard Analysis and Risk Based Preventive Controls for Food for Animals."

  13. User-controlled photographic animations, photograph-based questions, and questionnaires: three Internet-based instruments for measuring drivers' risk-taking behavior.

    PubMed

    Horswill, M S; Coster, M E

    2001-02-01

    The Internet has been exploited successfully in the past as a medium for behavioral research. This paper presents a series of studies designed to assess Internet-based measures of drivers' risk-taking behavior. First, we compared responses from an Internet sample with a traditional pencil-and-paper sample using established questionnaire measures of risk taking. No significant differences were found. Second, we assessed the validity of new Internet-based instruments, involving photographs and photographic animations, that measured speed, gap acceptance, and passing. Responses were found to reflect known demographic patterns of actual behavior to some degree. Also, a roadside survey of speeds was carried out at the locations depicted in the photographic measure of speeding and, with certain exceptions, differences between the two appeared to be constant. Third, a between-subject experimental manipulation involving the photographic animation measure of gap acceptance was used to demonstrate one application of these techniques.

  14. Childhood and adolescent animal cruelty methods and their possible link to adult violent crimes.

    PubMed

    Hensley, Christopher; Tallichet, Suzanne E

    2009-01-01

    Few researchers have investigated the potentially predictive power of childhood and adolescent animal cruelty methods as they are associated with subsequent interpersonal violence in adulthood. Based on a sample of 261 inmates at medium- and maximum-security prisons in a southern state, the present study examines the relationship between several retrospectively reported animal cruelty methods (drowned, hit or kicked, shot, choked, burned, and had sex) and violent criminal acts committed against humans (assault, rape, and murder). More than half of the sample reported they had shot animals, and almost half had either kicked or hit them. About one in five said they had choked animals, and about one in seven said they had either drowned, burned, or had sex with them. Regression analyses revealed that drowning and having sex with an animal was predictive of later interpersonal violence as adults.

  15. Childhood Animal Cruelty Methods and Their Link to Adult Interpersonal Violence

    ERIC Educational Resources Information Center

    Henderson, Brandy B.; Hensley, Christopher; Tallichet, Suzanne E.

    2011-01-01

    Recent research has begun to establish a relationship between childhood acts of animal cruelty and later violence against humans. However, few studies have focused on the influence of animal cruelty methods on later interpersonal violence. In a replication of a study by Hensley and Tallichet (2009) and based on a sample of 180 inmates at medium-…

  16. Nutrient composition, digestible and metabolizable energy content, and prediction of energy for animal protein by-products in finishing pig diets

    USDA-ARS?s Scientific Manuscript database

    An industry survey and an animal experiment were conducted to evaluate compositional variability and DE and ME content of animal protein by-products, and to generate equations to predict DE and ME content based on chemical analysis. For the 220 samples collected, the greatest concentration of CP was...

  17. Estimating clinical chemistry reference values based on an existing data set of unselected animals.

    PubMed

    Dimauro, Corrado; Bonelli, Piero; Nicolussi, Paola; Rassu, Salvatore P G; Cappio-Borlino, Aldo; Pulina, Giuseppe

    2008-11-01

    In an attempt to standardise the determination of biological reference values, the International Federation of Clinical Chemistry (IFCC) has published a series of recommendations on developing reference intervals. The IFCC recommends the use of an a priori sampling of at least 120 healthy individuals. However, such a high number of samples and laboratory analysis is expensive, time-consuming and not always feasible, especially in veterinary medicine. In this paper, an alternative (a posteriori) method is described and is used to determine reference intervals for biochemical parameters of farm animals using an existing laboratory data set. The method used was based on the detection and removal of outliers to obtain a large sample of animals likely to be healthy from the existing data set. This allowed the estimation of reliable reference intervals for biochemical parameters in Sarda dairy sheep. This method may also be useful for the determination of reference intervals for different species, ages and gender.

  18. Using an Animated Case Scenario Based on Constructivist 5E Model to Enhance Pre-Service Teachers' Awareness of Electrical Safety

    ERIC Educational Resources Information Center

    Hirca, Necati

    2013-01-01

    The objective of this study is to get pre-service teachers to develop an awareness of first aid knowledge and skills related to electrical shocking and safety within a scenario based animation based on a Constructivist 5E model. The sample of the study was composed of 78 (46 girls and 32 boys) pre-service classroom teachers from two faculties of…

  19. Ape Conservation Physiology: Fecal Glucocorticoid Responses in Wild Pongo pygmaeus morio following Human Visitation

    PubMed Central

    Muehlenbein, Michael P.; Ancrenaz, Marc; Sakong, Rosman; Ambu, Laurentius; Prall, Sean; Fuller, Grace; Raghanti, Mary Ann

    2012-01-01

    Nature-based tourism can generate important revenue to support conservation of biodiversity. However, constant exposure to tourists and subsequent chronic activation of stress responses can produce pathological effects, including impaired cognition, growth, reproduction, and immunity in the same animals we are interested in protecting. Utilizing fecal samples (N = 53) from 2 wild habituated orangutans (Pongo pygmaeus morio) (in addition to 26 fecal samples from 4 wild unhabituated orangutans) in the Lower Kinabatangan Wildlife Sanctuary of Sabah, Malaysian Borneo, we predicted that i) fecal glucocorticoid metabolite concentrations would be elevated on the day after tourist visitation (indicative of normal stress response to exposure to tourists on the previous day) compared to samples taken before or during tourist visitation in wild, habituated orangutans, and ii) that samples collected from habituated animals would have lower fecal glucocorticoid metabolites than unhabituated animals not used for tourism. Among the habituated animals used for tourism, fecal glucocorticoid metabolite levels were significantly elevated in samples collected the day after tourist visitation (indicative of elevated cortisol production on the previous day during tourist visitation). Fecal glucocorticoid metabolite levels were also lower in the habituated animals compared to their age-matched unhabituated counterparts. We conclude that the habituated animals used for this singular ecotourism project are not chronically stressed, unlike other species/populations with documented permanent alterations in stress responses. Animal temperament, species, the presence of coping/escape mechanisms, social confounders, and variation in amount of tourism may explain differences among previous experiments. Acute alterations in glucocorticoid measures in wildlife exposed to tourism must be interpreted conservatively. While permanently altered stress responses can be detrimental, preliminary results in these wild habituated orangutans suggest that low levels of predictable disturbance can likely result in low physiological impact on these animals. PMID:22438916

  20. Ape conservation physiology: fecal glucocorticoid responses in wild Pongo pygmaeus morio following human visitation.

    PubMed

    Muehlenbein, Michael P; Ancrenaz, Marc; Sakong, Rosman; Ambu, Laurentius; Prall, Sean; Fuller, Grace; Raghanti, Mary Ann

    2012-01-01

    Nature-based tourism can generate important revenue to support conservation of biodiversity. However, constant exposure to tourists and subsequent chronic activation of stress responses can produce pathological effects, including impaired cognition, growth, reproduction, and immunity in the same animals we are interested in protecting. Utilizing fecal samples (N = 53) from 2 wild habituated orangutans (Pongo pygmaeus morio) (in addition to 26 fecal samples from 4 wild unhabituated orangutans) in the Lower Kinabatangan Wildlife Sanctuary of Sabah, Malaysian Borneo, we predicted that i) fecal glucocorticoid metabolite concentrations would be elevated on the day after tourist visitation (indicative of normal stress response to exposure to tourists on the previous day) compared to samples taken before or during tourist visitation in wild, habituated orangutans, and ii) that samples collected from habituated animals would have lower fecal glucocorticoid metabolites than unhabituated animals not used for tourism. Among the habituated animals used for tourism, fecal glucocorticoid metabolite levels were significantly elevated in samples collected the day after tourist visitation (indicative of elevated cortisol production on the previous day during tourist visitation). Fecal glucocorticoid metabolite levels were also lower in the habituated animals compared to their age-matched unhabituated counterparts. We conclude that the habituated animals used for this singular ecotourism project are not chronically stressed, unlike other species/populations with documented permanent alterations in stress responses. Animal temperament, species, the presence of coping/escape mechanisms, social confounders, and variation in amount of tourism may explain differences among previous experiments. Acute alterations in glucocorticoid measures in wildlife exposed to tourism must be interpreted conservatively. While permanently altered stress responses can be detrimental, preliminary results in these wild habituated orangutans suggest that low levels of predictable disturbance can likely result in low physiological impact on these animals.

  1. Didelphis marsupialis (common opossum): a potential reservoir host for zoonotic leishmaniasis in the metropolitan region of Belo Horizonte (Minas Gerais, Brazil).

    PubMed

    Schallig, Henk D F H; da Silva, Eduardo S; van der Meide, Wendy F; Schoone, Gerard J; Gontijo, Celia M F

    2007-01-01

    Identification of the zoonotic reservoir is important for leishmaniasis control program. A number of (wild) animal species may serve as reservoir hosts, including the opossum Didelphis marsupialis. A survey carried out in Didelphis specimens (n = 111) from the metropolitan region of Belo Horizonte, an important focus of human leishmaniasis in Brazil, is reported. All animals were serologically tested with indirect fluorescence antibody test (IFAT) and direct agglutination tests (DAT) based on L. (L.) donovani or L. (V.) braziliensis antigen. A sub-population (n = 20) was analyzed with polymerase chain reaction (PCR) for the presence of Leishmania-specific DNA. For species identification, PCR-positive samples were subjected to restriction enzyme fragment polymorphism (RFLP) analysis. Depending on the sero-diagnostic test employed, the sero-prevalence varied between 8.1% (9/111 animals positive with DAT test based on L. braziliensis antigen) and 21.6% (24/111 animals positive with IFAT). Five out of 20 samples analyzed with PCR tested positive for the presence of Leishmania-specific DNA. RFLP analysis revealed that two samples contained L. braziliensis complex DNA, one contained L. donovani complex DNA, and two samples could not be typed with the methodology used. These data suggest a potential role for the opossum as a reservoir host for zoonotic leishmaniasis in the region.

  2. Evidence of Mycobacterium tuberculosis complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA.

    PubMed

    Swift, Benjamin M C; Convery, Thomas W; Rees, Catherine E D

    2016-10-02

    Bovine tuberculosis is a zoonotic infectious disease caused by Mycobacterium bovis that affects cattle and can cause tuberculosis in a range of wildlife animals. A bacteriophage-based method combined with PCR (phage-PCR) has been recently used to detect and identify viable pathogenic mycobacteria in the peripheral blood mononuclear cells (PBMCs) of animals suffering from paratuberculosis. To adapt this method for the detection of M. bovis in blood, a new isothermal DNA amplification protocol using Recombinase Polymerase Amplification (RPA) was developed and was found to be able to detect M. bovis BCG within 48 h, with a limit of detection of approximately 10 cells per ml of blood for artificially inoculated blood samples. When blood samples (2 ml) from a Single Comparative Cervical Intradermal Tuberculin (SCCIT)- negative beef herd were tested, Mycobacterium tuberculosis complex (MTC) cells were not detected from any (45) of the blood samples. However when blood samples from SCCIT-positive animals were tested, viable MTC bacteria were detected in 66 % (27/41) of samples. Of these 41 animals sampled, 32 % (13) had visible lesions. In the visible lesion (VL) group, 85 % (11/13) had detectable levels of MTC whereas only 57 % (16/28) of animals which had no visible lesions (NVL) were found to have detectable mycobacteraemia. These results indicated that this simple, rapid method can be applied for the study of M. bovis infections. The frequency with which viable mycobacteria were detected in the peripheral blood of SCCIT-positive animals changes the paradigm of this disease.

  3. A System Overview of the Electronic Surveillance System for the Early Notification of Community-based Epidemics

    DTIC Science & Technology

    2004-11-15

    Doctors’ Visits Animal Health Diagnostic Labs Hospital Emergency Room Local Military & Civ. Users Public Health Surveillance & Epidemiology...Environmental Samples EPA Over the Counter Sales Animal Health Absenteeism ESSENCE I NCR Military Data Tri-Service Outpatient Visits Pharmacy Data

  4. A novel approach to quantifying the spatiotemporal behavior of instrumented grey seals used to sample the environment.

    PubMed

    Baker, Laurie L; Mills Flemming, Joanna E; Jonsen, Ian D; Lidgard, Damian C; Iverson, Sara J; Bowen, W Don

    2015-01-01

    Paired with satellite location telemetry, animal-borne instruments can collect spatiotemporal data describing the animal's movement and environment at a scale relevant to its behavior. Ecologists have developed methods for identifying the area(s) used by an animal (e.g., home range) and those used most intensely (utilization distribution) based on location data. However, few have extended these models beyond their traditional roles as descriptive 2D summaries of point data. Here we demonstrate how the home range method, T-LoCoH, can be expanded to quantify collective sampling coverage by multiple instrumented animals using grey seals (Halichoerus grypus) equipped with GPS tags and acoustic transceivers on the Scotian Shelf (Atlantic Canada) as a case study. At the individual level, we illustrate how time and space-use metrics quantifying individual sampling coverage may be used to determine the rate of acoustic transmissions received. Grey seals collectively sampled an area of 11,308 km (2) and intensely sampled an area of 31 km (2) from June-December. The largest area sampled was in July (2094.56 km (2)) and the smallest area sampled occurred in August (1259.80 km (2)), with changes in sampling coverage observed through time. T-LoCoH provides an effective means to quantify changes in collective sampling effort by multiple instrumented animals and to compare these changes across time. We also illustrate how time and space-use metrics of individual instrumented seal movement calculated using T-LoCoH can be used to account for differences in the amount of time a bioprobe (biological sampling platform) spends in an area.

  5. Dense mesh sampling for video-based facial animation

    NASA Astrophysics Data System (ADS)

    Peszor, Damian; Wojciechowska, Marzena

    2016-06-01

    The paper describes an approach for selection of feature points on three-dimensional, triangle mesh obtained using various techniques from several video footages. This approach has a dual purpose. First, it allows to minimize the data stored for the purpose of facial animation, so that instead of storing position of each vertex in each frame, one could store only a small subset of vertices for each frame and calculate positions of others based on the subset. Second purpose is to select feature points that could be used for anthropometry-based retargeting of recorded mimicry to another model, with sampling density beyond that which can be achieved using marker-based performance capture techniques. Developed approach was successfully tested on artificial models, models constructed using structured light scanner, and models constructed from video footages using stereophotogrammetry.

  6. Modeling misidentification errors that result from use of genetic tags in capture-recapture studies

    USGS Publications Warehouse

    Yoshizaki, J.; Brownie, C.; Pollock, K.H.; Link, W.A.

    2011-01-01

    Misidentification of animals is potentially important when naturally existing features (natural tags) such as DNA fingerprints (genetic tags) are used to identify individual animals. For example, when misidentification leads to multiple identities being assigned to an animal, traditional estimators tend to overestimate population size. Accounting for misidentification in capture-recapture models requires detailed understanding of the mechanism. Using genetic tags as an example, we outline a framework for modeling the effect of misidentification in closed population studies when individual identification is based on natural tags that are consistent over time (non-evolving natural tags). We first assume a single sample is obtained per animal for each capture event, and then generalize to the case where multiple samples (such as hair or scat samples) are collected per animal per capture occasion. We introduce methods for estimating population size and, using a simulation study, we show that our new estimators perform well for cases with moderately high capture probabilities or high misidentification rates. In contrast, conventional estimators can seriously overestimate population size when errors due to misidentification are ignored. ?? 2009 Springer Science+Business Media, LLC.

  7. Effects of diet composition on mutagenic activity in urine.

    PubMed

    Ohara, Akihiro; Matsuhisa, Tsugio

    2004-01-01

    The effects of dietary habits on mutagenic activity in urine were investigated using the umu test based on the use of the genetically engineered bacteria Salmonella typhimurium TA 1535 pSK1002. Genotoxic effects in sample urine were detected by measuring the activation of the SOS response in the bacteria and recording the beta- galactosidase activity. Human subjects consisted of smokers and non-smokers. Urine from subjects who consumed fish showed the highest mutagenic activity, followed by the urine samples from subjects who ate pork or beef. Chicken induced a low level of mutagenic activity. When the subjects ate fried or roasted animal foods, the urine samples gave higher mutagenicity than the urine samples from the subject who consumed non-fried or non-roasted animal foods. When the subject ate vegetables along with a diet rich in animal foods, the activity in urine decreased. Herbs and spices gave the same tendency toward decline as vegetables. Non-smoker urine shower mutagenic activity than samples from smokers.

  8. [Identification of Animal Whole Blood Based on Near Infrared Transmission Spectroscopy].

    PubMed

    Wan, Xiong; Wang, Jian; Liu, Peng-xi; Zhang, Ting-ting

    2016-01-01

    The inspection and classification for blood products are important but complicated in import-export ports or inspection and quarantine departments. For the inspection of whole blood products, open sampling can cause pollution and virulence factors in bloods samples may even endanger inspectors. Thus non-contact classification and identification methods for whole bloods of animals are needed. Spectroscopic techniques adopted in the flowcytometry need sampling blood cells during the detection; therefore they can not meet the demand of non-contact identification and classification for whole bloods of animals. Infrared absorption spectroscopy is a technique that can be used to analyze the molecular structure and chemical bonds of detected samples under the condition of non-contact. To find a feasible spectroscopic approach of non-contact detection for the species variation in whole blood samples, a near infrared transmitted spectra (NITS, 4 497.669 - 7 506.4 cm(-1)) experiment of whole blood samples of three common animals including chickens, dogs and cats has been conducted. During the experiment, the spectroscopic resolution is 5 cm(-1), and each spectrogram is an average of 5 measured spectral data. Experimental results show that all samples have a sharp absorption peak between 5 184 and 5 215 cm(-1), and a gentle absorption peak near 7 000 cm(-1). Besides, the NITS curves of different samples of same animals are similar, and only have slight differences in the whole transmittance. A correlation coefficient (CC) is induced to distinguish the differences of the three animals' whole bloods in NITS curves, and the computed CCs between NITS curves of different samples of the same animals, are greater than 0.99, whereas CCs between NITS curves of the whole bloods of different animals are from 0.509 48 to 0.916 13. Among which CCs between NITS curves of the whole bloods of chickens and cats are from 0.857 23 to 0.912 44, CCs between NITS curves of the whole bloods of chickens and dogs are from 0.509 48 to 0.664 82, and CCs between NITS curves of the whole bloods of cats and dogs are from 0.872 75 to 0.916 13. The cat and the dog belong to the class of mammal, and the CCs between their whole bloods NITS curves are greater than those between chickens and cats, or chickens and dogs, which are hetero-class animals. Namely, the whole bloods NITS curves of the cat and the dog have higher similarity. These results of NITS provide a feasible method of non-contact identification of animal whole bloods.

  9. A quantum dot-based immunoassay for screening of tylosin and tilmicosin in edible animal tissues.

    PubMed

    Le, Tao; Zhu, Liqian; Yang, Xian

    2015-01-01

    A rapid, indirect competitive fluorescence-linked immunosorbent assay (ic-FLISA) based on quantum dots (QDs) as the fluorescent marker was developed for the detection of tylosin and tilmicosin in edible animal tissues. The end point fluorescent detection system was carried out using QDs conjugated with goat anti-mouse secondary antibody. The limits of detection (LODs) for the determination of tylosin and tilmicosin were 0.02 and 0.04 μg kg(-1), respectively. This detection method was used to analyse spiked samples and the recoveries ranged from 83.5% to 98.7% for tylosin and from 81.8% to 98.2% for tilmicosin. In real porcine tissue sample analysis, the results of ic-FLISA were similar to those obtained from an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) to an HPLC method indicating its potential for tylosin and tilmicosin screening in edible animal tissues.

  10. A Template Analysis of Intimate Partner Violence Survivors' Experiences of Animal Maltreatment: Implications for Safety Planning and Intervention.

    PubMed

    Collins, Elizabeth A; Cody, Anna M; McDonald, Shelby Elaine; Nicotera, Nicole; Ascione, Frank R; Williams, James Herbert

    2018-03-01

    This study explores the intersection of intimate partner violence (IPV) and animal cruelty in an ethnically diverse sample of 103 pet-owning IPV survivors recruited from community-based domestic violence programs. Template analysis revealed five themes: (a) Animal Maltreatment by Partner as a Tactic of Coercive Power and Control, (b) Animal Maltreatment by Partner as Discipline or Punishment of Pet, (c) Animal Maltreatment by Children, (d) Emotional and Psychological Impact of Animal Maltreatment Exposure, and (e) Pets as an Obstacle to Effective Safety Planning. Results demonstrate the potential impact of animal maltreatment exposure on women and child IPV survivors' health and safety.

  11. A Template Analysis of Intimate Partner Violence Survivors’ Experiences of Animal Maltreatment: Implications for Safety Planning and Intervention

    PubMed Central

    Collins, Elizabeth A.; Cody, Anna M.; McDonald, Shelby Elaine; Nicotera, Nicole; Ascione, Frank R.; Williams, James Herbert

    2018-01-01

    This study explores the intersection of intimate partner violence (IPV) and animal cruelty in an ethnically diverse sample of 103 pet-owning IPV survivors recruited from community-based domestic violence programs. Template analysis revealed five themes: (a) Animal Maltreatment by Partner as a Tactic of Coercive Power and Control, (b) Animal Maltreatment by Partner as Discipline or Punishment of Pet, (c) Animal Maltreatment by Children, (d) Emotional and Psychological Impact of Animal Maltreatment Exposure, and (e) Pets as an Obstacle to Effective Safety Planning. Results demonstrate the potential impact of animal maltreatment exposure on women and child IPV survivors’ health and safety. PMID:29332521

  12. Analysis of sulfonamides, tilmicosin and avermectins residues in typical animal matrices with multi-plug filtration cleanup by liquid chromatography-tandem mass spectrometry detection.

    PubMed

    Qin, Yuhong; Jatamunua, Freedom; Zhang, Jingru; Li, Yanjie; Han, Yongtao; Zou, Nan; Shan, Jihao; Jiang, Yanbin; Pan, Canping

    2017-05-15

    The frequent use of various veterinary drugs could lead to residue bioaccumulation in animal tissues, which could cause dietary risks to human health. In order to quickly analyze the residues, a liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for detecting Sulfonamides, Tilmicosin and Avermectins (AVMs) residues in animal samples. For sample preparation, modified QuEChERS (quick, easy, cheap, effective, rugged and safe) and ultrasound-assisted extraction (UAE) methods were used. For sample cleanup, n-Hexane delipidation and multi-plug filtration cleanup (m-PFC) method based on primary-secondary amine (PSA) and octadecyl-silica (C18) were used, followed by LC-MS/MS analysis. It was validated on 7 animal matrices (bovine, caprine, swine meat and their kidneys, milk) at two fortified concentration levels of 5 and 100μg/kg. The recoveries ranged from 82 to 107% for all analytes with relative standard deviations (RSDs) less than 15%. Matrix-matched calibrations were performed with coefficients of determination above 0.998 for all analytes within concentration levels of 5-500μg/kg. The developed method was successfully used to analysis veterinary drugs of real animal samples from local markets. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Testing the new animal phylogeny: a phylum level molecular analysis of the animal kingdom.

    PubMed

    Bourlat, Sarah J; Nielsen, Claus; Economou, Andrew D; Telford, Maximilian J

    2008-10-01

    The new animal phylogeny inferred from ribosomal genes some years ago has prompted a number of radical rearrangements of the traditional, morphology based metazoan tree. The two main bilaterian clades, Deuterostomia and Protostomia, find strong support, but the protostomes consist of two sister groups, Ecdysozoa and Lophotrochozoa, not seen in morphology based trees. Although widely accepted, not all recent molecular phylogenetic analyses have supported the tripartite structure of the new animal phylogeny. Furthermore, even if the small ribosomal subunit (SSU) based phylogeny is correct, there is a frustrating lack of resolution of relationships between the phyla that make up the three clades of this tree. To address this issue, we have assembled a dataset including a large number of aligned sequence positions as well as a broad sampling of metazoan phyla. Our dataset consists of sequence data from ribosomal and mitochondrial genes combined with new data from protein coding genes (5139 amino acid and 3524 nucleotide positions in total) from 37 representative taxa sampled across the Metazoa. Our data show strong support for the basic structure of the new animal phylogeny as well as for the Mandibulata including Myriapoda. We also provide some resolution within the Lophotrochozoa, where we confirm support for a monophyletic clade of Echiura, Sipuncula and Annelida and surprising evidence of a close relationship between Brachiopoda and Nemertea.

  14. Identification and typing of Brucella spp. in stranded harbour porpoises (Phocoena phocoena) on the Dutch coast.

    PubMed

    Maio, Elisa; Begeman, Lineke; Bisselink, Yvette; van Tulden, Peter; Wiersma, Lidewij; Hiemstra, Sjoukje; Ruuls, Robin; Gröne, Andrea; Roest, Hendrik-Ido-Jan; Willemsen, Peter; van der Giessen, Joke

    2014-09-17

    The presence of Brucella (B.) spp. in harbour porpoises stranded between 2008 and 2011 along the Dutch coast was studied. A selection of 265 tissue samples from 112 animals was analysed using conventional and molecular methods. In total, 4.5% (5/112) of the animals corresponding with 2.3% (6/265) Brucella positive tissue samples were Brucella positive by culture and these were all confirmed by real-time polymerase chain reaction (real-time PCR) based on the insertion element 711 (IS711). In addition, two more Brucella-positive tissue samples from two animals collected in 2011 were identified using real-time PCR resulting in an overall Brucella prevalence of 6.3% (7/112 animals). Brucella spp. were obtained from lungs (n=3), pulmonary lymph node (n=3) and lungworms (n=2). Multi Locus Variable Number of Tandem Repeats (VNTR) Analysis (MLVA) typing based on the MLVA-16 showed that the Brucella isolates were B. ceti. Additional in silico Multi Locus Sequence typing (MLST) after whole genome sequencing of the 6 Brucella isolates confirmed B. ceti ST 23. According to the Brucella 2010 MLVA database, the isolated Brucella strains encountered were of five genotypes, in two distinct subclusters divided in two different time periods of harbour porpoises collection. This study is the first population based analyses for Brucella spp. infections in cetaceans stranded along the Dutch coast. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  15. Constraints to estimating the prevalence of trypanosome infections in East African zebu cattle.

    PubMed

    Cox, Andrew P; Tosas, Olga; Tilley, Aimee; Picozzi, Kim; Coleman, Paul; Hide, Geoff; Welburn, Susan C

    2010-09-06

    In East Africa, animal trypanosomiasis is caused by many tsetse transmitted protozoan parasites including Trypanosoma vivax, T. congolense and subspecies of T. brucei s.l. (T. b. brucei and zoonotic human infective T. b. rhodesiense) that may co-circulate in domestic and wild animals. Accurate species-specific prevalence measurements of these parasites in animal populations are complicated by mixed infections of trypanosomes within individual hosts, low parasite densities and difficulties in conducting field studies. Many Polymerase Chain Reaction (PCR) based diagnostic tools are available to characterise and quantify infection in animals. These are important for assessing the contribution of infections in animal reservoirs and the risk posed to humans from zoonotic trypanosome species. New matrices for DNA capture have simplified large scale field PCR analyses but few studies have examined the impact of these techniques on prevalence estimations. The Whatman FTA matrix has been evaluated using a random sample of 35 village zebu cattle from a population naturally exposed to trypanosome infection. Using a generic trypanosome-specific PCR, prevalence was systematically evaluated. Multiple PCR samples taken from single FTA cards demonstrated that a single punch from an FTA card is not sufficient to confirm the infectivity status of an individual animal as parasite DNA is unevenly distributed across the card. At low parasite densities in the host, this stochastic sampling effect results in underestimation of prevalence based on single punch PCR testing. Repeated testing increased the estimated prevalence of all Trypanosoma spp. from 9.7% to 86%. Using repeat testing, a very high prevalence of pathogenic trypanosomes was detected in these local village cattle: T. brucei (34.3%), T. congolense (42.9%) and T. vivax (22.9%). These results show that, despite the convenience of Whatman FTA cards and specific PCR based detection tools, the chronically low parasitaemias in indigenous African zebu cattle make it difficult to establish true prevalence. Although this study specifically applies to FTA cards, a similar effect would be experienced with other approaches using blood samples containing low parasite densities. For example, using blood film microscopy or PCR detection from liquid samples where the probability of detecting a parasite or DNA molecule, in the required number of fields of view or PCR reaction, is less than one.

  16. Constraints to estimating the prevalence of trypanosome infections in East African zebu cattle

    PubMed Central

    2010-01-01

    Background In East Africa, animal trypanosomiasis is caused by many tsetse transmitted protozoan parasites including Trypanosoma vivax, T. congolense and subspecies of T. brucei s.l. (T. b. brucei and zoonotic human infective T. b. rhodesiense) that may co-circulate in domestic and wild animals. Accurate species-specific prevalence measurements of these parasites in animal populations are complicated by mixed infections of trypanosomes within individual hosts, low parasite densities and difficulties in conducting field studies. Many Polymerase Chain Reaction (PCR) based diagnostic tools are available to characterise and quantify infection in animals. These are important for assessing the contribution of infections in animal reservoirs and the risk posed to humans from zoonotic trypanosome species. New matrices for DNA capture have simplified large scale field PCR analyses but few studies have examined the impact of these techniques on prevalence estimations. Results The Whatman FTA matrix has been evaluated using a random sample of 35 village zebu cattle from a population naturally exposed to trypanosome infection. Using a generic trypanosome-specific PCR, prevalence was systematically evaluated. Multiple PCR samples taken from single FTA cards demonstrated that a single punch from an FTA card is not sufficient to confirm the infectivity status of an individual animal as parasite DNA is unevenly distributed across the card. At low parasite densities in the host, this stochastic sampling effect results in underestimation of prevalence based on single punch PCR testing. Repeated testing increased the estimated prevalence of all Trypanosoma spp. from 9.7% to 86%. Using repeat testing, a very high prevalence of pathogenic trypanosomes was detected in these local village cattle: T. brucei (34.3%), T. congolense (42.9%) and T. vivax (22.9%). Conclusions These results show that, despite the convenience of Whatman FTA cards and specific PCR based detection tools, the chronically low parasitaemias in indigenous African zebu cattle make it difficult to establish true prevalence. Although this study specifically applies to FTA cards, a similar effect would be experienced with other approaches using blood samples containing low parasite densities. For example, using blood film microscopy or PCR detection from liquid samples where the probability of detecting a parasite or DNA molecule, in the required number of fields of view or PCR reaction, is less than one. PMID:20815940

  17. Comparison of Sewage and Animal Fecal Microbiomes by Using Oligotyping Reveals Potential Human Fecal Indicators in Multiple Taxonomic Groups

    PubMed Central

    Fisher, Jenny C.; Eren, A. Murat; Green, Hyatt C.; Shanks, Orin C.; Morrison, Hilary G.; Vineis, Joseph H.; Sogin, Mitchell L.

    2015-01-01

    Most DNA-based microbial source tracking (MST) approaches target host-associated organisms within the order Bacteroidales, but the gut microbiota of humans and other animals contain organisms from an array of other taxonomic groups that might provide indicators of fecal pollution sources. To discern between human and nonhuman fecal sources, we compared the V6 regions of the 16S rRNA genes detected in fecal samples from six animal hosts to those found in sewage (as a proxy for humans). We focused on 10 abundant genera and used oligotyping, which can detect subtle differences between rRNA gene sequences from ecologically distinct organisms. Our analysis showed clear patterns of differential oligotype distributions between sewage and animal samples. Over 100 oligotypes of human origin occurred preferentially in sewage samples, and 99 human oligotypes were sewage specific. Sequences represented by the sewage-specific oligotypes can be used individually for development of PCR-based assays or together with the oligotypes preferentially associated with sewage to implement a signature-based approach. Analysis of sewage from Spain and Brazil showed that the sewage-specific oligotypes identified in U.S. sewage have the potential to be used as global alternative indicators of human fecal pollution. Environmental samples with evidence of prior human fecal contamination had consistent ratios of sewage signature oligotypes that corresponded to the trends observed for sewage. Our methodology represents a promising approach to identifying new bacterial taxa for MST applications and further highlights the potential of the family Lachnospiraceae to provide human-specific markers. In addition to source tracking applications, the patterns of the fine-scale population structure within fecal taxa suggest a fundamental relationship between bacteria and their hosts. PMID:26231648

  18. Rapid determination of trace copper in animal feed based on micro-plate colorimetric reaction and statistical partitioning correction.

    PubMed

    Niu, Yiming; Wang, Jiayi; Zhang, Chi; Chen, Yiqiang

    2017-04-15

    The objective of this study was to develop a micro-plate based colorimetric assay for rapid and high-throughput detection of copper in animal feed. Copper ion in animal feed was extracted by trichloroacetic acid solution and reduced to cuprous ion by hydroxylamine. The cuprous ion can chelate with 2,2'-bicinchoninic acid to form a Cu-BCA complex which was detected with high sensitivity by micro-plate reader at 354nm. The whole assay procedure can be completed within 20min. To eliminate matrix interference, a statistical partitioning correction approach was proposed, which makes the detection of copper in complex samples possible. The limit of detection was 0.035μg/mL and the detection range was 0.1-10μg/mL of copper in buffer solution. Actual sample analysis indicated that this colorimetric assay produced results consistent with atomic absorption spectrometry analysis. These results demonstrated that the developed assay can be used for rapid determination of copper in animal feed. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. ANIMAL BEHAVIOR AND WELL-BEING SYMPOSIUM: The Common Swine Industry Audit: Future steps to assure positive on-farm animal welfare utilizing validated, repeatable and feasible animal-based measures.

    PubMed

    Pairis-Garcia, M; Moeller, S J

    2017-03-01

    The Common Swine Industry Audit (CSIA) was developed and scientifically evaluated through the combined efforts of a task force consisting of university scientists, veterinarians, pork producers, packers, processers, and retail and food service personnel to provide stakeholders throughout the pork chain with a consistent, reliable, and verifiable system to ensure on-farm swine welfare and food safety. The CSIA tool was built from the framework of the Pork Quality Assurance Plus (PQA Plus) site assessment program with the purpose of developing a single, common audit platform for the U.S. swine industry. Twenty-seven key aspects of swine care are captured and evaluated in CSIA and cover the specific focal areas of animal records, animal observations, facilities, and caretakers. Animal-based measures represent approximately 50% of CSIA evaluation criteria and encompass critical failure criteria, including observation of willful acts of abuse and determination of timely euthanasia. Objective, science-based measures of animal well-being parameters (e.g., BCS, lameness, lesions, hernias) are assessed within CSIA using statistically validated sample sizes providing a detection ability of 1% with 95% confidence. The common CSIA platform is used to identify care issues and facilitate continuous improvement in animal care through a validated, repeatable, and feasible animal-based audit process. Task force members provide continual updates to the CSIA tool with a specific focus toward 1) identification and interpretation of appropriate animal-based measures that provide inherent value to pig welfare, 2) establishment of acceptability thresholds for animal-based measures, and 3) interpretation of CSIA data for use and improvement of welfare within the U.S. swine industry.

  20. Sampling guidelines for oral fluid-based surveys of group-housed animals.

    PubMed

    Rotolo, Marisa L; Sun, Yaxuan; Wang, Chong; Giménez-Lirola, Luis; Baum, David H; Gauger, Phillip C; Harmon, Karen M; Hoogland, Marlin; Main, Rodger; Zimmerman, Jeffrey J

    2017-09-01

    Formulas and software for calculating sample size for surveys based on individual animal samples are readily available. However, sample size formulas are not available for oral fluids and other aggregate samples that are increasingly used in production settings. Therefore, the objective of this study was to develop sampling guidelines for oral fluid-based porcine reproductive and respiratory syndrome virus (PRRSV) surveys in commercial swine farms. Oral fluid samples were collected in 9 weekly samplings from all pens in 3 barns on one production site beginning shortly after placement of weaned pigs. Samples (n=972) were tested by real-time reverse-transcription PCR (RT-rtPCR) and the binary results analyzed using a piecewise exponential survival model for interval-censored, time-to-event data with misclassification. Thereafter, simulation studies were used to study the barn-level probability of PRRSV detection as a function of sample size, sample allocation (simple random sampling vs fixed spatial sampling), assay diagnostic sensitivity and specificity, and pen-level prevalence. These studies provided estimates of the probability of detection by sample size and within-barn prevalence. Detection using fixed spatial sampling was as good as, or better than, simple random sampling. Sampling multiple barns on a site increased the probability of detection with the number of barns sampled. These results are relevant to PRRSV control or elimination projects at the herd, regional, or national levels, but the results are also broadly applicable to contagious pathogens of swine for which oral fluid tests of equivalent performance are available. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Estimating Animal Abundance in Ground Beef Batches Assayed with Molecular Markers

    PubMed Central

    Hu, Xin-Sheng; Simila, Janika; Platz, Sindey Schueler; Moore, Stephen S.; Plastow, Graham; Meghen, Ciaran N.

    2012-01-01

    Estimating animal abundance in industrial scale batches of ground meat is important for mapping meat products through the manufacturing process and for effectively tracing the finished product during a food safety recall. The processing of ground beef involves a potentially large number of animals from diverse sources in a single product batch, which produces a high heterogeneity in capture probability. In order to estimate animal abundance through DNA profiling of ground beef constituents, two parameter-based statistical models were developed for incidence data. Simulations were applied to evaluate the maximum likelihood estimate (MLE) of a joint likelihood function from multiple surveys, showing superiority in the presence of high capture heterogeneity with small sample sizes, or comparable estimation in the presence of low capture heterogeneity with a large sample size when compared to other existing models. Our model employs the full information on the pattern of the capture-recapture frequencies from multiple samples. We applied the proposed models to estimate animal abundance in six manufacturing beef batches, genotyped using 30 single nucleotide polymorphism (SNP) markers, from a large scale beef grinding facility. Results show that between 411∼1367 animals were present in six manufacturing beef batches. These estimates are informative as a reference for improving recall processes and tracing finished meat products back to source. PMID:22479559

  2. Induction of parturition with prostaglandin f2 alpha as a possible model to study impaired reproductive performance in the dairy cow.

    PubMed

    Kask, K; Gustafsson, H; Gunnarsson, A; Kindahl, H

    2000-05-31

    Parturitions were induced in five cows, 2 weeks before term using prostaglandin (PG) F(2alpha). Two i.m. injections were performed with an interval of 24 h. All cows calved within 5 days (average 2.7 days) after the first injection of PGF(2alpha). Out of five cows, four had retained fetal membranes (RFM). Each animal was sampled for bacteriological examination using uterine biopsies twice a week during 42 days postpartum (PP). Jugular vein blood samples were withdrawn for PGF(2alpha)-metabolite and progesterone analyses five times per day during the first week PP and eight times per 24 h during the 2nd and 3rd weeks PP. From the 4th week, the sampling interval was reduced back to five times per day. From the 5th week PP, the sampling was reduced to two times per day and sampling was terminated after day 46 PP. Only morning samples were used for progesterone analyses. From day 10 PP, ultrasonography (US) was performed every 3rd day until day 39 PP for detection of ovarian activity and follicular dynamics. The highest incidence of bacteriological species was found during the first 3 weeks PP. After the 5th week of collection, all animals were free from bacteria. The species of bacteria found were Arcanobacterium (Actinomyces) pyogenes, Escherichia coli, alpha-hemolytic streptococcae and Pasteurella multocida. Immediately after parturition, very high levels of the PG-metabolite were seen in all animals, with a sharp decrease to line of significance around days 9-12 PP. Small increases above the line of significance were detected up to day 27 PP in cows with RFM, and after that time the levels were considered to be at baseline. Low levels of progesterone were seen in four animals during the whole experimental time. In one animal, an increase was seen on day 43 PP, which was maintained until the end of the experimental period on day 46 PP. Based on US, follicular waves were detected in all animals during the experimental period. In three animals, three non-ovulatory follicular waves were detected and in two animals, four non-ovulatory follicular waves were detected during 39 days of ultrasound sessions. Based on progesterone levels, only one animal was considered to have ovulated around day 40 PP. Results from the present study indicate that reproductive performance of cows after PG-induced parturitions differs from those of spontaneous cases of RFM. Differences regarding the resumption of ovarian activity were also observed between previous studies of dexamethasone-induced parturitions and the present study.

  3. Evidence of Borna disease virus genome detection in French domestic animals and in foxes (Vulpes vulpes).

    PubMed

    Dauphin, G; Legay, V; Sailleau, C; Smondack, S; Hammoumi, S; Zientara, S

    2001-09-01

    Borna disease virus (BDV) is an enveloped, non-segmented negative-stranded RNA virus which belongs to the Bornaviridae family. BDV is an aetiological agent of encephalitis in horses, sheep and several other vertebrate species. In order to extend our knowledge about the presence of BDV in France, a study based on BDV RNA detection by RT-nested-PCR was done with 196 animal tissues: 171 brain samples collected from different animal species (75 horses, 59 foxes, 31 cattle, 4 dogs, 1 sheep, 1 roe deer) and 25 horse blood samples. An RNA internal standard molecule was constructed and was co-amplified with the test template. This study reports the first detection of BDV RNA in France in 10 brain samples collected from horses, foxes and cattle, and from 14 horse blood samples. Detection of the BDV genome in the brains of six red foxes is the first evidence of BDV infection in this species.

  4. Autocorrelation of location estimates and the analysis of radiotracking data

    USGS Publications Warehouse

    Otis, D.L.; White, Gary C.

    1999-01-01

    The wildlife literature has been contradictory about the importance of autocorrelation in radiotracking data used for home range estimation and hypothesis tests of habitat selection. By definition, the concept of a home range involves autocorrelated movements, but estimates or hypothesis tests based on sampling designs that predefine a time frame of interest, and that generate representative samples of an animal's movement during this time frame, should not be affected by length of the sampling interval and autocorrelation. Intensive sampling of the individual's home range and habitat use during the time frame of the study leads to improved estimates for the individual, but use of location estimates as the sample unit to compare across animals is pseudoreplication. We therefore recommend against use of habitat selection analysis techniques that use locations instead of individuals as the sample unit. We offer a general outline for sampling designs for radiotracking studies.

  5. Evaluating internal and external markers versus fecal sampling procedure interactions when estimating intake in dairy cows consuming a corn silage-based diet.

    PubMed

    Velásquez, A V; da Silva, G G; Sousa, D O; Oliveira, C A; Martins, C M M R; Dos Santos, P P M; Balieiro, J C C; Rennó, F P; Fukushima, R S

    2018-04-18

    Feed intake assessment is a valuable tool for herd management decisions. The use of markers, either internal or external, is currently the most used technique for estimating feed intake in production animals. The experiment used 10 multiparous Holstein cows fed a corn silage-based diet, with 55:45 forage-to-concentrate ratio, the average fecal recovery (FR) of TiO 2 was higher than FR of Cr 2 O 3 , and both FR were more than unity. With internal markers, acetyl bromide lignin and cutin FR were lower than unity, and average FR for indigestible neutral detergent fiber (iNDF) and indigestible acid detergent fiber (iADF) was 1.5. The FR was unaffected by the fecal sampling procedure and appears to be an intrinsic property of each molecule and how it interacts with digesta. Of the 2 external markers, only Cr 2 O 3 produced accurate fecal output (FO) estimates and the same happened to dry matter digestibility (DMD) when iNDF and iADF were used. Estimates for DMD and FO were affected by sampling procedure; 72-h bulk [sub-sample from total feces collection (TFC)] sampling consistently produced accurate results. The grab (sub-samples taken at specific times during the day) sampling procedures were accurate when using either of the indigestible fibers (iNDF or iADF) to estimate DMD. However, grab sampling procedures can only be recommended when concomitant TFC is performed on at least one animal per treatment to determine FR. Under these conditions, Cr 2 O 3 is a suitable marker for estimating FO, and iNDF and iADF are adequate for estimating DMD. Moreover, the Cr 2 O 3 +iADF marker pair produces accurate dry matter intake estimates and deserves further attention in ruminant nutrition studies. The method of dosing the external markers is extremely important and greatly affects and determines results. Whichever the method, it must allow the animals to display normal feeding behavior and not affect performance. The grab sampling procedures can replace TFC (once FR is established), which may open new possibilities for pasture-based or collectively housed animals. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  6. Multicommuted flow system for the determination of glucose in animal blood serum exploiting enzymatic reaction and chemiluminescence detection

    PubMed Central

    Pires, Cherrine K.; Martelli, Patrícia B.; Lima, José L. F. C.; Saraiva, Maria Lúcia M. F. S.

    2003-01-01

    An automatic flow procedure based on multicommutation dedicated for the determination of glucose in animal blood serum using glucose oxidase with chemiluminescence detection is described. The flow manifold consisted of a set of three-way solenoid valves assembled to implement multicommutation. A microcomputer furnished with an electronic interface and software written in Quick BASIC 4.5 controlled the manifold and performed data acquisition. Glucose oxidase was immobilized on porous silica beads (glass aminopropyl) and packed in a minicolumn (15 × 5 mm). The procedure was based on the enzymatic degradation of glucose, producing hydrogen peroxide, which oxidized luminol in the presence of hexacyanoferrate(III), causing the chemiluminescence. The system was tested by analysing a set of serum animal samples without previous treatment. Results were in agreement with those obtained with the conventional method (LABTEST Kit) at the 95% confidence level. The detection limit and variation coefficient were estimated as 12.0 mg l−1 (99.7% confidence level) and 3.5% (n = 20), respectively. The sampling rate was about 60 determinations h−1 with sample concentrations ranging from 50 to 600 mg l−1 glucose. The consumptions of serum sample, hexacyanoferrate(III) and luminol were 46 μl, 10.0 mg and 0.2 mg/determination, respectively. PMID:18924619

  7. Childhood and Adolescent Animal Cruelty Methods and Their Possible Link to Adult Violent Crimes

    ERIC Educational Resources Information Center

    Hensley, Christopher; Tallichet, Suzanne E.

    2009-01-01

    Few researchers have investigated the potentially predictive power of childhood and adolescent animal cruelty methods as they are associated with subsequent interpersonal violence in adulthood. Based on a sample of 261 inmates at medium- and maximum-security prisons in a southern state, the present study examines the relationship between several…

  8. Examining the Relationship between Childhood Animal Cruelty Motives and Recurrent Adult Violent Crimes toward Humans

    ERIC Educational Resources Information Center

    Overton, Joshua C.; Hensley, Christopher; Tallichet, Suzanne E.

    2012-01-01

    Few researchers have studied the predictive ability of childhood animal cruelty motives as they are associated with later recurrent violence toward humans. Based on a sample of 180 inmates at one medium- and one maximum-security prison in a Southern state, the present study examines the relationship among several retrospectively identified motives…

  9. Technical note: Characterization of lipid constitution in Fourier transform infrared spectra and spectroscopic discrimination of animal-derived feedstuffs from different species.

    PubMed

    Gao, F; Han, L; Yang, Z; Xu, L; Liu, X

    2017-06-01

    The objective of the current work was to assess the capability of Fourier transform infrared (FT-IR) spectroscopy in combination with chemometric methods to discriminate animal-derived feedstuffs from different origins based on the lipid characteristics. A total of 82 lipid samples extracted from animal-derived feedstuffs, comprising porcine, poultry, bovine, ovine, and fish samples, were investigated by gas chromatography and FT-IR. The relationship between the lipid constitutions and the responding FT-IR spectral characteristics were explored. Results indicated that high correlations ( > 0.900) were found between the contents of MUFA and PUFA and FT-IR spectral data. In addition, the peak intensity at about 1,116 and 1,098 cm-1 showed a significant difference ( < 0.05) between ruminant and nonruminant animals; the change of peak ratio (1,116:1,098) was proved consistent with the degree of unsaturation of lipid from different animal species. Successful discrimination was further achieved among porcine, poultry, bovine, and ovine meat and bone meal (MBM) and fishmeal based on lipid characteristics by applying the FT-IR spectra coupled with chemometrics, for which the values of sensitivity and specificity were close to 1 and classification error were almost equal to 0.

  10. Modeling unobserved sources of heterogeneity in animal abundance using a Dirichlet process prior

    USGS Publications Warehouse

    Dorazio, R.M.; Mukherjee, B.; Zhang, L.; Ghosh, M.; Jelks, H.L.; Jordan, F.

    2008-01-01

    In surveys of natural populations of animals, a sampling protocol is often spatially replicated to collect a representative sample of the population. In these surveys, differences in abundance of animals among sample locations may induce spatial heterogeneity in the counts associated with a particular sampling protocol. For some species, the sources of heterogeneity in abundance may be unknown or unmeasurable, leading one to specify the variation in abundance among sample locations stochastically. However, choosing a parametric model for the distribution of unmeasured heterogeneity is potentially subject to error and can have profound effects on predictions of abundance at unsampled locations. In this article, we develop an alternative approach wherein a Dirichlet process prior is assumed for the distribution of latent abundances. This approach allows for uncertainty in model specification and for natural clustering in the distribution of abundances in a data-adaptive way. We apply this approach in an analysis of counts based on removal samples of an endangered fish species, the Okaloosa darter. Results of our data analysis and simulation studies suggest that our implementation of the Dirichlet process prior has several attractive features not shared by conventional, fully parametric alternatives. ?? 2008, The International Biometric Society.

  11. Initial animal studies of a wireless, batteryless, MEMS implant for cardiovascular applications.

    PubMed

    Najafi, Nader; Ludomirsky, Achiau

    2004-03-01

    This paper reports the results of the initial animal studies of a wireless, batteryless, implantable pressure sensor using microelectromechanical systems (MEMS) technology. The animal studies were acute and proved the functional feasibility of using MEMS technology for wireless bio sensing. The results are very encouraging and surpassed the majority of the application's requirements, including high sampling speed and high resolution. Based on the lessons learned, second generation wireless sensors are being developed that will provide total system solution.

  12. 9 CFR 113.3 - Sampling of biological products.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Sampling of biological products. 113.3 Section 113.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF... Applicability § 113.3 Sampling of biological products. Each licensee and permittee shall furnish representative...

  13. 9 CFR 113.3 - Sampling of biological products.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Sampling of biological products. 113.3 Section 113.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF... Applicability § 113.3 Sampling of biological products. Each licensee and permittee shall furnish representative...

  14. The Quality of Reporting of Measures of Precision in Animal Experiments in Implant Dentistry: A Methodological Study.

    PubMed

    Faggion, Clovis Mariano; Aranda, Luisiana; Diaz, Karla Tatiana; Shih, Ming-Chieh; Tu, Yu-Kang; Alarcón, Marco Antonio

    2016-01-01

    Information on precision of treatment-effect estimates is pivotal for understanding research findings. In animal experiments, which provide important information for supporting clinical trials in implant dentistry, inaccurate information may lead to biased clinical trials. The aim of this methodological study was to determine whether sample size calculation, standard errors, and confidence intervals for treatment-effect estimates are reported accurately in publications describing animal experiments in implant dentistry. MEDLINE (via PubMed), Scopus, and SciELO databases were searched to identify reports involving animal experiments with dental implants published from September 2010 to March 2015. Data from publications were extracted into a standardized form with nine items related to precision of treatment estimates and experiment characteristics. Data selection and extraction were performed independently and in duplicate, with disagreements resolved by discussion-based consensus. The chi-square and Fisher exact tests were used to assess differences in reporting according to study sponsorship type and impact factor of the journal of publication. The sample comprised reports of 161 animal experiments. Sample size calculation was reported in five (2%) publications. P values and confidence intervals were reported in 152 (94%) and 13 (8%) of these publications, respectively. Standard errors were reported in 19 (12%) publications. Confidence intervals were better reported in publications describing industry-supported animal experiments (P = .03) and with a higher impact factor (P = .02). Information on precision of estimates is rarely reported in publications describing animal experiments in implant dentistry. This lack of information makes it difficult to evaluate whether the translation of animal research findings to clinical trials is adequate.

  15. Comparison of two sampling and culture systems for detection of Salmonella enterica in the environment of a large animal hospital.

    PubMed

    Ruple-Czerniak, A; Bolte, D S; Burgess, B A; Morley, P S

    2014-07-01

    Nosocomial salmonellosis is an important problem in veterinary hospitals that treat horses and other large animals. Detection and mitigation of outbreaks and prevention of healthcare-associated infections often require detection of Salmonella enterica in the hospital environment. To compare 2 previously published methods for detecting environmental contamination with S. enterica in a large animal veterinary teaching hospital. Hospital-based comparison of environmental sampling techniques. A total of 100 pairs of environmental samples were collected from stalls used to house large animal cases (horses, cows or New World camelids) that were confirmed to be shedding S. enterica by faecal culture. Stalls were cleaned and disinfected prior to sampling, and the same areas within each stall were sampled for the paired samples. One method of detection used sterile, premoistened sponges that were cultured using thioglycolate enrichment before plating on XLT-4 agar. The other method used electrostatic wipes that were cultured using buffered peptone water, tetrathionate and Rappaport-Vassiliadis R10 broths before plating on XLT-4 agar. Salmonella enterica was recovered from 14% of samples processed using the electrostatic wipe sampling and culture procedure, whereas S. enterica was recovered from only 4% of samples processed using the sponge sampling and culture procedure. There was test agreement for 85 pairs of culture-negative samples and 3 pairs of culture-positive samples. However, the remaining 12 pairs of samples with discordant results created significant disagreement between the 2 detection methods (P<0.01). Persistence of Salmonella in the environment of veterinary hospitals can occur even with rigorous cleaning and disinfection. Use of sensitive methods for detection of environmental contamination is critical when detecting and mitigating this problem in veterinary hospitals. These results suggest that the electrostatic wipe sampling and culture method was more sensitive than the sponge sampling and culture method. © 2013 EVJ Ltd.

  16. The Impact of Animated Books on the Vocabulary and Language Development of Preschool-Aged Children in Two School Settings

    ERIC Educational Resources Information Center

    Broemmel, Amy D.; Moran, Mary Jane; Wooten, Deborah A.

    2015-01-01

    With the emergence of electronic media over the past two decades, young children have been found to have increased exposure to video games, computer-based activities, and electronic books (e-books). This study explores how exposure to animated ebooks impacts young children's literacy development. A stratified convenience sample (n = 24) was…

  17. Characterization of Methicillin Resistant Staphylococcus aureus isolated from human and animal samples in Egypt.

    PubMed

    Bendary, M M; Solyman, S M; Azab, M M; Mahmoud, N F; Hanora, A M

    2016-02-29

    Staphylococcus aureus (S. aureus) has been one of the most problematic pathogens. Methicillin Resistant S. aureus (MRSA) has emerged as a major concern for both human and animal. Antibiotic resistance genes dissemination might be possible between human and animal bacteria. The aim of this study is to show phenotypic and genotypic diversity of human and animal MRSA isolates. Antibiogram typing and biofilm production were used as a primary phenotypic typing tool for the characterization of (40) animal and (38) human MRSA isolates. Genetic typing based on sequencing of 16S rRNA gene and virulence gene profiles were done. Antimicrobial resistance profiles of the animal isolates showed little evidence of widespread of resistance, although this was seen in many human isolates. The biofilm production was detected in higher percentage among animal isolates. Based on the genetic typing and multiple antibiotic resistance (MAR) index, the majority of animal isolates clustered into lineages that were not found in human isolates. Animal and human MRSA isolates showed diversity in antibiotic resistance and virulence gene profiles may be due to host adaptation or chances for contamination between the two hosts were not present in our study.

  18. Sensitive Quantification of Aflatoxin B1 in Animal Feeds, Corn Feed Grain, and Yellow Corn Meal Using Immunomagnetic Bead-Based Recovery and Real-Time Immunoquantitative-PCR

    PubMed Central

    Babu, Dinesh; Muriana, Peter M.

    2014-01-01

    Aflatoxins are considered unavoidable natural mycotoxins encountered in foods, animal feeds, and feed grains. In this study, we demonstrate the application of our recently developed real-time immunoquantitative PCR (RT iq-PCR) assay for sensitive detection and quantification of aflatoxins in poultry feed, two types of dairy feed (1 and 2), horse feed, whole kernel corn feed grains, and retail yellow ground corn meal. Upon testing methanol/water (60:40) extractions of the above samples using competitive direct enzyme linked immunosorbent assay, the aflatoxin content was found to be <20 μg/kg. The RT iq-PCR assay exhibited high antigen hook effect in samples containing aflatoxin levels higher than the quantification limits (0.1–10 μg/kg), addressed by comparing the quantification results of undiluted and diluted extracts. In testing the reliability of the immuno-PCR assay, samples were spiked with 200 μg/kg of aflatoxin B1, but the recovery of spiked aflatoxin was found to be poor. Considering the significance of determining trace levels of aflatoxins and their serious implications for animal and human health, the RT iq-PCR method described in this study can be useful for quantifying low natural aflatoxin levels in complex matrices of food or animal feed samples without the requirement of extra sample cleanup. PMID:25474493

  19. Sensitive quantification of aflatoxin B1 in animal feeds, corn feed grain, and yellow corn meal using immunomagnetic bead-based recovery and real-time immunoquantitative-PCR.

    PubMed

    Babu, Dinesh; Muriana, Peter M

    2014-12-02

    Aflatoxins are considered unavoidable natural mycotoxins encountered in foods, animal feeds, and feed grains. In this study, we demonstrate the application of our recently developed real-time immunoquantitative PCR (RT iq-PCR) assay for sensitive detection and quantification of aflatoxins in poultry feed, two types of dairy feed (1 and 2), horse feed, whole kernel corn feed grains, and retail yellow ground corn meal. Upon testing methanol/water (60:40) extractions of the above samples using competitive direct enzyme linked immunosorbent assay, the aflatoxin content was found to be <20 μg/kg. The RT iq-PCR assay exhibited high antigen hook effect in samples containing aflatoxin levels higher than the quantification limits (0.1-10 μg/kg), addressed by comparing the quantification results of undiluted and diluted extracts. In testing the reliability of the immuno-PCR assay, samples were spiked with 200 μg/kg of aflatoxin B1, but the recovery of spiked aflatoxin was found to be poor. Considering the significance of determining trace levels of aflatoxins and their serious implications for animal and human health, the RT iq-PCR method described in this study can be useful for quantifying low natural aflatoxin levels in complex matrices of food or animal feed samples without the requirement of extra sample cleanup.

  20. Characterizing the rapid spread of porcine epidemic diarrhea virus (PEDV) through an animal food manufacturing facility.

    PubMed

    Schumacher, Loni L; Huss, Anne R; Cochrane, Roger A; Stark, Charles R; Woodworth, Jason C; Bai, Jianfa; Poulsen, Elizabeth G; Chen, Qi; Main, Rodger G; Zhang, Jianqiang; Gauger, Phillip C; Ramirez, Alejandro; Derscheid, Rachel J; Magstadt, Drew M; Dritz, Steve S; Jones, Cassandra K

    2017-01-01

    New regulatory and consumer demands highlight the importance of animal feed as a part of our national food safety system. Porcine epidemic diarrhea virus (PEDV) is the first viral pathogen confirmed to be widely transmissible in animal food. Because the potential for viral contamination in animal food is not well characterized, the objectives of this study were to 1) observe the magnitude of virus contamination in an animal food manufacturing facility, and 2) investigate a proposed method, feed sequencing, to decrease virus decontamination on animal food-contact surfaces. A U.S. virulent PEDV isolate was used to inoculate 50 kg swine feed, which was mixed, conveyed, and discharged into bags using pilot-scale feed manufacturing equipment. Surfaces were swabbed and analyzed for the presence of PEDV RNA by quantitative real-time polymerase chain reaction (qPCR). Environmental swabs indicated complete contamination of animal food-contact surfaces (0/40 vs. 48/48, positive baseline samples/total baseline samples, positive subsequent samples/total subsequent samples, respectively; P < 0.05) and near complete contamination of non-animal food-contact surfaces (0/24 vs. 16/18, positive baseline samples/total baseline samples, positive subsequent samples/total subsequent samples, respectively; P < 0.05). Flushing animal food-contact surfaces with low-risk feed is commonly used to reduce cross-contamination in animal feed manufacturing. Thus, four subsequent 50 kg batches of virus-free swine feed were manufactured using the same system to test its impact on decontaminating animal food-contact surfaces. Even after 4 subsequent sequences, animal food-contact surfaces retained viral RNA (28/33 positive samples/total samples), with conveying system being more contaminated than the mixer. A bioassay to test infectivity of dust from animal food-contact surfaces failed to produce infectivity. This study demonstrates the potential widespread viral contamination of surfaces in an animal food manufacturing facility and the difficulty of removing contamination using conventional feed sequencing, which underscores the importance for preventing viruses from entering and contaminating such facilities.

  1. Modeling abundance effects in distance sampling

    USGS Publications Warehouse

    Royle, J. Andrew; Dawson, D.K.; Bates, S.

    2004-01-01

    Distance-sampling methods are commonly used in studies of animal populations to estimate population density. A common objective of such studies is to evaluate the relationship between abundance or density and covariates that describe animal habitat or other environmental influences. However, little attention has been focused on methods of modeling abundance covariate effects in conventional distance-sampling models. In this paper we propose a distance-sampling model that accommodates covariate effects on abundance. The model is based on specification of the distance-sampling likelihood at the level of the sample unit in terms of local abundance (for each sampling unit). This model is augmented with a Poisson regression model for local abundance that is parameterized in terms of available covariates. Maximum-likelihood estimation of detection and density parameters is based on the integrated likelihood, wherein local abundance is removed from the likelihood by integration. We provide an example using avian point-transect data of Ovenbirds (Seiurus aurocapillus) collected using a distance-sampling protocol and two measures of habitat structure (understory cover and basal area of overstory trees). The model yields a sensible description (positive effect of understory cover, negative effect on basal area) of the relationship between habitat and Ovenbird density that can be used to evaluate the effects of habitat management on Ovenbird populations.

  2. Register-based predictors of violations of animal welfare legislation in dairy herds.

    PubMed

    Otten, N D; Nielsen, L R; Thomsen, P T; Houe, H

    2014-12-01

    The assessment of animal welfare can include resource-based or animal-based measures. Official animal welfare inspections in Denmark primarily control compliance with animal welfare legislation based on resource measures (e.g. housing system) and usually do not regard animal response parameters (e.g. clinical and behavioural observations). Herds selected for welfare inspections are sampled by a risk-based strategy based on existing register data. The aim of the present study was to evaluate register data variables as predictors of dairy herds with violations of the animal welfare legislation (VoAWL) defined as occurrence of at least one of the two most frequently violated measures found at recent inspections in Denmark, namely (a) presence of injured animals not separated from the rest of the group and/or (b) animals in a condition warranting euthanasia still being present in the herd. A total of 25 variables were extracted from the Danish Cattle Database and assessed as predictors using a multivariable logistic analysis of a data set including 73 Danish dairy herds, which all had more than 100 cows and cubicle loose-housing systems. Univariable screening was used to identify variables associated with VoAWL at a P-value<0.2 for the inclusion in a multivariable logistic regression analysis. Backward selection procedures identified the following variables for the final model predictive of VoAWL: increasing standard deviation of milk yield for first lactation cows, high bulk tank somatic cell count (⩾250 000 cells/ml) and suspiciously low number of recorded veterinary treatments (⩽25 treatments/100 cow years). The identified predictors may be explained by underlying management factors leading to impaired animal welfare in the herd, such as poor hygiene, feeding and management of dry or calving cows and sick animals. However, further investigations are required for causal inferences to be established.

  3. Prevalence of Escherichia coli O157:H7 and performance by beef feedlot cattle given Lactobacillus direct-fed microbials.

    PubMed

    Brashears, M M; Galyean, M L; Loneragan, G H; Mann, J E; Killinger-Mann, K

    2003-05-01

    Fecal shedding of Escherichia coli O157:H7, the prevalence of Escherichia coli O157:H7 in pens and on carcasses and hides, and cattle performance as a result of daily dietary supplementation with Lactobacillus-based direct-fed microbials (DFMs) were evaluated in a feeding trial involving 180 beef steers. Steers were evaluated for shedding of E. coli O157:H7 by an immunomagnetic separation technique on arrival at the feedlot, just before treatment with the DFMs, and every 14 days thereafter until slaughter. Composite pen fecal samples were collected every 14 days (alternating weeks with animal testing), and prevalence on hides and carcasses at slaughter was also evaluated. Feedlot performance (body weight gain and feed intake) was measured for the period during which the DFMs were fed. Gain efficiency was calculated as the ratio of weight gain to feed intake. Lactobacillus acidophilus NPC 747 decreased (P < 0.01) the shedding of E. coli O157:H7 in the feces of individual cattle during the feeding period. E. coli O157:H7 was approximately twice as likely to be detected in control animal samples as in samples from animals receiving L. acidophilus NPC 747. In addition, DFM supplementation decreased (P < 0.05) the number of E. coli O157:H7-positive hide samples at harvest and the number of pens testing positive for the pathogen. Body weight gains (on a live or carcass basis) and feed intakes during the DFM supplementation period did not differ among treatments. Gain efficiencies on a live-weight basis did not differ among treatments, but carcass-based gain/feed ratios tended (P < 0.06) to be better for animals receiving the two DFM treatments than for control animals. The results of this study suggest that the feeding of a Lactobacillus-based DFM to cattle will decrease, but not eliminate, fecal shedding of E. coli O157:H7, as well as contamination on hides, without detrimental effects on performance.

  4. Extraction of 3,4,4'-Trichlorocarbanilide from Rat Fecal Samples for Determination by High Pressure Liquid Chromatography-Tandem Mass Spectrometry.

    PubMed

    Kennedy, Rebekah C; Fling, Russell R; Terry, Paul D; Menn, Fu-Min; Chen, Jiangang; Borman, Christopher J

    2015-07-15

    Triclocarban (3,4,4'-Trichlorocarbanilide; TCC) in the environment has been well documented. Methods have been developed to monitor TCC levels from various matrices including water, sediment, biosolids, plants, blood and urine; however, no method has been developed to document the concentration of TCC in fecal content after oral exposure in animal studies. In the present study, we developed and validated a method that uses liquid extraction coupled with HPLC-MS/MS determination to measure TCC in feces. The limit of detection and limit of quantitation in control rats without TCC exposure was 69.0 ng/g and 92.9 ng/g of feces, respectively. The base levels of TCC in feces were lower than LOD. At 12 days of treatment, the fecal TCC concentration increased to 2220 µg/g among 0.2% w/w exposed animals. The concentration in fecal samples decreased over the washout period in 0.2% w/w treated animals to 0.399 µ/g feces after exposure was removed for 28 days. This method required a small amount of sample (0.1 g) with simple sample preparation. Given its sensitivity and efficiency, this method may be useful for monitoring TCC exposure in toxicological studies of animals.

  5. A survey of hemoparasites and ectoparasites in Nasua nasua Linnaeus, 1766 with a redescription of Hepatozoon procyonis Richards, 1961 based on morphological and molecular data.

    PubMed

    da Silva, Maria Regina Lucas; Fornazari, Felipe; Martins, Thiago Fernandes; Hippólito, Alícia Giolo; Rolim, Luna Scarpari; Bisca, Jacqueline Muniz; Teixeira, Carlos Roberto; O'Dwyer, Lucia Helena

    2018-07-01

    Hemoparasites are vector-borne parasites that infect wild carnivores worldwide. Since data on hemoparasite infections in Nasua nasua from Brazil are lacking, the aim of this study was to investigate the occurrence of hemoparasites and ectoparasites in N. nasua from different areas of Brazil. Blood samples and ectoparasites from 83 N. nasua were collected in Botucatu, Palmital, and São Paulo municipalities. Samples were screened via microscopy and molecular methods to detect hemoparasites. Tissues from two N. nasua were obtained for histopathological and molecular analyses. All 83 samples were negative for piroplasms on morphological and molecular examination. Thin blood smears of nine animals were positive for Hepatozoon gamonts. The gamonts shared morphological characteristics of Hepatozoon procyonis. Meronts were detected in the liver and spleen tissue of one animal. Twenty-one blood samples and four tissue samples were PCR positive for Hepatozoon sp. The sequences obtained were 97% identical to those of Hepatozoon felis, Hepatozoon ursi, and Hepatozoon sp. Based on searches for similarity and morphology, we identified the sequences as belonging to H. procyonis. This study provides epidemiological data on hemoparasite infections and redescribe H. procyonis based on morphological, morphometrical, and molecular analyses.

  6. An empirical comparison of isolate-based and sample-based definitions of antimicrobial resistance and their effect on estimates of prevalence.

    PubMed

    Humphry, R W; Evans, J; Webster, C; Tongue, S C; Innocent, G T; Gunn, G J

    2018-02-01

    Antimicrobial resistance is primarily a problem in human medicine but there are unquantified links of transmission in both directions between animal and human populations. Quantitative assessment of the costs and benefits of reduced antimicrobial usage in livestock requires robust quantification of transmission of resistance between animals, the environment and the human population. This in turn requires appropriate measurement of resistance. To tackle this we selected two different methods for determining whether a sample is resistant - one based on screening a sample, the other on testing individual isolates. Our overall objective was to explore the differences arising from choice of measurement. A literature search demonstrated the widespread use of testing of individual isolates. The first aim of this study was to compare, quantitatively, sample level and isolate level screening. Cattle or sheep faecal samples (n=41) submitted for routine parasitology were tested for antimicrobial resistance in two ways: (1) "streak" direct culture onto plates containing the antimicrobial of interest; (2) determination of minimum inhibitory concentration (MIC) of 8-10 isolates per sample compared to published MIC thresholds. Two antibiotics (ampicillin and nalidixic acid) were tested. With ampicillin, direct culture resulted in more than double the number of resistant samples than the MIC method based on eight individual isolates. The second aim of this study was to demonstrate the utility of the observed relationship between these two measures of antimicrobial resistance to re-estimate the prevalence of antimicrobial resistance from a previous study, in which we had used "streak" cultures. Boot-strap methods were used to estimate the proportion of samples that would have tested resistant in the historic study, had we used the isolate-based MIC method instead. Our boot-strap results indicate that our estimates of prevalence of antimicrobial resistance would have been considerably lower in the historic study had the MIC method been used. Finally we conclude that there is no single way of defining a sample as resistant to an antimicrobial agent. The method used greatly affects the estimated prevalence of antimicrobial resistance in a sampled population of animals, thus potentially resulting in misleading results. Comparing methods on the same samples allows us to re-estimate the prevalence from other studies, had other methods for determining resistance been used. The results of this study highlight the importance of establishing what the most appropriate measure of antimicrobial resistance is, for the proposed purpose of the results. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Detection of Francisella tularensis-Specific Antibodies in Patients with Tularemia by a Novel Competitive Enzyme-Linked Immunosorbent Assay

    PubMed Central

    Sharma, Neekun; Hotta, Akitoyo; Yamamoto, Yoshie; Fujita, Osamu; Uda, Akihiko; Morikawa, Shigeru; Yamada, Akio

    2013-01-01

    A novel competitive enzyme-linked immunosorbent assay (cELISA) was developed and evaluated for detection of antibodies against Francisella tularensis in humans. The assay is based on the ability of serum antibodies to inhibit the binding of monoclonal antibodies (MAbs) directed against F. tularensis lipopolysaccharide antigens. The assay was evaluated using serum samples of tularemia patients, inactivated F. tularensis-immunized rabbits, and F. tularensis-infected mice. Antibodies against F. tularensis were successfully detected in serum samples of tularemia patients as well as the immunized and infected animals. The cELISA method was compared to indirect ELISA (iELISA) and the commonly used microagglutination test (MA) using serum samples of 19 tularemia patients and 50 healthy individuals. The sensitivity and specificity of cELISA were 93.9 and 96.1%, respectively, in comparison to the iELISA. MA was less sensitive than cELISA with a sensitivity and specificity of only 81.8 and 98.0%, respectively. A high degree of correlation (R2 = 0.8226) was observed between cELISA and iELISA results. The novel cELISA developed in this study appears to be highly sensitive and specific for serodiagnosis of human tularemia. The potential of the MAb-based cELISA to be used in both human and animal samples emphasizes its usefulness for serological survey of tularemia among multiple animal species. PMID:23114700

  8. 9 CFR 147.8 - Procedures for preparing egg yolk samples for diagnostic tests.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Procedures for preparing egg yolk samples for diagnostic tests. 147.8 Section 147.8 Animals and Animal Products ANIMAL AND PLANT HEALTH... IMPROVEMENT PLAN Blood Testing Procedures § 147.8 Procedures for preparing egg yolk samples for diagnostic...

  9. 9 CFR 147.8 - Procedures for preparing egg yolk samples for diagnostic tests.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Procedures for preparing egg yolk samples for diagnostic tests. 147.8 Section 147.8 Animals and Animal Products ANIMAL AND PLANT HEALTH... IMPROVEMENT PLAN Blood Testing Procedures § 147.8 Procedures for preparing egg yolk samples for diagnostic...

  10. 9 CFR 147.8 - Procedures for preparing egg yolk samples for diagnostic tests.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Procedures for preparing egg yolk samples for diagnostic tests. 147.8 Section 147.8 Animals and Animal Products ANIMAL AND PLANT HEALTH... IMPROVEMENT PLAN Blood Testing Procedures § 147.8 Procedures for preparing egg yolk samples for diagnostic...

  11. 9 CFR 147.8 - Procedures for preparing egg yolk samples for diagnostic tests.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Procedures for preparing egg yolk samples for diagnostic tests. 147.8 Section 147.8 Animals and Animal Products ANIMAL AND PLANT HEALTH... IMPROVEMENT PLAN Blood Testing Procedures § 147.8 Procedures for preparing egg yolk samples for diagnostic...

  12. Evaluation of a monoclonal antibody-based rapid immunochromatographic test for direct detection of rabies virus in the brain of humans and animals.

    PubMed

    Ahmed, Kamruddin; Wimalaratne, Omala; Dahal, Narapati; Khawplod, Pakamatz; Nanayakkara, Susilakanthi; Rinzin, Karma; Perera, Devika; Karunanayake, Dushantha; Matsumoto, Takashi; Nishizono, Akira

    2012-04-01

    Rabies diagnosis uses a direct fluorescent antibody test (FAT) that is difficult, costly, and time-consuming, and requires trained personnel. We developed a rapid immunochromatographic test (RICT) for the diagnosis of rabies. The efficacy of the RICT was compared with that of the FAT. Brain samples were collected from humans, dogs, cats, and other animals in Sri Lanka (n = 248), Bhutan (n = 27), and Thailand (n = 228). The sensitivity (0.74-0.95), specificity (0.98-1.0), positive predictive value (0.98-1.0), negative predictive value (0.75-0.97), accuracy (0.91-0.98), and kappa measure of agreement (0.79-0.93) were all satisfactory for animal samples and samples preserved in 50% glycerol saline solution. Because the RICT showed high sensitivity but low specificity with human brain samples, it is unsuitable for confirming rabies in humans. No amino acid substitutions were found in the antibody attachment sites of the nucleoprotein gene with FAT-positive, RICT-negative samples. The RICT is reliable, user friendly, rapid, robust, and can be used in laboratories with a modest infrastructure.

  13. Population size and stopover duration estimation using mark–resight data and Bayesian analysis of a superpopulation model

    USGS Publications Warehouse

    Lyons, James E.; Kendall, William L.; Royle, J. Andrew; Converse, Sarah J.; Andres, Brad A.; Buchanan, Joseph B.

    2016-01-01

    We present a novel formulation of a mark–recapture–resight model that allows estimation of population size, stopover duration, and arrival and departure schedules at migration areas. Estimation is based on encounter histories of uniquely marked individuals and relative counts of marked and unmarked animals. We use a Bayesian analysis of a state–space formulation of the Jolly–Seber mark–recapture model, integrated with a binomial model for counts of unmarked animals, to derive estimates of population size and arrival and departure probabilities. We also provide a novel estimator for stopover duration that is derived from the latent state variable representing the interim between arrival and departure in the state–space model. We conduct a simulation study of field sampling protocols to understand the impact of superpopulation size, proportion marked, and number of animals sampled on bias and precision of estimates. Simulation results indicate that relative bias of estimates of the proportion of the population with marks was low for all sampling scenarios and never exceeded 2%. Our approach does not require enumeration of all unmarked animals detected or direct knowledge of the number of marked animals in the population at the time of the study. This provides flexibility and potential application in a variety of sampling situations (e.g., migratory birds, breeding seabirds, sea turtles, fish, pinnipeds, etc.). Application of the methods is demonstrated with data from a study of migratory sandpipers.

  14. Standard of reporting animal-based experimental research in Indian Journal of Pharmacology.

    PubMed

    Aiman, Umme; Rahman, Syed Ziaur

    2015-01-01

    The objective of present study was to survey and determine the reporting standards of animal studies published during three years from 2012 to 2014 in the Indian Journal of Pharmacology (IJP). All issues of IJP published in the year 2012, 2013 and 2014 were reviewed to identify animal studies. Each animal study was searched for 15 parameters specifically designed to review standards of animal experimentation and research methodology. All published studies had clearly defined aims and objectives while a statement on ethical clearance about the study protocol was provided in 97% of papers. Information about animal strain and sex was given in 91.8% and 90% of papers respectively. Age of experimental animals was mentioned by 44.4% papers while source of animals was given in 50.8% papers. Randomization was reported by 37.4% while 9.9% studies reported blinding. Only 3.5% studies mentioned any limitations of their work. Present study demonstrates relatively good reporting standards in animal studies published in IJP. The items which need to be improved are randomization, blinding, sample size calculation, stating the limitations of study, sources of support and conflict of interest. The knowledge shared in the present paper could be used for better reporting of animal based experiments.

  15. Development of an On-animal Separation-based Sensor for Monitoring Drug Metabolism in Freely Roaming Sheep

    PubMed Central

    Scott, David E.; Willis, Sean D.; Gabbert, Seth; Johnson, Dave A.; Naylor, Erik; Janle, Elsa M.; Krichevsky, Janice E.; Lunte, Craig E.; Lunte, Susan M.

    2015-01-01

    The development of an on-animal separation-based sensor that can be employed for monitoring drug metabolism in a freely roaming sheep is described. The system consists of microdialysis sampling coupled directly to microchip electrophoresis with electrochemical detection (MD-ME-EC). Separations were accomplished using an all-glass chip with integrated platinum working and reference electrodes. Discrete samples from the microdialysis flow were introduced into the electrophoresis chip using a flow-gated injection approach. Electrochemical detection was accomplished in-channel using a two-electrode isolated potentiostat. Nitrite was separated by microchip electrophoresis using reverse polarity and a run buffer consisting of 50 mM phosphate at pH 7.4. The entire system was under telemetry control. The system was first tested with rats to monitor the production of nitrite following introduction of nitroglycerin into the subdermal tissue using a linear probe. The data acquired using the on-line MD-ME-EC system was compared to that obtained off-line analysis by liquid chromatography with electrochemical detection (LC-EC), using a second microdialysis probe implanted parallel to the first probe in the same animal. The MD-ME-EC device was then used on-animal to monitor the subdermal metabolism of nitroglycerin in sheep. The ultimate goal is to use this device to simultaneously monitor drug metabolism and behavior in a freely roaming animal. PMID:25697221

  16. Serological evidence for hepatitis e virus infection in laboratory monkeys and pigs in animal facilities in Japan.

    PubMed

    Yamamoto, Hiroshi; Li, Tian-Cheng; Koshimoto, Chihiro; Ito, Kaori; Kita, Masakazu; Miyashita, Nobumoto; Arikawa, Jiro; Yagami, Kenichi; Asano, Masahide; Tezuka, Hideo; Suzuki, Noboru; Kurosawa, Tsutomu; Shibahara, Toshiyuki; Furuya, Masato; Mohri, Shirou; Sato, Hiroshi; Ohsawa, Kazutaka; Ibuki, Kentaro; Takeda, Naokazu

    2008-07-01

    In laboratory animal facilities, monkeys and pigs are used for animal experiments, but the details of hepatitis E virus (HEV) infection in these animals are unknown. The risk of infection from laboratory animals to humans has become a concern; therefore, much attention should be paid to the handling of these animals during their care and use, including surgical procedures performed on infected animals. In this connection, serum samples collected from 916 monkeys and 77 pigs kept in 23 animal facilities belonging to the Japanese Association of Laboratory Animal Facilities of National University Corporations (JALAN) and the Japanese Association of Laboratory Animal Facilities of Public and Private Universities (JALAP) in Japan were examined for the purpose of detecting antibodies to HEV and HEV RNA by using ELISA and RT-PCR, respectively. One hundred and seven serum samples of 916 (11.7%) monkeys were positive for anti-HEV IgG, and 7 and 17 serum samples of 916 (0.8% and 5.3%) monkeys were positive for anti-HEV IgM and IgA, respectively. Thirty-six samples from 62 (58.1%) farm pigs were positive for anti-HEV IgG, whereas all samples tested from miniature pigs were negative (0/15, 0%). Seven samples from 62 (9.1%) farm pigs and 7 samples from 916 (0.8%) monkeys were positive for IgM antibody, but these HEV-IgM antibody positive serum samples were HEV-RNA negative by RT-PCR. The IgM antibody positive rate (9.1%) of farm pigs was much higher than that of monkeys (0.8%). These results suggest the relative levels of risk of HEV infection from these animals to animal handlers and researchers who work with them in laboratory animal facilities.

  17. A dynamic Brownian bridge movement model to estimate utilization distributions for heterogeneous animal movement.

    PubMed

    Kranstauber, Bart; Kays, Roland; Lapoint, Scott D; Wikelski, Martin; Safi, Kamran

    2012-07-01

    1. The recently developed Brownian bridge movement model (BBMM) has advantages over traditional methods because it quantifies the utilization distribution of an animal based on its movement path rather than individual points and accounts for temporal autocorrelation and high data volumes. However, the BBMM assumes unrealistic homogeneous movement behaviour across all data. 2. Accurate quantification of the utilization distribution is important for identifying the way animals use the landscape. 3. We improve the BBMM by allowing for changes in behaviour, using likelihood statistics to determine change points along the animal's movement path. 4. This novel extension, outperforms the current BBMM as indicated by simulations and examples of a territorial mammal and a migratory bird. The unique ability of our model to work with tracks that are not sampled regularly is especially important for GPS tags that have frequent failed fixes or dynamic sampling schedules. Moreover, our model extension provides a useful one-dimensional measure of behavioural change along animal tracks. 5. This new method provides a more accurate utilization distribution that better describes the space use of realistic, behaviourally heterogeneous tracks. © 2012 The Authors. Journal of Animal Ecology © 2012 British Ecological Society.

  18. Recovery and Enumeration of Cryptosporidium parvum from Animal Fecal Matrices

    PubMed Central

    Davies, Cheryl M.; Kaucner, Christine; Deere, Daniel; Ashbolt, Nicholas J.

    2003-01-01

    Accurate quantification of Cryptosporidium parvum oocysts in animal fecal deposits on land is an essential starting point for estimating watershed C. parvum loads. Due to the general poor performance and variable recovery efficiency of existing enumeration methods, protocols were devised based on initial dispersion of oocysts from feces by vortexing in 2 mM tetrasodium pyrophosphate, followed by immunomagnetic separation. The protocols were validated by using an internal control seed preparation to determine the levels of oocyst recovery for a range of fecal types. The levels of recovery of 102 oocysts from cattle feces (0.5 g of processed feces) ranged from 31 to 46%, and the levels of recovery from sheep feces (0.25 g of processed feces) ranged from 21% to 35%. The within-sample coefficients of variation for the percentages of recovery from five replicates ranged from 10 to 50%. The ranges for levels of recovery of oocysts from cattle, kangaroo, pig, and sheep feces (juveniles and adults) collected in a subsequent watershed animal fecal survey were far wider than the ranges predicted by the validation data. Based on the use of an internal control added to each fecal sample, the levels of recovery ranged from 0 to 83% for cattle, from 4 to 62% for sheep, from 1 to 42% for pigs, and from 40 to 73% for kangaroos. Given the variation in the levels of recovery of oocysts from different fecal matrices, it is recommended that an internal control be added to at least one replicate of every fecal sample analyzed to determine the percentage of recovery. Depending on the animal type and based on the lowest approximate percentages of recovery, between 10 and 100 oocysts g of feces−1 must be present to be detected. PMID:12732556

  19. PCR detection and quantitation of predominant anaerobic bacteria in human and animal fecal samples.

    PubMed Central

    Wang, R F; Cao, W W; Cerniglia, C E

    1996-01-01

    PCR procedures based on 16S rRNA gene sequences specific for 12 anaerobic bacteria that predominate in the human intestinal tract were developed and used for quantitative detection of these species in human (adult and baby) feces and animal (rat, mouse, cat, dog, monkey, and rabbit) feces. Fusobacterium prausnitzii, Peptostreptococcus productus, and Clostridium clostridiiforme had high PCR titers (the maximum dilutions for positive PCR results ranged from 10(-3) to 10(-8)) in all of the human and animal fecal samples tested. Bacteroides thetaiotaomicron, Bacteroides vulgatus, and Eubacterium limosum also showed higher PCR titers (10(-2) to 10(-6)) in adult human feces. The other bacteria tested, including Escherichia coli, Bifidobacterium adolescentis, Bifidobacterium longum, Lactobacillus acidophilus, Eubacterium biforme, and Bacteroides distasonis, were either at low PCR titers (less than 10(-2)) or not detected by PCR. The reported PCR procedure including the fecal sample preparation method is simplified and rapid and eliminates the DNA isolation steps. PMID:8919784

  20. 9 CFR 381.146 - Sampling at official establishments.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Sampling at official establishments. 381.146 Section 381.146 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF... Establishments; Processing Inspection and Other Reinspections; Processing Requirements § 381.146 Sampling at...

  1. 9 CFR 381.146 - Sampling at official establishments.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Sampling at official establishments. 381.146 Section 381.146 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF... Establishments; Processing Inspection and Other Reinspections; Processing Requirements § 381.146 Sampling at...

  2. A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas)

    PubMed Central

    Wang, Jiann-Hsiung; Chou, Shih-Jen; Li, Tsung-Hsien; Leu, Ming-Yih; Ho, Hsiao-Kuan

    2017-01-01

    Cytokines are fundamental for a functioning immune system, and thus potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a probe-based quantitative gene expression assay for immunological assessment of captive beluga whales (Delphinapterus leucas) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater™ solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset. PMID:28970970

  3. A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas).

    PubMed

    Tsai, Ming-An; Chen, I-Hua; Wang, Jiann-Hsiung; Chou, Shih-Jen; Li, Tsung-Hsien; Leu, Ming-Yih; Ho, Hsiao-Kuan; Yang, Wei Cheng

    2017-01-01

    Cytokines are fundamental for a functioning immune system, and thus potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a probe-based quantitative gene expression assay for immunological assessment of captive beluga whales ( Delphinapterus leucas ) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNA later ™ solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset.

  4. Molecular characterization of bacteriophages for microbial source tracking in Korea.

    PubMed

    Lee, Jung Eun; Lim, Mi Young; Kim, Sei Yoon; Lee, Sunghee; Lee, Heetae; Oh, Hyun-Myung; Hur, Hor-Gil; Ko, Gwangpyo

    2009-11-01

    We investigated coliphages from various fecal sources, including humans and animals, for microbial source tracking in South Korea. Both somatic and F+-specific coliphages were isolated from 43 fecal samples from farms, wild animal habitats, and human wastewater plants. Somatic coliphages were more prevalent and abundant than F+ coliphages in all of the tested fecal samples. We further characterized 311 F+ coliphage isolates using RNase sensitivity assays, PCR and reverse transcription-PCR, and nucleic acid sequencing. Phylogenetic analyses were performed based on the partial nucleic acid sequences of 311 F+ coliphages from various sources. F+ RNA coliphages were most prevalent among geese (95%) and were least prevalent in cows (5%). Among the genogroups of F+ RNA coliphages, most F+ coliphages isolated from animal fecal sources belonged to either group I or group IV, and most from human wastewater sources were in group II or III. Some of the group I coliphages were present in both human and animal source samples. F+ RNA coliphages isolated from various sources were divided into two main clusters. All F+ RNA coliphages isolated from human wastewater were grouped with Qbeta-like phages, while phages isolated from most animal sources were grouped with MS2-like phages. UniFrac significance statistical analyses revealed significant differences between human and animal bacteriophages. In the principal coordinate analysis (PCoA), F+ RNA coliphages isolated from human waste were distinctively separate from those isolated from other animal sources. However, F+ DNA coliphages were not significantly different or separate in the PCoA. These results demonstrate that proper analysis of F+ RNA coliphages can effectively distinguish fecal sources.

  5. Development and validation of a real-time PCR assay for the detection of Toxoplasma gondii DNA in animal and meat samples.

    PubMed

    Marino, Anna Maria Fausta; Percipalle, Maurizio; Giunta, Renato Paolo; Salvaggio, Antonio; Caracappa, Giulia; Alfonzetti, Tiziana; Aparo, Alessandra; Reale, Stefano

    2017-03-01

    We report a rapid and reliable method for the detection of Toxoplasma gondii in meat and animal tissues based on real-time polymerase chain reaction (PCR). Samples were collected from cattle, small ruminants, horses, and pigs raised or imported into Sicily, Italy. All DNA preparations were assayed by real-time PCR tests targeted to a 98-bp long fragment in the AF 529-bp repeat element and to the B1 gene using specific primers. Diagnostic sensitivity (100%), diagnostic specificity (100%), limit of detection (0.01 pg), efficiency (92-109%), and precision (mean coefficient of variation = 0.60%), repeatability (100%), reproducibility (100%), and robustness were evaluated using 240 DNA extracted samples (120 positives and 120 negative as per the OIE nested PCR method) from different matrices. Positive results were confirmed by the repetition of both real-time and nested PCR assays. Our study demonstrates the viability of a reliable, rapid, and specific real-time PCR on a large scale to monitor contamination with Toxoplasma cysts in meat and animal specimens. This validated method can be used for postmortem detection in domestic and wild animals and for food safety purposes.

  6. Characterizing the rapid spread of porcine epidemic diarrhea virus (PEDV) through an animal food manufacturing facility

    PubMed Central

    Schumacher, Loni L.; Huss, Anne R.; Cochrane, Roger A.; Stark, Charles R.; Woodworth, Jason C.; Bai, Jianfa; Poulsen, Elizabeth G.; Chen, Qi; Main, Rodger G.; Zhang, Jianqiang; Gauger, Phillip C.; Ramirez, Alejandro; Derscheid, Rachel J.; Magstadt, Drew M.; Dritz, Steve S.

    2017-01-01

    New regulatory and consumer demands highlight the importance of animal feed as a part of our national food safety system. Porcine epidemic diarrhea virus (PEDV) is the first viral pathogen confirmed to be widely transmissible in animal food. Because the potential for viral contamination in animal food is not well characterized, the objectives of this study were to 1) observe the magnitude of virus contamination in an animal food manufacturing facility, and 2) investigate a proposed method, feed sequencing, to decrease virus decontamination on animal food-contact surfaces. A U.S. virulent PEDV isolate was used to inoculate 50 kg swine feed, which was mixed, conveyed, and discharged into bags using pilot-scale feed manufacturing equipment. Surfaces were swabbed and analyzed for the presence of PEDV RNA by quantitative real-time polymerase chain reaction (qPCR). Environmental swabs indicated complete contamination of animal food-contact surfaces (0/40 vs. 48/48, positive baseline samples/total baseline samples, positive subsequent samples/total subsequent samples, respectively; P < 0.05) and near complete contamination of non-animal food-contact surfaces (0/24 vs. 16/18, positive baseline samples/total baseline samples, positive subsequent samples/total subsequent samples, respectively; P < 0.05). Flushing animal food-contact surfaces with low-risk feed is commonly used to reduce cross-contamination in animal feed manufacturing. Thus, four subsequent 50 kg batches of virus-free swine feed were manufactured using the same system to test its impact on decontaminating animal food-contact surfaces. Even after 4 subsequent sequences, animal food-contact surfaces retained viral RNA (28/33 positive samples/total samples), with conveying system being more contaminated than the mixer. A bioassay to test infectivity of dust from animal food-contact surfaces failed to produce infectivity. This study demonstrates the potential widespread viral contamination of surfaces in an animal food manufacturing facility and the difficulty of removing contamination using conventional feed sequencing, which underscores the importance for preventing viruses from entering and contaminating such facilities. PMID:29095859

  7. Feasibility Study of NMR Based Serum Metabolomic Profiling to Animal Health Monitoring: A Case Study on Iron Storage Disease in Captive Sumatran Rhinoceros (Dicerorhinus sumatrensis).

    PubMed

    Watanabe, Miki; Roth, Terri L; Bauer, Stuart J; Lane, Adam; Romick-Rosendale, Lindsey E

    2016-01-01

    A variety of wildlife species maintained in captivity are susceptible to iron storage disease (ISD), or hemochromatosis, a disease resulting from the deposition of excess iron into insoluble iron clusters in soft tissue. Sumatran rhinoceros (Dicerorhinus sumatrensis) is one of the rhinoceros species that has evolutionarily adapted to a low-iron diet and is susceptible to iron overload. Hemosiderosis is reported at necropsy in many African black and Sumatran rhinoceroses but only a small number of animals reportedly die from hemochromatosis. The underlying cause and reasons for differences in susceptibility to hemochromatosis within the taxon remains unclear. Although serum ferritin concentrations have been useful in monitoring the progression of ISD in many species, there is some question regarding their value in diagnosing hemochromatosis in the Sumatran rhino. To investigate the metabolic changes during the development of hemochromatosis and possibly increase our understanding of its progression and individual susceptibility differences, the serum metabolome from a Sumatran rhinoceros was investigated by nuclear magnetic resonance (NMR)-based metabolomics. The study involved samples from female rhinoceros at the Cincinnati Zoo (n = 3), including two animals that died from liver failure caused by ISD, and the Sungai Dusun Rhinoceros Conservation Centre in Peninsular Malaysia (n = 4). Principal component analysis was performed to visually and statistically compare the metabolic profiles of the healthy animals. The results indicated that significant differences were present between the animals at the zoo and the animals in the conservation center. A comparison of the 43 serum metabolomes of three zoo rhinoceros showed two distinct groupings, healthy (n = 30) and unhealthy (n = 13). A total of eighteen altered metabolites were identified in healthy versus unhealthy samples. Results strongly suggest that NMR-based metabolomics is a valuable tool for animal health monitoring and may provide insight into the progression of this and other insidious diseases.

  8. 40 CFR 79.67 - Glial fibrillary acidic protein assay.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... immunoreactivity of individual samples (both control and exposed groups) with that of the sample used to generate... the control groups is normalized to 100 percent and all data are expressed as a percentage of control... emission-exposed and unexposed control animals. It is based on modifications (O'Callaghan & Miller 1985 in...

  9. 40 CFR 79.67 - Glial fibrillary acidic protein assay.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... immunoreactivity of individual samples (both control and exposed groups) with that of the sample used to generate... the control groups is normalized to 100 percent and all data are expressed as a percentage of control... emission-exposed and unexposed control animals. It is based on modifications (O'Callaghan & Miller 1985 in...

  10. 40 CFR 79.67 - Glial fibrillary acidic protein assay.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... immunoreactivity of individual samples (both control and exposed groups) with that of the sample used to generate... the control groups is normalized to 100 percent and all data are expressed as a percentage of control... emission-exposed and unexposed control animals. It is based on modifications (O'Callaghan & Miller 1985 in...

  11. 40 CFR 79.67 - Glial fibrillary acidic protein assay.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... immunoreactivity of individual samples (both control and exposed groups) with that of the sample used to generate... the control groups is normalized to 100 percent and all data are expressed as a percentage of control... emission-exposed and unexposed control animals. It is based on modifications (O'Callaghan & Miller 1985 in...

  12. 40 CFR 79.67 - Glial fibrillary acidic protein assay.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... immunoreactivity of individual samples (both control and exposed groups) with that of the sample used to generate... the control groups is normalized to 100 percent and all data are expressed as a percentage of control... emission-exposed and unexposed control animals. It is based on modifications (O'Callaghan & Miller 1985 in...

  13. On marker-based parentage verification via non-linear optimization.

    PubMed

    Boerner, Vinzent

    2017-06-15

    Parentage verification by molecular markers is mainly based on short tandem repeat markers. Single nucleotide polymorphisms (SNPs) as bi-allelic markers have become the markers of choice for genotyping projects. Thus, the subsequent step is to use SNP genotypes for parentage verification as well. Recent developments of algorithms such as evaluating opposing homozygous SNP genotypes have drawbacks, for example the inability of rejecting all animals of a sample of potential parents. This paper describes an algorithm for parentage verification by constrained regression which overcomes the latter limitation and proves to be very fast and accurate even when the number of SNPs is as low as 50. The algorithm was tested on a sample of 14,816 animals with 50, 100 and 500 SNP genotypes randomly selected from 40k genotypes. The samples of putative parents of these animals contained either five random animals, or four random animals and the true sire. Parentage assignment was performed by ranking of regression coefficients, or by setting a minimum threshold for regression coefficients. The assignment quality was evaluated by the power of assignment (P[Formula: see text]) and the power of exclusion (P[Formula: see text]). If the sample of putative parents contained the true sire and parentage was assigned by coefficient ranking, P[Formula: see text] and P[Formula: see text] were both higher than 0.99 for the 500 and 100 SNP genotypes, and higher than 0.98 for the 50 SNP genotypes. When parentage was assigned by a coefficient threshold, P[Formula: see text] was higher than 0.99 regardless of the number of SNPs, but P[Formula: see text] decreased from 0.99 (500 SNPs) to 0.97 (100 SNPs) and 0.92 (50 SNPs). If the sample of putative parents did not contain the true sire and parentage was rejected using a coefficient threshold, the algorithm achieved a P[Formula: see text] of 1 (500 SNPs), 0.99 (100 SNPs) and 0.97 (50 SNPs). The algorithm described here is easy to implement, fast and accurate, and is able to assign parentage using genomic marker data with a size as low as 50 SNPs.

  14. Improving removal-based estimates of abundance by sampling a population of spatially distinct subpopulations

    USGS Publications Warehouse

    Dorazio, R.M.; Jelks, H.L.; Jordan, F.

    2005-01-01

     A statistical modeling framework is described for estimating the abundances of spatially distinct subpopulations of animals surveyed using removal sampling. To illustrate this framework, hierarchical models are developed using the Poisson and negative-binomial distributions to model variation in abundance among subpopulations and using the beta distribution to model variation in capture probabilities. These models are fitted to the removal counts observed in a survey of a federally endangered fish species. The resulting estimates of abundance have similar or better precision than those computed using the conventional approach of analyzing the removal counts of each subpopulation separately. Extension of the hierarchical models to include spatial covariates of abundance is straightforward and may be used to identify important features of an animal's habitat or to predict the abundance of animals at unsampled locations.

  15. Sequential sampling: a novel method in farm animal welfare assessment.

    PubMed

    Heath, C A E; Main, D C J; Mullan, S; Haskell, M J; Browne, W J

    2016-02-01

    Lameness in dairy cows is an important welfare issue. As part of a welfare assessment, herd level lameness prevalence can be estimated from scoring a sample of animals, where higher levels of accuracy are associated with larger sample sizes. As the financial cost is related to the number of cows sampled, smaller samples are preferred. Sequential sampling schemes have been used for informing decision making in clinical trials. Sequential sampling involves taking samples in stages, where sampling can stop early depending on the estimated lameness prevalence. When welfare assessment is used for a pass/fail decision, a similar approach could be applied to reduce the overall sample size. The sampling schemes proposed here apply the principles of sequential sampling within a diagnostic testing framework. This study develops three sequential sampling schemes of increasing complexity to classify 80 fully assessed UK dairy farms, each with known lameness prevalence. Using the Welfare Quality herd-size-based sampling scheme, the first 'basic' scheme involves two sampling events. At the first sampling event half the Welfare Quality sample size is drawn, and then depending on the outcome, sampling either stops or is continued and the same number of animals is sampled again. In the second 'cautious' scheme, an adaptation is made to ensure that correctly classifying a farm as 'bad' is done with greater certainty. The third scheme is the only scheme to go beyond lameness as a binary measure and investigates the potential for increasing accuracy by incorporating the number of severely lame cows into the decision. The three schemes are evaluated with respect to accuracy and average sample size by running 100 000 simulations for each scheme, and a comparison is made with the fixed size Welfare Quality herd-size-based sampling scheme. All three schemes performed almost as well as the fixed size scheme but with much smaller average sample sizes. For the third scheme, an overall association between lameness prevalence and the proportion of lame cows that were severely lame on a farm was found. However, as this association was found to not be consistent across all farms, the sampling scheme did not prove to be as useful as expected. The preferred scheme was therefore the 'cautious' scheme for which a sampling protocol has also been developed.

  16. Detection of dermatophytes in healthy companion dogs and cats in eastern India.

    PubMed

    Debnath, C; Mitra, T; Kumar, A; Samanta, I

    2016-01-01

    In recent times increasing occurrence of dermatophytosis, especially among the school children in eastern India was evidenced along with increased tendency of keeping companion animals such as dogs and cats. This study was undertaken to detect the occurrence of dermatophytes with antifungal susceptibility among the companion animals. A total of 1501 healthy companion animals comprising 1209 dogs and 292 cats belonged to individual owners in and around Kolkata (West Bengal, India) were examined for the evidence of dermatophytosis during 2011-2013. The collected samples were subjected to direct examination by standard KOH mount technique. The samples were inoculated into both Sabouraud dextrose agar (SDA) with 0.05% chloramphenicol and 0.5% cycloheximide and dermatophyte test medium (DTM). Each of the fungal isolate was identified based upon its colony characteristics and hyphal and conidial cells it produced. Antifungal susceptibility of the isolates was tested by broth micro dilution assay using fluconazole, ketoconazole, itraconazole, miconazole, griseofulvin and amphotericin-B antifungals. Among the 1209 samples from dogs and 292 samples from cats, 253 (20.93%) and 109 (37.33%) samples were positive for dermatophytes by direct examination. Three identified species of dermatophytes with predominant occurrence were Microsporum canis, Microsporum gypseum and Trichophyton mentagrophytes. Ketoconazole (0.06-0.5 µgm/ml), itraconazole (0.03-0.5 µgm/ml) and amphotericin-B (0.03-0.5 µgm/ml) showed lowest MIC values against M. canis, T. mentagrophytes and M. gypseum, respectively. This is the first systemic report of dermatophytes in healthy companion animals with large numbers of samples in India.

  17. Diagnostic performance characteristics of a rapid field test for anthrax in cattle.

    PubMed

    Muller, Janine; Gwozdz, Jacek; Hodgeman, Rachel; Ainsworth, Catherine; Kluver, Patrick; Czarnecki, Jill; Warner, Simone; Fegan, Mark

    2015-07-01

    Although diagnosis of anthrax can be made in the field with a peripheral blood smear, and in the laboratory with bacterial culture or molecular based tests, these tests require either considerable experience or specialised equipment. Here we report on the evaluation of the diagnostic sensitivity and specificity of a simple and rapid in-field diagnostic test for anthrax, the anthrax immunochromatographic test (AICT). The AICT detects the protective antigen (PA) component of the anthrax toxin present within the blood of an animal that has died from anthrax. The test provides a result in 15min and offers the advantage of avoiding the necessity for on-site necropsy and subsequent occupational risks and environmental contamination. The specificity of the test was determined by testing samples taken from 622 animals, not infected with Bacillus anthracis. Diagnostic sensitivity was estimated on samples taken from 58 animals, naturally infected with B. anthracis collected over a 10-year period. All samples used to estimate the diagnostic sensitivity and specificity of the AICT were also tested using the gold standard of bacterial culture. The diagnostic specificity of the test was estimated to be 100% (99.4-100%; 95% CI) and the diagnostic sensitivity was estimated to be 93.1% (83.3-98.1%; 95% CI) (Clopper-Pearson method). Four samples produced false negative AICT results. These were among 9 samples, all of which tested positive for B. anthracis by culture, where there was a time delay between collection and testing of >48h and/or the samples were collected from animals that were >48h post-mortem. A statistically significant difference (P<0.001; Fishers exact test) was found between the ability of the AICT to detect PA in samples from culture positive animals <48h post-mortem, 49 of 49, Se=100% (92.8-100%; 95% CI) compared with samples tested >48h post-mortem 5 of 9 Se=56% (21-86.3%; 95% CI) (Clopper-Pearson method). Based upon these results a post hoc cut-off for use of the AICT of 48h post-mortem was applied, Se=100% (92.8-100%; 95% CI) and Sp=100% (99.4-100%; 95% CI). The high diagnostic sensitivity and specificity and the simplicity of the AICT enables it to be used for active surveillance in areas with a history of anthrax, or used as a preliminary tool in investigating sudden, unexplained death in cattle. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. 9 CFR 592.450 - Procedures for selecting appeal samples.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Procedures for selecting appeal samples. 592.450 Section 592.450 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION VOLUNTARY INSPECTION OF EGG PRODUCTS Appeals § 592.450 Procedures for selecting appeal samples. (a)...

  19. 9 CFR 592.450 - Procedures for selecting appeal samples.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Procedures for selecting appeal samples. 592.450 Section 592.450 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION VOLUNTARY INSPECTION OF EGG PRODUCTS Appeals § 592.450 Procedures for selecting appeal samples. (a)...

  20. 9 CFR 592.450 - Procedures for selecting appeal samples.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Procedures for selecting appeal samples. 592.450 Section 592.450 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION VOLUNTARY INSPECTION OF EGG PRODUCTS Appeals § 592.450 Procedures for selecting appeal samples. (a)...

  1. 9 CFR 592.450 - Procedures for selecting appeal samples.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Procedures for selecting appeal samples. 592.450 Section 592.450 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION VOLUNTARY INSPECTION OF EGG PRODUCTS Appeals § 592.450 Procedures for selecting appeal samples. (a)...

  2. [Formal sample size calculation and its limited validity in animal studies of medical basic research].

    PubMed

    Mayer, B; Muche, R

    2013-01-01

    Animal studies are highly relevant for basic medical research, although their usage is discussed controversially in public. Thus, an optimal sample size for these projects should be aimed at from a biometrical point of view. Statistical sample size calculation is usually the appropriate methodology in planning medical research projects. However, required information is often not valid or only available during the course of an animal experiment. This article critically discusses the validity of formal sample size calculation for animal studies. Within the discussion, some requirements are formulated to fundamentally regulate the process of sample size determination for animal experiments.

  3. A bacterial engineered glycoprotein as a novel antigen for diagnosis of bovine brucellosis.

    PubMed

    Ciocchini, Andrés E; Serantes, Diego A Rey; Melli, Luciano J; Guidolin, Leticia S; Iwashkiw, Jeremy A; Elena, Sebastián; Franco, Cristina; Nicola, Ana M; Feldman, Mario F; Comerci, Diego J; Ugalde, Juan E

    2014-08-27

    Brucellosis is a highly contagious zoonosis that affects livestock and human beings. Laboratory diagnosis of bovine brucellosis mainly relies on serological diagnosis using serum and/or milk samples. Although there are several serological tests with different diagnostic performance and capacity to differentiate vaccinated from infected animals, there is still no standardized reference antigen for the disease. Here we validate the first recombinant glycoprotein antigen, an N-formylperosamine O-polysaccharide-protein conjugate (OAg-AcrA), for diagnosis of bovine brucellosis. This antigen can be produced in homogeneous batches without the need of culturing pathogenic brucellae; all characteristics that make it appropriate for standardization. An indirect immunoassay based on the detection of anti O-polysaccharide IgG antibodies in bovine samples was developed coupling OAg-AcrA to magnetic beads or ELISA plates. As a proof of concept and to validate the antigen, we analyzed serum, whole blood and milk samples obtained from non-infected, experimentally infected and vaccinated animals included in a vaccination/infection trial performed in our laboratory as well as more than 1000 serum and milk samples obtained from naturally infected and S19-vaccinated animals from Argentina. Our results demonstrate that OAg-AcrA-based assays are highly accurate for diagnosis of bovine brucellosis, even in vaccinated herds, using different types of samples and in different platforms. We propose this novel recombinant glycoprotein as an antigen suitable for the development of new standard immunological tests for screening and confirmatory diagnosis of bovine brucellosis in regions or countries with brucellosis-control programs. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Detection of undeclared animal by-products in commercial canine canned foods: Comparative analyses by ELISA and PCR-RFLP coupled with slab gel electrophoresis or capillary gel electrophoresis.

    PubMed

    Hsieh, Ming-Kun; Shih, Pei-Yin; Wei, Chia-Fong; Vickroy, Thomas W; Chou, Chi-Chung

    2016-03-30

    The potential presence of undeclared animal by-products in pet foods is not subject to routine examination. Previously published methods for species-based identification of animal by-products have not been used routinely owing to inconsistent results. The present study evaluated the utility of several approaches for accurate identification of animal by-products in 11 commercial brands of canine canned foods. Canine canned foods from several countries were analysed by ELISA, PCR-RFLP coupled with slab-gel electrophoresis (SGE) and capillary gel electrophoresis (CGE) to test for evidence of by-products derived from cattle, chicken, sheep or pig. While CGE-based analysis detected all (24) animal-derived by-products that were reported for the 11 test samples, SGE and ELISA detected only 22/24 (92%) and 14/24 (58%) of labelled by-products, respectively. In addition, undeclared animal by-products were found using all three analytical approaches with CGE detecting more positives (19) than SGE (17) or ELISA (5). Significant disparities were evident between the labelled contents and the detected content of animal by-products. CGE-based testing for PCR products appears to provide greater sensitivity and accuracy than either SGE or ELISA-based methods. As testing of commercial products becomes more reliable and mainstream, manufacturers will need to develop more thorough and accurate labelling protocols. © 2015 Society of Chemical Industry.

  5. Considering aspects of the 3Rs principles within experimental animal biology.

    PubMed

    Sneddon, Lynne U; Halsey, Lewis G; Bury, Nic R

    2017-09-01

    The 3Rs - Replacement, Reduction and Refinement - are embedded into the legislation and guidelines governing the ethics of animal use in experiments. Here, we consider the advantages of adopting key aspects of the 3Rs into experimental biology, represented mainly by the fields of animal behaviour, neurobiology, physiology, toxicology and biomechanics. Replacing protected animals with less sentient forms or species, cells, tissues or computer modelling approaches has been broadly successful. However, many studies investigate specific models that exhibit a particular adaptation, or a species that is a target for conservation, such that their replacement is inappropriate. Regardless of the species used, refining procedures to ensure the health and well-being of animals prior to and during experiments is crucial for the integrity of the results and legitimacy of the science. Although the concepts of health and welfare are developed for model organisms, relatively little is known regarding non-traditional species that may be more ecologically relevant. Studies should reduce the number of experimental animals by employing the minimum suitable sample size. This is often calculated using power analyses, which is associated with making statistical inferences based on the P -value, yet P -values often leave scientists on shaky ground. We endorse focusing on effect sizes accompanied by confidence intervals as a more appropriate means of interpreting data; in turn, sample size could be calculated based on effect size precision. Ultimately, the appropriate employment of the 3Rs principles in experimental biology empowers scientists in justifying their research, and results in higher-quality science. © 2017. Published by The Company of Biologists Ltd.

  6. Rope-based oral fluid sampling for early detection of classical swine fever in domestic pigs at group level.

    PubMed

    Dietze, Klaas; Tucakov, Anna; Engel, Tatjana; Wirtz, Sabine; Depner, Klaus; Globig, Anja; Kammerer, Robert; Mouchantat, Susan

    2017-01-05

    Non-invasive sampling techniques based on the analysis of oral fluid specimen have gained substantial importance in the field of swine herd management. Methodological advances have a focus on endemic viral diseases in commercial pig production. More recently, these approaches have been adapted to non-invasive sampling of wild boar for transboundary animal disease detection for which these effective population level sampling methods have not been available. In this study, a rope-in-a-bait based oral fluid sampling technique was tested to detect classical swine fever virus nucleic acid shedding from experimentally infected domestic pigs. Separated in two groups treated identically, the course of the infection was slightly differing in terms of onset of the clinical signs and levels of viral ribonucleic acid detection in the blood and oral fluid. The technique was capable of detecting classical swine fever virus nucleic acid as of day 7 post infection coinciding with the first detection in conventional oropharyngeal swab samples from some individual animals. Except for day 7 post infection in the "slower onset group", the chances of classical swine fever virus nucleic acid detection in ropes were identical or higher as compared to the individual sampling. With the provided evidence, non-invasive oral fluid sampling at group level can be considered as additional cost-effective detection tool in classical swine fever prevention and control strategies. The proposed methodology is of particular use in production systems with reduced access to veterinary services such as backyard or scavenging pig production where it can be integrated in feeding or baiting practices.

  7. Serum concentration comparisons of amino acids, fatty acids, lipoproteins, vitamins A and E, and minerals between zoo and free-ranging giraffes (Giraffa camelopardalis).

    PubMed

    Schmidt, Debra A; Koutsos, Elizabeth A; Ellersieck, Mark R; Griffin, Mark E

    2009-03-01

    Serum concentrations of amino acids, fatty acids, lipoproteins, vitamins A and E, and minerals in zoo giraffes (Giraffa camelopardalis) were compared to values obtained from free-ranging giraffes in an effort to identify potential nutritional differences in the zoo population. Zoo giraffes have a specific set of maladies that may be nutritionally related, including peracute mortality, energy malnutrition, pancreatic disease, urolithiasis, hoof disease, and severe intestinal parasitism. Dietary requirements for giraffes are not known; invasive studies used with domestic animals cannot be performed on zoo animals. Though domestic animal standards are often used to evaluate nutritional health of exotic animals, they may not be the most appropriate standards to use. Serum samples from 20 zoo giraffes at 10 zoological institutions in the United States were compared to previously collected samples from 24 free-ranging giraffes in South Africa. Thirteen of the zoo animal samples were collected from animals trained for blood collection, and seven were banked samples obtained from a previous serum collection. Dietary information was also collected on each zoo giraffe; most zoo giraffe diets consisted of alfalfa-based pellets (acid detergent fiber-16), alfalfa hay, and browse in varying quantities. Differences between zoo and free-ranging giraffes, males and females, and adults and subadults were analyzed with the use of a 2 x 2 x 2 factorial and Fisher's Least Significant Difference (LSD) for mean separation. Of the 84 parameters measured, 54 (60%) were significantly different (P < or = 0.05) between zoo and free-ranging giraffes. Nine (11%) items were significantly different (P < or = 0.05) between adult and subadult animals. Only one parameter, sodium concentration, was found to be significantly different (P < or = 0.05) between genders. Further investigation in zoo giraffe diets is needed to address the differences seen in this study and the potentially related health problems.

  8. Molecular and serological study of rickettsial infection in humans, and in wild and farm animals, in the province of Burgos, Spain.

    PubMed

    Lledó, Lourdes; Domínguez-Peñafiel, Gerardo; Giménez-Pardo, Consuelo; Gegúndez, Isabel; González, Rosario; Saz, José Vicente

    2014-06-01

    Limited information is available on the presence of rickettsial infection in humans and animal reservoirs in Spain. Exposure to spotted fever group rickettsia in healthy humans and in farm and wild animals in the Province of Burgos, Spain, was examined by serological methods. Rickettsial DNA was also sought by PCR in animal samples. Of 102 human serum samples examined by indirect immunofluorescence assays (IFA), 5.88% were positive for antibodies against Rickettsia conorii (titers 1/128-1/512). Significant differences were detected in human seroprevalence with respect to age. In further IFAs, 102 out of 375 (27.2%) serum samples from the wild animals reacted with R. conorii antigens (titers 1/64-1/1024); 32 out of 281 (11.38%) samples from farm animals were also positive for R. conorii (titers 1/64-1/2048). The prevalence detected among total wild animals was significantly higher than among total farm animals. No rickettsial DNA was found by PCR in any farm or wild animal sample.

  9. 9 CFR 312.10 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Official mark for maintaining the identity and integrity of samples. 312.10 Section 312.10 Animals and Animal Products FOOD SAFETY AND... of the Federal Meat Inspection Act shall bear the designation “Sample Seal” accompanied by the...

  10. 9 CFR 381.112 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Official mark for maintaining the identity and integrity of samples. 381.112 Section 381.112 Animals and Animal Products FOOD SAFETY AND... Inspection Act shall bear the designation “Sample Seal” accompanied by the official USDA logo as shown below...

  11. 9 CFR 318.9 - Samples of products, water, dyes, chemicals, etc., to be taken for examination.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Samples of products, water, dyes, chemicals, etc., to be taken for examination. 318.9 Section 318.9 Animals and Animal Products FOOD SAFETY... ESTABLISHMENTS; REINSPECTION AND PREPARATION OF PRODUCTS General § 318.9 Samples of products, water, dyes...

  12. 9 CFR 318.9 - Samples of products, water, dyes, chemicals, etc., to be taken for examination.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Samples of products, water, dyes, chemicals, etc., to be taken for examination. 318.9 Section 318.9 Animals and Animal Products FOOD SAFETY... ESTABLISHMENTS; REINSPECTION AND PREPARATION OF PRODUCTS General § 318.9 Samples of products, water, dyes...

  13. 9 CFR 318.9 - Samples of products, water, dyes, chemicals, etc., to be taken for examination.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Samples of products, water, dyes, chemicals, etc., to be taken for examination. 318.9 Section 318.9 Animals and Animal Products FOOD SAFETY... ESTABLISHMENTS; REINSPECTION AND PREPARATION OF PRODUCTS General § 318.9 Samples of products, water, dyes...

  14. 9 CFR 318.9 - Samples of products, water, dyes, chemicals, etc., to be taken for examination.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Samples of products, water, dyes, chemicals, etc., to be taken for examination. 318.9 Section 318.9 Animals and Animal Products FOOD SAFETY... ESTABLISHMENTS; REINSPECTION AND PREPARATION OF PRODUCTS General § 318.9 Samples of products, water, dyes...

  15. 9 CFR 312.10 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Official mark for maintaining the identity and integrity of samples. 312.10 Section 312.10 Animals and Animal Products FOOD SAFETY AND... § 312.10 Official mark for maintaining the identity and integrity of samples. The official mark for use...

  16. 9 CFR 381.112 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Official mark for maintaining the identity and integrity of samples. 381.112 Section 381.112 Animals and Animal Products FOOD SAFETY AND... mark for maintaining the identity and integrity of samples. The official mark for use in sealing...

  17. 9 CFR 312.10 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Official mark for maintaining the identity and integrity of samples. 312.10 Section 312.10 Animals and Animal Products FOOD SAFETY AND... § 312.10 Official mark for maintaining the identity and integrity of samples. The official mark for use...

  18. 9 CFR 381.112 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Official mark for maintaining the identity and integrity of samples. 381.112 Section 381.112 Animals and Animal Products FOOD SAFETY AND... mark for maintaining the identity and integrity of samples. The official mark for use in sealing...

  19. 9 CFR 381.112 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Official mark for maintaining the identity and integrity of samples. 381.112 Section 381.112 Animals and Animal Products FOOD SAFETY AND... mark for maintaining the identity and integrity of samples. The official mark for use in sealing...

  20. 9 CFR 312.10 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Official mark for maintaining the identity and integrity of samples. 312.10 Section 312.10 Animals and Animal Products FOOD SAFETY AND... § 312.10 Official mark for maintaining the identity and integrity of samples. The official mark for use...

  1. 9 CFR 312.10 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Official mark for maintaining the identity and integrity of samples. 312.10 Section 312.10 Animals and Animal Products FOOD SAFETY AND... § 312.10 Official mark for maintaining the identity and integrity of samples. The official mark for use...

  2. 9 CFR 381.112 - Official mark for maintaining the identity and integrity of samples.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Official mark for maintaining the identity and integrity of samples. 381.112 Section 381.112 Animals and Animal Products FOOD SAFETY AND... mark for maintaining the identity and integrity of samples. The official mark for use in sealing...

  3. 9 CFR 318.9 - Samples of products, water, dyes, chemicals, etc., to be taken for examination.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Samples of products, water, dyes, chemicals, etc., to be taken for examination. 318.9 Section 318.9 Animals and Animal Products FOOD SAFETY... ESTABLISHMENTS; REINSPECTION AND PREPARATION OF PRODUCTS General § 318.9 Samples of products, water, dyes...

  4. Prevalence and distribution of paratuberculosis (Johne's disease) in cattle herds in Ireland

    PubMed Central

    2009-01-01

    A simple random survey was conducted in Ireland during 2005 to estimate the ELISA-prevalence of paratuberculosis, commonly called Johne's disease (JD), in the cattle population. Serum samples were collected from all 20,322 females/breeding bulls over 12 months-of-age in 639 herds. All samples were tested using a commercially available absorbed ELISA. The overall prevalence of infected herds, based on the presence of at least one ELISA-positive animal, was 21.4% (95% CI 18.4%-24.9%). Herd prevalence levels amongst dairy herds (mean 31.5%; 95% CI: 24.6%, 39.3%) was higher than among beef herds (mean 17.9%; 95% CI: 14.6%-21.8%). However, the animal level prevalence was similar. The true prevalence among all animals tested, was calculated to be 2.86% (95%CI: 2.76, 2.97) and for animals >= 2 yrs, it was 3.30% (95%CI: 3.17, 3.43). For animals in beef herds, true prevalence was 3.09% (95%CI: 2.93, 3.24), and for those in dairy herds, 2.74% (95%CI: 2.59, 2.90). The majority of herds had only one ELISA-positive infected animal. Only 6.4% (95% CI 4.7%-8.7%) of all herds had more than one ELISA-positive infected animal; 13.3% (CI 8.7%-19.7%) of dairy herds ranging from two to eight ELISA-positive infected animals; and, 3.9% beef herds (CI 2.4%-6.2%) ranging from two to five ELISA-positive infected animals. The true prevalence of herds infected and shedding Mycobacterium avium subspecies paratuberculosis is estimated to be 9.5% for all herd types; 20.6% for dairy herds; and 7.6% for beef herds. If ELISA positive animals <2-years-of-age are excluded, the true herd prevalene reduces to: 9.3% for all herd types; 19.6% for dairy herds; and 6.3% for beef herds based on a test specificity (Sp) of 99.8% and test sensitivity (Se) (i.e., ability to detect culture-positive, infected animals shedding at any level) of 27.8-28.9%. PMID:21851740

  5. Real-time quantitative fluorescence measurement of microscale cell culture analog systems

    NASA Astrophysics Data System (ADS)

    Oh, Taek-il; Kim, Donghyun; Tatosian, Daniel; Sung, Jong Hwan; Shuler, Michael

    2007-02-01

    A microscale cell culture analog (μCCA) is a cell-based lab-on-a-chip assay that, as an animal surrogate, is applied to pharmacological studies for toxicology tests. A μCCA typically comprises multiple chambers and microfluidics that connect the chambers, which represent animal organs and blood flow to mimic animal metabolism more realistically. A μCCA is expected to provide a tool for high-throughput drug discovery. Previously, a portable fluorescence detection system was investigated for a single μCCA device in real-time. In this study, we present a fluorescence-based imaging system that provides quantitative real-time data of the metabolic interactions in μCCAs with an emphasis on measuring multiple μCCA samples simultaneously for high-throughput screening. The detection system is based on discrete optics components, with a high-power LED and a charge-coupled device (CCD) camera as a light source and a detector, for monitoring cellular status on the chambers of each μCCA sample. Multiple samples are characterized mechanically on a motorized linear stage, which is fully-automated. Each μCCA sample has four chambers, where cell lines MES-SA/DX- 5, and MES-SA (tumor cells of human uterus) have been cultured. All cell-lines have been transfected to express the fusion protein H2B-GFP, which is a human histone protein fused at the amino terminus to EGFP. As a model cytotoxic drug, 10 μM doxorubicin (DOX) was used. Real-time quantitative data of the intensity loss of enhanced green fluorescent protein (EGFP) during cell death of target cells have been collected over several minutes to 40 hours. Design issues and improvements are also discussed.

  6. Using dried blood spot sampling to improve data quality and reduce animal use in mouse pharmacokinetic studies.

    PubMed

    Wickremsinhe, Enaksha R; Perkins, Everett J

    2015-03-01

    Traditional pharmacokinetic analysis in nonclinical studies is based on the concentration of a test compound in plasma and requires approximately 100 to 200 μL blood collected per time point. However, the total blood volume of mice limits the number of samples that can be collected from an individual animal-often to a single collection per mouse-thus necessitating dosing multiple mice to generate a pharmacokinetic profile in a sparse-sampling design. Compared with traditional methods, dried blood spot (DBS) analysis requires smaller volumes of blood (15 to 20 μL), thus supporting serial blood sampling and the generation of a complete pharmacokinetic profile from a single mouse. Here we compare plasma-derived data with DBS-derived data, explain how to adopt DBS sampling to support discovery mouse studies, and describe how to generate pharmacokinetic and pharmacodynamic data from a single mouse. Executing novel study designs that use DBS enhances the ability to identify and streamline better drug candidates during drug discovery. Implementing DBS sampling can reduce the number of mice needed in a drug discovery program. In addition, the simplicity of DBS sampling and the smaller numbers of mice needed translate to decreased study costs. Overall, DBS sampling is consistent with 3Rs principles by achieving reductions in the number of animals used, decreased restraint-associated stress, improved data quality, direct comparison of interanimal variability, and the generation of multiple endpoints from a single study.

  7. Prevalence and molecular identification of Chlamydia abortus in commercial dairy goat farms in a hot region in Mexico.

    PubMed

    Campos-Hernández, Eleuterio; Vázquez-Chagoyán, Juan Carlos; Salem, Abdelfattah Z M; Saltijeral-Oaxaca, Jorge Antonio; Escalante-Ochoa, Cristina; López-Heydeck, Sandra M; de Oca-Jiménez, Roberto Montes

    2014-08-01

    The aim of this study was to determine the seroprevalence and presence of Chlamydia abortus in Saanen breed female goats from commercial dairy goat farms under intensive production in the municipality of Guanajuato, Mexico. Sera were collected to determine the prevalence of anti-C. abortus IgG antibodies using recombinant enzyme-linked immunosorbent assay (rELISA) and cell culture. Polymerase chain reaction (PCR) was used to prove the presence of the pathogen in swab samples collected from the vagina and rectum of selected animals. Additionally, foetal tissue samples from a sudden abortion were collected. C. abortus prevalence in female goats of commercial milking farms sampled in Guanajuato, Mexico, was 4.87% (n = 246). Seropositive animals were found in six out of nine (66.6%) dairy goat farms sampled, and prevalence among animals in individual farms ranged between 3.44 and 13.51%. C. abortus was detected using PCR in spleen tissue from the aborted foetus. PCR-based detection, as well as isolation from vaginal and rectal swabs, was not possible in the present study. Isolation through cell culture was also unsuccessful from aborted foetal tissue samples. In conclusion, the results from rELISA and PCR show that C. abortus is present in dairy goat farms in the state of Guanajuato, Mexico.

  8. Animal ecology meets GPS-based radiotelemetry: a perfect storm of opportunities and challenges

    PubMed Central

    Cagnacci, Francesca; Boitani, Luigi; Powell, Roger A.; Boyce, Mark S.

    2010-01-01

    Global positioning system (GPS) telemetry technology allows us to monitor and to map the details of animal movement, securing vast quantities of such data even for highly cryptic organisms. We envision an exciting synergy between animal ecology and GPS-based radiotelemetry, as for other examples of new technologies stimulating rapid conceptual advances, where research opportunities have been paralleled by technical and analytical challenges. Animal positions provide the elemental unit of movement paths and show where individuals interact with the ecosystems around them. We discuss how knowing where animals go can help scientists in their search for a mechanistic understanding of key concepts of animal ecology, including resource use, home range and dispersal, and population dynamics. It is probable that in the not-so-distant future, intense sampling of movements coupled with detailed information on habitat features at a variety of scales will allow us to represent an animal's cognitive map of its environment, and the intimate relationship between behaviour and fitness. An extended use of these data over long periods of time and over large spatial scales can provide robust inferences for complex, multi-factorial phenomena, such as meta-analyses of the effects of climate change on animal behaviour and distribution. PMID:20566493

  9. Comparison of PCR-Based Diagnosis with Centrifuged-Based Enrichment Method for Detection of Borrelia persica in Animal Blood Samples.

    PubMed

    Naddaf, S R; Kishdehi, M; Siavashi, Mr

    2011-01-01

    The mainstay of diagnosis of relapsing fever (RF) is demonstration of the spirochetes in Giemsa-stained thick blood smears, but during non fever periods the bacteria are very scanty and rarely detected in blood smears by microscopy. This study is aimed to evaluate the sensitivity of different methods developed for detection of low-grade spirochetemia. Animal blood samples with low degrees of spirochetemia were tested with two PCRs and a nested PCR targeting flaB, GlpQ, and rrs genes. Also, a centrifuged-based enrichment method and Giemsa staining were performed on blood samples with various degrees of spirochetemia. The flaB-PCR and nested rrs-PCR turned positive with various degrees of spirochetemia including the blood samples that turned negative with dark-field microscopy. The GlpQ-PCR was positive as far as at least one spirochete was seen in 5-10 microscopic fields. The sensitivity of GlpQ-PCR increased when DNA from Buffy Coat Layer (BCL) was used as template. The centrifuged-based enrichment method turned positive with as low concentration as 50 bacteria/ml blood, while Giemsa thick staining detected bacteria with concentrations ≥ 25000 bacteria/ml. Centrifuged-based enrichment method appeared as much as 500-fold more sensitive than thick smears, which makes it even superior to some PCR assays. Due to simplicity and minimal laboratory requirements, this method can be considered a valuable tool for diagnosis of RF in rural health centers.

  10. Reduction of Sample Size Requirements by Bilateral Versus Unilateral Research Designs in Animal Models for Cartilage Tissue Engineering

    PubMed Central

    Orth, Patrick; Zurakowski, David; Alini, Mauro; Cucchiarini, Magali

    2013-01-01

    Advanced tissue engineering approaches for articular cartilage repair in the knee joint rely on translational animal models. In these investigations, cartilage defects may be established either in one joint (unilateral design) or in both joints of the same animal (bilateral design). We hypothesized that a lower intraindividual variability following the bilateral strategy would reduce the number of required joints. Standardized osteochondral defects were created in the trochlear groove of 18 rabbits. In 12 animals, defects were produced unilaterally (unilateral design; n=12 defects), while defects were created bilaterally in 6 animals (bilateral design; n=12 defects). After 3 weeks, osteochondral repair was evaluated histologically applying an established grading system. Based on intra- and interindividual variabilities, required sample sizes for the detection of discrete differences in the histological score were determined for both study designs (α=0.05, β=0.20). Coefficients of variation (%CV) of the total histological score values were 1.9-fold increased following the unilateral design when compared with the bilateral approach (26 versus 14%CV). The resulting numbers of joints needed to treat were always higher for the unilateral design, resulting in an up to 3.9-fold increase in the required number of experimental animals. This effect was most pronounced for the detection of small-effect sizes and estimating large standard deviations. The data underline the possible benefit of bilateral study designs for the decrease of sample size requirements for certain investigations in articular cartilage research. These findings might also be transferred to other scoring systems, defect types, or translational animal models in the field of cartilage tissue engineering. PMID:23510128

  11. Monitoring Method of Cow Anthrax Based on Gis and Spatial Statistical Analysis

    NASA Astrophysics Data System (ADS)

    Li, Lin; Yang, Yong; Wang, Hongbin; Dong, Jing; Zhao, Yujun; He, Jianbin; Fan, Honggang

    Geographic information system (GIS) is a computer application system, which possesses the ability of manipulating spatial information and has been used in many fields related with the spatial information management. Many methods and models have been established for analyzing animal diseases distribution models and temporal-spatial transmission models. Great benefits have been gained from the application of GIS in animal disease epidemiology. GIS is now a very important tool in animal disease epidemiological research. Spatial analysis function of GIS can be widened and strengthened by using spatial statistical analysis, allowing for the deeper exploration, analysis, manipulation and interpretation of spatial pattern and spatial correlation of the animal disease. In this paper, we analyzed the cow anthrax spatial distribution characteristics in the target district A (due to the secret of epidemic data we call it district A) based on the established GIS of the cow anthrax in this district in combination of spatial statistical analysis and GIS. The Cow anthrax is biogeochemical disease, and its geographical distribution is related closely to the environmental factors of habitats and has some spatial characteristics, and therefore the correct analysis of the spatial distribution of anthrax cow for monitoring and the prevention and control of anthrax has a very important role. However, the application of classic statistical methods in some areas is very difficult because of the pastoral nomadic context. The high mobility of livestock and the lack of enough suitable sampling for the some of the difficulties in monitoring currently make it nearly impossible to apply rigorous random sampling methods. It is thus necessary to develop an alternative sampling method, which could overcome the lack of sampling and meet the requirements for randomness. The GIS computer application software ArcGIS9.1 was used to overcome the lack of data of sampling sites.Using ArcGIS 9.1 and GEODA to analyze the cow anthrax spatial distribution of district A. we gained some conclusions about cow anthrax' density: (1) there is a spatial clustering model. (2) there is an intensely spatial autocorrelation. We established a prediction model to estimate the anthrax distribution based on the spatial characteristic of the density of cow anthrax. Comparing with the true distribution, the prediction model has a well coincidence and is feasible to the application. The method using a GIS tool facilitates can be implemented significantly in the cow anthrax monitoring and investigation, and the space statistics - related prediction model provides a fundamental use for other study on space-related animal diseases.

  12. Cattle mortality in the Thane district, India: a study of cause/effect relationships.

    PubMed

    Dogra, R K; Murthy, R C; Srivastava, A K; Gaur, J S; Shukla, L J; Varmani, B M

    1996-02-01

    An unexpected mortality of more than 300 cattle was investigated near a metal recovery factory located in a rural area of the Thane district of India. The factory was engaged in reclaiming lead, aluminum, tin, and zinc from discarded lead storage batteries and soft drink cans. The environmental samples (soil, leaves, grass, slag, water, and sediment), human blood and hair and animal samples (blood, urine, peritoneal fluid, liver, kidney, cow dung, ribs, and femur), collected for analysis revealed toxic levels of lead, cadmium, and chromium. Clinical examination of factory workers and school children revealed cough, fever, gastric problems, abdominal pain, skin lesions (scabies), and blue line on gums. Histopathological examination of animal tissues revealed chronic pathology with lead inclusion bodies in hepatocytes and renal tubules. Based on environmental, clinical, analytical, and histopathological observations, the mortality has been attributed to toxic levels of metals in the body and the malnourished status of the animals.

  13. Detection and phylogenetic analysis of a new adenoviral polymerase gene in reptiles in Korea.

    PubMed

    Bak, Eun-Jung; Jho, Yeonsook; Woo, Gye-Hyeong

    2018-06-01

    Over a period of 7 years (2004-2011), samples from 34 diseased reptiles provided by local governments, zoos, and pet shops were tested for viral infection. Animals were diagnosed based on clinical signs, including loss of appetite, diarrhea, rhinorrhea, and unexpected sudden death. Most of the exotic animals had gastrointestinal problems, such as mucosal redness and ulcers, while the native animals had no clinical symptoms. Viral sequences were found in seven animals. Retroviral genes were amplified from samples from five Burmese pythons (Python molurus bivittatus), an adenovirus was detected in a panther chameleon (Furcifer pardalis), and an adenovirus and a paramyxovirus were detected in a tropical girdled lizard (Cordylus tropidosternum). Phylogenetic analysis of retroviruses and paramyxoviruses showed the highest sequence identity to both a Python molurus endogenous retrovirus and a Python curtus endogenous retrovirus and to a lizard isolate, respectively. Partial sequencing of an adenoviral DNA polymerase gene from the lizard isolate suggested that the corresponding virus was a novel isolate different from the reference strain (accession no. AY576677.1). The virus was not isolated but was detected, using molecular genetic techniques, in a lizard raised in a pet shop. This animal was also coinfected with a paramyxovirus.

  14. Evaluation of PCDD/Fs characterization in animal feed and feed additives.

    PubMed

    Kim, MeeKyung; Kim, Sooyeon; Yun, Seon Jong; Kwon, Jin-Wook; Son, Seong-Wan

    2007-09-01

    Safety control of feed and feed additives is necessary to have safe food of animal origin. Based on media reports, nine incidents regarding dioxins and/or PCBs contaminations occurred worldwide during the last decade. Korea is a country which imports feed and feed additives. In this study, various kinds of feed and feed additives were analyzed to monitor the contamination level of dioxins. The level of PCDD/Fs in fish oil was the highest with a concentration of 23.33ngkg(-1), which is equivalent to a toxicological concentration of 4.68ngWHO-TEQ/kg. Feed from animals origin such as chicken meal, animal fat, fish meal, fish oil, and shell powder showed relatively higher concentrations of PCDD/Fs. Feed from plants origin, minerals, and additives ranged from non-detects for bit pulp and ethoxyquin to 8.28ngkg(-1) for dl-methionine. From a toxicological point of view, the highest concentration in vitamins was 0.08ngWHO-TEQ/kg among the feed additives. 2,3,4,7,8-PeCDF was the dominant congener in samples of fish oil, fish meal, and shell powder. Animal fat showed that the pattern of PCDD/Fs depends on the sources of contamination. A sample of animal fat showed 1,2,3,4,7,8-HxCDF and the other sample showed 1,2,3,4,7,8-HxCDD as a primary congener. Generally, low levels of PCDDs were detected in feed additives. Patterns of PCDD/Fs in choline chloride were different with that in choline chloride from an incident in Europe in 2000.

  15. Risk-Based Sampling: I Don't Want to Weight in Vain.

    PubMed

    Powell, Mark R

    2015-12-01

    Recently, there has been considerable interest in developing risk-based sampling for food safety and animal and plant health for efficient allocation of inspection and surveillance resources. The problem of risk-based sampling allocation presents a challenge similar to financial portfolio analysis. Markowitz (1952) laid the foundation for modern portfolio theory based on mean-variance optimization. However, a persistent challenge in implementing portfolio optimization is the problem of estimation error, leading to false "optimal" portfolios and unstable asset weights. In some cases, portfolio diversification based on simple heuristics (e.g., equal allocation) has better out-of-sample performance than complex portfolio optimization methods due to estimation uncertainty. Even for portfolios with a modest number of assets, the estimation window required for true optimization may imply an implausibly long stationary period. The implications for risk-based sampling are illustrated by a simple simulation model of lot inspection for a small, heterogeneous group of producers. © 2015 Society for Risk Analysis.

  16. Bluetongue sentinel surveillance program and cross-sectional serological survey in cattle in Belgium in 2010-2011.

    PubMed

    Vangeel, I; De Leeuw, I; Méroc, E; Vandenbussche, F; Riocreux, F; Hooyberghs, J; Raemaekers, M; Houdart, P; Van der Stede, Y; De Clercq, K

    2012-10-01

    Bluetongue virus serotype 8 (BTV-8) emerged in Central Western Europe in 2006 causing a large scale epidemic in 2007 that involved several European Union (EU) countries including Belgium. As in several other EU member states, vaccination against BTV-8 with inactivated vaccines was initiated in Belgium in spring 2008 and appeared to be successful. Since 2009, no clinical cases of Bluetongue (BT) have been reported in Belgium and BTV-8 circulation seemed to have completely disappeared by spring 2010. Therefore, a series of repeated cross-sectional surveys, the BT sentinel surveillance program, based on virus detection in blood samples by means of real-time RT-PCR (RT-qPCR) were carried out in dairy cattle from the end of 2010 onwards with the aim to demonstrate the absence of BTV circulation in Belgium. This paper describes the results of the first two sampling rounds of this BT sentinel surveillance program carried out in October-November 2010 and January-February 2011. In addition, the level of BTV-specific maternal antibodies in young non-vaccinated animals was monitored and the level of herd immunity against BTV-8 after 3 consecutive years of compulsory BTV-8 vaccination was measured by ELISA. During the 1st sampling round of the BT sentinel surveillance program, 15 animals tested positive and 2 animals tested doubtful for BTV RNA by RT-qPCR. During the 2nd round, 17 animals tested positive and 5 animals tested doubtful. The positive/doubtful animals in both rounds were re-sampled 2-4 weeks after the original sampling and then all tested negative by RT-qPCR. These results demonstrate the absence of BTV circulation in Belgium in 2010 at a minimum expected prevalence of 2% and 95% confidence level. The study of the maternal antibodies in non-vaccinated animals showed that by the age of 7 months maternal antibodies against BTV had disappeared in most animals. The BTV seroprevalence at herd level after 3 years of compulsory BTV-8 vaccination was very high (97.4% [95% CI: 96.2-98.2]). The overall true within-herd BTV seroprevalence in 6-24 month old Belgian cattle in early 2011 was estimated at 73.4% (95% CI: 71.3-75.4). Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Development, validation and application of an ICP-MS/MS method to quantify minerals and (ultra-)trace elements in human serum.

    PubMed

    Meyer, Sören; Markova, Mariya; Pohl, Gabriele; Marschall, Talke A; Pivovarova, Olga; Pfeiffer, Andreas F H; Schwerdtle, Tanja

    2018-09-01

    Multi-element determination in human samples is very challenging. Especially in human intervention studies sample volumes are often limited to a few microliters and due to the high number of samples a high-throughput is indispensable. Here, we present a state-of-the-art ICP-MS/MS-based method for the analysis of essential (trace) elements, namely Mg, Ca, Fe, Cu, Zn, Mo, Se and I, as well as food-relevant toxic elements such as As and Cd. The developed method was validated regarding linearity of the calibration curves, method LODs and LOQs, selectivity and trueness as well as precision. The established reliable method was applied to quantify the element serum concentrations of participants of a human intervention study (LeguAN). The participants received isocaloric diets, either rich in plant protein or in animal protein. While the serum concentrations of Mg and Mo increased in participants receiving the plant protein-based diet (above all legumes), the Se concentration in serum decreased. In contrast, the animal protein-based diet, rich in meat and dairy products, resulted in an increased Se concentration in serum. Copyright © 2018 Elsevier GmbH. All rights reserved.

  18. Survey and Experimental Infection of Enteropathogenic Escherichia coli in Common Marmosets (Callithrix jacchus).

    PubMed

    Hayashimoto, Nobuhito; Inoue, Takashi; Morita, Hanako; Yasuda, Masahiko; Ueno, Masami; Kawai, Kenji; Itoh, Toshio

    2016-01-01

    Common marmosets (Callithrix jacchus) are frequently used for biomedical research but can be afflicted with diarrhea-a serious and potentially lethal health problem. Enteropathogenic Escherichia coli (EPEC) is thought to be the causative pathogen of hemorrhagic typhlocolitis in common marmosets, but the actual incidence of the disease and the relationship between EPEC and hematochezia are unknown. This study investigated the prevalence of EPEC infection in common marmosets and the association between EPEC and hematochezia. A total of 230 stool or rectal swab samples were collected from 230 common marmosets (98 clinically healthy, 85 diarrhea, and 47 bloody stool samples) and tested by culture-based detection and PCR amplification of VT1, VT2, LT, ST, eae, and bfp genes. Healthy animals were divided into three groups (n = 4 each for high and low concentration groups and n = 2 as negative control), and those in the experimental groups were perorally inoculated with a 2-ml of suspension of EPEC R811 strain adjusted to 5 × 108 (high concentration) and 5 × 104 (low concentration) CFU/ ml. Two animals in each group were examined 3 and 14 days post-inoculation (DPI). EPEC was detected in 10 of 98 clinically healthy samples (10.2%), 17 of 85 diarrhea samples (20%), and all 47 bloody stool samples (100%), with a significant difference detected between presence of EPEC and sample status (P < 0.01). Acute hematochezia was observed in all animals of the high-concentration group but not in other groups at 1 or 2 DPI. A histopathological examination revealed the attachment of gram-negative bacilli to epithelial apical membranes and desquamated epithelial cells in the cecum of animals in the high-concentration group at 3 DPI. These findings suggest that EPEC is a causative agent of hemorrhagic typhlocolitis in common marmosets.

  19. Examining the relationship between childhood animal cruelty motives and recurrent adult violent crimes toward humans.

    PubMed

    Overton, Joshua C; Hensley, Christopher; Tallichet, Suzanne E

    2012-03-01

    Few researchers have studied the predictive ability of childhood animal cruelty motives as they are associated with later recurrent violence toward humans. Based on a sample of 180 inmates at one medium- and one maximum-security prison in a Southern state, the present study examines the relationship among several retrospectively identified motives (fun, out of anger, hate for the animal, and imitation) for childhood animal cruelty and the later commission of violent crimes (murder, rape, assault, and robbery) against humans. Almost two thirds of the inmates reported engaging in childhood animal cruelty for fun, whereas almost one fourth reported being motivated either out of anger or imitation. Only one fifth of the respondents reported they had committed acts of animal cruelty because they hated the animal. Regression analyses revealed that recurrent animal cruelty was the only statistically significant variable in the model. Respondents who had committed recurrent childhood animal cruelty were more likely to have had committed recurrent adult violence toward humans. None of the motives for committing childhood animal cruelty had any effect on later violence against humans.

  20. Detecting quinoxaline-2-carboxylic acid in animal tissues by using sensitive rapid enzyme-linked immunosorbent assay and time-resolved fluoroimmunoassay.

    PubMed

    Le, Tao; Yu, Huan; Niu, Xiaodong

    2015-05-15

    An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and time-resolved fluoroimmunoassay (TR-FIA) based on an anti-N-butylquinoxaline-2-carboxamide (BQCA) monoclonal antibody were standardized and validated for quinoxaline-2-carboxylic acid (QCA) screening in animal tissues and its performance were compared to HPLC. The sensitivities obtained for edible tissue extracts were 1.62 and 1.12 ng ml(-1) for ic-ELISA and TR-FIA detection, respectively. Two samples were spiked with QCA and analyzed by both methods. The recovery values ranged from 92.6% to 112.2% and the coefficients of variation were less than 15% for QCA spiking into swine tissue samples at concentrations of 2.5-50.0 μg kg(-1). Excellent correlations (r(2)=0.987-0.996) of the ic-ELISA/HPLC and TR-FIA/HPLC data were observed for processed samples. The results demonstrated that the ic-ELISA and TR-FIA methods were rapid and accurate for the residue detection of QCA in animal tissues. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. An innovative approach to sampling complex industrial emissions for use in animal toxicity tests: application to iron casting operations.

    PubMed

    Palmer, W G; Scholz, R C; Moorman, W J

    1983-03-01

    Sampling of complex mixtures of airborne contaminants for chronic animal toxicity tests often involves numerous sampling devices, requires extensive sampling time, and yields forms of collected materials unsuitable for administration to animals. A method is described which used a high volume, wet venturi scrubber for collection of respirable fractions of emissions from iron foundry casting operations. The construction and operation of the sampler are presented along with collection efficiency data and its application to the preparation of large quantities of samples to be administered to animals by intratracheal instillation.

  2. Detection of BVDV persistently infected animals in Belgium: evaluation of the strategy implemented.

    PubMed

    Letellier, C; De Meulemeester, L; Lomba, M; Mijten, E; Kerkhofs, P

    2005-11-15

    Until now, no official bovine virus diarrhea virus (BVDV) control program has been implemented in Belgium. The only legislation dealing with the detection of BVDV-infected animals concerns the purchase of animals. A strategy of control, based on the identification and elimination of persistently infected (PI) animals and the vaccination of cows before insemination has been designed in both the Northern and the Southern part of the country. The strategy of detection of PI animals relies on PCR testing of pools of blood. Individual blood samples corresponding to the positive pools are then tested by BVDV-antigen ELISA. A first evaluation of the measures already applied in Belgium is presented. Data obtained in 2003 are presented and discussed regarding the validation of the laboratory strategy, the prevalence of positive herds, the genotype of circulating viruses, the outcome of antigen positive animals and the need for improvement of the current legislation.

  3. Research and implementation of group animation based on normal cloud model

    NASA Astrophysics Data System (ADS)

    Li, Min; Wei, Bin; Peng, Bao

    2011-12-01

    Group Animation is a difficult technology problem which always has not been solved in computer Animation technology, All current methods have their limitations. This paper put forward a method: the Motion Coordinate and Motion Speed of true fish group was collected as sample data, reverse cloud generator was designed and run, expectation, entropy and super entropy are gotten. Which are quantitative value of qualitative concept. These parameters are used as basis, forward cloud generator was designed and run, Motion Coordinate and Motion Speed of two-dimensional fish group animation are produced, And two spirit state variable about fish group : the feeling of hunger, the feeling of fear are designed. Experiment is used to simulated the motion state of fish Group Animation which is affected by internal cause and external cause above, The experiment shows that the Group Animation which is designed by this method has strong Realistic.

  4. [Occurrence of Giardia species and genotypes in humans and animals in Wielkopolska region, Poland].

    PubMed

    Solarczyk, Piotr

    2009-01-01

    Giardia is the most common intestinal protozoan parasite found in humans and animals worldwide. Although it has been known for three hundred years, the nomenclature, taxonomy, host specificity, and pathogenicity of Giardia still arouse numerous controversies and ambiguities. Giardia is classified into six species, that are characterised by various ranges of hosts. The most dubious species is G. intestinalis, which includes a dozen or so genotypes, and only two of them (genotype A and B) have wide ranges of hosts, including humans. Moreover, in some genotype assemblages of G. intestinalis certain subgenotypes were distinguished and it was proven that in the same host species various subgenotypes of this parasite may occur. Bearing in mind the significant genetic heterogeneity of G. intestinalis and the fact that various genotypes and subgenotypes of this parasite are characterised by the broad or narrow host specificity, the data concerning the frequency of giardiosis occurrence are insufficient. It is necessary to use molecular biology techniques in order to define the genotype and/or the subgenotype of G. intestinalis that are found in humans and in certain animal species. Furthermore, since more and more pieces of evidence connected with a possibility of the sexual recombination of Giardia are gathered, it is unknown if genotypes and subgenotypes of this parasite are stable in time. The aim of this thesis was to define the frequency of Giardia occurrence in humans and animals in Wielkopolska region, to identify species and genotypes of Giardia that occur in humans and animals, as well as to obtain an axenic culture of the chosen isolates of Giardia from animals and to compare the sequence of the beta-giardin gene fragment obtained from the DNA isolated from cysts and trophozoites in order to check if the axenisation of G. intestinalis leads to the selection of genotypes or if Giardia genotypes are stable in time. Altogether, 2183 faecal samples were examined for the presence of Giardia cysts; 447 faecal samples were taken from 232 persons coming from 5 cities situated in Wielkopolska, and 1736 faecal samples were obtained from 123 animal species, including: 266 faecal samples from 113 species of animals kept in the Zoological Garden in Poznań, 1286 samples from 4 species of breeding animals, 118 samples from dogs, and 66 samples from 5 species of wild animals. Faecal samples were taken from animals coming from 25 places in Wielkopolska. Moreover, seven isolates of G. intestinalis were used in the studies, which were obtained from various species of hosts and kept in an axenic in vitro culture. Microscopic, molecular and bio-informative methods were used in the studies. From each faecal sample fresh smears were made in a 0.6% solution of physiological salt and in Lugol's solution, as well as a permanent smear stained with trichrome was made. Moreover, the following molecular techniques were implemented in the studies: DNA extraction and purification, the PCR technique (two molecular markers), electrophoresis and visualisation of PCR products, and sequencing. A fragment of the beta-giardin gene was used as a molecular marker in order to define the genotype and subgenotype of Giardia. Only in the case of genotyping of two Giardia isolates obtained from Peromyscus eremicus another molecular marker (SSU rRNA)was additionally used. Some widely available computer programmes (Chromas, CAP 3, BioEdit, BLASTn, MEGA version 4.0) were utilised in the analysis of the sequence of the beta-giardin gene fragment and in the phylogenetic analysis. The culture of Giardia trophozoites was established to compare the sequence of the partial beta-giardin gene from cysts and trophozoites. Concentration and purification of Giardia cysts in the saccharose gradient, and the excystation technique were applied in the studies to obtaining an axenic in vitro culture. In this study, Giardia cysts were found in 12 faecal samples obtained from 3 persons and 5 animal species. Giardia cysts were found only in faecal samples from humans living in Poznań and the samples obtained from animals coming from Poznań and around Puszczykowo. The highest frequency of infection was stated in domestic animals (2.5%) and in animals kept in the Zoological Garden (2.0%), whereas a slightly lower frequency was noticed in wild animals (1.5%) and in humans (1.3%). No Giardia cysts were found in the faecal samples collected from breeding animals. Two new species of Giardia hosts were identified, namely Rhinella marina and Peromyscus eremicus; however, due to a minimal amount of faecal samples supplied for the study it was impossible to define the species and genotype of this parasite. PCR products (the partial of beta-giardin gene) were obtained in seven faecal samples out of the ten studied, including three samples from people and four faecal samples derived from three animal species (i.e. dog, tamandua, red deer). Moreover, molecular characterization of seven Giardia isolates from three persons and four animal species (red-bellied monkey, silver marmoset, Thomson's gazelle, and sheep) kept in an axenic in vitro culture was performed. Based on the beta-giardin sequence fragment analysis, four assemblages of G. intestinalis genotypes were identified (A, B, C and D). In humans, A and B G. intestinalis genotypes and three subgenotypes, including a cosmopolitan subgenotype A2 and two new subgenotypes A and B were detected. Furthermore, four G. intestinalis genotypes were found in animals, including three genotypes which are non-infectious to humans, namely: genotypes C and D in dogs and a cervids-specific genotype A in red deer (Cervus elaphus), which indicate that these animals do not constitute the source of infection to humans. On the other hand, in a tamandua from the Zoological Garden in Poznań a new subgenotype B of G. intestinalis was identified, which due to a close relationship with Giardia isolates obtained from humans is potentially infectious to man. In none of the studied faecal samples a mixed infection of Giardia was found. To date, nine sequences of the partial beta-giardin gene have been deposited in the National Center for Biotechnology Information (NCBI), including two sequences of Giardia isolates obtained from humans (GenBank accession numbers FJ009207, FJ009208), three sequences of isolate obtained from red deer (GenBank accession numbers EU621373, EU626198, EU216429), two sequences of both Giardia isolates obtained from dogs (GenBank accession numbers FJ009205, FJ009206), and the single sequences obtained from tamandua (GenBank accession number FJ009209) and from Thomson's gazelle (GenBank accession number EU626199). According to the literature, an axenic in vitro culture of G. intestinalis was obtained from a red deer for the first time. Based on the analysis of the sequence of the beta-giardin gene fragment obtained from the DNA isolated from cysts and trophozoites it was proven that the red deer was infected with a single population of Giardia and that during the axenisation of the culture no mutation in the DNA of the parasite's trophozoites took place. Probably the time distance that the DNA was isolated from the trophozoites kept in the culture was too short to cause the mutation. This suggestion is confirmed by the results of the genotyping of seven G. intestinalis isolates obtained from various host species and kept in an axenic in vitro culture for at least a number of years. Based on the molecular characteristics it was stated that all the studied isolates from the axenic culture were identical and belonged to the same assemblage B. The comparision with the sequences from GenBank database revealed that all mentioned isolates were 99% similar to the sequence of Giardia Nij5 isolate obtained from a person from the Netherlands and characterised as genotype B1. Due to the sameness of the molecular marker sequences it seems improbable that the identical G. intestinalis genotype occurred in various time periods (the largest difference was 14 years) in humans and in a number of animal species in diverse areas of Wielkopolska region. Quite opposite, the long-term keeping of these isolates in the homogenous conditions of an axenic in vitro culture leads to the selection of a genotype or proves the instability of genotypes of this parasite. Long-term studies need to be conducted in order to verify these hypothesis. Their results will have a key meaning in explaining the genetic structure of the Giardia population and in understanding the molecular epidemiology of giardiosis.

  5. Hyperspectral imaging of endogenous fluorescent metabolic molecules to identify pain states in central nervous system tissue

    NASA Astrophysics Data System (ADS)

    Staikopoulos, Vasiliki; Gosnell, Martin E.; Anwer, Ayad G.; Mustafa, Sanam; Hutchinson, Mark R.; Goldys, Ewa M.

    2016-12-01

    Fluorescence-based bio-imaging methods have been extensively used to identify molecular changes occurring in biological samples in various pathological adaptations. Auto-fluorescence generated by endogenous fluorescent molecules within these samples can interfere with signal to background noise making positive antibody based fluorescent staining difficult to resolve. Hyperspectral imaging uses spectral and spatial imaging information for target detection and classification, and can be used to resolve changes in endogenous fluorescent molecules such as flavins, bound and free NADH and retinoids that are involved in cell metabolism. Hyperspectral auto-fluorescence imaging of spinal cord slices was used in this study to detect metabolic differences within pain processing regions of non-pain versus sciatic chronic constriction injury (CCI) animals, an established animal model of peripheral neuropathy. By using an endogenous source of contrast, subtle metabolic variations were detected between tissue samples, making it possible to distinguish between animals from non-injured and injured groups. Tissue maps of native fluorophores, flavins, bound and free NADH and retinoids unveiled subtle metabolic signatures and helped uncover significant tissue regions with compromised mitochondrial function. Taken together, our results demonstrate that hyperspectral imaging provides a new non-invasive method to investigate central changes of peripheral neuropathic injury and other neurodegenerative disease models, and paves the way for novel cellular characterisation in health, disease and during treatment, with proper account of intrinsic cellular heterogeneity.

  6. Simultaneous measurement of NAD metabolome in aged mice tissue using liquid chromatography tandem-mass spectrometry.

    PubMed

    Yaku, Keisuke; Okabe, Keisuke; Nakagawa, Takashi

    2018-06-01

    Nicotinamide adenine dinucleotide (NAD) is a major co-factor that mediates multiple biological processes including redox reaction and gene expression. Recently, NAD metabolism has received considerable attention because administration of NAD precursors exhibited beneficial effects against aging-related metabolic disorders in animals. Although numerous studies have reported that NAD levels decline with aging in multiple animal tissues, the pathway and kinetics of NAD metabolism in aged organs are not completely understood. To determine the NAD metabolism upon aging, we developed targeted metabolomics based on an LC/MS/MS system. Our method is simple and applicable to crude biological samples, including culture cells and animal tissues. Unlike a conventional enzymatic cycling assay, our approach can determine NAD and NADH (reduced form of NAD) by performing a single sample preparation. Further, we validated our method using biological samples and investigated the alteration of the NAD metabolome during aging. Consistent with previous reports, the NAD levels in the liver and skeletal muscle decreased with aging. Further, we detected a significant increase in nicotinamide mononucleotide and nicotinamide riboside in the kidney upon aging. The LC/MS/MS-based NAD metabolomics that we have developed is extensively applicable to biomedical studies, and the results will present innovative ideas for the aging studies, especially for that of NAD metabolism. Copyright © 2018 John Wiley & Sons, Ltd.

  7. The rumen microbiome as a reservoir of antimicrobial resistance and pathogenicity genes is directly affected by diet in beef cattle.

    PubMed

    Auffret, Marc D; Dewhurst, Richard J; Duthie, Carol-Anne; Rooke, John A; John Wallace, R; Freeman, Tom C; Stewart, Robert; Watson, Mick; Roehe, Rainer

    2017-12-11

    The emergence and spread of antimicrobial resistance is the most urgent current threat to human and animal health. An improved understanding of the abundance of antimicrobial resistance genes and genes associated with microbial colonisation and pathogenicity in the animal gut will have a major role in reducing the contribution of animal production to this problem. Here, the influence of diet on the ruminal resistome and abundance of pathogenicity genes was assessed in ruminal digesta samples taken from 50 antibiotic-free beef cattle, comprising four cattle breeds receiving two diets containing different proportions of concentrate. Two hundred and four genes associated with antimicrobial resistance (AMR), colonisation, communication or pathogenicity functions were identified from 4966 metagenomic genes using KEGG identification. Both the diversity and abundance of these genes were higher in concentrate-fed animals. Chloramphenicol and microcin resistance genes were dominant in samples from forage-fed animals (P < 0.001), while aminoglycoside and streptomycin resistances were enriched in concentrate-fed animals. The concentrate-based diet also increased the relative abundance of Proteobacteria, which includes many animal and zoonotic pathogens. A high ratio of Proteobacteria to (Firmicutes + Bacteroidetes) was confirmed as a good indicator for rumen dysbiosis, with eight cases all from concentrate-fed animals. Finally, network analysis demonstrated that the resistance/pathogenicity genes are potentially useful as biomarkers for health risk assessment of the ruminal microbiome. Diet has important effects on the complement of AMR genes in the rumen microbial community, with potential implications for human and animal health.

  8. Replacing animal use in physiology and pharmacology teaching in selected universities in Eastern Europe--charting a way forward.

    PubMed

    Dewhurst, David G; Kojic, Zvezdana Z

    2011-03-01

    The aims of this study were to explore the use of animals in teaching and the implementation of innovative technology-based teaching practices across a small sample of universities in Eastern Europe. The research methods used were a questionnaire circulated four weeks before a workshop took place (in October 2009, in Belgrade, Serbia), as well as focused, face-to-face group discussions, led by one of the authors during the workshop. Twenty-two faculty (physiologists and pharmacologists), from 13 Eastern European countries, attended the meeting. Fourteen of the eighteen schools represented at the workshop were making use of animals, in some instances in quite large numbers, for their teaching. For example, a single department at a Romanian university used over 250 animals per annum, and at least 1130 animals were used, per annum, across all of the institutions. The species used in largest numbers were the rat (34%), frog/toad (29%), mouse (22%), rabbit (10%), guinea-pig (4%) and dog (1%). None of the universities sampled had implemented institution-wide virtual learning environments (VLEs), although there were isolated instances of local use of VLEs. There was relatively little current use of technology-based teaching and learning resources, but there was considerable enthusiasm to modernise teaching and to introduce innovative learning and teaching methods. The major perceived barrier to the introduction of replacement alternatives was the lack of versions in local languages. There was a consensus view that developing local language exemplars and evaluating their usefulness was likely to have the greatest impact on animal use, at least in the short-term. 2011 FRAME.

  9. DNA-based approach to aging martens (Martes americana and M. caurina)

    Treesearch

    Jonathan N. Pauli; John P. Whiteman; Bruce G. Marcot; Terry M. McClean; Merav Ben-David

    2011-01-01

    Demographic structure is central to understanding the dynamics of animal populations. However, determining the age of free-ranging mammals is difficult, and currently impossible when sampling with noninvasive, genetic-based approaches. We present a method to estimate age class by combining measures of telomere lengths with other biologically meaningful covariates in a...

  10. Detection of Leptospiral DNA in the Urine of Donkeys on the Caribbean Island of Saint Kitts

    PubMed Central

    Grevemeyer, Bernard; Vandenplas, Michel; Beigel, Brittney; Cho, Ellen; Willingham, Arve Lee; Verma, Ashutosh

    2017-01-01

    Leptospirosis is a global zoonosis caused by pathogenic spirochetes classified within the genus Leptospira. Leptospires live in the proximal renal tubules of reservoir or chronic carrier animals, and are shed in the urine. Naïve animals acquire infection either when they come in direct contact with a reservoir or infected animals or by exposure to environmental surface water or soil that is contaminated with their urine. In this study, urine samples from a herd of donkeys on the Caribbean island of St. Kitts were screened using a TaqMan-based real-time quantitative polymerase chain reaction (qPCR) targeting a pathogen-specific leptospiral gene, lipl32. Out of 124 clinically normal donkeys, 22 (18%) tested positive for leptospiral DNA in their urine. Water samples from two water troughs used by the donkeys were also tested, but were found to be free from leptospiral contamination. Detection of leptospiral DNA in the urine of clinically healthy donkeys may point to a role that these animals play in the maintenance of the bacteria on St. Kitts. PMID:29056661

  11. Use of DNA from bite marks can determine species and individual animals that attack humans

    USGS Publications Warehouse

    Farley, Sean; Talbot, Sandra L.; Sage, George K.; Sinnott, Rick; Coltrane, Jessica

    2014-01-01

    During the summer of 2008, 6 documented attacks and close encounters with brown bears (Ursus arctos) occurred in the greater Anchorage, Alaska (USA) area. We discuss findings from 2 incidents in which people were mauled within 2 km of each other over a 6-week period and in which it was assumed that a single animal was responsible. To ensure public safety, authorities killed a brown bear implicated in the attacks by circumstantial evidence, though it was not known a priori that the animal was responsible. We extracted DNA from hairs and bite sites on the clothing of both victims and determined species and individual identity of the animal(s) involved in both incidents. Genetic data indicated the brown bear killed by authorities was responsible for one of the maulings, but not both. This research demonstrates that DNA-based techniques, with appropriate sampling, can provide unambiguous identification of animals involved in attacks, as well as provide reasonable justification for excluding others. Because DNA-based techniques can unequivocally identify individual bears carrying out attacks, they should be considered a standard method employed in wildlife attack investigations.

  12. COMADRE: a global data base of animal demography.

    PubMed

    Salguero-Gómez, Roberto; Jones, Owen R; Archer, C Ruth; Bein, Christoph; de Buhr, Hendrik; Farack, Claudia; Gottschalk, Fränce; Hartmann, Alexander; Henning, Anne; Hoppe, Gabriel; Römer, Gesa; Ruoff, Tara; Sommer, Veronika; Wille, Julia; Voigt, Jakob; Zeh, Stefan; Vieregg, Dirk; Buckley, Yvonne M; Che-Castaldo, Judy; Hodgson, David; Scheuerlein, Alexander; Caswell, Hal; Vaupel, James W

    2016-03-01

    The open-data scientific philosophy is being widely adopted and proving to promote considerable progress in ecology and evolution. Open-data global data bases now exist on animal migration, species distribution, conservation status, etc. However, a gap exists for data on population dynamics spanning the rich diversity of the animal kingdom world-wide. This information is fundamental to our understanding of the conditions that have shaped variation in animal life histories and their relationships with the environment, as well as the determinants of invasion and extinction. Matrix population models (MPMs) are among the most widely used demographic tools by animal ecologists. MPMs project population dynamics based on the reproduction, survival and development of individuals in a population over their life cycle. The outputs from MPMs have direct biological interpretations, facilitating comparisons among animal species as different as Caenorhabditis elegans, Loxodonta africana and Homo sapiens. Thousands of animal demographic records exist in the form of MPMs, but they are dispersed throughout the literature, rendering comparative analyses difficult. Here, we introduce the COMADRE Animal Matrix Database, an open-data online repository, which in its version 1.0.0 contains data on 345 species world-wide, from 402 studies with a total of 1625 population projection matrices. COMADRE also contains ancillary information (e.g. ecoregion, taxonomy, biogeography, etc.) that facilitates interpretation of the numerous demographic metrics that can be derived from its MPMs. We provide R code to some of these examples. We introduce the COMADRE Animal Matrix Database, a resource for animal demography. Its open-data nature, together with its ancillary information, will facilitate comparative analysis, as will the growing availability of databases focusing on other aspects of the rich animal diversity, and tools to query and combine them. Through future frequent updates of COMADRE, and its integration with other online resources, we encourage animal ecologists to tackle global ecological and evolutionary questions with unprecedented sample size. © 2016 The Authors. Journal of Animal Ecology published by John Wiley & Sons Ltd on behalf of British Ecological Society.

  13. A capture-recapture survival analysis model for radio-tagged animals

    USGS Publications Warehouse

    Pollock, K.H.; Bunck, C.M.; Winterstein, S.R.; Chen, C.-L.; North, P.M.; Nichols, J.D.

    1995-01-01

    In recent years, survival analysis of radio-tagged animals has developed using methods based on the Kaplan-Meier method used in medical and engineering applications (Pollock et al., 1989a,b). An important assumption of this approach is that all tagged animals with a functioning radio can be relocated at each sampling time with probability 1. This assumption may not always be reasonable in practice. In this paper, we show how a general capture-recapture model can be derived which allows for some probability (less than one) for animals to be relocated. This model is not simply a Jolly-Seber model because it is possible to relocate both dead and live animals, unlike when traditional tagging is used. The model can also be viewed as a generalization of the Kaplan-Meier procedure, thus linking the Jolly-Seber and Kaplan-Meier approaches to survival estimation. We present maximum likelihood estimators and discuss testing between submodels. We also discuss model assumptions and their validity in practice. An example is presented based on canvasback data collected by G. M. Haramis of Patuxent Wildlife Research Center, Laurel, Maryland, USA.

  14. An analysis of wilderness water in Kings Canyon, Sequoia, and Yosemite national parks for coliform and pathologic bacteria.

    PubMed

    Derlet, Robert W; Carlson, James R

    2004-01-01

    To determine the prevalence of coliform and potentially pathogenic bacteria in remote backcountry alpine lakes and streams of national parks in the Sierra Nevada mountains. Water was sampled at 55 predetermined lakes and streams that would stratify the risk, based on sites used by backpackers, sites used by pack animals, and uncontaminated wild areas. Sites were distributed among Kings Canyon (15), Sequoia (17), and Yosemite (23). Water was collected using Millipore bacterial samplers, which provided specific counts of coliform and other bacteria in each water sample and also served as a transport media from the wilderness to the laboratory. On return to the laboratory, bacteria were harvested from the samplers and subjected to specific identification and qualitative analysis using standard microbiology techniques for the analysis of water. Coliform bacteria were detected in 22 of the 55 sites. All of these sites were below areas used by backpackers or pack animals. Thirty-three sites were free of coliforms. These sites included both those used lightly by backpackers and those with no human or domestic animal use. All samples contained expected amounts of normal aquatic bacteria including Pseudomonas, Rahnella aquatilis, Serratia spp, and nonpathogenic species of Yersinia. Most sampling sites in these national parks are free of coliform or pathogenic organisms. Low levels of coliform bacteria are found in some bodies of water where the watershed has been affected by human or pack animal travel.

  15. Animation of 'Dodo' and 'Goldilocks' Trenches

    NASA Technical Reports Server (NTRS)

    2008-01-01

    [figure removed for brevity, see original site] Click on image for animation

    A pan and zoom animation of the informally named 'Dodo' (on left) and 'Goldilocks' (on right) trenches as seen by the Surface Stereo Imager (SSI) aboard NASA's Phoenix Mars Lander. This animation was based on conditions on the Martian surface on Sol 17 (June 11, 2008), the 17th Martian day of the mission. 'Baby Bear' is the name of the sample taken from 'Goldilocks' and delivered to the Thermal and Evolved-Gas Analyzer (TEGA) instrument.

    The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  16. Detection of Rickettsia helvetica in Ixodes ricinus infesting wild and domestic animals and in a botfly larva (Cephenemyia stimulator) infesting roe deer in Germany.

    PubMed

    Scheid, Patrick; Speck, Stephanie; Schwarzenberger, Rafael; Litzinger, Mark; Balczun, Carsten; Dobler, Gerhard

    2016-10-01

    Ixodes ricinus is a well-known vector of different human pathogens including Rickettsia helvetica. The role of wild mammals in the distribution and probable maintenance of Rickettsia in nature is still to be determined. We therefore investigated various parasites from different wild mammals as well as companion animals for the presence of Rickettsia. A total of 606 I. ricinus, 38 Cephenemyia stimulator (botfly larvae), one Dermacentor reticulatus, 24 Haematopinus suis (hog lice) and 30 Lipoptena cervi (deer flies) were collected from free-ranging animals during seasonal hunting, and from companion animals. Sample sites included hunting leases at three main sampling areas and five additional areas in West and Central Germany. All collected parasites were screened for Rickettsia spp. and I. ricinus were investigated for tick-borne encephalitis virus (TBEV) in addition. While no TBEV was detected, the minimum infection rate (MIR) of I. ricinus with Rickettsia was 4.1% referring to all sampling sites and up to 6.9% at the main sampling site in Koblenz area. Sequencing of a fragment of the ompB gene identified R. helvetica. Approximately one third (29.5%) of the animals carried Rickettsia-positive ticks and the MIR in ticks infesting wild mammals ranged from 4.1% (roe deer) to 9.5%. These data affirm the widespread distribution of R. helvetica in Germany. One botfly larva from roe deer also harboured R. helvetica. Botfly larvae are obligate parasites of the nasal cavity, pharynx and throat of cervids and feed on cell fragments and blood. Based on this one might hypothesise that R. helvetica likely induces rickettsemia in cervids thus possibly contributing to maintenance and distribution of this rickettsia in the field. Copyright © 2016 Elsevier GmbH. All rights reserved.

  17. Development of a milk and serum ELISA test for the detection of Teladorsagia circumcincta antibodies in goats using experimentally and naturally infected animals.

    PubMed

    Malama, Eleni; Hoffmann-Köhler, Peggy; Biedermann, Insa; Koopmann, Regine; Krücken, Jürgen; Molina, José Manuel; Moreno, Alvaro Martinez; von Samson-Himmelstjerna, Georg; Sotiraki, Smaragda; Demeler, Janina

    2014-10-01

    Teladorsagia circumcincta is among the most important gastrointestinal parasites in small ruminants and the predominant species in Southern European goats. Parasite control is largely based on metaphylactic/preventative treatments, which is often seen as non-sustainable anymore. The reasons are increased consumer demand to reduce chemicals in livestock production and anthelmintic resistance against the common drugs. This study aimed at the development of a T. circumcincta-enzyme-linked immunosorbent assay (ELISA) specifically for goats. Samples were obtained from goats raised parasite-free or infected experimentally. Sampling continued during the following pasture season and housing period. The sensitivity for the use in bulk milk samples as an indicator of T. circumcincta infection levels in grazing goats was examined. The ELISA enables clear differentiation of negative and positive animals. With a specificity of 100% negative cut-off values for serum and milk were 0.294 and 0.228 (sensitivity, 95%). Positive cut-off values (sensitivity, 90%) were 0.606 (serum) and 0.419 (milk), while a sensitivity of 95% resulted in 0.509 and 0.363, respectively. The grey-zone between negative/positive cut-offs was introduced to deal with animals in pre-patency and decreasing antibody levels after infection. There was no cross reactivity for Trichostrongylus colubriformis and Cooperia oncophora while for Haemonchus contortus and Fasciola hepatica it cannot be fully excluded currently. In bulk milk samples, 5% of the milk had to be contributed from animals infected with T. circumcincta to be detected as positive. The results derived from experimentally and naturally infected as well as parasite naïve animals indicate the potential of the ELISA to be used in targeted anthelmintic treatment regimes in goats.

  18. Identification of Mycobacterium bovis in bovine clinical samples by PCR species-specific primers.

    PubMed Central

    Romero, R E; Garzón, D L; Mejía, G A; Monroy, W; Patarroyo, M E; Murillo, L A

    1999-01-01

    Tuberculosis, caused by Mycobacterium bovis is emerging as the most important disease affecting cattle. Furthermore, it results in a major public health problem when transmitted to humans. Due to its difficult and non-specific diagnosis, M. bovis has been declared to be one of the etiologic agents causing significant economic loss in the cattle industry. Our group evaluated a more rapid and specific method, based on a new polymerase chain reaction species-specific primers, which amplifies a 470-base pair fragment of the M. bovis genome. A total of 275 milk-producing cows were studied by intradermal tuberculin test (ITT) which gave 184 positive and 91 negative cases. From them, 50 animals were taken from a cattle ranch free of tuberculosis. Three different samples were collected from each animal (blood, nasal mucus, and milk). Positive results were obtained from 26 animals by PCR (11.4%), 1 by bacteriological culturing (0.4%) and 1 by bacilloscopy (0.4%). This finding suggests, as in previous reports, that ITT, normally used for bovine tuberculosis detection, has the inconvenience of having a broad range of specificity and sensitivity, and the PCR technique is a more specific and sensitive test to detect infection associated with M. bovis. Therefore, we propose this PCR assay as a useful tool in the epidemiological characterization of infected animals in areas considered to be at high risk of transmission. Images Figure 1. PMID:10369566

  19. Development of a polymerase chain reaction assay for the detection of pseudorabies virus in clinical samples

    PubMed Central

    Pérez, Lester J.; de Arce, Heidy Díaz

    2009-01-01

    Aujeszky’s disease, also known as pseudorabies causes severe economic losses in swine industry and affects the pig husbandry all over the world. The conventional diagnostic procedure is time-consuming and false-negative results may occur in submissions from latently infected animals. The development, optimization and evaluation of a polymerase chain reaction (PCR) assay are presented for the diagnosis of pseudorabies infection. This assay was based on the amplification of a highly conserved viral gD gene fragment. PCR products of the expected size were obtained from PRV strains. Non-specific reactions were not observed when a related herpesvirus, other porcine DNA genome viruses and uninfected cells were used to assess PCR. The analytical sensitivity of the test was estimated to be 1.34 TCID50/ 50 uL. The analysis of tissue homogenate samples from naturally infected animals proved the potential usefulness of the method for a rapid disease diagnosis from field cases. A rapid, sensitive and specific PCR-based diagnostic assay to detect pseudorabies virus in clinical samples is provided. PMID:24031383

  20. Buccal Swabbing as a Noninvasive Method To Determine Bacterial, Archaeal, and Eukaryotic Microbial Community Structures in the Rumen

    PubMed Central

    Kirk, Michelle R.; Jonker, Arjan; McCulloch, Alan

    2015-01-01

    Analysis of rumen microbial community structure based on small-subunit rRNA marker genes in metagenomic DNA samples provides important insights into the dominant taxa present in the rumen and allows assessment of community differences between individuals or in response to treatments applied to ruminants. However, natural animal-to-animal variation in rumen microbial community composition can limit the power of a study considerably, especially when only subtle differences are expected between treatment groups. Thus, trials with large numbers of animals may be necessary to overcome this variation. Because ruminants pass large amounts of rumen material to their oral cavities when they chew their cud, oral samples may contain good representations of the rumen microbiota and be useful in lieu of rumen samples to study rumen microbial communities. We compared bacterial, archaeal, and eukaryotic community structures in DNAs extracted from buccal swabs to those in DNAs from samples collected directly from the rumen by use of a stomach tube for sheep on four different diets. After bioinformatic depletion of potential oral taxa from libraries of samples collected via buccal swabs, bacterial communities showed significant clustering by diet (R = 0.37; analysis of similarity [ANOSIM]) rather than by sampling method (R = 0.07). Archaeal, ciliate protozoal, and anaerobic fungal communities also showed significant clustering by diet rather than by sampling method, even without adjustment for potentially orally associated microorganisms. These findings indicate that buccal swabs may in future allow quick and noninvasive sampling for analysis of rumen microbial communities in large numbers of ruminants. PMID:26276109

  1. Antibody responses in New World camelids with tuberculosis caused by Mycobacterium microti.

    PubMed

    Lyashchenko, K P; Greenwald, R; Esfandiari, J; Meylan, M; Burri, I Hengrave; Zanolari, P

    2007-12-15

    Antibody responses in New World camelids (NWC) infected with Mycobacterium microti were studied by two serological methods, multiantigen print immunoassay (MAPIA) and lateral-flow-based rapid test (RT). Serum samples were collected during 2004-2006 from 87 animals including 1 alpaca and 7 llamas with confirmed or suspected M. microti infection, 33 potentially exposed but clinically healthy animals from known infected herds, and 46 control NWC from herds where infection had not been previously diagnosed. The serological assays correctly identified infection status in 97% (MAPIA) or 87% (RT) cases. In three llamas with confirmed M. microti infection and one llama with gross pathology suggestive of disease, for which multiple serum samples collected over time were available, the antibody-based tests showed positive results 1-2 years prior to the onset of clinical signs or being found dead. In MAPIA, MPB83 protein was identified to be an immunodominant serological target antigen recognized in NWC infected with M. microti. With the limited number of animals tested in this study, the serological assays demonstrated the potential for convenient, rapid, and accurate diagnosis of M. microti infection in live llamas and alpacas.

  2. Verbal fluency in elderly with and without hypertension and diabetes from the FIBRA study in Ermelino Matarazzo

    PubMed Central

    Morelli, Nathalia Lais; Cachioni, Meire; Lopes, Andrea; Batistoni, Samila Sathler Tavares; Falcão, Deusivania Vieira da Silva; Neri, Anita Liberalesso; Yassuda, Monica Sanches

    2017-01-01

    ABSTRACT. Background: There are few studies on the qualitative variables derived from the animal category verbal fluency test (VF), especially with data originating from low-income samples of community-based studies. Objective: To compare elderly with and without hypertension (HTN) and diabetes mellitus (DM) regarding the total number of animals spoken, number of categories, groups and category switches on the VF test. Methods: We used the database of the FIBRA (Frailty in Brazilian Elderly) community-based study. The variables number of Categories, Groups and Category Switches were created for each participant. The total sample (n = 384) was divided into groups of elderly who reported having HTN, DM, both HTN and DM, or neither of these conditions. Results: There were no significant differences between the groups with and without these chronic diseases for VF total score or for the qualitative variables. Conclusion: Among independent community-dwelling elderly, the qualitative variables derived from the VF animal category may not add information regarding the cognitive profile of elderly with chronic diseases. Total VF score and the qualitative variables Category, Group and Switching did not differentiate elderly with and without HTN and DM. PMID:29354222

  3. Kind News Mini Edition, Jrs.

    ERIC Educational Resources Information Center

    Children & Animals, 1987

    1987-01-01

    Contains suggestions for students regarding making up nicknames for animals based on their observations. Also includes information about the overpopulation of pets and urges students to write letters to their newspaper editors about this problem. A sample letter is provided. (TW)

  4. The effect of inmates' self-reported childhood and adolescent animal cruelty: motivations on the number of convictions for adult violent interpersonal crimes.

    PubMed

    Hensley, Christopher; Tallichet, Suzanne E

    2008-04-01

    Few researchers have investigated the potentially predictive power of motives for childhood and adolescent animal cruelty as it is associated with interpersonal violence in adulthood. Based on a sample of 261 inmates at medium- and maximum-security prisons in a southern state, the present study examines the relationship among several retrospectively reported motives (anger, fun, dislike, and imitation) for animal cruelty and violent crime convictions (assault, rape, and murder). Almost half reported abusing animals out of anger, whereas more than one third did so for fun. Dislike for the animal and imitation were less frequently occurring motives. Participants who abused animals at an earlier age and those who did so out of anger or for fun were more likely to repeat the offense. Regression analyses revealed that abusing an animal out of fun in their youth was the most statistically salient motive for predicting later interpersonal violence as adults.

  5. Bovine trypanosomosis in the Bolivian Pantanal.

    PubMed

    Gonzales, J L; Chacon, E; Miranda, M; Loza, A; Siles, L M

    2007-05-15

    Trypanosomosis caused by Trypanosoma vivax has been a constraint for cattle production in the Bolivian lowlands, since it was introduced in 1996. Flooded areas like the Bolivian Pantanal have a suitable environment for the presence and transmission of Salivarian trypanosomes and farmers from that region often report trypanosomosis-like problems on their farms. The objective of the present study, therefore, was to characterize the epidemiology of bovine trypanosomosis in the Bolivian Pantanal. In order to achieve this objective, 202 cattle from the province of Angel Sandoval and 209 cattle from the province of German Busch were randomly sampled (the Pantanal is located in both provinces). Twenty-nine farms in both provinces were visited, the farmers interviewed, and biologic samples collected from their cattle. Samples were submitted for parasitological and PCR evaluation and the prevalence of bovine trypanosomosis was estimated for each province. Laboratory results were correlated with the sampled animals packed cell volume (PCV) and body condition (BC) scores and the observed T. vivax parasites measured for morphometry analysis. Results from this study show differences in morphometric measures between T. vivax parasites from each province. Differences between provinces were also observed in the T. vivax-related disease situation. While in Angel Sandoval the PCV and BC of T. vivax-affected animals were significantly lower than those of the T. vivax-negative animals, in German Busch no differences were observed in the PCV and BC of T. vivax-positive or negative animals. Animal prevalence of T. vivax in Angel Sandoval was 27.79% (95% CI: 14.52-44.28) and in German Busch was 19.03% (95% CI: 9.19-30.75). The T. evansi animal prevalence in each province was 0.99% (95% CI: 0.27-2.99) and 5.71% (95% CI: 2.43-12.19), respectively. Based on questionnaire and laboratory results, it was concluded that trypanosomosis is a primary constraint for cattle production in the Bolivian Pantanal.

  6. Isotopic change in the tissues of Bothrops atrox in captivity collected from environments of the eastern Amazon

    NASA Astrophysics Data System (ADS)

    Martinez, M. G.; Chalkidis, H. D.; Amazonas, D. R.; da Silva, A. M.; De Oliveira, R., Jr.; Camargo, P. B.

    2013-12-01

    The Bothrops atrox is little studied because it is sympatric Amazonian animals, and very little is known about the ecology and natural history of this species. It has a generalist diet and the distribution of this species is very wide. The adult animals forage mostly on the ground, while the younger animals prefer to stay on the vegetation. They are easily find in the rainy months in areas near lakes and seasonally flooded and are difficult to find in the driest months, a period where there is less availability of preys in these environments. Due to its aggressiveness, is considered one of the most feared snakes in South America and in the eastern Amazon, being responsible for the largest number of snakebites in the region. Through stable isotope carbon-13 and nitrogen-15, is intended to characterize the variations of the feeding habits of these collected animals in different environments and also when they are kept in captivity, feeding the animal's bioterium. The serpents were collected in environments with different land uses, such as native forest, savannah, pasture and have been brought to the serpentarium Integrated College Tapajos (FIT), being retained in order to Samplings throughout the experiment with feeding mice's own bioterium. When these snakes came from different locations, samples were collected scales and blood (T0), before receiving the new supply (captive), and every time we fed the mice the vivarium, new tissue samples were collected, (T1, T2, T3) to exchange all the nature of food for the food captivity.Based on the results of δ13C and δ15N, the samples collected in the tissues of snakes of different environments (nature and captivity), it was observed that changes in food sources reflect changes in tissues (blood and scales), also reflecting the production of poison different periods of turnover of absorbed material in those tissues, contributing to the study of animal ecology and behavior in relation to habitat.

  7. Assessing host-specificity of Escherichia coli using a supervised learning logic-regression-based analysis of single nucleotide polymorphisms in intergenic regions.

    PubMed

    Zhi, Shuai; Li, Qiaozhi; Yasui, Yutaka; Edge, Thomas; Topp, Edward; Neumann, Norman F

    2015-11-01

    Host specificity in E. coli is widely debated. Herein, we used supervised learning logic-regression-based analysis of intergenic DNA sequence variability in E. coli in an attempt to identify single nucleotide polymorphism (SNP) biomarkers of E. coli that are associated with natural selection and evolution toward host specificity. Seven-hundred and eighty strains of E. coli were isolated from 15 different animal hosts. We utilized logic regression for analyzing DNA sequence data of three intergenic regions (flanked by the genes uspC-flhDC, csgBAC-csgDEFG, and asnS-ompF) to identify genetic biomarkers that could potentially discriminate E. coli based on host sources. Across 15 different animal hosts, logic regression successfully discriminated E. coli based on animal host source with relatively high specificity (i.e., among the samples of the non-target animal host, the proportion that correctly did not have the host-specific marker pattern) and sensitivity (i.e., among the samples from a given animal host, the proportion that correctly had the host-specific marker pattern), even after fivefold cross validation. Permutation tests confirmed that for most animals, host specific intergenic biomarkers identified by logic regression in E. coli were significantly associated with animal host source. The highest level of biomarker sensitivity was observed in deer isolates, with 82% of all deer E. coli isolates displaying a unique SNP pattern that was 98% specific to deer. Fifty-three percent of human isolates displayed a unique biomarker pattern that was 98% specific to humans. Twenty-nine percent of cattle isolates displayed a unique biomarker that was 97% specific to cattle. Interestingly, even within a related host group (i.e., Family: Canidae [domestic dogs and coyotes]), highly specific SNP biomarkers (98% and 99% specificity for dog and coyotes, respectively) were observed, with 21% of dog E. coli isolates displaying a unique dog biomarker and 61% of coyote isolates displaying a unique coyote biomarker. Application of a supervised learning method, such as logic regression, to DNA sequence analysis at certain intergenic regions demonstrates that some E. coli strains may evolve to become host-specific. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Aportes del Aprendizaje Basado en Problemas (ABP) en la ensenanza de la Fisiologia Animal en un programa de Zootecnia

    NASA Astrophysics Data System (ADS)

    Reinartz-Estrada, Monica

    Based on difficulties observed on the subject of technical-scientific conceptualization and the integration of theory and practice in learning animal physiology for students in the Animal Science program at the National University of Colombia in Medellin, this research paper proposes a problem-based learning strategy founded on the method of Problem Based Learning (PBL), applied specifically to the issues of thermoregulation and physiological stress in domestic animals. In this case study, a sample size of eight students was presented with a pedagogical problem during the first session that would then be solved during the course. In order to evaluate the process, three surveys were conducted called Level Test Formulations (NF) performed at different times of the trial: one before beginning the topic (NF 1), one after three theoretical classes had been given and before beginning the fieldwork (NF 2), and another one after the end of the process (NF 3). Finally, individual interviews were conducted with each student to know the students' perceptions regarding the method. The information obtained was subjected to a qualitative analysis and categorization, using the QDA Miner program which reviewed and coded texts from the surveys and individual interviews, supplemented in turn, by field observation, analyzing the conceptual change, the theory-practice relationship and the correlation between the variables and categories established. Among the main results obtained, it should be noted that following the implementation of PBL in this Animal Physiology course, support for conceptual change was demonstrated and the formulated problem served as a connector between theory and practice. Moreover, there was a fusion of prior knowledge with newly acquired knowledge, meaningful learning, improvement in the level of conceptualization and an increase in the scientificness of definitions; it also led to problem-solving and overcoming epistemological obstacles such as multidisciplinarity and nonlinearity. As a result of this research, it is recommended that this method be evaluated in other topics related to Animal Physiology, in other sciences, in larger sample sizes, as well as to address the issue of evaluation applied directly to this method. Key words: Problem Based Learning (PBL), conceptual change, integration of theory and practice, significatif learning, animal physiology, thermoregulation, physiological stress.

  9. Optimized design and analysis of preclinical intervention studies in vivo

    PubMed Central

    Laajala, Teemu D.; Jumppanen, Mikael; Huhtaniemi, Riikka; Fey, Vidal; Kaur, Amanpreet; Knuuttila, Matias; Aho, Eija; Oksala, Riikka; Westermarck, Jukka; Mäkelä, Sari; Poutanen, Matti; Aittokallio, Tero

    2016-01-01

    Recent reports have called into question the reproducibility, validity and translatability of the preclinical animal studies due to limitations in their experimental design and statistical analysis. To this end, we implemented a matching-based modelling approach for optimal intervention group allocation, randomization and power calculations, which takes full account of the complex animal characteristics at baseline prior to interventions. In prostate cancer xenograft studies, the method effectively normalized the confounding baseline variability, and resulted in animal allocations which were supported by RNA-seq profiling of the individual tumours. The matching information increased the statistical power to detect true treatment effects at smaller sample sizes in two castration-resistant prostate cancer models, thereby leading to saving of both animal lives and research costs. The novel modelling approach and its open-source and web-based software implementations enable the researchers to conduct adequately-powered and fully-blinded preclinical intervention studies, with the aim to accelerate the discovery of new therapeutic interventions. PMID:27480578

  10. Optimized design and analysis of preclinical intervention studies in vivo.

    PubMed

    Laajala, Teemu D; Jumppanen, Mikael; Huhtaniemi, Riikka; Fey, Vidal; Kaur, Amanpreet; Knuuttila, Matias; Aho, Eija; Oksala, Riikka; Westermarck, Jukka; Mäkelä, Sari; Poutanen, Matti; Aittokallio, Tero

    2016-08-02

    Recent reports have called into question the reproducibility, validity and translatability of the preclinical animal studies due to limitations in their experimental design and statistical analysis. To this end, we implemented a matching-based modelling approach for optimal intervention group allocation, randomization and power calculations, which takes full account of the complex animal characteristics at baseline prior to interventions. In prostate cancer xenograft studies, the method effectively normalized the confounding baseline variability, and resulted in animal allocations which were supported by RNA-seq profiling of the individual tumours. The matching information increased the statistical power to detect true treatment effects at smaller sample sizes in two castration-resistant prostate cancer models, thereby leading to saving of both animal lives and research costs. The novel modelling approach and its open-source and web-based software implementations enable the researchers to conduct adequately-powered and fully-blinded preclinical intervention studies, with the aim to accelerate the discovery of new therapeutic interventions.

  11. How can Smartphone-Based Internet Data Support Animal Ecology Fieldtrip?

    NASA Astrophysics Data System (ADS)

    Kurniawan, I. S.; Tapilow, F. S.; Hidayat, T.

    2017-09-01

    Identification and classification skills must be owned by the students. In animal ecology course, the identification and classification skills are necessary to study animals. This experimental study aims to describe the identification and classification skills of students on animal ecology field trip to studying various bird species using smartphone-based internet data. Using Involving 63 students divided into 7 groups for each observation station. Data of birds were sampled using line transect method (5000 meters/station). The results showed the identification and classification skills of students are in sufficient categories. Most students have difficulties because of the limitations of data on the internet about birds. In general, students support the use of smartphones in field trip activities. The results of this study can be used as a reference for the development of learning using smartphones in the future by developing application about birds. The outline, smartphones can be used as a method of alternative learning but needs to be developed for some special purposes.

  12. ASSOCIATIVE CONCEPT LEARNING IN ANIMALS

    PubMed Central

    Zentall, Thomas R.; Wasserman, Edward A.; Urcuioli, Peter J.

    2014-01-01

    Nonhuman animals show evidence for three types of concept learning: perceptual or similarity-based in which objects/stimuli are categorized based on physical similarity; relational in which one object/stimulus is categorized relative to another (e.g., same/different); and associative in which arbitrary stimuli become interchangeable with one another by virtue of a common association with another stimulus, outcome, or response. In this article, we focus on various methods for establishing associative concepts in nonhuman animals and evaluate data documenting the development of associative classes of stimuli. We also examine the nature of the common within-class representation of samples that have been associated with the same reinforced comparison response (i.e., many-to-one matching) by describing manipulations for distinguishing possible representations. Associative concepts provide one foundation for human language such that spoken and written words and the objects they represent become members of a class of interchangeable stimuli. The mechanisms of associative concept learning and the behavioral flexibility it allows, however, are also evident in the adaptive behaviors of animals lacking language. PMID:24170540

  13. Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil.

    PubMed

    Fornazari, Felipe; da Silva, Rodrigo Costa; Richini-Pereira, Virginia Bodelão; Beserra, Hugo Enrique Orsini; Luvizotto, Maria Cecília Rui; Langoni, Helio

    2012-09-01

    Leptospirosis is an infectious disease of worldwide importance. The development of diagnostic techniques allows sick animals to be identified, reservoirs to be eliminated and the disease prevented and controlled. The present study aimed to compare different techniques for diagnosing leptospirosis in sheep. Samples of kidney, liver and blood were collected from 465 animals that originated from a slaughterhouse. The sera were analyzed by the Microscopic Agglutination Test (MAT), and kidney and liver samples of seropositive animals were analyzed using four techniques: bacteriological culture, the Warthin Starry (WS) technique, conventional PCR (cPCR), and quantitative PCR (qPCR). With the MAT, 21 animals were positive (4.5%) to serovars Hardjo (n=12), Hebdomadis (n=5), Sentot (n=2), Wolfii (n=1) and Shermani (n=1). Titers were 100 (n=10), 200 (n=2), 400 (n=6) and 1600 (n=3). No animal was positive by bacteriological culture; four animals were positive by the WS technique in kidney samples; six animals were positive by cPCR in kidney samples; and 11 animals were positive by qPCR, eight of which in kidney samples and three in liver. The bacterial quantification revealed a median of 4.3 bacteria/μL in liver samples and 36.6 bacteria/μL in kidney samples. qPCR presented the highest sensitivity among the techniques, followed by cPCR, the WS technique and bacteriological culture. These results indicate that sheep can carry leptospires of the Sejroe serogroup, and demonstrate the efficiency of quantitative PCR to detect Leptospira spp. in tissue samples. Published by Elsevier B.V.

  14. Detection and Diversity Evaluation of Tetracycline Resistance Genes in Grassland-Based Production Systems in Colombia, South America

    PubMed Central

    Santamaría, Johanna; López, Liliana; Soto, Carlos Yesid

    2011-01-01

    Grassland-based production systems use ∼26% of land surface on earth. However, there are no evaluations of these systems as a source of antibiotic pollution. This study was conducted to evaluate the presence, diversity, and distribution of tetracycline resistance genes in the grasslands of the Colombian Andes, where administration of antibiotics to animals is limited to treat disease and growth promoters are not included in animals’ diet. Animal (ruminal fluid and feces) and environmental (soil and water) samples were collected from different dairy cattle farms and evaluated by PCR for the genes tet(M), tet(O), tetB(P), tet(Q), tet(W), tet(S), tet(T), otr(A), which encode ribosomal protection proteins (RPPs), and the genes tet(A), tet(B), tet(D), tet(H), tet(J), and tet(Z), encoding efflux pumps. A wide distribution and high frequency for genes tet(W) and tet(Q) were found in both sample types. Genes tet(O) and tetB(P), detected in high frequencies in feces, were detected in low frequencies or not detected at all in the environment. Other genes encoding RPPs, such as tet(M), tet(S), and tet(T), were detected at very low frequencies and restricted distributions. Genes encoding efflux pumps were not common in this region, and only two of them, tet(B) and tet(Z), were detected. DGGE–PCR followed by comparative sequence analysis of tet(W) and tet(Q) showed that the sequences detected in animals did not differ from those coming from soil and water. Finally, the farms sampled in this study showed more than 50% similarity in relation to the tet genes detected. In conclusion, there was a remarkable presence of tet genes in these production systems and, although not all genes detected in animal reservoirs were detected in the environment, there is a predominant distribution of tet(W) and tet(Q) in both animal and environmental reservoirs. Sequence similarity analysis suggests the transmission of these genes from animals to the environment. PMID:22174707

  15. Quantitative determination of carcinogenic mycotoxins in human and animal biological matrices and animal-derived foods using multi-mycotoxin and analyte-specific high performance liquid chromatography-tandem mass spectrometric methods.

    PubMed

    Cao, Xiaoqin; Li, Xiaofei; Li, Jian; Niu, Yunhui; Shi, Lu; Fang, Zhenfeng; Zhang, Tao; Ding, Hong

    2018-01-15

    A sensitive and reliable multi-mycotoxin-based method was developed to identify and quantify several carcinogenic mycotoxins in human blood and urine, as well as edible animal tissues, including muscle and liver tissue from swine and chickens, using liquid chromatography-tandem mass spectrometry (LC-MS/MS). For the toxicokinetic studies with individual mycotoxins, highly sensitive analyte-specific LC-MS/MS methods were developed for rat plasma and urine. Sample purification consisted of a rapid 'dilute and shoot' approach in urine samples, a simple 'dilute, evaporate and shoot' approach in plasma samples and a 'QuEChERS' procedure in edible animal tissues. The multi-mycotoxin and analyte-specific methods were validated in-house: The limits of detection (LOD) for the multi-mycotoxin and analyte-specific methods ranged from 0.02 to 0.41 μg/kg (μg/L) and 0.01 to 0.19 μg/L, respectively, and limits of quantification (LOQ) between 0.10 to 1.02 μg/kg (μg/L) and 0.09 to 0.47 μg/L, respectively. Apparent recoveries of the samples spiked with 0.25 to 4 μg/kg (μg/L) ranged from 60.1% to 109.8% with relative standard deviations below 15%. The methods were successfully applied to real samples. To the best of our knowledge, this is the first study carried out using a small group of patients from the Chinese population with hepatocellular carcinoma to assess their exposure to carcinogenic mycotoxins using biomarkers. Finally, the multi-mycotoxin method is a useful analytical method for assessing exposure to mycotoxins edible in animal tissues. The analyte-specific methods could be useful during toxicokinetic and toxicological studies. Copyright © 2017. Published by Elsevier B.V.

  16. DNA barcode based wildlife forensics for resolving the origin of claw samples using a novel primer cocktail.

    PubMed

    Khedkar, Gulab D; Abhayankar, Shil Bapurao; Nalage, Dinesh; Ahmed, Shaikh Nadeem; Khedkar, Chandraprakash D

    2016-11-01

    Excessive wildlife hunting for commercial purposes can have negative impacts on biodiversity and may result in species extinction. To ensure compliance with legal statutes, forensic identification approaches relying on molecular markers may be used to identify the species of origin of animal material from hairs, claw, blood, bone, or meat. Using this approach, DNA sequences from the COI "barcoding" gene have been used to identify material from a number of domesticated animal species. However, many wild species of carnivores still present great challenges in generating COI barcodes using standard "universal" primer pairs. In the work presented here, the mitochondrial COI gene was successfully amplified using a novel primer cocktail, and the products were sequenced to determine the species of twenty one unknown samples of claw material collected as part of forensic wildlife case investigations. Sixteen of the unknown samples were recognized to have originated from either Panthera leo or P. pardus individuals. The remaining five samples could be identified only to the family level due to the absence of reference animal sequences. This is the first report on the use of COI sequences for the identification of P. pardus and P. leo from claw samples as part of forensic investigations in India. The study also highlights the need for adequate reference material to aid in the resolution of suspected cases of illegal wildlife harvesting.

  17. Levels in food and beverages and daily intake of perfluorinated compounds in Norway.

    PubMed

    Haug, Line Småstuen; Salihovic, Samira; Jogsten, Ingrid Ericson; Thomsen, Cathrine; van Bavel, Bert; Lindström, Gunilla; Becher, Georg

    2010-08-01

    Perfluorinated compounds (PFCs) have been determined in 21 samples of selected food and beverages such as meat, fish, bread, vegetables, milk, drinking water and tea from the Norwegian marked. Up to 12 different PFCs were detected in the samples. Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) were found in concentrations similar to or lower than what has been observed in other studies world-wide. Differences in the relative proportion of PFOA and PFOS between samples of animal origin and samples of non-animal origin were observed and support findings that PFOS has a higher bioaccumulation potential in animals than PFOA. Based on these 21 measurements and consumption data for the general Norwegian population, a rough estimate of the total dietary intake of PFCs was found to be around 100 ng d(-1). PFOA and PFOS contributed to about 50% of the total intake. When dividing the population in gender and age groups, estimated intakes were decreasing with increasing age and were higher in males than females. The estimated intakes of PFOS and PFOA in the present study are lower than what has been reported in studies from Spain, Germany, United Kingdom, Canada and Japan. This study illustrates that by improving the analytical methods for determination of PFC in food samples, a broad range of compounds can be detected, which is important when assessing dietary exposure. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  18. PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp.

    PubMed Central

    LOPES, Estela Gallucci; GERALDO, Carlos Alberto; MARCILI, Arlei; SILVA, Ricardo Duarte; KEID, Lara Borges; OLIVEIRA, Trícia Maria Ferreira da Silva; SOARES, Rodrigo Martins

    2016-01-01

    In visceral leishmaniasis, the detection of the agent is of paramount importance to identify reservoirs of infection. Here, we evaluated the diagnostic attributes of PCRs based on primers directed to cytochrome-B (cytB), cytochrome-oxidase-subunit II (coxII), cytochrome-C (cytC), and the minicircle-kDNA. Although PCRs directed to cytB, coxII, cytC were able to detect different species of Leishmania, and the nucleotide sequence of their amplicons allowed the unequivocal differentiation of species, the analytical and diagnostic sensitivity of these PCRs were much lower than the analytical and diagnostic sensitivity of the kDNA-PCR. Among the 73 seropositive animals, the asymptomatic dogs had spleen and bone marrow samples collected and tested; only two animals were positive by PCRs based on cytB, coxII, and cytC, whereas 18 were positive by the kDNA-PCR. Considering the kDNA-PCR results, six dogs had positive spleen and bone marrow samples, eight dogs had positive bone marrow results but negative results in spleen samples and, in four dogs, the reverse situation occurred. We concluded that PCRs based on cytB, coxII, and cytC can be useful tools to identify Leishmania species when used in combination with automated sequencing. The discordance between the results of the kDNA-PCR in bone marrow and spleen samples may indicate that conventional PCR lacks sensitivity for the detection of infected dogs. Thus, primers based on the kDNA should be preferred for the screening of infected dogs. PMID:27253743

  19. 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.

  20. 9 CFR 592.450 - Procedures for selecting appeal samples.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Procedures for selecting appeal samples. 592.450 Section 592.450 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION VOLUNTARY INSPECTION OF EGG PRODUCTS Appeals § 592.450...

  1. Analysing designed experiments in distance sampling

    Treesearch

    Stephen T. Buckland; Robin E. Russell; Brett G. Dickson; Victoria A. Saab; Donal N. Gorman; William M. Block

    2009-01-01

    Distance sampling is a survey technique for estimating the abundance or density of wild animal populations. Detection probabilities of animals inherently differ by species, age class, habitats, or sex. By incorporating the change in an observer's ability to detect a particular class of animals as a function of distance, distance sampling leads to density estimates...

  2. Use of molecular and milk production information for the cost-effective diagnosis of bovine viral diarrhoea infection in New Zealand dairy cattle.

    PubMed

    Hill, F I; Reichel, M P; Tisdall, D J

    2010-04-21

    An increase in veterinary and farmer interest in bovine viral diarrhoea (BVD) in New Zealand over recent years led to requests for cost-effective identification of BVD virus (BVDV) infected herds and individuals. This study was undertaken to determine if the use of real-time reverse transcriptase polymerase chain reaction (RT-PCR) technology and dairy cow production data could identify persistently infected (PI) animals in milking herds. Milk samples were collected from the vats of dairy herds and tested for the presence of BVDV by RT-PCR till four herds were found containing PI animals. Individual serum samples were then collected from every cow in the herd and tested by both RT-PCR and antigen capture enzyme-linked immunosorbent assays (ACE) to identify the PI animals. Individual animal testing found 1/223, 1/130, 2/800 and 1/275 PI's respectively in the four herds. Based on these results a maximum pool size of 400 cows contributing to the bulk tank milk was selected. After removal of the PI from the herds, further bulk milk samples were shown to be BVDV negative by RT-PCR. All the PI animals identified by this method were found in the lowest producing 10-20% of herd. This approach of targeted testing of dairy herds using PCR technology, in conjunction with animal production information, markedly reduced the cost of diagnostic testing for BVDV in dairy herds in New Zealand. Questionnaire follow-up on 81 BVDV-positive herds (15% of those tested) indicated the stratification approach identified milking PIs successfully over 90% of the time and reduced the number of individual tests to 12% of the milking herd. Copyright 2009 Elsevier B.V. All rights reserved.

  3. Prevalence of Campylobacter jejuni and Campylobacter coli in captive wildlife species of India

    PubMed Central

    Prince Milton, A. A.; Agarwal, R. K.; Priya, G. B.; Saminathan, M.; Aravind, M.; Reddy, A.; Athira, C. K.; Anjay; Ramees, T. P.; Dhama, K.; Sharma, A. K.; Kumar, A.

    2017-01-01

    Campylobacteriosis is an important zoonotic disease and the prevalence of Campylobacter is largely unknown in the wildlife of India. A total of 370 samples, comprising of 314 fresh faecal samples from apparently healthy captive wild animals and birds, 30 stool swabs from animal care takers and 26 samples of the animals’ food and water were collected from G. B. Pant High Altitude Zoo, Nainital, Kanpur Zoo, Wildlife Park, IVRI and the Post Graduate Research Institute in Animal Sciences (PGRIAS), Chennai, Tamilnadu from August 2014 to May 2015. Samples were processed for cultural isolation, direct PCR and multiplex PCR for species confirmation. To decipher the genetic diversity, the 16S rRNA gene was amplified, sequenced and analyzed. Based on isolation, the overall occurrence rate of Campylobacter spp. was 0.8% (3/370), being 2.94% (3/102) for captive wild birds. Three Campylobacter jejuni were isolated from silver pheasants, lady amherest pheasants and saras cranes. Direct PCR assay showed the overall occurrence rate of Campylobacter spp. to be 4.77% (15/315), being 1.58% (2/126) for captive wild ruminants, 5.81% (5/86) for non-ruminants and 7.84% (8/102) for birds. All the isolates were identified as C. jejuni. PMID:29163646

  4. Sampling designs matching species biology produce accurate and affordable abundance indices

    PubMed Central

    Farley, Sean; Russell, Gareth J.; Butler, Matthew J.; Selinger, Jeff

    2013-01-01

    Wildlife biologists often use grid-based designs to sample animals and generate abundance estimates. Although sampling in grids is theoretically sound, in application, the method can be logistically difficult and expensive when sampling elusive species inhabiting extensive areas. These factors make it challenging to sample animals and meet the statistical assumption of all individuals having an equal probability of capture. Violating this assumption biases results. Does an alternative exist? Perhaps by sampling only where resources attract animals (i.e., targeted sampling), it would provide accurate abundance estimates more efficiently and affordably. However, biases from this approach would also arise if individuals have an unequal probability of capture, especially if some failed to visit the sampling area. Since most biological programs are resource limited, and acquiring abundance data drives many conservation and management applications, it becomes imperative to identify economical and informative sampling designs. Therefore, we evaluated abundance estimates generated from grid and targeted sampling designs using simulations based on geographic positioning system (GPS) data from 42 Alaskan brown bears (Ursus arctos). Migratory salmon drew brown bears from the wider landscape, concentrating them at anadromous streams. This provided a scenario for testing the targeted approach. Grid and targeted sampling varied by trap amount, location (traps placed randomly, systematically or by expert opinion), and traps stationary or moved between capture sessions. We began by identifying when to sample, and if bears had equal probability of capture. We compared abundance estimates against seven criteria: bias, precision, accuracy, effort, plus encounter rates, and probabilities of capture and recapture. One grid (49 km2 cells) and one targeted configuration provided the most accurate results. Both placed traps by expert opinion and moved traps between capture sessions, which raised capture probabilities. The grid design was least biased (−10.5%), but imprecise (CV 21.2%), and used most effort (16,100 trap-nights). The targeted configuration was more biased (−17.3%), but most precise (CV 12.3%), with least effort (7,000 trap-nights). Targeted sampling generated encounter rates four times higher, and capture and recapture probabilities 11% and 60% higher than grid sampling, in a sampling frame 88% smaller. Bears had unequal probability of capture with both sampling designs, partly because some bears never had traps available to sample them. Hence, grid and targeted sampling generated abundance indices, not estimates. Overall, targeted sampling provided the most accurate and affordable design to index abundance. Targeted sampling may offer an alternative method to index the abundance of other species inhabiting expansive and inaccessible landscapes elsewhere, provided their attraction to resource concentrations. PMID:24392290

  5. Occurrence of enrofloxacin in overflows from animal lot and residential sewage lagoons and a receiving-stream.

    PubMed

    Ikem, Abua; Lin, Chung-Ho; Broz, Bob; Kerley, Monty; Thi, Ho Le

    2017-10-01

    Enrofloxacin (ENRO), a fluoroquinolone, was quantified in overflows from an animal lot and residential sewage lagoons and in a receiving-stream (Gans Creek). The concentrations of ENRO in samples was determined by high-performance liquid chromatography - tandem mass spectrometry. In total, ninety samples including duplicates were analyzed during several monthly sampling campaigns. The samples collected represented the residential sewage lagoon overflow (RLO), animal lot lagoon overflow (ALLO), the combined overflows (RLO and ALLO), and Gans Creek (upstream, midstream and downstream positions). The frequency of detection of ENRO was 90% for RLO and 100% for both ALLO and Gans Creek. The highest concentration of ENRO (0.44 μg/L) was found in ALLO sample collected during high precipitation. ENRO levels found in RLO samples ranged from < LOQ to 259 ng/L and the highest value observed also coincided with high flow. The levels of ENRO found in Gans Creek ranged from 17-216 ng/L. A preliminary ecotoxicological assessment was conducted through calculation of the risk quotients (RQs) for organisms based on the ratio of the measured environmental concentrations in this study to the predicted-no-effect-concentrations (acute and chronic effect) data. From the RQs, high risks were observed for Microcystis aeruginosa (cyanobacteria; RQ = 4.4); Anabaena flosaquae (cyanobacteria; RQ = 1.3); and Lemna minor (aquatic vascular plant; RQ = 2.0). The long-term effects of mixtures of PHCs on Gans Creek watershed are probable.

  6. Veterinary extension on sampling techniques related to heartwater research.

    PubMed

    Steyn, H C; McCrindle, C M E; Du Toit, D

    2010-09-01

    Heartwater, a tick-borne disease caused by Ehrlichia ruminantium, is considered to be a significant cause of mortality amongst domestic and wild ruminants in South Africa. The main vector is Amblyomma hebraeum and although previous epidemiological studies have outlined endemic areas based on mortalities, these have been limited by diagnostic methods which relied mainly on positive brain smears. The indirect fluorescent antibody test (IFA) has a low specificity for heartwater organisms as it cross-reacts with some other species. Since the advent of biotechnology and genomics, molecular epidemiology has evolved using the methodology of traditional epidemiology coupled with the new molecular techniques. A new quantitative real-time polymerase chain reaction (qPCR) test has been developed for rapid and accurate diagnosis of heartwater in the live animal. This method can also be used to survey populations of A. hebraeum ticks for heartwater. Sampling whole blood and ticks for this qPCR differs from routine serum sampling, which is used for many serological tests. Veterinary field staff, particularly animal health technicians, are involved in surveillance and monitoring of controlled and other diseases of animals in South Africa. However, it was found that the sampling of whole blood was not done correctly, probably because it is a new sampling technique specific for new technology, where the heartwater organism is much more labile than the serum antibodies required for other tests. This qPCR technique is highly sensitive and can diagnose heartwater in the living animal within 2 hours, in time to treat it. Poor sampling techniques that decrease the sensitivity of the test will, however, result in a false negative diagnosis. This paper describes the development of a skills training programme for para-veterinary field staff, to facilitate research into the molecular epidemiology of heartwater in ruminants and eliminate any sampling bias due to collection errors. Humane handling techniques were also included in the training, in line with the current focus on improved livestock welfare.

  7. Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community.

    PubMed

    Tapio, Ilma; Shingfield, Kevin J; McKain, Nest; Bonin, Aurélie; Fischer, Daniel; Bayat, Ali R; Vilkki, Johanna; Taberlet, Pierre; Snelling, Timothy J; Wallace, R John

    2016-01-01

    Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint.

  8. Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community

    PubMed Central

    Tapio, Ilma; Shingfield, Kevin J.; McKain, Nest; Bonin, Aurélie; Fischer, Daniel; Bayat, Ali R.; Vilkki, Johanna; Taberlet, Pierre; Snelling, Timothy J.; Wallace, R. John

    2016-01-01

    Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint. PMID:26986467

  9. The bovine lung in biomedical research: visually guided bronchoscopy, intrabronchial inoculation and in vivo sampling techniques.

    PubMed

    Prohl, Annette; Ostermann, Carola; Lohr, Markus; Reinhold, Petra

    2014-07-03

    There is an ongoing search for alternative animal models in research of respiratory medicine. Depending on the goal of the research, large animals as models of pulmonary disease often resemble the situation of the human lung much better than mice do. Working with large animals also offers the opportunity to sample the same animal repeatedly over a certain course of time, which allows long-term studies without sacrificing the animals. The aim was to establish in vivo sampling methods for the use in a bovine model of a respiratory Chlamydia psittaci infection. Sampling should be performed at various time points in each animal during the study, and the samples should be suitable to study the host response, as well as the pathogen under experimental conditions. Bronchoscopy is a valuable diagnostic tool in human and veterinary medicine. It is a safe and minimally invasive procedure. This article describes the intrabronchial inoculation of calves as well as sampling methods for the lower respiratory tract. Videoendoscopic, intrabronchial inoculation leads to very consistent clinical and pathological findings in all inoculated animals and is, therefore, well-suited for use in models of infectious lung disease. The sampling methods described are bronchoalveolar lavage, bronchial brushing and transbronchial lung biopsy. All of these are valuable diagnostic tools in human medicine and could be adapted for experimental purposes to calves aged 6-8 weeks. The samples obtained were suitable for both pathogen detection and characterization of the severity of lung inflammation in the host.

  10. Development of an ELA-DRA gene typing method based on pyrosequencing technology.

    PubMed

    Díaz, S; Echeverría, M G; It, V; Posik, D M; Rogberg-Muñoz, A; Pena, N L; Peral-García, P; Vega-Pla, J L; Giovambattista, G

    2008-11-01

    The polymorphism of equine lymphocyte antigen (ELA) class II DRA gene had been detected by polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) and reference strand-mediated conformation analysis. These methodologies allowed to identify 11 ELA-DRA exon 2 sequences, three of which are widely distributed among domestic horse breeds. Herein, we describe the development of a pyrosequencing-based method applicable to ELA-DRA typing, by screening samples from eight different horse breeds previously typed by PCR-SSCP. This sequence-based method would be useful in high-throughput genotyping of major histocompatibility complex genes in horses and other animal species, making this system interesting as a rapid screening method for animal genotyping of immune-related genes.

  11. Rapid and Specific Detection of Salmonella spp. in Animal Feed Samples by PCR after Culture Enrichment

    PubMed Central

    Löfström, Charlotta; Knutsson, Rickard; Axelsson, Charlotta Engdahl; Rådström, Peter

    2004-01-01

    A PCR procedure has been developed for routine analysis of viable Salmonella spp. in feed samples. The objective was to develop a simple PCR-compatible enrichment procedure to enable DNA amplification without any sample pretreatment such as DNA extraction or cell lysis. PCR inhibition by 14 different feed samples and natural background flora was circumvented by the use of the DNA polymerase Tth. This DNA polymerase was found to exhibit a high level of resistance to PCR inhibitors present in these feed samples compared to DyNAzyme II, FastStart Taq, Platinum Taq, Pwo, rTth, Taq, and Tfl. The specificity of the Tth assay was confirmed by testing 101 Salmonella and 43 non-Salmonella strains isolated from feed and food samples. A sample preparation method based on culture enrichment in buffered peptone water and DNA amplification with Tth DNA polymerase was developed. The probability of detecting small numbers of salmonellae in feed, in the presence of natural background flora, was accurately determined and found to follow a logistic regression model. From this model, the probability of detecting 1 CFU per 25 g of feed in artificially contaminated soy samples was calculated and found to be 0.81. The PCR protocol was evaluated on 155 naturally contaminated feed samples and compared to an established culture-based method, NMKL-71. Eight percent of the samples were positive by PCR, compared with 3% with the conventional method. The reasons for the differences in sensitivity are discussed. Use of this method in the routine analysis of animal feed samples would improve safety in the food chain. PMID:14711627

  12. The Assessment of Animal Welfare in British Zoos by Government-Appointed Inspectors

    PubMed Central

    Draper, Chris; Harris, Stephen

    2012-01-01

    Simple Summary Since 1984, British zoos have been required to meet the animal welfare standards set out under the Zoo Licensing Act 1981. Zoos are regularly assessed by government-appointed inspectors, who report on animal welfare standards in each zoo. This is the first analysis of those reports from a representative sample of British zoos. We highlight a number of concerns about the inspection process itself, and identify areas where changes would lead to improvements in both the inspection process and our ability to monitor animal welfare standards in zoos. Abstract We analysed the reports of government-appointed inspectors from 192 zoos between 2005–2008 to provide the first review of how animal welfare was assessed in British zoos since the enactment of the Zoo Licensing Act 1981. We examined the effects of whether or not a veterinarian was included in the inspection team, type of inspection, licence status of the zoo and membership of a zoo association on the inspectors’ assessments of animal welfare standards in five areas that approximate to the Five Freedoms. At least 11% of full licence inspections did not comply with the legal requirement for two inspectors. The inspectors’ reports were unclear as to how animal welfare was assessed, whether all animals or only a sub-sample had been inspected, and were based predominantly on welfare inputs rather than outcomes. Of 9,024 animal welfare assessments across the 192 zoos, 7,511 (83%) were graded as meeting the standards, 782 (9%) as substandard and the rest were not graded. Of the 192 zoos, 47 (24%) were assessed as meeting all the animal welfare standards. Membership of a zoo association was not associated with a higher overall assessment of animal welfare standards, and specialist collections such as Farm Parks and Other Bird collections performed least well. We recommend a number of changes to the inspection process that should lead to greater clarity in the assessment of animal welfare in British zoos. PMID:26487161

  13. 9 CFR 355.26 - Samples of certified products, ingredients, etc., to be taken for examination.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Samples of certified products, ingredients, etc., to be taken for examination. 355.26 Section 355.26 Animals and Animal Products FOOD SAFETY..., CATS, AND OTHER CARNIVORA; INSPECTION, CERTIFICATION, AND IDENTIFICATION AS TO CLASS, QUALITY, QUANTITY...

  14. 9 CFR 355.26 - Samples of certified products, ingredients, etc., to be taken for examination.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Samples of certified products, ingredients, etc., to be taken for examination. 355.26 Section 355.26 Animals and Animal Products FOOD SAFETY..., CATS, AND OTHER CARNIVORA; INSPECTION, CERTIFICATION, AND IDENTIFICATION AS TO CLASS, QUALITY, QUANTITY...

  15. Determination of Milk Fat Adulteration with Vegetable Oils and Animal Fats by Gas Chromatographic Analysis.

    PubMed

    Kim, Jin-Man; Kim, Ha-Jung; Park, Jung-Min

    2015-09-01

    This study assessed the potential application of gas chromatography (GC) in detecting milk fat (MF) adulteration with vegetable oils and animal fats and of characterizing samples by fat source. One hundred percent pure MF was adulterated with different vegetable oils and animal fats at various concentrations (0%, 10%, 30%, 50%, 70%, and 90%). GC was used to obtain the fatty acid (FA) profiles, triacylglycerol (TG) contents, and cholesterol contents. The pure MF and the adulterated MF samples were discriminated based on the total concentrations of saturated FAs and on the 2 major FAs (oleic acid [C18:1n9c] and linoleic acid [C18:2n6c], TGs [C52 and C54], and cholesterol contents using statistical analysis to compared difference. These bio-markers enabled the detection of as low as 10% adulteration of non-MF into 100% pure MF. The study demonstrated the high potential of GC to rapidly detect MF adulteration with vegetable and animal fats, and discriminate among commercial butter and milk products according to the fat source. These data can be potentially useful in detecting foreign fats in these butter products. Furthermore, it is important to consider that several individual samples should be analyzed before coming to a conclusion about MF authenticity. © 2015 Institute of Food Technologists®

  16. Epidemiology of foot-and-mouth disease in Landhi Dairy Colony, Pakistan, the world largest Buffalo colony

    PubMed Central

    Klein, Joern; Hussain, Manzoor; Ahmad, Munir; Afzal, Muhammad; Alexandersen, Soren

    2008-01-01

    Background Foot-and-mouth disease (FMD) is endemic in Pakistan and causes huge economic losses. This work focus on the Landhi Dairy Colony (LDC), located in the suburbs of Karachi. LDC is the largest Buffalo colony in the world, with more than 300,000 animals (around 95% buffaloes and 5% cattle, as well as an unknown number of sheep and goats). Each month from April 2006 to April 2007 we collected mouth-swabs from apparently healthy buffaloes and cattle, applying a convenient sampling based on a two-stage random sampling scheme, in conjunction with participatory information from each selected farm. Furthermore, we also collected epithelium samples from animals with clinical disease, as well as mouth-swabs samples from those farms. In addition, we analysed a total of 180 serum samples randomly collecting 30 samples each month at the local slaughterhouse, from October 2006 to March 2007. Samples have been screened for FMDV by real-time RT-PCR and the partial or full 1D coding region of selected isolates has been sequenced. Serum samples have been analysed by applying serotype-specific antibody ELISA and non-structural proteins (NSP) antibody ELISA. Results FMDV infection prevalence at aggregate level shows an endemic occurrence of FMDV in the colony, with peaks in August 2006, December 2006 and February 2007 to March 2007. A significant association of prevalence peaks to the rainy seasons, which includes the coldest time of the year and the muslimic Eid-festival, has been demonstrated. Participatory information indicated that 88% of all questioned farmers vaccinate their animals. Analysis of the serum samples showed high levels of antibodies for serotypes O, A, Asia 1 and C. The median endpoint-titre for all tested serotypes, except serotype C, in VNT titration is at a serum dilution of equal or above 1/100. All 180 serum samples collected have been tested for antibodies against the non-structural proteins and all but four have been found positive. Out of the 106 swab-samples from apparently healthy and affected animals positive in real-time RT-PCR, we sequenced the partial or full 1D coding region from 58 samples. In addition we sequenced the full 1D coding region of 17 epithelium samples from animals with clinical signs of FMD. From all sequenced samples, swabs and epithelium, 19 belong to the regional PanAsia II lineage of serotype O and 56 to the A/Iran/2005 lineage of serotype A. Conclusion For an effective and realisable FMD control program in LDC, we suggest to introduce a twice annually mass vaccination of all buffaloes and cattle in the colony. These mass vaccinations should optimally take place shortly before the beginning of the two rainy periods, e.g. in June and September. Those vaccinations should, in our opinion, be in addition to the already individually performed vaccinations of single animals, as the latter usually targets only newly introduced animals. This suggested combination of mass vaccination of all large ruminants with the already performed individually vaccination should provide a continuous high level of herd immunity in the entire colony. Vaccines used for this purpose should contain the matching vaccine strains, i.e. as our results indicate antigens for A/Iran/2005 and the regional type of serotype O (PanAsia II), but also antigens of the, in this world region endemic, Asia 1 lineage should be included. In the long term it will be important to control the vaccine use, so that subclinical FMD will be avoided. PMID:18445264

  17. Molecular detection of fungi of public health importance in wild animals from Southern Brazil.

    PubMed

    Losnak, Debora O; Rocha, Francielle R; Almeida, Barbara S; Batista, Keila Z S; Althoff, Sérgio L; Haupt, Josiane; Ruiz, Luciana S; Anversa, Laís; Lucheis, Simone B; Paiz, Laís M; Donalisio, Maria Rita; Richini Pereira, Virginia B

    2018-07-01

    Some animals have an important relationship with fungal infections, and searching for pathogens in animal samples may be an opportunity for eco-epidemiological research. Since studies involving wildlife are generally restricted, using samples from road kills is an alternative. The aim of this study was to verify whether pathogenic fungi of public health importance occur in wildlife road kills from Santa Catarina State, Brazil. Organ samples (n = 1063) from 297 animals were analysed according to Polymerase Chain Reaction (PCR) using universal primers to detect fungi in general and, subsequently, using primers specific to Paracoccidioides brasiliensis, Histoplasma capsulatum and Cryptococcus spp. There were 102 samples positive for fungal species. Eight samples were positive for P. brasiliensis, three samples were positive for Cryptococcus spp. and one sample had coinfection by these two fungi. No sample was positive for Histoplasma spp. according to the molecular detection. Genetic sequencing allowed the identification of Fungal sp. in 89 samples, Cryptococcus neoformans in two samples and Aspergillus penicillioides in three samples. This study shows the importance of wild animals in the epidemiology of fungal infections and assists in the mapping of pathogen occurrence in a region that was not previously evaluated. © 2018 Blackwell Verlag GmbH.

  18. Dual Combined Real-Time Reverse Transcription Polymerase Chain Reaction Assay for the Diagnosis of Lyssavirus Infection.

    PubMed

    Dacheux, Laurent; Larrous, Florence; Lavenir, Rachel; Lepelletier, Anthony; Faouzi, Abdellah; Troupin, Cécile; Nourlil, Jalal; Buchy, Philippe; Bourhy, Herve

    2016-07-01

    The definitive diagnosis of lyssavirus infection (including rabies) in animals and humans is based on laboratory confirmation. The reference techniques for post-mortem rabies diagnosis are still based on direct immunofluorescence and virus isolation, but molecular techniques, such as polymerase chain reaction (PCR) based methods, are increasingly being used and now constitute the principal tools for diagnosing rabies in humans and for epidemiological analyses. However, it remains a key challenge to obtain relevant specificity and sensitivity with these techniques while ensuring that the genetic diversity of lyssaviruses does not compromise detection. We developed a dual combined real-time reverse transcription polymerase chain reaction (combo RT-qPCR) method for pan-lyssavirus detection. This method is based on two complementary technologies: a probe-based (TaqMan) RT-qPCR for detecting the RABV species (pan-RABV RT-qPCR) and a second reaction using an intercalating dye (SYBR Green) to detect other lyssavirus species (pan-lyssa RT-qPCR). The performance parameters of this combined assay were evaluated with a large panel of primary animal samples covering almost all the genetic variability encountered at the viral species level, and they extended to almost all lyssavirus species characterized to date. This method was also evaluated for the diagnosis of human rabies on 211 biological samples (positive n = 76 and negative n = 135) including saliva, skin and brain biopsies. It detected all 41 human cases of rabies tested and confirmed the sensitivity and the interest of skin biopsy (91.5%) and saliva (54%) samples for intra-vitam diagnosis of human rabies. Finally, this method was successfully implemented in two rabies reference laboratories in enzootic countries (Cambodia and Morocco). This combined RT-qPCR method constitutes a relevant, useful, validated tool for the diagnosis of rabies in both humans and animals, and represents a promising tool for lyssavirus surveillance.

  19. Dual Combined Real-Time Reverse Transcription Polymerase Chain Reaction Assay for the Diagnosis of Lyssavirus Infection

    PubMed Central

    Lavenir, Rachel; Lepelletier, Anthony; Faouzi, Abdellah; Troupin, Cécile; Nourlil, Jalal; Buchy, Philippe; Bourhy, Herve

    2016-01-01

    The definitive diagnosis of lyssavirus infection (including rabies) in animals and humans is based on laboratory confirmation. The reference techniques for post-mortem rabies diagnosis are still based on direct immunofluorescence and virus isolation, but molecular techniques, such as polymerase chain reaction (PCR) based methods, are increasingly being used and now constitute the principal tools for diagnosing rabies in humans and for epidemiological analyses. However, it remains a key challenge to obtain relevant specificity and sensitivity with these techniques while ensuring that the genetic diversity of lyssaviruses does not compromise detection. We developed a dual combined real-time reverse transcription polymerase chain reaction (combo RT-qPCR) method for pan-lyssavirus detection. This method is based on two complementary technologies: a probe-based (TaqMan) RT-qPCR for detecting the RABV species (pan-RABV RT-qPCR) and a second reaction using an intercalating dye (SYBR Green) to detect other lyssavirus species (pan-lyssa RT-qPCR). The performance parameters of this combined assay were evaluated with a large panel of primary animal samples covering almost all the genetic variability encountered at the viral species level, and they extended to almost all lyssavirus species characterized to date. This method was also evaluated for the diagnosis of human rabies on 211 biological samples (positive n = 76 and negative n = 135) including saliva, skin and brain biopsies. It detected all 41 human cases of rabies tested and confirmed the sensitivity and the interest of skin biopsy (91.5%) and saliva (54%) samples for intra-vitam diagnosis of human rabies. Finally, this method was successfully implemented in two rabies reference laboratories in enzootic countries (Cambodia and Morocco). This combined RT-qPCR method constitutes a relevant, useful, validated tool for the diagnosis of rabies in both humans and animals, and represents a promising tool for lyssavirus surveillance. PMID:27380028

  20. Osseointegration of alumina bioceramic granules: A comparative experimental study

    NASA Astrophysics Data System (ADS)

    Rerikh, V. V.; Avetisyan, A. R.; Zaydman, A. M.; Anikin, K. A.; Bataev, V. A.; Nikulina, A. A.; Sadovoy, M. A.; Aronov, A. M.; Semantsova, E. S.

    2016-08-01

    To perform a comparative analysis of osseointegration of bioceramic alumina-based granules, hydroxyapatite-based granules, and deproteinized bone granules. The experiment was conducted on 52 adult male Kyoto-Wistar rats weighing 350 to 520 g. The animals were divided into five matched groups that differed only in the type of an implanted material. The granules were implanted in the lumbar vertebral bodies and in the distal right femur of each laboratory animal. Two months after surgery, the animals were euthanized, followed by tissue sampling for morphological studies. An examination of specimens from the groups with implanted alumina granules revealed the newly formed trabecular bone with remodeling signs. The bone tissue filled the intragranular space, tightly adhering to the granule surface. There was no connective tissue capsule on the border between bone tissue and alumina granules. Cylindrical bioceramic alumina-based granules with an open internal channel have a higher strength surpassing than that of analogs and the osseointegration ability close to that of hydroxyapatite and deproteinized bone granules.

  1. Validation of a near infrared microscopy method for the detection of animal products in feedingstuffs: results of a collaborative study.

    PubMed

    Boix, A; Fernández Pierna, J A; von Holst, C; Baeten, V

    2012-01-01

    The performance characteristics of a near infrared microscopy (NIRM) method, when applied to the detection of animal products in feedingstuffs, were determined via a collaborative study. The method delivers qualitative results in terms of the presence or absence of animal particles in feed and differentiates animal from vegetable feed ingredients on the basis of the evaluation of near infrared spectra obtained from individual particles present in the sample. The specificity ranged from 86% to 100%. The limit of detection obtained on the analysis of the sediment fraction, prepared as for the European official method, was 0.1% processed animal proteins (PAPs) in feed, since all laboratories correctly identified the positive samples. This limit has to be increased up to 2% for the analysis of samples which are not sedimented. The required sensitivity for the official control is therefore achieved in the analysis of the sediment fraction of the samples where the method can be applied for the detection of the presence of animal meal. Criteria for the classification of samples, when fewer than five spectra are found, as being of animal origin needs to be set up in order to harmonise the approach taken by the laboratories when applying NIRM for the detection of the presence of animal meal in feed.

  2. Comparative value of blood and skin samples for diagnosis of spotted fever group rickettsial infection in model animals.

    PubMed

    Levin, Michael L; Snellgrove, Alyssa N; Zemtsova, Galina E

    2016-07-01

    The definitive diagnosis of spotted fever group (SFG) rickettsioses in humans is challenging due to the retrospective nature and cross reactivity of the serological methods and the absence of reliable and consistent samples for molecular diagnostics. Existing data indicate the transient character of bacteremia in experimentally infected animals. The ability of arthropod vectors to acquire rickettsial infection from the laboratory animals in the absence of systemic infection and known tropism of rickettsial agents to endothelial cells of peripheral blood vessels underline the importance of local infection and consequently the diagnostic potential of skin samples. In order to evaluate the diagnostic sensitivity of rickettsial DNA detection in blood and skin samples, we compared results of PCR testing in parallel samples collected from model laboratory animals infected with Rickettsia rickettsii, Rickettsia parkeri and Rickettsia slovaca-like agent at different time points after infection. Skin samples were collected from ears - away from the site of tick placement and without eschars. Overall, testing of skin samples resulted in a higher proportion of positive results than testing of blood samples. Presented data from model animals demonstrates that testing of skin samples from sites of rickettsial proliferation can provide definitive molecular diagnosis of up to 60-70% of tick-borne SFG rickettsial infections during the acute stage of illness. Detection of pathogen DNA in cutaneous samples is a valuable alternative to blood-PCR at least in model animals. Published by Elsevier GmbH.

  3. Brucella abortus is Prevalent in Both Humans and Animals in Bangladesh.

    PubMed

    Rahman, A K M A; Saegerman, C; Berkvens, D; Melzer, F; Neubauer, H; Fretin, D; Abatih, E; Dhand, N; Ward, M P

    2017-08-01

    To determine the role of different Brucella (B.) spp. in Bangladesh, 62 animal samples and 500 human sera were tested. Animal samples from cattle, goats and sheep (including milk, bull semen, vaginal swabs and placentas) were cultured for Brucella spp. Three test-positive human sera and all animal samples were screened by Brucella genus-specific real-time PCR (RT-PCR), and positive samples were then tested by IS711 RT-PCR to detect B. abortus and B. melitensis DNA. Only B. abortus DNA was amplified from 13 human and six animal samples. This is the first report describing B. abortus as the aetiological agent of brucellosis in occupationally exposed humans in Bangladesh. Of note is failure to detect B. melitensis DNA, the species most often associated with human brucellosis worldwide. Further studies are required to explore the occurrence of Brucella melitensis in Bangladesh. © 2017 Blackwell Verlag GmbH.

  4. From milk to diet: feed recognition for milk authenticity.

    PubMed

    Ponzoni, E; Gianì, S; Mastromauro, F; Breviario, D

    2009-11-01

    The presence of plastidial DNA fragments of plant origin in animal milk samples has been confirmed. An experimental plan was arranged with 4 groups of goats, each provided with a different monophytic diet: 3 fresh forages (oats, ryegrass, and X-triticosecale) and one 2-wk-old silage (X-triticosecale). Feed-derived rubisco (ribulose bisphosphate carboxylase, rbcL) DNA fragments were detected in 100% of the analyzed goat milk samples, and the nucleotide sequence of the PCR-amplified fragments was found to be 100% identical to the corresponding fragments amplified from the plant species consumed in the diet. Two additional chloroplast-based molecular markers were used to set up an assay for distinctiveness, conveniently based on a simple PCR. In one case, differences in single nucleotides occurring within the gene encoding for plant maturase K (matK) were exploited. In the other, plant species recognition was based on the difference in the length of the intron present within the transfer RNA leucine (trnL) gene. The presence of plastidial plant DNA, ascertained by the PCR-based amplification of the rbcL fragment, was also assessed in raw cow milk samples collected directly from stock farms or taken from milk sold on the commercial market. In this case, the nucleotide sequence of the amplified DNA fragments reflected the multiple forages present in the diet fed to the animals.

  5. Randomized clinical trial to evaluate the efficacy of a 5-day ceftiofur hydrochloride intramammary treatment on nonsevere gram-negative clinical mastitis.

    PubMed

    Schukken, Y H; Bennett, G J; Zurakowski, M J; Sharkey, H L; Rauch, B J; Thomas, M J; Ceglowski, B; Saltman, R L; Belomestnykh, N; Zadoks, R N

    2011-12-01

    The objective of this study was to evaluate the efficacy of intramammary treatment with ceftiofur hydrochloride of nonsevere, clinical coliform mastitis. One hundred four cases on 5 farms met the enrollment criteria for the study. Escherichia coli was the most common coliform species identified in milk samples from cows with mild to moderate clinical mastitis, followed by Klebsiella spp. and Enterobacter spp. At enrollment, a milk sample from the affected quarter was taken and used for on-farm culture or submitted to the laboratory. For cows in the treatment group, treatment was initiated with ceftiofur hydrochloride via intramammary infusion at 24-h intervals for 5 d according to label standards. Cows in the control group did not receive treatment. Culture results were available on the day after enrollment and only cows with coliform mastitis continued in the treatment and untreated control groups. Bacteriological cure was defined based on 2 posttreatment milk samples. Molecular typing was used for final definition of bacteriological cure. Treatment of nonsevere clinical gram-negative mastitis with ceftiofur hydrochloride resulted in a significant increase in bacteriological cure compared with nontreated controls in animals infected with E. coli or Klebsiella spp. Treated animals clinically improved significantly more compared with control cows. No significant differences were observed between treated and control animals in milk production or linear score before or after clinical mastitis. Treated animals left the study less frequently compared with control animals. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  6. Investigation of pharmaceuticals in processed animal by-products by liquid chromatography coupled to high-resolution mass spectrometry.

    PubMed

    Nácher-Mestre, Jaime; Ibáñez, María; Serrano, Roque; Boix, Clara; Bijlsma, Lubertus; Lunestad, Bjørn Tore; Hannisdal, Rita; Alm, Martin; Hernández, Félix; Berntssen, Marc H G

    2016-07-01

    There is an on-going trend for developing more sustainable salmon feed in which traditionally applied marine feed ingredients are replaced with alternatives. Processed animal products (PAPs) have been re-authorized as novel high quality protein ingredients in 2013. These PAPs may harbor undesirable substances such as pharmaceuticals and metabolites which are not previously associated with salmon farming, but might cause a potential risk for feed and food safety. To control these contaminants, an analytical strategy based on a generic extraction followed by ultra-high performance liquid chromatography coupled to high resolution mass spectrometry (UHPLC-HRMS) using quadrupole time-of-flight mass analyzer (QTOF MS) was applied for wide scope screening. Quality control samples, consisting of PAP commodities spiked at 0.02, 0.1 and 0.2 mg/kg with 150 analytes, were injected in every sample batch to verify the overall method performance. The methodology was applied to 19 commercially available PAP samples from six different types of matrices from the EU animal rendering industry. This strategy allows assessing possible emergent risk exposition of the salmon farming industry to 1005 undesirables, including pharmaceuticals, several dyes and relevant metabolites. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Authentication of animal origin of heparin and low molecular weight heparin including ovine, porcine and bovine species using 1D NMR spectroscopy and chemometric tools.

    PubMed

    Monakhova, Yulia B; Diehl, Bernd W K; Fareed, Jawed

    2018-02-05

    High resolution (600MHz) nuclear magnetic resonance (NMR) spectroscopy is used to distinguish heparin and low-molecular weight heparins (LMWHs) produced from porcine, bovine and ovine mucosal tissues as well as their blends. For multivariate analysis several statistical methods such as principal component analysis (PCA), factor discriminant analysis (FDA), partial least squares - discriminant analysis (PLS-DA), linear discriminant analysis (LDA) were utilized for the modeling of NMR data of more than 100 authentic samples. Heparin and LMWH samples from the independent test set (n=15) were 100% correctly classified according to its animal origin. Moreover, by using 1 H NMR coupled with chemometrics and several batches of bovine heparins from two producers were differentiated. Thus, NMR spectroscopy combined with chemometrics is an efficient tool for simultaneous identification of animal origin and process based manufacturing difference in heparin products. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Nitrogen-to-Protein Conversion Factors for Crop Residues and Animal Manure Common in China.

    PubMed

    Chen, Xueli; Zhao, Guanglu; Zhang, Yang; Han, Lujia; Xiao, Weihua

    2017-10-25

    Accurately determining protein content is essential in exploiting biomass as feed and fuel. A survey of biomass samples in China indicated protein contents from 2.65 to 3.98% for crop residues and from 6.07 to 10.24% for animal manure of dry basis. Conversion factors based on amino acid nitrogen (k A ) ranged from 5.42 to 6.00 for the former and from 4.78 to 5.36 for the latter, indicating that the traditional factor of 6.25 is not suitable for biomass samples. On the other hand, conversion factors from Kjeldahl nitrogen (k P ) ranged from 3.97 to 4.57 and from 2.76 to 4.31 for crop residues and animal manure, respectively. Of note, conversion factors were strongly affected by amino acid composition and levels of nonprotein nitrogen. Thus, k P values of 4.23 for crop residues, 4.11 for livestock manure, and 3.11 for poultry manure are recommended to better estimate protein content from total nitrogen.

  9. Monitoring of small laboratory animal experiments by a designated web-based database.

    PubMed

    Frenzel, T; Grohmann, C; Schumacher, U; Krüll, A

    2015-10-01

    Multiple-parametric small animal experiments require, by their very nature, a sufficient number of animals which may need to be large to obtain statistically significant results.(1) For this reason database-related systems are required to collect the experimental data as well as to support the later (re-) analysis of the information gained during the experiments. In particular, the monitoring of animal welfare is simplified by the inclusion of warning signals (for instance, loss in body weight >20%). Digital patient charts have been developed for human patients but are usually not able to fulfill the specific needs of animal experimentation. To address this problem a unique web-based monitoring system using standard MySQL, PHP, and nginx has been created. PHP was used to create the HTML-based user interface and outputs in a variety of proprietary file formats, namely portable document format (PDF) or spreadsheet files. This article demonstrates its fundamental features and the easy and secure access it offers to the data from any place using a web browser. This information will help other researchers create their own individual databases in a similar way. The use of QR-codes plays an important role for stress-free use of the database. We demonstrate a way to easily identify all animals and samples and data collected during the experiments. Specific ways to record animal irradiations and chemotherapy applications are shown. This new analysis tool allows the effective and detailed analysis of huge amounts of data collected through small animal experiments. It supports proper statistical evaluation of the data and provides excellent retrievable data storage. © The Author(s) 2015.

  10. Buccal swabbing as a noninvasive method to determine bacterial, archaeal, and eukaryotic microbial community structures in the rumen.

    PubMed

    Kittelmann, Sandra; Kirk, Michelle R; Jonker, Arjan; McCulloch, Alan; Janssen, Peter H

    2015-11-01

    Analysis of rumen microbial community structure based on small-subunit rRNA marker genes in metagenomic DNA samples provides important insights into the dominant taxa present in the rumen and allows assessment of community differences between individuals or in response to treatments applied to ruminants. However, natural animal-to-animal variation in rumen microbial community composition can limit the power of a study considerably, especially when only subtle differences are expected between treatment groups. Thus, trials with large numbers of animals may be necessary to overcome this variation. Because ruminants pass large amounts of rumen material to their oral cavities when they chew their cud, oral samples may contain good representations of the rumen microbiota and be useful in lieu of rumen samples to study rumen microbial communities. We compared bacterial, archaeal, and eukaryotic community structures in DNAs extracted from buccal swabs to those in DNAs from samples collected directly from the rumen by use of a stomach tube for sheep on four different diets. After bioinformatic depletion of potential oral taxa from libraries of samples collected via buccal swabs, bacterial communities showed significant clustering by diet (R = 0.37; analysis of similarity [ANOSIM]) rather than by sampling method (R = 0.07). Archaeal, ciliate protozoal, and anaerobic fungal communities also showed significant clustering by diet rather than by sampling method, even without adjustment for potentially orally associated microorganisms. These findings indicate that buccal swabs may in future allow quick and noninvasive sampling for analysis of rumen microbial communities in large numbers of ruminants. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  11. Development of pig welfare assessment protocol integrating animal-, environment-, and management-based measures.

    PubMed

    Renggaman, Anriansyah; Choi, Hong L; Sudiarto, Sartika Ia; Alasaarela, Laura; Nam, Ok S

    2015-01-01

    Due to increased interest in animal welfare, there is now a need for a comprehensive assessment protocol to be used in intensive pig farming systems. There are two current welfare assessment protocols for pigs: Welfare Quality® Assessment Protocols (applicable in the Europe Union), that mostly focuses on animal-based measures, and the Swine Welfare Assurance Program (applicable in the United States), that mostly focuses on management- and environment-based measures. In certain cases, however, animal-based measures might not be adequate for properly assessing pig welfare status. Similarly, welfare assessment that relies only on environment- and management-based measures might not represent the actual welfare status of pigs. Therefore, the objective of this paper was to develop a new welfare protocol by integrating animal-, environment-, and management-based measures. The background for selection of certain welfare criteria and modification of the scoring systems from existing welfare assessment protocols are described. The developed pig welfare assessment protocol consists of 17 criteria that are related to four main principles of welfare (good feeding, good housing, good health, and appropriate behavior). Good feeding, good housing, and good health were assessed using a 3-point scale: 0 (good welfare), 1 (moderate welfare), and 2 (poor welfare). In certain cases, only a 2-point scale was used: 0 (certain condition is present) or 2 (certain condition is absent). Appropriate behavior was assessed by scan sampling of positive and negative social behaviors based on qualitative behavior assessment and human-animal relationship tests. Modification of the body condition score into a 3-point scale revealed pigs with a moderate body condition (score 1). Moreover, additional criteria such as feed quality confirmed that farms had moderate (score 1) or poor feed quality (score 2), especially those farms located in a high relative humidity region. The developed protocol can be utilized to assess welfare status in an intensive pig farming system. Although further improvements are still needed, this study is a first step in developing a pig welfare assessment protocol that combines animal-, environment-, and management-based measures.

  12. A simple device to convert a small-animal PET scanner into a multi-sample tissue and injection syringe counter.

    PubMed

    Green, Michael V; Seidel, Jurgen; Choyke, Peter L; Jagoda, Elaine M

    2017-10-01

    We describe a simple fixture that can be added to the imaging bed of a small-animal PET scanner that allows for automated counting of multiple organ or tissue samples from mouse-sized animals and counting of injection syringes prior to administration of the radiotracer. The combination of imaging and counting capabilities in the same machine offers advantages in certain experimental settings. A polyethylene block of plastic, sculpted to mate with the animal imaging bed of a small-animal PET scanner, is machined to receive twelve 5-ml containers, each capable of holding an entire organ from a mouse-sized animal. In addition, a triangular cross-section slot is machined down the centerline of the block to secure injection syringes from 1-ml to 3-ml in size. The sample holder is scanned in PET whole-body mode to image all samples or in one bed position to image a filled injection syringe. Total radioactivity in each sample or syringe is determined from the reconstructed images of these objects using volume re-projection of the coronal images and a single region-of-interest for each. We tested the accuracy of this method by comparing PET estimates of sample and syringe activity with well counter and dose calibrator estimates of these same activities. PET and well counting of the same samples gave near identical results (in MBq, R 2 =0.99, slope=0.99, intercept=0.00-MBq). PET syringe and dose calibrator measurements of syringe activity in MBq were also similar (R 2 =0.99, slope=0.99, intercept=- 0.22-MBq). A small-animal PET scanner can be easily converted into a multi-sample and syringe counting device by the addition of a sample block constructed for that purpose. This capability, combined with live animal imaging, can improve efficiency and flexibility in certain experimental settings. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Are most samples of animals systematically biased? Consistent individual trait differences bias samples despite random sampling.

    PubMed

    Biro, Peter A

    2013-02-01

    Sampling animals from the wild for study is something nearly every biologist has done, but despite our best efforts to obtain random samples of animals, 'hidden' trait biases may still exist. For example, consistent behavioral traits can affect trappability/catchability, independent of obvious factors such as size and gender, and these traits are often correlated with other repeatable physiological and/or life history traits. If so, systematic sampling bias may exist for any of these traits. The extent to which this is a problem, of course, depends on the magnitude of bias, which is presently unknown because the underlying trait distributions in populations are usually unknown, or unknowable. Indeed, our present knowledge about sampling bias comes from samples (not complete population censuses), which can possess bias to begin with. I had the unique opportunity to create naturalized populations of fish by seeding each of four small fishless lakes with equal densities of slow-, intermediate-, and fast-growing fish. Using sampling methods that are not size-selective, I observed that fast-growing fish were up to two-times more likely to be sampled than slower-growing fish. This indicates substantial and systematic bias with respect to an important life history trait (growth rate). If correlations between behavioral, physiological and life-history traits are as widespread as the literature suggests, then many animal samples may be systematically biased with respect to these traits (e.g., when collecting animals for laboratory use), and affect our inferences about population structure and abundance. I conclude with a discussion on ways to minimize sampling bias for particular physiological/behavioral/life-history types within animal populations.

  14. Factors influencing the demand of the service of community based animal health care in Zimbabwe.

    PubMed

    Mutambara, J; Dube, I; Matangi, E; Majeke, F

    2013-11-01

    This study was done to find out about animal health service providers and factors that determined demand for community based veterinary service delivery in smallholder sector of Zimbabwe. Focus group discussions and a questionnaire was used to collect data on veterinary services providers and socio-economic factors related to animal health from a sample (N=333) smallholder livestock farmers from Gutu district of Masvingo province in Zimbabwe. Analytical techniques used were descriptive statistics, K-mean cluster analysis and Tobit regression model. Results showed that the majority of farmers (45%) obtained services from both Community Based Animal Health Workers (CBAHWs) and Department of Veterinary Service (DVS), 25% DVS only, 20% used CBAHWs while 10% did not seek any services. Further analysis showed that distance to CBAHW, distance to AHMC and employment status were significantly related to demand for CBAHWs with coefficients of -1.5, 0.7 and -10.3, respectively. The study thus concluded that CBAHW is an alternative animal health service delivery approach already practiced in smallholder farming sectors of Zimbabwe. Socio-economic factors significantly influenced the demand for CBAHW services. Given limited resources by state sponsored veterinary services, it is recommended that the CBAHWs approach should be encouraged as supplementary service provider especially in areas further DVS. These community organizations can be empowered by the state to deliver more improved services based on hygiene and modern science at a relatively low cost to farmers. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Effectiveness of brain-based learning and animated cartoons for enhancing healthy habits among school children in Khon Kaen, Thailand.

    PubMed

    Banchonhattakit, Pannee; Duangsong, Rujira; Muangsom, Niramon; Kamsong, Theppamon; Phangwan, Krittiya

    2015-03-01

    The objective of this study was to investigate the effectiveness of brain-based learning (BBL) and animated cartoons on video compact discs (VCDs) in enhancing the healthy habits of school children. A representative sample of 1085 school children in the first through the third grades at 16 schools was selected by multistage random sampling. Knowledge of healthy habits and self-reported adoption of practices were assessed by a questionnaire. BBL and VCD, either combined or as single-intervention techniques, led to improved knowledge and practice of healthy behavior, whereas conventional teaching did not. As a single-intervention technique, BBL on its own led to a greater improvement in healthy practices than VCD, but the addition of BBL to VCD made no difference, and there was no difference between BBL and VCD in terms of improvements in knowledge. In conclusion, both BBL and VCD are effective, but VCD requires fewer resources. Recommendations are made for further research. © 2012 APJPH.

  16. First-time detection of Mycobacterium bovis in livestock tissues and milk in the West Bank, Palestinian Territories.

    PubMed

    Ereqat, Suheir; Nasereddin, Abedelmajeed; Levine, Hagai; Azmi, Kifaya; Al-Jawabreh, Amer; Greenblatt, Charles L; Abdeen, Ziad; Bar-Gal, Gila Kahila

    2013-01-01

    Bovine tuberculosis, bTB, is classified by the WHO as one of the seven neglected zoonontic diseases that cause animal health problems and has high potential to infect humans. In the West Bank, bTB was not studied among animals and the prevalence of human tuberculosis caused by M. bovis is unknown. Therefore, the aim of this study was to estimate the prevalence of bTB among cattle and goats and identify the molecular characteristics of bTB in our area. A total of 208 tissue samples, representing 104 animals, and 150 raw milk samples, obtained from cows and goats were examined for the presence of mycobacteria. The tissue samples were collected during routine meat inspection from the Jericho abattoir. DNA was extracted from all samples, milk and tissue biopsies (n = 358), and screened for presence of TB DNA by amplifying a 123-bp segment of the insertion sequence IS6110. Eight out of 254 animals (3.1%) were found to be TB positive based on the IS6110-PCR. Identification of M. bovis among the positive TB samples was carried out via real time PCR followed by high resolution melt curve analysis, targeting the A/G transition along the oxyR gene. Spoligotyping analysis revealed a new genotype of M. bovis that was revealed from one tissue sample. Detection of M. bovis in tissue and milk of livestock suggests that apparently healthy cattle and goats are a potential source of infection of bTB and may pose a risk to public health. Hence, appropriate measures including meat inspection at abattoirs in the region are required together with promotion of a health campaign emphasizing the importance of drinking pasteurized milk. In addition, further studies are essential at the farm level to determine the exact prevalence of bTB in goats and cattle herds in the West Bank and Israel.

  17. First-Time Detection of Mycobacterium bovis in Livestock Tissues and Milk in the West Bank, Palestinian Territories

    PubMed Central

    Ereqat, Suheir; Nasereddin, Abedelmajeed; Levine, Hagai; Azmi, Kifaya; Al-Jawabreh, Amer; Greenblatt, Charles L.; Abdeen, Ziad; Bar-Gal, Gila Kahila

    2013-01-01

    Background Bovine tuberculosis, bTB, is classified by the WHO as one of the seven neglected zoonontic diseases that cause animal health problems and has high potential to infect humans. In the West Bank, bTB was not studied among animals and the prevalence of human tuberculosis caused by M. bovis is unknown. Therefore, the aim of this study was to estimate the prevalence of bTB among cattle and goats and identify the molecular characteristics of bTB in our area. Methodology/principal findings A total of 208 tissue samples, representing 104 animals, and 150 raw milk samples, obtained from cows and goats were examined for the presence of mycobacteria. The tissue samples were collected during routine meat inspection from the Jericho abattoir. DNA was extracted from all samples, milk and tissue biopsies (n = 358), and screened for presence of TB DNA by amplifying a 123-bp segment of the insertion sequence IS6110. Eight out of 254 animals (3.1%) were found to be TB positive based on the IS6110-PCR. Identification of M. bovis among the positive TB samples was carried out via real time PCR followed by high resolution melt curve analysis, targeting the A/G transition along the oxyR gene. Spoligotyping analysis revealed a new genotype of M. bovis that was revealed from one tissue sample. Significance Detection of M. bovis in tissue and milk of livestock suggests that apparently healthy cattle and goats are a potential source of infection of bTB and may pose a risk to public health. Hence, appropriate measures including meat inspection at abattoirs in the region are required together with promotion of a health campaign emphasizing the importance of drinking pasteurized milk. In addition, further studies are essential at the farm level to determine the exact prevalence of bTB in goats and cattle herds in the West Bank and Israel. PMID:24069475

  18. Teaching habitat and animal classification to fourth graders using an engineering-design model

    NASA Astrophysics Data System (ADS)

    Marulcu, Ismail

    2014-05-01

    Background: The motivation for this work is built upon the premise that there is a need for research-based materials for design-based science instruction. In this paper, a small portion of our work investigating the impact of a LEGOTM engineering unit on fourth grade students' preconceptions and understanding of animals is presented. Purpose: The driving questions for our work are: (1) What is the impact of an engineering-design-based curricular module on students' understanding of habitat and animal classification? (2) What are students' misconceptions regarding animal classification and habitat? Sample: The study was conducted in an inner-city K-8 school in the northeastern region of the United States. There were two fourth grade classrooms in the school. The first classroom included seven girls and nine boys, whereas the other classroom included eight girls and eight boys. All fourth grade students participated in the study. Design and methods: In answering the research questions mixed-method approaches are used. Data collection methods included pre- and post-tests, pre- and post-interviews, student journals, and classroom observations. Identical pre- and post-tests were administered to measure students' understanding of animals. They included four multiple-choice and six open-ended questions. Identical pre- and post-interviews were administered to explore students' in-depth understanding of animals. Results: Our results show that students significantly increased their performance after instruction on both the multiple-choice questions (t = -3.586, p = .001) and the open-ended questions (t = -5.04, p = .000). They performed better on the post interviews as well. Also, it is found that design-based instruction helped students comprehend core concepts of a life science subject, animals. Conclusions: Based on these results, the main argument of the study is that engineering design is a useful framework for teaching not only physical science-related subjects, but also life science subjects in elementary science classrooms.

  19. OCCURRENCE OF INTRINSIC VANCOMYCIN RESISTANT ENTEROCOCCI IN ANIMAL FECES

    EPA Science Inventory

    A survey was conducted to determine the occurrence of vancomycin resistant enterococci (VRE) in animal and human fecal samples. Fecal samples from 14 animal species and humans were analyzed by quantitative culture for enterococci and VRE. Over 800 VRE isolates were characterize...

  20. Animal-Assisted Interventions in the Classroom-A Systematic Review.

    PubMed

    Brelsford, Victoria L; Meints, Kerstin; Gee, Nancy R; Pfeffer, Karen

    2017-06-22

    The inclusion of animals in educational practice is becoming increasingly popular, but it is unclear how solid the evidence for this type of intervention is. The aim of this systematic review is to scrutinise the empirical research literature relating to animal-assisted interventions conducted in educational settings. The review included 25 papers; 21 from peer-reviewed journals and 4 obtained using grey literature databases. Most studies reported significant benefits of animal-assisted interventions in the school setting. Despite this, studies vary greatly in methods and design, in intervention types, measures, and sample sizes, and in the length of time exposed to an animal. Furthermore, a worrying lack of reference to risk assessment and animal welfare must be highlighted. Taken together, the results of this review show promising findings and emerging evidence suggestive of potential benefits related to animals in school settings. The review also indicates the need for a larger and more robust evidence base driven by thorough and strict protocols. The review further emphasises the need for safeguarding for all involved-welfare and safety are paramount.

  1. Retrospective investigation of chronic wasting disease of cervids at the Toronto Zoo, 1973–2003

    PubMed Central

    Dubé, Caroline; Mehren, Kay G.; Barker, Ian K.; Peart, Brian L.; Balachandran, Aru

    2006-01-01

    The occurrence of chronic wasting disease (CWD) at the Toronto Zoo was investigated retrospectively, based on an examination of management, animal health, and postmortem records, and immunohistochemical studies. Records of animal movements, clinical signs, and postmortem findings were examined for all cervids 1973–2003. All available samples of fixed, wax-embedded lymphoid or central nervous system tissue from cervids that died at the Toronto Zoo from 1973 to 2003, > 12 months of age, were tested, using prion protein immunostaining. Chronic wasting disease prion antigen was detected in 8 of 105 animals tested: 7 mule deer and 1 black-tailed deer. The most likely method of introduction was the importation of CWD-infected animals from a zoo in the United States. Animal-to-animal contact and environmental contamination were the most likely methods of spread of CWD at the zoo. No mule deer left the Toronto Zoo site, and the last animal with CWD died in 1981. Historic findings and ongoing testing of cervids indicate that the Toronto Zoo collection has very low risk of currently being infected with CWD. PMID:17217088

  2. Survey on Animal Welfare in Nine Hundred and Forty Three Italian Dairy Farms

    PubMed Central

    Pietra, Marco; Giacometti, Federica; Mazzi, Antonella; Scacco, Gianluca; Serraino, Andrea; Scagliarini, Lorenzo

    2016-01-01

    The final results of a survey on welfare of dairy cows in 7 Italian Regions are presented. The study has been performed on 943 farms in southern and central Italy to highlight critical and strong points concerning animal welfare in dairy systems, by using direct and indirect criteria. To assess animal welfare, a checklist based on 303 parameters has been used; indirect criteria have been organised in 5 general areas concerning Farm management, Farming and housing systems, Environment, Feeding, Health and hygiene; other resource-based criteria were considered in 5 specific areas for the different productive categories (lactating cows, dry cows, pregnant heifers, cows comeback, calves up to 8 weeks and calves between 8 weeks and 6 months); finally, an Indicators section focused on animal based criteria. Parameters have been valued as conforming or not conforming on the basis of the current lesgislation on animal welfare, and in the other cases by the use of a semi-quantitative scale such as poor, satisfactory, good or very good referring to scientific literature and reports by the Animal Health and Animal Welfare panel of the European Food Safety Authority. Among the 249 examined parameters (54 criteria have been valued as descriptive), 15 showed a failure prevalence inferior to 1%; for the remaining parameters, the overall non-compliance prevalence on the whole sample ranged from a maximum of 67% to a minimum of 2%, showing an inverse proportionality correlation with the herd size. One hundred and ten parameters were judged as poor (96) or not in compliance with the rules in force (14) in more than 10% of the examined herds. The most common non-compliance aspects detected in the different areas concern calves management, staff training and prophylaxis programmes; staff training levels were inversely related to failure prevalences in almost all areas. The combination of direct and indirect criteria has allowed to fully embrace recommendations on the use of animal based measures for the assessment of animal welfare, as accepted into the strategic Plan for the EU animal welfare for 2012-2015. PMID:27800437

  3. IDENTIFICATION OF BACTERIA IN BLOOD CULTURES FROM CLINICALLY ILL CAPTIVE ANTILLEAN MANATEES (TRICHECHUS MANATUS MANATUS).

    PubMed

    Silva, Mariana C O; Attademo, Fernanda F L; Freire, Augusto C B; Sousa, Glaucia P; Luna, Fábia O; Lima, Débora C V; Mota, Rinaldo A; Mendes, Emiko S; Silva, Jean C R

    2017-03-01

    Between September 2001 and March 2013, 62 bacterial cultures (37 aerobic and 25 anaerobic) were performed on 37 blood samples from 23 Antillean manatees ( Trichechus manatus manatus) that were kept in captivity at the Brazilian National Center for Research and Conservation of Aquatic Mammals (CMA) in Pernambuco (CMA-PE) and Alagoas (CMA-AL), Brazil. All of the animals sampled exhibited clinical signs at the time of sampling including abscesses (n = 8), debilitation and anorexia (n = 22), and profound lethargy-moribundity (n = 7). The 4 animals with profound lethargy-moribundity died shortly after sampling of unknown causes. Bacteria were isolated from 15/37 (40.5%) and aerobic blood cultures from 13/23 animals (56.5%). None of the anaerobic cultures were positive. Aeromonas caviae , Aeromonas hydrophila , Aeromonas sp., Escherichia coli , Leclercia adecarboxylata , Pantoea agglomerans , Pseudomonas aeruginosa , Pseudomonas stutzeri , Pseudomonas sp., Sphingomonas paucimobilis , coagulase-negative Staphylococcus, and Staphylococcus epidermidis were each found in only one animal; Staphylococcus spp. was found in two; and Vibrio fluvialis in four. Thirteen samples had only one bacteria isolated, one sample had two bacteria, and one sample had three bacteria isolated. Regarding sex, age group, and origin among the manatees examined, 54.5% (6/11) of the females, 58.3% (7/12) of the males, 40% (2/5) of the calves, 66.7% (8/12) of the juveniles, 50% (3/6) of the adults, 55.5% (10/18) at CMA-PE, and 60% (3/5) at CMA-AL were found to be positive for bacterial growth during at least one sampling time. All Antillean manatees were clinically ill. Regarding clinical signs, bacteria were found in 50% (11/22) of blood samples of the animals showing debilitation and anorexia, 1 of 8 (12.5%) of blood samples of the animals showing abscesses, and 3 of 7 (42.9%) of blood samples of the animals showing profound lethargy-moribundity.

  4. Estimating rates of local species extinction, colonization and turnover in animal communities

    USGS Publications Warehouse

    Nichols, James D.; Boulinier, T.; Hines, J.E.; Pollock, K.H.; Sauer, J.R.

    1998-01-01

    Species richness has been identified as a useful state variable for conservation and management purposes. Changes in richness over time provide a basis for predicting and evaluating community responses to management, to natural disturbance, and to changes in factors such as community composition (e.g., the removal of a keystone species). Probabilistic capture-recapture models have been used recently to estimate species richness from species count and presence-absence data. These models do not require the common assumption that all species are detected in sampling efforts. We extend this approach to the development of estimators useful for studying the vital rates responsible for changes in animal communities over time; rates of local species extinction, turnover, and colonization. Our approach to estimation is based on capture-recapture models for closed animal populations that permit heterogeneity in detection probabilities among the different species in the sampled community. We have developed a computer program, COMDYN, to compute many of these estimators and associated bootstrap variances. Analyses using data from the North American Breeding Bird Survey (BBS) suggested that the estimators performed reasonably well. We recommend estimators based on probabilistic modeling for future work on community responses to management efforts as well as on basic questions about community dynamics.

  5. Limitations to mapping habitat-use areas in changing landscapes using the Mahalanobis distance statistic

    USGS Publications Warehouse

    Knick, Steven T.; Rotenberry, J.T.

    1998-01-01

    We tested the potential of a GIS mapping technique, using a resource selection model developed for black-tailed jackrabbits (Lepus californicus) and based on the Mahalanobis distance statistic, to track changes in shrubsteppe habitats in southwestern Idaho. If successful, the technique could be used to predict animal use areas, or those undergoing change, in different regions from the same selection function and variables without additional sampling. We determined the multivariate mean vector of 7 GIS variables that described habitats used by jackrabbits. We then ranked the similarity of all cells in the GIS coverage from their Mahalanobis distance to the mean habitat vector. The resulting map accurately depicted areas where we sighted jackrabbits on verification surveys. We then simulated an increase in shrublands (which are important habitats). Contrary to expectation, the new configurations were classified as lower similarity relative to the original mean habitat vector. Because the selection function is based on a unimodal mean, any deviation, even if biologically positive, creates larger Malanobis distances and lower similarity values. We recommend the Mahalanobis distance technique for mapping animal use areas when animals are distributed optimally, the landscape is well-sampled to determine the mean habitat vector, and distributions of the habitat variables does not change.

  6. Microscopic and Molecular Detection of Theileria (Babesia) Equi Infection in Equids of Kurdistan Province, Iran.

    PubMed

    Habibi, Gholamreza; Esmaeilnia, Kasra; Hablolvarid, Mohammad Hasan; Afshari, Asghar; Zamen, Mohsen; Bozorgi, Soghra

    2016-01-01

    Equine piroplasmosis (EP) is the cause of persistent tick-borne infection with no symptoms, but the most important problem of EP is due to the persistent carrier state. Carrier animals to Babesia (Theileria) equi (Laveran 1901) and B. caballi (Nuttall, 1910) infestation could be identified by extremely sensitive PCR-based method. The purpose of this study was to identify the causative agents of equine piroplasmosis based on molecular and microscopic assays in equids from Kurdistan Province, Iran. Thirty one horse and mule blood samples were used with history of living in Kurdistan Province of Iran. The blood specimens were utilized for T. equi and B. caballi DNA identification by PCR and Giemsa stained smears for microscopic observation. The results clearly showed the presence of B. (Theileria) equi DNA in 30 of 31 blood samples (96.77%), but the microscopic examination revealed the 3 of 31 positive Babesia like organisms in the red blood cells (9.67%). The obtained results demonstrated the presence of hidden B. (Theileria) equi infection in horses with previous habitance in Kurdistan Province of Iran. The carrier animals became a main source of infection and can transmit the disease. Therefore, hidden infection might be considered as a health threatening and limiting factor in animals used in therapeutic antisera research and production centers.

  7. Point prevalence survey for tick-borne pathogens in military working dogs, shelter animals, and pet populations in northern Colombia.

    PubMed

    McCown, M E; Alleman, A; Sayler, K A; Chandrashekar, R; Thatcher, B; Tyrrell, P; Stillman, B; Beall, M; Barbet, A F

    2014-01-01

    Based on the high tick-borne pathogen results from a 2011 surveillance study in three Colombian cities, an in-depth point prevalence survey was conducted to determine the seroprevalence of tick-borne pathogens at a specific point in time in 70 working dogs, 101 shelter dogs, and 47 client-owned dogs in Barranquilla, Colombia. Of the 218 serum samples, 163 (74%) were positive for Ehrlichia canis and 116 (53%) for Anaplasma platys. Exposure to tick-borne pathogens was highest in shelter and working dogs where more than 90% of the samples were seropositive or positive on polymerase chain reaction for one or more organisms as compared to 51% in client-owned animals. Surveillance for exposure to tick-borne pathogens provides vital information necessary to protect and conserve the health of local humans and animals, deployed military service members, and working dogs in various parts of the world. This study and resultant data demonstrate the value of following a broad-based surveillance study with a more specific, focused analysis in an area of concern. This area?s high levels of exposure warrant emphasis by medical planners and advisors on precautionary measures for military dogs, Special Operations Forces personnel, and the local public. 2014.

  8. Prevalence, serovars, phage types, and antibiotic susceptibilities of Salmonella strains isolated from animals in the United Arab Emirates from 1996 to 2009.

    PubMed

    Münch, Sebastian; Braun, Peggy; Wernery, Ulrich; Kinne, Jörg; Pees, Michael; Flieger, Antje; Tietze, Erhard; Rabsch, Wolfgang

    2012-10-01

    The aim of this study was to give some insights into the prevalence, serovars, phage types, and antibiotic resistances of Salmonella from animal origin in the United Arab Emirates. Data on diagnostic samples from animals (n = 20,871) examined for Salmonella between 1996 and 2009 were extracted from the databases of the Central Veterinary Research Laboratory in Dubai and from typed strains (n = 1052) from the Robert Koch Institute, Wernigerode Branch in Germany and analyzed for general and animal-specific trends. Salmonella was isolated from 1,928 (9 %) of the 20,871 samples examined. Among the 1,052 typed strains, most were from camels (n = 232), falcons (n = 166), bustards (n = 101), antelopes (n = 66), and horses (n = 63). The predominant serovars were Salmonella Typhimurium (25 %), Salmonella Kentucky (8 %), followed by Salmonella Frintrop (7 %), and Salmonella Hindmarsh (5 %). When analyzed by animal species, the most frequent serovars in camels were Salmonella Frintrop (28 %) and Salmonella Hindmarsh (21 %), in falcons Salmonella Typhimurium (32 %), in bustards Salmonella Kentucky (19 %), in antelopes Salmonella Typhimurium (9 %), and in horses Salmonella Typhimurium (17 %) and S. Kentucky (16 %). Resistance of all typed Salmonella strains (n = 1052) was most often seen to tetracycline (23 %), streptomycin (22 %), nalidixic acid (18 %), and ampicillin (15 %). These data show trends in the epidemiology of Salmonella in different animal species which can be used as a base for future prevention, control, and therapy strategies.

  9. Animal Allergens and Their Presence in the Environment

    PubMed Central

    Zahradnik, Eva; Raulf, Monika

    2014-01-01

    Exposure to animal allergens is a major risk factor for sensitization and allergic diseases. Besides mites and cockroaches, the most important animal allergens are derived from mammals. Cat and dog allergies affect the general population; whereas, allergies to rodents or cattle is an occupational problem. Exposure to animal allergens is not limited to direct contact to animals. Based on their aerodynamic properties, mammalian allergens easily become airborne, attach to clothing and hair, and can be spread from one environment to another. For example, the major cat allergen Fel d 1 was frequently found in homes without pets and in public buildings, including schools, day-care centers, and hospitals. Allergen concentrations in a particular environment showed high variability depending on numerous factors. Assessment of allergen exposure levels is a stepwise process that involves dust collection, allergen quantification, and data analysis. Whereas a number of different dust sampling strategies are used, ELISA assays have prevailed in the last years as the standard technique for quantification of allergen concentrations. This review focuses on allergens arising from domestic, farm, and laboratory animals and describes the ubiquity of mammalian allergens in the human environment. It includes an overview of exposure assessment studies carried out in different indoor settings (homes, schools, workplaces) using numerous sampling and analytical methods and summarizes significant factors influencing exposure levels. However, methodological differences among studies have contributed to the variability of the findings and make comparisons between studies difficult. Therefore, a general standardization of methods is needed and recommended. PMID:24624129

  10. SNP Data Quality Control in a National Beef and Dairy Cattle System and Highly Accurate SNP Based Parentage Verification and Identification

    PubMed Central

    McClure, Matthew C.; McCarthy, John; Flynn, Paul; McClure, Jennifer C.; Dair, Emma; O'Connell, D. K.; Kearney, John F.

    2018-01-01

    A major use of genetic data is parentage verification and identification as inaccurate pedigrees negatively affect genetic gain. Since 2012 the international standard for single nucleotide polymorphism (SNP) verification in Bos taurus cattle has been the ISAG SNP panels. While these ISAG panels provide an increased level of parentage accuracy over microsatellite markers (MS), they can validate the wrong parent at ≤1% misconcordance rate levels, indicating that more SNP are needed if a more accurate pedigree is required. With rapidly increasing numbers of cattle being genotyped in Ireland that represent 61 B. taurus breeds from a wide range of farm types: beef/dairy, AI/pedigree/commercial, purebred/crossbred, and large to small herd size the Irish Cattle Breeding Federation (ICBF) analyzed different SNP densities to determine that at a minimum ≥500 SNP are needed to consistently predict only one set of parents at a ≤1% misconcordance rate. For parentage validation and prediction ICBF uses 800 SNP (ICBF800) selected based on SNP clustering quality, ISAG200 inclusion, call rate (CR), and minor allele frequency (MAF) in the Irish cattle population. Large datasets require sample and SNP quality control (QC). Most publications only deal with SNP QC via CR, MAF, parent-progeny conflicts, and Hardy-Weinberg deviation, but not sample QC. We report here parentage, SNP QC, and a genomic sample QC pipelines to deal with the unique challenges of >1 million genotypes from a national herd such as SNP genotype errors from mis-tagging of animals, lab errors, farm errors, and multiple other issues that can arise. We divide the pipeline into two parts: a Genotype QC and an Animal QC pipeline. The Genotype QC identifies samples with low call rate, missing or mixed genotype classes (no BB genotype or ABTG alleles present), and low genotype frequencies. The Animal QC handles situations where the genotype might not belong to the listed individual by identifying: >1 non-matching genotypes per animal, SNP duplicates, sex and breed prediction mismatches, parentage and progeny validation results, and other situations. The Animal QC pipeline make use of ICBF800 SNP set where appropriate to identify errors in a computationally efficient yet still highly accurate method. PMID:29599798

  11. Ante mortem identification of BSE from serum using infrared spectroscopy

    NASA Astrophysics Data System (ADS)

    Schmitt, Jürgen; Lasch, Peter; Beekes, Michael; Udelhoven, Thomas; Eiden, Michael; Fabian, Heinz; Petrich, Wolfgang H.; Naumann, Dieter

    2004-07-01

    In our former studies a diagnostic approach for the detection of transmissible spongiform encephalopaties (TSE) based on FT-IR spectroscopy in combination with artificial neural networks was described, based on a controlled animal study with terminally ill Syrian hamsters and control animals. As a consequence of the bovine spongiform encephalopathy (BSE) crisis in Europe, the development of a disgnostic ante mortem test for cattle has become a matter of great scientific importance and public interest. Since 1986 more than 180,000 clinical cases of BSE have been observed in the UK alone. Most of these cases were confirmed by post mortem examination of brain tissue. However, BSE-related risk assessment and risk-management would greatly benefit from ante mortem testing on living animals. For example, a serum-based test could allow for screening of the cattle population, thus, even a BSE eradication program would be conceivable. Here we report on a novel method for ante mortem BSE testing, which combines infrared spectroscopy of serum samples with multivariate pattern recognition analysis. A classification algorithm was trained using infrared spectra of sera from more than 800 animals from a field study (including BSE positive, healthy controls and animals suffering from viral or bacterial infections). In two validation studies sensitivities of 85% and 87% and specificities of 84% and 91% were achieved, respectively. The combination of classification algorithms increased sensitivity and specificity to 96% and 92%, respectively.

  12. Presumptive Sources of Fecal Contamination in Four Tributaries to the New River Gorge National River, West Virginia, 2004

    USGS Publications Warehouse

    Mathes, Melvin V.; O'Brien, Tara L.; Strickler, Kriston M.; Hardy, Joshua J.; Schill, William B.; Lukasik, Jerzy; Scott, Troy M.; Bailey, David E.; Fenger, Terry L.

    2007-01-01

    Several methods were used to determine the sources of fecal contamination in water samples collected during September and October 2004 from four tributaries to the New River Gorge National River -- Arbuckle Creek, Dunloup Creek, Keeney Creek, and Wolf Creek. All four tributaries historically have had elevated levels of fecal coliform bacteria. The source-tracking methods used yielded various results, possibly because one or more methods failed. Sourcing methods used in this study included the detection of several human-specific and animal-specific biological or molecular markers, and library-dependent pulsed-field gel electrophoresis analysis that attempted to associate Escherichia coli bacteria obtained from water samples with animal sources by matching DNA-fragment banding patterns. Evaluation of the results of quality-control analysis indicated that pulsed-field gel electrophoresis analysis was unable to identify known-source bacteria isolates. Increasing the size of the known-source library did not improve the results for quality-control samples. A number of emerging methods, using markers in Enterococcus, human urine, Bacteroidetes, and host mitochondrial DNA, demonstrated some potential in associating fecal contamination with human or animal sources in a limited analysis of quality-control samples. All four of the human-specific markers were detected in water samples from Keeney Creek, a watershed with no centralized municipal wastewater-treatment facilities, thus indicating human sources of fecal contamination. The human-specific Bacteroidetes and host mitochondrial DNA markers were detected in water samples from Dunloup Creek, Wolf Creek, and to a lesser degree Arbuckle Creek. Results of analysis for wastewater compounds indicate that the September 27 sample from Arbuckle Creek contained numerous human tracer compounds likely from sewage. Dog, horse, chicken, and pig host mitochondrial DNA were detected in some of the water samples with the exception of the October 5 sample from Dunloup Creek. Cow, white-tailed deer, and Canada goose DNA were not detected in any of the samples collected from the four tributaries, despite the presence of these animals in the watersheds. Future studies with more rigorous quality-control analyses are needed to investigate the potential applicability and use of these emerging methods. Because many of the detections for the various methods could vary over time and with flow conditions, repeated sampling during both base flow and storm events would be necessary to more definitively determine the sources of fecal contamination for each watershed.

  13. Logical fallacies in animal model research.

    PubMed

    Sjoberg, Espen A

    2017-02-15

    Animal models of human behavioural deficits involve conducting experiments on animals with the hope of gaining new knowledge that can be applied to humans. This paper aims to address risks, biases, and fallacies associated with drawing conclusions when conducting experiments on animals, with focus on animal models of mental illness. Researchers using animal models are susceptible to a fallacy known as false analogy, where inferences based on assumptions of similarities between animals and humans can potentially lead to an incorrect conclusion. There is also a risk of false positive results when evaluating the validity of a putative animal model, particularly if the experiment is not conducted double-blind. It is further argued that animal model experiments are reconstructions of human experiments, and not replications per se, because the animals cannot follow instructions. This leads to an experimental setup that is altered to accommodate the animals, and typically involves a smaller sample size than a human experiment. Researchers on animal models of human behaviour should increase focus on mechanistic validity in order to ensure that the underlying causal mechanisms driving the behaviour are the same, as relying on face validity makes the model susceptible to logical fallacies and a higher risk of Type 1 errors. We discuss measures to reduce bias and risk of making logical fallacies in animal research, and provide a guideline that researchers can follow to increase the rigour of their experiments.

  14. DETECTION OF INTRINSIC VANCOMYCIN RESISTANT ENTEROCOCCI IN ANIMAL AND HUMAN FECES

    EPA Science Inventory

    A survey was conducted to determine the occurrence of vancomycin resistant enterococci (VRE) in animal and human fecal samples. Fecal samples from 14 animal species and humans were analyzed by quantitative culture for enterococci and VRE. Over 800 VRE isolates were characterize...

  15. Detection of kanamycin and gentamicin residues in animal-derived food using IgY antibody based ic-ELISA and FPIA.

    PubMed

    Li, Cui; Zhang, Yaoyao; Eremin, Sergei A; Yakup, Omar; Yao, Gang; Zhang, Xiaoying

    2017-07-15

    Our aim in this study is to show that IgY antibody based immunoassays could be used to detect antibiotic residues in animal-derived food. Briefly, full antigens of gentamicin (Gent) and kanamycin (Kana) were used to immunize the laying chickens to prepare IgY antibodies. Then, these antibodies were evaluated by FPIA and ic-ELISA to detect Gent/Kana in animal-derived samples. The IC 50 of FPIA and ic-ELISA based anti-Gent IgY were 7.70±0.6μg/mL and 0.32±0.06μg/mL, respectively. The IC 50 of FPIA and ic-ELISA based anti-Kana IgY were 7.97±0.9μg/mL and 0.15±0.01μg/mL. The limits of detection (LOD, IC 10 ) for FPIA based anti-Gent/Kana IgY were 0.17 and 0.007μg/mL, respectively. The LOD for ic-ELISA were both 0.001μg/mL. These results indicated that the ic-ELISA might more suitable for antibiotic residues detection than FPIA. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Investigation of irradiated rats DNA in the presence of Cu(II) chelates of amino acids Schiff bases.

    PubMed

    Karapetyan, N H; Torosyan, A L; Malakyan, M; Bajinyan, S A; Haroutiunian, S G

    2016-01-01

    The new synthesized Cu(II) chelates of amino acids Schiff bases were studied as a potential radioprotectors. Male albino rats of Wistar strain were exposed to X-ray whole-body irradiation at 4.8 Gy. This dose caused 30% mortality of the animals (LD30). The survival of animals exposed to radiation after preliminary administration of 10 mg/kg Cu(II)(Nicotinyl-L-Tyrosinate)2 or Cu(II)(Nicotinyl-L-Tryptophanate)2 prior to irradiation was registered about 80 and 100% correspondingly. Using spectrophotometric melting and agarose gel electrophoresis methods, the differences between the DNA isolated from irradiated rats and rats pretreated with Cu(II) chelates were studied. The fragments of DNA with different breaks were revealed in DNA samples isolated from irradiated animals. While, the repair of the DNA structure was observed for animals pretreated with the Cu(II) chelates. The results suggested that pretreatment of the irradiated rats with Cu(II)(Nicotinyl-L-Tyrosinate)2 and Cu(II)(Nicotinyl-L-Tryptophanate)2 compounds improves the liver DNA characteristics.

  17. Quantification of Shiga Toxin-Converting Bacteriophages in Wastewater and in Fecal Samples by Real-Time Quantitative PCR ▿

    PubMed Central

    Imamovic, Lejla; Ballesté, Elisenda; Jofre, Juan; Muniesa, Maite

    2010-01-01

    Shiga toxin-converting bacteriophages (Stx phages) are involved in the pathogenicity of some enteric bacteria, such as Escherichia coli O157:H7. Stx phages are released from their bacterial hosts after lytic induction and remain free in the environment. Samples were analyzed for the presence of free Stx phages by an experimental approach based on the use of real-time quantitative PCR (qPCR), which enables stx to be detected in the DNA from the viral fraction of each sample. A total of 150 samples, including urban raw sewage samples, wastewater samples with fecal contamination from cattle, pigs, and poultry, and fecal samples from humans and diverse animals, were used in this study. Stx phages were detected in 70.0% of urban sewage samples (10 to 103 gene copies [GC] per ml) and in 94.0% of animal wastewater samples of several origins (10 to 1010 GC per ml). Eighty-nine percent of cattle fecal samples were positive for Stx phages (10 to 105 GC per g of sample), as were 31.8% of other fecal samples of various origins (10 to 104 GC per g of sample). The stx2 genes and stx2 variants were detected in the viral fraction of some of the samples after sequencing of stx2 fragments amplified by conventional PCR. The occurrence and abundance of Stx phages in the extraintestinal environment confirm the role of Stx phages as a reservoir of stx in the environment. PMID:20622134

  18. Validation of a DNA IQ-based extraction method for TECAN robotic liquid handling workstations for processing casework.

    PubMed

    Frégeau, Chantal J; Lett, C Marc; Fourney, Ron M

    2010-10-01

    A semi-automated DNA extraction process for casework samples based on the Promega DNA IQ™ system was optimized and validated on TECAN Genesis 150/8 and Freedom EVO robotic liquid handling stations configured with fixed tips and a TECAN TE-Shake™ unit. The use of an orbital shaker during the extraction process promoted efficiency with respect to DNA capture, magnetic bead/DNA complex washes and DNA elution. Validation studies determined the reliability and limitations of this shaker-based process. Reproducibility with regards to DNA yields for the tested robotic workstations proved to be excellent and not significantly different than that offered by the manual phenol/chloroform extraction. DNA extraction of animal:human blood mixtures contaminated with soil demonstrated that a human profile was detectable even in the presence of abundant animal blood. For exhibits containing small amounts of biological material, concordance studies confirmed that DNA yields for this shaker-based extraction process are equivalent or greater to those observed with phenol/chloroform extraction as well as our original validated automated magnetic bead percolation-based extraction process. Our data further supports the increasing use of robotics for the processing of casework samples. Crown Copyright © 2009. Published by Elsevier Ireland Ltd. All rights reserved.

  19. Lidar Based Emissions Measurement at the Whole Facility Scale: Method and Error Analysis

    USDA-ARS?s Scientific Manuscript database

    Particulate emissions from agricultural sources vary from dust created by operations and animal movement to the fine secondary particulates generated from ammonia and other emitted gases. The development of reliable facility emission data using point sampling methods designed to characterize regiona...

  20. Radioactivity contents in dicalcium phosphate and the potential radiological risk to human populations.

    PubMed

    Casacuberta, N; Masqué, P; Garcia-Orellana, J; Bruach, J M; Anguita, M; Gasa, J; Villa, M; Hurtado, S; Garcia-Tenorio, R

    2009-10-30

    Potentially harmful phosphate-based products derived from the wet acid digestion of phosphate rock represent one of the most serious problems facing the phosphate industry. This is particularly true for dicalcium phosphate (DCP), a food additive produced from either sulphuric acid or hydrochloric acid digestion of raw rock material. This study determined the natural occurring radionuclide concentrations of 12 DCP samples and 4 tricalcium phosphate (TCP) samples used for animal and human consumption, respectively. Metal concentrations (Al, Fe, Zn, Cd, Cr, As, Hg, Pb and Mg) were also determined. Samples were grouped into three different clusters (A, B, C) based on their radionuclide content. Whereas group A is characterized by high activities of 238U, 234U (approximately 10(3) Bq kg(-1)), 210Pb (2 x 10(3) Bq kg(-1)) and (210)Po ( approximately 800 Bq kg(-1)); group B presents high activities of (238)U, (234)U and (230)Th (approximately 10(3) Bq kg(-1)). Group C was characterized by very low activities of all radionuclides (< 50 Bq kg(-1)). Differences between the two groups of DCP samples for animal consumption (groups A and B) were related to the wet acid digestion method used, with group A samples produced from hydrochloric acid digestion, and group B samples produced using sulphuric acid. Group C includes more purified samples required for human consumption. High radionuclide concentrations in some DCP samples (reaching 2 x 10(3) and 10(3) Bq kg(-1) of 210Pb and 210Po, respectively) may be of concern due to direct or indirect radiological exposure via ingestion. Our experimental results based on 210Pb and 210Po within poultry consumed by humans, suggest that the maximum radiological doses are 11 +/- 2 microSv y(-1). While these results suggest that human health risks are small, additional testing should be conducted.

  1. Pre-analytic evaluation of volumetric absorptive microsampling and integration in a mass spectrometry-based metabolomics workflow.

    PubMed

    Volani, Chiara; Caprioli, Giulia; Calderisi, Giovanni; Sigurdsson, Baldur B; Rainer, Johannes; Gentilini, Ivo; Hicks, Andrew A; Pramstaller, Peter P; Weiss, Guenter; Smarason, Sigurdur V; Paglia, Giuseppe

    2017-10-01

    Volumetric absorptive microsampling (VAMS) is a novel approach that allows single-drop (10 μL) blood collection. Integration of VAMS with mass spectrometry (MS)-based untargeted metabolomics is an attractive solution for both human and animal studies. However, to boost the use of VAMS in metabolomics, key pre-analytical questions need to be addressed. Therefore, in this work, we integrated VAMS in a MS-based untargeted metabolomics workflow and investigated pre-analytical strategies such as sample extraction procedures and metabolome stability at different storage conditions. We first evaluated the best extraction procedure for the polar metabolome and found that the highest number and amount of metabolites were recovered upon extraction with acetonitrile/water (70:30). In contrast, basic conditions (pH 9) resulted in divergent metabolite profiles mainly resulting from the extraction of intracellular metabolites originating from red blood cells. In addition, the prolonged storage of blood samples at room temperature caused significant changes in metabolome composition, but once the VAMS devices were stored at - 80 °C, the metabolome remained stable for up to 6 months. The time used for drying the sample did also affect the metabolome. In fact, some metabolites were rapidly degraded or accumulated in the sample during the first 48 h at room temperature, indicating that a longer drying step will significantly change the concentration in the sample. Graphical abstract Volumetric absorptive microsampling (VAMS) is a novel technology that allows single-drop blood collection and, in combination with mass spectrometry (MS)-based untargeted metabolomics, represents an attractive solution for both human and animal studies. In this work, we integrated VAMS in a MS-based untargeted metabolomics workflow and investigated pre-analytical strategies such as sample extraction procedures and metabolome stability at different storage conditions. The latter revealed that prolonged storage of blood samples at room temperature caused significant changes in metabolome composition, but if VAMS devices were stored at - 80 °C, the metabolome remained stable for up to 6 months.

  2. The Evolution of Tyrosine-Recombinase Elements in Nematoda

    PubMed Central

    Szitenberg, Amir; Koutsovoulos, Georgios; Blaxter, Mark L.; Lunt, David H.

    2014-01-01

    Transposable elements can be categorised into DNA and RNA elements based on their mechanism of transposition. Tyrosine recombinase elements (YREs) are relatively rare and poorly understood, despite sharing characteristics with both DNA and RNA elements. Previously, the Nematoda have been reported to have a substantially different diversity of YREs compared to other animal phyla: the Dirs1-like YRE retrotransposon was encountered in most animal phyla but not in Nematoda, and a unique Pat1-like YRE retrotransposon has only been recorded from Nematoda. We explored the diversity of YREs in Nematoda by sampling broadly across the phylum and including 34 genomes representing the three classes within Nematoda. We developed a method to isolate and classify YREs based on both feature organization and phylogenetic relationships in an open and reproducible workflow. We also ensured that our phylogenetic approach to YRE classification identified truncated and degenerate elements, informatively increasing the number of elements sampled. We identified Dirs1-like elements (thought to be absent from Nematoda) in the nematode classes Enoplia and Dorylaimia indicating that nematode model species do not adequately represent the diversity of transposable elements in the phylum. Nematode Pat1-like elements were found to be a derived form of another Pat1-like element that is present more widely in animals. Several sequence features used widely for the classification of YREs were found to be homoplasious, highlighting the need for a phylogenetically-based classification scheme. Nematode model species do not represent the diversity of transposable elements in the phylum. PMID:25197791

  3. The evolution of tyrosine-recombinase elements in Nematoda.

    PubMed

    Szitenberg, Amir; Koutsovoulos, Georgios; Blaxter, Mark L; Lunt, David H

    2014-01-01

    Transposable elements can be categorised into DNA and RNA elements based on their mechanism of transposition. Tyrosine recombinase elements (YREs) are relatively rare and poorly understood, despite sharing characteristics with both DNA and RNA elements. Previously, the Nematoda have been reported to have a substantially different diversity of YREs compared to other animal phyla: the Dirs1-like YRE retrotransposon was encountered in most animal phyla but not in Nematoda, and a unique Pat1-like YRE retrotransposon has only been recorded from Nematoda. We explored the diversity of YREs in Nematoda by sampling broadly across the phylum and including 34 genomes representing the three classes within Nematoda. We developed a method to isolate and classify YREs based on both feature organization and phylogenetic relationships in an open and reproducible workflow. We also ensured that our phylogenetic approach to YRE classification identified truncated and degenerate elements, informatively increasing the number of elements sampled. We identified Dirs1-like elements (thought to be absent from Nematoda) in the nematode classes Enoplia and Dorylaimia indicating that nematode model species do not adequately represent the diversity of transposable elements in the phylum. Nematode Pat1-like elements were found to be a derived form of another Pat1-like element that is present more widely in animals. Several sequence features used widely for the classification of YREs were found to be homoplasious, highlighting the need for a phylogenetically-based classification scheme. Nematode model species do not represent the diversity of transposable elements in the phylum.

  4. Validation of a PCR-based method for the detection of various rendered materials in feedstuffs using a forensic DNA extraction kit.

    PubMed

    Myers, Michael J; Yancy, Haile F; Araneta, Michael; Armour, Jennifer; Derr, Janice; Hoostelaere, Lawrence A D; Farmer, Doris; Jackson, Falana; Kiessling, William M; Koch, Henry; Lin, Huahua; Liu, Yan; Mowlds, Gabrielle; Pinero, David; Riter, Ken L; Sedwick, John; Shen, Yuelian; Wetherington, June; Younkins, Ronsha

    2006-01-01

    A method trial was initiated to validate the use of a commercial DNA forensic kit to extract DNA from animal feed as part of a PCR-based method. Four different PCR primer pairs (one bovine pair, one porcine pair, one ovine primer pair, and one multispecies pair) were also evaluated. Each laboratory was required to analyze a total of 120 dairy feed samples either not fortified (control, true negative) or fortified with bovine meat and bone meal, porcine meat and bone meal (PMBM), or lamb meal. Feeds were fortified with the animal meals at a concentration of 0.1% (wt/wt). Ten laboratories participated in this trial, and each laboratory was required to evaluate two different primer pairs, i.e., each PCR primer pair was evaluated by five different laboratories. The method was considered to be validated for a given animal source when three or more laboratories achieved at least 97% accuracy (29 correct of 30 samples for 96.7% accuracy, rounded up to 97%) in detecting the fortified samples for that source. Using this criterion, the method was validated for the bovine primer because three laboratories met the criterion, with an average accuracy of 98.9%. The average false-positive rate was 3.0% in these laboratories. A fourth laboratory was 80% accurate in identifying the samples fortified with bovine meat and bone meal. A fifth laboratory was not able to consistently extract the DNA from the feed samples and did not achieve the criterion for accuracy for either the bovine or multispecies PCR primers. For the porcine primers, the method was validated, with four laboratories meeting the criterion for accuracy with an average accuracy of 99.2%. The fifth laboratory had a 93.3% accuracy outcome for the porcine primer. Collectively, these five laboratories had a 1.3% false-positive rate for the porcine primer. No laboratory was able to meet the criterion for accuracy with the ovine primers, most likely because of problems with the synthesis of the primer pair; none of the positive control DNA samples could be detected with the ovine primers. The multispecies primer pair was validated in three laboratories for use with bovine meat and bone meal and lamb meal but not with PMBM. The three laboratories had an average accuracy of 98.9% for bovine meat and bone meal, 97.8% for lamb meal, and 63.3% for PMBM. When examined on an individual laboratory basis, one of these four laboratories could not identify a single feed sample containing PMBM by using the multispecies primer, whereas the other laboratory identified only one PMBM-fortified sample, suggesting that the limit of detection for PMBM with this primer pair is around 0.1% (wt/wt). The results of this study demonstrated that the DNA forensic kit can be used to extract DNA from animal feed, which can then be used for PCR analysis to detect animal-derived protein present in the feed sample.

  5. Evaluating pasture and soil allowance of manganese for Kajli rams grazing in semi-arid environment.

    PubMed

    Khan, Zafar Iqbal; Ahmad, Kafeel; Ashraf, Muhammad; Naqvi, Syed Ali Hassan; Seidavi, Alireza; Akram, Nudrat Aisha; Laudadio, Vito; Tufarelli, Vincenzo

    2015-03-01

    The current research on the manganese (Mn) transfer from soil to plant as well as to grazing Kajli rams in the form of sampling periods was carried out under semi-arid environmental conditions. Forage, soil and blood plasma samples were collected during 4 months of the year after a 1-month interval, and Mn concentrations were assessed after wet digestion using an atomic absorption spectrophotometer. Results showed that Mn concentration in soil ranged from 48.28 to 59.44 mg/kg, with incoherent augment and decline across sampling periods, and effect of sampling period on soil Mn was also found to be significant (P < 0.05). The mean levels of Mn in soil appeared higher than the critical value and sufficient for forage crop requirement. The Mn concentration in forage ranged between 24.8 and 37.2 mg/kg, resulting deficient based on the requirement allowance of Mn for livestock grazing animals, therein with almost unchanged forage Mn concentration. The Mn values in blood plasma of rams varied from 0.066 to 0.089 mg/l, with a consistent increase based on sampling period, and the effect of sampling periods on plasma Mn was found to be highly significant (P < 0.05). The Mn levels in ram blood plasma were lesser than the normal level suggesting reasonable need for supplementation. Our study revealed the role of Mn availability in soil and plant species amassing capability on the transport of Mn in the soil-plant-animal system. Results indicated a much higher accumulation rate at the sampling characterized by vegetation dominated by legumes in comparison to grasses, crop residues and mixed pasture and a pronounced seasonal supply of Mn at the four sampling period of grazing land of diverse botanical composition.

  6. Improving the accuracy of livestock distribution estimates through spatial interpolation.

    PubMed

    Bryssinckx, Ward; Ducheyne, Els; Muhwezi, Bernard; Godfrey, Sunday; Mintiens, Koen; Leirs, Herwig; Hendrickx, Guy

    2012-11-01

    Animal distribution maps serve many purposes such as estimating transmission risk of zoonotic pathogens to both animals and humans. The reliability and usability of such maps is highly dependent on the quality of the input data. However, decisions on how to perform livestock surveys are often based on previous work without considering possible consequences. A better understanding of the impact of using different sample designs and processing steps on the accuracy of livestock distribution estimates was acquired through iterative experiments using detailed survey. The importance of sample size, sample design and aggregation is demonstrated and spatial interpolation is presented as a potential way to improve cattle number estimates. As expected, results show that an increasing sample size increased the precision of cattle number estimates but these improvements were mainly seen when the initial sample size was relatively low (e.g. a median relative error decrease of 0.04% per sampled parish for sample sizes below 500 parishes). For higher sample sizes, the added value of further increasing the number of samples declined rapidly (e.g. a median relative error decrease of 0.01% per sampled parish for sample sizes above 500 parishes. When a two-stage stratified sample design was applied to yield more evenly distributed samples, accuracy levels were higher for low sample densities and stabilised at lower sample sizes compared to one-stage stratified sampling. Aggregating the resulting cattle number estimates yielded significantly more accurate results because of averaging under- and over-estimates (e.g. when aggregating cattle number estimates from subcounty to district level, P <0.009 based on a sample of 2,077 parishes using one-stage stratified samples). During aggregation, area-weighted mean values were assigned to higher administrative unit levels. However, when this step is preceded by a spatial interpolation to fill in missing values in non-sampled areas, accuracy is improved remarkably. This counts especially for low sample sizes and spatially even distributed samples (e.g. P <0.001 for a sample of 170 parishes using one-stage stratified sampling and aggregation on district level). Whether the same observations apply on a lower spatial scale should be further investigated.

  7. Self-clearance from BVDV infections--a frequent finding in dairy herds in an endemically infected region in Peru.

    PubMed

    Ståhl, K; Lindberg, A; Rivera, H; Ortiz, C; Moreno-López, J

    2008-03-17

    In this cross-sectional study, a stratified two-stage random sampling procedure was employed to select 221 dairy herds for bulk tank milk (BTM) sampling, and a subset of 55 dairy herds for individual blood sampling of a number of young animals (spot test), to predict presence or absence of current BVDV infection, and for data collection. The prediction was based on the high probability of seropositivity in groups of animals where PI animals are present because of the efficient spread of virus from PI animals to the surrounding group. BTM samples were collected in August 2003 (n=192) and February 2004 (n=195), and the 55 herds selected for spot testing and data collection were visited in December 2003. All samples were tested for presence of BVDV specific antibodies using a commercial indirect ELISA (SVANOVA Biotech AB, Uppsala, Sweden). The results demonstrated a very high level of exposure to BVDV in the region, and the proportion of herds with high antibody levels in the BTM was above 95% on both occasions. Despite this, almost two thirds of the herds had spot test results indicating absence of current infection, suggesting a high probability of self-clearance. A logistic regression model with the results from the spot tests as dependent variable was used to investigate possible herd and management factors associated with self-clearance, and suggested that this may occur regardless of herd size. Even though it is well established that the process of identification and elimination of PI animals is required within a systematic BVDV eradication programme, the present study strongly suggests that many herds may be cleared without intervention even in regions with high cattle density and high BVDV prevalence. Consequently, in any BVDV infected population (regardless of the herd-level BVDV seroprevalence), and at any given point of time, a large proportion of the herds will be free from infection due to self-clearance. Self-clearance is therefore a process that works in favour of any effort to control BVDV, which should be taken into account when planning and assessing the cost-effectiveness of a systematic control programme.

  8. Diversity of Leptospira spp. in Rats and Environment from Urban Areas of Sarawak, Malaysia

    PubMed Central

    Pui, Chai Fung; Apun, Kasing; Su'ut, Lela

    2017-01-01

    Various prevalence studies on Leptospira in animals and humans, as well as environmental samples, had been conducted worldwide, including Malaysia. However, limited studies have been documented on the presence of pathogenic, intermediate, and saprophytic Leptospira in selected animals and environments. This study was therefore conducted to detect Leptospira spp. in rats, soil, and water from urban areas of Sarawak using the polymerase chain reaction (PCR) method. A total of 107 rats, 292 soil samples, and 324 water samples were collected from April 2014 to February 2015. Pathogenic Leptospira was present in 5.6% (6/107) of rats, 11.6% (34/292) of soil samples, and 1.9% (6/324) of water samples. Intermediate Leptospira was present in 2.7% (8/292) of soil samples and 1.9% (6/324) of water samples. Saprophytic Leptospira was present in 10.3% (11/107) of rats, 1.4% (4/292) of soil samples, and 0.3% (1/324) of water samples. From this study, 76 Leptospira spp. were isolated. Based on DNA sequencing, the dominant Leptospira spp. circulating in urban areas of Sarawak are pathogenic Leptospira noguchii, intermediate Leptospira wolffii serovar Khorat, and saprophytic Leptospira meyeri, respectively. Overall, this study provided important surveillance data on the prevalence of Leptospira spp. from rats and the environment, with dominant local serovars in urban areas of Sarawak. PMID:28348601

  9. Diversity of Leptospira spp. in Rats and Environment from Urban Areas of Sarawak, Malaysia.

    PubMed

    Pui, Chai Fung; Bilung, Lesley Maurice; Apun, Kasing; Su'ut, Lela

    2017-01-01

    Various prevalence studies on Leptospira in animals and humans, as well as environmental samples, had been conducted worldwide, including Malaysia. However, limited studies have been documented on the presence of pathogenic, intermediate, and saprophytic Leptospira in selected animals and environments. This study was therefore conducted to detect Leptospira spp. in rats, soil, and water from urban areas of Sarawak using the polymerase chain reaction (PCR) method. A total of 107 rats, 292 soil samples, and 324 water samples were collected from April 2014 to February 2015. Pathogenic Leptospira was present in 5.6% (6/107) of rats, 11.6% (34/292) of soil samples, and 1.9% (6/324) of water samples. Intermediate Leptospira was present in 2.7% (8/292) of soil samples and 1.9% (6/324) of water samples. Saprophytic Leptospira was present in 10.3% (11/107) of rats, 1.4% (4/292) of soil samples, and 0.3% (1/324) of water samples. From this study, 76 Leptospira spp. were isolated. Based on DNA sequencing, the dominant Leptospira spp. circulating in urban areas of Sarawak are pathogenic Leptospira noguchii , intermediate Leptospira wolffii serovar Khorat, and saprophytic Leptospira meyeri , respectively. Overall, this study provided important surveillance data on the prevalence of Leptospira spp. from rats and the environment, with dominant local serovars in urban areas of Sarawak.

  10. The first study of molecular prevalence and species characterization of Cryptosporidium in free-range chicken (Gallus gallus domesticus) from Brazil.

    PubMed

    Ewald, Maria Paula de Carvalho; Martins, Felippe Danyel Cardoso; Caldart, Eloiza Teles; Vieira, Fernando Emmanuel Gonçalves; Yamamura, Milton Hissashi; Sasse, João Pedro; Barros, Luiz Daniel de; Freire, Roberta Lemos; Navarro, Italmar Teodorico; Garcia, João Luis

    2017-01-01

    Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

  11. Estimating occupancy and abundance of stream amphibians using environmental DNA from filtered water samples

    USGS Publications Warehouse

    Pilliod, David S.; Goldberg, Caren S.; Arkle, Robert S.; Waits, Lisette P.

    2013-01-01

    Environmental DNA (eDNA) methods for detecting aquatic species are advancing rapidly, but with little evaluation of field protocols or precision of resulting estimates. We compared sampling results from traditional field methods with eDNA methods for two amphibians in 13 streams in central Idaho, USA. We also evaluated three water collection protocols and the influence of sampling location, time of day, and distance from animals on eDNA concentration in the water. We found no difference in detection or amount of eDNA among water collection protocols. eDNA methods had slightly higher detection rates than traditional field methods, particularly when species occurred at low densities. eDNA concentration was positively related to field-measured density, biomass, and proportion of transects occupied. Precision of eDNA-based abundance estimates increased with the amount of eDNA in the water and the number of replicate subsamples collected. eDNA concentration did not vary significantly with sample location in the stream, time of day, or distance downstream from animals. Our results further advance the implementation of eDNA methods for monitoring aquatic vertebrates in stream habitats.

  12. Tick- and fly-borne bacteria in ungulates: the prevalence of Anaplasma phagocytophilum, haemoplasmas and rickettsiae in water buffalo and deer species in Central Europe, Hungary.

    PubMed

    Hornok, Sándor; Sugár, László; Fernández de Mera, Isabel G; de la Fuente, José; Horváth, Gábor; Kovács, Tibor; Micsutka, Attila; Gönczi, Enikő; Flaisz, Barbara; Takács, Nóra; Farkas, Róbert; Meli, Marina L; Hofmann-Lehmann, Regina

    2018-03-20

    Hunting constitutes an important industry in Europe. However, data on the prevalence of vector-borne bacteria in large game animal species are lacking from several countries. Blood or spleen samples (239 and 270, respectively) were taken from red, fallow and roe deer, as well as from water buffaloes, mouflons and wild boars in Hungary, followed by DNA extraction and molecular analyses for Anaplasma phagocytophilum, haemoplasmas and rickettsiae. Based on blood samples, the prevalence rate of A. phagocytophilum infection was significantly higher in red deer (97.9%) than in fallow deer (72.7%) and roe deer (60%), and in all these compared to mouflons (6.3%). In addition, 39.2% of the spleen samples from wild boars were PCR positive for A. phagocytophilum, but none of the buffalos. Based on blood samples, the prevalence rates of both Mycoplasma wenyonii (Mw) and 'Candidatus M. haemobos' (CMh) infections were significantly higher in buffaloes (Mw: 91.2%; CMh: 73.3%) than in red deer (Mw: 64.6%; CMh: 45.8%), and in both of them compared to fallow deer (Mw: 30.3%; CMh: 9.1%) and roe deer (Mw: 20%; CMh: 1.5%). The prevalence of Mw and CMh infection significantly correlated with the body sizes of these hosts. Furthermore, Mw was significantly more prevalent than CMh in buffaloes, red and roe deer. Mycoplasma ovis was detected in mouflons, M. suis in wild boars, R. helvetica in one fallow deer and one mouflon, and an unidentified Rickettsia sp. in a fallow deer. Forest-dwelling game animal species were found to be important carriers of A. phagocytophilum. In contrast, animals grazing grassland (i.e. buffaloes) were less likely to get infected with this Ixodes ricinus-borne pathogen. Water buffaloes, deer species, mouflons and wild boars harbored haemoplasmas that may affect domestic ungulates. Evaluated animals with larger body size had significantly higher prevalence of infection with haemoplasmas compared to smaller deer species. The above host species rarely carried rickettsiae.

  13. Herd-level prevalence of Map infection in dairy herds of southern Chile determined by culture of environmental fecal samples and bulk-tank milk qPCR.

    PubMed

    Kruze, J; Monti, G; Schulze, F; Mella, A; Leiva, S

    2013-09-01

    Paratuberculosis, an infectious disease of domestic and wild ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map), is an economically important disease in dairy herds worldwide. In Chile the disease has been reported in domestic and wildlife animals. However, accurate and updated estimations of the herd-prevalence in cattle at national or regional level are not available. The objectives of this study were to determine the herd-level prevalence of dairy herds with Map infected animals of Southern Chile, based on two diagnostic tests: culture of environmental fecal samples and bulk-tank milk qPCR. Two composite environmental fecal samples and one bulk-tank milk sample were collected during September 2010 and September 2011 from 150 dairy farms in Southern Chile. Isolation of Map from environmental fecal samples was done by culture of decontaminated samples on a commercial Herrold's Egg Yolk Medium (HEYM) with and without mycobactin J. Suspicious colonies were confirmed to be Map by conventional IS900 PCR. Map detection in bulk-tank milk samples was done by real time IS900 PCR assay. PCR-confirmed Map was isolated from 58 (19.3%) of 300 environmental fecal samples. Holding pens and manure storage lagoons were the two more frequent sites found positive for Map, representing 35% and 33% of total positive samples, respectively. However, parlor exits and cow alleyways were the two sites with the highest proportion of positive samples (40% and 32%, respectively). Herd prevalence based on environmental fecal culture was 27% (true prevalence 44%) compared to 49% (true prevalence 87%) based on bulk-tank milk real time IS900 PC. In both cases herd prevalence was higher in large herds (>200 cows). These results confirm that Map infection is wide spread in dairy herds in Southern Chile with a rough herd-level prevalence of 28-100% depending on the herd size, and that IS900 PCR on bulk-tank milk samples is more sensitive than environmental fecal culture to detect Map-infected dairy herds. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Titration Techniques

    NASA Astrophysics Data System (ADS)

    Jacobsen, Jerrold J.; Houston Jetzer, Kelly; Patani, Néha; Zimmerman, John; Zweerink, Gerald

    1995-07-01

    Significant attention is paid to the proper technique for reading a meniscus. Video shows meniscus-viewing techniques for colorless and dark liquids and the consequences of not reading a meniscus at eye level. Lessons are provided on approaching the end point, focusing on end point colors produced via different commonly used indicators. The concept of a titration curve is illustrated by means of a pH meter. Carefully recorded images of the entire range of meniscus values in a buret, pipet, and graduated cylinder are included so that you can show your students, in lecture or pre-lab discussion, any meniscus and discuss how to read the buret properly. These buret meniscus values are very carefully recorded at the rate of one video frame per hundredth of a milliliter, so that an image showing any given meniscus value can be obtained. These images can be easily incorporated into a computer-based multimedia environment for testing or meniscus-reading exercises. Two of the authors have used this technique and found the exercise to be very well received by their students. Video on side two shows nearly 100 "bloopers", demonstrating both the right way and wrong ways to do tasks associated with titration. This material can be used in a variety of situations: to show students the correct way to do something; to test students by asking them "What is this person doing wrong?"; or to develop multimedia, computer-based lessons. The contents of Titration Techniques are listed below: Side 1 Titration: what it is. A simple titration; Acid-base titration animation; A brief redox titration; Redox titration animation; A complete acid-base titration. Titration techniques. Hand technique variations; Stopcock; Using a buret to measure liquid volumes; Wait before reading meniscus; Dirty and clean burets; Read meniscus at eye level (see Fig. 1); Meniscus viewing techniques--light colored liquids; Meniscus viewing techniques--dark liquids; Using a magnetic stirrer; Rough titration; Significant figures; Approaching the end point; End point colors; Titration with a pH meter; Titration curves; Colors of indicators. Meniscus values. Buret meniscus values; Pipet meniscus values; Graduated cylinder meniscus values. Side 2"Bloopers". Introducing the people; Titration animation; Inspecting the buret; Rinsing the buret with water; Preparing a solid sample; Obtaining a liquid sample; Delivering a liquid sample with a Mohr pipet; Pipetting a liquid sample with a Mohr pipet; Rinsing the Mohr pipet with sample; Using the Mohr pipet to transfer sample; Delivering a liquid sample with a volumetric pipet; Pipetting a liquid sample with a volumetric pipet; Rinsing the volumetric pipet with sample; Using the volumetric pipet to transfer sample; Obtaining the titrant; Rinsing the buret with titrant; Filling the buret with titrant; Adding the indicator; The initial reading; Beginning the titration; Delivering titrant; The final reading. Figure 3. Near the end point a single drop of titrant can cause a lasting color change.

  15. AFNOR validation of Premi Test, a microbiological-based screening tube-test for the detection of antimicrobial residues in animal muscle tissue.

    PubMed

    Gaudin, Valerie; Juhel-Gaugain, Murielle; Morétain, Jean-Pierre; Sanders, Pascal

    2008-12-01

    Premi Test contains viable spores of a strain of Bacillus stearothermophilus which is sensitive to antimicrobial residues, such as beta-lactams, tetracyclines, macrolides and sulphonamides. The growth of the strain is inhibited by the presence of antimicrobial residues in muscle tissue samples. Premi Test was validated according to AFNOR rules (French Association for Normalisation). The AFNOR validation was based on the comparison of reference methods (French Official method, i.e. four plate test (FPT) and the STAR protocol (five plate test)) with the alternative method (Premi Test). A preliminary study was conducted in an expert laboratory (Community Reference Laboratory, CRL) on both spiked and incurred samples (field samples). Several method performance criteria (sensitivity, specificity, relative accuracy) were estimated and are discussed, in addition to detection capabilities. Adequate agreement was found between the alternative method and the reference methods. However, Premi Test was more sensitive to beta-lactams and sulphonamides than the FPT. Subsequently, a collaborative study with 11 laboratories was organised by the CRL. Blank and spiked meat juice samples were sent to participants. The expert laboratory (CRL) statistically analysed the results. It was concluded that Premi Test could be used for the routine determination of antimicrobial residues in muscle of different animal origin with acceptable analytical performance. The detection capabilities of Premi Test for beta-lactams (amoxicillin, ceftiofur), one macrolide (tylosin) and tetracycline were at the level of the respective maximum residue limits (MRL) in muscle samples or even lower.

  16. Multidrug-resistant Salmonella Typhimurium in Four Animal Facilities

    PubMed Central

    Wright, Jennifer G.; Tengelsen, Leslie A.; Smith, Kirk E.; Bender, Jeff B.; Frank, Rodney K.; Grendon, John H.; Rice, Daniel H.; Thiessen, Ann Marie B.; Gilbertson, Catherine Jo; Sivapalasingam, Sumathi; Barrett, Timothy J.; Besser, Thomas E.; Hancock, Dale D.

    2005-01-01

    In 1999 and 2000, 3 state health departments reported 4 outbreaks of gastrointestinal illness due to Salmonella enterica serotype Typhimurium in employees, clients, and client animals from 3 companion animal veterinary clinics and 1 animal shelter. More than 45 persons and companion animals became ill. Four independent investigations resulted in the testing of 19 human samples and >200 animal samples; 18 persons and 36 animals were culture-positive for S. Typhimurium. One outbreak was due to multidrug-resistant S. Typhimurium R-type ACKSSuT, while the other 3 were due to multidrug-resistant S. Typhimurium R-type ACSSuT DT104. This report documents nosocomial transmission of S. Typhimurium and demonstrates that companion animal facilities may serve as foci of transmission for salmonellae between animals and humans if adequate precautions are not followed. PMID:16102313

  17. Placement of intracoelomic radio transmitters and silicone passive sampling devices in northern leopard frogs (Lithobates pipiens)

    USGS Publications Warehouse

    Yaw, Taylor; Swanson, Jennifer E; Pierce, Clay; Muths, Erin L.; Smalling, Kelly; Vandever, Mark; Zaffarano, Bianca Anne

    2017-01-01

    Historically, wetland toxin exposure studies have relied on single time point samples from stationary sampling devices. Development of passive sampling devices (PSDs) that can be attached to individual animals within wetland habitats has greatly improved in recent years, presenting an innovative sampling technology that can potentially yield individual-specific, quantifiable data about chemical exposure. In this study, silicone based PSDs were attached to the ventral skin of 20 northern leopard frogs (Lithobates pipiens) with polypropylene sutures after radio transmitters had been surgically implanted into the coleomic cavity. After a recovery period frogs were released back into the wetland habitat where they were acquired. The animals were located daily using radio telemetry to assess how long PSDs would remain attached in the frogs' natural habitat. After one week, PSDs remained on 18 of the original 20 frogs. At 2 weeks 17 frogs were recovered and no PSDs remained attached. Although valuable data can be obtained over a short time period, more research will be necessary to demonstrate effectiveness of externally attaching silicone PSDs to northern leopard frogs for time periods longer than 1-2 weeks.

  18. Multiresidue analysis of sulfonamides, quinolones, and tetracyclines in animal tissues by ultra-high performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Zhang, Zhiwen; Li, Xiaowei; Ding, Shuangyang; Jiang, Haiyang; Shen, Jianzhong; Xia, Xi

    2016-08-01

    A multiresidue method for the efficient identification and quantification of 38 compounds from 3 different classes of antibiotics (tetracyclines, sulfonamides, and quinolones) in animal tissues has been developed. The method optimization involved the selection of extraction solutions, comparison of different solid-phase extraction cartridges and different mobile phases. As a result, the samples were extracted with Mcllvaine and phosphate buffers, followed by clean-up step based on solid-phase extraction with Oasis HLB cartridge. All compounds were determined by ultra-high performance liquid chromatography-tandem mass spectrometry, in one single injection with a chromatographic run time of only 9min. The method efficiency was evaluated in 5 tissues including muscle, liver, and kidney, and the mean recoveries ranged from 54% to 102%, with inter-day relative standard deviation lower than 14%. The limits of quantification were between 0.5 and 10μg/kg, which were satisfactory to support future surveillance monitoring. The developed method was applied to the analysis of swine liver and chicken samples from local markets, and sulfamethazine was the most commonly detected compound in the animal samples, with the highest residue level of 998μg/kg. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Development of a novel lateral flow assay for detection of African swine fever in blood.

    PubMed

    Sastre, P; Gallardo, C; Monedero, A; Ruiz, T; Arias, M; Sanz, A; Rueda, P

    2016-09-15

    African swine fever (ASF) is a viral infectious disease of domestic and wild suids of all breeds and ages, causing a wide range of hemorrhagic syndromes and frequently characterized by high mortality. The disease is endemic in Sub-Saharan Africa and Sardinia. Since 2007, it has also been present in different countries of Eastern Europe, where control measures have not been effective so far. The continued spread poses a serious threat to the swine industry worldwide. In the absence of vaccine, early detection of infected animals is of paramount importance for control of the outbreak, to prevent the transmission of the virus to healthy animals and subsequent spreading of the disease. Current laboratory diagnosis is mainly based on virological methods (antigen and genome detection) and serodiagnosis. In the present work, a Lateral Flow Assay (LFA) for antigen detection has been developed and evaluated. The test is based on the use of a MAb against VP72 protein of ASFV, the major viral capsid protein and highly immunogenic. First experiments using VP72 viral and recombinant protein or inactivated culture virus showed promising results with a sensitivity similar to that of a commercially available Antigen-ELISA. Moreover, these strips were tested with blood from experimentally infected pigs and field animals and the results compared with those of PCR and Antigen-ELISA. For the experimentally infected samples, there was an excellent correlation between the LFA and the ELISA, while the PCR always showed to be more sensitive (38 % positive samples by PCR versus 27 % by LFA). The LFA was demonstrated to be positive for animals with circulating virus levels exceeding 10(4) HAU. With the field samples, once again, the PCR detected more positives than either the Antigen-ELISA or LFA, although here the number of positive samples scored by the LFA exceeded the values obtained with the Antigen-ELISA, showing 60 % positivity vs 48 % for the ELISA. For the two groups of sera, the specificity was close to 100 % indicating that hardly any false positive samples were found. The newly developed LFA allows rapid and reliable detection of ASFV, at field and laboratory level, providing a new useful tool for control programs and in situations where laboratory support and skilled personnel are limited.

  20. Robustness of survival estimates for radio-marked animals

    USGS Publications Warehouse

    Bunck, C.M.; Chen, C.-L.

    1992-01-01

    Telemetry techniques are often used to study the survival of birds and mammals; particularly whcn mark-recapture approaches are unsuitable. Both parametric and nonparametric methods to estimate survival have becn developed or modified from other applications. An implicit assumption in these approaches is that the probability of re-locating an animal with a functioning transmitter is one. A Monte Carlo study was conducted to determine the bias and variance of the Kaplan-Meier estimator and an estimator based also on the assumption of constant hazard and to eva!uate the performance of the two-sample tests associated with each. Modifications of each estimator which allow a re-Iocation probability of less than one are described and evaluated. Generallv the unmodified estimators were biased but had lower variance. At low sample sizes all estimators performed poorly. Under the null hypothesis, the distribution of all test statistics reasonably approximated the null distribution when survival was low but not when it was high. The power of the two-sample tests were similar.

  1. Domestic pigs as potential reservoirs of human and animal trypanosomiasis in Northern Tanzania

    PubMed Central

    2013-01-01

    Background Pig keeping is becoming increasingly common across sub-Saharan Africa. Domestic pigs from the Arusha region of northern Tanzania were screened for trypanosomes using PCR-based methods to examine the role of pigs as a reservoir of human and animal trypanosomiasis. Methods A total of 168 blood samples were obtained from domestic pigs opportunistically sampled across four districts in Tanzania (Babati, Mbulu, Arumeru and Dodoma) during December 2004. A suite of PCR-based methods was used to identify the species and sub-species of trypanosomes including: Internally Transcribed Sequence to identify multiple species; species specific PCR to identify T. brucei s. l. and T. godfreyi and a multiplex PCR reaction to distinguish T. b. rhodesiense from T. brucei s. l. Results Of the 168 domestic pigs screened for animal and human infective trypanosome DNA, 28 (16.7%) were infected with one or more species of trypanosome; these included: six pigs infected with Trypanosoma vivax (3.6%); three with Trypanosoma simiae (1.8%); two with Trypanosoma congolense (Forest) (1%) and four with Trypanosoma godfreyi (2.4%). Nineteen pigs were infected with Trypanosoma brucei s. l. (10.1%) of which eight were identified as carrying the human infective sub-species Trypanosoma brucei rhodesiense (4.8%). Conclusion These results show that in Tanzania domestic pigs may act as a significant reservoir for animal trypanosomiasis including the cattle pathogens T. vivax and T. congolense, the pig pathogen T. simiae, and provide a significant reservoir for T. b. rhodesiense, the causative agent of acute Rhodesian sleeping sickness. PMID:24499540

  2. Identification and molecular survey of Borrelia burgdorferi sensu lato in sika deer (Cervus nippon) from Jilin Province, north-eastern China.

    PubMed

    Zhai, Bintao; Niu, Qingli; Yang, Jifei; Liu, Zhijie; Liu, Junlong; Yin, Hong; Zeng, Qiaoying

    2017-02-01

    Lyme disease caused by Borrelia burgdorferi sensu lato (s.l.) is a common disease of domestic animals and wildlife worldwide. Sika deer is first-grade state-protected wildlife animals in China and have economic consequences for humans. It is reported that sika deer may serve as an important reservoir host for several species of B. burgdorferi s.l. and may transmit these species to humans and animals. However, little is known about the presence of Borrelia pathogens in sika deer in China. In this study, the existence and prevalence of Borrelia sp. in sika deer from four regions of Jilin Province in China was assessed. Seventy-one blood samples of sika deer were collected and tested by nested-PCRs based on 16S ribosomal RNA (16S rRNA), outer surface protein A (OspA), flagenllin (fla), and 5S-23S rRNA intergenic spacer (5S-23S rRNA) genes of B. burgdorferi s.l. Six (8.45%) samples were positive for Borrelia sp. based on sequences of 4 genes. The positive samples were detected 18 for 16S rRNA, 10 for OspA, 16 for fla and 6 for 5S-23S, with the positive rates 25.35% (95% CI=3.8-35.6), 14.08% (95% CI=3.0-21.6), 22.54% (95% CI=4.3-36.9) and 8.45% (95% CI=1.7-22.9), respectively. Sequence analysis of the positive PCR products revealed that the partial 4 genes sequences in this study were all most similar to the sequences of B. garinii and B. burgdorferi sensu stricto (s.s.), no other Borrelia genospecies were found. This is the first report of Borrelia pathogens in sika deer in China. The findings in this study indicated that sika deer as potential natural host and may spread Lyme disease pathogen to animals, ticks, and even humans. Copyright © 2016. Published by Elsevier B.V.

  3. Molecular Epidemiological Survey and Genetic Characterization of Anaplasma Species in Mongolian Livestock.

    PubMed

    Ochirkhuu, Nyamsuren; Konnai, Satoru; Odbileg, Raadan; Murata, Shiro; Ohashi, Kazuhiko

    2017-08-01

    Anaplasma species are obligate intracellular rickettsial pathogens that cause great economic loss to the animal industry. Few studies on Anaplasma infections in Mongolian livestock have been conducted. This study examined the prevalence of Anaplasma marginale, Anaplasma ovis, Anaplasma phagocytophilum, and Anaplasma bovis by polymerase chain reaction assay in 928 blood samples collected from native cattle and dairy cattle (Bos taurus), yaks (Bos grunniens), sheep (Ovis aries), and goats (Capra aegagrus hircus) in four provinces of Ulaanbaatar city in Mongolia. We genetically characterized positive samples through sequencing analysis based on the heat-shock protein groEL, major surface protein 4 (msp4), and 16S rRNA genes. Only A. ovis was detected in Mongolian livestock (cattle, yaks, sheep, and goats), with 413 animals (44.5%) positive for groEL and 308 animals (33.2%) positive for msp4 genes. In the phylogenetic tree, we separated A. ovis sequences into two distinct clusters based on the groEL gene. One cluster comprised sequences derived mainly from sheep and goats, which was similar to that in A. ovis isolates from other countries. The other divergent cluster comprised sequences derived from cattle and yaks and appeared to be newly branched from that in previously published single isolates in Mongolian cattle. In addition, the msp4 gene of A. ovis using same and different samples with groEL gene of the pathogen demonstrated that all sequences derived from all animal species, except for three sequences derived from cattle and yak, were clustered together, and were identical or similar to those in isolates from other countries. We used 16S rRNA gene sequences to investigate the genetically divergent A. ovis and identified high homology of 99.3-100%. However, the sequences derived from cattle did not match those derived from sheep and goats. The results of this study on the prevalence and molecular characterization of A. ovis in Mongolian livestock can facilitate the control of infectious diseases in livestock.

  4. [Surveillance on severe acute respiratory syndrome associated coronavirus in animals at a live animal market of Guangzhou in 2004].

    PubMed

    Wang, Ming; Jing, Huai-qi; Xu, Hui-fang; Jiang, Xiu-gao; Kan, Biao; Liu, Qi-yong; Wan, Kang-lin; Cui, Bu-yun; Zheng, Han; Cui, Zhi-gang; Yan, Mei-ying; Liang, Wei-li; Wang, Hong-xia; Qi, Xiao-bao; Li, Zhen-jun; Li, Ma-chao; Chen, Kai; Zhang, En-min; Zhang, Shou-yin; Hai, Rong; Yu, Dong-zheng; Xu, Jian-guo

    2005-02-01

    To study the prevalence of severe acute respiratory syndrome coronavirus (SARS-CoV) like virus in animals at a live animal market of Guanzhou in 2004 before and after culling of wild animal action taken by the local authority, in order to predict the re-emerging of SARS from animal originals in this region. Animals at live animal market were sampled for rectal and throat swabs in triplicate. A single step realtime reverse transcription-polymerase chain reaction (RT-PCR) diagnostic kit was performed for screening SARS-CoV like virus, the manual nested RT- PCR and DNA sequencing were performed for confirmation. Only specimens which tested positive for both of the N and P genes by nested RT-PCR were scored as positive. In 31 animals sampled in January 5 2004 before culling of wild animals at Guangdong Province, including 20 cats (Felis catus), 5 red fox (Vulpes vulpes) and 6 Lesser rice field rats (Rattus losea), 8 (25.8%) animals were tested positive for SARS-CoV like virus by RT-PCR methods, of which 4 cats, 3 red fox and one Lesser rice field rats were included. However, two weeks after culling of animals and disinfection of the market were implemented, in 119 animals sampled in January 20 2004, including 6 rabbits (Oryctolagus cuniculus), 13 cats, 46 red jungle fowl (Gallus gallus), 13 spotbill duck (Anas platyrhynchos), 10 greylag goose (Anser anser), 31 Chinese francolin (Franclinus pintadeanus), only rectal swab from one greylag goose was tested positive for SARS-CoV like virus. Furthermore, in 102 animals that including 14 greylag gooses, 3 cats, 5 rabbits, 9 spotbill duck (Anaspoecilorhyncha), 2 Chinese francolin (Franclinus pintadeanus), 8 common pheasant (Phasianus colchicus), 6 pigeons, 9 Chinese muntjac (Muntiacus reevesi), 19 wild boar (Sus scrofa), 16 Lesser rice field rats, 5 dogs, 1 mink (Mustela vison), 3 goats, 2 green peafowl (Pavo muticus) sampled in April, May, June, July, August and November, only rectal swab from one pig was tested positive. However, of 12 and 10 palm civets sampled in November and December including five of which had been at the live animals market for 2 days, none of them was tested positive. This findings revealed that animals being sampled in April, May, June, July, August and November of 2004, only one rectal swab from a pig was tested positive as SARS-CoV like virus, much lower than the results from the previous year, suggesting that the possibility of re-emerging of human infection from animal origins is low for the winter of 2004-2005.

  5. Reporting the implementation of the Three Rs in European primate and mouse research papers: are we making progress?

    PubMed

    Taylor, Katy

    2010-12-01

    It is now more than 20 years since both Council of Europe Convention ETS123 and EU Directive 86/609/EEC were introduced, to promote the implementation of the Three Rs in animal experimentation and to provide guidance on animal housing and care. It might therefore be expected that reports of the implementation of the Three Rs in animal research papers would have increased during this period. In order to test this hypothesis, a literature survey of animal-based research was conducted. A randomly-selected sample from 16 high-profile medical journals, of original research papers arising from European institutions that featured experiments which involved either mice or primates, were identified for the years 1986 and 2006 (Total sample = 250 papers). Each paper was scored out of 10 for the incidence of reporting on the implementation of Three Rs-related factors corresponding to Replacement (justification of non-use of non-animal methods), Reduction (statistical analysis of the number of animals needed) and Refinement (housing aspects, i.e. increased cage size, social housing, enrichment of cage environment and food; and procedural aspects, i.e. the use of anaesthesia, analgesia, humane endpoints, and training for procedures with positive reinforcement). There was no significant increase in overall reporting score over time, for either mouse or primate research. By 2006, mouse research papers scored an average of 0 out of a possible 10, and primate research papers scored an average of 1.5. This review provides systematic evidence that animal research is still not properly reported, and supports the call within the scientific community for action to be taken by journals to update their policies. 2010 FRAME.

  6. Application and Comparative Evaluation of Fluorescent Antibody, Immunohistochemistry and Reverse Transcription Polymerase Chain Reaction Tests for the Detection of Rabies Virus Antigen or Nucleic Acid in Brain Samples of Animals Suspected of Rabies in India.

    PubMed

    Prabhu, K Nithin; Isloor, Shrikrishna; Veeresh, B Hanchinal; Rathnamma, Doddamane; Sharada, R; Das, Lekshmi J; Satyanarayana, M L; Hegde, Nagendra R; Rahman, Sira Abdul

    2018-02-28

    Accurate and early diagnosis of animal rabies is critical for undertaking public health measures. Whereas the direct fluorescent antibody (DFA) technique is the recommended test, the more convenient, direct rapid immunochemistry test (dRIT), as well as the more sensitive, reverse transcription polymerase chain reaction (RT-PCR), have recently been employed for the laboratory diagnosis of rabies. We compared the three methods on brain samples from domestic (dog, cat, cattle, buffalo, horse, pig and goat) and wild (leopard, wolf and jackal) animals from various parts of India. Of the 257 samples tested, 167 were positive by all the three tests; in addition, 35 of the 36 decomposed samples were positive by RT-PCR. This is the first study in which such large number of animal samples have been subjected to the three tests simultaneously. The results confirm 100% corroboration between DFA and dRIT, buttress the applicability of dRIT in the simple and rapid diagnosis of rabies in animals, and reaffirm the suitability of RT-PCR for samples unfit for testing either by DFA or dRIT.

  7. An eight-year survey of the intestinal parasites of carnivores, hoofed mammals, primates, ratites and reptiles in the Ljubljana zoo in Slovenia.

    PubMed

    Kvapil, Pavel; Kastelic, Marjan; Dovc, Alenka; Bartova, Eva; Cizek, Petr; Lima, Natacha; Strus, Spela

    2017-04-21

    Problems with parasitic infections and their interspecies transmissions are common in zoological gardens and could pose serious health damage to captive animals. This study presents results of eight-year monitoring of intestinal parasites in animals from Zoo Ljubljana, Slovenia. A total of 741 faecal samples from 40 animal species were collected two to four times per year and examined microscopically. Intestinal parasites were detected in 45% of samples, with detection of helminths (Cestoda, Nematoda - Ascaridida, Enoplida, Strongylida, Oxyurida, Rhabditida and Trichurida) and protists (Apicomplexa and Ciliophora) in 25% and 13% of samples, respectively; mixed infection was found in 7% of samples. The mostly infected were ungulates (61%), followed by reptiles (44%), ratites (29%), primates (22%) and carnivores (7%). During the observation period, the number of infected animal species increased from 8 to 25. This is the first long-term monitoring study of intestinal parasites in zoo animals from Slovenia. Routine monitoring of parasitic infection and regular deworming and hygienic measures are necessary to prevent gastrointestinal infections in captive animals.

  8. Prevalence of Salmonella enterica serovar Albany in captive zoo wild animals in the Culiacán Zoo in Mexico.

    PubMed

    Silva-Hidalgo, Gabriela; López-Moreno, Héctor Samuel; Ortiz-Navarrete, Vianney Francisco; Alpuche-Aranda, Celia; Rendón-Maldonado, José Guadalupe; López-Valenzuela, José Angel; López-Valenzuela, Martin; Juárez-Barranco, Felipe

    2013-03-01

    Salmonellosis is an important zoonotic disease but little is known about the role that free-living animals play as carriers of this pathogen. Moreover, the primary route of infection in the wild needs to be elucidated. The aim of this study was to determine the source and the route of transmission of Salmonella enterica serovar Albany (S. Albany) infection in captive zoo wild animals in the Culiacán Zoo. A total of 267 samples were analyzed including 220 fecal samples from zoo animals, 15 fecal samples from rodents, 5 pooled samples each of two insects (Musca domestica and Periplaneta americana), and 22 samples of animal feed. We detected S. Albany in 28 (10.5%) of the samples analyzed, including in samples from raw chicken meat. Characterization of isolates was performed by serotyping and pulsed-field gel electrophoresis. All isolates shared a single pulsed-field gel electrophoresis profile, indicating a possible common origin. These data suggest that the infected meat consumed by the wild felines was the primary source of infection in this zoo. It is likely that the pathogen was shed in the feces and disseminated by insects and rats to other locations in the zoo.

  9. Tracking animals in freshwater with electronic tags: past, present and future

    USGS Publications Warehouse

    Cooke, Steven J.; Midwood, Jonathan D.; Thiem, Jason D.; Klimley, Peter; Lucas, Martyn C.; Thorstad, Eva B.; Eiler, John; Holbrook, Chris; Ebner, Brendan C.

    2013-01-01

    Considerable technical developments over the past half century have enabled widespread application of electronic tags to the study of animals in the wild, including in freshwater environments. We review the constraints associated with freshwater telemetry and biologging and the technical developments relevant to their use. Technical constraints for tracking animals are often influenced by the characteristics of the animals being studied and the environment they inhabit. Collectively, they influence which and how technologies can be used and their relative effectiveness. Although radio telemetry has historically been the most commonly used technology in freshwater, passive integrated transponder (PIT) technology, acoustic telemetry and biologgers are becoming more popular. Most telemetry studies have focused on fish, although an increasing number have focused on other taxa, such as turtles, crustaceans and molluscs. Key technical developments for freshwater systems include: miniaturization of tags for tracking small-size life stages and species, fixed stations and coded tags for tracking large samples of animals over long distances and large temporal scales, inexpensive PIT systems that enable mass tagging to yield population- and community-level relevant sample sizes, incorporation of sensors into electronic tags, validation of tag attachment procedures with a focus on maintaining animal welfare, incorporation of different techniques (for example, genetics, stable isotopes) and peripheral technologies (for example, geographic information systems, hydroacoustics), development of novel analytical techniques, and extensive international collaboration. Innovations are still needed in tag miniaturization, data analysis and visualization, and in tracking animals over larger spatial scales (for example, pelagic areas of lakes) and in challenging environments (for example, large dynamic floodplain systems, under ice). There seems to be a particular need for adapting various global positioning system and satellite tagging approaches to freshwater. Electronic tagging provides a mechanism to collect detailed information from imperilled animals and species that have no direct economic value. Current and future advances will continue to improve our knowledge of the natural history of aquatic animals and ecological processes in freshwater ecosystems while facilitating evidence-based resource management and conservation.

  10. Occurrence of organic wastewater compounds in drinking water, wastewater effluent, and the Big Sioux River in or near Sioux Falls, South Dakota, 2001-2004

    USGS Publications Warehouse

    Sando, Steven K.; Furlong, Edward T.; Gray, James L.; Meyer, Michael T.

    2006-01-01

    The U.S. Geological Survey (USGS) in cooperation with the city of Sioux Falls conducted several rounds of sampling to determine the occurrence of organic wastewater compounds (OWCs) in the city of Sioux Falls drinking water and waste-water effluent, and the Big Sioux River in or near Sioux Falls during August 2001 through May 2004. Water samples were collected during both base-flow and storm-runoff conditions. Water samples were collected at 8 sites, which included 4 sites upstream from the wastewater treatment plant (WWTP) discharge, 2 sites downstream from the WWTP discharge, 1 finished drinking-water site, and 1 WWTP effluent (WWE) site. A total of 125 different OWCs were analyzed for in this study using five different analytical methods. Analyses for OWCs were performed at USGS laboratories that are developing and/or refining small-concentration (less than 1 microgram per liter (ug/L)) analytical methods. The OWCs were classified into six compound classes: human pharmaceutical compounds (HPCs); human and veterinary antibiotic compounds (HVACs); major agricultural herbicides (MAHs); household, industrial,and minor agricultural compounds (HIACs); polyaromatic hydrocarbons (PAHs); and sterol compounds (SCs). Some of the compounds in the HPC, MAH, HIAC, and PAH classes are suspected of being endocrine-disrupting compounds (EDCs). Of the 125 different OWCs analyzed for in this study, 81 OWCs had one or more detections in environmental samples reported by the laboratories, and of those 81 OWCs, 63 had acceptable analytical method performance, were detected at concentrations greater than the study reporting levels, and were included in analyses and discussion related to occurrence of OWCs in drinking water, wastewater effluent, and the Big Sioux River. OWCs in all compound classes were detected in water samples from sampling sites in the Sioux Falls area. For the five sampling periods when samples were collected from the Sioux Falls finished drinking water, only one OWC was detected at a concentration greater than the study reporting level (metolachlor; 0.0040 ug/L). During base-flow conditions, Big Sioux River sites upstream from the WWTP discharge had OWC contributions that primarily were from nonpoint animal or crop agriculture sources or had OWC concentrations that were minimal. The influence of the WWTP discharge on OWCs at downstream river sites during base-flow conditions ranged from minimal influence to substantial influence depending on the sampling period. During runoff conditions, OWCs at sites upstream from the WWTP discharge probably were primarily contributed by nonpoint animal and/or crop agriculture sources and possibly by stormwater runoff from nearby roads. OWCs at sites downstream from the WWTP discharge probably were contributed by sources other than the WWTP effluent discharge, such as stormwater runoff from urban and/or agriculture areas and/or resuspension of OWCs adsorbed to sediment deposited in the Big Sioux River. OWC loads generally were substantially smaller for upstream sites than downstream sites during both base-flow and runoff conditions.discharge had OWC contributions that primarily were from nonpoint animal or crop agriculture sources or had OWC concentrations that were minimal. The influence of the WWTP discharge on OWCs at downstream river sites during base-flow conditions ranged from minimal influence to substantial influence depending on the sampling period. During runoff conditions, OWCs at sites upstream from the WWTP discharge probably were primarily contributed by nonpoint animal and/or crop agriculture sources and possibly by stormwater runoff from nearby roads. OWCs at sites downstream from the WWTP discharge probably were contributed by sources other than the WWTP effluent discharge, such as stormwater runoff from urban and/or agriculture areas and/or resuspension of OWCs adsorbed to sediment deposited in the Big Sioux River. OWC loads generally were substantially smaller for

  11. Human health risks related to the consumption of foodstuffs of animal origin contaminated by bisphenol A.

    PubMed

    Gorecki, Sébastien; Bemrah, Nawel; Roudot, Alain-Claude; Marchioni, Eric; Le Bizec, Bruno; Faivre, Franck; Kadawathagedara, Manik; Botton, Jérémie; Rivière, Gilles

    2017-12-01

    Bisphenol A (BPA) is used in a wide variety of products and objects for consumers use (digital media such as CD's and DVD's, sport equipment, food and beverage containers, medical equipment). For humans, the main route of exposure to BPA is food. Based on previous estimates, almost 20% of the dietary exposure to BPA in the French population would be from food of animal origin. However, due to the use of composite samples, the source of the contamination had not been identified. Therefore, 322 individual samples of non-canned foods of animal origin were collected with the objectives of first updating the estimation of the exposure of the French population and second identifying the source of contamination of these foodstuffs using a specific analytical method. Compared to previous estimates in France, a decline in the contamination of the samples was observed, in particular with regard to meat. The estimated mean dietary exposures ranged from 0.048 to 0.050 μg (kg bw) -1 d -1 for 3-17 year children and adolescents, from 0.034 to 0.035 μg (kg bw) -1 d -1 for adults and from 0.047 to 0.049 μg (kg bw) -1 d -1 for pregnant women. The contribution of meat to total dietary exposure of pregnant women, adults and children was up to three times lower than the previous estimates. Despite this downward trend in contamination, the toxicological values were observed to have been exceeded for the population of pregnant women. With the aim of acquiring more knowledge about the origin the potential source(s) of contamination of non-canned foods of animal origin, a specific analytical method was developed to directly identify and quantify the presence of conjugated BPA (BPA-monoglucuronide, BPA-diglucuronide and sulphate forms) in 50 samples. No conjugated forms of BPAs were detected in the analysed samples, indicating clearly that BPA content in animal food was not due to metabolism but arise post mortem in food. This contamination may occur during food production. However, despite extensive sampling performed in several different shops (butcheries, supermarkets …. ) and in different conditions (fresh, prepared, frozen …), the source(s) of the contamination could not be specifically identified. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Isozyme composition of lactate dehydrogenase of rat skeletal muscles after flight in Kosmos-690 biosatellite. [Effects of radiation on lactate dehydrogenase

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

    Petrova, N.V.

    1978-01-01

    Rats in a ground-based model experiment, in which all flight conditions with the exception of weightlessness and accelerations and intact animals maintained under vivarium conditions served as a control. On the 10th day of flight and of the ground-based model experiments, the rats were exposed to 800 rad radiation for 24 h. Samples of soleus and plantaris muscles were taken for examination on the 2d and 27th days after landing and termination of the ground-based model experiment. Intact animals were sacrificed on the same days as experimental ones. Samples of muscle tissue were frozen in dry ice and stored formore » several days at a temperature of -70/sup 0/ before they were studied. This investigation of isozyme spectrum of LDH of skeletal muscles of rats from the Kosmos-690 satellite indicates that the changes in proportion of isozyme fractions of LDH on the 2d day after the flight are due to the effects of weightlessness; subsequent changes (27th day) in correlation between LDH fraction activity are related to the effects of radiation.« less

  13. Humoral and cellular immunity in chromium picolinate-supplemented lambs.

    PubMed

    Dallago, B S L; McManus, C M; Caldeira, D F; Campeche, A; Burtet, R T; Paim, T P; Gomes, E F; Branquinho, R P; Braz, S V; Louvandini, H

    2013-08-01

    The effects of oral supplementation of chromium picolinate (CrPic) on humoral and cellular immunity in sheep were investigated. Twenty-four male lambs divided into four treatments and received different dosages of CrPic: placebo (0), 0.250, 0.375, and 0.500 mg of chromium/animal/day during 84 days. The base ration was Panicum maximum cv Massai hay and concentrate. Blood samples were collected fortnightly for total and differential leukocyte counts. On days 28 and 56, the lambs were challenged with chicken ovalbumin I.M. Serum samples were collected on days 46 and 74 and subjected to an indirect enzyme-linked immunosorbent assay to measure IgG anti-ovalbumin. The cell-mediated immune response was determined by a delay-type hypersensitivity test using phytohemagglutinin. CrPic did not significantly affect humoral immunity in lambs but there was a negative effect on cellular immunity (P < 0.05) as Cr supplementation increased. Therefore, the level of Cr supplementation for lambs must be better studied to address its effect on stressed animals or the possible toxic effects of Cr on the animal itself or its immune system.

  14. Effect of spaceflight on the extracellular matrix of skeletal muscle after a crush injury

    NASA Technical Reports Server (NTRS)

    Stauber, W. T.; Fritz, V. K.; Burkovskaia, T. E.; Il'ina-Kakueva, E. I.

    1992-01-01

    The organization and composition of the extracellular matrix were studied in the crush-injured gastrocnemius muscle of rats subjected to 0 G. After 14 days of flight on Cosmos 2044, the gastrocnemius muscle was removed and evaluated by histochemical and immunohistochemical techniques from the five injured flight rodents and various earth-based treatment groups. In general, the repair process was similar in all injured muscle samples with regard to the organization of the extracellular matrix and myofibers. Small and large myofibers were present within an expanded extracellular matrix, indicative of myogenesis and muscle regeneration. In the tail-suspended animals, a more complete repair was observed with nonenlarged area of nonmuscle cells or matrix material visible. In contrast, the muscle samples from the flight animals were less well organized and contained more macrophages and blood vessels in the repair region, indicative of a delayed repair process, but did not demonstrate any chronic inflammation. Myofiber repair did vary in muscles from the different groups, being slowest in the flight animals and most complete in the tail-suspended ones.

  15. Does translocation influence physiological stress in the desert tortoise?

    USGS Publications Warehouse

    Drake, K.K.; Nussear, K.E.; Esque, T.C.; Barber, A.M.; Vittum, K.M.; Medica, P.A.; Tracy, C.R.; Hunter, K.W.

    2012-01-01

    Wildlife translocation is increasingly used to mitigate disturbances to animals or habitat due to human activities, yet little is known about the extent to which translocating animals causes stress. To understand the relationship between physiological stress and translocation, we conducted a multiyear study (2007–2009) using a population of desert tortoises (Gopherus agassizii) near Fort Irwin, California. Blood samples were collected from adult tortoises in three treatment groups (resident, translocated and control) for 1 year prior to and 2 years after translocation. Samples were analyzed by radioimmunoassay for plasma total corticosterone (CORT), a glucocorticoid hormone commonly associated with stress responses in reptiles. CORT values were analyzed in relation to potential covariates (animal sex, date, behavior, treatment, handling time, air temperature, home-range size, precipitation and annual plant production) among seasons and years. CORT values in males were higher than in females, and values for both varied monthly throughout the activity season and among years. Year and sex were strong predictors of CORT, and translocation explained little in terms of CORT. Based on these results, we conclude that translocation does not elicit a physiological stress response in desert tortoises.

  16. Development of an Analytical Procedure for the Determination of Multiclass Compounds for Forensic Veterinary Toxicology.

    PubMed

    Sell, Bartosz; Sniegocki, Tomasz; Zmudzki, Jan; Posyniak, Andrzej

    2018-04-01

    Reported here is a new analytical multiclass method based on QuEChERS technique, which has proven to be effective in diagnosing fatal poisoning cases in animals. This method has been developed for the determination of analytes in liver samples comprising rodenticides, carbamate and organophosphorus pesticides, coccidiostats and mycotoxins. The procedure entails addition of acetonitrile and sodium acetate to 2 g of homogenized liver sample. The mixture was shaken intensively and centrifuged for phase separation, which was followed by an organic phase transfer into a tube containing sorbents (PSA and C18) and magnesium sulfate, then it was centrifuged, the supernatant was filtered and analyzed by liquid chromatography tandem mass spectrometry. A validation of the procedure was performed. Repeatability variation coefficients <15% have been achieved for most of the analyzed substances. Analytical conditions allowed for a successful separation of variety of poisons with the typical screening detection limit at ≤10 μg/kg levels. The method was used to investigate more than 100 animals poisoning incidents and proved that is useful to be used in animal forensic toxicology cases.

  17. Screening method for the determination of tetracyclines and fluoroquinolones in animal drinking water by liquid chromatography with diode array detector.

    PubMed

    Patyra, E; Kowalczyk, E; Grelik, A; Przeniosło-Siwczyńska, M; Kwiatek, K

    2015-01-01

    A liquid chromatography - diode array detector (HPLC-DAD) procedure has been developed for the determination of oxytetracycline (OTC), tetracycline (TC), chlorotetracycline (CTC), doxycycline (DC), enrofloxacin (ENR), ciprofloxacin (CIP), sarafloxacin (SAR) and flumequine (FLU) residues in animal drinking water. This method was applied to animal drinking water. Solid-phase extraction (SPE) clean-up on an Oasis HLB cartridge allowed an extract suitable for liquid chromatographic analysis to be obtained. Chromatographic separation was carried out on a C18 analytical column, using gradient elution with 0.1% trifluoroacetic acid - acetonitrile - methanol at 30°C. The flow-rate was 0.7 mL/min and the eluate was analysed at 330 nm. The whole procedure was evaluated according to the requirements of the Commission Decision 2002/657/EC, determining specificity, decision limit (CCα), detection capacity (CCβ), limit of detection (LOD), limit of quantification (LOQ), precision and accuracy during validation of the method. The recoveries of TCs and FQs from spiked samples at the levels of 10, 100 and 1000 μg/L were higher than 82%. The developed method based on HPLC-DAD has been applied for the determination of four tetracyclines and four fluoroquinolones in animal drinking water samples.

  18. Assessing monkeypox virus prevalence in small mammals at the human-animal interface in the Democratic Republic of the Congo

    USGS Publications Warehouse

    Doty, Jeffrey B.; Malekani, Jean M.; Kalemba, Lem's N.; Stanley, William T.; Monroe, Benjamin P.; Nakazawa, Yoshinori J.; Mauldin, Matthew R.; Bakambana, Trésor L.; Liyandja Dja Liyandja , Tobit; Braden, Zachary; Wallace, Ryan; Malekani, Divin V.; McCollum, Andrea M.; Gallardo-Romero, Nadia; Kondas, Ashley; Peterson, A. Townsend; Osorio, Jorge E.; Rocke, Tonie E.; Karem, Kevin L.; Emerson, Ginny L.; Carroll, Darin S.

    2017-01-01

    During 2012, 2013 and 2015, we collected small mammals within 25 km of the town of Boende in Tshuapa Province, the Democratic Republic of the Congo. The prevalence of monkeypox virus (MPXV) in this area is unknown; however, cases of human infection were previously confirmed near these collection sites. Samples were collected from 353 mammals (rodents, shrews, pangolins, elephant shrews, a potamogale, and a hyrax). Some rodents and shrews were captured from houses where human monkeypox cases have recently been identified, but most were trapped in forests and agricultural areas near villages. Real-time PCR and ELISA were used to assess evidence of MPXV infection and other Orthopoxvirus (OPXV) infections in these small mammals. Seven (2.0%) of these animal samples were found to be anti-orthopoxvirus immunoglobulin G (IgG) antibody positive (six rodents: two Funisciurus spp.; one Graphiurus lorraineus; one Cricetomys emini; one Heliosciurus sp.; one Oenomys hypoxanthus, and one elephant shrew Petrodromus tetradactylus); no individuals were found positive in PCR-based assays. These results suggest that a variety of animals can be infected with OPXVs, and that epidemiology studies and educational campaigns should focus on animals that people are regularly contacting, including larger rodents used as protein sources. 

  19. Aggressive behavior and hair cortisol levels in captive Dorcas gazelles (Gazella dorcas) as animal-based welfare indicators.

    PubMed

    Salas, Marina; Temple, Déborah; Abáigar, Teresa; Cuadrado, Mariano; Delclaux, Maria; Enseñat, Conrad; Almagro, Vanessa; Martínez-Nevado, Eva; Quevedo, Miguel Ángel; Carbajal, Annaïs; Tallo-Parra, Oriol; Sabés-Alsina, Maria; Amat, Marta; Lopez-Bejar, Manel; Fernández-Bellon, Hugo; Manteca, Xavier

    2016-11-01

    Ensuring welfare in captive wild animal populations is important not only for ethical and legal reasons, but also to maintain healthy individuals and populations. An increased level of social behaviors such as aggression can reduce welfare by causing physical damage and chronic stress to animals. Recently, cortisol in hair has been advanced as a non-invasive indicator to quantify long-lasting stress in many species. The sensitivity of social behavior and hair cortisol concentration was evaluated in several groups of dorcas gazelles (Gazella dorcas). Four different groups of gazelles from three different zoos were observed and the expression of intra-specific affiliative and negative social behaviors was assessed across the different groups. Hair samples were taken from sub-groups of animals and analyzed for cortisol concentrations. Significant differences between groups of dorcas gazelles were found in frequency of negative social behavior and hair cortisol concentration. Despite the low sample size, these two parameters had a positive Spearman correlation coefficient (r s  = +0.80, P = 0.20). These results suggest that hair cortisol levels are sensitive to differences in the social structure of dorcas gazelles. Zoo Biol. 35:467-473, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  20. Assessing Monkeypox Virus Prevalence in Small Mammals at the Human-Animal Interface in the Democratic Republic of the Congo.

    PubMed

    Doty, Jeffrey B; Malekani, Jean M; Kalemba, Lem's N; Stanley, William T; Monroe, Benjamin P; Nakazawa, Yoshinori U; Mauldin, Matthew R; Bakambana, Trésor L; Liyandja Dja Liyandja, Tobit; Braden, Zachary H; Wallace, Ryan M; Malekani, Divin V; McCollum, Andrea M; Gallardo-Romero, Nadia; Kondas, Ashley; Peterson, A Townsend; Osorio, Jorge E; Rocke, Tonie E; Karem, Kevin L; Emerson, Ginny L; Carroll, Darin S

    2017-10-03

    During 2012, 2013 and 2015, we collected small mammals within 25 km of the town of Boende in Tshuapa Province, the Democratic Republic of the Congo. The prevalence of monkeypox virus (MPXV) in this area is unknown; however, cases of human infection were previously confirmed near these collection sites. Samples were collected from 353 mammals (rodents, shrews, pangolins, elephant shrews, a potamogale, and a hyrax). Some rodents and shrews were captured from houses where human monkeypox cases have recently been identified, but most were trapped in forests and agricultural areas near villages. Real-time PCR and ELISA were used to assess evidence of MPXV infection and other Orthopoxvirus (OPXV) infections in these small mammals. Seven (2.0%) of these animal samples were found to be anti-orthopoxvirus immunoglobulin G (IgG) antibody positive (six rodents: two Funisciurus spp.; one Graphiurus lorraineus ; one Cricetomys emini ; one Heliosciurus sp.; one Oenomys hypoxanthus , and one elephant shrew Petrodromus tetradactylus ); no individuals were found positive in PCR-based assays. These results suggest that a variety of animals can be infected with OPXVs, and that epidemiology studies and educational campaigns should focus on animals that people are regularly contacting, including larger rodents used as protein sources.

  1. Effective PCR detection of animal species in highly processed animal byproducts and compound feeds.

    PubMed

    Fumière, Olivier; Dubois, Marc; Baeten, Vincent; von Holst, Christoph; Berben, Gilbert

    2006-07-01

    In this paper we present a polymerase chain reaction (PCR)-based method for detecting meat and bone meal (MBM) in compound feedingstuffs. By choosing adequate DNA targets from an appropriate localisation in the genome, the real-time PCR method developed here proved to be robust to severe heat treatment of the MBM, showing high sensitivity in the detection of MBM. The method developed here permits the specific detection of processed pig and cattle materials treated at 134 degrees C in various feed matrices down to a limit of detection of about 0.1%. This technique has also been successfully applied to well-characterised MBM samples heated to as high as 141 degrees C, as well as to various blind feed samples with very low MBM contents. Finally, the method also passed several official European ring trials.

  2. Detection of Hepatozoon canis in stray dogs and cats in Bangkok, Thailand.

    PubMed

    Jittapalapong, Sathaporn; Rungphisutthipongse, Opart; Maruyama, Soichi; Schaefer, John J; Stich, Roger W

    2006-10-01

    A rapidly increasing stray animal population in Bangkok has caused concern regarding transmission of vector-borne and zoonotic diseases. The purpose of this study was to determine if stray animals in Bangkok are a potential reservoir of Hepatozoon, a genus of tick-borne parasites that has received little attention in Thailand. Blood samples were collected from stray companion animals near monasteries in 42 Bangkok metropolitan districts. Both dogs and cats were sampled from 26 districts, dogs alone from 4 districts and cats alone from 12 districts. Samples were collected from a total of 308 dogs and 300 cats. Light microscopy and an 18 S rRNA gene-based PCR assay were used to test these samples for evidence of Hepatozoon infection. Gamonts were observed in blood smears for 2.6% of dogs and 0.7% of cats by microscopy. The PCR assay detected Hepatozoon in buffy coats from 11.4% of dogs and 32.3% of cats tested. The prevalence of infection was the same between male and female dogs or cats, and PCR-positive dogs and cats were found in 36.6% and 36.8% of the districts surveyed, respectively. There was an association between the percentages of PCR-positive dogs and cats in districts where both host species were sampled. Sequences of representative amplicons were closest to those reported for H. canis. These results represent the first molecular confirmation that H. canis is indigenous to Thailand. The unexpectedly high prevalence of Hepatozoon among stray cats indicates that their role in the epizootiology of hepatozoonosis should be investigated.

  3. Potential arsenic exposures in 25 species of zoo animals living in CCA-wood enclosures.

    PubMed

    Gress, J; da Silva, E B; de Oliveira, L M; Zhao, Di; Anderson, G; Heard, D; Stuchal, L D; Ma, L Q

    2016-05-01

    Animal enclosures are often constructed from wood treated with the pesticide chromated copper arsenate (CCA), which leaches arsenic (As) into adjacent soil during normal weathering. This study evaluated potential pathways of As exposure in 25 species of zoo animals living in CCA-wood enclosures. We analyzed As speciation in complete animal foods, dislodgeable As from CCA-wood, and As levels in enclosure soils, as well as As levels in biomarkers of 9 species of crocodilians (eggs), 4 species of birds (feathers), 1 primate species (hair), and 1 porcupine species (quills). Elevated soil As in samples from 17 enclosures was observed at 1.0-110mg/kg, and enclosures housing threatened and endangered species had As levels higher than USEPA's risk-based Eco-SSL for birds and mammals of 43 and 46mg/kg. Wipe samples of CCA-wood on which primates sit had dislodgeable As residues of 4.6-111μg/100cm(2), typical of unsealed CCA-wood. Inorganic As doses from animal foods were estimated at 0.22-7.8μg/kg bw/d. Some As levels in bird feathers and crocodilian eggs were higher than prior studies on wild species. However, hair from marmosets had 6.37mg/kg As, 30-fold greater than the reference value, possibly due to their inability to methylate inorganic As. Our data suggested that elevated As in soils and dislodgeable As from CCA-wood could be important sources of As exposure for zoo animals. Published by Elsevier B.V.

  4. Direct Detection of Soil-Bound Prions

    PubMed Central

    Genovesi, Sacha; Leita, Liviana; Sequi, Paolo; Andrighetto, Igino; Sorgato, M. Catia; Bertoli, Alessandro

    2007-01-01

    Scrapie and chronic wasting disease are contagious prion diseases affecting sheep and cervids, respectively. Studies have indicated that horizontal transmission is important in sustaining these epidemics, and that environmental contamination plays an important role in this. In the perspective of detecting prions in soil samples from the field by more direct methods than animal-based bioassays, we have developed a novel immuno-based approach that visualises in situ the major component (PrPSc) of prions sorbed onto agricultural soil particles. Importantly, the protocol needs no extraction of the protein from soil. Using a cell-based assay of infectivity, we also report that samples of agricultural soil, or quartz sand, acquire prion infectivity after exposure to whole brain homogenates from prion-infected mice. Our data provide further support to the notion that prion-exposed soils retain infectivity, as recently determined in Syrian hamsters intracerebrally or orally challanged with contaminated soils. The cell approach of the potential infectivity of contaminated soil is faster and cheaper than classical animal-based bioassays. Although it suffers from limitations, e.g. it can currently test only a few mouse prion strains, the cell model can nevertheless be applied in its present form to understand how soil composition influences infectivity, and to test prion-inactivating procedures. PMID:17957252

  5. Prevalence and characterization of Salmonella enterica from the feces of cattle, poultry, swine and hedgehogs in Burkina Faso and their comparison to human Salmonella isolates

    PubMed Central

    2013-01-01

    Background Production and wild animals are major sources of human salmonellosis and animals raised for food also play an important role in transmission of antimicrobial resistant Salmonella strains to humans. Furthermore, in sub-Saharan Africa non-typhoidal Salmonella serotypes are common bloodstream isolates in febrile patients. Yet, little is known about the environmental reservoirs and predominant modes of transmission of these pathogens. The purpose of this study was to discover potential sources and distribution vehicles of Salmonella by isolating strains from apparently healthy slaughtered food animals and wild hedgehogs and by determining the genetic relatedness between the strains and human isolates. For this purpose, 729 feces samples from apparently healthy slaughtered cattle (n = 304), poultry (n = 350), swine (n = 50) and hedgehogs (n = 25) were examined for the presence of Salmonella enterica in Burkina Faso. The isolates were characterized by serotyping, antimicrobial-susceptibility testing, phage typing, and pulsed-field gel electrophoresis (PFGE) with XbaI and BlnI restriction enzymes. Results Of the 729 feces samples, 383 (53%) contained Salmonella, representing a total of 81 different serotypes. Salmonella was present in 52% of the cattle, 55% of the poultry, 16% of the swine and 96% of the hedgehog feces samples. Antimicrobial resistance was detected in 14% of the isolates. S. Typhimurium isolates from poultry and humans (obtained from a previous study) were multiresistant to the same antimicrobials (ampicillin, chloramphenicol, streptomycin, sulfonamides and trimethoprim), had the same phage type DT 56 and were closely related in PFGE. S. Muenster isolates from hedgehogs had similar PFGE patterns as the domestic animals. Conclusions Based on our results it seems that production and wild animals can share the same Salmonella serotypes and potentially transmit some of them to humans. As the humans and animals often live in close vicinity in Africa and the hygiene control of the meat retail chain is defective, high Salmonella carriage rates of the animals can pose a major public health risk in Burkina Faso. This underlines the necessity for a joint and coordinated surveillance and monitoring programs for salmonellosis in Africa. PMID:24215206

  6. Microscopic and Molecular Detection of Theileria (Babesia) Equi Infection in Equids of Kurdistan Province, Iran

    PubMed Central

    HABIBI, Gholamreza; ESMAEILNIA, Kasra; HABLOLVARID, Mohammad Hasan; AFSHARI, Asghar; ZAMEN, Mohsen; BOZORGI, Soghra

    2016-01-01

    Background: Equine piroplasmosis (EP) is the cause of persistent tick-borne infection with no symptoms, but the most important problem of EP is due to the persistent carrier state. Carrier animals to Babesia (Theileria) equi (Laveran 1901) and B. caballi (Nuttall, 1910) infestation could be identified by extremely sensitive PCR-based method. The purpose of this study was to identify the causative agents of equine piroplasmosis based on molecular and microscopic assays in equids from Kurdistan Province, Iran. Methods: Thirty one horse and mule blood samples were used with history of living in Kurdistan Province of Iran. The blood specimens were utilized for T. equi and B. caballi DNA identification by PCR and Giemsa stained smears for microscopic observation. Results: The results clearly showed the presence of B. (Theileria) equi DNA in 30 of 31 blood samples (96.77%), but the microscopic examination revealed the 3 of 31 positive Babesia like organisms in the red blood cells (9.67%). Conclusion: The obtained results demonstrated the presence of hidden B. (Theileria) equi infection in horses with previous habitance in Kurdistan Province of Iran. The carrier animals became a main source of infection and can transmit the disease. Therefore, hidden infection might be considered as a health threatening and limiting factor in animals used in therapeutic antisera research and production centers. PMID:27095973

  7. Multiple detection of zoonotic variegated squirrel bornavirus 1 RNA in different squirrel species suggests a possible unknown origin for the virus.

    PubMed

    Schlottau, Kore; Hoffmann, Bernd; Homeier-Bachmann, Timo; Fast, Christine; Ulrich, Rainer G; Beer, Martin; Hoffmann, Donata

    2017-09-01

    The recently discovered variegated squirrel bornavirus 1 (VSBV-1) caused the death of three squirrel breeders in Germany. Subsequent first screening of squirrels with in vivo collected swab samples and a VSBV-1-specific RT-qPCR revealed not only variegated squirrel infections (Sciurus variegatoides), but also Prevost's squirrels (Callosciurus prevostii) as positive for VSBV-1 genome. In this study, 328 squirrels were tested using the established RT-qPCR assays. In 16 individual animals VSBV-1 RNA could be detected; 15 individuals were from small breedings and zoological gardens in Germany, with the remaining individual being from a zoological garden in Croatia. Positive animals belonged to the species C. prevostii, C. finlaysonii, and Tamiops swinhoei within the subfamily Callosciurinae and Sciurus granatensis within the subfamily Sciurinae. Repeated non-invasive oral swab sampling in one holding indicated positive animals months after a first negative result. Besides the oral swabs, VSBV-1 was also detected in fecal (pool) samples allowing the future monitoring of squirrel holdings based on RT-qPCR investigation of such samples. The detection in zoological gardens emphasizes the need for further investigations into the transmission route to humans in order to develop rational public health measures for prevention of transmission. Finally, the detection of several closely related VSBV-1 sequences in squirrels from different subfamilies raises questions as to the origin of the virus.

  8. Nucleic acid purification from plants, animals and microbes in under 30 seconds

    PubMed Central

    Zou, Yiping; Wang, Yuling; Wee, Eugene; Turni, Conny; Blackall, Patrick J.; Trau, Matt; Botella, Jose Ramon

    2017-01-01

    Nucleic acid amplification is a powerful molecular biology tool, although its use outside the modern laboratory environment is limited due to the relatively cumbersome methods required to extract nucleic acids from biological samples. To address this issue, we investigated a variety of materials for their suitability for nucleic acid capture and purification. We report here that untreated cellulose-based paper can rapidly capture nucleic acids within seconds and retain them during a single washing step, while contaminants present in complex biological samples are quickly removed. Building on this knowledge, we have successfully created an equipment-free nucleic acid extraction dipstick methodology that can obtain amplification-ready DNA and RNA from plants, animals, and microbes from difficult biological samples such as blood and leaves from adult trees in less than 30 seconds. The simplicity and speed of this method as well as the low cost and availability of suitable materials (e.g., common paper towelling), means that nucleic acid extraction is now more accessible and affordable for researchers and the broader community. Furthermore, when combined with recent advancements in isothermal amplification and naked eye DNA visualization techniques, the dipstick extraction technology makes performing molecular diagnostic assays achievable in limited resource settings including university and high school classrooms, field-based environments, and developing countries. PMID:29161268

  9. Uncovering a latent multinomial: Analysis of mark-recapture data with misidentification

    USGS Publications Warehouse

    Link, W.A.; Yoshizaki, J.; Bailey, L.L.; Pollock, K.H.

    2010-01-01

    Natural tags based on DNA fingerprints or natural features of animals are now becoming very widely used in wildlife population biology. However, classic capture-recapture models do not allow for misidentification of animals which is a potentially very serious problem with natural tags. Statistical analysis of misidentification processes is extremely difficult using traditional likelihood methods but is easily handled using Bayesian methods. We present a general framework for Bayesian analysis of categorical data arising from a latent multinomial distribution. Although our work is motivated by a specific model for misidentification in closed population capture-recapture analyses, with crucial assumptions which may not always be appropriate, the methods we develop extend naturally to a variety of other models with similar structure. Suppose that observed frequencies f are a known linear transformation f = A???x of a latent multinomial variable x with cell probability vector ?? = ??(??). Given that full conditional distributions [?? | x] can be sampled, implementation of Gibbs sampling requires only that we can sample from the full conditional distribution [x | f, ??], which is made possible by knowledge of the null space of A???. We illustrate the approach using two data sets with individual misidentification, one simulated, the other summarizing recapture data for salamanders based on natural marks. ?? 2009, The International Biometric Society.

  10. Uncovering a Latent Multinomial: Analysis of Mark-Recapture Data with Misidentification

    USGS Publications Warehouse

    Link, W.A.; Yoshizaki, J.; Bailey, L.L.; Pollock, K.H.

    2009-01-01

    Natural tags based on DNA fingerprints or natural features of animals are now becoming very widely used in wildlife population biology. However, classic capture-recapture models do not allow for misidentification of animals which is a potentially very serious problem with natural tags. Statistical analysis of misidentification processes is extremely difficult using traditional likelihood methods but is easily handled using Bayesian methods. We present a general framework for Bayesian analysis of categorical data arising from a latent multinomial distribution. Although our work is motivated by a specific model for misidentification in closed population capture-recapture analyses, with crucial assumptions which may not always be appropriate, the methods we develop extend naturally to a variety of other models with similar structure. Suppose that observed frequencies f are a known linear transformation f=A'x of a latent multinomial variable x with cell probability vector pi= pi(theta). Given that full conditional distributions [theta | x] can be sampled, implementation of Gibbs sampling requires only that we can sample from the full conditional distribution [x | f, theta], which is made possible by knowledge of the null space of A'. We illustrate the approach using two data sets with individual misidentification, one simulated, the other summarizing recapture data for salamanders based on natural marks.

  11. Food animal transport: a potential source of community exposures to health hazards from industrial farming (CAFOs).

    PubMed

    Rule, Ana M; Evans, Sean L; Silbergeld, Ellen K

    2008-01-01

    Use of antimicrobial feed additives in food animal production is associated with selection for drug resistance in bacterial pathogens, which can then be released into the environment through occupational exposures, high volume ventilation of animal houses, and land application of animal wastes. We tested the hypothesis that current methods of transporting food animals from farms to slaughterhouses may result in pathogen releases and potential exposures of persons in vehicles traveling on the same road. Air and surface samples were taken from cars driving behind poultry trucks for 17 miles. Air conditioners and fans were turned off and windows fully opened. Background and blank samples were used for quality control. Samples were analyzed for susceptible and drug-resistant strains. Results indicate an increase in the number of total aerobic bacteria including both susceptible and drug-resistant enterococci isolated from air and surface samples, and suggest that food animal transport in open crates introduces a novel route of exposure to harmful microorganisms and may disseminate these pathogens into the general environment. These findings support the need for further exposure characterization, and attention to improving methods of food animal transport, especially in highly trafficked regions of high density farming such as the Delmarva Peninsula.

  12. An analysis of human pathogens found in horse/mule manure along the John Muir Trail in Kings Canyon and Sequoia and Yosemite National Parks.

    PubMed

    Derlet, Robert Wayne; Carlson, James Reynolds

    2002-01-01

    To determine the prevalence of microorganisms that are potentially pathogenic for humans in horse/mule manure along the John Muir Trail (JMT). Random samples of horse/mule manure were collected along sections of the JMT in Yosemite, Kings Canyon, and Sequoia national parks (NP), as well as in portions of the Pacific Crest Trail (PCT) and selected JMT/PCT access trails. Convenience samples of wild animal scat found within I mile of trails were also collected. The fresh specimens were individually preserved both in 0.9% saline and polyvinyl alcohol (PVA)-containing tubes and stored at 4 degrees C until time of analysis. Bacteriological analysis was performed using standard microbiology laboratory procedures. PVA samples were stained with trichrome and were then examined by a parasitologist. Collection: A total of 186 trail miles were sampled, including 113 on the JMT (Yosemite 37, Kings 53, and Sequoia 23). The PCT samplings included 24 miles, and NP and wilderness area access trails added an additional 49 miles. A total of 102 samples were collected, which included 81 samples from pack animals and 21 identified as having come from wild animals. Pack Animal Bacteria: All plated specimens grew large numbers of commensal gut flora. Potential pathogenic bacteria were found in only 12 samples and included Hafnia alvei (4), Serratia odorifera (1), Citrobacter freundii (1), Escherichia vulneris (1), Clostridium clostridioforme (1), Yersinia enterocolitica (1), Sherwinella putraformus (1), and Enterobacter spp (4). No Escherichia coli O157, Salmonella, or Aeromonas were found. Microscopic examination for protozoal organisms revealed occasional commensal ciliates and I Giardia. Wild Animal Pathogens: One specimen grew Y enterocolitica, and another grew Enterobacter amnigenus. We found a low prevalence of human pathogens in pack animal manure on the JMT.

  13. Seroprevalence Survey for Microsporidia in Common Bottlenose Dolphin ( Tursiops truncatus): Example of A Quantitative Approach Based on Immunoblotting.

    PubMed

    Desoubeaux, Guillaume; Peschke, Roman; Le-Bert, Carolina; Fravel, Vanessa; Soto, Jeny; Jensen, Eric D; Flower, Jennifer E; Wells, Randall; Joachim, Anja; Cray, Carolyn

    2018-05-09

    Little is known about microsporidiosis pathogenicity in cetaceans. Here we report seroprevalence of 76% for microsporidia in blood samples from common bottlenose dolphins ( Tursiops truncatus), from animals managed under human care ( n=108) or captured for health assessments ( n=13) and released.

  14. Screening Complex Effluents for Estrogenic Activity with the T47D-Kbluc Cell Bioassay: Assay Optimization and Comparison to In Vivo Responses in Fish

    EPA Science Inventory

    The endocrine activity of complex mixtures of chemicals associated with wastewater treatment plant effluents, runoff from concentrated animal feeding operations (CAFOs), and/or other environmental samples can be difficult to characterize based on analytical chemistry. In vitro bi...

  15. Cost effective raspberry pi-based radio frequency identification tagging of mice suitable for automated in vivo imaging.

    PubMed

    Bolaños, Federico; LeDue, Jeff M; Murphy, Timothy H

    2017-01-30

    Automation of animal experimentation improves consistency, reduces potential for error while decreasing animal stress and increasing well-being. Radio frequency identification (RFID) tagging can identify individual mice in group housing environments enabling animal-specific tracking of physiological parameters. We describe a simple protocol to radio frequency identification (RFID) tag and detect mice. RFID tags were injected sub-cutaneously after brief isoflurane anesthesia and do not require surgical steps such as suturing or incisions. We employ glass-encapsulated 125kHz tags that can be read within 30.2±2.4mm of the antenna. A raspberry pi single board computer and tag reader enable automated logging and cross platform support is possible through Python. We provide sample software written in Python to provide a flexible and cost effective system for logging the weights of multiple mice in relation to pre-defined targets. The sample software can serve as the basis of any behavioral or physiological task where users will need to identify and track specific animals. Recently, we have applied this system of tagging to automated mouse brain imaging within home-cages. We provide a cost effective solution employing open source software to facilitate adoption in applications such as automated imaging or tracking individual animal weights during tasks where food or water restriction is employed as motivation for a specific behavior. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Molecular characterization of Blastocystis isolates from zoo animals and their animal-keepers.

    PubMed

    Parkar, Unaiza; Traub, Rebecca J; Vitali, Simone; Elliot, Aileen; Levecke, Bruno; Robertson, Ian; Geurden, Thomas; Steele, Jan; Drake, Bev; Thompson, R C Andrew

    2010-04-19

    Blastocystis is an enteric protist and one of the most frequently reported parasitic infections in humans and a variety of animal hosts. It has also been reported in numerous parasite surveys of animals in zoological gardens and in particular in non-human primate species. PCR-based methods capable of the direct detection of Blastocystis in faeces were used to detect Blastocystis from various hosts, including non-human primates, Australian native fauna, elephants and giraffes, as well as their keepers from a Western Australian zoo. Additional faecal samples were also collected from elephants and giraffes from four other zoos in Amsterdam (The Netherlands), Antwerp (Belgium), Melbourne and Werribee (Australia). Information regarding the general health and lifestyle of the human volunteers were obtained by questionnaire. Overall, 42% and 63% of animals and zoo-keepers sampled from the Western Australian zoo were positive for Blastocystis, respectively. The occurrence of Blastocystis in elephants and giraffes from other cities was similar. This is the first report of Blastocystis found in the elephant, giraffe, quokka, southern hairy nosed wombat and western grey kangaroo. Three novel and what appear to be highly host-specific subtypes (STs) of Blastocystis in the elephant, giraffe and quokka are also described. These findings indicate that further exploration of the genetic diversity of Blastocystis is crucial. Most zoo-keepers at the Perth Zoo were harbouring Blastocystis. Four of these zoo-keeper isolates were identical to the isolates from the southern hairy nosed wombat and five primate species.

  17. Multiresidue method for pesticide residue analysis in food of animal and plant origin based on GC or LC and MS or MS/MS.

    PubMed

    Muñoz, Eva; Muñoz, Gloria; Pineda, Laura; Serrahima, Eulalia; Centrich, Francesc

    2012-01-01

    A multiresidue method based on GC or LC and MS or MS/MS for the determination of 204 pesticides in diverse food matrixes of animal and plant origin is described. The method can include different stages of cleanup according to the chemical characteristics of each sample. Samples were extracted using accelerated solvent extraction. Those with a high fat content or that contained chlorophyll required further purification by gel permeation chromatography and/or SPE (ENVI-Carb). The methodology developed here was fully validated; the LOQs for the 204 pesticides are presented. The LOQ values lie between 0.01 to 0.02 mg/kg. However, in some cases, mainly in baby food, they were as low as 0.003 mg/kg, thereby meeting European Union requirements on maximum residue levels for pesticides, as outlined in European regulation 396/2005 and the Commission Directive 2003/13/EC. The procedure has been accredited for a wide scope of pesticides and matrixes by the Spanish Accreditation Body (ENAC) following ISO/IEC 17025:2005, as outlined in ENAC technical note NT-19.

  18. The usefulness of direct agglutination test, enzyme-linked immunosorbent assay and polymerase chain reaction for the detection of Toxoplasma gondii in wild animals.

    PubMed

    Kornacka, Aleksandra; Cybulska, Aleksandra; Bień, Justyna; Goździk, Katarzyna; Moskwa, Bożena

    2016-09-15

    The aim of the study was to compare the usefulness of two antibody-based methods, the direct agglutination test (DAT) and enzyme linked immuosorbent assay (ELISA), with that of the polymerase chain reaction (PCR) for detecting anti-Toxoplasma gondii in samples derived from naturally-infected wild animals. Antibodies against T. gondii were detected in meat juice samples collected from 129 free- living carnivores and omnivores. T. gondii seroprevalence was confirmed in 73,6% of examined samples when DAT and ELISA were used separately, but in only 88,4% samples when both immunological tests were used in parallel. PCR results confirmed the presence of DNA of the parasite in 24 of all the 129 samples. Sixteen samples were classified as positive when all three tests were used. A moderate degree of agreement was found between DAT and ELISA (κ=0.55). However, no agreement was found between the molecular and serological tests: κ=-1.75 for DAT versus PCR; κ=-1.67 ELISA versus PCR. By using both serological tests, antibodies against T. gondii were found in 77.5% of red foxes, 12.5% of badgers, 40% of martens and 8.3% of raccoon dogs. Antibodies against the parasite were detected also in one mink, but not in the sample derived from a polecat. T.gondii DNA was found in the brain tissue of 20 red foxes, three badgers and one raccoon dog. Our studies confirm that ELISA and DAT are suitable and reliable techniques for T. gondii antibody detection in meat juice from wild animals when serum samples are unavailable. Positive results obtained by immunological tests do not always reflect that the host was infected by T. gondii. They indicate only a contact with parasite. PCR should be used to confirm te presence of DNA from T. gondii. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Host Specificity and Source of Enterocytozoon bieneusi Genotypes in a Drinking Source Watershed

    PubMed Central

    Guo, Yaqiong; Alderisio, Kerri A.; Yang, Wenli; Cama, Vitaliano; Xiao, Lihua

    2014-01-01

    To assess the host specificity of Enterocytozoon bieneusi and to track the sources of E. bieneusi contamination, we genotyped E. bieneusi in wildlife and stormwater from the watershed of New York City's source water, using ribosomal internal transcribed spacer (ITS)-based PCR and sequence analyses. A total of 255 specimens from 23 species of wild mammals and 67 samples from stormwater were analyzed. Seventy-four (29.0%) of the wildlife specimens and 39 (58.2%) of the stormwater samples from streams were PCR positive. Altogether, 20 E. bieneusi genotypes were found, including 8 known genotypes and 12 new ones. Sixteen and five of the genotypes were seen in animals and stormwater from the watershed, respectively, with WL4 being the most common genotype in both animals (35 samples) and stormwater (23 samples). The 20 E. bieneusi genotypes belonged to five genogroups (groups 1, 3, 4, and 7 and an outlier), with only 23/113 (20.4%) E. bieneusi-positive samples belonging to zoonotic genogroup 1 and 3/20 genotypes ever being detected in humans. The two genogroups previously considered host specific, groups 3 and 4, were both detected in multiple groups of mammals. Thus, with the exception of the type IV, Peru11, and D genotypes, which were detected in only 7, 5, and 2 animals, respectively, most E. bieneusi strains in most wildlife samples and all stormwater samples in the watershed had no known public health significance, as these types have not previously been detected in humans. The role of different species of wild mammals in the contribution of E. bieneusi contamination in stormwater was supported by determinations of host-adapted Cryptosporidium species/genotypes in the same water samples. Data from this study indicate that the host specificity of E. bieneusi group 3 is broader than originally thought, and wildlife is the main source of E. bieneusi in stormwater in the watershed. PMID:24141128

  20. 9 CFR 327.11 - Receipts to importers for import product samples.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... product samples. 327.11 Section 327.11 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE... laboratory examination are to be used exclusively for that purpose, official receipts shall be issued and... collected, date of collection, and that the sample was collected for laboratory examination. The duplicate...

  1. Evaluation of environmental sampling methods for detection of Salmonella enterica in a large animal veterinary hospital.

    PubMed

    Goeman, Valerie R; Tinkler, Stacy H; Hammac, G Kenitra; Ruple, Audrey

    2018-04-01

    Environmental surveillance for Salmonella enterica can be used for early detection of contamination; thus routine sampling is an integral component of infection control programs in hospital environments. At the Purdue University Veterinary Teaching Hospital (PUVTH), the technique regularly employed in the large animal hospital for sample collection uses sterile gauze sponges for environmental sampling, which has proven labor-intensive and time-consuming. Alternative sampling methods use Swiffer brand electrostatic wipes for environmental sample collection, which are reportedly effective and efficient. It was hypothesized that use of Swiffer wipes for sample collection would be more efficient and less costly than the use of gauze sponges. A head-to-head comparison between the 2 sampling methods was conducted in the PUVTH large animal hospital and relative agreement, cost-effectiveness, and sampling efficiency were compared. There was fair agreement in culture results between the 2 sampling methods, but Swiffer wipes required less time and less physical effort to collect samples and were more cost-effective.

  2. Importance of cystic echinococcosis in slaughtered herbivores from Iran.

    PubMed

    Ezatpour, Behrouz; Farhadi, Seid Jafar; Azami, Mehdi; Alirezaei, Masoud; Ebrahimzadeh, Farzad

    2015-06-01

    Cystic echinococcosis (CE) or hydatidosis is well-known as one of the zoonotic diseases in world-wide including Iran. Hydatidosis was considered as a disease that causes severe reduction in meat wool and milk in livestock animals which all of them indicating its importance. Thus, present study was designed to evaluated prevalence of CE in slaughtered animals from Delfan region in Lorestan province of Iran. The samples 6,885 animals were considered based on type of species in a slaughter from Lorestan province. The study performed from 3 April 2009 to 3 April 2012 and inspection carried out from 4,101 cattle, 2,150 sheep and 634 goat. The liver and lungs examined based on CE and showed the highest prevalence in cattle (25.7 %) and the lowest 3.8 % in goat, likewise, CE was more in the lung than to liver. There was significant difference between species of animals and infected organ (P < 0.001). The highest prevalence was seen in winter (32.8 % for cattle, 8.1 % for goat) while, it was 15.7 % for sheep in summer (P = 0.04). Overall these data indicate the necessity of disease control strategy for reduction of CE.

  3. Real-Time Microfluidic Blood-Counting System for PET and SPECT Preclinical Pharmacokinetic Studies.

    PubMed

    Convert, Laurence; Lebel, Réjean; Gascon, Suzanne; Fontaine, Réjean; Pratte, Jean-François; Charette, Paul; Aimez, Vincent; Lecomte, Roger

    2016-09-01

    Small-animal nuclear imaging modalities have become essential tools in the development process of new drugs, diagnostic procedures, and therapies. Quantification of metabolic or physiologic parameters is based on pharmacokinetic modeling of radiotracer biodistribution, which requires the blood input function in addition to tissue images. Such measurements are challenging in small animals because of their small blood volume. In this work, we propose a microfluidic counting system to monitor rodent blood radioactivity in real time, with high efficiency and small detection volume (∼1 μL). A microfluidic channel is built directly above unpackaged p-i-n photodiodes to detect β-particles with maximum efficiency. The device is embedded in a compact system comprising dedicated electronics, shielding, and pumping unit controlled by custom firmware to enable measurements next to small-animal scanners. Data corrections required to use the input function in pharmacokinetic models were established using calibrated solutions of the most common PET and SPECT radiotracers. Sensitivity, dead time, propagation delay, dispersion, background sensitivity, and the effect of sample temperature were characterized. The system was tested for pharmacokinetic studies in mice by quantifying myocardial perfusion and oxygen consumption with (11)C-acetate (PET) and by measuring the arterial input function using (99m)TcO4 (-) (SPECT). Sensitivity for PET isotopes reached 20%-47%, a 2- to 10-fold improvement relative to conventional catheter-based geometries. Furthermore, the system detected (99m)Tc-based SPECT tracers with an efficiency of 4%, an outcome not possible through a catheter. Correction for dead time was found to be unnecessary for small-animal experiments, whereas propagation delay and dispersion within the microfluidic channel were accurately corrected. Background activity and sample temperature were shown to have no influence on measurements. Finally, the system was successfully used in animal studies. A fully operational microfluidic blood-counting system for preclinical pharmacokinetic studies was developed. Microfluidics enabled reliable and high-efficiency measurement of the blood concentration of most common PET and SPECT radiotracers with high temporal resolution in small blood volume. © 2016 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

  4. Difference between beta1-adrenoceptor autoantibodies of human and animal origin-Limitations detecting beta1-adrenoceptor autoantibodies using peptide based ELISA technology.

    PubMed

    Wenzel, Katrin; Schulze-Rothe, Sarah; Müller, Johannes; Wallukat, Gerd; Haberland, Annekathrin

    2018-01-01

    Cell-based analytics for the detection of the beta1-adrenoceptor autoantibody (beta1-AAB) are functional, yet difficult to handle, and should be replaced by easily applicable, routine lab methods. Endeavors to develop solid-phase-based assays such as ELISA to exploit epitope moieties for trapping autoantibodies are ongoing. These solid-phase-based assays, however, are often unreliable when used with human patient material, in contrast to animal derived autoantibodies. We therefore tested an immunogen peptide-based ELISA for the detection of beta1-AAB, and compared commercially available goat antibodies against the 2nd extracellular loop of human beta1-adrenoceptor (ADRB1-AB) to autoantibodies enriched from patient material. The functionality of these autoantibodies was tested in a cell based assay for comparison and their structural appearance was investigated using 2D gel electrophoresis. The ELISA showed a limit of detection for ADRB1-AB of about 1.5 nmol antibody/L when spiked in human control serum and only about 25 nmol/L when spiked in species identical (goat) matrix material. When applied to samples of human origin, the ELISA failed to identify the specific beta1-AABs. A low concentration of beta1-AAB, together with structural inconsistency of the patient originated samples as seen from the 2D Gel appearance, might contribute to the failure of the peptide based ELISA technology to detect human beta1-AABs.

  5. Two models of the sound-signal frequency dependence on the animal body size as exemplified by the ground squirrels of Eurasia (mammalia, rodentia).

    PubMed

    Nikol'skii, A A

    2017-11-01

    Dependence of the sound-signal frequency on the animal body length was studied in 14 ground squirrel species (genus Spermophilus) of Eurasia. Regression analysis of the total sample yielded a low determination coefficient (R 2 = 26%), because the total sample proved to be heterogeneous in terms of signal frequency within the dimension classes of animals. When the total sample was divided into two groups according to signal frequency, two statistically significant models (regression equations) were obtained in which signal frequency depended on the body size at high determination coefficients (R 2 = 73 and 94% versus 26% for the total sample). Thus, the problem of correlation between animal body size and the frequency of their vocal signals does not have a unique solution.

  6. Animal cruelty by children exposed to domestic violence.

    PubMed

    Currie, Cheryl L

    2006-04-01

    The first objective of this study was to determine if children exposed to domestic violence were significantly more likely to be cruel to animals than children not exposed to violence. The second was to determine if there were significant age and gender differences between children who were and were not cruel to animals. A community sample of 47 mothers with two children and a history of domestic violence were compared to a matched sample of 45 mothers with two children who did not have such a history. Children exposed to domestic violence were significantly more likely to have been cruel to animals than children not exposed to violence. The age and gender of children who were cruel to animals did not differ from children who were not cruel to animals. However, exposed children cruel to animals were significantly older than non-exposed children cruel to animals. Animal cruelty by children is correlated with exposure to domestic violence.

  7. Risk-Based Immunization Policies and Tuberculosis Screening Practices for Animal Care and Research Workers in the United States: Survey Results and Recommendations

    PubMed Central

    Weigler, Benjamin J; Cooper, Donna R; Hankenson, F Claire

    2012-01-01

    A national survey was conducted to assess immunization practices and tuberculosis screening methods for animal care and research workers in biomedical settings throughout the United States. Veterinarians (n = 953) were surveyed via a web-based mechanism; completed surveys (n = 308) were analyzed. Results showed that occupational health and safety programs were well-developed, enrolling veterinary, husbandry, and research staff at rates exceeding 90% and involving multiple modalities of health assessments and risk communication for vaccine-preventable diseases. Most (72.7%) institutions did not store serum samples from animal research personnel. More than half of the institutions housed nonhuman primates and maintained tuberculosis screening programs, although screening methods varied. Immunization protocols included various recommended or required vaccines that differed depending on job duties, type of institution, and nature of scientific programs. A single case of an identified vaccine–preventable illness in a laboratory worker was noted. Tetanus toxoid was the predominant vaccine administered (91.7%) to animal care and research workers, followed by hepatitis B (54.8%), influenza (39.9%), and rabies (38.3%). For some immunization protocols, an inconsistent rationale for administration was evident. Indications that animal care and research workers are unprotected from work-related etiologic agents did not emerge from this survey; rather, existing guidelines from the Advisory Committee on Immunization Practices and available biologics seem sufficient to address most needs of the laboratory animal research community. Institutions should commit to performance-based standards in parallel with context-specific risk assessment methods to maintain occupational health and safety programs and practices appropriate to their needs. PMID:23312084

  8. Potential Information Loss Due to Categorization of Minimum Inhibitory Concentration Frequency Distributions.

    PubMed

    Mazloom, Reza; Jaberi-Douraki, Majid; Comer, Jeffrey R; Volkova, Victoriya

    2018-01-01

    A bacterial isolate's susceptibility to antimicrobial is expressed as the lowest drug concentration inhibiting its visible growth, termed minimum inhibitory concentration (MIC). The susceptibilities of isolates from a host population at a particular time vary, with isolates with specific MICs present at different frequencies. Currently, for either clinical or monitoring purposes, an isolate is most often categorized as Susceptible, Intermediate, or Resistant to the antimicrobial by comparing its MIC to a breakpoint value. Such data categorizations are known in statistics to cause information loss compared to analyzing the underlying frequency distributions. The U.S. National Antimicrobial Resistance Monitoring System (NARMS) includes foodborne bacteria at the food animal processing and retail product points. The breakpoints used to interpret the MIC values for foodborne bacteria are those relevant to clinical treatments by the antimicrobials in humans in whom the isolates were to cause infection. However, conceptually different objectives arise when inference is sought concerning changes in susceptibility/resistance across isolates of a bacterial species in host populations among different sampling points or times. For the NARMS 1996-2013 data for animal processing and retail, we determined the fraction of comparisons of susceptibility/resistance to 44 antimicrobial drugs of twelve classes of a bacterial species in a given animal host or product population where there was a significant change in the MIC frequency distributions between consecutive years or the two sampling points, while the categorization-based analyses concluded no change. The categorization-based analyses missed significant changes in 54% of the year-to-year comparisons and in 71% of the slaughter-to-retail within-year comparisons. Hence, analyses using the breakpoint-based categorizations of the MIC data may miss significant developments in the resistance distributions between the sampling points or times. Methods considering the MIC frequency distributions in their entirety may be superior for epidemiological analyses of resistance dynamics in populations.

  9. Determination of melamine in animal feed based on liquid chromatography tandem mass spectrometry analysis and dynamic microwave-assisted extraction coupled on-line with strong cation-exchange resin clean-up.

    PubMed

    Chen, Ligang; Zeng, Qinglei; Du, Xiaobo; Sun, Xin; Zhang, Xiaopan; Xu, Yang; Yu, Aimin; Zhang, Hanqi; Ding, Lan

    2009-11-01

    In this work, a new method was developed for the determination of melamine (MEL) in animal feed. The method was based on the on-line coupling of dynamic microwave-assisted extraction (DMAE) to strong cation-exchange (SCX) resin clean-up. The MEL was first extracted by 90% acidified methanol aqueous solution (v/v, pH = 3) under the action of microwave energy, and then the extract was cooled and passed through the SCX resin. Thus, the protonated MEL was retained on the resin through ion exchange interaction and the sample matrixes were washed out. Some obvious benefits were achieved, such as acceleration of analytical process, together with reduction in manual handling, risk of contamination, loss of analyte, and sample consumption. Finally, the analyte was separated by a liquid chromatograph with a SCX analytical column, and then identified and quantitatived by a tandem mass spectrometry with positive ionization mode and multiple-reaction monitoring. The DMAE parameters were optimized by the Box-Behnken design. The linearity of quantification obtained by analyzing matrix-matched standards is in the range of 50-5,000 ng g(-1). The limit of detection and limit of quantification obtained are 12.3 and 41.0 ng g(-1), respectively. The mean intra- and inter-day precisions expressed as relative standard deviations with three fortified levels (50, 250, and 500 ng g(-1)) are 5.1% and 7.3%, respectively, and the recoveries of MEL are in the range of 76.1-93.5%. The proposed method was successfully applied to determine MEL in different animal feeds obtained from the local market. MEL was detectable with the contents of 279, 136, and 742 ng g(-1) in three samples.

  10. Leptospira spp. infection in sheep herds in southeast Brazil

    PubMed Central

    2014-01-01

    Background With the aim of studying Leptospira spp. infection in sheep herds, blood samples and respective kidney and liver fragments were collected from 100 animals from twenty different properties during slaughter at a meat company in the Sorocaba region, São Paulo state, southeast Brazil. The microscopic agglutination test (MAT) was performed with 29 strains of Leptospira spp. To identify the agent in the liver and kidney, 100 samples of each tissue were submitted to culture in Fletcher medium and analyzed by the polymerase chain reaction (PCR) for Leptospira spp. Results MAT detected 23 samples serologically positive for one or more Leptospira spp. serovars and significantly more for Autumnalis. Eight (4%) samples were positive in culture (four kidneys and four livers), corresponding to five animals with positive serology (one animal simultaneously positive for both kidney and liver) and two negatives. PCR detected Leptospira spp. in 14 samples (seven kidneys and seven livers) corresponding to 12 positive animals (two animals simultaneously positive for kidney and liver), of which ten were serologically positive and two negative. Conclusions PCR was faster, more practical and more sensitive than culture for detecting leptospires. The results reinforce the importance of sheep in the epidemiological context of leptospirosis. PMID:24822059

  11. Sample size and number of outcome measures of veterinary randomised controlled trials of pharmaceutical interventions funded by different sources, a cross-sectional study.

    PubMed

    Wareham, K J; Hyde, R M; Grindlay, D; Brennan, M L; Dean, R S

    2017-10-04

    Randomised controlled trials (RCTs) are a key component of the veterinary evidence base. Sample sizes and defined outcome measures are crucial components of RCTs. To describe the sample size and number of outcome measures of veterinary RCTs either funded by the pharmaceutical industry or not, published in 2011. A structured search of PubMed identified RCTs examining the efficacy of pharmaceutical interventions. Number of outcome measures, number of animals enrolled per trial, whether a primary outcome was identified, and the presence of a sample size calculation were extracted from the RCTs. The source of funding was identified for each trial and groups compared on the above parameters. Literature searches returned 972 papers; 86 papers comprising 126 individual trials were analysed. The median number of outcomes per trial was 5.0; there were no significant differences across funding groups (p = 0.133). The median number of animals enrolled per trial was 30.0; this was similar across funding groups (p = 0.302). A primary outcome was identified in 40.5% of trials and was significantly more likely to be stated in trials funded by a pharmaceutical company. A very low percentage of trials reported a sample size calculation (14.3%). Failure to report primary outcomes, justify sample sizes and the reporting of multiple outcome measures was a common feature in all of the clinical trials examined in this study. It is possible some of these factors may be affected by the source of funding of the studies, but the influence of funding needs to be explored with a larger number of trials. Some veterinary RCTs provide a weak evidence base and targeted strategies are required to improve the quality of veterinary RCTs to ensure there is reliable evidence on which to base clinical decisions.

  12. Electrochemical Immunosensor for the Detection of Aflatoxin B₁ in Palm Kernel Cake and Feed Samples.

    PubMed

    Azri, Farah Asilah; Selamat, Jinap; Sukor, Rashidah

    2017-11-30

    Palm kernel cake (PKC) is the solid residue following oil extraction of palm kernels and useful to fatten animals either as a single feed with only minerals and vitamins supplementation, or mixed with other feedstuffs such as corn kernels or soy beans. The occurrence of mycotoxins (aflatoxins, ochratoxins, zearalenone, and fumonisins) in feed samples affects the animal's health and also serves as a secondary contamination to humans via consumption of eggs, milk and meats. Of these, aflatoxin B₁ (AFB₁) is the most toxically potent and a confirmed carcinogen to both humans and animals. Methods such as High Performance Liquid Chromatography (HPLC) and Liquid Chromatography-Mass Spectrometry (LC-MS/MS) are common in the determination of mycotoxins. However, these methods usually require sample pre-treatment, extensive cleanup and skilled operator. Therefore, in the present work, a rapid method of electrochemical immunosensor for the detection of AFB₁ was developed based on an indirect competitive enzyme-linked immunosorbent assay (ELISA). Multi-walled carbon nanotubes (MWCNT) and chitosan (CS) were used as the electrode modifier for signal enhancement. N -ethyl- N '-(3-dimethylaminopropyl)-carbodiimide (EDC) and N -hydroxysuccinimide (NHS) activated the carboxyl groups at the surface of nanocomposite for the attachment of AFB₁-BSA antigen by covalent bonding. An indirect competitive reaction occurred between AFB₁-BSA and free AFB₁ for the binding site of a fixed amount of anti-AFB₁ antibody. A catalytic signal based on horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H₂O₂) and 3,3',5,5'-tetramethylbenzidine (TMB) mediator was observed as a result of attachment of the secondary antibody to the immunoassay system. As a result, the reduction peak of TMB (Ox) was measured by using differential pulse voltammetry (DPV) analysis. Based on the results, the electrochemical surface area was increased from 0.396 cm² to 1.298 cm² due to the electrode modification with MWCNT/CS. At the optimal conditions, the working range of the electrochemical immunosensor was from 0.0001 to 10 ng/mL with limit of detection of 0.1 pg/mL. Good recoveries were obtained for the detection of spiked feed samples (PKC, corn kernels, soy beans). The developed method could be used for the screening of AFB₁ in real samples.

  13. Evaluation of hazardous chemicals in edible insects and insect-based food intended for human consumption.

    PubMed

    Poma, Giulia; Cuykx, Matthias; Amato, Elvio; Calaprice, Chiara; Focant, Jean Francois; Covaci, Adrian

    2017-02-01

    Due to the rapid increase in world population, the waste of food and resources, and non-sustainable food production practices, the use of alternative food sources is currently strongly promoted. In this perspective, insects may represent a valuable alternative to main animal food sources due to their nutritional value and sustainable production. However, edible insects may be perceived as an unappealing food source and are indeed rarely consumed in developed countries. The food safety of edible insects can thus contribute to the process of acceptance of insects as an alternative food source, changing the perception of developed countries regarding entomophagy. In the present study, the levels of organic contaminants (i.e. flame retardants, PCBs, DDT, dioxin compounds, pesticides) and metals (As, Cd, Co, Cr, Cu, Ni, Pb, Sn, Zn) were investigated in composite samples of several species of edible insects (greater wax moth, migratory locust, mealworm beetle, buffalo worm) and four insect-based food items currently commercialized in Belgium. The organic chemical mass fractions were relatively low (PCBs: 27-2065 pg/g ww; OCPs: 46-368 pg/g ww; BFRs: up to 36 pg/g ww; PFRs 783-23800 pg/g ww; dioxin compounds: up to 0.25 pg WHO-TEQ/g ww) and were generally lower than those measured in common animal products. The untargeted screening analysis revealed the presence of vinyltoluene, tributylphosphate (present in 75% of the samples), and pirimiphos-methyl (identified in 50% of the samples). The levels of Cu and Zn in insects were similar to those measured in meat and fish in other studies, whereas As, Co, Cr, Pb, Sn levels were relatively low in all samples (<0.03 mg/kg ww). Our results support the possibility to consume these insect species with no additional hazards in comparison to the more commonly consumed animal products. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Use of artificial intelligence to identify cardiovascular compromise in a model of hemorrhagic shock.

    PubMed

    Glass, Todd F; Knapp, Jason; Amburn, Philip; Clay, Bruce A; Kabrisky, Matt; Rogers, Steven K; Garcia, Victor F

    2004-02-01

    To determine whether a prototype artificial intelligence system can identify volume of hemorrhage in a porcine model of controlled hemorrhagic shock. Prospective in vivo animal model of hemorrhagic shock. Research foundation animal surgical suite; computer laboratories of collaborating industry partner. Nineteen, juvenile, 25- to 35-kg, male and female swine. Anesthetized animals were instrumented for arterial and systemic venous pressure monitoring and blood sampling, and a splenectomy was performed. Following a 1-hr stabilization period, animals were hemorrhaged in aliquots to 10, 20, 30, 35, 40, 45, and 50% of total blood volume with a 10-min recovery between each aliquot. Data were downloaded directly from a commercial monitoring system into a proprietary PC-based software package for analysis. Arterial and venous blood gas values, glucose, and cardiac output were collected at specified intervals. Electrocardiogram, electroencephalogram, mixed venous oxygen saturation, temperature (core and blood), mean arterial pressure, pulmonary artery pressure, central venous pressure, pulse oximetry, and end-tidal CO(2) were continuously monitored and downloaded. Seventeen of 19 animals (89%) died as a direct result of hemorrhage. Stored data streams were analyzed by the prototype artificial intelligence system. For this project, the artificial intelligence system identified and compared three electrocardiographic features (R-R interval, QRS amplitude, and R-S interval) from each of nine unknown samples of the QRS complex. We found that the artificial intelligence system, trained on only three electrocardiographic features, identified hemorrhage volume with an average accuracy of 91% (95% confidence interval, 84-96%). These experiments demonstrate that an artificial intelligence system, based solely on the analysis of QRS amplitude, R-R interval, and R-S interval of an electrocardiogram, is able to accurately identify hemorrhage volume in a porcine model of lethal hemorrhagic shock. We suggest that this technology may represent a noninvasive means of assessing the physiologic state during and immediately following hemorrhage. Point of care application of this technology may improve outcomes with earlier diagnosis and better titration of therapy of shock.

  15. Analysis of Processed Foods Containing Oils and Fats by Time of Flight Mass Spectrometry with an APCI Direct Probe.

    PubMed

    Ito, Shihomi; Chikasou, Masato; Inohana, Shuichi; Fujita, Kazuhiro

    2016-01-01

    Discriminating vegetable oils and animal and milk fats by infrared absorption spectroscopy is difficult due to similarities in their spectral patterns. Therefore, a rapid and simple method for analyzing vegetable oils, animal fats, and milk fats using TOF/MS with an APCI direct probe ion source was developed. This method enabled discrimination of these oils and fats based on mass spectra and detailed analyses of the ions derived from sterols, even in samples consisting of only a few milligrams. Analyses of the mass spectra of processed foods containing oils and milk fats, such as butter, cheese, and chocolate, enabled confirmation of the raw material origin based on specific ions derived from the oils and fats used to produce the final product.

  16. Analysis of biofluids in aqueous environment based on mid-infrared spectroscopy.

    PubMed

    Fabian, Heinz; Lasch, Peter; Naumann, Dieter

    2005-01-01

    In this study we describe a semiautomatic Fourier transform infrared spectroscopic methodology for the analysis of liquid serum samples, which combines simple sample introduction with high sample throughput. The applicability of this new infrared technology to the analysis of liquid serum samples from a cohort of cattle naturally infected with bovine spongiform encephalopathy and from controls was explored in comparison to the conventional approach based on transmission infrared spectroscopy of dried serum films. Artifical neural network analysis of the infrared data was performed to differentiate between bovine spongiform encephalopathy-negative controls and animals in the late stage of the disease. After training of artifical neural network classifiers, infrared spectra of sera from an independent external validation data set were analyzed. In this way, sensitivities between 90 and 96% and specificities between 84 and 92% were achieved, respectively, depending upon the strategy of data collection and data analysis. Based on these results, the advantages and limitations of the liquid sample technique and the dried film approach for routine analysis of biofluids are discussed. 2005 Society of Photo-Optical Instrumentation Engineers.

  17. The Assessment of Animal Welfare in British Zoos by Government-Appointed Inspectors.

    PubMed

    Draper, Chris; Harris, Stephen

    2012-09-28

    We analysed the reports of government-appointed inspectors from 192 zoos between 2005-2008 to provide the first review of how animal welfare was assessed in British zoos since the enactment of the Zoo Licensing Act 1981. We examined the effects of whether or not a veterinarian was included in the inspection team, type of inspection, licence status of the zoo and membership of a zoo association on the inspectors' assessments of animal welfare standards in five areas that approximate to the Five Freedoms. At least 11% of full licence inspections did not comply with the legal requirement for two inspectors. The inspectors' reports were unclear as to how animal welfare was assessed, whether all animals or only a sub-sample had been inspected, and were based predominantly on welfare inputs rather than outcomes. Of 9,024 animal welfare assessments across the 192 zoos, 7,511 (83%) were graded as meeting the standards, 782 (9%) as substandard and the rest were not graded. Of the 192 zoos, 47 (24%) were assessed as meeting all the animal welfare standards. Membership of a zoo association was not associated with a higher overall assessment of animal welfare standards, and specialist collections such as Farm Parks and Other Bird collections performed least well. We recommend a number of changes to the inspection process that should lead to greater clarity in the assessment of animal welfare in British zoos.

  18. Molecular detection of Histoplasma capsulatum in insectivorous and frugivorous bats in Southeastern Brazil.

    PubMed

    Dos Santos, Bruna; Langoni, Helio; da Silva, Rodrigo Costa; Menozzi, Benedito Donizete; Bosco, Sandra de Moraes Gimenes; Paiz, Laís Moraes; Augusto, Livia Carla Ramos; Richini-Pereira, Virgínia Bodelão

    2017-12-27

    Bats are considered to play a significant role in the epidemiology of histoplasmosis, worldwide. We investigated the occurrence of H. capsulatum in lung samples from 89 bats, from urban areas in Southeastern Brazil, using nested PCR based on ribosomal DNA. Fungal DNA was detected in 31/89 samples (34.8%), of which 13/31 were Molossids (41.9%), 4/31 Eumops spp. (12.9%), 2/31 Artibeus lituratus (6.5%), and 12/31 others (38.7%). This is the first report of natural infection by H. capsulatum in A. lituratus in Southeastern Brazil, which reinforces the importance of these synanthropic animals in the epidemiology of histoplasmosis in urban areas. © The Author(s) 2017. 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.

  19. Effects of habitat disturbance on tropical forest biodiversity

    PubMed Central

    Alroy, John

    2017-01-01

    It is widely expected that habitat destruction in the tropics will cause a mass extinction in coming years, but the potential magnitude of the loss is unclear. Existing literature has focused on estimating global extinction rates indirectly or on quantifying effects only at local and regional scales. This paper directly predicts global losses in 11 groups of organisms that would ensue from disturbance of all remaining tropical forest habitats. The results are based on applying a highly accurate method of estimating species richness to 875 ecological samples. About 41% of the tree and animal species in this dataset are absent from disturbed habitats, even though most samples do still represent forests of some kind. The individual figures are 30% for trees and 8–65% for 10 animal groups. Local communities are more robust to disturbance because losses are partially balanced out by gains resulting from homogenization. PMID:28461482

  20. [Analysis on the results of etiology and serology of plague in Qinghai province from 2001 to 2010].

    PubMed

    Yang, Yonghai; Wang, Mei; Zhao, Xiaolong; Zhao, Zhongzhi; Zhang, Aiping; Wei, Rongjie; Wei, Baiqing; Wang, Zuyun

    2014-02-01

    To analyze the results of etiology and serology of plague among human and infected animals in Qinghai province from 2001 to 2010. Thirty-seven cases of human infected with plague, 53 541 different animal samples, 5 685 sets of vector insects flea and 49 039 different animal serum samples were obtained between 2001 and 2010. A total of 7 811 samples of serum from healthy farmers and herdsmen in 14 counties in Qinghai from 2005 to 2007 were collected. Yersinia pestis (Y. pestis) were detected in visceral and secretions from human, infected animals and vector insects, respectively. Plague antigen was detected by reverse indirect hemagglutination assay (RIHA) in those samples. Indirect hemagglutination assay (IHA) was used to test plague FI antibody in serum of human and infected animals. 37 human plague cases were confirmed, 21 strains of plague Y. pestis were isolated from human cases and 14 positive were detected out. 133 of 7 811 samples of human serum were IHA positive, with the positive rate at 1.7%. A total of 146 strains of plague were isolated from infected animals and vector insects, 99 out of which were from infected animals, with a ratio of Marmota himalayan at 72.7% (72/99) and the other 47 were from vector insects, with a ratio of callopsylla solaris at 68.1% (32/47). The number of IHA and PIHA positive were 300 and 10, respectively. A total of 3 animals and 3 insects species were identified as new epidemic hosts for plague. The natural plague focus of Microtus fuscus was discovered and confirmed and coexisted with natural focus of Marmota himalayan in Chengduo county, Yushu prefecture. The epidemic situation of plague is distributed mainly in Haixi, Yushu and Hainan prefectures. From 2001 to 2010, animal infected with plague was detected in successive years and human plague was very common in Qinghai. New infected animals and vector insects species and new epidemic areas were confirmed, hence the trend of plague prevalence for humans and animals is very active in Qinghai province.

  1. 9 CFR 145.11 - Supervision.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Supervision. 145.11 Section 145.11 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... sample collecting provided for in § 145.14 and may designate qualified persons as Authorized Testing...

  2. 9 CFR 145.11 - Supervision.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Supervision. 145.11 Section 145.11 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... sample collecting provided for in § 145.14 and may designate qualified persons as Authorized Testing...

  3. 9 CFR 145.11 - Supervision.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Supervision. 145.11 Section 145.11 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... sample collecting provided for in § 145.14 and may designate qualified persons as Authorized Testing...

  4. Molecular characterization of Hepatozoon sp. in cats from São Luís Island, Maranhão, Northeastern Brazil.

    PubMed

    de Bortoli, Caroline P; André, Marcos R; Braga, Maria do Socorro C; Machado, Rosangela Zacarias

    2011-10-01

    Few molecular studies have been done concerning the molecular characterization of Hepatozoon species among domestic and wild felids. The present work aimed to characterize molecularly the presence of Hepatozoon sp. DNA in cat blood samples from São Luís Island, Maranhão state, Northeastern Brazil. EDTA-whole blood samples were collected from 200 domestic cats with outdoor and wood areas access from São Luís, Maranhão, Brazil. Each sample of extracted DNA was used as a template in PCR reactions aiming to amplify a partial sequence of 18S rRNA of Hepatozoon spp. We also performed sequence alignment to establish the identity of the parasite species infecting these animals using DNA sequences based on 18S rRNA. From 200 sampled cats, Hepatozoon DNA was only found in one animal (0.5%). The found Hepatozoon DNA showed 97% of identity with Hemobartonella felis isolates 1 and 2 from Spain. When analyzing the phylogenetic tree, the found Hepatozoon DNA was in the same clade than H. felis isolates. Our findings suggest that more than one species of Hepatozoon could infect felids in Brazil.

  5. 9 CFR 91.3 - General export requirements.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    .... (c) Testing. All samples for tests required by §§ 91.5 through 91.13 for exportation of animals under... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false General export requirements. 91.3 Section 91.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF...

  6. 9 CFR 91.3 - General export requirements.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    .... (c) Testing. All samples for tests required by §§ 91.5 through 91.13 for exportation of animals under... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false General export requirements. 91.3 Section 91.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF...

  7. 9 CFR 91.3 - General export requirements.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    .... (c) Testing. All samples for tests required by §§ 91.5 through 91.13 for exportation of animals under... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false General export requirements. 91.3 Section 91.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF...

  8. 9 CFR 91.3 - General export requirements.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    .... (c) Testing. All samples for tests required by §§ 91.5 through 91.13 for exportation of animals under... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false General export requirements. 91.3 Section 91.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF...

  9. Detection and genotyping of group A rotaviruses isolated from sewage samples in Monastir, Tunisia between April 2007 and April 2010.

    PubMed

    Hassine-Zaafrane, M; Kaplon, J; Ben Salem, I; Sdiri-Loulizi, K; Sakly, N; Pothier, P; Aouni, M; Ambert-Balay, K

    2015-11-01

    To ascertain the viral load, the distribution of G and P types of group A rotaviruses (RV-A) in sewage samples and to compare strains in clinical, animal and environmental samples. During our study from April 2007 to April 2010, 518 samples of raw and treated sewage were collected from two biological sewage treatment plants (STPs) located in the Monastir region, Tunisia. RV-A was detected by real-time RT-PCR in 375 (72·4%) sewage samples. According to the quantification results of RV-A, it appears that the viral load in raw and treated sewage of the two STPs was quite similar (P = 0·735). The genotyping of RV-A strains detected in sewage samples showed a great diversity with 10 G types and 8 P types. Most of them were described as common in humans, but we also detected genotypes commonly found in animals. All the genotypes detected in two previous studies performed in our laboratory on clinical and bovine samples were also found in environmental samples. However, some genotypes commonly found in animal were only found in sewage samples. The comparison of environmental, clinical and animal data suggests that STPs may convey not only human sewage but also animal wastes, both of them contaminated with numerous RV-A strains which are not efficiently eliminated by the sewage treatment process and may spread to surface waters. This work demonstrates the potential release of human and animal RV-A into water sources, representing a public health risk, by inducing gastroenteritis in population, but also by increasing the risk of zoonotic transmission and formation of reassortant viruses which could get a higher infectious potential. Our findings also suggest that monitoring of sewage may provide an additional tool to determine the epidemiology of RV-A circulating in a given community. © 2015 The Society for Applied Microbiology.

  10. Sensitivity of bhk-21 cells supplemented with diethylaminoethyl-dextran for detection of street rabies virus in saliva samples.

    PubMed Central

    Larghi, O P; Nebel, A E; Lazaro, L; Savy, V L

    1975-01-01

    A tissue culture system for detecting rabies virus from saliva samples of suspected animals was developed and compared to suckling mouse inoculation. Swab samples were obtained from the mouth of the animal heads received for rabies diagnosis; these swabs were submerged in maintenance medium. The maintenance medium was inoculated intracerebrally into suckling mice and onto BHK-21 cells with diethylaminoethyl (DEAE)-dextran (BHK/DEAE) and without (BHK). Rabies immunofluorescence was performed on the brain of the mice dying during the observation period and also on both tissue culture systems every day after infection. The BHK-DEAE system detected 28 positive samples obtained from 48 rabid animals and the BHK system detected 18. By suckling mouse inoculation only 11 of the same positive samples were detected. A total of 90 samples was studied by the three methods. Rabies virus was detected by the tissue culture methods earlier than by suckling mouse inoculation. The BHK-DEAE method was an economic and fast method for rabies virus detection in saliva samples, which could be used for ecological and pathogenesis studies, as well for rabies diagnosis before the death of the suspected animal. PMID:1100655

  11. Evidence of presence of Mycobacterium tuberculosis in bovine tissue samples by multiplex PCR: possible relevance to reverse zoonosis.

    PubMed

    Mittal, M; Chakravarti, S; Sharma, V; Sanjeeth, B S; Churamani, C P; Kanwar, N S

    2014-04-01

    Bovine tuberculosis, caused by Mycobacterium bovis, remains one of the most important zoonotic health concerns worldwide. The transmission of Mycobacterium tuberculosis from humans to animals also occurs especially in countries where there is close interaction of humans with the animals. In the present study, thirty bovine lung tissue autopsy samples from an organized dairy farm located in North India were screened for the presence of Mycobacterium tuberculosis complex by smear microscopy, histopathological findings and PCR. Differential diagnosis of M. tuberculosis and M. bovis was made based on the deletion of mce-3 operon in M. bovis. The present study found eight of these samples positive for M. tuberculosis by multiplex PCR. Sequencing was performed on two PCR-positive representative samples and on annotation, and BLAST analysis confirmed the presence of gene fragment specific to Mycobacterium tuberculosis. The presence of M. tuberculosis in all the positive samples raises the possibility of human-to-cattle transmission and possible adaptation of this organism in bovine tissues. This study accentuates the importance of screening and differential diagnosis of Mycobacterium tuberculosis complex in humans and livestock for adopting effective TB control and eradication programmes. © 2014 Blackwell Verlag GmbH.

  12. Diversity of Cryptosporidium in brush-tailed rock-wallabies (Petrogale penicillata) managed within a species recovery programme

    PubMed Central

    Vermeulen, Elke T.; Ashworth, Deborah L.; Eldridge, Mark D.B.; Power, Michelle L.

    2015-01-01

    Host–parasite relationships are likely to be impacted by conservation management practices, potentially increasing the susceptibility of wildlife to emerging disease. Cryptosporidium, a parasitic protozoan genus comprising host-adapted and host-specific species, was used as an indicator of parasite movement between populations of a threatened marsupial, the brush-tailed rock-wallaby (Petrogale penicillata). PCR screening of faecal samples (n = 324) from seven wallaby populations across New South Wales, identified Cryptosporidium in 7.1% of samples. The sampled populations were characterised as captive, supplemented and wild populations. No significant difference was found in Cryptosporidium detection between each of the three population categories. The positive samples, detected using 18S rRNA screening, were amplified using the actin and gp60 loci. Multi-locus sequence analysis revealed the presence of Cryptosporidium fayeri, a marsupial-specific species, and C. meleagridis, which has a broad host range, in samples from the three population categories. Cryptosporidium meleagridis has not been previously reported in marsupials and hence the pathogenicity of this species to brush-tailed rock-wallabies is unknown. Based on these findings, we recommend further study into Cryptosporidium in animals undergoing conservation management, as well as surveying wild animals in release areas, to further understand the diversity and epidemiology of this parasite in threatened wildlife. PMID:25834789

  13. Refinement of a model of repeated cerebrospinal fluid collection in conscious rats.

    PubMed

    Amen, Eva Maria; Brecheisen, Muriel; Sach-Peltason, Lisa; Bergadano, Alessandra

    2017-02-01

    The cannulation of the cisterna magna in rats for in vivo sampling of cerebrospinal fluid serves as a valuable model for studying the delivery of new drugs into the central nervous system or disease models. It offers the advantages of repeated sampling without anesthesia-induced bias and using animals as their own controls. An established model was retrospectively reviewed for the outcomes and it was hypothesized that by refining the method, i.e. by (1) implementing pathophysiological-based anesthesia and analgesia, (2) using state-of-the-art peri-operative monitoring and supportive care, (3) increasing stability of the cement-cannula assembly, and (4) selecting a more adaptable animal strain, the outcome in using the model - quantified by peri-operative mortality, survival time and stability of the implant - could be improved and could enhance animal welfare. After refinement of the technique, peri-operative mortality decreased significantly (7 animals out of 73 compared with 4 out of 322; P = 0.001), survival time increased significantly (36 ± 14 days compared with 28 ± 18 days; P < 0.001), as well as the stability of the cement-cannula assembly (47 ± 8 days of adhesion compared with 33 ± 15 days and 34 ± 13 days using two other cement types; P < 0.001). Overall, the 3R concept of Russell and Burch was successfully addressed and animal welfare was improved by (1) the reduction in the total number of animals needed as a result of lower mortality or fewer euthanizations due to technical failure, and frequent use of individual rats over a time frame; and (2) improving the scientific quality of the model.

  14. Validation of the Filovirus Plaque Assay for Use in Preclinical Studies

    DTIC Science & Technology

    2016-09-02

    filoviruses in virus stocks, prepared viral challenge inocula and samples from research animals has recently been fully characterized and standardized for...and robust for filovirus titration in samples associated with the performance of GLP animal model studies. Keywords: Plaque assay; filovirus; Ebola...ebolavirus; marburgvirus; Marburg virus; Vero E6 cells; GLP compliant; validation; animal rule DISTRIBUTION STATEMENT A: Approved for public

  15. Evaluation of PMS-PCR technology for detection of Mycobacterium avium subsp. paratuberculosis directly from bovine fecal specimens.

    PubMed

    Salgado, M; Steuer, P; Troncoso, E; Collins, M T

    2013-12-27

    Mycobacterium avium subsp. paratuberculosis (MAP) causes paratuberculosis, or Johne's disease, in animals. Diagnosis of MAP infection is challenging because of the pathogen's fastidious in vitro growth requirements and low-level intermittent shedding in feces during the preclinical phase of the infection. Detection of these "low-shedders" is important for effective control of paratuberculosis as these animals serve as sources of infection for susceptible calves. Magnetic separation technology, used in combination with culture or molecular methods for the isolation and detection of pathogenic bacteria, enhances the analytical sensitivity and specificity of detection methods. The aim of the present study was to evaluate peptide-mediated magnetic separation (PMS) capture technology coupled with IS900 PCR using the Roche real-time PCR system (PMS-PCR), in comparison with fecal culture using BACTEC-MGIT 960 system, for detection of MAP in bovine fecal samples. Among the 351 fecal samples 74.9% (263/351) were PMS-PCR positive while only 12.3% (43/351) were MGIT culture-positive (p=0.0001). All 43 MGIT culture-positive samples were also positive by PMS-PCR. Mean PMS-PCR crossing-point (Cp) values for the 13 fecal samples with the highest number of MAP, based on time to detection, (26.3) were significantly lower than for the 17 fecal samples with <100 MAP per 2g feces (30.06) (p<0.05). PMS-PCR technology provided results in a shorter time and yielded a higher number of positive results than MGIT culture. Earlier and faster detection of animals shedding MAP by PMS-PCR should significantly strengthen control efforts for MAP-infected cattle herds by helping to limit infection transmission at earlier stages of the infection. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. qFibrosis: A fully-quantitative innovative method incorporating histological features to facilitate accurate fibrosis scoring in animal model and chronic hepatitis B patients

    PubMed Central

    Tai, Dean C.S.; Wang, Shi; Cheng, Chee Leong; Peng, Qiwen; Yan, Jie; Chen, Yongpeng; Sun, Jian; Liang, Xieer; Zhu, Youfu; Rajapakse, Jagath C.; Welsch, Roy E.; So, Peter T.C.; Wee, Aileen; Hou, Jinlin; Yu, Hanry

    2014-01-01

    Background & Aims There is increasing need for accurate assessment of liver fibrosis/cirrhosis. We aimed to develop qFibrosis, a fully-automated assessment method combining quantification of histopathological architectural features, to address unmet needs in core biopsy evaluation of fibrosis in chronic hepatitis B (CHB) patients. Methods qFibrosis was established as a combined index based on 87 parameters of architectural features. Images acquired from 25 Thioacetamide-treated rat samples and 162 CHB core biopsies were used to train and test qFibrosis and to demonstrate its reproducibility. qFibrosis scoring was analyzed employing Metavir and Ishak fibrosis staging as standard references, and collagen proportionate area (CPA) measurement for comparison. Results qFibrosis faithfully and reliably recapitulates Metavir fibrosis scores, as it can identify differences between all stages in both animal samples (p <0.001) and human biopsies (p <0.05). It is robust to sampling size, allowing for discrimination of different stages in samples of different sizes (area under the curve (AUC): 0.93–0.99 for animal samples: 1–16 mm2; AUC: 0.84–0.97 for biopsies: 10–44 mm in length). qFibrosis can significantly predict staging underestimation in suboptimal biopsies (<15 mm) and under- and over-scoring by different pathologists (p <0.001). qFibrosis can also differentiate between Ishak stages 5 and 6 (AUC: 0.73, p = 0.008), suggesting the possibility of monitoring intra-stage cirrhosis changes. Best of all, qFibrosis demonstrates superior performance to CPA on all counts. Conclusions qFibrosis can improve fibrosis scoring accuracy and throughput, thus allowing for reproducible and reliable analysis of efficacies of anti-fibrotic therapies in clinical research and practice. PMID:24583249

  17. Risk-based methods for fish and terrestrial animal disease surveillance.

    PubMed

    Oidtmann, Birgit; Peeler, Edmund; Lyngstad, Trude; Brun, Edgar; Bang Jensen, Britt; Stärk, Katharina D C

    2013-10-01

    Over recent years there have been considerable methodological developments in the field of animal disease surveillance. The principles of risk analysis were conceptually applied to surveillance in order to further develop approaches and tools (scenario tree modelling) to design risk-based surveillance (RBS) programmes. In the terrestrial animal context, examples of risk-based surveillance have demonstrated the substantial potential for cost saving, and a similar benefit is expected also for aquatic animals. RBS approaches are currently largely absent for aquatic animal diseases. A major constraint in developing RBS designs in the aquatic context is the lack of published data to assist in the design of RBS: this applies to data on (i) the relative risk of farm sites becoming infected due to the presence or absence of a given risk factor; (ii) the sensitivity of diagnostic tests (specificity is often addressed by follow-up investigation and re-testing and therefore less of a concern); (iii) data on the variability of prevalence of infection for fish within a holding unit, between holding units and at farm level. Another constraint is that some of the most basic data for planning surveillance are missing, e.g. data on farm location and animal movements. In Europe, registration or authorisation of fish farms has only recently become a requirement under EU Directive 2006/88. Additionally, the definition of the epidemiological unit (at site or area level) in the context of aquaculture is a challenge due to the often high level of connectedness (mainly via water) of aquaculture facilities with the aquatic environment. This paper provides a review of the principles, methods and examples of RBS in terrestrial, farmed and wild animals. It discusses the special challenges associated with surveillance for aquatic animal diseases (e.g. accessibility of animals for inspection and sampling, complexity of rearing systems) and provides an overview of current developments relevant for the design of RBS for fish diseases. Suggestions are provided on how the current constraints to applying RBS to fish diseases can be overcome. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  18. A meaty matter. Pet diet and the vegetarian's dilemma.

    PubMed

    Rothgerber, Hank

    2013-09-01

    The present research examined pet ownership, current pet diet, and guilt associated with pet diet among a fairly large sample of non-meat-eaters (n=515). It specifically focused on the conflict that pits feeding one's pet an animal-based diet that may be perceived as best promoting their well-being with concerns over animal welfare and environmental degradation threatened by such diets, here labeled the vegetarian's dilemma. Questionnaire responses indicated that ethically motivated meat abstainers were more likely to own pets and owned more of them than those motivated by health concerns or a combination of ethical and health concerns. Vegans and those resisting meat on ethical grounds were more likely to feed their pet a vegetarian diet and expressed the greatest concerns over feeding their pet an animal-based diet. For vegans and ethical meat abstainers, it is suggested that questions concerning what to feed their pet approaches a tragic tradeoff contrasting two sacred values: protecting the well-being of their pets and protecting the well-being of other animals and the environment. For meat abstainers motivated by health concerns, this constitutes a relatively easy moral problem because the primary concern for such individuals is the health of their pet with less or no regard for other ramifications of the decision, i.e., harming other animals or the environment. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Water quality in the upper Shoal Creek basin, southwestern Missouri, 1999-2000

    USGS Publications Warehouse

    Schumacher, John G.

    2001-01-01

    Results of a water-quality investigation of the upper Shoal Creek Basin in southwestern Missouri indicate that concentrations of total nitrite plus nitrate as nitrogen (NO2t+NO3t) in water samples from Shoal Creek were unusually large [mean of 2.90 mg/L (milligrams per liter), n (sample size)=60] compared to other Missouri streams (mean of 1.02 mg/L, n=1,340). A comparison of instantaneous base-flow loads of NO2t+NO3t indicates that at base-flow conditions, most NO2t+NO3t discharged by Shoal Creek is from nonpoint sources. Nearly all the base-flow instantaneous load of total phosphorus as P (Pt) discharged by Shoal Creek can be attributed to effluent from a municipal wastewater treatment plant. Samples collected from a single runoff event indicate that substantial quantities of Pt can be transported during runoff events compared to base-flow transport. Only minor quantities of NO2t+NO3t are transported during runoff events compared to base-flow transport. Fecal coliform bacteria densities at several locations exceed the Missouri Department of Natural Resources (MDNR) standard of 200 col/100 mL (colonies per 100 milliliters) for whole-body contact recreation. During 13 months of monitoring at 13 stream sites, fecal coliform densities (median of 277 and 400 col/100 mL) at two sites (sites 2 and 3) on Shoal Creek exceeded the MDNR standard at base-flow conditions. The maximum fecal coliform density of 120,000 col/100 mL was detected at site 3 (MDNR monitoring site) during a runoff event in April 1999 at a peak discharge of 1,150 ft3/s (cubic feet per second). Fecal coliform densities also exceeded the MDNR standard in three tributaries with the largest densities (median of 580 col/100 mL) detected in Pogue Creek. Results of ribopattern analyses indicate that most Escherichia coli (E. coli) bacteria in water samples from the study area probably are from nonhuman sources. The study area contains about 25,000 cattle, and has an estimated annual production of 33 million broilers and 300,000 turkeys. Probable nonhuman sources included turkeys, horses, chickens, and cattle; however, wildlife sources such as deer, raccoon, muskrat, and opossum were not evaluated. Human waste was an important source of E. coli in water samples collected at the MDNR monitoring site (site 3) on Shoal Creek and at two tributary sites (Joyce Creek and Woodward Creek). In general, the detection of human ribopatterns was consistent with the detection of organic compounds commonly associated with human wastewater such as caffeine, triclosan, or phenol, and the fecal indicators cholesterol and 3B-coprostanol. Ribopattern analysis indicate that horses were an important source of E. coli in Woodward Creek, which was consistent with horses being pastured immediately upstream from the sampling site on this creek. Pogue Creek contains a large density of turkey barns and five of eight E. coli isolates from one sample from Pogue Creek were matched to turkeys. Water samples from Pogue Creek generally did not contain detectable concentrations of human wastewater compounds, but one sample did contain detectable quantities of the antibiotics tylosin and lincomycin (widely used in the animal industry), and sulfamethoxazole (human use only). Although promising, the ability of ribopattern analyses to positively identify the source of a particular isolate is uncertain because of the small sample size, possible differences between animal source patterns in the study area and database used, lack of native wildlife source patterns, and variation in results depending on the number of possible animal host considered. Results of this study indicate that a trend of increasing fecal coliform densities with increasing time detected by the MDNR is, in part, caused by trends in annual precipitation and stream discharge, and not necessarily changes in land use or densities of animal operations. A multiple linear regression (MLR) model using specific conductance and wate

  20. Differential metabolite levels in response to spawning-induced inappetence in Atlantic salmon Salmo salar.

    PubMed

    Cipriano, Rocco C; Smith, McKenzie L; Vermeersch, Kathleen A; Dove, Alistair D M; Styczynski, Mark P

    2015-03-01

    Atlantic salmon Salmo salar undergo months-long inappetence during spawning, but it is not known whether this inappetence is a pathological state or one for which the fish are adapted. Recent work has shown that inappetent whale sharks can exhibit circulating metabolite profiles similar to ketosis known to occur in humans during starvation. In this work, metabolite profiling was used to explore differences in analyte profiles between a cohort of inappetent spawning run Atlantic salmon and captively reared animals that were fed up to and through the time of sampling. The two classes of animals were easily distinguished by their metabolite profiles. The sea-run fish had elevated ɷ-9 fatty acids relative to the domestic feeding animals, while other fatty acid concentrations were reduced. Sugar alcohols were generally elevated in inappetent animals, suggesting potentially novel metabolic responses or pathways in fish that feature these compounds. Compounds expected to indicate a pathological catabolic state were not more abundant in the sea-run fish, suggesting that the animals, while inappetent, were not stressed in an unnatural way. These findings demonstrate the power of discovery-based metabolomics for exploring biochemistry in poorly understood animal models. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Using Dried Blood Spot Sampling to Improve Data Quality and Reduce Animal Use in Mouse Pharmacokinetic Studies

    PubMed Central

    Wickremsinhe, Enaksha R; Perkins, Everett J

    2015-01-01

    Traditional pharmacokinetic analysis in nonclinical studies is based on the concentration of a test compound in plasma and requires approximately 100 to 200 µL blood collected per time point. However, the total blood volume of mice limits the number of samples that can be collected from an individual animal—often to a single collection per mouse—thus necessitating dosing multiple mice to generate a pharmacokinetic profile in a sparse-sampling design. Compared with traditional methods, dried blood spot (DBS) analysis requires smaller volumes of blood (15 to 20 µL), thus supporting serial blood sampling and the generation of a complete pharmacokinetic profile from a single mouse. Here we compare plasma-derived data with DBS-derived data, explain how to adopt DBS sampling to support discovery mouse studies, and describe how to generate pharmacokinetic and pharmacodynamic data from a single mouse. Executing novel study designs that use DBS enhances the ability to identify and streamline better drug candidates during drug discovery. Implementing DBS sampling can reduce the number of mice needed in a drug discovery program. In addition, the simplicity of DBS sampling and the smaller numbers of mice needed translate to decreased study costs. Overall, DBS sampling is consistent with 3Rs principles by achieving reductions in the number of animals used, decreased restraint-associated stress, improved data quality, direct comparison of interanimal variability, and the generation of multiple endpoints from a single study. PMID:25836959

  2. Pyrosequencing-based assessment of microbial community shifts in leachate from animal carcass burial lysimeter.

    PubMed

    Kim, Hyun Young; Seo, Jiyoung; Kim, Tae-Hun; Shim, Bomi; Cha, Seok Mun; Yu, Seungho

    2017-06-01

    This study examined the use of microbial community structure as a bio-indicator of decomposition levels. High-throughput pyrosequencing technology was used to assess the shift in microbial community of leachate from animal carcass lysimeter. The leachate samples were collected monthly for one year and a total of 164,639 pyrosequencing reads were obtained and used in the taxonomic classification and operational taxonomy units (OTUs) distribution analysis based on sequence similarity. Our results show considerable changes in the phylum-level bacterial composition, suggesting that the microbial community is a sensitive parameter affected by the burial environment. The phylum classification results showed that Proteobacteria (Pseudomonas) were the most influential taxa in earlier decomposition stage whereas Firmicutes (Clostridium, Sporanaerobacter, and Peptostreptococcus) were dominant in later stage under anaerobic conditions. The result of this study can provide useful information on a time series of leachate profiles of microbial community structures and suggest patterns of microbial diversity in livestock burial sites. In addition, this result can be applicable to predict the decomposition stages under clay loam based soil conditions of animal livestock. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Move Over! Studying Flatfish Travel Patterns to Profile Fish Behavior

    ERIC Educational Resources Information Center

    Aultman, Terry; Curran, Mary Carla

    2012-01-01

    The way an animal moves from place to place can inform us about its life and environment. In this lesson, students examine the travel patterns of juvenile flatfishes in an estuary. The process of sampling bottom-dwelling fishes is explained, and data from a university-based marine science laboratory are evaluated. Students compare the distance…

  4. Odor and odorous chemical emissions from animal buildings: Part 5 - Correlations between odor intensities and chemical concentrations (GC-MS/O)

    USDA-ARS?s Scientific Manuscript database

    Simultaneous chemical and sensory analysis based on gas chromatography-mass spectrometry-olfactometry (GC-MS-O) of air samples from livestock operations is a very useful approach for quantification of target odorous gases and also for ranking of odorous compounds. This information can help link spec...

  5. Fabrication and characterization of polymer blends and composites derived from biopolymers

    NASA Astrophysics Data System (ADS)

    Sharma, Suraj

    This research focuses on fabricating blends and composites from natural polymers especially from proteins and natural epoxy, and describing the properties of plastics made from them. Specifically, plastic samples from partially denatured feathermeal and bloodmeal proteins, derived from the animal co-products (rendering) industry, were successfully produced through a compression molding process. The modulus (stiffness) of the material obtained was found to be comparable with that of commercial synthetic materials, such as polystyrene, but was found to have lower toughness characteristics, which is a common phenomenon among plastics produced from animal and plant proteins. Therefore, this study explored blending methods for improving the toughness. Plastic forming conditions for undenatured animal proteins such as chicken egg whites albumin and whey, used as a model, were established to prepare plastics from their blends with animal co-product proteins. The resultant plastic samples from these biomacromolecular blends demonstrated improved mechanical properties that were also compared with the established theoretical models known for polymer blends and composites. Moreover, plastics from albumin of chicken egg whites and human serum have demonstrated their potential in medical applications that require antibacterial properties. Another natural polymer vegetable oil-based epoxy, especially epoxidized linseed oil, showed significant potential to replace petroleum-derived resins for use as a matrix for composites in structural applications. Moreover, the research showed the benefits of ultrasonic curing, which can help in preparing the out-of-autoclave composites.

  6. 2,3,7,8-DIBENZO-P-DIOXINS IN MINED CLAY PRODUCTS ...

    EPA Pesticide Factsheets

    Ball clay was the source of dioxin contamination discovered in selected chickens analyzed as part of a joint U.S. Department of Agriculture/U.S. Environmental Protection Agency national survey of the U.S. poultry supply conducted in 1997. The affected animals, which had been raised in the southern United States, represented approximately 5% of the national poultry production . All of these chickens and other animal food sources (i.e., farm-raised catfish), similarly contaminated, were fed a diet of animal feed containing ball clay as an anti-caking additive. The clay was mined in northwestern Mississippi within a geological formation referred to as the Mississippi Embayment. Individual raw and processed ball clay samples were analyzed for the presence of the 2,3,7,8-PCDDs/PCDFs. The average toxic equivalents (TEQs) for the raw and processed samples were 1513 and 996 ppt dry weight, respectively. Other mined clay-based products used in animal feeds revealed lower TEQs. All of the products exhibited either an absence of detectable concentrations of 2,3,7,8-PCDFs or concentrations 2-3 orders of magnitude lower than the PCDDs. The isomer distribution, specific isomer identification, and congener profile of the PCDDs in the clay were established and compared to known sources of dioxin contamination. Several unique features of this isomer distribution are characteristic of the clays and are distinguishable from those other known sources. These characteristic

  7. Quantifying individual variation in the propensity to attribute incentive salience to reward cues.

    PubMed

    Meyer, Paul J; Lovic, Vedran; Saunders, Benjamin T; Yager, Lindsay M; Flagel, Shelly B; Morrow, Jonathan D; Robinson, Terry E

    2012-01-01

    If reward-associated cues acquire the properties of incentive stimuli they can come to powerfully control behavior, and potentially promote maladaptive behavior. Pavlovian incentive stimuli are defined as stimuli that have three fundamental properties: they are attractive, they are themselves desired, and they can spur instrumental actions. We have found, however, that there is considerable individual variation in the extent to which animals attribute Pavlovian incentive motivational properties ("incentive salience") to reward cues. The purpose of this paper was to develop criteria for identifying and classifying individuals based on their propensity to attribute incentive salience to reward cues. To do this, we conducted a meta-analysis of a large sample of rats (N = 1,878) subjected to a classic Pavlovian conditioning procedure. We then used the propensity of animals to approach a cue predictive of reward (one index of the extent to which the cue was attributed with incentive salience), to characterize two behavioral phenotypes in this population: animals that approached the cue ("sign-trackers") vs. others that approached the location of reward delivery ("goal-trackers"). This variation in Pavlovian approach behavior predicted other behavioral indices of the propensity to attribute incentive salience to reward cues. Thus, the procedures reported here should be useful for making comparisons across studies and for assessing individual variation in incentive salience attribution in small samples of the population, or even for classifying single animals.

  8. Adult-Perpetrated Animal Abuse: A Systematic Literature Review.

    PubMed

    Alleyne, Emma; Parfitt, Charlotte

    2017-01-01

    Adults perpetrate the majority of animal abuse incidents yet clinicians are left with very little evidence base to advance/enhance their practice. The purpose of this systematic review is to synthesize and evaluate the current literature on adult-perpetrated animal abuse and to identify the etiological factors related to this type of offending. Twenty-three studies met the specific inclusion criteria but most importantly, they examined the characteristics of adult perpetrators of animal abuse. The findings from this review were demarcated by sample type: (1) Participants were the perpetrators of the animal abuse or held offense-supportive attitudes and (2) participants were victims of intimate partner violence reporting incidents of animal abuse perpetrated by their partner. From the perpetrator perspective, there were key developmental (i.e., maladaptive parenting strategies), behavioral (such as varied offending behaviors), and psychological (e.g., callousness, empathy deficits) factors highlighted in the literature. Finally, in the context of intimate partner violence, findings indicated that perpetrators abuse animals to control, coerce, intimidate, and/or manipulate their victims (this effect is moderated by the victims' emotional attachment to their pet). This review inherently underlines treatment targets that could achieve greater clinical gains, but we also conclude that more empirical and theoretical work is needed in order to set an agenda that prioritizes future research and effective practice.

  9. High-Resolution Laser-Induced Breakdown Spectroscopy used in Homeland Security and Forensic Applications

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

    Martin, Madhavi Z; Wullschleger, Stan D; Vass, Arpad Alexander

    The technique of laser-induced breakdown spectroscopy (LIBS) to detect elements for a variety of homeland security applications such as nuclear materials identification and inventory,and forensic applications has been demonstrated. For nuclear materials applications, we detected and profiled metals in coatings that were used to encapsulate nuclear fuel. Multivariate analysis has been successfully employed in the quantification of elements present in treated wood and engineered wood composites. These examples demonstrate that LIBS-based techniques are inherently well suited for diverse environmental applications related to homeland security. Three key advantages are evident: (1) small samples (mg) are sufficient; (2) samples can be analyzedmore » by LIBS very rapidly, and (3) biological materials such as human and animal bones and wood can be analyzed with minimal sample preparation. For forensic applications they have used LIBS to determine differences in animal and human bones. They have also applied this technique in the determination of counterfeit and non-counterfeit currency. They recently applied LIBS in helping to solve a murder case.« less

  10. Dyspepsia treatment with Al compounds widely used in clinical practice — an animal model approach

    NASA Astrophysics Data System (ADS)

    Pinheiro, T.; Canena, J.; Reis, J.; Santos, A. M.; Pinto, A. S.; Quina, M. G.; Reis, M. A.; Alves, L. C.

    1996-04-01

    The potential toxic effects of Al to organs and tissues used in drugs commonly applied in dyspepsia as therapeutic, have been studied. Brain, liver, kidney and serum samples obtained from Wistar rats treated with two commercial Al complexes were studied and compared with equivalent samples collected from healthy animals receiving a placebo. The major alterations found, connected with the persistent intake of medicaments based on Al compounds, are relative to the accumulation of Al in liver and kidney. Also, the Al levels increase in brain and serum of rats supplemented with one of the Al complexes used. In the liver and kidney samples analyzed alterations in the Cu and Zn content levels were observed. Furthermore, a tendency to the decrease of Fe content in kidney and an increase of the Mn levels in brain is observed. The elemental alterations found are probably related to the intake of the drugs tested and are dependent on the type of the Al complex administered. The results obtained suggest that the long term use of these medicaments in the clinical practice should be thought over.

  11. Possible sources of nitrate in ground water at swine licensed-managed feeding operations in Oklahoma, 2001

    USGS Publications Warehouse

    Becker, Mark F.; Peter, Kathy D.; Masoner, Jason

    2002-01-01

    Samples collected and analyzed by the Oklahoma Department of Agriculture, Food, and Forestry from 1999 to 2001 determined that nitrate exceeded the U.S. Environmental Protection Agency maximum contaminant level for public drinking-water supplies of 10 milligrams per liter as nitrogen in 79 monitoring wells at 35 swine licensed-managed feeding operations (LMFO) in Oklahoma. The LMFOs are located in rural agricultural settings where long-term agriculture has potentially affected the ground-water quality in some areas. Land use prior to the construction of the LMFOs was assessed to evaluate the types of agricultural land use within a 500-meter radius of the sampled wells. Chemical and microbiological techniques were used to determine the possible sources of nitrate in water sampled from 10 wastewater lagoons and 79 wells. Samples were analyzed for dissolved major ions, dissolved trace elements, dissolved nutrients, nitrogen isotope ratios of nitrate and ammonia, wastewater organic compounds, and fecal coliform bacteria. Bacteria ribotyping analysis was done on selected samples to identify possible specific animal sources. A decision process was developed to identify the possible sources of nitrate. First, nitrogen isotope ratios were used to define sources as animal, mixed animal and fertilizer, or fertilizer. Second, wastewater organic compound detections, nitrogen-isotope ratios, fecal coliform bacteria detections, and ribotyping were used to refine the identification of possible sources as LFMO waste, fertilizer, or unidentified animal or mixtures of these sources. Additional evidence provided by ribotyping and wastewater organic compound data can, in some cases, specifically indicate the animal source. Detections of three or more wastewater organic compounds that are indicators of animal sources and detections of fecal coliform bacteria provided additional evidence of an animal source. LMFO waste was designated as a possible source of nitrate in water from 10 wells. The source of waste in water from five of those wells was determined through ribotyping, and the source of waste in water from the remaining five wells was determined by detections of three or more animal-waste compounds in the well samples. LMFO waste in the water from wells with unidentified animal source of nitrate does not indicate that LMFO waste was not the source, but indicated that multiple animal sources, including LMFO waste, may be the source of the nitrate.

  12. From the field: Efficacy of detecting Chronic Wasting Disease via sampling hunter-killed white-tailed deer

    USGS Publications Warehouse

    Diefenbach, D.R.; Rosenberry, C.S.; Boyd, Robert C.

    2004-01-01

    Surveillance programs for Chronic Wasting Disease (CWD) in free-ranging cervids often use a standard of being able to detect 1% prevalence when determining minimum sample sizes. However, 1% prevalence may represent >10,000 infected animals in a population of 1 million, and most wildlife managers would prefer to detect the presence of CWD when far fewer infected animals exist. We wanted to detect the presence of CWD in white-tailed deer (Odocoileus virginianus) in Pennsylvania when the disease was present in only 1 of 21 wildlife management units (WMUs) statewide. We used computer simulation to estimate the probability of detecting CWD based on a sampling design to detect the presence of CWD at 0.1% and 1.0% prevalence (23-76 and 225-762 infected deer, respectively) using tissue samples collected from hunter-killed deer. The probability of detection at 0.1% prevalence was <30% with sample sizes of ???6,000 deer, and the probability of detection at 1.0% prevalence was 46-72% with statewide sample sizes of 2,000-6,000 deer. We believe that testing of hunter-killed deer is an essential part of any surveillance program for CWD, but our results demonstrated the importance of a multifaceted surveillance approach for CWD detection rather than sole reliance on testing hunter-killed deer.

  13. Surveillance of Salmonella prevalence in animal feeds and characterization of the Salmonella isolates by serotyping and antimicrobial susceptibility.

    PubMed

    Li, X; Bethune, L A; Jia, Y; Lovell, R A; Proescholdt, T A; Benz, S A; Schell, T C; Kaplan, G; McChesney, D G

    2012-08-01

    This article presents the surveillance data from the Feed Contaminants Program (2002-2009) and Salmonella Assignment (2007-2009) of the U.S. Food and Drug Administration (FDA), which monitor the trend of Salmonella contamination in animal feeds. A total of 2,058 samples were collected from complete animal feeds, feed ingredients, pet foods, pet treats, and supplements for pets in 2002-2009. These samples were tested for the presence of Salmonella. Those that were positive for Salmonella underwent serotyping and testing for antimicrobial susceptibility. Of the 2,058 samples, 257 were positive for Salmonella (12.5%). The results indicate a significant overall Salmonella reduction (p≤0.05) in animal feeds from 18.2% (187 samples tested) in 2002 to 8.0% (584 samples tested) in 2009. Among these samples, feed ingredients and pet foods/treats had the most significant reduction (p≤0.05). Of the 45 Salmonella serotypes identified, Salmonella Senftenberg and Salmonella Montevideo were the top two common serotypes (8.9%). Of the 257 Salmonella isolates obtained, 54 isolates (21%) were resistant to at least one antimicrobial. The findings provide the animal feed industries with Salmonella prevalence information that can be used to address Salmonella contamination problems. Our findings can also be used to educate pet owners when handling pet foods and treats at home to prevent salmonellosis.

  14. 9 CFR 113.105 - Leptospira Hardjo Bacterin.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Leptospira Hardjo Bacterin. 113.105 Section 113.105 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF... unsatisfactory by any prescribed test shall not be released. (a) Purity test. Final container samples of...

  15. Occurrence and prevalence of Cronobacter spp. in plant and animal derived food sources: a systematic review and meta-analysis.

    PubMed

    Sani, Norrakiah Abdullah; Odeyemi, Olumide A

    2015-01-01

    Cronobacter species are motile, non-spore forming, Gram negative emerging opportunistic pathogens mostly associated with bacteremia, meningitis, septicemia, brain abscesses and necrotizing enterocolitis in infected neonates, infants and immunocompromised adults. Members of the genus Cronobacter are previously associated with powdered infant formula although the main reservoir and routes of contamination are yet to be ascertained. This study therefore aim to summarize occurrence and prevalence of Cronobacter spp. from different food related sources. A retrospective systematic review and meta-analysis of peer reviewed primary studies reported between 2008 and 2014 for the occurrence and prevalence of Cronobacter spp. in animal and plant related sources was conducted using "Cronobacter isolation", "Cronobacter detection" and "Cronobacter enumeration" as search terms in the following databases: Web of Science (Science Direct) and ProQuest. Data extracted from the primary studies were then analyzed with meta-analysis techniques for effect rate and fixed effects was used to explore heterogeneity between the sources. Publication bias was evaluated using funnel plot. A total of 916 articles were retrieved from the data bases of which 28 articles met inclusion criteria. Cronobacter spp. could only be isolated from 103 (5.7 %) samples of animal related food while 123 (19 %) samples of plant related food samples harbors the bacteria. The result of this study shows that occurrence of Cronobacter was more prevalent in plant related sources with overall prevalence rate of 20.1 % (95 % CI 0.168-0.238) than animal originated sources with overall prevalence rate of 8 % (95 % CI 0.066-0.096). High heterogeneity (I (2) = 84) was observed mostly in plant related sources such as herbs, spices and vegetables compared to animal related sources (I (2) = 82). It could be observed from this study that plant related sources serve as reservoir and contamination routes of Cronobacter spp.

  16. Comparing Microbiome Sampling Methods in a Wild Mammal: Fecal and Intestinal Samples Record Different Signals of Host Ecology, Evolution

    PubMed Central

    Ingala, Melissa R.; Simmons, Nancy B.; Wultsch, Claudia; Krampis, Konstantinos; Speer, Kelly A.; Perkins, Susan L.

    2018-01-01

    The gut microbiome is a community of host-associated symbiotic microbes that fulfills multiple key roles in host metabolism, immune function, and tissue development. Given the ability of the microbiome to impact host fitness, there is increasing interest in studying the microbiome of wild animals to better understand these communities in the context of host ecology and evolution. Human microbiome research protocols are well established, but wildlife microbiome research is still a developing field. Currently, there is no standardized set of best practices guiding the collection of microbiome samples from wildlife. Gut microflora are typically sampled either by fecal collection, rectal swabbing, or by destructively sampling the intestinal contents of the host animal. Studies rarely include more than one sampling technique and no comparison of these methods currently exists for a wild mammal. Although some studies have hypothesized that the fecal microbiome is a nested subset of the intestinal microbiome, this hypothesis has not been formally tested. To address these issues, we examined guano (feces) and distal intestinal mucosa from 19 species of free-ranging bats from Lamanai, Belize, using 16S rRNA amplicon sequencing to compare microbial communities across sample types. We found that the diversity and composition of intestine and guano samples differed substantially. In addition, we conclude that signatures of host evolution are retained by studying gut microbiomes based on mucosal tissue samples, but not fecal samples. Conversely, fecal samples retained more signal of host diet than intestinal samples. These results suggest that fecal and intestinal sampling methods are not interchangeable, and that these two microbiotas record different information about the host from which they are isolated. PMID:29765359

  17. Comparing Microbiome Sampling Methods in a Wild Mammal: Fecal and Intestinal Samples Record Different Signals of Host Ecology, Evolution.

    PubMed

    Ingala, Melissa R; Simmons, Nancy B; Wultsch, Claudia; Krampis, Konstantinos; Speer, Kelly A; Perkins, Susan L

    2018-01-01

    The gut microbiome is a community of host-associated symbiotic microbes that fulfills multiple key roles in host metabolism, immune function, and tissue development. Given the ability of the microbiome to impact host fitness, there is increasing interest in studying the microbiome of wild animals to better understand these communities in the context of host ecology and evolution. Human microbiome research protocols are well established, but wildlife microbiome research is still a developing field. Currently, there is no standardized set of best practices guiding the collection of microbiome samples from wildlife. Gut microflora are typically sampled either by fecal collection, rectal swabbing, or by destructively sampling the intestinal contents of the host animal. Studies rarely include more than one sampling technique and no comparison of these methods currently exists for a wild mammal. Although some studies have hypothesized that the fecal microbiome is a nested subset of the intestinal microbiome, this hypothesis has not been formally tested. To address these issues, we examined guano (feces) and distal intestinal mucosa from 19 species of free-ranging bats from Lamanai, Belize, using 16S rRNA amplicon sequencing to compare microbial communities across sample types. We found that the diversity and composition of intestine and guano samples differed substantially. In addition, we conclude that signatures of host evolution are retained by studying gut microbiomes based on mucosal tissue samples, but not fecal samples. Conversely, fecal samples retained more signal of host diet than intestinal samples. These results suggest that fecal and intestinal sampling methods are not interchangeable, and that these two microbiotas record different information about the host from which they are isolated.

  18. Accounting for Location Error in Kalman Filters: Integrating Animal Borne Sensor Data into Assimilation Schemes

    PubMed Central

    Sengupta, Aritra; Foster, Scott D.; Patterson, Toby A.; Bravington, Mark

    2012-01-01

    Data assimilation is a crucial aspect of modern oceanography. It allows the future forecasting and backward smoothing of ocean state from the noisy observations. Statistical methods are employed to perform these tasks and are often based on or related to the Kalman filter. Typically Kalman filters assumes that the locations associated with observations are known with certainty. This is reasonable for typical oceanographic measurement methods. Recently, however an alternative and abundant source of data comes from the deployment of ocean sensors on marine animals. This source of data has some attractive properties: unlike traditional oceanographic collection platforms, it is relatively cheap to collect, plentiful, has multiple scientific uses and users, and samples areas of the ocean that are often difficult of costly to sample. However, inherent uncertainty in the location of the observations is a barrier to full utilisation of animal-borne sensor data in data-assimilation schemes. In this article we examine this issue and suggest a simple approximation to explicitly incorporate the location uncertainty, while staying in the scope of Kalman-filter-like methods. The approximation stems from a Taylor-series approximation to elements of the updating equation. PMID:22900005

  19. Development of a new PCR-based assay to detect Anaplasmataceae and the first report of Anaplasma phagocytophilum and Anaplasma platys in cattle from Algeria.

    PubMed

    Dahmani, Mustapha; Davoust, Bernard; Benterki, Mohamed Seghir; Fenollar, Florence; Raoult, Didier; Mediannikov, Oleg

    2015-04-01

    Bovine anaplasmosis is a hemoparasitic disease considered as a major constraint to cattle production in many countries. This pathology is at least partially caused by Anaplasma phagocytophilum, Anaplasma marginale, Anaplasma centrale, and Anaplasma bovis. The global threat and emergence of these species in animals require the reliable identification of these bacteria in animal samples. In this study, we developed a new qPCR tool targeting the 23S rRNA gene for the detection of Anaplasmataceae bacteria. The primers and probe for the qPCR reaction had 100% specificity and could identify at least A. phagocytophilum, A. marginale, A. centrale, Anaplasma ovis, Anaplasma platys, Ehrlichia canis, Ehrlichia ruminantium, Neorickettisa sennetsu, and Neorickettsia risticii. We used this tool to test samples of bovines from Batna (Algeria), an area from which bovine anaplasmosis have never been reported. We identified three genetic variants of A. phagocytophilum, A. platys and Anaplasma sp. "variant 4". This finding should attract the attention of public authorities to assess the involvement of these pathogens in human and animal health. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Detection of border disease virus (BDV) genotype 3 in Italian goat herds.

    PubMed

    Rosamilia, A; Grattarola, C; Caruso, C; Peletto, S; Gobbi, E; Tarello, V; Caroggio, P; Dondo, A; Masoero, L; Acutis, P L

    2014-03-01

    In January 2011, cases of abortion, stillbirth and weak live kids were reported in two goat herds in northern Italy. Samples from 18 kids found dead, 12 fetuses, and two stillborn kids were analyzed for pestivirus antigen using an ELISA kit and a border disease virus (BDV)-specific RT-PCR. Positive results were obtained in six kids and one fetus. Phylogenetic analysis based on 225 bp of the 5'UTR fragment of the BDV genome from positive samples showed that the goats were infected with BDV genotype 3. Serum and blood samples collected from all animals in both herds were analyzed using competitive ELISA to detect p80 antibodies and RT-PCR to detect viraemia. Pestivirus antibodies were detected in 61/67 goats in herd A and in 38/169 in herd B. A persistently infected (PI) goat was found in herd A. The PI animal was submitted to the laboratory for BDV diagnosis with Ag-ELISA, viral isolation, and nested RT-PCR on tissue samples from the spleen, kidney, brain, liver, lung, ileocaecal valve, mesenteric lymph nodes, and skin. All of the tests were positive for BDV in each of the tissues analyzed. The BDV sequence of the PI was identical to BDV sequences found in other positive animals. This is the first description of a BDV PI goat and the first evidence of BDV genotype 3 circulation in Italy. The study raises questions about the real impact this virus has on breeding goats. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. Pesticide exposure on sloths (Bradypus variegatus and Choloepus hoffmanni) in an agricultural landscape of Northeastern Costa Rica.

    PubMed

    Pinnock Branford, Margaret Verónica; de la Cruz, Elba; Solano, Karla; Ramírez, Oscar

    2014-01-01

    Between 2005 and 2008, wild Bradypus variegatus and Choloepus hoffmanni inhabiting an agricultural landscape and captive animals from a rescue center in Northeastern Costa Rica were studied to assess exposure to pesticides. A total of 54 animals were sampled: 42 wild sloths captured at an agricultural landscape and 12 captive animals from a rescue center. Pesticides' active ingredients were determined in three sample matrices: hair, aqueous mixture (paws' wash) and cotton gauze (mouth clean) based on multi-residue gas chromatography methods. Recoveries tests ranged from 73 to 146% and relative standard deviations were less than 20% throughout all the recovery tests. Active ingredients detected in sloths samples were ametryn, chlorothalonil, chlorpyrifos, diazinon, difenoconazole, ethoprophos and thiabendazole. These active ingredients were used in intensive agricultural production for bananas, pineapples and other crops. Blood plasma cholinesterase activity (PChE) was determined by the Ellman method modified for micro plates. Enzyme activity determination was normalized to protein content in the samples according to Bradford method. Wild sloth PChE activity was similar for both species while sloths in captivity showed differences between species. Enzyme activity was significantly lower for two-toed sloths. This study showed that sloths were exposed to pesticides that caused acute and chronic effect in mammals and can also be a threat to other wildlife species. There is a need to better understand the potential effects of exposure to pesticides in sloths and other wild mammal populations, especially those threatened or endangered. More studies in this field must be carried out on the wildlife fauna inhabiting the agricultural landscape and its surroundings.

  2. Detection of Mycoplasma hyopneumoniae by polymerase chain reaction in swine presenting respiratory problems

    PubMed Central

    Yamaguti, M.; Muller, E.E.; Piffer, A.I.; Kich, J.D.; Klein, C.S.; Kuchiishi, S.S.

    2008-01-01

    Since Mycoplasma hyopneumoniae isolation in appropriate media is a difficult task and impractical for daily routine diagnostics, Nested-PCR (N-PCR) techniques are currently used to improve the direct diagnostic sensitivity of Swine Enzootic Pneumonia. In a first experiment, this paper describes a N-PCR technique optimization based on three variables: different sampling sites, sample transport media, and DNA extraction methods, using eight pigs. Based on the optimization results, a second experiment was conducted for testing validity using 40 animals. In conclusion, the obtained results of the N-PCR optimization and validation allow us to recommend this test as a routine monitoring diagnostic method for Mycoplasma hyopneumoniae infection in swine herds. PMID:24031248

  3. Peptidomic Approach to Developing ELISAs for the Determination of Bovine and Porcine Processed Animal Proteins in Feed for Farmed Animals.

    PubMed

    Huet, Anne-Catherine; Charlier, Caroline; Deckers, Elise; Marbaix, Hélène; Raes, Martine; Mauro, Sergio; Delahaut, Philippe; Gillard, Nathalie

    2016-11-30

    The European Commission (EC) wants to reintroduce nonruminant processed animal proteins (PAPs) safely into the feed chain. This would involve replacing the current ban in feed with a species-to-species ban which, in the case of nonruminants, would only prohibit feeding them with proteins from the same species. To enforce such a provision, there is an urgent need for species-specific methods for detecting PAPs from several species in animal feed and in PAPs from other species. Currently, optical microscopy and the polymerase chain reaction are the officially accepted methods, but they have limitations, and alternative methods are needed. We have developed immunoassays using antibodies raised against targets which are not influenced by high temperature and pressure. These targets were identified in a previous study based on an experimental approach. One optimized competitive ELISA detects bovine PAPs at 2% in plant-derived feed. The detection capability demonstrated on blind samples shows a good correlation with mass spectrometry results.

  4. Risk of Escherichia coli O157:H7, Non-O157 Shiga Toxin-Producing Escherichia coli, and Campylobacter spp. in Food Animals and Their Products in Qatar.

    PubMed

    Mohammed, Hussni O; Stipetic, Korana; Salem, Ahmed; McDonough, Patrick; Chang, Yung Fu; Sultan, Ali

    2015-10-01

    Escherichia coli O157:H7, non-O157 E. coli, and Campylobacter spp. are among the top-ranked pathogens that threaten the safety of food supply systems around the world. The associated risks and predisposing factors were investigated in a dynamic animal population using a repeat-cross-sectional study design. Animal and environmental samples were collected from dairy and camel farms, chicken processing plants, and abattoirs and analyzed for the presence of these pathogens using a combination of bacterial enrichment and real-time PCR tests without culture confirmation. Data on putative risk factors were also collected and analyzed. E. coli O157:H7 was detected by PCR at higher levels in sheep and camel feces than in cattle feces (odds ratios [OR], 6.8 and 21.1, respectively). Although the genes indicating E. coli O157:H7 were detected at a relatively higher rate (4.3%) in fecal samples from dairy cattle, they were less common in milk and udder swabs from the same animals (1 and 2%, respectively). Among the food adulterants, E. coli O103 was more common in cattle fecal samples, whereas O26 was more common in sheep feces and O45 in camel feces compared with cattle (OR, 2.6 and 3.1, respectively). The occurrence of E. coli in the targeted populations differed by the type of sample and season of the year. Campylobacter jejuni and Campylobacter coli were more common in sheep and camel feces than in cattle feces. Most of the survey and surveillance of E. coli focused on serogroup O157 as a potential foodborne hazard; however, based on the PCR results, non-O157 Shiga toxin-producing E. coli serotypes appeared to be more common, and efforts should be made to include them in food safety programs.

  5. Molecular identification of trypanosomatids in wild animals.

    PubMed

    Tenório, M S; Oliveira e Sousa, L; Alves-Martin, M F; Paixão, M S; Rodrigues, M V; Starke-Buzetti, W A; Araújo Junior, J P; Lucheis, S B

    2014-06-16

    Diverse wild animal species can be reservoirs of zoonotic flagellate parasites, which can cause pathologic Chagas disease. The present study aimed to detect the natural occurrence of flagellate parasites through direct microscopic examination of the parasites in blood samples and through PCR of whole blood and blood culture (haemoculture) samples from 38 captive and 65 free-living wild animals in the Centre for Conservation of Wild Fauna (CCWF), an area endemic for leishmaniasis. For this study, PCR was accomplished using primers for the ribosomal region (ITS-1) of the flagellate parasites. The amplified fragments were cloned and sequenced to identify DNA of the Trypanosomatid parasite species, observed in blood cultures from 3.9% (04/103) of the animals. Through these techniques, Trypanosoma cruzi was identified in haemoculture samples of the following three free-living species: common agouti (Dasyprocta aguti), white-eared opossum (Didelphis albiventris), and nine-banded armadillo (Dasypus novemcinctus). Furthermore, Trypanosoma minasense was identified in whole blood samples from 01 (0.9%) captive animal (black howler monkey-Alouatta caraya). These results demonstrated the first report of T. cruzi isolation in wild species from the CCWF using blood culture, which can be applied in addition to molecular tools for epidemiological studies and to identify trypanosomatids in wild animals. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Detection of Different DNA Animal Species in Commercial Candy Products.

    PubMed

    Muñoz-Colmenero, Marta; Martínez, Jose Luis; Roca, Agustín; Garcia-Vazquez, Eva

    2016-03-01

    Candy products are consumed all across the world, but there is not much information about their composition. In this study we have used a DNA-based approach for determining the animal species occurring in 40 commercial candies of different types. We extracted DNA and performed PCR amplification, cloning and sequencing for obtaining species-informative DNA sequences. Eight species were identified including fish (hake and anchovy) in 22% of the products analyzed. Bovine and porcine were the most abundant appearing in 27 samples each one. Most products contained a mixture of species. Marshmallows (7), jelly-types, and gummies (20) contained a significantly higher number of species than hard candies (9). We demonstrated the presence of DNA animal species in candy product which allow consumers to make choices and prevent allergic reaction. © 2016 Institute of Food Technologists®

  7. Anthropogenic impact on the presence of L. monocytogenes in soil, fruits, and vegetables.

    PubMed

    Szymczak, Barbara; Szymczak, Mariusz; Sawicki, Wojciech; Dąbrowski, Waldemar

    2014-01-01

    The aim of this study was to determine the prevalence of Listeria sp. and Listeria monocytogenes in soil samples with reference to type of fertilizers (natural and artificial) and distance from places intensively exploited by men, as well as to determine the relationship between the presence of L. monocytogenes in the soil and in fruits and vegetables. The examined 1,000 soil samples originated from 15 different areas, whilst 140 samples of fruits and 210 samples of vegetables were collected from those areas. L. monocytogenes was isolated only from 5.5 % of all soil samples coming exclusively from meadows intensively grazed by cattle (27.8 %) and areas near food processing plants (25 %) and wild animal forests (24 %). Listeria sp. and L. monocytogenes were not present on artificially fertilized areas and wastelands. L. monocytogenes was detected in 10 % of samples of strawberry, 15 % of potato samples, and 5 % of parsley samples. Our data indicate that Listeria spp. and particularly L. monocytogenes were found in the soil from (1) arable lands fertilized with manure, (2) pasture (the land fertilized with feces of domestic animals), and (3) forests (again, the land fertilized with feces of animals, not domestic but wild). The bacteria were not detected in the soil samples collected at (1) artificially fertilized arable lands and (2) wastelands (the lands that were not fertilized with manure or animal feces). Moreover, a correlation was determined in the presence of L. monocytogenes between soil samples and samples of the examined fruits and vegetables.

  8. Incidence of Nontyphoidal Salmonella in Food-Producing Animals, Animal Feed, and the Associated Environment in South Africa, 2012-2014.

    PubMed

    Magwedere, Kudakwashe; Rauff, Dionne; De Klerk, Grietjie; Keddy, Karen H; Dziva, Francis

    2015-11-01

    Nontyphoidal salmonellosis continues to pose a global threat to human health, primarily by causing food-borne illnesses, and food-producing animals are the principal reservoirs of many pathogenic serovars. To identify key control points and generate information that may enable future estimation of the transmission routes between the environment, animals, and humans, we examined data on Salmonella isolates in South Africa. Samples were obtained from livestock and poultry on farms, meat at abattoirs, raw materials at feed mills, animal feed, and environmental sources (eg, poultry houses, abattoirs, feed mills, water) from 2012 to 2014 in compliance with each establishment's protocols conforming to International Organization for Standardization (ISO) (ISO/TS 17728, ISO 18593:2004 and ISO 17604:2003) standards. Isolation and serotyping of Salmonella were performed according to the scope of accreditation of the respective laboratories conforming to ISO/IEC 17025:2005 standard techniques. Salmonella was isolated from 9031 of 180 298 (5.0%) samples, and these isolates were distributed among 188 different serovars. Salmonella Enteritidis was the most frequent isolate, with 1944 of 180 298 (21.5%) originating from poultry on farms, poultry meat, and poultry houses, followed by Salmonella Havana, with 677 of 180 298 (7.5%), mostly from environmental samples. Serovars that are uncommonly associated with human disease (Salmonella Idikan, Salmonella Salford, and Salmonella Brancaster) were isolated at higher frequencies than Salmonella Typhimurium, a common cause of human illness. Environmental samples accounted for 3869 of 9031 (42.8%) samples positive for Salmonella. We describe the frequent isolation of Salmonella of a wide variety of serovars, from an array of animal feeds, food animals, and food animal environment. As prevention of human salmonellosis requires the effective control of Salmonella in food animals, these data can be used to facilitate Salmonella control in food animals and thereby prevent human infections. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  9. Molecular investigation and phylogeny of Anaplasmataceae species infecting domestic animals and ticks in Corsica, France.

    PubMed

    Dahmani, Mustapha; Davoust, Bernard; Tahir, Djamel; Raoult, Didier; Fenollar, Florence; Mediannikov, Oleg

    2017-06-23

    Corsica is a French island situated in the Mediterranean Sea. The island provides suitable natural conditions to study disease ecology, especially tick-borne diseases and emerging diseases in animals and ticks. The family Anaplasmataceae is a member of the order Rickettsiales; it includes the genera Anaplasma, Ehrlichia, Neorickettsia and Wolbachia. Anaplasmosis and ehrlichiosis traditionally refer to diseases caused by obligate intracellular bacteria of the genera Anaplasma and Ehrlichia. The aim of this study was to identify and estimate the prevalence of Anaplasmataceae species infecting domestic animals and ticks in Corsica. In this study, 458 blood samples from sheep, cattle, horses, goats, dogs, and 123 ticks removed from cattle, were collected in Corsica. Quantitative real-time PCR screening and genetic characterisation of Anaplasmataceae bacteria were based on the 23S rRNA, rpoB and groEl genes. Two tick species were collected in the present study: Rhipicephalus bursa (118) and Hyalomma marginatum marginatum (5). Molecular investigation showed that 32.1% (147/458) of blood samples were positive for Anaplasmataceae infection. Anaplasma ovis was identified in 42.3% (93/220) of sheep. Anaplasma marginale was amplified from 100% (12/12) of cattle and two R. bursa (2/123). Several potentially new species were also identified: Anaplasma cf. ovis, "Candidatus Anaplasma corsicanum", "Candidatus Anaplasma mediterraneum" were amplified from 17.3% (38/220) of sheep, and Anaplasma sp. marginale-like was amplified from 80% (4/5) of goats. Finally, one R. bursa tick was found to harbour the DNA of E. canis. All samples from horses and dogs were negative for Anaplasmataceae infection. To our knowledge, this study is the first epidemiological survey on Anaplasmataceae species infecting animals and ticks in Corsica and contributes toward the identification of current Anaplasmataceae species circulating in Corsica.

  10. Near-infrared microscopic methods for the detection and quantification of processed by-products of animal origin

    NASA Astrophysics Data System (ADS)

    Abbas, O.; Fernández Pierna, J. A.; Dardenne, P.; Baeten, V.

    2010-04-01

    Since the BSE crisis, researches concern mainly the detection, identification, and quantification of meat and bone meal with an important focus on the development of new analytical methods. Microscopic based spectroscopy methods (NIR microscopy - NIRM or/and NIR hyperspectral imaging) have been proposed as complementary methods to the official method; the optical microscopy. NIR spectroscopy offers the advantage of being rapid, accurate and independent of human analyst skills. The combination of an NIR detector and a microscope or a camera allows the collection of high quality spectra for small feed particles having a size larger than 50 μm. Several studies undertaken have demonstrated the clear potential of NIR microscopic methods for the detection of animal particles in both raw and sediment fractions. Samples are sieved and only the gross fraction (superior than 250 μm) is investigated. Proposed methodologies have been developed to assure, with an acceptable level of confidence (95%), the detection of at least one animal particle when a feed sample is adulterated at a level of 0.1%. NIRM and NIR hyperspectral imaging are running under accreditation ISO 17025 since 2005 at CRA-W. A quantitative NIRM approach has been developed in order to fulfill the new requirements of the European commission policies. The capacities of NIRM method have been improved; only the raw fraction is analyzed, both the gross and the fine fractions of the samples are considered, and the acquisition parameters are optimized (the aperture, the gap, and the composition of the animal feed). A mapping method for a faster collection of spectra is also developed. The aim of this work is to show the new advances in the analytical methods developed in the frame of the feed ban applied in Europe.

  11. Experiment K-7-29: Connective Tissue Studies. Part 3; Rodent Tissue Repair: Skeletal Muscle

    NASA Technical Reports Server (NTRS)

    Stauber, W.; Fritz, V. K.; Burkovskaya, T. E.; Ilyina-Kakueva, E. I.

    1994-01-01

    Myofiber injury-repair was studied in the rat gastrocnemius following a crush injury to the lower leg prior to flight in order to understand if the regenerative responses of muscles are altered by the lack of gravitational forces during Cosmos 2044 flight. After 14 days of flight, the gastrocnemius muscle was removed from the 5 injured flight rodents and various Earth-based treatment groups for comparison. The Earth-based animals consisted of three groups of five rats with injured muscles from a simulated, tail-suspended, and vivarium as well as an uninjured basal group. The gastrocnemius muscle from each was evaluated by histochemical and immunohistochemical techniques to document myofiber, vascular, and connective tissue alterations following injury. In general the repair process was somewhat similar in all injured muscle samples with regard to extracellular matrix organization and myofiber regeneration. Small and large myofibers were present with a newly organized extracellular matrix indicative of myogenesis and muscle regeneration. In the tail-suspended animals, a more complete repair was observed with no enlarged area of non-muscle cells or matrix material visible. In contrast, the muscle samples from the flight animals were less well differentiated with more macrophages and blood vessels in the repair region but small myofibers and proteoglycans, nevertheless, were in their usual configuration. Thus, myofiber repair did vary in muscles from the different groups, but for the most part, resulted in functional muscle tissue.

  12. Growth of bacterial phytopathogens in animal manures.

    PubMed

    Sledz, Wojciech; Zoledowska, Sabina; Motyka, Agata; Kadziński, Leszek; Banecki, Bogdan

    2017-01-01

    Animal manures are routinely applied to agricultural lands to improve crop yield, but the possibility to spread bacterial phytopathogens through field fertilization has not been considered yet. We monitored 49 cattle, horse, swine, sheep or chicken manure samples collected in 14 Polish voivodeships for the most important plant pathogenic bacteria - Ralstonia solanacearum (Rsol), Xanthomonas campestris pv. campestris (Xcc), Pectobacterium carotovorum subsp. carotovorum (Pcc), Pectobacterium atrosepticum (Pba), Erwinia amylovora (Eam), Clavibacter michiganensis subsp. sepedonicus (Cms) and Dickeya sp. (Dsp). All of the tested animal fertilizers were free of these pathogens. Subsequently, the growth dynamics of Pba, Pcc, Rsol, and Xcc in cattle, horse, swine, sheep and chicken manures sterilized either by autoclaving or filtration was evaluated. The investigated phytopathogens did not exhibit any growth in the poultry manure. However, the manure filtrates originating from other animals were suitable for microbial growth, which resulted in the optical density change of 0.03-0.22 reached within 26 h (48 h Rsol, 120 h Xcc), depending on bacterial species and the manure source. Pcc and Pba multiplied most efficiently in the cattle manure filtrate. These bacteria grew faster than Rsol and Xcc in all the tested manure samples, both the filtrates and the autoclaved semi-solid ones. Though the growth dynamics of investigated strains in different animal fertilizers was unequal, all of the tested bacterial plant pathogens were proven to use cattle, horse, swine and sheep manures as the sources of nutrients. These findings may contribute to further research on the alternative routes of spread of bacterial phytopathogens, especially because of the fact that the control of pectionolytic bacteria is only based on preventive methods.

  13. Human paleodiet and animal utilization strategies during the Bronze Age in northwest Yunnan Province, southwest China.

    PubMed

    Ren, Lele; Li, Xin; Kang, Lihong; Brunson, Katherine; Liu, Honggao; Dong, Weimiao; Li, Haiming; Min, Rui; Liu, Xu; Dong, Guanghui

    2017-01-01

    Reconstructing ancient diets and the use of animals and plants augment our understanding of how humans adapted to different environments. Yunnan Province in southwest China is ecologically and environmentally diverse. During the Neolithic and Bronze Age periods, this region was occupied by a variety of local culture groups with diverse subsistence systems and material culture. In this paper, we obtained carbon (δ13C) and nitrogen (δ15N) isotopic ratios from human and faunal remains in order to reconstruct human paleodiets and strategies for animal exploitation at the Bronze Age site of Shilinggang (ca. 2500 Cal BP) in northwest Yunnan Province. The δ13C results for human samples from Shilinggang demonstrate that people's diets were mainly dominated by C3-based foodstuffs, probably due to both direct consumption of C3 food and as a result of C3 foddering of consumed animals. Auxiliary C4 food signals can also be detected. High δ15N values indicate that meat was an important component of the diet. Analysis of faunal samples indicates that people primarily fed pigs and dogs with human food waste, while sheep/goats and cattle were foddered with other food sources. We compare stable isotope and archaeobotanical data from Shilinggang with data from other Bronze Age sites in Yunnan to explore potential regional variation in subsistence strategies. Our work suggests that people adopted different animal utilization and subsistence strategies in different parts of Yunnan during the Bronze Age period, probably as local adaptations to the highly diversified and isolated environments in the region.

  14. THAILAND AND LAO PDR: FOCUS ON HOOKWORM AND THREADWORM.

    PubMed

    Niamnuy, Nunthawadee; Kaewthamasorn, Morakot; Congpuong, Kanungnit; Phaytanavanh, Bounthanh; Lohsoonthorn, Vitool

    2016-09-01

    Hookworm and threadworm infections are major public health problems in developing countries. A cross sectional study comprising 843 participants (346 males and 497 females) was conducted in three populations: i) Thai residents (TR) of Ubon Ratchathani Province, Thailand; ii) Laotian immigrant workers (LI) in the same province; and iii) Laotian residents (LR) in Champasak Province, Lao PDR. Participants were interviewed based on a structured questionnaire regarding their health status. Stool samples from participants and 300 samples from domestic animals (277 dogs and 23 cats) living in the participants households were collected and examined for parasitic infection using a formalin-ether concentration and a Harada-Mori filter paper culture techniques. Approximately one-third of TR and LI populations and domestic animals in Thailand were positive for parasitic infections, while almost half of LR population and domestic animals were positive. We confirmed by PCR and DNA sequencing a case of Ancylostoma ceylanicum infection in a Thai man. We also observed infections of other parasites, such as Taenia spp and Opisthorchis viverrini. Multivariate analysis indicated that risk factors for hookworm infection were population group and walking barefoot. Factors associated with threadworm infection were population group, adult male, lack of previous antiparasitic treatment and of knowledge of parasitic infection, and failure to wash hands after contact with domestic animals. Our results highlight the high prevalence of both hookworm and threadworm infections especially among LI population and domestic animals in both countries. Our findings emphasize the need for public health intervention to control the spread of parasitic infections in Thailand and Lao PDR.

  15. Rapid shape detection signals in area V4

    PubMed Central

    Weiner, Katherine F.; Ghose, Geoffrey M.

    2014-01-01

    Vision in foveate animals is an active process that requires rapid and constant decision-making. For example, when a new object appears in the visual field, we can quickly decide to inspect it by directing our eyes to the object's location. We studied the contribution of primate area V4 to these types of rapid foveation decisions. Animals performed a reaction time task that required them to report when any shape appeared within a peripherally-located noisy stimulus by making a saccade to the stimulus location. We found that about half of the randomly sampled V4 neurons not only rapidly and precisely represented the appearance of this shape, but they were also predictive of the animal's saccades. A neuron's ability to predict the animal's saccades was not related to the specificity with which the cell represented a single type of shape but rather to its ability to signal whether any shape was present. This relationship between sensory sensitivity and behavioral predictiveness was not due to global effects such as alertness, as it was equally likely to be observed for cells with increases and decreases in firing rate. Careful analysis of the timescales of reliability in these neurons implies that they reflect both feedforward and feedback shape detecting processes. In approximately 7% of our recorded sample, individual neurons were able to predict both the delay and precision of the animal's shape detection performance. This suggests that a subset of V4 neurons may have been directly and causally contributing to task performance and that area V4 likely plays a critical role in guiding rapid, form-based foveation decisions. PMID:25278828

  16. Human paleodiet and animal utilization strategies during the Bronze Age in northwest Yunnan Province, southwest China

    PubMed Central

    Ren, Lele; Li, Xin; Kang, Lihong; Brunson, Katherine; Liu, Honggao; Dong, Weimiao; Li, Haiming; Min, Rui; Liu, Xu

    2017-01-01

    Reconstructing ancient diets and the use of animals and plants augment our understanding of how humans adapted to different environments. Yunnan Province in southwest China is ecologically and environmentally diverse. During the Neolithic and Bronze Age periods, this region was occupied by a variety of local culture groups with diverse subsistence systems and material culture. In this paper, we obtained carbon (δ13C) and nitrogen (δ15N) isotopic ratios from human and faunal remains in order to reconstruct human paleodiets and strategies for animal exploitation at the Bronze Age site of Shilinggang (ca. 2500 Cal BP) in northwest Yunnan Province. The δ13C results for human samples from Shilinggang demonstrate that people’s diets were mainly dominated by C3-based foodstuffs, probably due to both direct consumption of C3 food and as a result of C3 foddering of consumed animals. Auxiliary C4 food signals can also be detected. High δ15N values indicate that meat was an important component of the diet. Analysis of faunal samples indicates that people primarily fed pigs and dogs with human food waste, while sheep/goats and cattle were foddered with other food sources. We compare stable isotope and archaeobotanical data from Shilinggang with data from other Bronze Age sites in Yunnan to explore potential regional variation in subsistence strategies. Our work suggests that people adopted different animal utilization and subsistence strategies in different parts of Yunnan during the Bronze Age period, probably as local adaptations to the highly diversified and isolated environments in the region. PMID:28531221

  17. Differences in Preschool Children's Conceptual Strategies When Thinking about Animate Entities and Artifacts.

    ERIC Educational Resources Information Center

    Blanchet, Nicole; Dunham, Philip J.; Dunham, Frances

    2001-01-01

    Preschoolers viewed stimulus sets comprised of a sample picture and three types of matches and were asked to choose a match that "went with" each sample. Children's choices indicated that a shift occurs between 3 and 4 years of age from a taxonomic bias to a thematic bias. Animate sample stimuli enhanced children's tendency to adopt…

  18. Mycotoxins and Mycotoxigenic Fungi in Poultry Feed for Food-Producing Animals

    PubMed Central

    Greco, Mariana Vanesa; Rico Golba, Silvia Laura; Pardo, Alejandro Guillermo; Pose, Graciela Noemí

    2014-01-01

    Moulds are capable of reducing the nutritional value of feedstuff as well as elaborating several mycotoxins. Mycotoxin-contaminated feed has adverse effects on animal health and productivity. Also, mycotoxins may be carried over into meat and eggs when poultry are fed with contaminated feed. In a point prevalence study feedstuff used for poultry nutrition in Argentina was analyzed for fungal flora, natural incidence of selected mycotoxins, and nutritional quality. Ten mould genera were recovered, six of them known to be mycotoxigenic. More than 28 species were determined. Fumonisins were detected in all the samples (median 1,750 ppb). Forty-four out of 49 samples (90%) were contaminated with DON (median 222 ppb) and OTA (median 5 ppb). Also, 44 out of 49 samples were contaminated with aflatoxins (median 2.685 ppb), 42 samples (86%) with ZEA (median 50 ppb), and 38 samples (78%) with T2-toxin (median 50 ppb). Ninety percent of the samples had at least one type of nutritional deficiency. This study indicates the need for continuous assessment of the mycological status of animal feed production, in order to feed animals for optimal performance ensuring food safety. PMID:25126610

  19. Occurrence and molecular characterization of Cryptosporidium spp. isolated from domestic animals in a rural area surrounding Atlantic dry forest fragments in Teodoro Sampaio municipality, State of São Paulo, Brazil.

    PubMed

    Sevá, Anaiá da Paixão; Funada, Mikaela Renata; Souza, Sheila de Oliveira; Nava, Alessandra; Richtzenhain, Leonardo José; Soares, Rodrigo Martins

    2010-01-01

    The aim of this study was to assess the occurrence of Cryptosporidium in domestic animals in rural properties surrounding rain forest fragments within the municipality of Teodoro Sampaio, southeastern Brazil. Conventional sucrose flotation method followed by molecular characterization of the parasites by sequencing PCR products amplified from SSU rRNA gene were used. Stool samples were collected from domestic animals raised as pets and livestock in all rural properties surrounding three forest fragments. Samples from cattle (197), equine (63), pigs (25), sheep (11), and dogs (28) were collected from 98 rural properties. The frequency of occurrence of Cryptosporidium within each animal species was 3.0% (6/197) among cattle and 10.7% (3/28) among dogs. Cryptosporidium was not detected in stool samples from equine, sheep, and pigs. All sequences obtained from the six samples of calves showed molecular identity with Cryptosporidium andersoni while all sequences from dog samples were similar to C. canis. The frequency of occurrence of Cryptosporidium in these domestic animal species was low. The absence of C. parvum in the present study suggests that the zoonotic cycle of cryptosporidiosis may not be relevant in the region studied. The presence of Cryptosporidium species seldom described in humans may be, otherwise, important for the wild fauna as these animals are a source of infection and dissemination of this protozoan to other animal species. The impact and magnitude of infection by C. andersoni in wild ruminants and C. canis in wild canids have to be assessed in future studies to better understand the actual importance of these species in this region.

  20. Point of Care Tuberculosis Sero-Diagnosis Kit for Wild Animals: Combination of Proteins for Improving the Diagnostic Sensitivity and Specificity.

    PubMed

    Veerasami, Maroudam; Venkataraman, K; Karuppannan, Chitra; Shanmugam, Arun Attur; Prudhvi, Mallepaddi Chand; Holder, Thomas; Rathnagiri, Polavarapu; Arunmozhivarman, K; Raj, Gopal Dhinakar; Vordermeier, Martin; Mohana Subramanian, B

    2018-03-01

    Tuberculosis is a significant problem globally for domestic animals as well as captive and free ranging wild life. Rapid point of care (POC) serology kits are well suited for the diagnosis of TB in wild animals. However, wild animals are invariably exposed to environmental non-pathogenic mycobacterium species with the development of cross reacting antibodies. In the present study, POC TB diagnosis kit was developed using a combination of pathogenic Mycobacteria specific recombinant antigens and purified protein derivatives of pathogenic and non-pathogenic Mycobacteria . To benchmark the TB antibody detection kit, particularly in respect to specificity which could not be determined in wildlife due to the lack of samples from confirmed uninfected animals, we first tested well-characterized sera from 100 M. bovis infected and 100 uninfected cattle. Then we investigated the kit's performance using sera samples from wildlife, namely Sloth Bears (n = 74), Elephants (n = 9), Cervidae (n = 14), Felidae (n = 21), Cape buffalo (n = 2), Wild bear (n = 1) and Wild dog (n = 1).In cattle, a sensitivity of 81% and a specificity of 90% were obtained. The diagnostic sensitivity of the kit was 94% when the kit was tested using known TB positive sloth bear sera samples. 47.4% of the in-contact sloth bears turned seropositive using the rapid POC TB diagnostic kit. Seropositivity in other wild animals was 25% when the sera samples were tested using the kit. A point of care TB sero-diagnostic kit with the combination of proteins was developed and the kit was validated using the sera samples of wild animals.

  1. Development of Cross-Assembly Phage PCR-Based Methods ...

    EPA Pesticide Factsheets

    Technologies that can characterize human fecal pollution in environmental waters offer many advantages over traditional general indicator approaches. However, many human-associated methods cross-react with non-human animal sources and lack suitable sensitivity for fecal source identification applications. The genome of a newly discovered bacteriophage (~97 kbp), the Cross-Assembly phage or “crAssphage”, assembled from a human gut metagenome DNA sequence library is predicted to be both highly abundant and predominately occur in human feces suggesting that this double stranded DNA virus may be an ideal human fecal pollution indicator. We report the development of two human-associated crAssphage endpoint PCR methods (crAss056 and crAss064). A shotgun strategy was employed where 384 candidate primers were designed to cover ~41 kbp of the crAssphage genome deemed favorable for method development based on a series of bioinformatics analyses. Candidate primers were subjected to three rounds of testing to evaluate assay optimization, specificity, limit of detection (LOD95), geographic variability, and performance in environmental water samples. The top two performing candidate primer sets exhibited 100% specificity (n = 70 individual samples from 8 different animal species), >90% sensitivity (n = 10 raw sewage samples from different geographic locations), LOD95 of 0.01 ng/µL of total DNA per reaction, and successfully detected human fecal pollution in impaired envi

  2. Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains.

    PubMed

    Wilkes, Rebecca P; Sanchez, Elena; Riley, Matthew C; Kennedy, Melissa A

    2014-01-01

    Canine distemper virus (CDV) remains a common cause of infectious disease in dogs, particularly in high-density housing situations such as shelters. Vaccination of all dogs against CDV is recommended at the time of admission to animal shelters and many use a modified live virus (MLV) vaccine. From a diagnostic standpoint for dogs with suspected CDV infection, this is problematic because highly sensitive diagnostic real-time reverse transcription polymerase chain reaction (RT-PCR) tests are able to detect MLV virus in clinical samples. Real-time PCR can be used to quantitate amount of virus shedding and can differentiate vaccine strains from wild-type strains when shedding is high. However, differentiation by quantitation is not possible in vaccinated animals during acute infection, when shedding is low and could be mistaken for low level vaccine virus shedding. While there are gel-based RT-PCR assays for differentiation of vaccine strains from field strains based on sequence differences, the sensitivity of these assays is unable to match that of the real-time RT-PCR assay currently used in the authors' laboratory. Therefore, a real-time RT-PCR assay was developed that detects CDV MLV vaccine strains and distinguishes them from wild-type strains based on nucleotide sequence differences, rather than the amount of viral RNA in the sample. The test is highly sensitive, with detection of as few as 5 virus genomic copies (corresponding to 10(-1) TCID(50)). Sequencing of the DNA real-time products also allows phylogenetic differentiation of the wild-type strains. This test will aid diagnosis during outbreaks of CDV in recently vaccinated animals.

  3. Robust Abundance Estimation in Animal Abundance Surveys with Imperfect Detection

    EPA Science Inventory

    Surveys of animal abundance are central to the conservation and management of living natural resources. However, detection uncertainty complicates the sampling process of many species. One sampling method employed to deal with this problem is depletion (or removal) surveys in whi...

  4. Robust Abundance Estimation in Animal Surveys with Imperfect Detection

    EPA Science Inventory

    Surveys of animal abundance are central to the conservation and management of living natural resources. However, detection uncertainty complicates the sampling process of many species. One sampling method employed to deal with this problem is depletion (or removal) surveys in whi...

  5. [Leptospirosis in animal reproduction: III. Role of the hardjo serovar in bovine leptospirosis in Rio de Janeiro, Brazil].

    PubMed

    Lilenbaum, W; Dos Santos, M R

    1995-01-01

    Four hundred and five serum samples were drawn from cows with reproductive problems which were not vaccinated against leptospirosis from 21 dairy farms. Three distinct geographic regions were determined and the farms were also classified considering the production system, based on technological, zootechnical and sanitary resources. A total of 277 positive reactions were observed, corresponding to 68.39% of the samples. The predominant serovar was hardjo, reactive on 85 samples (20.98%), predominant on nine farms and observed on 17 farms (80.95%). It was observed the predominance of hardjo in all studied regions and on properties classified as type "A" (22 samples) and type "B" (49 samples). The role of this serovar on bovine leptospirosis in Brazil compared with other countries is discussed.

  6. Project BETA: Biological Education Through Animals.

    ERIC Educational Resources Information Center

    Abramson, Charles I.; Wallisch, Kristy; Huss, Jeanine M.; Payne, Delissa

    1999-01-01

    Describes a program in which biology students study animals in pet shops to learn about animal behavior. Lists general guidelines for starting a partnership and presents two sample student activities. (WRM)

  7. Zoonotic pathogens isolated from wild animals and environmental samples at two California wildlife hospitals.

    PubMed

    Siembieda, Jennifer L; Miller, Woutrina A; Byrne, Barbara A; Ziccardi, Michael H; Anderson, Nancy; Chouicha, Nadira; Sandrock, Christian E; Johnson, Christine K

    2011-03-15

    To determine types and estimate prevalence of potentially zoonotic enteric pathogens shed by wild animals admitted to either of 2 wildlife hospitals and to characterize distribution of these pathogens and of aerobic bacteria in a hospital environment. Cross-sectional study. Fecal samples from 338 animals in 2 wildlife hospitals and environmental samples from 1 wildlife hospital. Fecal samples were collected within 24 hours of hospital admission. Environmental samples were collected from air and surfaces. Samples were tested for zoonotic pathogens via culture techniques and biochemical analyses. Prevalence of pathogen shedding was compared among species groups, ages, sexes, and seasons. Bacterial counts were determined for environmental samples. Campylobacter spp, Vibrio spp, Salmonella spp, Giardia spp, and Cryptosporidium spp (alone or in combination) were detected in 105 of 338 (31%) fecal samples. Campylobacter spp were isolated only from birds. Juvenile passerines were more likely to shed Campylobacter spp than were adults; prevalence increased among juvenile passerines during summer. Non-O1 serotypes of Vibrio cholerae were isolated from birds; during an oil-spill response, 9 of 10 seabirds screened were shedding this pathogen, which was also detected in environmental samples. Salmonella spp and Giardia spp were isolated from birds and mammals; Cryptosporidium spp were isolated from mammals only. Floors of animal rooms had higher bacterial counts than did floors with only human traffic. Potentially zoonotic enteric pathogens were identified in samples from several species admitted to wildlife hospitals, indicating potential for transmission if prevention is not practiced.

  8. Dead or alive: animal sampling during Ebola hemorrhagic fever outbreaks in humans

    PubMed Central

    Olson, Sarah H.; Reed, Patricia; Cameron, Kenneth N.; Ssebide, Benard J.; Johnson, Christine K.; Morse, Stephen S.; Karesh, William B.; Mazet, Jonna A. K.; Joly, Damien O.

    2012-01-01

    There are currently no widely accepted animal surveillance guidelines for human Ebola hemorrhagic fever (EHF) outbreak investigations to identify potential sources of Ebolavirus (EBOV) spillover into humans and other animals. Animal field surveillance during and following an outbreak has several purposes, from helping identify the specific animal source of a human case to guiding control activities by describing the spatial and temporal distribution of wild circulating EBOV, informing public health efforts, and contributing to broader EHF research questions. Since 1976, researchers have sampled over 10,000 individual vertebrates from areas associated with human EHF outbreaks and tested for EBOV or antibodies. Using field surveillance data associated with EHF outbreaks, this review provides guidance on animal sampling for resource-limited outbreak situations, target species, and in some cases which diagnostics should be prioritized to rapidly assess the presence of EBOV in animal reservoirs. In brief, EBOV detection was 32.7% (18/55) for carcasses (animals found dead) and 0.2% (13/5309) for live captured animals. Our review indicates that for the purposes of identifying potential sources of transmission from animals to humans and isolating suspected virus in an animal in outbreak situations, (1) surveillance of free-ranging non-human primate mortality and morbidity should be a priority, (2) any wildlife morbidity or mortality events should be investigated and may hold the most promise for locating virus or viral genome sequences, (3) surveillance of some bat species is worthwhile to isolate and detect evidence of exposure, and (4) morbidity, mortality, and serology studies of domestic animals should prioritize dogs and pigs and include testing for virus and previous exposure. PMID:22558004

  9. Seroprevalence and risk factors associated with bovine brucellosis in the Potohar Plateau, Pakistan.

    PubMed

    Ali, Shahzad; Akhter, Shamim; Neubauer, Heinrich; Melzer, Falk; Khan, Iahtasham; Abatih, Emmanuel Nji; El-Adawy, Hosny; Irfan, Muhammad; Muhammad, Ali; Akbar, Muhammad Waqas; Umar, Sajid; Ali, Qurban; Iqbal, Muhammad Naeem; Mahmood, Abid; Ahmed, Haroon

    2017-01-28

    The seroprevalence and risk factors of bovine brucellosis were studied at animal and herd level using a combination of culture, serological and molecular methods. The study was conducted in 253 randomly selected cattle herds of the Potohar plateau, Pakistan from which a total of 2709 serum (1462 cattle and 1247 buffaloes) and 2330 milk (1168 cattle and 1162 buffaloes) samples were collected. Data on risk factors associated with seroprevalence of brucellosis were collected through interviews using questionnaires. Univariable and multivariable random effects logistic regression models were used for identifying important risk factors at animal and herd levels. One hundred and seventy (6.3%) samples and 47 (18.6%) herds were seropositive for brucellosis by Rose Bengal Plate test. Variations in seroprevalence were observed across the different sampling sites. At animal level, sex, species and stock replacement were found to be potential risk factors for brucellosis. At herd level, herd size (≥9 animals) and insemination method used were important risk factors. The presence of Brucella DNA was confirmed with a real-time polymerase chain reaction assay (qRT-PCR) in 52.4% out of 170 serological positive samples. In total, 156 (6.7%) milk samples were positive by milk ring test. B. abortus biovar 1 was cultured from 5 positive milk samples. This study shows that the seroprevalence of bovine brucellosis is high in some regions in Pakistan. Prevalence was associated with herd size, abortion history, insemination methods used, age, sex and stock replacement methods. The infected animal may act as source of infection for other animals and for humans. The development of control strategies for bovine brucellosis through implementation of continuous surveillance and education programs in Pakistan is warranted.

  10. [Sero-epidemiologic investigation on tick-borne diseases of humans and domestic animals in Zhejiang province].

    PubMed

    Chai, Cheng-liang; Lu, Qun-ying; Sun, Ji-min; Jiang, Li-ping; Ling, Feng; Zhang, Li-juan; Zheng, Shou-gui; Zhang, Hong; Ge, Jun-hua

    2010-10-01

    To investigate the seroprevalence of tick-borne diseases in humans and domestic animals from rural areas of Zhejiang province. Anji county, Jindong district and Tiantai county were selected for samples collection according to their geographic locations and historical prevalence of tick-borne diseases. Blood samples of humans and domestic animals were collected in the three sites. An indirect immuno-fluorescent antibody test was used to determine the presence of IgG antibodies of Rickettsiae heilongjiangii, Orientia tsutsugamushi, R. typhi, Anaplasma phagocytosis, Ehrlichia chaffeensis, Bartonella, R. hainan and Coxiella burnetii in these samples. Six hundred and eighty-three blood samples including 579 from humans and 104 from domestic animals (53 from cattles and 51 from sheep) were collected from the three sites. Antibody positive rates of Orientia tsutsugamushi, R. typhi, Ehrlichia chaffeensis and Coxiella burnetii were significantly different between these sites. IgG from all the 8 pathogens were detected in samples from humans. It was found that the sero-prevalence rates of R. typhi, Bartonella and C. burnetii (20.7%, 10.9%, 5.5%) of adults were higher than those of other Rickettsiae under investigation. The seroprevalence of R. typhi increased along with age. IgG from the 7 pathogens were detected in samples from domestic animals except for Anaplasma phagocytosis. The sero-prevalence rates of R. typhi, Bartonella and R. hainan (69.2%, 51.0%, 22.1%) of adults were higher than those of other Rickettsiae investigated. Tick-borne diseases did spread widely in humans and domestic animals from different rural areas of Zhejiang province. The sero-prevalence rates of R. typhi, B. henselae, R. hainan and C. burnetii were higher than that from other pathogens.

  11. The role of callous/unemotional traits in mediating the association between animal abuse exposure and behavior problems among children exposed to intimate partner violence.

    PubMed

    McDonald, Shelby Elaine; Dmitrieva, Julia; Shin, Sunny; Hitti, Stephanie A; Graham-Bermann, Sandra A; Ascione, Frank R; Williams, James Herbert

    2017-10-01

    Children exposed to intimate partner violence are at increased risk for concomitant exposure to maltreatment of companion animals. There is emerging evidence that childhood exposure to maltreatment of companion animals is associated with psychopathology in childhood and adulthood. However, few studies have explored developmental factors that might help to explain pathways from animal maltreatment exposure to children's maladjustment. The present study addresses this gap in the literature by examining relations between children's exposure to animal maltreatment, callous/unemotional traits (i.e., callousness, uncaring traits, and unemotional traits), and externalizing and internalizing behavior problems. A sample of 291 ethnically diverse children (55% Latino or Hispanic) between the ages of 7 and 12 was recruited from community-based domestic violence services. A meditational path model indicated that child exposure to animal maltreatment was associated with callousness (β=0.14), which in turn was associated with greater internalizing (β=0.32) and externalizing problems (β=0.47). The effect of animal maltreatment exposure on externalizing problems was mediated through callousness. Results suggest that callous/unemotional traits are a potential mechanism through which childhood exposure to animal maltreatment influences subsequent behavior problems. Future research is needed to evaluate the extent to which exposure to animal maltreatment affects children's adjustment over time in the context of other co-occurring adverse childhood experiences. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Detection of Toxoplasma gondii DNA in Brazilian oysters (Crassostrea rhizophorae).

    PubMed

    Ribeiro, L A; Santos, L K N S S; Brito, P A; Maciel, B M; Da Silva, A V; Albuquerque, G R

    2015-05-04

    The aim of this study was to detect evidence of Toxoplasma gondii using polymerase chain reaction (PCR)-based techniques in oysters (Crassostrea rhizophorae) obtained from the southern coastal region of Bahia, Brazil. A total of 624 oysters were collected, and the gills and digestive glands were dissected. Each tissue sample was separated into pools containing tissues (of the same type) from three animals, leading to a total of 416 experimental samples for analysis (208 samples each from the gills and digestive glands). Molecular analysis using PCR-based detection of the T. gondii AF 146527 repetitive fragment yielded negative results for all samples. However, when nested-PCR was used for detection of the T. gondii SAG-1 gene, 17 samples were positive, with the gills being the tissue with maximal detection of the parasite. These positive results were confirmed by sample sequencing. It is therefore suggested that C. rhizophorae oysters are capable of filtering and retaining T. gondii oocysts in their tissue. This represents a risk to public health because they are traditionally ingested in natura.

  13. [Detection of Cryptospordium spp. in environmental water samples by FTA-PCR].

    PubMed

    Zhang, Xiao-Ping; Zhu, Qian; He, Yan-Yan; Jiang, Li; Jiang, Shou-Fu

    2011-02-01

    To establish a FTA-polymeras chain reaction (FTA-PCR) method in detection of Cryptospordium spp. in different sources of water. The semi automated immunomagnetic separation (IMS) of Cryptospordium oocysts in environmental water samples was performed firstly, and then genomic DNA of Cryptospordium oocysts was extracted by FTA filters disk. Oligonucleotide primers were designed based on the DNA fragment of the 18 S rRNA gene from C. parvum. Plate DNA was amplified with primers in PCR. The control DNA samples from Toxoplasma gondii,Sarcocystis suihominis, Echinococcus granulosus, and Clonorchis sinensis were amplified simultaneously. All PCR products were detected by agar electrophoresis dyed with ethidium bromide. The 446 bp fragment of DNA was detected in all samples of C. parvum, C. andersoni, and C. baileyi, while it was not detected in control groups in laboratory. No positive samples were found from 10 samples collected from tape water in 5 districts of Shanghai City by FTA-PCR. Nine positive samples were detected totally from 70 different environmental water samples, there were 0 out of 15 samples from the source of tape water, 2 out of 25 from the Huangpu River, 5 out of 15 from rivers around the animal farmers, 1 out of 9 from output water of contaminating water treatment factory, 1 out of 6 from the out gate of living contaminating water. The 446 bp fragment was detected from all the amplified positive water samples. FTA-PCR is an efficient method for gene detection of Cryptospordium oocysts, which could be used in detection of environmental water samples. The contamination degree of Cryptospordium oocysts in the river water around animal farms is high.

  14. Temporal and Regional Variability in the Skin Microbiome of Humpback Whales along the Western Antarctic Peninsula.

    PubMed

    Bierlich, K C; Miller, Carolyn; DeForce, Emelia; Friedlaender, Ari S; Johnston, David W; Apprill, Amy

    2018-03-01

    The skin is the first line of defense between an animal and its environment, and disruptions in skin-associated microorganisms can be linked to an animal's health and nutritional state. To better understand the skin microbiome of large whales, high-throughput sequencing of partial small subunit rRNA genes was used to study the skin-associated bacteria of 89 seemingly healthy humpback whales ( Megaptera novaeangliae ) sampled along the Western Antarctic Peninsula (WAP) during early (2010) and late (2013) austral summers. Six core groups of bacteria were present in 93% or more of all humpback skin samples. A shift was observed in the average relative abundances of these core bacteria over time, with the emergence of four additional core groups of bacteria that corresponded to a decrease in water temperature, possibly caused by season- or foraging-related changes in skin biochemistry that influenced microbial growth, or other temporal factors. The skin microbiome differed between whales sampled at several regional locations along the WAP, suggesting that environmental factors or population may also influence the whale skin microbiome. Overall, the skin microbiome of humpback whales appears to provide insight into animal- and environment-related factors and may serve as a useful indicator for animal health or ecosystem alterations. IMPORTANCE The microbiomes of wild animals are currently understudied but may provide information about animal health and/or animal-environment interactions. In the largest sampling of any marine mammal microbiome, this study demonstrates conservation in the skin microbiome of 89 seemingly healthy humpback whales sampled in the Western Antarctic Peninsula, with shifts in the bacterial community composition related to temporal and regional variability. This study is important because it suggests that the skin microbiome of humpback whales could provide insight into animal nutritional or seasonal/environment-related factors, which are becoming increasingly important to recognize due to unprecedented rates of climate change and anthropogenic impact on ocean ecosystems. Copyright © 2018 Bierlich et al.

  15. Temporal and Regional Variability in the Skin Microbiome of Humpback Whales along the Western Antarctic Peninsula

    PubMed Central

    Bierlich, K. C.; Miller, Carolyn; DeForce, Emelia; Friedlaender, Ari S.

    2017-01-01

    ABSTRACT The skin is the first line of defense between an animal and its environment, and disruptions in skin-associated microorganisms can be linked to an animal's health and nutritional state. To better understand the skin microbiome of large whales, high-throughput sequencing of partial small subunit rRNA genes was used to study the skin-associated bacteria of 89 seemingly healthy humpback whales (Megaptera novaeangliae) sampled along the Western Antarctic Peninsula (WAP) during early (2010) and late (2013) austral summers. Six core groups of bacteria were present in 93% or more of all humpback skin samples. A shift was observed in the average relative abundances of these core bacteria over time, with the emergence of four additional core groups of bacteria that corresponded to a decrease in water temperature, possibly caused by season- or foraging-related changes in skin biochemistry that influenced microbial growth, or other temporal factors. The skin microbiome differed between whales sampled at several regional locations along the WAP, suggesting that environmental factors or population may also influence the whale skin microbiome. Overall, the skin microbiome of humpback whales appears to provide insight into animal- and environment-related factors and may serve as a useful indicator for animal health or ecosystem alterations. IMPORTANCE The microbiomes of wild animals are currently understudied but may provide information about animal health and/or animal-environment interactions. In the largest sampling of any marine mammal microbiome, this study demonstrates conservation in the skin microbiome of 89 seemingly healthy humpback whales sampled in the Western Antarctic Peninsula, with shifts in the bacterial community composition related to temporal and regional variability. This study is important because it suggests that the skin microbiome of humpback whales could provide insight into animal nutritional or seasonal/environment-related factors, which are becoming increasingly important to recognize due to unprecedented rates of climate change and anthropogenic impact on ocean ecosystems. PMID:29269499

  16. Identification of cattle, buffaloes and rodents as reservoir animals of Leptospira in the District of Gampaha, Sri Lanka.

    PubMed

    Denipitiya, D T H; Chandrasekharan, N V; Abeyewickreme, W; Hartskeerl, R A; Hapugoda, M D

    2017-03-23

    Leptospirosis is an important emerging infectious disease in Sri Lanka. Rats are the most important reservoir of Leptospira but domestic and wild mammals may also act as important maintenance or accidental hosts. In Sri Lanka, knowledge of reservoir animals of leptospires is poor. The objective of this study was to identify potential reservoir animals of Leptospira in the District of Gampaha, Sri Lanka. Blood and kidney samples were collected from 38 rodents and mid-stream urine samples were randomly collected from 45 cattle and five buffaloes in the District of Gampaha. Kidney and urine samples were tested by real-time polymerase chain reaction (PCR) and serum samples were tested by the microscopic agglutination test (MAT). Of the 38 rodent kidney samples, 11% (4/38) were positive by real-time PCR. The prevalence of leptospiral carriage was 11% (3/26) and 8% (1/12) in female and male rodents, respectively. Three rodent serum samples were positive by MAT. Of the 50 cattle/buffalo urine samples tested, 10% (5/50) were positive by real-time PCR. The prevalence of leptospiral carriage was 9% (4/45) and 20% (1/5) in cattle and buffaloes, respectively. Results of PCR and MAT showed that Leptospira were present in a significant proportion of the rodents and farm animals tested in this study and suggest that these (semi-) domestic animals form an infection reservoir for Leptospira. Therefore, there is a potential zoonotic risk to public health, most notably to farmers in this area.

  17. [Molecular typing of 12 Brucella strains isolated in Guizhou province in 2010-2013].

    PubMed

    Wang, Yue; Chen, Hong; Liu, Ying; Zhou, Jingzhu; Li, Shijun; Hang, Yan; Tang, Guangpeng; Wang, Dingming; Chen, Guichun

    2015-09-01

    To identify and characterize the Brucella strains from Guizhou province in 2010-2013. A total of 12 strains of Brucella suspicious bacteria were isolated in Guizhou province from 2010 to 2013. Four strains (GZLL3, GZLL4, GZLL11 and SH2) were isolated from goat blood samples and eight strains (SH4, GZZY, GZSQ, GZZA, BR13001, BR13004, BR13005 and BR13006) were isolated from blood samples of patient 12 Brucella suspicious strains were identified and characterized using conventional methods. Brucella genus specific gene BCSP31-based PCR (BCSP31-PCR) was used to identify the genus of Brucella and IS711 insert sequence-based PCR (AMOS-PCR) was applied to identify the species of Brucella strains. Goats and patients originated Brucella strains were comparatively analysed using Pulse-field Gel Electrophoresis (PFGE). Both of conventional methods and PCR identified the 12 Brucella suspicious strains as B. melitensis biotype 3. BCSP31-PCR identification results showed that a specific DNA bands (223 bp) were detected in all the 12 strains and positive control samples with no DNA band in negative samples. AMOS-PCR amplified a 731 bp-DNA bands in all the 12 strains, with 731 bp, 498 bp and 275 bp in M5, S2 and A19 strains, respectively, and no DNA band was detected in the negative control samples. PFGE analysis showed that 12 Brucella isolates from patients and goats showed consistent PFGE patterns with the digestion of restriction enzyme Xba I. The epidemic species/type of Brucella in both human and animal in Guizhou province was B. melitensis biotype 3 and goat was the main animal source of infection of brucellosis in Guizhou province.

  18. Quantitative CrAssphage PCR Assays for Human Fecal ...

    EPA Pesticide Factsheets

    Environmental waters are monitored for fecal pollution to protect public health and water resources. Traditionally, general fecal indicator bacteria are used; however, they cannot distinguish human fecal waste from pollution from other animals. Recently, a novel bacteriophage, crAssphage, was discovered by metagenomic data mining and reported to be abundant in and closely associated with human fecal waste. To confirm bioinformatic predictions, 384 primer sets were designed along the length of the crAssphage genome. Based upon initial screening, two novel crAssphage qPCR assays (CPQ_056 and CPQ_064) were designed and evaluated in reference fecal samples and water matrices. The assays exhibited high specificities (98.6%) when tested against a large animal fecal reference library and were highly abundant in raw sewage and sewage impacted water samples. In addition, CPQ_056 and CPQ_064 assay performance was compared to HF183/BacR287 and HumM2 methods in paired experiments. Findings confirm viral crAssphage qPCR assays perform at a similar level to well established bacterial human-associated fecal source identification technologies. These new viral based assays could become important water quality management and research tools. To inform the public.

  19. In vivo degradation of polyurethane foam with 55 wt % polyethylene glycol.

    PubMed

    Broekema, Ferdinand I; Van Leeuwen, M Barbara M; Van Minnen, Baucke; Bos, Rudolf R M

    2015-11-01

    Most topical hemostatic agents are based on animal-derived products like collagen and gelatin. They carry the potential risk of pathogen transmission while adjustments in the production process of these materials are limited. A synthetic hemostatic agent based on polyurethane (PU) and polyethylene glycol (PEG) was developed to overcome these disadvantages. The goal of this study was to compare the degradation process of this biomaterial to collagen and gelatin hemostatic agents. Samples of the test materials were implanted subcutaneously in both rats and rabbits. The animals were sacrificed at certain time intervals up to three years and the explanted samples were microscopically assessed. The histological examination showed a comparable pattern of degradation for the different test materials. Remnants of gelatin and collagen were seen up to 26 and 39 weeks, respectively. For PU, it took up to three years before micro-particles of the material were no longer detected. All biomaterials showed a good biocompatibility and no severe foreign body reactions occurred. The good biocompatibility and predictable pattern of resorption indicate that PU can be used as a topical hemostatic agent. However, a degradation time comparable to collagen and gelatin would be favorable. © 2015 Wiley Periodicals, Inc.

  20. The detection and recovery of Erysipelothrix spp. in meat and abattoir samples in Western Australia.

    PubMed

    Wang, Q; Fidalgo, S; Chang, B J; Mee, B J; Riley, T V

    2002-01-01

    To investigate the occurrence of Erysipelothrix rhusiopathiae and other Erysipelothrix spp. in abattoir and meat samples in Western Australia. Samples were collected from various parts of pig and sheep carcasses, as well as different sections of slaughtering line, pen soil and effluent. Previously evaluated culture methods were applied for the isolation of Erysipelothrix spp., in conjunction with phenotypic and genotypic detection and identification procedures. Of 109 samples from the two abattoirs, 35 (32.1%) were Erysipelothrix genus-specific PCR-positive. These came from swabs of animal exterior surfaces and joints, slaughtering areas, pig pen soil and abattoir effluent. Four samples (3.7%) from sheep arthritic joints and pig abattoir effluent were also E. rhusiopathiae species-specific PCR-positive. Of 123 carcass washing samples, 12 (9.8%) were genus-specific PCR-positive, and these came from all five kinds of meat samples tested, including beef, lamb, mutton, pork and chicken. Four of them (3.3%) were also species-specific PCR-positive. A total of 25 isolates was recovered from the samples, of which seven were identified as E. rhusiopathiae, seven were consistent with E. tonsillarum, and the remaining 11 were other species of Erysipelothrix. Erysipelothrix spp. can still be isolated and identified from specimens of animal origin with relative ease, provided that appropriate cultural and molecular procedures are used. Clinical microbiology laboratories may need to improve their diagnostic protocols. This study confirms that E. rhusiopathiae and other species of Erysipelothrix continue to colonize and contaminate farmed animals and animal products. Erysipelothrix infection still poses a potential threat to the economy of the farmed animal industry, as well as being a potential human public health hazard.

  1. Health assessment of free-living eastern box turtles (Terrapene carolina carolina) in and around the Maryland Zoo in Baltimore 1996-2011.

    PubMed

    Adamovicz, Laura; Bronson, Ellen; Barrett, Kevin; Deem, Sharon L

    2015-03-01

    Health data for free-living eastern box turtles (Terrapene carolina carolina) at the Maryland Zoo in Baltimore were analyzed. One hundred and eighteen turtles were captured on or near zoo grounds over the course of 15 yr (1996-2011), with recapture of many individuals leading to 208 total evaluations. Of the 118 individuals, 61 were male, 50 were female, and 7 were of undetermined sex. Of the 208 captures, 188 were healthy, and 20 were sick or injured. Complete health evaluations were performed on 30 turtles with physical examination records, complete blood counts (CBCs), and plasma biochemistry profiles. Eight animals were sampled more than once, yielding 40 total samples for complete health evaluations of these 30 individuals. The 40 samples were divided into healthy (N=29) and sick (N=11) groups based on clinical findings on physical examination. Samples from healthy animals were further divided into male (N=17) and female (N= 12) groups. CBC and biochemistry profile parameters were compared between sick and healthy groups and between healthy males and females. Sick turtles had lower albumin, globulin, total protein (TP), calcium, phosphorous, sodium, and potassium than healthy animals. Sick turtles also had higher heterophil to lymphocyte ratios. Healthy female turtles had higher leukocyte count, eosinophil count, total solids, TP, globulin, cholesterol, calcium, and phosphorous than healthy males. Banked plasma from all 40 samples was tested for antibodies to Mycoplasma agassizii and Mycoplasma testudineum via enzyme-linked immunosorbent assay. One sample from a clinically healthy female was antibody positive for M. agassizii; none were positive for M. testudineum. This study provides descriptive health data for eastern box turtles and CBC and biochemistry profile information for T. carolina carolina at and near the Maryland Zoo in Baltimore. It also reports low serologic evidence of exposure to mycoplasmosis.

  2. Survey of Malassezia sp and dermatophytes in the cutaneous microbiome of free-ranging golden-headed lion tamarins (Leontopithecus chrysomelas - Kuhl, 1820).

    PubMed

    Neves, Juan Ja; Francelino, Marcelo; Silva, Flavia Gl; Baptista, Luana Cl; Bueno, Marina G; Catão-Dias, José L; Molina, Camila; Kierulff, Maria Cm; Pissinatti, Alcides; Coutinho, Selene DA

    2017-06-01

    Data about the presence of fungi on the cutaneous surface of wild animals are scarce. The aim of this study was to survey dermatophytes and Malassezia sp in the external ear canal and haircoat of Leontopithecus chrysomelas. A total of 928 clinical samples were collected from 232 animals: For Malassezia screening 696 samples were studied, 464 of cerumen and 232 of haircoat; another 232 haircoat samples were studied for dermatophyte analysis. A geophilic dermatophyte, Microsporum cookie, was isolated from one young female. Lipodependent Malassezia was isolated from 76 animals and 87 clinical samples, 26 from the cerumen and 61 from the haircoat (statistically significant); there were no differences related to gender and age. Results suggested that lipodependent Malassezia is part of the skin microbiome of these animals. The prevalence of dermatophytes was too low and probably not relevant for the health of the studied population. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  3. Effects of a fire alarm strobe light on fecal corticosterone metabolite concentrations in mice.

    PubMed

    Godfrey, Denice; Silverman, Jerald

    2009-02-01

    The type and location of fire alarms are important considerations in animal facility design. The Guide for the Care and Use of Laboratory Animals recommends minimizing animal exposure to such alarms. Nevertheless, it is often necessary to maintain fire alarms within animal housing or procedural areas. The authors exposed male mice to the flashing strobe light component of a standard fire alarm and evaluated mouse fecal corticosterone concentration, which is known to be an indicator of stress. Mice were exposed to the strobe light for 5 min during either the light or the dark phase of the light:dark cycle. The authors collected fecal samples every 6 h for 24 h before exposing mice to the alarm and every 6 h for 24 h after exposure. Fecal samples taken before exposure (baseline samples) showed a normal circadian pattern of corticosterone metabolite excretion. In fecal samples taken after mice were exposed to the fire alarm, metabolite concentrations did not significantly differ from baseline concentrations over time.

  4. Rickettsia spp. among wild mammals and their respective ectoparasites in Pantanal wetland, Brazil.

    PubMed

    de Sousa, Keyla Carstens Marques; Herrera, Heitor Miraglia; Rocha, Fabiana Lopes; Costa, Francisco Borges; Martins, Thiago Fernandes; Labruna, Marcelo Bahia; Machado, Rosangela Zacarias; André, Marcos Rogério

    2018-01-01

    The genus Rickettsia comprises obligatory intracellular bacteria, well known to cause zoonotic diseases around the world. The present work aimed to investigate the occurrence of Rickettsia spp. in wild animals, domestic dogs and their respective ectoparasites in southern Pantanal region, central-western Brazil, by molecular and serological techniques. Between August 2013 and March 2015, serum, whole blood and/or spleen samples were collected from 31 coatis, 78 crab-eating foxes, seven ocelots, 42 dogs, 110 wild rodents, and 30 marsupials. Serum samples from canids, felids, rodents and marsupials were individually tested by indirect fluorescent antibody test (IFAT) in order to detect IgG antibodies to Rickettsia rickettsii, Rickettsia parkeri and Rickettsia amblyommatis. DNA samples from mammals and ectoparasites were submitted to a multiplex qPCR assay in order to detect and quantify spotted fever group (SFG) and typhus group (TG) rickettsiae and Orientia tsutsugamushi. Positive samples in qPCR assays were submitted to conventional PCR assays targeting gltA, ompA, ompB and htrA genes, followed by sequencing and phylogenetic analyses. The ticks collected (1582) from animals belonged to the species Amblyomma sculptum, Amblyomma parvum, Amblyomma ovale, Amblyomma tigrinum, Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus sensu lato and Amblyomma auricularium. Overall, 27 (64.2%) dogs, 59 (75.6%) crab-eating foxes and six (85.7%) ocelots were seroreactive (titer≥64) to at least one Rickettsia species. For 17 (40.4%) dogs, 33 (42.3%) crab-eating foxes, and two (33.3%) ocelots, homologous reactions to R. amblyommatis or a closely related organism were suggested. One hundred and sixteen (23.5%) tick samples and one (1.2%) crab-eating fox blood sample showed positivity in qPCR assays for SFG Rickettsia spp. Among SFG Rickettsia-positive ticks samples, 93 (80.2%) belonged to A. parvum, 14 (12%) belonged to A. sculptum species, three (2.5%) belonged to A. auricularim, and six (5.2%) were Amblyomma larval pools. Thirty samples out of 117 qPCR positive samples for SFG Rickettsia spp. also showed positivity in cPCR assays based on gltA, htrA and/or ompB genes. The Blast analyses showed 100% identity with 'Candidatus Rickettsia andeanae' in all 30 sequences obtained from gltA, htrA and/or ompB genes. The concatenated phylogenetic analysis based on gltA and 17-kDa htrA genes grouped the Rickettsia sequences obtained from tick samples in the same clade of 'Candidatus Rickettsia andeanae'. The present study revealed that wild and domestic animals in southern Pantanal region, Brazil, are exposed to SFG rickettsiae agents. Future studies regarding the pathogenicity of these agents are necessary in order to prevent human cases of rickettsiosis in Brazilian southern Pantanal. Copyright © 2017 Elsevier GmbH. All rights reserved.

  5. Understanding Giardia infections among rural communities using the one health approach.

    PubMed

    Lee, Soo Ching; Ngui, Romano; Tan, Tiong Kai; Roslan, Muhammad Aidil; Ithoi, Init; Mahdy, Mohammed A K; Hakim S, Lokman; Lim, Yvonne A L

    2017-12-01

    The epidemiology of giardiasis in rural villages in Peninsular Malaysia was examined in the context of the One Health triad that encompasses humans, animals and environment (i.e. river water). A cross-sectional study was carried out among five rural communities in Malaysia to determine the prevalence of Giardia duodenalis in humans, animals and river water. Fecal samples collected from humans and animals were examined by light microscopy. Water was sampled from the rivers adjacent to the target communities and investigated for the occurrence of Giardia cysts. The isolated cysts were further genotyped targeting the glutamate dehydrogenase and triosephosphate isomerase genes. The overall prevalence of G. duodenalis was 6.7% (18/269) and 4.7% (8/169) among humans and animals, respectively. Giardia cysts (mean concentration range: 0.10-5.97 cysts/L) were also found in adjacent rivers at four out of the five villages examined. At Kemensah and Kuala Pangsun, Giardia cysts were isolated from humans [rate: 3.7% each (of 54 each)], animals [rates: 6.3% (of 62) and 11.3% (of 16), respectively] and river water [average concentration of 9 samples each: 0.83±0.81 and 5.97±7.00, respectively]. For both villages at Pos Piah and Paya Lebar, 12.2% (of 98) and 6.1% (of 33) of collected human samples were infected, respectively whilst none of the collected animals samples in these villages were found to be positive. The river water samples of these two villages were also contaminated (average concentration: 0.20±0.35 (of 9) and 0.10±0.19 (of 3), respectively). In conclusion, Giardia cysts were simultaneously observed in the human-animal-environment (i.e., river water) interfaces in at least two of five studied communities highlighting a vital need to improve understanding on the interplay of transmission dynamics, the role of infected humans and animals in contaminating the water sources and the role of water as a vehicle of disease transmission in these communities. Indeed, this study illustrates the One Health approach which is to recognize that the optimal health of humans are interconnected with the well-being of animals and their environment. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Multi-mycotoxin analysis of animal feed and animal-derived food using LC-MS/MS system with timed and highly selective reaction monitoring.

    PubMed

    Zhao, Zhiyong; Liu, Na; Yang, Lingchen; Deng, Yifeng; Wang, Jianhua; Song, Suquan; Lin, Shanhai; Wu, Aibo; Zhou, Zhenlei; Hou, Jiafa

    2015-09-01

    Mycotoxins have the potential to enter the human food chain through carry-over of contaminants from feed into animal-derived products. The objective of the study was to develop a reliable and sensitive method for the analysis of 30 mycotoxins in animal feed and animal-derived food (meat, edible animal tissues, and milk) using liquid chromatography-tandem mass spectrometry (LC-MS/MS). In the study, three extraction procedures, as well as various cleanup procedures, were evaluated to select the most suitable sample preparation procedure for different sample matrices. In addition, timed and highly selective reaction monitoring on LC-MS/MS was used to filter out isobaric matrix interferences. The performance characteristics (linearity, sensitivity, recovery, precision, and specificity) of the method were determined according to Commission Decision 2002/657/EC and 401/2006/EC. The established method was successfully applied to screening of mycotoxins in animal feed and animal-derived food. The results indicated that mycotoxin contamination in feed directly influenced the presence of mycotoxin in animal-derived food. Graphical abstract Multi-mycotoxin analysis of animal feed and animal-derived food using LC-MS/MS.

  7. Low-dose 4D cardiac imaging in small animals using dual source micro-CT

    NASA Astrophysics Data System (ADS)

    Holbrook, M.; Clark, D. P.; Badea, C. T.

    2018-01-01

    Micro-CT is widely used in preclinical studies, generating substantial interest in extending its capabilities in functional imaging applications such as blood perfusion and cardiac function. However, imaging cardiac structure and function in mice is challenging due to their small size and rapid heart rate. To overcome these challenges, we propose and compare improvements on two strategies for cardiac gating in dual-source, preclinical micro-CT: fast prospective gating (PG) and uncorrelated retrospective gating (RG). These sampling strategies combined with a sophisticated iterative image reconstruction algorithm provide faster acquisitions and high image quality in low-dose 4D (i.e. 3D  +  Time) cardiac micro-CT. Fast PG is performed under continuous subject rotation which results in interleaved projection angles between cardiac phases. Thus, fast PG provides a well-sampled temporal average image for use as a prior in iterative reconstruction. Uncorrelated RG incorporates random delays during sampling to prevent correlations between heart rate and sampling rate. We have performed both simulations and animal studies to validate these new sampling protocols. Sampling times for 1000 projections using fast PG and RG were 2 and 3 min, respectively, and the total dose was 170 mGy each. Reconstructions were performed using a 4D iterative reconstruction technique based on the split Bregman method. To examine undersampling robustness, subsets of 500 and 250 projections were also used for reconstruction. Both sampling strategies in conjunction with our iterative reconstruction method are capable of resolving cardiac phases and provide high image quality. In general, for equal numbers of projections, fast PG shows fewer errors than RG and is more robust to undersampling. Our results indicate that only 1000-projection based reconstruction with fast PG satisfies a 5% error criterion in left ventricular volume estimation. These methods promise low-dose imaging with a wide range of preclinical applications in cardiac imaging.

  8. Ultra-low-density genotype panels for breed assignment of Angus and Hereford cattle.

    PubMed

    Judge, M M; Kelleher, M M; Kearney, J F; Sleator, R D; Berry, D P

    2017-06-01

    Angus and Hereford beef is marketed internationally for apparent superior meat quality attributes; DNA-based breed authenticity could be a useful instrument to ensure consumer confidence on premium meat products. The objective of this study was to develop an ultra-low-density genotype panel to accurately quantify the Angus and Hereford breed proportion in biological samples. Medium-density genotypes (13 306 single nucleotide polymorphisms (SNPs)) were available on 54 703 commercial and 4042 purebred animals. The breed proportion of the commercial animals was generated from the medium-density genotypes and this estimate was regarded as the gold-standard breed composition. Ten genotype panels (100 to 1000 SNPs) were developed from the medium-density genotypes; five methods were used to identify the most informative SNPs and these included the Delta statistic, the fixation (F st) statistic and an index of both. Breed assignment analyses were undertaken for each breed, panel density and SNP selection method separately with a programme to infer population structure using the entire 13 306 SNP panel (representing the gold-standard measure). Breed assignment was undertaken for all commercial animals (n=54 703), animals deemed to contain some proportion of Angus based on pedigree (n=5740) and animals deemed to contain some proportion of Hereford based on pedigree (n=5187). The predicted breed proportion of all animals from the lower density panels was then compared with the gold-standard breed prediction. Panel density, SNP selection method and breed all had a significant effect on the correlation of predicted and actual breed proportion. Regardless of breed, the Index method of SNP selection numerically (but not significantly) outperformed all other selection methods in accuracy (i.e. correlation and root mean square of prediction) when panel density was ⩾300 SNPs. The correlation between actual and predicted breed proportion increased as panel density increased. Using 300 SNPs (selected using the global index method), the correlation between predicted and actual breed proportion was 0.993 and 0.995 in the Angus and Hereford validation populations, respectively. When SNP panels optimised for breed prediction in one population were used to predict the breed proportion of a separate population, the correlation between predicted and actual breed proportion was 0.034 and 0.044 weaker in the Hereford and Angus populations, respectively (using the 300 SNP panel). It is necessary to include at least 300 to 400 SNPs (per breed) on genotype panels to accurately predict breed proportion from biological samples.

  9. 9 CFR 113.29 - Determination of moisture content in desiccated biological products.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Determination of moisture content in desiccated biological products. 113.29 Section 113.29 Animals and Animal Products ANIMAL AND PLANT HEALTH... as “A.” (iii) Return weighing bottles to the desiccator. (2) Remove the sample container seal. (i...

  10. 9 CFR 113.29 - Determination of moisture content in desiccated biological products.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Determination of moisture content in desiccated biological products. 113.29 Section 113.29 Animals and Animal Products ANIMAL AND PLANT HEALTH... as “A.” (iii) Return weighing bottles to the desiccator. (2) Remove the sample container seal. (i...

  11. Development of a Blocking ELISA Using a Monoclonal Antibody to a Dominant Epitope in Non-Structural Protein 3A of Foot-and-Mouth Disease Virus, as a Matching Test for a Negative-Marker Vaccine.

    PubMed

    Fu, Yuanfang; Li, Pinghua; Cao, Yimei; Wang, Na; Sun, Pu; Shi, Qian; Ji, Xincheng; Bao, Huifang; Li, Dong; Chen, Yingli; Bai, Xingwen; Ma, Xueqing; Zhang, Jing; Lu, Zengjun; Liu, Zaixin

    2017-01-01

    Foot-and-mouth disease (FMD) is a devastating animal disease. Strategies for differentiation of infected from vaccinated animals (DIVA) remain very important for controlling disease. Development of an epitope-deleted marker vaccine and accompanying diagnostic method will improve the efficiency of DIVA. Here, a monoclonal antibody (Mab) was found to recognize a conserved "AEKNPLE" epitope spanning amino acids 109-115 of non-structural protein (NSP) 3A of foot-and-mouth disease virus (FMDV; O/Tibet/CHA/99 strain), which could be deleted by a reverse-genetic procedure. In addition, a blocking ELISA was developed based on this Mab against NSP 3A, which could serve as a matching test for a negative-marker vaccine. The criterion of this blocking ELISA was determined by detecting panels of sera from different origins. The serum samples with a percentage inhibition (PI) equal or greater than 50% were considered to be from infected animals, and those with <50% PI were considered to be from non-infected animals. This test showed similar performance when compared with other 2 blocking ELISAs based on an anti-NSP 3B Mab. This is the first report of the DIVA test for an NSP antibody based on an Mab against the conserved and predominant "AEKNPLE" epitope in NSP 3A of FMDV.

  12. A High-Throughput Method for Direct Detection of Therapeutic Oligonucleotide-Induced Gene Silencing In Vivo

    PubMed Central

    Coles, Andrew H.; Osborn, Maire F.; Alterman, Julia F.; Turanov, Anton A.; Godinho, Bruno M.D.C.; Kennington, Lori; Chase, Kathryn; Aronin, Neil

    2016-01-01

    Preclinical development of RNA interference (RNAi)-based therapeutics requires a rapid, accurate, and robust method of simultaneously quantifying mRNA knockdown in hundreds of samples. The most well-established method to achieve this is quantitative real-time polymerase chain reaction (qRT-PCR), a labor-intensive methodology that requires sample purification, which increases the potential to introduce additional bias. Here, we describe that the QuantiGene® branched DNA (bDNA) assay linked to a 96-well Qiagen TissueLyser II is a quick and reproducible alternative to qRT-PCR for quantitative analysis of mRNA expression in vivo directly from tissue biopsies. The bDNA assay is a high-throughput, plate-based, luminescence technique, capable of directly measuring mRNA levels from tissue lysates derived from various biological samples. We have performed a systematic evaluation of this technique for in vivo detection of RNAi-based silencing. We show that similar quality data is obtained from purified RNA and tissue lysates. In general, we observe low intra- and inter-animal variability (around 10% for control samples), and high intermediate precision. This allows minimization of sample size for evaluation of oligonucleotide efficacy in vivo. PMID:26595721

  13. On the choice of statistical models for estimating occurrence and extinction from animal surveys

    USGS Publications Warehouse

    Dorazio, R.M.

    2007-01-01

    In surveys of natural animal populations the number of animals that are present and available to be detected at a sample location is often low, resulting in few or no detections. Low detection frequencies are especially common in surveys of imperiled species; however, the choice of sampling method and protocol also may influence the size of the population that is vulnerable to detection. In these circumstances, probabilities of animal occurrence and extinction will generally be estimated more accurately if the models used in data analysis account for differences in abundance among sample locations and for the dependence between site-specific abundance and detection. Simulation experiments are used to illustrate conditions wherein these types of models can be expected to outperform alternative estimators of population site occupancy and extinction. ?? 2007 by the Ecological Society of America.

  14. Microbiota and anthropic interference on antimicrobial resistance profile of bacteria isolated from Brazilian Maned-wolf (Chrysocyon brachyurus).

    PubMed

    Vieira-da-Motta, Olney; Eckhardt-de-Pontes, Luiz Antonio; Petrucci, Melissa Paes; dos Santos, Israel Pereira; da Cunha, Isabel Candia Nunes; Morato, Ronaldo Gonçalves

    2013-12-01

    Both the study of Brazilian wild mammal fauna and the conditions that foster the preservation of endangered species, such as Brazilian Maned-wolf (Chrysocyon brachyurus), in wild life are of extreme importance. In order to study the resistance profile of microbiota bacterial colonizing Brazilian Maned-wolf, this work investigated samples from eight male captive and free roaming animals originating from different Brazilian geographical regions. Samples for microbiological purposes were collected with swabs and kept in appropriate transport medium. Using routine microbiological techniques, the isolated bacteria were tested toward antimicrobial drugs by the agar disk diffusion method. Results showed that all samples from wild animals were sensitive toward all drugs tested. Conversely, the resistance profile of bacteria isolated from captive animals varied among strains and animal body site location. Escherichia coli samples from prepuce, anus and ear showed multi-resistance toward at least four drugs, especially against erythromycin and tetracycline, followed by Proteus mirabilis and P. vulgaris strains isolated from anus and ear. Among Gram-positive bacteria, strains of coagulase-negative staphylococci showed multi-resistance mainly toward erythromycin and amoxicillin. The work discusses these findings and suggests that profile of multi-resistance bacteria from captive subjects may be attributed to direct contact with human or through lifestyle factors such as feeding, predation or contact of animals with urban animals such as birds, rodents, and insects from surrounding environments.

  15. Microbiota and anthropic interference on antimicrobial resistance profile of bacteria isolated from Brazilian maned-wolf (Chrysocyon brachyurus)

    PubMed Central

    Vieira-da-Motta, Olney; Eckhardt-de-Pontes, Luiz Antonio; Petrucci, Melissa Paes; dos Santos, Israel Pereira; da Cunha, Isabel Candia Nunes; Morato, Ronaldo Gonçalves

    2013-01-01

    Both the study of Brazilian wild mammal fauna and the conditions that foster the preservation of endangered species, such as Brazilian Maned-wolf (Chrysocyon brachyurus), in wild life are of extreme importance. In order to study the resistance profile of microbiota bacterial colonizing Brazilian Maned-wolf, this work investigated samples from eight male captive and free roaming animals originating from different Brazilian geographical regions. Samples for microbiological purposes were collected with swabs and kept in appropriate transport medium. Using routine microbiological techniques, the isolated bacteria were tested toward antimicrobial drugs by the agar disk diffusion method. Results showed that all samples from wild animals were sensitive toward all drugs tested. Conversely, the resistance profile of bacteria isolated from captive animals varied among strains and animal body site location. Escherichia coli samples from prepuce, anus and ear showed multi-resistance toward at least four drugs, especially against erythromycin and tetracycline, followed by Proteus mirabilis and P. vulgaris strains isolated from anus and ear. Among Gram-positive bacteria, strains of coagulase-negative staphylococci showed multi-resistance mainly toward erythromycin and amoxicillin. The work discusses these findings and suggests that profile of multi-resistance bacteria from captive subjects may be attributed to direct contact with human or through lifestyle factors such as feeding, predation or contact of animals with urban animals such as birds, rodents, and insects from surrounding environments. PMID:24688529

  16. Abattoir based surveillance of cystic echinococcosis (CE) in the Sultanate of Oman during 2010-2013.

    PubMed

    Al Kitani, Fadya Abdullah; Al Riyami, Shumoos; Al Yahyai, Sabra; Al Rawahi, Abdulmajeed Hamood; Al Maawali, Mahir; Al Awahi, Abdulmajeed Hamood; Al Aawali, Mahir; Hussain, Muhammad Hammad

    2015-07-30

    A passive surveillance was conducted in 8 selected abattoirs belonging to 8 governorates of Oman. Routine slaughtering of 282.020 animals was observed during 2010-2013 by duty veterinarians and suspected samples (n=1440) for cystic echinococcosis (CE) from camels (n=292), cattle (n=303), goats (n=682) and sheep (n=163) were collected. In total, 587 (40.8%) samples were found positive for CE and the highest percentage (64.2%) of positive samples belonged to Dhofar (Salalah) as compared to other governorates, p<0.01. The infection rate of CE was calculated as 5.3, 0.6, 0.07 and 0.03 in camel, cattle, sheep and goats respectively. Fertile cysts were found in 30.7% of positive samples and camels (52%) were found as major source of these followed by cattle (14.1%) and goats (9.7%). Percentage of positive to submitted samples was higher in females (67.9%) than males (29.8%). The fertility of cyst increased with age and the highest percentage of fertile cysts were collected from animals above 5 years of age (54.4%) followed by between 3 and 5 years (36.7%) and below 3 years of age (8.9%). For different breeds, 478 (62.7%) samples from local breeds of livestock were positive for CE as compared to 108 (15.9%) from imported animals (OR 8.92), p<0.01. The highest percent of positive samples was observed in cysts from lungs (54.4%) followed by those from liver (25.7%), liver and lungs (14.1%) and others (3.8%). Molecular investigations indicated that G1 and G6/7 were the two strains of CE circulating in camels and cattle while G1 was the only strain found in samples from small ruminants. Results indicated that CE is endemic in the livestock of Oman and further molecular epidemiological work in different host species is required for the conceptualization of a comprehensive control program. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Evidence of natural transmission of group A rotavirus between domestic pigs and wild boars (Sus scrofa) in Japan.

    PubMed

    Okadera, Kota; Abe, Masako; Ito, Naoto; Morikawa, Shigeki; Yamasaki, Ari; Masatani, Tatsunori; Nakagawa, Keisuke; Yamaoka, Satoko; Sugiyama, Makoto

    2013-12-01

    Group A rotaviruses (RVAs) are a major cause of acute dehydrating diarrhea in infants and young animals worldwide. RVAs have also been detected in several wild and zoo animals, indicating wide susceptibility of wild animals. However, the role of wild animals in the infection cycle of RVAs is unclear. Wild boars are indigenous in many countries in the world. Japanese wild boars (Sus scrofa leucomystax) have been migrating close to human habitats in Japan, indicating the possibility of natural transmission between domestic animals or humans and wild boars. We investigated infection of RVAs in wild boars in Japan to identify types of RVAs infecting wild animals. We obtained stool samples from 90 wild boars and detected a VP4 gene of RVAs by RT-semi-nested PCR. RVAs were detected in samples from four of the 90 wild boars. Nucleotide analyses of VP7 and VP4 genes revealed that the four strains belong to G9P[23], G4P[23], G9P[13] and G4P[6], suggesting a relation to porcine and human RVAs. We therefore characterized RVAs circulating among domestic pigs living in the same area as the wild boars. We collected stool samples from 82 domestic pigs. RVAs were detected in samples from 49 of the 82 domestic pigs. Phylogenetic and similarity analyses provided evidence for natural transmission between domestic pigs and wild boars. The results also suggested that natural reassortment events occurred before or after transmission between domestic pigs and wild boars. Our findings indicate the possibility that RVAs circulate among wild animals, humans and domestic animals in nature. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. The potential of Fourier transform infrared spectroscopy of milk samples to predict energy intake and efficiency in dairy cows.

    PubMed

    McParland, S; Berry, D P

    2016-05-01

    Knowledge of animal-level and herd-level energy intake, energy balance, and feed efficiency affect day-to-day herd management strategies; information on these traits at an individual animal level is also useful in animal breeding programs. A paucity of data (especially at the individual cow level), of feed intake in particular, hinders the inclusion of such attributes in herd management decision-support tools and breeding programs. Dairy producers have access to an individual cow milk sample at least once daily during lactation, and consequently any low-cost phenotyping strategy should consider exploiting measureable properties in this biological sample, reflecting the physiological status and performance of the cow. Infrared spectroscopy is the study of the interaction of an electromagnetic wave with matter and it is used globally to predict milk quality parameters on routinely acquired individual cow milk samples and bulk tank samples. Thus, exploiting infrared spectroscopy in next-generation phenotyping will ensure potentially rapid application globally with a negligible additional implementation cost as the infrastructure already exists. Fourier-transform infrared spectroscopy (FTIRS) analysis is already used to predict milk fat and protein concentrations, the ratio of which has been proposed as an indicator of energy balance. Milk FTIRS is also able to predict the concentration of various fatty acids in milk, the composition of which is known to change when body tissue is mobilized; that is, when the cow is in negative energy balance. Energy balance is mathematically very similar to residual energy intake (REI), a suggested measure of feed efficiency. Therefore, the prediction of energy intake, energy balance, and feed efficiency (i.e., REI) from milk FTIRS seems logical. In fact, the accuracy of predicting (i.e., correlation between predicted and actual values; root mean square error in parentheses) energy intake, energy balance, and REI from milk FTIRS in dairy cows was 0.88 (20.0MJ), 0.78 (18.6MJ), and 0.63 (22.0MJ), respectively, based on cross-validation. These studies, however, are limited to results from one research group based on data from 2 contrasting production systems in the United Kingdom and Ireland and would need to be replicated, especially in a range of production systems because the prediction equations are not accurate when the variability used in validation is not represented in the calibration data set. Heritable genetic variation exists for all predicted traits. Phenotypic differences in energy intake also exists among animals stratified based on genetic merit for energy intake predicted from milk FTIRS, substantiating the usefulness of such FTIR-predicted phenotypes not only for day-to-day herd management, but also as part of a breeding strategy to improve cow performance. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  19. 21 CFR 207.3 - Definitions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... chapter, and any distribution of an animal drug or animal feed bearing or containing an animal drug for..., and animal facilities used for the production or control testing of licensed biologicals, and... definition of drugs in section 201(g) of the act. The term includes manipulation, sampling, testing, or...

  20. 21 CFR 207.3 - Definitions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... chapter, and any distribution of an animal drug or animal feed bearing or containing an animal drug for..., and animal facilities used for the production or control testing of licensed biologicals, and... definition of drugs in section 201(g) of the act. The term includes manipulation, sampling, testing, or...

  1. 21 CFR 207.3 - Definitions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... chapter, and any distribution of an animal drug or animal feed bearing or containing an animal drug for..., and animal facilities used for the production or control testing of licensed biologicals, and... definition of drugs in section 201(g) of the act. The term includes manipulation, sampling, testing, or...

  2. 21 CFR 207.3 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... chapter, and any distribution of an animal drug or animal feed bearing or containing an animal drug for..., and animal facilities used for the production or control testing of licensed biologicals, and... definition of drugs in section 201(g) of the act. The term includes manipulation, sampling, testing, or...

  3. 21 CFR 207.3 - Definitions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... chapter, and any distribution of an animal drug or animal feed bearing or containing an animal drug for..., and animal facilities used for the production or control testing of licensed biologicals, and... definition of drugs in section 201(g) of the act. The term includes manipulation, sampling, testing, or...

  4. Mass spectrometry and animal science: protein identification strategies and particularities of farm animal species.

    PubMed

    Soares, Renata; Franco, Catarina; Pires, Elisabete; Ventosa, Miguel; Palhinhas, Rui; Koci, Kamila; Martinho de Almeida, André; Varela Coelho, Ana

    2012-07-19

    Proteomic approaches are gaining increasing importance in the context of all fields of animal and veterinary sciences, including physiology, productive characterization, and disease/parasite tolerance, among others. Proteomic studies mainly aim the proteome characterization of a certain organ, tissue, cell type or organism, either in a specific condition or comparing protein differential expression within two or more selected situations. Due to the high complexity of samples, usually total protein extracts, proteomics relies heavily on separation procedures, being 2D-electrophoresis and HPLC the most common, as well as on protein identification using mass spectrometry (MS) based methodologies. Despite the increasing importance of MS in the context of animal and veterinary science studies, the usefulness of such tools is still poorly perceived by the animal science community. This is primarily due to the limited knowledge on mass spectrometry by animal scientists. Additionally, confidence and success in protein identification is hindered by the lack of information in public databases for most of farm animal species and their pathogens, with the exception of cattle (Bos taurus), pig (Sus scrofa) and chicken (Gallus gallus). In this article, we will briefly summarize the main methodologies available for protein identification using mass spectrometry providing a case study of specific applications in the field of animal science. We will also address the difficulties inherent to protein identification using MS, with particular reference to experiments using animal species poorly described in public databases. Additionally, we will suggest strategies to increase the rate of successful identifications when working with farm animal species. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Transcriptomic markers meet the real world: finding diagnostic signatures of corticosteroid treatment in commercial beef samples

    PubMed Central

    2012-01-01

    Background The use of growth-promoters in beef cattle, despite the EU ban, remains a frequent practice. The use of transcriptomic markers has already proposed to identify indirect evidence of anabolic hormone treatment. So far, such approach has been tested in experimentally treated animals. Here, for the first time commercial samples were analyzed. Results Quantitative determination of Dexamethasone (DEX) residues in the urine collected at the slaughterhouse was performed by Liquid Chromatography-Mass Spectrometry (LC-MS). DNA-microarray technology was used to obtain transcriptomic profiles of skeletal muscle in commercial samples and negative controls. LC-MS confirmed the presence of low level of DEX residues in the urine of the commercial samples suspect for histological classification. Principal Component Analysis (PCA) on microarray data identified two clusters of samples. One cluster included negative controls and a subset of commercial samples, while a second cluster included part of the specimens collected at the slaughterhouse together with positives for corticosteroid treatment based on thymus histology and LC-MS. Functional analysis of the differentially expressed genes (3961) between the two groups provided further evidence that animals clustering with positive samples might have been treated with corticosteroids. These suspect samples could be reliably classified with a specific classification tool (Prediction Analysis of Microarray) using just two genes. Conclusions Despite broad variation observed in gene expression profiles, the present study showed that DNA-microarrays can be used to find transcriptomic signatures of putative anabolic treatments and that gene expression markers could represent a useful screening tool. PMID:23110699

  6. Development and validation of an immunochromatographic assay for rapid detection of fumonisin B1 from cereal samples.

    PubMed

    Venkataramana, M; Navya, K; Chandranayaka, S; Priyanka, S R; Murali, H S; Batra, H V

    2014-09-01

    Fumonisins are one of the most agriculturally significant environmental toxins produced by Fusarium and Aspergillus species that grow on agricultural commodities in the field or during storage. Cereals contaminated with fumonisins causes serious loss to agricultural produce leads to health problems in humans and other farm animals. In the present study, polyclonal hyperimmune sera was raised against FB1 in rabbits immunized with FB1-keyhole limpet haemocyanin (KLH). Purified antibodies were used to establish a sensitive gold nanoparticle based immunochromatographic strip (ICG) for detecting FB1 levels in cereal grains. Effective on-site detection of FB1 was achieved by developing a rapid and sensitive pAb based ICG strip. This strip had a detection limit of 5 ng mL(-1) for FB1 in cereal samples and it could be completed within 3 min. Close examination of 150 cereal samples by ICG strip method revealed that 77 were fumonisin-positive. Results obtained by the developed method was further validated with well standardized HPLC method and results of strip method was correlated well with those obtained by HPLC method. In conclusion, the developed method was a better alternative for onsite detection of FB1 in cereal samples intended for human consumption to reduce risk of humans and other farm animals. The high level of FB1 concentrations recorded in present study warrants the need to develop an awareness creation programme to the farmers of India for safe handling of cereal grains at the time of harvesting and storage of grains.

  7. Surveillance of Echinococcus isolates from Qinghai, China.

    PubMed

    Ma, Junying; Wang, Hu; Lin, Gonghua; Zhao, Fang; Li, Chao; Zhang, Tongzuo; Ma, Xiao; Zhang, Yongguo; Hou, Zhibin; Cai, Huixia; Liu, Peiyun; Wang, Yongshun

    2015-01-15

    Echinococcosis is highly endemic over large parts of the Qinghai-Tibet Plateau (QTP), China. Based on a large number of samples, we present data on the current presence, host distribution, and genetic diversity of Echinococcus in the Qinghai Province, located in the northeastern corner of the QTP and constituting >25% of the area of the plateau. We used 521 samples (including 451 newly collected samples and 70 samples from our previous study) from humans, yaks, sheep, goats, dogs, fox, plateau pikas, and voles in 36 counties, and genotyped them using the mitochondrial DNA marker cytochrome oxidase subunit I (cox1) gene and the maximum parsimony and Bayesian reconstruction methods. Based on the 792 bp sequence matrix, we recorded 177 variable sites; 157 were parsimony-informative. A total of 105 haplotypes (H1-H105) were detected, of which H1-H15 and H90-H104, H16-H17, H18-H89, and H105 belonged to Echinococcus shiquicus, Echinococcus multilocularis, Echinococcus granulosus, and Echinococcus canadensis, respectively. Our results showed that, (i) the Qinghai Province was under a high burden of Echinococcus epidemiology; (ii) E. granulosus was the main echinococcosis threat to the local people, and the followed is E. multilocularis; (iii) there are a considerable number of haplotypes shared by domestic animals (sheep, yaks, and dogs) and humans, demonstrating the close relationship between human and domestic animals epidemiology; (iv) the threat of E. shiquicus on humans and livestock can be mostly ignored, while the infection risk of E. canadensis echinococcosis should not be neglected. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Development of a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for nitroimidazoles in edible animal tissues and feeds.

    PubMed

    Han, Wei; Pan, Yuanhu; Wang, Yulian; Chen, Dongmei; Liu, Zhenli; Zhou, Qi; Feng, Liang; Peng, Dapeng; Yuan, Zonghui

    2016-02-20

    The misuse of nitroimidazoles (NDZs) can lead to NDZs residues in edible animal tissues, which would be harmful to consumer health. To quickly monitor NDZs residues in edible animal tissues and feed, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) with a simple sample preparation method and clean-up was developed in the present study. At first, a broad-specificity monoclonal antibody, 1D5, against NDZs has been produced, which the IC50 values of the NDZs, dimetridazole, ipronidazole, ronidazole hydroxydimetridazole, and hydroxyipronidazole, were 4.79μgL(-1), 0.47μgL(-1), 5.97μgL(-1), 23.48μgL(-1), and 15.03μgL(-1), respectively. The limit of detection of the method for the NDZ matrix calibration ranged from 4.2μgkg(-1) to 50.3μgkg(-1) in the feed matrices and from 0.11μgkg(-1) to 4.11μgkg(-1) in the edible animal tissues matrices. The recoveries of the NDZs were in the range of 75.5-111.8%. The CVs were less than 14.4%. A good correlation (r=0.9905) between the ELISA and HPLC-MS results of the tissues demonstrated the reliability of the developed ic-ELISA, which makes it a useful tool for screening of NDZs in animal edible tissue and feed. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Water-quality data from ground- and surface-water sites near concentrated animal feeding operations (CAFOs) and non-CAFOs in the Shenandoah Valley and eastern shore of Virginia, January-February, 2004

    USGS Publications Warehouse

    Rice, Karen C.; Monti, Michele M.; Ettinger, Matthew R.

    2005-01-01

    Concentrated animal feeding operations (CAFOs) result from the consolidation of small farms with animals into larger operations, leading to a higher density of animals per unit of land on CAFOs than on small farms. The density of animals and subsequent concentration of animal wastes potentially can cause contamination of nearby ground and surface waters. This report summarizes water-quality data collected from agricultural sites in the Shenandoah Valley and Eastern Shore of Virginia. Five sites, three non-CAFO and two dairy-operation CAFO sites, were sampled in the Shenandoah Valley. Four sites, one non-CAFO and three poultry-operation CAFO sites were sampled on the Eastern Shore. All samples were collected during January and February 2004. Water samples were analyzed for the following parameters and constituents: temperature, specific conductance, pH, and dissolved oxygen; concentrations of the indicator organisms Escherichia coli (E. coli) and enterococci; bacterial isolates of E. coli, enterococci, Salmonella spp., and Campylobacter spp.; sensitivity to antibiotics of E. coli, enterococci, and Salmonella spp.; arsenic, cadmium, chromium3+, copper, nickel, and mercury; hardness, biological oxygen demand, nitrate, nitrite, ammonia, ortho-phosphate, total Kjeldahl nitrogen, chemical oxygen demand, total organic carbon, and dissolved organic carbon; and 45 dissolved organic compounds, which included a suite of antibiotic compounds.Data are presented in tables 5-21 and results of analyses of replicate samples are presented in tables 22-28. A summary of the data in tables 5-8 and 18-21 is included in the report.

  10. [Seroprevalence of tularemia in risk groups of humans and animals in Van, eastern Turkey].

    PubMed

    Bayram, Yasemin; Özkaçmaz, Ayşe; Parlak, Mehmet; Başbuğan, Yıldıray; Kılıç, Selçuk; Güdücüoğlu, Hüseyin

    2015-10-01

    Tularemia has become a re-emerging zoonotic disease in Turkey recently. The aims of this study were to determine the seroprevalence of tularemia in humans and their animals living in rural risky areas of our region and to investigate the risk factors. Between January and July 2012, people living in rural areas of Van province (located at eastern part of Turkey) and their domestic animals were included in the study. The sample size was determined by using cluster sampling method like in an event with known prevalence and planned as a cross-sectional epidemiological study. Proportional random sampling method was used to determine which individuals will be included in the study. Presence of tularemia antibodies in the sera of a total 495 voluntary persons (343 female, 152 male; age range: 18-79 years, mean age: 40.61) and their 171 animals (40 cattle, 124 sheep and 7 goats) were screened by microagglutination test using safranin O-stained F.tularensis antigen (Public Health Agency of Turkey). For the evaluation of cross-reactivity between Brucella spp., tularemia positive serum samples were also tested with brucella microagglutination test. Among human and animal samples, 11.9% (59/495) and 44% (76/171) yielded positive results with the titers of ≥ 1:20 in F.tularensis microagglutination test, respectively. However, 69.5% (41/59) of human sera and 78.9% (60/76) of animal sera demonstrated equal or higher titers in the brucella test, so those sera were considered as cross-reactive. After exclusion of these sera, the seroprevalence for F.tularensis were calculated as 3.6% (18/495) for humans and 9.4% (16/171) for animals. Among the 16 animals with positive results, 12 were sheep, three were cattle and one was goat. The difference between seropositivity rates among the domestic animal species was not statistically significant (p> 0.05). In addition, no statistically significant differences were found between risk factors including insect bite, tick bite, contact with rodents, eating the meat of hunted animals (rabbit), having pet (cat) in home (p> 0.05). In this study, the rate of tularemia seropositivity among humans was similar to the results of previous studies which were performed in our country; however the seropositivity rate of tularemia among domestic animals in our study was higher than the results of a few studies which were conducted on domestic animals. In conclusion, preventive procedures and precautions must be taken into consideration to control the transmission of the infection.

  11. The Effect of Learning Based on Technology Model and Assessment Technique toward Thermodynamic Learning Achievement

    NASA Astrophysics Data System (ADS)

    Makahinda, T.

    2018-02-01

    The purpose of this research is to find out the effect of learning model based on technology and assessment technique toward thermodynamic achievement by controlling students intelligence. This research is an experimental research. The sample is taken through cluster random sampling with the total respondent of 80 students. The result of the research shows that the result of learning of thermodynamics of students who taught the learning model of environmental utilization is higher than the learning result of student thermodynamics taught by simulation animation, after controlling student intelligence. There is influence of student interaction, and the subject between models of technology-based learning with assessment technique to student learning result of Thermodynamics, after controlling student intelligence. Based on the finding in the lecture then should be used a thermodynamic model of the learning environment with the use of project assessment technique.

  12. Adapting Animal-Assisted Therapy Trials to Prison-Based Animal Programs.

    PubMed

    Allison, Molly; Ramaswamy, Megha

    2016-09-01

    Prison-based animal programs have shown promise when it comes to increased sociability, responsibility, and levels of patience for inmates who participate in these programs. Yet there remains a dearth of scientific research that demonstrates the impact of prison-based animal programs on inmates' physical and mental health. Trials of animal-assisted therapy interventions, a form of human-animal interaction therapy most often used with populations affected by depression/anxiety, mental illness, and trauma, may provide models of how prison-based animal program research can have widespread implementation in jail and prison settings, whose populations have high rates of mental health problems. This paper reviews the components of prison-based animal programs most commonly practiced in prisons today, presents five animal-assisted therapy case studies, evaluates them based on their adaptability to prison-based animal programs, and discusses the institutional constraints that act as barriers for rigorous prison-based animal program research implementation. This paper can serve to inform the development of a research approach to animal-assisted therapy that nurses and other public health researchers can use in working with correctional populations. © 2016 Wiley Periodicals, Inc.

  13. [Superficial infections caused by Microsporum canis in humans and animals].

    PubMed

    Segundo, Carolina; Martínez, Alejandrina; Arenas, Roberto; Fernández, Ramón; Cervantes, Roberto A

    2004-03-01

    Dermatophytic infections caused by M. canis in humans and animals have a world wide distribution and they are zoonotic. The objective in this work was to know the frequency of M. canis infections in humans and pets. We studied our cases from January 1994 to December 2002. The human samples were obtained from a Dermatological Department in a General Hospital and we registered the next data: age, sex, job, and affected area. The animal samples were obtained from a mycological veterinary laboratory, and we registered the presence or absence of clinical lesions. A total of 46 clinical cases of M. canis infections were recorded, 26 female and 20 males: tinea capitis 21, tinea corporis 17, tinea pedis five, onychomycosis two, and only one case with tinea faciei. The 46 cases with positive culture yield 42 positive samples in KOH. The age range varied from 2 to 60 years. Among the animals, we studied 461 dogs and found six KOH positive (1%) samples and cultured 23 isolates (4.98%): 21 M. canis, one M. gypseum and one Trichophyton spp. From the 68 samples of cats, eight (11.76%) were positive to KOH, being 26 (38.23%) M. canis isolates. In M. canis infections in humans, the age rage was wide with predominance in women. In animals, M. canis isolates represented the most dermatophytic infection.

  14. Real time lobster posture estimation for behavior research

    NASA Astrophysics Data System (ADS)

    Yan, Sheng; Alfredsen, Jo Arve

    2017-02-01

    In animal behavior research, the main task of observing the behavior of an animal is usually done manually. The measurement of the trajectory of an animal and its real-time posture description is often omitted due to the lack of automatic computer vision tools. Even though there are many publications for pose estimation, few are efficient enough to apply in real-time or can be used without the machine learning algorithm to train a classifier from mass samples. In this paper, we propose a novel strategy for the real-time lobster posture estimation to overcome those difficulties. In our proposed algorithm, we use the Gaussian mixture model (GMM) for lobster segmentation. Then the posture estimation is based on the distance transform and skeleton calculated from the segmentation. We tested the algorithm on a serials lobster videos in different size and lighting conditions. The results show that our proposed algorithm is efficient and robust under various conditions.

  15. A metacognitive illusion in monkeys.

    PubMed

    Ferrigno, Stephen; Kornell, Nate; Cantlon, Jessica F

    2017-09-13

    Like humans, monkeys can make accurate judgements about their own memory by reporting their confidence during cognitive tasks. Some have suggested that animals use associative learning to make accurate confidence judgements, while others have suggested animals directly access and estimate the strength of their memories. Here we test a third, non-exclusive possibility: perhaps monkeys, like humans, base metacognitive inferences on heuristic cues. Humans are known to use cues like perceptual fluency (e.g. how easy something is to see) when making metacognitive judgements. We tested monkeys using a match-to-sample task in which the perceptual fluency of the stimuli was manipulated. The monkeys made confidence wagers on their accuracy before or after each trial. We found that monkeys' wagers were affected by perceptual fluency even when their accuracy was not. This is novel evidence that animals are susceptible to metacognitive illusions similar to those experienced by humans. © 2017 The Author(s).

  16. Defective enamel ultrastructure in diabetic rodents.

    PubMed

    Atar, M; Atar-Zwillenberg, D R; Verry, P; Spornitz, U M

    2004-07-01

    We investigated six different types of diabetic rodents. Four expressed a genetic obesity resulting in diabetes. One developed diabetes induced by a diet-dependent obesity, and one with genetic diabetes received anti-diabetic medication. The tooth samples were examined under a scanning electron microscope and with an energy dispersive microanalysis (EDX). The electron micrographs showed severe, varying degrees of damage within the six different diabetic animal types, such as irregular crystallite deposition and prism perforations in genetically obese animals compared to less-disordered prism structures in diet-dependent obesity. Anti-diabetic medication resulted in normal enamel ultrastructure. The EDX analysis revealed a reduction in the amount of calcium and phosphorus in all regions affected by diabetes. Based on these animal studies, we suggest that both juvenile diabetes type I (in infants) and adult diabetes type II (in pregnant mothers, affecting the developing foetus) may affect the normal development of teeth in humans.

  17. Six Years (2011–2016) of Mandatory Nationwide Bovine Viral Diarrhea Control in Germany—A Success Story

    PubMed Central

    Gethmann, Jörn; Schirrmeier, Horst; Schröder, Ronald; Conraths, Franz J.; Beer, Martin

    2017-01-01

    Bovine viral diarrhea (BVD) is one of the most important infectious diseases in cattle, causing major economic losses worldwide. Therefore, control programs have been implemented in several countries. In Germany, an obligatory nationwide eradication program has been in force since 2011. Its centerpiece is the detection of animals persistently infected (PI) with BVD virus, primarily based on the testing of ear tissue samples of all newborn calves for viral genome or antigen, and their removal from the cattle population. More than 48,000 PI animals have so far been detected and removed. Between the onset of the program and the end of 2016, the prevalence of these animals among all newborn calves decreased considerably, from 0.5% to less than 0.03%. The number of cattle holdings with PI animals likewise decreased from 3.44% in 2011 to only 0.16% in 2016. Since a large number of naïve, fully susceptible animals are now confronted with BVD virus, which is still present in the German cattle population, the challenge of the coming years will be the identification of remaining PI animals as quickly and efficiently as possible, and the efficient protection of BVD-free farms from reinfection. PMID:29057796

  18. Fluid Flow Simulation and Energetic Analysis of Anomalocarididae Locomotion

    NASA Astrophysics Data System (ADS)

    Mikel-Stites, Maxwell; Staples, Anne

    2014-11-01

    While an abundance of animal locomotion simulations have been performed modeling the motions of living arthropods and aquatic animals, little quantitative simulation and reconstruction of gait parameters has been done to model the locomotion of extinct animals, many of which bear little physical resemblance to their modern descendants. To that end, this project seeks to analyze potential swimming patterns used by the anomalocaridid family, (specifically Anomalocaris canadensis, a Cambrian Era aquatic predator), and determine the most probable modes of movement. This will serve to either verify or cast into question the current assumed movement patterns and properties of these animals and create a bridge between similar flexible-bodied swimmers and their robotic counterparts. This will be accomplished by particle-based fluid flow simulations of the flow around the fins of the animal, as well as an energy analysis of a variety of sample gaits. The energy analysis will then be compared to the extant information regarding speed/energy use curves in an attempt to determine which modes of swimming were most energy efficient for a given range of speeds. These results will provide a better understanding of how these long-extinct animals moved, possibly allowing an improved understanding of their behavioral patterns, and may also lead to a novel potential platform for bio-inspired underwater autonomous vehicles (UAVs).

  19. Predicted trends in the supply and demand of veterinarians in Japan.

    PubMed

    Kimura, S; Shinkawa, S; Mago, J; Yamamoto, M; Sakai, M; Sugisaki, T; Karaki, H; Sugiura, K

    2008-12-01

    Currently in Japan, there are 32,000 active veterinarians, mainly engaged in small and large animal practice and public animal health and public health services. In the face of the notable increase in recent years in the proportion of female students enrolled in veterinary schools and in the number of households with companion animals, a model was developed to predict the supply and demand of veterinarians toward 2040 in Japan. Surveys were conducted on sampled households and veterinarians to estimate input variables used in the supply and demand model. From this data it is predicted that there might be somewhere between a shortage of 1,000 to an over-supply of 3,700 veterinarians engaged in small animal practice in 2040. This, however, will depend on possible changes in the number of visits made to veterinarians by small animal owners and the efficiency of practices in the future. The model also predicts that there will be a shortage of around 1,100 veterinarians in large animal practice in 2040. Considering the many assumptions made to estimate the input variables used in the model, the results of this study do not provide definitive conclusions, but provide a base for discussions on what will be needed in the veterinary profession in the future.

  20. The effect of a low essential fatty acid diet on hibernation in marmots.

    PubMed

    Florant, G L; Hester, L; Ameenuddin, S; Rintoul, D A

    1993-04-01

    We investigated the effect of an essential fatty acid (EFA)-deficient diet on hibernation patterns in yellow-bellied marmots (Marmota flaviventris). Fatty acid (FA) analysis of white adipose tissue (WAT) from animals maintained for 2 mo on the EFA-deficient diet suggested that little or no EFAs were present in the gonadal or omental fat depots. Hibernation about lengths of the EFA-deficient animals were significantly shorter (P < 0.01) than control animals. Stated another way, these animals aroused twice as frequently compared with control animals and used more energy to survive winter. Analysis of WAT composition and blood samples revealed that animals were highly lipolytic during winter. Furthermore, the release of FAs was not random: linoleate (cis-9,cis-12-octadecadienoic acid; 18:2, a diene EFA) was significantly (P < 0.05) under-represented in venous outflow from the gonadal WAT pad based on the percentage of this species in WAT. The concentration of saturated FAs was higher than that predicted from the WAT-FA composition. We conclude that linoleate is preferentially retained within WAT and that concentrations of this EFA may influence hibernation behavior. Thus EFAs may have a thermoregulatory role in hibernation in addition to their role as essential precursors for physiologically important lipids after hibernation is over.

  1. Impact assessment and decontamination of pesticides from meat under different culinary processes.

    PubMed

    Sengupta, Dwaipayan; Aktar, Md Wasim; Alam, Samsul; Chowdhury, Ashim

    2010-10-01

    A total of 75 animals between 1.5 and 8 years old were randomly selected for the study. Of these, 57.8% were cross-bred animals and the rest were non-descript. Moreover, 61.8% of the animals under study were brought for slaughter from local sources and the rest from farm houses. Samples collected from five districts revealed contamination with traces of organochlorine pesticides (0.01-0.22 microg g(-1)) and organophosphorus pesticides (0.111-0.098 microg g(-1)). In general, all the raw meat samples possessed dichlorodiphenyltrichloroethane at the highest level. Contamination was highest in cow meat samples and lowest in chicken samples. No particular district-wise trend was obtained for the pesticides selected for analysis. Subsequent decontamination study revealed that cooking is the best option in reducing pesticide load in raw meat samples. Cooked chicken is the safest foodstuff for consumption.

  2. 9 CFR 55.23 - Responsibilities of States and enrolled herd owners.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    .... In cases where animals escape or disappear and thus are not available for tissue sampling and testing, an APHIS representative will investigate whether the unavailability of animals for testing... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Responsibilities of States and...

  3. 9 CFR 55.23 - Responsibilities of States and enrolled herd owners.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    .... In cases where animals escape or disappear and thus are not available for tissue sampling and testing, an APHIS representative will investigate whether the unavailability of animals for testing... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Responsibilities of States and...

  4. 9 CFR 55.23 - Responsibilities of States and enrolled herd owners.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    .... In cases where animals escape or disappear and thus are not available for tissue sampling and testing, an APHIS representative will investigate whether the unavailability of animals for testing... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Responsibilities of States and...

  5. 9 CFR 55.23 - Responsibilities of States and enrolled herd owners.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... testing in accordance with instructions from the APHIS or State representative. In cases where animals... will investigate whether the unavailability of animals or usable samples for testing constitutes a... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Responsibilities of States and...

  6. 9 CFR 55.23 - Responsibilities of States and enrolled herd owners.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... testing in accordance with instructions from the APHIS or State representative. In cases where animals... will investigate whether the unavailability of animals or usable samples for testing constitutes a... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Responsibilities of States and...

  7. Quantifying Individual Variation in the Propensity to Attribute Incentive Salience to Reward Cues

    PubMed Central

    Meyer, Paul J.; Lovic, Vedran; Saunders, Benjamin T.; Yager, Lindsay M.; Flagel, Shelly B.; Morrow, Jonathan D.; Robinson, Terry E.

    2012-01-01

    If reward-associated cues acquire the properties of incentive stimuli they can come to powerfully control behavior, and potentially promote maladaptive behavior. Pavlovian incentive stimuli are defined as stimuli that have three fundamental properties: they are attractive, they are themselves desired, and they can spur instrumental actions. We have found, however, that there is considerable individual variation in the extent to which animals attribute Pavlovian incentive motivational properties (“incentive salience”) to reward cues. The purpose of this paper was to develop criteria for identifying and classifying individuals based on their propensity to attribute incentive salience to reward cues. To do this, we conducted a meta-analysis of a large sample of rats (N = 1,878) subjected to a classic Pavlovian conditioning procedure. We then used the propensity of animals to approach a cue predictive of reward (one index of the extent to which the cue was attributed with incentive salience), to characterize two behavioral phenotypes in this population: animals that approached the cue (“sign-trackers”) vs. others that approached the location of reward delivery (“goal-trackers”). This variation in Pavlovian approach behavior predicted other behavioral indices of the propensity to attribute incentive salience to reward cues. Thus, the procedures reported here should be useful for making comparisons across studies and for assessing individual variation in incentive salience attribution in small samples of the population, or even for classifying single animals. PMID:22761718

  8. Research on animation design of growing plant based on 3D MAX technology

    NASA Astrophysics Data System (ADS)

    Chen, Yineng; Fang, Kui; Bu, Weiqiong; Zhang, Xiaoling; Lei, Menglong

    In view of virtual plant has practical demands on quality, image and degree of realism animation in growing process of plant, this thesis design the animation based on mechanism and regularity of plant growth, and propose the design method based on 3D MAX technology. After repeated analysis and testing, it is concluded that there are modeling, rendering, animation fabrication and other key technologies in the animation design process. Based on this, designers can subdivid the animation into seed germination animation, plant growth prophase animation, catagen animation, later animation and blossom animation. This paper compounds the animation of these five stages by VP window to realize the completed 3D animation. Experimental result shows that the animation can realized rapid, visual and realistic simulatation the plant growth process.

  9. [Establishment of an Animal Based Therapy at a University Hospital for Psychiatry: Results of a Preliminary Study and Future Prospects].

    PubMed

    Hartfiel, Cajetan; Bodatsch, Mitja; Klosterkötter, Joachim; Kuhn, Jens

    2017-01-01

    Objective: Dogs have been integrated in human society over centuries. This process has selected unique social and communicative skills. Dogs are thus able to represent social substitutes for human counterparts in cases of social withdrawal. Furthermore, dogs act as "social catalysts" in promoting interhuman encounters. Thus, the integration of dogs in psychotherapeutic concepts addressing social and interpersonal deficits may be of special interest. Methods: The present investigation reports the results of a pilot study to establish animal-assisted therapy (dogs) at a psychiatric department. The animal-assisted intervention straddled the following areas: (1) contact making, communication and orientation to needs, (2) recreation and play, (3) outward orientation, (4) release and farewell. The sample comprised 22 subjects. Results: The results demonstrated in particular that the animal-assisted intervention significantly promoted unspecific aspects of positive affectivity and wellbeing. Evaluation of the overall acceptance of the dog on the psychiatric ward revealed very positive feedback. Conclusions: We conclude that animal-assisted therapies represent a significant enchrichment of the therapeutic context, that may be used to enhance the patients' openness and adherence to conventional therapies. © Georg Thieme Verlag KG Stuttgart · New York.

  10. Forage nutritive value of plant samples collected by clipping or rumen evacuation

    USDA-ARS?s Scientific Manuscript database

    Knowing nutritive value and intake of forage by an animal is important to understand methane production data from individual animals in a grazed environment. Digestibility of forages is typically evaluated as the in vitro dry matter disappearance (IVDMD) using a clipped forage sample; however, beef...

  11. Assessing the robustness of quantitative fatty acid signature analysis to assumption violations

    USGS Publications Warehouse

    Bromaghin, Jeffrey F.; Budge, Suzanne M.; Thiemann, Gregory W.; Rode, Karyn D.

    2016-01-01

    In most QFASA applications, investigators will generally have some knowledge of the prey available to predators and be able to assess the completeness of prey signature data and sample additional prey as necessary. Conversely, because calibration coefficients are derived from feeding trials with captive animals and their values may be sensitive to consumer physiology and nutritional status, their applicability to free-ranging animals is difficult to establish. We therefore recommend that investigators first make any improvements to the prey signature data that seem warranted and then base estimation on the Aitchison distance measure, as it appears to minimize risk from violations of the assumption that is most difficult to verify.

  12. Analysis of survival data from telemetry projects

    USGS Publications Warehouse

    Bunck, C.M.; Winterstein, S.R.; Pollock, K.H.

    1985-01-01

    Telemetry techniques can be used to study the survival rates of animal populations and are particularly suitable for species or settings for which band recovery models are not. Statistical methods for estimating survival rates and parameters of survival distributions from observations of radio-tagged animals will be described. These methods have been applied to medical and engineering studies and to the study of nest success. Estimates and tests based on discrete models, originally introduced by Mayfield, and on continuous models, both parametric and nonparametric, will be described. Generalizations, including staggered entry of subjects into the study and identification of mortality factors will be considered. Additional discussion topics will include sample size considerations, relocation frequency for subjects, and use of covariates.

  13. Non-invasive assessment of adrenocortical function in captive Nile crocodiles (Crocodylus niloticus).

    PubMed

    Ganswindt, Stefanie B; Myburgh, Jan G; Cameron, Elissa Z; Ganswindt, Andre

    2014-11-01

    The occurrence of stress-inducing factors in captive crocodilians is a concern, since chronic stress can negatively affect animal health and reproduction, and hence production. Monitoring stress in wild crocodiles could also be beneficial for assessing the state of health in populations which are potentially threatened by environmental pollution. In both cases, a non-invasive approach to assess adrenocortical function as a measure of stress would be preferable, as animals are not disturbed during sample collection, and therefore sampling is feedback-free. So far, however, such a non-invasive method has not been established for any crocodilian species. As an initial step, we therefore examined the suitability of two enzyme-immunoassays, detecting faecal glucocorticoid metabolites (FGMs) with a 11β,21-diol-20-one and 5β-3α-ol-11-one structure, respectively, for monitoring stress-related physiological responses in captive Nile crocodiles (Crocodylus niloticus). An adrenocorticotropic hormone (ACTH) challenge was performed on 10 sub-adult crocodiles, resulting in an overall increase in serum corticosterone levels of 272% above the pre-injection levels 5h post-injection. Saline-treated control animals (n=8) showed an overall increase of 156% in serum corticosterone levels 5h post-administration. Faecal samples pre- and post-injection could be obtained from three of the six individually housed crocodiles, resulting in FGM concentrations 136-380% above pre-injection levels, always detected in the first sample collected post-treatment (7-15 days post-injection). FGM concentrations seem comparatively stable at ambient temperatures for up to 72 h post-defaecation. In conclusion, non-invasive hormone monitoring can be used for assessing adrenocortical function in captive Nile crocodiles based on FGM analysis. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Ultraviolet-C irradiation for inactivation of viruses in foetal bovine serum.

    PubMed

    Vaidya, Vivek; Dhere, Rajeev; Agnihotri, Snehal; Muley, Ravindra; Patil, Sanjay; Pawar, Amit

    2018-07-05

    Foetal Bovine Serum (FBS) and porcine trypsin are one of the essential raw materials used in the manufacturing of cell culture based viral vaccines. Being from animal origin, these raw materials can potentially contaminate the final product by known or unknown adventitious agents. The issue is more serious in case of live attenuated viral vaccines, where there is no inactivation step which can take care of such adventitious agents. It is essential to design production processes which can offer maximum viral clearance potential for animal origin products. Ultraviolet-C irradiation is known to inactivate various adventitious viral agents; however there are limited studies on ultraviolet inactivation of viruses in liquid media. We obtained a recently developed UVivatec ultraviolet-C (UV-C) irradiation based viral clearance system for evaluating its efficacy to inactivate selected model viruses. This system has a unique design with spiral path of liquid allowing maximum exposure to UV-C light of a short wavelength of 254 nm. Five live attenuated vaccine viruses and four other model viruses were spiked in tissue culture media and exposed to UV-C irradiation. The pre and post UV-C irradiation samples were analyzed for virus content to find out the extent of inactivation of various viruses. These experiments showed substantial log reduction for the majority of the viruses with few exceptions based on the characteristics of these viruses. Having known the effect of UV irradiation on protein structure, we also evaluated the post irradiation samples of culture media for growth promoting properties using one of the most fastidious human diploid cells (MRC-5). UV-C exposure did not show any notable impact on the nutritional properties of culture media. The use of an UV-C irradiation based system is considered to be promising approach to mitigate the risk of adventitious agents in cell culture media arising through animal derived products. Copyright © 2018 Elsevier Ltd. All rights reserved.

  15. Prevalence of Mycobacterium avium subspecies paratuberculosis and hepatitis E in New World camelids in Austria.

    PubMed

    Stanitznig, A; Khol, J L; Lambacher, B; Franz, S; Wittek, T; Kralik, P; Slana, I; Vasickova, P

    2017-07-07

    Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of paratuberculosis in domestic ruminants and New World Camelids (NWC). Hepatitis E virus (HEV) is an important public health concern worldwide. The virus has been identified in several species, some of them serving as a reservoir for zoonotic HEV strains. Husbandry and breeding of llamas and alpacas have increased in Austria in recent years. Therefore, the aim of the present study was to evaluate the prevalence of MAP and HEV in NWC in Austria. Altogether 445 animals, originating from 78 farms were enrolled in the study. Of the animals sampled, 184 (41.35%) were llamas and 261 (58.65%) were alpacas. 443 blood samples for MAP-ELISA and 399 faecal samples for quantitative PCR (qPCR) and culture for MAP as well as for HEV detection by RT-qPCR have been collected. All of the 399 animals tested for shedding of MAP were negative by faecal solid culture. Using qPCR, 15 (3.8%) of the animals were MAP positive and 384 (96.2%) negative. Out of the 443 serum samples examined for specific antibodies against MAP by ELISA, 6 (1.4%) were positive, 1 (0.2%) was questionable and 436 (98.4%) samples were negative. All faecal samples were tested negative for HEV.

  16. Occurrence of male-specific bacteriophage in feral and domestic animal wastes, human feces, and human-associated wastewaters.

    PubMed

    Calci, K R; Burkhardt, W; Watkins, W D; Rippey, S R

    1998-12-01

    Male-specific bacteriophage (MSB) densities were determined in animal and human fecal wastes to assess their potential impact on aquatic environments. Fecal samples (1,031) from cattle, chickens, dairy cows, dogs, ducks, geese, goats, hogs, horses, seagulls, sheep, and humans as well as 64 sewerage samples were examined for MSB. All animal species were found to harbor MSB, although the great majority excreted these viruses at very low levels. The results from this study demonstrate that in areas affected by both human and animal wastes, wastewater treatment plants are the principal contributors of MSB to fresh, estuarine, and marine waters.

  17. Occurrence of Male-Specific Bacteriophage in Feral and Domestic Animal Wastes, Human Feces, and Human-Associated Wastewaters

    PubMed Central

    Calci, Kevin R.; Burkhardt, William; Watkins, William D.; Rippey, Scott R.

    1998-01-01

    Male-specific bacteriophage (MSB) densities were determined in animal and human fecal wastes to assess their potential impact on aquatic environments. Fecal samples (1,031) from cattle, chickens, dairy cows, dogs, ducks, geese, goats, hogs, horses, seagulls, sheep, and humans as well as 64 sewerage samples were examined for MSB. All animal species were found to harbor MSB, although the great majority excreted these viruses at very low levels. The results from this study demonstrate that in areas affected by both human and animal wastes, wastewater treatment plants are the principal contributors of MSB to fresh, estuarine, and marine waters. PMID:9835602

  18. OBT analysis method using polyethylene beads for limited quantities of animal tissue.

    PubMed

    Kim, S B; Stuart, M

    2015-08-01

    This study presents a polyethylene beads method for OBT determination in animal tissues and animal products for cases where the amount of water recovered by combustion is limited by sample size or quantity. In the method, the amount of water recovered after combustion is enhanced by adding tritium-free polyethylene beads to the sample prior to combustion in an oxygen bomb. The method reduces process time by allowing the combustion water to be easily collected with a pipette. Sufficient water recovery was achieved using the polyethylene beads method when 2 g of dry animal tissue or animal product were combusted with 2 g of polyethylene beads. Correction factors, which account for the dilution due to the combustion water of the beads, are provided for beef, chicken, pork, fish and clams, as well as egg, milk and cheese. The method was tested by comparing its OBT results with those of the conventional method using animal samples collected on the Chalk River Laboratories (CRL) site. The results determined that the polyethylene beads method added no more than 25% uncertainty when appropriate correction factors are used. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

  19. Detection of rotavirus species A, B and C in domestic mammalian animals with diarrhoea and genotyping of bovine species A rotavirus strains.

    PubMed

    Otto, Peter H; Rosenhain, Stefanie; Elschner, Mandy C; Hotzel, Helmut; Machnowska, Patrycja; Trojnar, Eva; Hoffmann, Kathrin; Johne, Reimar

    2015-09-30

    Rotaviruses (RVs) are a major cause of neonatal diarrhoea in humans and animals worldwide. In this study, 425 faecal samples were collected between 1999 and 2013 from diarrhoeic livestock and companion animals at different locations in Germany and tested for RVs. A previously published real-time RT-PCR assay was optimized for detection of a larger variety of RV species A (RVA) strains, and real-time RT-PCR assays for detection of RV species B (RVB) and C (RVC) were newly developed. The detection limits of the assays were 1.54×10(2), 3.95×10(2) and 3.60×10(3) genome copies for RVA, RVB and RVC, respectively. RVA was identified in 85.2% of bovine samples, 51.2% of porcine samples, 50.0% of feline samples, 43.2% of equine samples and 39.7% of canine samples. RVB was found in 3.0% of bovine samples, 2.7% of equine samples and 1.6% of porcine samples. RVC was detected in 31.0% of porcine samples, 21.7% of feline samples, 9.0% of canine samples and 6.0% of bovine samples. For genotyping, 101 RVA-positive bovine samples were further analysed by semi-nested RT-PCR. Genotype combination G6P[5] was most frequently detected (67.3% of samples), followed by G6P[11] (13.9%), G10P[5] (4.0%), G8P[11] (3.0%), G6P[1] (1.0%), and G10P[11] (1.0%). Mixed RVA infections were detected in 5.9% of samples; no or incomplete typing was possible in 4.0% of the samples. This first overview on RV species and RVA genotypes in diarrhoeic livestock and companion animals from Germany indicates a broad circulation of a large variety of RVs. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Detection of Ebola Virus RNA through Aerosol Sampling of Animal Biosafety Level 4 Rooms Housing Challenged Nonhuman Primates

    DTIC Science & Technology

    2016-08-02

    301- 619-4768(f). 1 2 3 4 5 6 7 8 Title: Detection of Ebola Virus RNA through Aerosol Sampling of Animal Biosafety Level 9 4...5 6 To whom it may concern, 7 8 My colleagues and I are submitting the attached manuscript, Detection of Ebola Virus 9 RNA through Aerosol...embedded in the texts. This is the first report demonstrating detection of Ebola virus 17 RNA from animal rooms housing infected nonhuman primates and

  1. Survey of Francisella tularensis in Wild Animals in Japan in Areas Where Tularemia is Endemic.

    PubMed

    Hotta, Akitoyo; Tanabayashi, Kiyoshi; Fujita, Osamu; Shindo, Junji; Park, Chu-Ho; Kudo, Noboru; Hatai, Hitoshi; Oyamada, Toshifumi; Yamamoto, Yoshie; Takano, Ai; Kawabata, Hiroki; Sharma, Neekun; Uda, Akihiko; Yamada, Akio; Morikawa, Shigeru

    2016-09-21

    Samples taken from 428 wild animals and 126 ticks, collected from a tularemia-endemic area in Japan between 2005 and 2013, were analyzed for the presence of Francisella tularensis. F. tularensis was isolated from a Japanese hare carcass whereas the samples from live animals and ticks were negative for F. tularensis by real-time PCR. Our results suggest that F. tularensis is still present in Japan although its prevalence is considerably low even in areas where tularemia is endemic.

  2. Epidemiological patterns of bovine besnoitiosis in an endemic beef cattle herd reared under extensive conditions.

    PubMed

    Esteban-Gil, A; Calvete, C; Casasús, I; Sanz, A; Ferrer, J; Peris, M P; Marcén-Seral, J M; Castillo, J A

    2017-03-15

    Bovine besnoitiosis is a parasitic disease caused by the protozoan Besnoitia besnoiti. Described many decades ago, recent epidemiological studies reveal its important spread within Europe in the last years. To date, many epidemiological aspects related to life cycle, routes of transmission, incidence rates and associated risk factors are lacking; hence, the establishment of appropriate disease control programmes poses an important challenge. Thus, the aim of the present study was to determine the epidemiological pattern of the disease in an endemic herd reared under extensive conditions (Spanish Pyrenees) by identifying main factors associated with infection and clinical disease dynamics. The study population consisted of 276 Brown Swiss and Pirenaica adult animals and 145 calves born and weaned at the farm during the study. Three sampling time frames were used: January 2010, September 2010 and February 2011, which allowed us to differentiate two periods designated as mountain and valley periods. The data related to animals (breed, sex and age) and herd management (animal grouping and time in housing) were recorded. The data collection methodology was mainly based on clinical examinations and defining the serological status against bovine besnoitiosis by the immunofluorescent antibody testing of blood samples. The total prevalence among adult animals was 38.34% (CI95%: 34.53-42.07), with 18.54% of seropositive animals showing clinical signs. In regard to the cumulative incidence, 34.57% of new infections were detected during the mountain period, in contrast to the 24.59% observed in the valley period. The incidence density was 0.058 and 0.061 new infections per animal-month for the mountain and valley periods, respectively. According to the seroepidemiological study, the seroconversion probability of B. besnoiti infection was directly associated with the number of seropositive cows with whom an animal had been stabled as well as the housing period duration, supporting horizontal transmission by close contact as one of the most important methods of disease spread. In addition, the risk of developing the clinical course increased with age, and the presence of clinical signs was related to higher antibody responses. Among calves (from 3.1 to 7.1 months old) sampled once at weaning, the total seroprevalence was 15.17% (CI95%: 9.36-21.04), and the chronic stage was observed in three animals, supporting the ability of B. besnoiti to infect and even cause disease in animals less than 6 months old. Finally, the risk of calf seroconversion was positively related to the serological status of the cows, suggesting postnatal transmission between dams and offspring by contact during the suckling period. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Characterizing biological variability in livestock blood cholinesterase activity for biomonitoring organophosphate nerve agent exposure

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

    Halbrook, R.S.; Shugart, L.R.; Watson, A.P.

    1992-09-01

    A biomonitoring protocol, using blood cholinesterase (ChE) activity in livestock as a monitor of potential organophosphate nerve agent exposure during the planned destruction of US unitary chemical warfare agent stockpiles, is described. The experimental design included analysis of blood ChE activity in individual healthy sheep, horses, and dairy and beef cattle during a 10- to 12-month period. Castrated and sexually intact males, pregnant and lactating females, and adult and immature animals were examined through at least one reproductive cycle. The same animals were used throughout the period of observation and were not exposed to ChE-inhibiting organophosphate or carbamate compounds. Amore » framework for an effective biomonitoring protocol within a monitoring area includes establishing individual baseline blood ChE activity for a sentinel group of 6 animals on the bases of blood samples collected over a 6-month period, monthly collection of blood samples for ChE-activity determination during monitoring, and selection of adult animals as sentinels. Exposure to ChE-inhibiting compounds would be suspected when all blood ChE activity of all animals within the sentinel group are decreased greater than 20% from their own baseline value. Sentinel species selection is primarily a logistical and operational concern; however, sheep appear to be the species of choice because within-individual baseline ChE activity and among age and gender group ChE activity in sheep had the least variability, compared with data from other species. This protocol provides an effective and efficient means for detecting abnormal depressions in blood ChE activity in livestock and can serve as a valuable indicator of the extent of actual plume movement and/or deposition in the event of organophosphate nerve agent release.« less

  4. Detection of porcine DNA in gelatine and gelatine-containing processed food products-Halal/Kosher authentication.

    PubMed

    Demirhan, Yasemin; Ulca, Pelin; Senyuva, Hamide Z

    2012-03-01

    A commercially available real-time PCR, based on a multi-copy target cytochrome b (cyt b) using porcine specific primers, has been validated for the Halal/Kosher authentication of gelatine. Extraction and purification of DNA from gelatine were successfully achieved using the SureFood® PREP Animal system, and real-time PCR was carried out using SureFood® Animal ID Pork Sens kit. The minimum level of adulteration that could be detected was 1.0% w/w for marshmallows and gum drops. A small survey was undertaken of processed food products such as gum drops, marshmallows and Turkish delight, believed to contain gelatine. Of fourteen food products from Germany, two samples were found to contain porcine gelatine, whereas of twenty-nine samples from Turkey twenty-eight were negative. However, one product from Turkey contained porcine DNA and thus was not Halal, and neither was the use of porcine gelatine indicated on the product label. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Magnetic graphene dispersive solid phase extraction combining high performance liquid chromatography for determination of fluoroquinolones in foods.

    PubMed

    He, Xin; Wang, Geng Nan; Yang, Kun; Liu, Hui Zhi; Wu, Xia Jun; Wang, Jian Ping

    2017-04-15

    In this study, a magnetic graphene-based dispersive solid phase extraction method was developed that was combined with high performance liquid chromatography to determine the residues of fluoroquinolone drugs in foods of animal origin. During the experiments, several parameters possible influencing the extraction performance were optimized (amount of magnetic graphene, sample pH, extraction time and elution solution). This extraction method showed high absorption capacities (>6800ng) and high enrichment factors (68-79-fold) for seven fluoroquinolones. Furthermore, this absorbent could be reused for at least 40 times. The limits of detection were in the range of 0.05-0.3ng/g, and the recoveries from the standards fortified blank samples (bovine milk, chicken muscle and egg) were in the range of 82.4-108.5%. Therefore, this method could be used as a simple and sensitive tool to determine the residues of fluoroquinolones in foods of animal origin. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Application of DNA forensic techniques for identifying poached guanacos (Lama guanicoe) in Chilean Patagonia*.

    PubMed

    Marín, Juan C; Saucedo, Cristian E; Corti, Paulo; González, Benito A

    2009-09-01

    Guanaco (Lama guanicoe) is a protected and widely distributed ungulate in South America. A poacher, after killing guanacos in Valle Chacabuco, Chilean Patagonia, transported and stored the meat. Samples were retrieved by local police but the suspect argued that the meat was from a horse. Mitochondrial cytochrome b gene (774 pb), 15 loci microsatellites, and SRY gene were used to identify the species, number of animals and their population origin, and the sex of the animals, respectively. Analysis revealed that the samples came from a female (absence of SRY gene) Patagonian guanaco (assignment probability between 0.0075 and 0.0282), and clearly distinguishing it from sympatric ungulates (E-value = 0). Based on the evidence obtained in the field in addition to forensic data, the suspect was convicted of poaching and illegally carrying fire arms. This is the first report of molecular tools being used in forensic investigations of Chilean wildlife indicating its promising future application in guanaco management and conservation.

  7. The prevalence of Coxiella burnetii infection in wild Korean water deer, Korea.

    PubMed

    Shin, Gee-Wook; Kim, Eun-Ju; Lee, Hae-Beom; Cho, Ho-Seong

    2014-07-01

    The aim of this study was to evaluate the prevalence of Coxiella burnetti infection in wild Korean water deer (Hydropotes inermis argyropus) in Korea, by using serology and real-time PCR analyses. One hundred ninety-six sera were collected from 4 provinces and tested for anti-C. burnetii antibody detection, by means of CHEKIT Q fever ELISA kit; and C. burnetii IS1111 insertion sequence detection, by means of real-time PCR. Antibodies were detected in 18 of the 196 (9.18%) serum samples, whereas genomes of C. burnetii were detected in 13 of the 196 (6.63%) serum samples. Based on overall high seroprevalence, the public health implications of these findings are important, because they indicate that asymptomatic seropositive or seronegative wild animals may be consistently shedding C. burnetii. This is the first study of C. burnetii prevalence in Korean water deer in the Republic of Korea that has indicated the presence of infected animals throughout the country.

  8. The Prevalence of Coxiella burnetii Infection in Wild Korean Water Deer, Korea

    PubMed Central

    SHIN, Gee-Wook; KIM, Eun-Ju; LEE, Hae-Beom; CHO, Ho-Seong

    2014-01-01

    ABSTRACT The aim of this study was to evaluate the prevalence of Coxiella burnetti infection in wild Korean water deer (Hydropotes inermis argyropus) in Korea, by using serology and real-time PCR analyses. One hundred ninety-six sera were collected from 4 provinces and tested for anti-C. burnetii antibody detection, by means of CHEKIT Q fever ELISA kit; and C. burnetii IS1111 insertion sequence detection, by means of real-time PCR. Antibodies were detected in 18 of the 196 (9.18%) serum samples, whereas genomes of C. burnetii were detected in 13 of the 196 (6.63%) serum samples. Based on overall high seroprevalence, the public health implications of these findings are important, because they indicate that asymptomatic seropositive or seronegative wild animals may be consistently shedding C. burnetii. This is the first study of C. burnetii prevalence in Korean water deer in the Republic of Korea that has indicated the presence of infected animals throughout the country. PMID:24717533

  9. Contamination with bacterial zoonotic pathogen genes in U.S. streams influenced by varying types of animal agriculture.

    PubMed

    Haack, Sheridan K; Duris, Joseph W; Kolpin, Dana W; Focazio, Michael J; Meyer, Michael T; Johnson, Heather E; Oster, Ryan J; Foreman, William T

    2016-09-01

    Animal waste, stream water, and streambed sediment from 19 small (<32km(2)) watersheds in 12U.S. states having either no major animal agriculture (control, n=4), or predominantly beef (n=4), dairy (n=3), swine (n=5), or poultry (n=3) were tested for: 1) cholesterol, coprostanol, estrone, and fecal indicator bacteria (FIB) concentrations, and 2) shiga-toxin producing and enterotoxigenic Escherichia coli, Salmonella, Campylobacter, and pathogenic and vancomycin-resistant enterococci by polymerase chain reaction (PCR) on enrichments, and/or direct quantitative PCR. Pathogen genes were most frequently detected in dairy wastes, followed by beef, swine and poultry wastes in that order; there was only one detection of an animal-source-specific pathogen gene (stx1) in any water or sediment sample in any control watershed. Post-rainfall pathogen gene numbers in stream water were significantly correlated with FIB, cholesterol and coprostanol concentrations, and were most highly correlated in dairy watershed samples collected from 3 different states. Although collected across multiple states and ecoregions, animal-waste gene profiles were distinctive via discriminant analysis. Stream water gene profiles could also be discriminated by the watershed animal type. Although pathogen genes were not abundant in stream water or streambed samples, PCR on enrichments indicated that many genes were from viable organisms, including several (shiga-toxin producing or enterotoxigenic E. coli, Salmonella, vancomycin-resistant enterococci) that could potentially affect either human or animal health. Pathogen gene numbers and types in stream water samples were influenced most by animal type, by local factors such as whether animals had stream access, and by the amount of local rainfall, and not by studied watershed soil or physical characteristics. Our results indicated that stream water in small agricultural U.S. watersheds was susceptible to pathogen gene inputs under typical agricultural practices and environmental conditions. Pathogen gene profiles may offer the potential to address both source of, and risks associated with, fecal pollution. Published by Elsevier B.V.

  10. Developmental vitamin D deficiency and risk of schizophrenia: a 10-year update.

    PubMed

    McGrath, John J; Burne, Thomas H; Féron, François; Mackay-Sim, Allan; Eyles, Darryl W

    2010-11-01

    There is an urgent need to generate and test candidate risk factors that may explain gradients in the incidence of schizophrenia. Based on clues from epidemiology, we proposed that developmental vitamin D deficiency may contribute to the risk of developing schizophrenia. This hypothesis may explain diverse epidemiological findings including season of birth, the latitude gradients in incidence and prevalence, the increased risk in dark-skinned migrants to certain countries, and the urban-rural gradient. Animal experiments demonstrate that transient prenatal hypovitaminosis D is associated with persisting changes in brain structure and function, including convergent evidence of altered dopaminergic function. A recent case-control study based on neonatal blood samples identified a significant association between neonatal vitamin D status and risk of schizophrenia. This article provides a concise summary of the epidemiological and animal experimental research that has explored this hypothesis.

  11. Occurrence of human pathogenic Clostridium botulinum among healthy dairy animals: an emerging public health hazard.

    PubMed

    Abdel-Moein, Khaled A; Hamza, Dalia A

    2016-01-01

    The current study was conducted to investigate the occurrence of human pathogenic Clostridium botulinum in the feces of dairy animals. Fecal samples were collected from 203 apparently healthy dairy animals (50 cattle, 50 buffaloes, 52 sheep, 51 goats). Samples were cultured to recover C. botulinum while human pathogenic C. botulinum strains were identified after screening of all C. botulinum isolates for the presence of genes that encode toxins type A, B, E, F. The overall prevalence of C. botulinum was 18.7% whereas human pathogenic C. botulinum strains (only type A) were isolated from six animals at the rates of 2, 2, 5.8, and 2% for cattle, buffaloes, sheep, and goats, respectively. High fecal carriage rates of C. botulinum among apparently healthy dairy animals especially type A alarm both veterinary and public health communities for a potential role which may be played by dairy animals in the epidemiology of such pathogen.

  12. Risk-based approach to developing a national residue sampling plan for testing under European Union regulation for veterinary medicinal products and coccidiostat feed additives in domestic animal production.

    PubMed

    Danaher, Martin; Shanahan, Conor; Butler, Francis; Evans, Rhodri; O'Sullivan, Dan; Glynn, Denise; Camon, Tim; Lawlor, Peadar; O'Keeffe, Michael

    2016-07-01

    A ranking system for veterinary medicinal products and coccidiostat feed additives has been developed as a tool to be applied in a risk-based approach to the residue testing programme for foods of animal origin in the Irish National Residue Control Plan (NRCP). Three characteristics of substances that may occur as residues in food are included in the developed risk ranking system: Potency, as measured by the acceptable daily intake assigned by the European Medicines Agency Committee for Medicinal Products for Veterinary Use, to each substance; Usage, as measured by the three factors of Number of Doses, use on Individual animals or for Group treatment, and Withdrawal Period; and Residue Occurrence, as measured by the number of Non-Compliant Samples in the NRCP. For both Number of Doses and Non-Compliant Samples, data for the 5-year period 2008-12 have been used. The risk ranking system for substances was developed for beef cattle, sheep and goats, pigs, chickens and dairy cattle using a scoring system applied to the various parameters described above to give an overall score based on the following equation: Potency × Usage (Number of Doses + Individual/Group Use + Withdrawal Period) × Residue Occurrence. Applying this risk ranking system, the following substances are ranked very highly: antimicrobials such as amoxicillin (for all species except pigs), marbofloxacillin (for beef cattle), oxytetracycline (for all species except chickens), sulfadiazine with trimethoprim (for pigs and chickens) and tilmicosin (for chickens); antiparasitic drugs, such as the benzimidazoles triclabendazole (for beef and dairy cattle), fenbendazole/oxfendazole (for sheep/goats and dairy cattle) and albendazole (for dairy cattle), the avermectin ivermectin (for beef cattle), and anti-fluke drugs closantel and rafoxanide (for sheep/goats); the anticoccidials monensin, narasin, nicarbazin and toltrazuril (for chickens). The risk ranking system described is a relatively simple system designed to provide a reliable basis for selecting the veterinary medicinal products and coccidiostat feed additives that might be prioritised for residue testing.

  13. Iophenoxic acid derivatives as markers of oral baits to wildlife. New tools for their detection in tissues of a game species and safety considerations for human exposure.

    PubMed

    Sage, Mickael; Fourel, Isabelle; Lahoreau, Jennifer; Siat, Vivien; Berny, Philippe; Rossi, Sophie

    2013-05-01

    The bait-marker iophenoxic acid (IPA) and its derivatives are increasingly used for evaluating and optimizing the cost-effectiveness of baiting campaigns on wildlife, particularly on game species such as the wild boar. We aimed to determine whether concentrations of the three main IPA derivatives ethyl, methyl and propyl-IPA measured on thoracic liquid extracts (TLE) of hunted wild boars may be representative of two exposure doses, 40 and 200 mg, from 20 to 217 days after ingestion. Then we developed a method of detection of the three IPA derivatives by LC/ESI-MS-MS in muscle and liver to evaluate the suitability of these two other tissues for monitoring the marked bait consumption and for measuring available residues in the meat of marked animals. Three semi-captive wild boars received 40 mg of each IPA derivative, three received 200 mg, and three, as controls, did not receive IPA. Blood serum was sampled 20, 197 or 217 days after IPA exposure according to animals and to the derivative. Wild boars were shot by gun after the different times of serum sampling times, and TLE, muscle and liver were sampled. Our results suggest that TLE is not a relevant tissue for quantitatively expressing IPA exposure. Due to interference, no analytical method was validated on TLE containing digestive material. On the other hand, quantifications in the muscle and particularly in the liver could discriminate wild boars that had ingested the two IPA doses from 20 days until 7 months after exposure, especially for the two long term markers ethyl and propyl-IPA. So IPA quantifications in the liver sampled on hunted animals appear to be a reliable tool for monitoring bait consumption in the field at a large scale. Nevertheless, whatever the ingested dose, ethyl- and propyl-IPA concentrations measured in the muscle and the liver of tested animals until 217 days after exposure, remained higher than 0.01 mg/kg, the Maximal Residue Limit (MRL) is recommended for molecules for which no toxicological data are available. Based on the range of IPA residues available in these two tissues, implications for humans consuming marked animals are discussed.

  14. Culture-based indicators of fecal contamination and molecular microbial indicators rarely correlate with Campylobacter spp. in recreational waters.

    PubMed

    Hellein, Kristen N; Battie, Cynthia; Tauchman, Eric; Lund, Deanna; Oyarzabal, Omar A; Lepo, Joe Eugene

    2011-12-01

    Campylobacter spp. are the leading cause of gastroenteritis worldwide. Most human infections result from contaminated food; however, infections are also caused by recreational waterway contamination. Campylobacter culture is technically challenging and enumeration by culture-based methods is onerous. Thus, we employed qPCR to quantify Campylobacter spp. in fresh- and marine-water samples, raw sewage and animal feces. Multiplex PCR determined whether Campylobacter jejuni or C. coli, most commonly associated with human disease, were present in qPCR-positive samples. Campylobacters were detected in raw sewage, and in feces of all avian and mammalian species tested. Campylobacter-positive concentrations ranged from 68 to 2.3 × 10⁶ cells per 500 mL. Although C. jejuni and C. coli were rare in waterways, they were prevalent in sewage and feces. Campylobacter-specific qPCR screening of environmental waters did not correlate with the regulatory EPA method 1600 (Enterococcus culture), nor with culture-independent, molecular-based microbial source tracking indicators, such as human polyomavirus, human Bacteroidales and Methanobrevibacter smithii. Our results suggest that neither the standard EPA method nor the newly proposed culture-independent methods are appropriate surrogates for Campylobacter contamination in water. Thus, assays for specific pathogens may be necessary to protect human health, especially in waters that are contaminated with sewage and animal feces.

  15. Horvitz-Thompson survey sample methods for estimating large-scale animal abundance

    USGS Publications Warehouse

    Samuel, M.D.; Garton, E.O.

    1994-01-01

    Large-scale surveys to estimate animal abundance can be useful for monitoring population status and trends, for measuring responses to management or environmental alterations, and for testing ecological hypotheses about abundance. However, large-scale surveys may be expensive and logistically complex. To ensure resources are not wasted on unattainable targets, the goals and uses of each survey should be specified carefully and alternative methods for addressing these objectives always should be considered. During survey design, the impoflance of each survey error component (spatial design, propofiion of detected animals, precision in detection) should be considered carefully to produce a complete statistically based survey. Failure to address these three survey components may produce population estimates that are inaccurate (biased low), have unrealistic precision (too precise) and do not satisfactorily meet the survey objectives. Optimum survey design requires trade-offs in these sources of error relative to the costs of sampling plots and detecting animals on plots, considerations that are specific to the spatial logistics and survey methods. The Horvitz-Thompson estimators provide a comprehensive framework for considering all three survey components during the design and analysis of large-scale wildlife surveys. Problems of spatial and temporal (especially survey to survey) heterogeneity in detection probabilities have received little consideration, but failure to account for heterogeneity produces biased population estimates. The goal of producing unbiased population estimates is in conflict with the increased variation from heterogeneous detection in the population estimate. One solution to this conflict is to use an MSE-based approach to achieve a balance between bias reduction and increased variation. Further research is needed to develop methods that address spatial heterogeneity in detection, evaluate the effects of temporal heterogeneity on survey objectives and optimize decisions related to survey bias and variance. Finally, managers and researchers involved in the survey design process must realize that obtaining the best survey results requires an interactive and recursive process of survey design, execution, analysis and redesign. Survey refinements will be possible as further knowledge is gained on the actual abundance and distribution of the population and on the most efficient techniques for detection animals.

  16. Sampling animal sign in heterogeneous environments: how much is enough?

    USGS Publications Warehouse

    Holbrook, Joseph D.; Arkle, Robert S.; Rachlow, Janet L.; Vierling, Kerri T.; Pilliod, David S.

    2015-01-01

    Animal ecologists often use animal sign as a surrogate for direct observation of organisms, especially when species are secretive or difficult to observe. Spatial heterogeneity in arid environments makes it challenging to consistently detect and precisely characterize animal sign, which can bias estimates of animal abundance or habitat use. Piute ground squirrels (Urocitellus mollis) and Owyhee harvester ants (Pogonomyrmex salinus) live in arid environments and are fossorial, which can make them difficult to observe directly. Their relative abundance can be assessed using sign (i.e., burrows and nests). We implemented an over-sampling framework (i.e., recorded an excessive amount of information) with two observers to 1) identify a sampling intensity that balanced precision with our resource constraints, and 2) assess classification and detection of squirrel burrows and ant nests across vegetation conditions. We sampled 20 1-ha plots for ground squirrel burrows and ant nests using six 4 m × 100 m belt transects. Analyses of precision and sampling effort indicated that three belt transects covering 1200 m2 per ha provided sufficient precision, while minimizing effort. Regardless of vegetation conditions, counts by two observers were strongly correlated for ground squirrel burrows (r = 0.99, P < 0.001, df = 18; slope = 0.92) and harvester ant nests (r = 0.99, P < 0.001, df = 18; slope = 1.01) indicating observer consistency and perhaps high detection probability. These findings illustrate an approach for evaluating sampling designs in many ecological contexts.

  17. Comparative costs of the Mouse Inoculation Test (MIT) and Virus Isolation in Cell Culture (VICC) for use in rabies diagnosis in Brazil.

    PubMed

    Bones, Vanessa C; Gameiro, Augusto H; Castilho, Juliana G; Molento, Carla F M

    2015-05-01

    The decision to use laboratory animals rather than in vitro methods is frequently based on the financial costs involved, so the objective of our study was to compare the costs of performing the Mouse Inoculation Test (MIT) and Virus Isolation in Cell Culture (VICC) for use in rabies diagnosis in Brazil. Based on observations of laboratory routines at the Pasteur Institute, São Paulo, we listed the fixed cost (FC) and variable cost (VC) items necessary to perform both tests. Considering that 200 MITs are equivalent to 350 VICC assays, in terms of facilities and staff-hours needed per month, we calculated, for both tests, the average total cost per sample, the costs of the implementation of the laboratory structure, and the costs of routine use. With regard to absolute values, the total cost was mainly influenced by FC items, as they represented 60% of the cost for the MIT and 86% of the cost for VICC. A sample analysed by the MIT costs around 205% more than one analysed by using VICC. The MIT costs 74% and 406% more than VICC, when implementation costs and routine use per month, respectively, are taken into account. Our results can assist in the resolution of costing disputes that could hinder the replacement of animals for rabies diagnosis in Brazil. The method demonstrated here might also be useful for cost comparisons in other situations where animal use still continues when validated alternatives exist. 2015 FRAME.

  18. In vivo biodistribution of CNTs using a BALB/c mouse experimental model.

    PubMed

    Fufă, Mariana Oana Mihaela; Mihaiescu, Dan Eduard; Mogoantă, Laurenţiu; Bălşeanu, Tudor Adrian; Mogoşanu, George Dan; Grumezescu, Alexandru Mihai; Bolocan, Alexandra

    2015-01-01

    Due to their unique behaviors, carbon nanotubes (CNTs)-based systems meet essential requirements for modern applications, such as electronics, optics, photovoltaics, fuel cells, aerospace engineering, military and biomedical applications. CNTs biocompatibility and toxic effects were assessed both in vitro and in vivo, in terms of hemocompatibility, cytocompatibility, immunoreactions and genetic behavior. The aim of this paper is to evaluate the in vivo biodistribution and biocompatibility of carbon nanopowder synthesized by plasma processing, using a BALB/c mouse experimental model. Three months old BALB/c mice were aseptically injected with 100 μL of 1 mg/mL dispersions. The obtained carbon-based nano-systems were dispersed in saline solution and subsequently sterilized by using a 30 minutes treatment with UV irradiation. The reference mice were injected with 100 μL of saline. The mice were kept under standard conditions of light, temperature, humidity, food and water (ad libitum) before the vital organ harvest. The animal welfare was daily monitored. At two and 10 days after the inoculation, the animals were euthanized under general anesthesia, for the sampling of internal organs (brain, myocardium, pancreas, liver, lung, kidney and spleen). No animal died during the experiment. Brain, myocardium and pancreas were histologically normal, with no tissue damage, inflammatory infiltrate or inorganic deposits. CNTs were evidenced only in hepatic, renal, pulmonary and spleen tissue samples. Increased amounts of inorganic granular structures were reported after 10 days of treatment, when compared to the short-term (two days) inoculation. Our BALB/c mouse experimental model was found to be useful for the in vivo assessment of biodistribution and biocompatibility of CNTs.

  19. Method for sampling and analysis of volatile biomarkers in process gas from aerobic digestion of poultry carcasses using time-weighted average SPME and GC-MS.

    PubMed

    Koziel, Jacek A; Nguyen, Lam T; Glanville, Thomas D; Ahn, Heekwon; Frana, Timothy S; Hans van Leeuwen, J

    2017-10-01

    A passive sampling method, using retracted solid-phase microextraction (SPME) - gas chromatography-mass spectrometry and time-weighted averaging, was developed and validated for tracking marker volatile organic compounds (VOCs) emitted during aerobic digestion of biohazardous animal tissue. The retracted SPME configuration protects the fragile fiber from buffeting by the process gas stream, and it requires less equipment and is potentially more biosecure than conventional active sampling methods. VOC concentrations predicted via a model based on Fick's first law of diffusion were within 6.6-12.3% of experimentally controlled values after accounting for VOC adsorption to the SPME fiber housing. Method detection limits for five marker VOCs ranged from 0.70 to 8.44ppbv and were statistically equivalent (p>0.05) to those for active sorbent-tube-based sampling. The sampling time of 30min and fiber retraction of 5mm were found to be optimal for the tissue digestion process. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Point prevalence of Cryptosporidium oocyst in calves grazing along River Rima bank in Sokoto, Nigeria.

    PubMed

    Faleke, O O; Yabo, Y A; Olaleye, A O; Dabai, Y U; Ibitoye, E B

    2014-02-01

    The present study was conducted to investigate the point prevalence of Cryptosporidium oocysts infection in calves grazing along the bank of Rima River Sokoto in October 2011. The river bank is a converging zone for domestic animals reared in different quarters of the town and the surrounding settlements. A total number of 2,959 cattle were enumerated out of which 147 (4.97%) were calves. Faecal samples were collected from 100 (68.02%) calves by convenient sampling technique. Formol-Ether sedimentation and modified Ziehl-Neelsen staining techniques were used to identify the Cryptosporidium oocysts in the faecal samples. Faecal consistency was also used to identify diarrhoeic and non-diarrhoeic calves. Cryptosporidium oocysts were identified in 33 (33.0%) of the calves examined. The detection rate was higher among the male calves (38.46%) than females while the Rahaji breed had the highest prevalence of 62.5%. A total of 6 (18.18%) among the positive cases were diarrhoeic. The differences in prevalence based on sex, breeds and presence of diarrhoea were not statistically significant. Calves may become sources of Cryptosporidia infection to man and other animals in the study area through unrestricted movements and interactions with the environment.

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