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Sample records for paepalanthus spp extracts

  1. The Andean Paepalanthus pilosus complex (Eriocaulaceae): a revision with three new taxa

    PubMed Central

    Hensold, Nancy

    2016-01-01

    Abstract A herbarium-based revision is provided for Paepalanthus pilosus and allies, five commonly confused species of cushion plants native to Andean paramo. These are placed in the recircumscribed Paepalanthus subsect. Cryptanthella Suess. The group includes Paepalanthus pilosus, Paepalanthus dendroides, and Paepalanthus lodiculoides. An additional two species and one variety are newly described: Paepalanthus caryonauta, Paepalanthus huancabambensis, and Paepalanthus pilosus var. leoniae. The latter two are Peruvian endemics, while Paepalanthus caryonauta is known from four countries, and has long been confused with other species. An additional, possibly undescribed taxon is noted from the Serrania de Perijá, Colombia. Five new synonyms and three lectotypes are proposed, and the common misapplication of some names is noted. Within the Paepalanthus pilosus complex, species differences were found in timing of peduncle elongation, sex ratio, and leaf, perianth, diaspore and nectary morphology. Ecological differences are suggested by specimen data and a review of ecological literature. Descriptions, photographs and maps are provided for all species, as is a key to the groups of eriocaulaceous cushion plants from Andean South America. PMID:27489483

  2. The Andean Paepalanthus pilosus complex (Eriocaulaceae): a revision with three new taxa.

    PubMed

    Hensold, Nancy

    2016-01-01

    A herbarium-based revision is provided for Paepalanthus pilosus and allies, five commonly confused species of cushion plants native to Andean paramo. These are placed in the recircumscribed Paepalanthus subsect. Cryptanthella Suess. The group includes Paepalanthus pilosus, Paepalanthus dendroides, and Paepalanthus lodiculoides. An additional two species and one variety are newly described: Paepalanthus caryonauta, Paepalanthus huancabambensis, and Paepalanthus pilosus var. leoniae. The latter two are Peruvian endemics, while Paepalanthus caryonauta is known from four countries, and has long been confused with other species. An additional, possibly undescribed taxon is noted from the Serrania de Perijá, Colombia. Five new synonyms and three lectotypes are proposed, and the common misapplication of some names is noted. Within the Paepalanthus pilosus complex, species differences were found in timing of peduncle elongation, sex ratio, and leaf, perianth, diaspore and nectary morphology. Ecological differences are suggested by specimen data and a review of ecological literature. Descriptions, photographs and maps are provided for all species, as is a key to the groups of eriocaulaceous cushion plants from Andean South America. PMID:27489483

  3. A SIMPLE METHOD FOR THE EXTRACTION AND QUANTIFICATION OF PHOTOPIGMENTS FROM SYMBIODINIUM SPP.

    EPA Science Inventory

    John E. Rogers and Dragoslav Marcovich. Submitted. Simple Method for the Extraction and Quantification of Photopigments from Symbiodinium spp.. Limnol. Oceanogr. Methods. 19 p. (ERL,GB 1192).

    We have developed a simple, mild extraction procedure using methanol which, when...

  4. In vitro anticancer activity of extracts of Mentha Spp. against human cancer cells.

    PubMed

    Sharma, Vikas; Hussain, Shabir; Gupta, Moni; Saxena, Ajit Kumar

    2014-10-01

    In vitro anticancer potential of methanolic and aqueous extracts of whole plants of Mentha arvensis, M. longifolia, M. spicata and M. viridis at concentration of 100 μg/ml was evaluated against eight human cancer cell lines--A-549, COLO-205, HCT-116, MCF-7, NCI-H322, PC-3, THP-1 and U-87MG from six different origins (breast, colon, glioblastoma, lung, leukemia and prostate) using sulphorhodamine blue (SRB) assay. Methanolic extracts of above-mentioned Mentha Spp. displayed anti-proliferative effect in the range of 70-97% against four human cancer cell lines, namely COLO-205, MCF-7, NCI-H322 and THP-1; however, aqueous extracts were found to be active against HCT-116 and PC-3. The results indicate that Mentha Spp. contain certain constituents with cytotoxic properties which may find use in developing anticancer agents.

  5. DNA extraction methods and multiple sampling to improve molecular diagnosis of Sarcocystis spp. in cattle hearts.

    PubMed

    Bräunig, Patrícia; Portella, Luiza Pires; Cezar, Alfredo Skrebsky; Libardoni, Felipe; Sangioni, Luis Antonio; Vogel, Fernanda Silveira Flores; Gonçalves, Paulo Bayard Dias

    2016-10-01

    Molecular detection of Sarcocystis spp. in tissue samples can be useful for experimental and diagnostic purposes. However, the parasite spreads unevenly through tissues, forming tissue cysts, and the cystic wall is an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the cysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of four protocols for DNA extraction from cysts of Sarcocystis spp. present in bovine myocardium samples or after their harvest in phosphate-buffered saline (PBS) solution as well as determine the effects of single or multiple sampling on the accuracy of molecular diagnosis of sarcocystosis in cattle hearts. Cysts and myocardium samples from nine bovine hearts were randomly distributed to four DNA extraction protocols: kit, kit with modification, DNAzol, and cetyl-trimethyl ammonium bromide (CTAB). Samples were submitted to DNA extraction and PCR as replicates of each heart (simplicate, duplicate, and triplicate), and the probability of a true positive diagnostic was calculated. Among the protocols tested, the kit with modification was determined to be the most suitable for DNA extraction from cysts in PBS solution (92.6 % of DNA detection by PCR); DNAzol resulted in higher DNA detection frequency from bovine myocardium samples (48.1 %). Multiple sampling improved the molecular diagnosis of Sarcocystis spp. infection in cattle hearts, increasing at 22.2 % the rate of true positive diagnostic.

  6. Allelopathic effect of Bromus spp. and Lolium spp. shoot extracts on some crops.

    PubMed

    Lehoczky, E; Nelima, M Okumu; Szabó, R; Szalai, A; Nagy, P

    2011-01-01

    Allelopathy is an untapped resource for weed control in crops that could give good possibilities for environmentally sound, integrated crop production. Allelopathy is defined as the direct or indirect harmful or beneficial effects of one plant on another through the production of chemical compounds, called allelochemicals, which escape into the environment. Allelochemicals can be produced by weeds and affect crops, and the reverse is also true. Allelopathic interactions include weed-weed, weed-crop, and crop-crop. Allelopathy offers potential for selective biological weed control for instance weed-suppressing crops and the use of plant residues in cropping systems, allelopathic rotational crops, or companion plants with allelopathic potential. Bromus species occur in many habitats in temperate regions of the world, including America, Eurasia, Australia, and Africa. The genus Lolium is one of the most important forage grasses. The weed species usually grow in the same production zones as wheat and are considered weeds since they parasitize wheat fields. Some of the weed species in these two genus have been reported to have allelopathic effect. One of the methods that has been successful in studying allelopathic activity are bioassays. Laboratory experiments were conducted to determine allelopathic effect of watery shoot extracts of four weed species of the Poaceae family, namely Bromus rigidus, Bromus diandrus, Lolium multiflorum and Lolium temulentum on germination and growth of winter wheat (Triticum aestivum L.), spring barley (Hordeum vulgare L.), corn (Zea mays L), perennial ryegrass (Lolium perenne L.), bean (Phaseolus sp.) and sunflower (Helianthus annuus L.) and on each other. The experiment was carried out during the period March 2010 to October 2010. Twenty five seeds were put into one Petri-dish on filter paper, adding 15ml of extract to each in four repeats. The germination took place in a Binder-type thermostat in the dark. The timing of germination was

  7. Allelopathic effect of Bromus spp. and Lolium spp. shoot extracts on some crops.

    PubMed

    Lehoczky, E; Nelima, M Okumu; Szabó, R; Szalai, A; Nagy, P

    2011-01-01

    Allelopathy is an untapped resource for weed control in crops that could give good possibilities for environmentally sound, integrated crop production. Allelopathy is defined as the direct or indirect harmful or beneficial effects of one plant on another through the production of chemical compounds, called allelochemicals, which escape into the environment. Allelochemicals can be produced by weeds and affect crops, and the reverse is also true. Allelopathic interactions include weed-weed, weed-crop, and crop-crop. Allelopathy offers potential for selective biological weed control for instance weed-suppressing crops and the use of plant residues in cropping systems, allelopathic rotational crops, or companion plants with allelopathic potential. Bromus species occur in many habitats in temperate regions of the world, including America, Eurasia, Australia, and Africa. The genus Lolium is one of the most important forage grasses. The weed species usually grow in the same production zones as wheat and are considered weeds since they parasitize wheat fields. Some of the weed species in these two genus have been reported to have allelopathic effect. One of the methods that has been successful in studying allelopathic activity are bioassays. Laboratory experiments were conducted to determine allelopathic effect of watery shoot extracts of four weed species of the Poaceae family, namely Bromus rigidus, Bromus diandrus, Lolium multiflorum and Lolium temulentum on germination and growth of winter wheat (Triticum aestivum L.), spring barley (Hordeum vulgare L.), corn (Zea mays L), perennial ryegrass (Lolium perenne L.), bean (Phaseolus sp.) and sunflower (Helianthus annuus L.) and on each other. The experiment was carried out during the period March 2010 to October 2010. Twenty five seeds were put into one Petri-dish on filter paper, adding 15ml of extract to each in four repeats. The germination took place in a Binder-type thermostat in the dark. The timing of germination was

  8. Comparison of three phenotypic and deoxyribonucleic acid extraction methods for isolation and Identification of Nocardia spp

    PubMed Central

    Faghri, Jamshid; Bourbour, Samane; Moghim, Sharare; Meidani, Mohsen; Safaei, Hajiye Ghasemian; Hosseini, Nafise; Esfahani, Bahram Nasr; Fazeli, Hussein; Sedighi, Mansour

    2014-01-01

    Background: The aerobic Actinomycetes are a large group of soil-indwelling bacteria that are distributed in world-wide. These Gram-positive bacteria are most commonly associated with opportunistic infections in both immunocompromised and immunocompetent hosts. Materials and Methods: In this study, three phenotypic and deoxyribonucleic acid (DNA) extraction methods for isolation and identification of Nocardia genus were compared. Samples were taken in five different locations of Isfahan's suburb from hospitals area, parks, agricultural lands, gardens, arid lands with different soil temperature and pH. Results: In this study, showed that slip-buried-method was better than two other phenotypic methods; 14 out of 70 soil samples (20%) were positive for Nocardia spp. DNA of positive samples were extracted with three techniques and DNA extraction by microwave technique was better than others. This technique was confirmed with observation of DNA bands on 1% agarose gel. Conclusions: These bacteria are important in immune deficient patients such as cancer patients, transplant recipients, tuberculosis; acquired immunodeficiency syndrome etc., Their affluence is unsteady in different zones of the world. In this study, among the three phenotypic methods for the isolation of Nocardia slip-buried method was better than other methods. Among DNA extraction techniques, DNA extraction by microwave method would be selective method for DNA extraction of Nocardia spp. compared with others techniques. PMID:25221754

  9. DNA extraction-free quantification of Dehalococcoides spp. in groundwater using a hand-held device.

    PubMed

    Stedtfeld, Robert D; Stedtfeld, Tiffany M; Kronlein, Maggie; Seyrig, Gregoire; Steffan, Robert J; Cupples, Alison M; Hashsham, Syed A

    2014-12-01

    Nucleic acid amplification of biomarkers is increasingly used to measure microbial activity and predict remedial performance in sites with trichloroethene (TCE) contamination. Field-based genetic quantification of microorganisms associated with bioremediation may help increase accuracy that is diminished through transport and processing of groundwater samples. Sterivex cartridges and a previously undescribed mechanism for eluting biomass was used to concentrate cells. DNA extraction-free loop mediated isothermal amplification (LAMP) was monitored in real-time with a point of use device (termed Gene-Z). A detection limit of 10(5) cells L(–1) was obtained, corresponding to sensitivity between 10 to 100 genomic copies per reaction for assays targeting the Dehalococcoides spp. specific 16S rRNA gene and vcrA gene, respectively. The quantity of Dehalococcoides spp. genomic copies measured from two TCE contaminated groundwater samples with conventional means of quantification including filtration, DNA extraction, purification, and qPCR was comparable to the field ready technique. Overall, this method of measuring Dehalococcoides spp. and vcrA genes in groundwater via direct amplification without intentional DNA extraction and purification is demonstrated, which may provide a more accurate mechanism of predicting remediation rates.

  10. Antimicrobial activity of wax and hexane extracts from Citrus spp. peels.

    PubMed

    Johann, Susana; Oliveira, Vetúria Lopes de; Pizzolatti, Moacir G; Schripsema, Jan; Braz-Filho, Raimundo; Branco, Alexsandro; Smânia Jr, Artur

    2007-09-01

    Antibacterial and antifungal properties of wax and hexane extracts of Citrus spp. peels were tested using bioautographic and microdilution techniques against three plant pathogenic fungi (Penicillium digitatum, Curvularia sp., and Colletotrichum sp.), two human pathogens (Trichophyton mentagrophytes and Microsporum canis), and two opportunistic bacteria (Escherichia coli and Staphylococcus aureus). Two polymethoxylated flavonoids and a coumarin derivative, were isolated and identified from peel extracts, which presented antimicrobial activity especially against M. canis and T. mentagrophytes: 4',5,6,7,8-pentamethoxyflavone (tangeritin) and 3',4',5,6,7,8-hexamethoxyflavone (nobiletin) from C. reticulata; and 6,7-dimethoxycoumarin (also known as escoparone, scoparone or scoparin) from C. limon. PMID:17923995

  11. Supercritical Carbon Dioxide Extraction of Carotenoids from Pumpkin (Cucurbita spp.): A Review

    PubMed Central

    Durante, Miriana; Lenucci, Marcello Salvatore; Mita, Giovanni

    2014-01-01

    Carotenoids are well known for their nutritional properties and health promoting effects representing attractive ingredients to develop innovative functional foods, nutraceutical and pharmaceutical preparations. Pumpkin (Cucurbita spp.) flesh has an intense yellow/orange color owing to the high level of carotenoids, mainly α-carotene, β-carotene, β-cryptoxanthin, lutein and zeaxanthin. There is considerable interest in extracting carotenoids and other bioactives from pumpkin flesh. Extraction procedures able to preserve nutritional and pharmacological properties of carotenoids are essential. Conventional extraction methods, such as organic solvent extraction (CSE), have been used to extract carotenoids from plant material for a long time. In recent years, supercritical carbon dioxide (SC-CO2) extraction has received a great deal of attention because it is a green technology suitable for the extraction of lipophylic molecules and is able to give extracts of high quality and totally free from potentially toxic chemical solvents. Here, we review the results obtained so far on SC-CO2 extraction efficiency and quali-quantitative composition of carotenoids from pumpkin flesh. In particular, we consider the effects of (1) dehydration pre-treatments; (2) extraction parameters (temperature and pressure); the use of water, ethanol and olive oil singularly or in combination as entrainers or pumpkin seeds as co-matrix. PMID:24756094

  12. Supercritical carbon dioxide extraction of carotenoids from pumpkin (Cucurbita spp.): a review.

    PubMed

    Durante, Miriana; Lenucci, Marcello Salvatore; Mita, Giovanni

    2014-04-21

    Carotenoids are well known for their nutritional properties and health promoting effects representing attractive ingredients to develop innovative functional foods, nutraceutical and pharmaceutical preparations. Pumpkin (Cucurbita spp.) flesh has an intense yellow/orange color owing to the high level of carotenoids, mainly α-carotene, β-carotene, β-cryptoxanthin, lutein and zeaxanthin. There is considerable interest in extracting carotenoids and other bioactives from pumpkin flesh. Extraction procedures able to preserve nutritional and pharmacological properties of carotenoids are essential. Conventional extraction methods, such as organic solvent extraction (CSE), have been used to extract carotenoids from plant material for a long time. In recent years, supercritical carbon dioxide (SC-CO2) extraction has received a great deal of attention because it is a green technology suitable for the extraction of lipophylic molecules and is able to give extracts of high quality and totally free from potentially toxic chemical solvents. Here, we review the results obtained so far on SC-CO2 extraction efficiency and quali-quantitative composition of carotenoids from pumpkin flesh. In particular, we consider the effects of (1) dehydration pre-treatments; (2) extraction parameters (temperature and pressure); the use of water, ethanol and olive oil singularly or in combination as entrainers or pumpkin seeds as co-matrix.

  13. Amebicidal activity of plant extracts from Southeast Asia on Acanthamoeba spp.

    PubMed

    Chu, D M; Miles, H; Toney, D; Ngyuen, C; Marciano-Cabral, F

    1998-09-01

    The effect of 100 polar and 100 nonpolar plant extract materials obtained from Southeast Asia were evaluated for amebicidal activity in vitro against three species of Acanthamoeba. A. culbertsoni, A. castellanii, and A. polyphaga, the causative agents of granulomatous amebic encephalitis and amebic keratitis, were studied in vitro to determine whether the plant extracts exhibited amebicidal activity or induced encystment of the amebae. Of the 200 plant extracts tested, extracts obtained from three plants (Ipomoea sp., Kaempferia galanga, and Cananga odorata) were amebicidal for all three species of Acanthamoeba and a fourth extract prepared from Gastrochilus panduratum was lytic for A. polyphaga and growth-inhibitory for A. castellanii and A. culbertsoni. Three plant extracts induced encystment of all three species of Acanthamoeba. Select plant extracts were tested as well for tumoricidal activity against B103 neuroblastoma cells. Some plant extracts that exhibited tumoricidal activity for B103 cells were not amebicidal for Acanthamoeba spp. Additionally, the polar and nonpolar extracts that exhibited amebicidal activity were also tested for activity against primary murine peritoneal macrophage cultures. Plant extracts that demonstrated tumoricidal or amebicidal activity were not lytic for normal macrophage cultures. PMID:9766904

  14. Comparison of Different Protein Extraction Methods for Gel-Based Proteomic Analysis of Ganoderma spp.

    PubMed

    Al-Obaidi, Jameel R; Saidi, Noor Baity; Usuldin, Siti Rokhiyah Ahmad; Hussin, Siti Nahdatul Isnaini Said; Yusoff, Noornabeela Md; Idris, Abu Seman

    2016-04-01

    Ganoderma species are a group of fungi that have the ability to degrade lignin polymers and cause severe diseases such as stem and root rot and can infect economically important plants and perennial crops such as oil palm, especially in tropical countries such as Malaysia. Unfortunately, very little is known about the complex interplay between oil palm and Ganoderma in the pathogenesis of the diseases. Proteomic technologies are simple yet powerful tools in comparing protein profile and have been widely used to study plant-fungus interaction. A critical step to perform a good proteome research is to establish a method that gives the best quality and a wide coverage of total proteins. Despite the availability of various protein extraction protocols from pathogenic fungi in the literature, no single extraction method was found suitable for all types of pathogenic fungi. To develop an optimized protein extraction protocol for 2-DE gel analysis of Ganoderma spp., three previously reported protein extraction protocols were compared: trichloroacetic acid, sucrose and phenol/ammonium acetate in methanol. The third method was found to give the most reproducible gels and highest protein concentration. Using the later method, a total of 10 protein spots (5 from each species) were successfully identified. Hence, the results from this study propose phenol/ammonium acetate in methanol as the most effective protein extraction method for 2-DE proteomic studies of Ganoderma spp. PMID:27016942

  15. Antinociceptive Activity and Effect of Methanol Extracts of Three Salvia Spp. on Withdrawal Syndrome in Mice

    PubMed Central

    Karami, Mohammad; Shamerani, Mohammad Mehdi; Hossini, Ebrahim; Gohari, Ahmad Reza; Ebrahimzadeh, Mohammad Ali; Nosrati, Anahita

    2013-01-01

    Purpose: There are several reports about effects of Salvia spp. on CNS. The present experiment is undertaken to study effect of S. limbata, S. hypoleuca and S. macrosiphon on withdrawal syndrome in mice. Methods: Antinociceptive activities of aerial parts of Salvia spp. is investigated using hot plate method. In addition, the effect of its aerial parts on morphine dependence is investigated in mice. After induction of morphine dependency, different concentrations of plant extract are injected. To assess morphine withdrawal, naloxone (5 mg kg-1, i.p.) are injected into mice on the 5th day. Withdrawal syndrome is assessed by placing each mouse in a glass box 30 cm in height and recording the incidence of escape jumps for 60 minutes. Results: A decrease in incidence of escape jumps is observed in morphine dependence mice. S. limbata and S. hypoleuca extracts produced a statistically significant inhibition of pain induced by hot plate latency at (500, 1000 and 1500 mg kg-1) i.p. A significant increase in pain threshold is observed after 30 and 60 minutes (p < 0.001). The activity was comparable to that of morphine (30 mg kg-1, i.p., p > 0.05). The antinociceptive activity increased up to 60 minutes. Conclusion: S. limbataand S. hypolecuca extracts produced statistically significant inhibition of pain and development of morphine dependence in mice. PMID:24312878

  16. Efficacy of antimicrobials extracted from organic pecan shell for inhibiting the growth of Listeria spp.

    PubMed

    Babu, Dinesh; Crandall, Philip G; Johnson, Casey L; O'Bryan, Corliss A; Ricke, Steven C

    2013-12-01

    Growers and processors of USDA certified organic foods are in need of suitable organic antimicrobials. The purpose of the research reported here was to develop and test natural antimicrobials derived from an all-natural by-product, organic pecan shells. Unroasted and roasted organic pecan shells were subjected to solvent free extraction to produce antimicrobials that were tested against Listeria spp. and L. monocytogenes serotypes to determine the minimum inhibitory concentrations (MIC) of antimicrobials. The effectiveness of pecan shell extracts were further tested using a poultry skin model system and the growth inhibition of the Listeria cells adhered onto the skin model were quantified. The solvent free extracts of pecan shells inhibited Listeria strains at MICs as low as 0.38%. The antimicrobial effectiveness tests on a poultry skin model exhibited nearly a 2 log reduction of the inoculated cocktail mix of Listeria strains when extracts of pecan shell powder were used. The extracts also produced greater than a 4 log reduction of the indigenous spoilage bacteria on the chicken skin. Thus, the pecan shell extracts may prove to be very effective alternative antimicrobials against food pathogens and supplement the demand for effective natural antimicrobials for use in organic meat processing.

  17. Efficacy of antimicrobials extracted from organic pecan shell for inhibiting the growth of Listeria spp.

    PubMed

    Babu, Dinesh; Crandall, Philip G; Johnson, Casey L; O'Bryan, Corliss A; Ricke, Steven C

    2013-12-01

    Growers and processors of USDA certified organic foods are in need of suitable organic antimicrobials. The purpose of the research reported here was to develop and test natural antimicrobials derived from an all-natural by-product, organic pecan shells. Unroasted and roasted organic pecan shells were subjected to solvent free extraction to produce antimicrobials that were tested against Listeria spp. and L. monocytogenes serotypes to determine the minimum inhibitory concentrations (MIC) of antimicrobials. The effectiveness of pecan shell extracts were further tested using a poultry skin model system and the growth inhibition of the Listeria cells adhered onto the skin model were quantified. The solvent free extracts of pecan shells inhibited Listeria strains at MICs as low as 0.38%. The antimicrobial effectiveness tests on a poultry skin model exhibited nearly a 2 log reduction of the inoculated cocktail mix of Listeria strains when extracts of pecan shell powder were used. The extracts also produced greater than a 4 log reduction of the indigenous spoilage bacteria on the chicken skin. Thus, the pecan shell extracts may prove to be very effective alternative antimicrobials against food pathogens and supplement the demand for effective natural antimicrobials for use in organic meat processing. PMID:24279287

  18. Ultrasound-assisted green solvent extraction of high-added value compounds from microalgae Nannochloropsis spp.

    PubMed

    Parniakov, O; Apicella, E; Koubaa, M; Barba, F J; Grimi, N; Lebovka, N; Pataro, G; Ferrari, G; Vorobiev, E

    2015-12-01

    The aim of this work was to investigate ultrasound (US)-assisted green solvent extraction of valuable compounds from the microalgae Nannochloropsis spp. Individual green solvents (water, ethanol (EtOH), dimethyl sulfoxide (DMSO)) and binary mixture of solvents (water-DMSO and water-EtOH) were used for the extraction procedures. Maximum total phenolic compounds yield (Yp ≈ 0.33) was obtained after US pre-treatment (W=400 W, 15 min), being almost 5-folds higher compared to that found for the untreated samples and aqueous extraction (Yp ≈ 0.06). The highest yield of total chlorophylls (Yc ≈ 0.043) was obtained after US (W=400 W, 7.5 min), being more than 9-folds higher than those obtained for the untreated samples and aqueous extraction (Yc ≈ 0.004). The recovery efficiency decreased as DMSO>EtOH>H2O. The optimal conditions to recover phenolic compounds and chlorophylls from microalgae were obtained after US pre-treatment (400 W, 5 min), binary mixtures of solvents (water-DMSO and water-EtOH) at 25-30%, and microalgae concentration of 10%. PMID:26398670

  19. High-antibacterial activity of Urtica spp. seed extracts on food and plant pathogenic bacteria.

    PubMed

    Körpe, Didem Aksoy; İşerı, Özlem Darcansoy; Sahin, Feride Iffet; Cabi, Evren; Haberal, Mehmet

    2013-05-01

    The aim of this study was to comparatively evaluate antibacterial activities of methanol (MetOH) and aqueous (dw) leaf (L), root (R) and seed (S) extracts of Urtica dioica L. (Ud; stinging nettle) and Urtica pilulifera L. (Up; Roman nettle) on both food- and plant-borne pathogens, with total phenolic contents and DPPH radical scavenging activities (DRSA). MetOH extracts of leaves and roots of U. dioica had the highest DRSA. Extracts with high antibacterial activity were in the order Up-LMetOH (13/16) > Ud-SMetOH (11/16) > Up-SMetOH (9/16). Results obtained with Up-SMetOH against food spoiling Bacillus pumilus, Shigella spp. and Enterococcus gallinarum with minimum inhibitory concentrations (MICs) in 128-1024 μg/ml range seem to be promising. Up-SMetOH also exerted strong inhibition against Clavibacter michiganensis with a considerably low MIC (32 μg/ml). Ud-SMetOH and Up-LMetOH were also effective against C. michiganensis (MIC = 256 and 1024 μg/ml, respectively). Ud-SMetOH and Ud-RMetOH had also antimicrobial activity against Xanthomonas vesicatoria (MIC = 512 and 1024 μg/ml, respectively). Results presented here demonstrate high-antibacterial activity of U. pilulifera extracts and U. dioica seed extract against phytopathogens for the first time, and provide the most comprehensive data on the antibacterial activity screening of U. pilulifera against food-borne pathogens. Considering limitations in plant disease control, antibacterial activities of these extracts would be of agricultural importance.

  20. High-antibacterial activity of Urtica spp. seed extracts on food and plant pathogenic bacteria.

    PubMed

    Körpe, Didem Aksoy; İşerı, Özlem Darcansoy; Sahin, Feride Iffet; Cabi, Evren; Haberal, Mehmet

    2013-05-01

    The aim of this study was to comparatively evaluate antibacterial activities of methanol (MetOH) and aqueous (dw) leaf (L), root (R) and seed (S) extracts of Urtica dioica L. (Ud; stinging nettle) and Urtica pilulifera L. (Up; Roman nettle) on both food- and plant-borne pathogens, with total phenolic contents and DPPH radical scavenging activities (DRSA). MetOH extracts of leaves and roots of U. dioica had the highest DRSA. Extracts with high antibacterial activity were in the order Up-LMetOH (13/16) > Ud-SMetOH (11/16) > Up-SMetOH (9/16). Results obtained with Up-SMetOH against food spoiling Bacillus pumilus, Shigella spp. and Enterococcus gallinarum with minimum inhibitory concentrations (MICs) in 128-1024 μg/ml range seem to be promising. Up-SMetOH also exerted strong inhibition against Clavibacter michiganensis with a considerably low MIC (32 μg/ml). Ud-SMetOH and Up-LMetOH were also effective against C. michiganensis (MIC = 256 and 1024 μg/ml, respectively). Ud-SMetOH and Ud-RMetOH had also antimicrobial activity against Xanthomonas vesicatoria (MIC = 512 and 1024 μg/ml, respectively). Results presented here demonstrate high-antibacterial activity of U. pilulifera extracts and U. dioica seed extract against phytopathogens for the first time, and provide the most comprehensive data on the antibacterial activity screening of U. pilulifera against food-borne pathogens. Considering limitations in plant disease control, antibacterial activities of these extracts would be of agricultural importance. PMID:23067263

  1. Application of zinc chloride precipitation method for rapid isolation and concentration of infectious Pectobacterium spp. and Dickeya spp. lytic bacteriophages from surface water and plant and soil extracts.

    PubMed

    Czajkowski, Robert; Ozymko, Zofia; Lojkowska, Ewa

    2016-01-01

    This is the first report describing precipitation of bacteriophage particles with zinc chloride as a method of choice to isolate infectious lytic bacteriophages against Pectobacterium spp. and Dickeya spp. from environmental samples. The isolated bacteriophages are ready to use to study various (ecological) aspects of bacteria-bacteriophage interactions. The method comprises the well-known precipitation of phages from aqueous extracts of the test material by addition of ZnCl2, resuscitation of bacteriophage particles in Ringer's buffer to remove the ZnCl2 excess and a soft agar overlay assay with the host bacterium to isolate infectious individual phage plaques. The method requires neither an enrichment step nor other steps (e. g., PEG precipitation, ultrafiltration, or ultracentrifugation) commonly used in other procedures and results in isolation of active viable bacteriophage particles.

  2. Comparison of four commercial DNA extraction kits for the recovery of Bacillus spp. spore DNA from spiked powder samples.

    PubMed

    Mölsä, Markos; Kalin-Mänttäri, Laura; Tonteri, Elina; Hemmilä, Heidi; Nikkari, Simo

    2016-09-01

    Bacillus spp. include human pathogens such as Bacillus anthracis, the causative agent of anthrax and a biothreat agent. Bacillus spp. form spores that are physically highly resistant and may remain active over sample handling. We tested four commercial DNA extraction kits (QIAamp DNA Mini Kit, RTP Pathogen Kit, ZR Fungal/Bacterial DNA MiniPrep, and genesig Easy DNA/RNA Extraction kit) for sample inactivation and DNA recovery from two powders (icing sugar and potato flour) spiked with Bacillus thuringiensis spores. The DNA was analysed using a B. thuringiensis-specific real-time PCR assay. The detection limit was 3×10(1)CFU of spiked B. thuringiensis spores with the QIAamp DNA Mini, RTP Pathogen, and genesig Easy DNA/RNA Extraction kits, and 3×10(3)CFU with the ZR Fungal/Bacterial DNA MiniPrep kit. The results showed that manual extraction kits are effective and safe for fast and easy DNA extraction from powder samples even in field conditions. Adding a DNA filtration step to the extraction protocol ensures the removal of Bacillus spp. spores from DNA samples without affecting sensitivity. PMID:27435532

  3. Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae.

    PubMed

    Mogni, Virginia Y; Kahan, Mariano A; de Queiroz, Luciano Paganucci; Vesprini, José L; Ortiz, Juan Pablo A; Prado, Darién E

    2016-01-01

    The Anacardiaceae is an important and worldwide distributed family of ecological and socio-economic relevance. Notwithstanding that, molecular studies in this family are scarce and problematic because of the particularly high concentration of secondary metabolites-i.e. tannins and oleoresins-that are present in almost all tissues of the many members of the group, which complicate the purification and amplification of the DNA. The objective of this work was to improve an available DNA isolation method for Schinopsis spp. and other related Anacardiaceae, as well as the PCR protocols for DNA amplification of the chloroplast trnL-F, rps16 and ndhF and nuclear ITS-ETS fragments. The modifications proposed allowed the extraction of 70-120 µg of non-degraded genomic DNA per gram of dry tissue that resulted useful for PCR amplification. PCR reactions produced the expected fragments that could be directly sequenced. Sequence analyses of amplicons showed similarity with the corresponding Schinopsis accessions available at GenBank. The methodology presented here can be routinely applied for molecular studies of the group aimed to clarify not only aspects on the molecular biology but also the taxonomy and phylogeny of this fascinating group of vascular plants. PMID:27217992

  4. Anti-fish nodaviral activity of furan-2-yl acetate extracted from marine Streptomyces spp.

    PubMed

    Suthindhiran, K; Sarath Babu, V; Kannabiran, K; Ishaq Ahmed, V P; Sahul Hameed, A S

    2011-04-01

    The antiviral activity of furan-2-yl acetate (C₆H₆O₃) extracted from Streptomyces VITSDK1 spp. was studied in cultured Sahul Indian Grouper Eye (SIGE) cells infected with fish nodavirus (FNV). The nodavirus infection in the SIGE cells was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) and the antiviral activity of furan-2-yl acetate was assessed by cytopathic effect, as well as reduction in nodaviral titre (TCID₅₀ mL⁻¹, where TCID₅₀) is the 50% tissue culture infective dose) in the cultured SIGE cells under in vitro conditions. Furan-2-yl acetate (20 µg mL⁻¹) effectively inhibited the replication of the FNV-infected SIGE cell lines and the viral titre was reduced from 4.3 to 2.45 log TCID₅₀ mL⁻¹ on treatments. Furan-2-yl acetate (20 µg mL⁻¹)- treated SIGE cell survival was found to be 90%, as determined by methyl thiazol tetrazolium assay. The results of an immunofluorescent assay revealed a strong association between the viral capsid protein inhibition and a decline in viral replication. The results suggest that furan-2-yl acetate suppressed FNV replication in cultured fish cells, providing a potential approach for the control of nodaviral diseases in marine fishes. PMID:21462077

  5. Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae.

    PubMed

    Mogni, Virginia Y; Kahan, Mariano A; de Queiroz, Luciano Paganucci; Vesprini, José L; Ortiz, Juan Pablo A; Prado, Darién E

    2016-01-01

    The Anacardiaceae is an important and worldwide distributed family of ecological and socio-economic relevance. Notwithstanding that, molecular studies in this family are scarce and problematic because of the particularly high concentration of secondary metabolites-i.e. tannins and oleoresins-that are present in almost all tissues of the many members of the group, which complicate the purification and amplification of the DNA. The objective of this work was to improve an available DNA isolation method for Schinopsis spp. and other related Anacardiaceae, as well as the PCR protocols for DNA amplification of the chloroplast trnL-F, rps16 and ndhF and nuclear ITS-ETS fragments. The modifications proposed allowed the extraction of 70-120 µg of non-degraded genomic DNA per gram of dry tissue that resulted useful for PCR amplification. PCR reactions produced the expected fragments that could be directly sequenced. Sequence analyses of amplicons showed similarity with the corresponding Schinopsis accessions available at GenBank. The methodology presented here can be routinely applied for molecular studies of the group aimed to clarify not only aspects on the molecular biology but also the taxonomy and phylogeny of this fascinating group of vascular plants.

  6. Physiological effects of dietary fructans extracted from Agave tequilana Gto. and Dasylirion spp.

    PubMed

    Urías-Silvas, Judith E; Cani, Patrice D; Delmée, Evelyne; Neyrinck, Audrey; López, Mercedes G; Delzenne, Nathalie M

    2008-02-01

    Recent data reported that inulin-type fructans extracted from chicory roots regulate appetite and lipid/glucose metabolism, namely, by promoting glucagon-like peptide-1 (GLP-1) production in the colon. The Agave genus growing in different regions of Mexico also contains important amounts of original fructans, with interesting nutritional and technological properties, but only few data report their physiological effect when added in the diet. Therefore, we decided to evaluate in parallel the effect of supplementation with 10 % agave or chicory fructans on glucose and lipid metabolism in mice. Male C57Bl/6J mice were fed a standard (STD) diet or diet supplemented with Raftilose P95 (RAF), fructans from Agave tequilana Gto. (TEQ) or fructans from Dasylirion spp. (DAS) for 5 weeks. The body weight gain and food intake in mice fed fructans-containing diets were significantly lower than the ones of mice fed the STD diet, TEQ leading to the lowest value. Serum glucose and cholesterol were similarly lower in all fructans-fed groups than in the STD group and correlated to body weight gain. Only RAF led to a significant decrease in serum TAG. As previously shown for RAF, the supplementation with agave fructans (TEQ and DAS) induced a higher concentration of GLP-1 and its precursor, proglucagon mRNA, in the different colonic segments, thus suggesting that fermentable fructans from different botanical origin and chemical structure are able to promote the production of satietogenic/incretin peptides in the lower part of the gut, with promising effects on glucose metabolism, body weight and fat mass development.

  7. Fast protocol for extraction of DNA from Prosopis spp leaves (plant adapted to arid environment) without liquid nitrogen.

    PubMed

    Michel-López, C Y; González-Mendoza, D; Grimaldo-Juarez, O

    2013-09-27

    The extraction of high-quality genomic DNA from Prosopis spp for polymerase chain reaction (PCR) amplification is complicated, owing to the presence of a high percentage of secondary metabolites that bind to or co-precipitate with nucleic acids. In the present study, we report a modified sodium dodecyl sulfate/phenol protocol that eliminates the use of liquid nitrogen in the maceration process, β-mercaptoethanol in the buffer extraction, and the ethanol precipitation step. The A₂₆₀/A₂₈₀ absorbance ratios of the isolated DNA were approximately 2.0 to 1.9, suggesting that the DNA fraction was pure and can be used for further PCR analysis. The DNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder DNA extraction. Finally, this proposal represents an alternative fast, cheap, and effective method for the isolation of genomic DNA from fresh leaves of Prosopis spp, even in low-technology laboratories.

  8. Seasonal distribution of Legionella spp. and L. pneumophila in a river in Taiwan evaluated with culture-confirmed and direct DNA extraction methods

    NASA Astrophysics Data System (ADS)

    Tung, Min-Che; Chang, Tien-Yu; Hsu, Bing-Mu; Shen, Shu-Min; Huang, Jen-Te; Kao, Po-Min; Chiu, Yi-Chou; Fan, Cheng-Wei; Huang, Yu-Li

    2013-07-01

    In this study, we evaluated the presence and amount of Legionella in along a river in Taiwan, and the relations between seasonal distribution of Legionella spp. and geographic characteristics in the watershed were also evaluated. Water samples were pre-treated and analyzed with culture-confirmed and direct DNA extraction methods. For culture-confirmed method, water samples were cultivated through a series of selective media, and candidate colonies were confirmed by PCR. For direct DNA extraction method, direct DNA extraction was performed from pre-treated water samples. The DNA extracts were analyzed with PCR and DNA sequence analysis for species determination, quantitative PCR (qPCR) was performed to quantify Legionella concentration in the water sample. In all, 150 water samples were included in this study, with 73 (48.6%) water samples detected with Legionella spp., and 17 with L. pneumophila. Over 80% Legionella spp. detections were through direct DNA extraction method, but more than 80% L. pneumophila detections were through culture-confirmed method. While detection of Legionella spp. was done with two methods, positive results were found through only one method. Legionella spp. was detected in all seasons with detection rate ranging between 34.3-58.8% and seasonal average concentration from 1.9 × 102 to 7.1 × 103 CFU/L. Most of the L. pneumophila detections were from samples collected in fall (38.2%) and summer (6.0%), which also coincided with increased cases of Legionellosis reported through Center of Disease Control in Taiwan. The high prevalence and concentration of Legionella spp. and L. pneumophila in the surface waters should be further evaluated for potential health risks.

  9. Antigenotoxic Effect of Trametes spp. Extracts against DNA Damage on Human Peripheral White Blood Cells

    PubMed Central

    Knežević, Aleksandar; Živković, Lada; Stajić, Mirjana; Vukojević, Jelena; Milovanović, Ivan; Spremo-Potparević, Biljana

    2015-01-01

    Trametes species have been used for thousands of years in traditional and conventional medicine for the treatment of various types of diseases. The goal was to evaluate possible antigenotoxic effects of mycelium and basidiocarp extracts of selected Trametes species and to assess dependence on their antioxidant potential. Trametes versicolor, T. hirsuta, and T. gibbosa were the species studied. Antigenotoxic potentials of extracts were assessed on human peripheral white blood cells with basidiocarp and mycelium extracts of the species. The alkaline comet test was used for detection of DNA strand breaks and alkali-labile sites, as well as the extent of DNA migration. DPPH assay was used to estimate antioxidative properties of extracts. Fruiting body extracts of T. versicolor and T. gibbosa as well as T. hirsuta extracts, except that at 20.0 mg/mL, were not genotoxic agents. T. versicolor extract had at 5.0 mg/mL the greatest antigenotoxic effect in both pre- and posttreatment of leukocytes. The mycelium extracts of the three species had no genotoxic activity and significant antigenotoxic effect against H2O2-induced DNA damage, both in pre- and posttreatment. The results suggest that extracts of these three species could be considered as strong antigenotoxic agents able to stimulate genoprotective response of cells. PMID:26258163

  10. Antioxidant capacity and phenolic content in leaf extracts of tree spinach (Cnidoscolus spp.).

    PubMed

    Kuti, Joseph O; Konuru, Hima B

    2004-01-14

    Total phenolic content and antioxidant capacity of two tree spinach species (Cnidoscolus chayamansa McVaugh and C. aconitifolius Miller.) were determined in raw and cooked leaf extracts. Antioxidant capacity was assessed by the oxygen radical absorbance capacity (ORAC) assay, and flavonoid glycoside composition was quantified by HPLC and identified by GC. Total phenolics and antioxidant capacity were higher in raw than in cooked leaf extracts. The ORAC values were strongly correlated with total phenolic content (r = 0.926) in all leaf extracts. The major flavonoids isolated from the leaf extracts were kaempferol-3-O-glycosides and quercetin-3-O-glycosides. C. aconitifolius leaves contained more varieties of the flavonoid glycosides than C. chayamansa. Cooking reduced antioxidant activity and phenolic content and resulted in losses of some kaempferol glycoside and quercetin glycoside residues in leaf extracts. The results of this study indicate that tree spinach leaves are a rich source of natural antioxidants for foods. PMID:14709023

  11. Optimization of carvacrol, rosmarinic, oleanolic and ursolic acid extraction from oregano herbs (Origanum onites L., Origanum vulgare spp. hirtum and Origanum vulgare L.).

    PubMed

    Baranauskaitė, Justė; Jakštas, Valdas; Ivanauskas, Liudas; Kopustinskienė, Dalia M; Drakšienė, Gailutė; Masteikova, Ruta; Bernatonienė, Jurga

    2016-01-01

    The aim of our study was to increase the extraction efficiency of carvacrol, rosmarinic, oleanolic and ursolic acid from the different species of oregano herbs (Origanum onites L., Origanum vulgare spp. hirtum and Origanum vulgare L.). Various extraction methods (ultrasound-assisted, heat-reflux, continuous stirring, maceration, percolation) and extraction conditions (different solvent, material:solvent ratio, extraction temperature, extraction time) were used, and the active substances were determined by HPLC. The lowest content of carvacrol, rosmarinic, oleanolic and ursolic acid was obtained by percolation. During heat-reflux extraction, the content of active substances depended on the solvent used: ethanol/non-aqueous solvent (glycerol or propylene glycol) mixture was more effective compared with ethanol alone. The results showed that for each species of oregano the most optimal extraction method should be selected to maximize the content of biologically active substances in the extracts.

  12. Anticomplement activity of various solvent extracts from Korea local Artemisia spp.

    PubMed

    Moon, Hyung-In; Jung, Seil; Lee, Young-Choon; Lee, Jai-Heon

    2012-02-01

    The study evaluated the anticomplement activity from various solvent extracts of eight Artemisia plants (Artemisia capillaris Thunb., Artemisia fukudo Makino., Artemisia japonica Thunb., Artemisia montana (Nakai) Pamp., Artemisia keiskeana Miq., Artemisia rubripes Nakai., Artemisia stolonifera (Maxim.) Kom., and Artemisia sylvatica Max.) from South Korea on the classical pathway (CP). We have evaluated various organic solvent extract from eight Artemisia plants with regard to its anticomplement activity on the CP. A. rubripes and A. montana chloroform extracts showed inhibitory activity against complement system with 50% inhibitory concentrations (IC₅₀) values of 54.3 and 64.2 μg/mL. This is the first report of anticomplement activity from Artemisia plants.

  13. Suitability of Bifidobacterium spp. and Lactobacillus plantarum as probiotics intended for fruit juices containing citrus extracts.

    PubMed

    Bevilacqua, Antonio; Campaniello, Daniela; Corbo, Maria Rosaria; Maddalena, Lucia; Sinigaglia, Milena

    2013-11-01

    A strain of Lactobacillus plantarum and 4 strains of bifidobacteria were inoculated in apple juice and in a commercial beverage labeled as "red-fruit juice," containing citrus extracts as natural preservatives; the suitability of the probiotics was evaluated in relation to their resistance to 2 kinds of citrus extracts (biocitro and lemon extract), survival in juices at 4 and 37 °C, and inhibition of Zygosaccharomyces bailii. Cell count of L. plantarum and bifidobacteria over time was fitted through the Weibull equation, for the evaluation of the first reduction time (δ), death time, and microbiological shelf life (the break-point was set to 7 log cfu/mL). Bifidobacterium animalis subsp. lactis experienced the highest δ-value (23.21 d) and death time (96.59 d) in the red-fruit juice at 4 °C, whereas L. plantarum was the most promising strain in apple juice at 37 °C. Biocitro and lemon extract did not exert a biocidal effect toward probiotics; moreover, the probiotics controlled the growth of Z. bailii and the combination of L. plantarum with 40 ppm of biocitro reduced the level of the yeast after 18 d by 2 log cfu/mL.

  14. Toxin induction and protein extraction from Fusarium spp. cultures for proteomic studies.

    PubMed

    Pasquali, Matias; Giraud, Frédéric; Lasserre, Jean Paul; Planchon, Sebastien; Hoffmann, Lucien; Bohn, Torsten; Renaut, Jenny

    2010-01-01

    Fusaria are filamentous fungi able to produce different toxins. Fusarium mycotoxins such as deoxynivalenol, nivalenol, T2, zearelenone, fusaric acid, moniliformin, etc... have adverse effects on both human and animal health and some are considered as pathogenicity factors. Proteomic studies showed to be effective for deciphering toxin production mechanisms (Taylor et al., 2008) as well as for identifying potential pathogenic factors (Paper et al., 2007, Houterman et al., 2007) in Fusaria. It becomes therefore fundamental to establish reliable methods for comparing between proteomic studies in order to rely on true differences found in protein expression among experiments, strains and laboratories. The procedure that will be described should contribute to an increased level of standardization of proteomic procedures by two ways. The filmed protocol is used to increase the level of details that can be described precisely. Moreover, the availability of standardized procedures to process biological replicates should guarantee a higher robustness of data, taking into account also the human factor within the technical reproducibility of the extraction procedure. The protocol described requires 16 days for its completion: fourteen days for cultures and two days for protein extraction (figure 1). Briefly, Fusarium strains are grown on solid media for 4 days; they are then manually fragmented and transferred into a modified toxin inducing media (Jiao et al., 2008) for 10 days. Mycelium is collected by filtration through a Miracloth layer. Grinding is performed in a cold chamber. Different operators performed extraction replicates (n=3) in order to take into account the bias due to technical variations (figure 2). Extraction was based on a SDS/DTT buffer as described in Taylor et al. (2008) with slight modifications. Total protein extraction required a precipitation process of the proteins using Aceton/TCA/DTT buffer overnight and Acetone /DTT washing (figure 3a,3b

  15. Improved methods for capture, extraction, and quantitative assay of environmental DNA from Asian bigheaded carp (Hypophthalmichthys spp.).

    PubMed

    Turner, Cameron R; Miller, Derryl J; Coyne, Kathryn J; Corush, Joel

    2014-01-01

    Indirect, non-invasive detection of rare aquatic macrofauna using aqueous environmental DNA (eDNA) is a relatively new approach to population and biodiversity monitoring. As such, the sensitivity of monitoring results to different methods of eDNA capture, extraction, and detection is being investigated in many ecosystems and species. One of the first and largest conservation programs with eDNA-based monitoring as a central instrument focuses on Asian bigheaded carp (Hypophthalmichthys spp.), an invasive fish spreading toward the Laurentian Great Lakes. However, the standard eDNA methods of this program have not advanced since their development in 2010. We developed new, quantitative, and more cost-effective methods and tested them against the standard protocols. In laboratory testing, our new quantitative PCR (qPCR) assay for bigheaded carp eDNA was one to two orders of magnitude more sensitive than the existing endpoint PCR assays. When applied to eDNA samples from an experimental pond containing bigheaded carp, the qPCR assay produced a detection probability of 94.8% compared to 4.2% for the endpoint PCR assays. Also, the eDNA capture and extraction method we adapted from aquatic microbiology yielded five times more bigheaded carp eDNA from the experimental pond than the standard method, at a per sample cost over forty times lower. Our new, more sensitive assay provides a quantitative tool for eDNA-based monitoring of bigheaded carp, and the higher-yielding eDNA capture and extraction method we describe can be used for eDNA-based monitoring of any aquatic species.

  16. Improved Methods for Capture, Extraction, and Quantitative Assay of Environmental DNA from Asian Bigheaded Carp (Hypophthalmichthys spp.)

    PubMed Central

    Turner, Cameron R.; Miller, Derryl J.; Coyne, Kathryn J.; Corush, Joel

    2014-01-01

    Indirect, non-invasive detection of rare aquatic macrofauna using aqueous environmental DNA (eDNA) is a relatively new approach to population and biodiversity monitoring. As such, the sensitivity of monitoring results to different methods of eDNA capture, extraction, and detection is being investigated in many ecosystems and species. One of the first and largest conservation programs with eDNA-based monitoring as a central instrument focuses on Asian bigheaded carp (Hypophthalmichthys spp.), an invasive fish spreading toward the Laurentian Great Lakes. However, the standard eDNA methods of this program have not advanced since their development in 2010. We developed new, quantitative, and more cost-effective methods and tested them against the standard protocols. In laboratory testing, our new quantitative PCR (qPCR) assay for bigheaded carp eDNA was one to two orders of magnitude more sensitive than the existing endpoint PCR assays. When applied to eDNA samples from an experimental pond containing bigheaded carp, the qPCR assay produced a detection probability of 94.8% compared to 4.2% for the endpoint PCR assays. Also, the eDNA capture and extraction method we adapted from aquatic microbiology yielded five times more bigheaded carp eDNA from the experimental pond than the standard method, at a per sample cost over forty times lower. Our new, more sensitive assay provides a quantitative tool for eDNA-based monitoring of bigheaded carp, and the higher-yielding eDNA capture and extraction method we describe can be used for eDNA-based monitoring of any aquatic species. PMID:25474207

  17. Plant Extracts of Straw from Sugarcane (Saccharum spp.) in the Attenuation of Toxicity by Aluminum

    NASA Astrophysics Data System (ADS)

    Malvestiti, Jacqueline Ap.; Soares, Marcio Roberto; Casagrande, José Carlos

    2014-05-01

    Organic acids from decomposition of sugar cane straw are capable of interacting with elements of the soil solution, attenuating the toxicity by aluminum (Al) and promoting greater movement of cations in the soil profile. This research had as objective to analyze organic acids present in the straw of the sugarcane varieties RB855453, RB966928. The experiment was conducted under laboratory conditions. The experimental design used was the completely randomized, with five repetitions. The results showed that the analysis, chemical characterization and determination of water-soluble organic compounds of plant extracts (malic and acetic acid) was of great importance for the understanding of the development of the root system of sugarcane considering the soil management systems, since they provide information about the ability of the attenuation of the Al, exchangeable acidity of the soil and the mobility of basic cations to the soil sub layers. This study pointed out greater power of exchangeable cations transport throughout the soil profile, and Al neutralization phytotoxic by the vegetable extract of straw of RB867515 variety, because, besides highest content of basic cations and greater electric conductivity, the total concentration of organic acids was higher on the vegetable extract from the straw of this variety.

  18. Hydroxycinnamic Acid Derivatives Obtained from a Commercial Crataegus Extract and from Authentic Crataegus spp

    PubMed Central

    Kuczkowiak, Ulrich; Petereit, Frank; Nahrstedt, Adolf

    2014-01-01

    Abstract Eleven hydroxycinnamic acid derivatives were isolated from a 70% methanolic Crataegus extract (Crataegi folium cum flore) and partly verified and quantified for individual Crataegus species (C. laevigata, C. monogyna, C. nigra, C. pentagyna) by HPLC: 3-O-(E)-p-coumaroylquinic acid (1), 5-O-(E)-p-coumaroyl-quinic acid (2), 4-O-(E)-p-coumaroylquinic acid (3), 3-O-(E)-caffeoylquinic acid (4), 4-O-(E)-caffeoylquinic acid (5), 5-O-(E)-caffeoylquinic acid (6), 3,5-di-O-(E)-caffeoylquinic acid (7), 4,5-di-O-(E)-caffeoylquinic acid (8), (-)-2-O-(E)-caffeoyl-L-threonic acid (9), (-)-4-O-(E)-caffeoyl-L-threonic acid (10), and (-)-4-O-(E)-p-coumaroyl-L-threonic acid (11). Further, (-)-2-O-(E)-caffeoyl-D-malic acid (12) was isolated from C. submollis and also identified for C. pentagyna and C. nigra by co-chromatography. The isolates 10 and 11 were not found in the authentic fresh specimen, indicating that they may be formed during extraction by acyl migration from the 2-O-acylderivatives. Also, 9 and 11 are described here for the first time. All structures were assigned on the basis of their spectroscopic data (1H-, 13C-NMR, MS, optical rotation). PMID:26171328

  19. Genotoxicity in Oreochromis niloticus (Cichlidae) induced by Microcystis spp bloom extract containing microcystins.

    PubMed

    da Silva, R R Pavan; Pires, O R; Grisolia, C K

    2011-09-01

    Studies of genotoxicity in fish caused by cyanobacterial extracts containing microcystins (MCs) can be useful in determining their carcinogenic risk due to a genotoxic mechanism. An extract of cyanobacterial Microcystis ssp, containing MC-LR and -LA from a bloom collected in a eutrophic lake, showed genotoxicity to Oreochromis niloticus. DNA damage (comet assay) was significantly induced in peripheral erythrocytes with both tested concentrations of 6.90 μg kg(-1) bw and 13.80 μg kg(-1) bw through intraperitoneal injection (ip). There was no micronucleus induction after ip injection at concentrations of 6.90 μg kg(-1) bw and 13.80 μg kg(-1) bw. Body exposure resulted in micronucleus induction and DNA damage only at the highest tested concentrations of 103.72 μg L(-1). Thus, comet assay and ip injection revealed the highest levels of the genotoxicity of MCs. Apoptosis-necrosis test carried out at concentrations of 6.90 μg kg(-1) bw and 13.80 μg kg(-1) bw revealed that at low concentrations more apoptosis than necrosis occurred. At higher concentrations more necrosis than apoptosis occurred. PMID:21704053

  20. Arrabidaea chica Hexanic Extract Induces Mitochondrion Damage and Peptidase Inhibition on Leishmania spp.

    PubMed Central

    Rodrigues, Igor A.; Azevedo, Mariana M. B.; Chaves, Francisco C. M.; Alviano, Celuta S.; Alviano, Daniela S.; Vermelho, Alane B.

    2014-01-01

    Currently available leishmaniasis treatments are limited due to severe side effects. Arrabidaea chica is a medicinal plant used in Brazil against several diseases. In this study, we investigated the effects of 5 fractions obtained from the crude hexanic extract of A. chica against Leishmania amazonensis and L. infantum, as well as on the interaction of these parasites with host cells. Promastigotes were treated with several concentrations of the fractions obtained from A. chica for determination of their minimum inhibitory concentration (MIC). In addition, the effect of the most active fraction (B2) on parasite's ultrastructure was analyzed by transmission electron microscopy. To evaluate the inhibitory activity of B2 fraction on Leishmania peptidases, parasites lysates were treated with the inhibitory and subinhibitory concentrations of the B2 fraction. The minimum inhibitory concentration of B2 fraction was 37.2 and 18.6 μg/mL for L. amazonensis and L. infantum, respectively. Important ultrastructural alterations as mitochondrial swelling with loss of matrix content and the presence of vesicles inside this organelle were observed in treated parasites. Moreover, B2 fraction was able to completely inhibit the peptidase activity of promastigotes at pH 5.5. The results presented here further support the use of A. chica as an interesting source of antileishmanial agents. PMID:24818162

  1. Antifungal activity of the ethanolic extracts of Punica granatum L. and evaluation of the morphological and structural modifications of its compounds upon the cells of Candida spp.

    PubMed

    Anibal, Paula Cristina; Peixoto, Iza Teixeira Alves; Foglio, Mary Ann; Höfling, José Francisco

    2013-01-01

    Ethanolic crude extracts prepared from the arils and seeds, pericarp, peels and from the whole fruit of Punica granatum, known as pomegranate, had their antifungal activity tested against Candida spp. The ethanolic crude extracts were analyzed by Mass Spectrometry and yielded many compounds such as punicalagin and galladydilacton. The extracts from the pericarp and peel showed activity against Candida spp., with MICs of 125 μg/mL. The effect of pericarp and peel extracts upon the morphological and structure of C. albicans and C. krusei were examined by scanning and transmission electron microscopy, with the visualization of an irregular membrane and hyphae, formation of vacuoles and thickening of the cell wall. The data obtained revealed potential antimicrobial activity against yeasts cells of the Candida genus, and the bioactive compounds could be responsible for changes in cell morphology and structure. The data obtained open new perspectives for future research in continuation to this study, where information such as determination of the site of action of the compounds could contribute to an alternative therapy against these organisms.

  2. Antifungal activity of the ethanolic extracts of Punica granatum L. and evaluation of the morphological and structural modifications of its compounds upon the cells of Candida spp

    PubMed Central

    Anibal, Paula Cristina; Peixoto, Iza Teixeira Alves; Foglio, Mary Ann; Höfling, José Francisco

    2013-01-01

    Ethanolic crude extracts prepared from the arils and seeds, pericarp, peels and from the whole fruit of Punica granatum, known as pomegranate, had their antifungal activity tested against Candida spp. The ethanolic crude extracts were analyzed by Mass Spectrometry and yielded many compounds such as punicalagin and galladydilacton. The extracts from the pericarp and peel showed activity against Candida spp., with MICs of 125 μg/mL. The effect of pericarp and peel extracts upon the morphological and structure of C. albicans and C. krusei were examined by scanning and transmission electron microscopy, with the visualization of an irregular membrane and hyphae, formation of vacuoles and thickening of the cell wall. The data obtained revealed potential antimicrobial activity against yeasts cells of the Candida genus, and the bioactive compounds could be responsible for changes in cell morphology and structure. The data obtained open new perspectives for future research in continuation to this study, where information such as determination of the site of action of the compounds could contribute to an alternative therapy against these organisms. PMID:24516425

  3. Sideritis spp. Extracts Enhance Memory and Learning in Alzheimer’s β-Amyloidosis Mouse Models and Aged C57Bl/6 Mice

    PubMed Central

    Hofrichter, Jacqueline; Krohn, Markus; Schumacher, Toni; Lange, Cathleen; Feistel, Bjöorn; Walbroel, Bernd; Pahnke, Jens

    2016-01-01

    Nowadays, Alzheimer’s disease is the most prevalent epiphenomenon of the aging population. Although soluble amyloid-β (Aβ) species (monomers, oligomers) are recognized triggers of the disease, no therapeutic approach is able to stop it. Herbal medicines are used to treat different diseases in many regions of the world. On the Balkan Peninsula, at the eastern Mediterranean Sea, and adjacent regions, Sideritis species are used as traditional medicine to prevent age-related problems in elderly. To evaluate this traditional knowledge in controlled experiments, we tested extracts of two commonly used Sideritis species, Sideritis euboea and Sideritis scardica, with regard to their effects on cognition in APP-transgenic and aged, non-transgenic C57Bl/6 mice. Additionally, histomorphological and biochemical changes associated with Aβ deposition and treatment were assessed. We found that daily oral treatment with Sideritis spp. extracts highly enhanced cognition in aged, non-transgenic as well as in APP-transgenic mice, an effect that was even more pronounced when extracts of both species were applied in combination. The treatment strongly reduced Aβ42 load in APP-transgenic mice, accompanied by increased phagocytic activity of microglia, and increased expression of the α-secretase ADAM10. Moreover, the treatment was able to fully rescue neuronal loss of APP-transgenic mice to normal levels as seen in non-transgenic controls. Having the traditional knowledge in mind, our results imply that treatment with Sideritis spp. extracts might be a potent, well-tolerated option for treating symptoms of cognitive impairment in elderly and with regard to Alzheimer’s disease by affecting its most prominent hallmarks: Aβ pathology and cognitive decline. PMID:27258424

  4. SIMPLE METHOD FOR THE EXTRACTION OF PHOTOPIGMENTS AND MYCOSPORINE-LIKE AMINO ACIDS (MAAS) FROM SYMBIODINIUM SPP.

    EPA Science Inventory

    Numerous extraction methods have been developed and used in the quantitation of both photopigments and mycosporine amino acids (MAAs) found in Symbiodinium sp. and zooanthellate metazoans. We have development of a simple, mild extraction procedure using methanol, which when coupl...

  5. Evaluation of an IgE ELISA with Culicoides spp. extracts and recombinant salivary antigens for diagnosis of insect bite hypersensitivity in Warmblood horses.

    PubMed

    Peeters, L M; Janssens, S; Goddeeris, B M; De Keyser, K; Wilson, A D; Kaufmann, C; Schaffartzik, A; Marti, E; Buys, N

    2013-10-01

    Insect bite hypersensitivity (IBH) in horses represents an immunoglobulin E (IgE)-mediated hypersensitivity to salivary antigens from biting midges (Culicoides spp.). The aim of this study was to evaluate and compare the performances of IgE ELISAs using recombinant Culicoides spp. Obsoletus group salivary gland antigens or crude whole body extracts ('ObsWBE'), C. nubeculosus recombinant proteins (Culn1, 3, 4, 5, 7, 8 and 10) and Obsoletus group recombinant proteins (Culo1 and 2). IgE levels were measured in plasma of 343 Warmblood horses classified as IBH-affected (n=167) and IBH-unaffected (n=176) according to the owners' descriptions. IBH-affected horses were subdivided based on the severity of their clinical signs at sampling and whether or not their IBH history was considered to be classical. The accuracies of the tests increased when clinical signs at sampling were more pronounced or when the IBH history could be considered as classical. A combination of IgE levels against the three best performing Culicoides spp. recombinant proteins (Culn4, Culo1 and Culo2) and ObsWBE resulted in the best performing test. When IBH-affected horses showing a classical history of the disease and severe clinical signs were compared with IBH-unaffected horses, the Youden's index at the optimal cut-off for the three tests in combination was 0.67. This optimal cut-off had a sensitivity of 70%, a specificity of 97% and a total accuracy of 92%. The performance of the IgE ELISA was affected by the severity of IBH clinical signs at sampling and was improved when IgE levels against several recombinant proteins were combined.

  6. In vivo antiprotozoan effects of garlic (Allium sativum) and ginger (Zingiber officinale) extracts on experimentally infected mice with Blastocystis spp.

    PubMed

    Abdel-Hafeez, Ekhlas H; Ahmad, Azza K; Kamal, Amany M; Abdellatif, Manal Z M; Abdelgelil, Noha H

    2015-09-01

    Controversy surrounding the pathogenic role of Blastocystis spp. in humans and lack of well-established diagnostic criteria led to debates concerning the treatment for that organism. Furthermore, some strains develop resistance against the recommended drugs. Thus, using natural medicine has many positive aspects to address these points. In an earlier study, we addressed in vitro effect of garlic and ginger on Blastocystis spp. isolates as an alternative treatment. Accordingly, this study was conducted to evaluate in vivo activities of these two herbs on mice infected with Blastocystis spp. Antiprotozoan activities were determined by monitoring Blastocystis shedding in stools and histopathological changes of the intestine of infected mice. Additionally, assessment of the antioxidant effect (via measuring the level of malondialdehyde (MDA) production) of these herbs on the treated groups of mice was done. Also, their effects on nitric oxide (NO) production were assessed. In this work, treatment of infected mice with garlic, ginger, and nitazoxanide (NTZ) reduced the shedding of cysts significantly compared to the infected untreated group, P value ≤0.001, 0.0001, and 0.0003, respectively. As well, histopathological examination revealed that Blastocystis was frequently observed within the lumen, at the tip of the epithelium, and/ or infiltrated in an enterocyte in the infected group without treatment compared to that of the infected treated ones. Furthermore, mice infected with Blastocystis exhibited increased levels of NO (440.09 ± 3.7 vs. 276.66 ± 0.8, P ≤ 0.001) and MDA production (106.19 ± 0.43 vs. 63.06 ± 0.45, P ≤ 0.0004) compared to that of the uninfected controls. Treatment of infected mice with garlic, ginger, and NTZ reduced NO levels to 54.41 ± 1.2, 47.70 ± 1.2, and 37.43 ± 0.98 and MDA levels to 22.38 ± 0.17, 63.34 ± 3.89, and 66.76 ± 9.1, respectively. We conclude that using ginger and garlic for treatment of blastocystosis is beneficial.

  7. Evaluation of different protein extraction methods for banana (Musa spp.) root proteome analysis by two-dimensional electrophoresis.

    PubMed

    Vaganan, M Mayil; Sarumathi, S; Nandakumar, A; Ravi, I; Mustaffa, M M

    2015-02-01

    Four protocols viz., the trichloroacetic acid-acetone (TCA), phenol-ammonium acetate (PAA), phenol/SDS-ammonium acetate (PSA) and trisbase-acetone (TBA) were evaluated with modifications for protein extraction from banana (Grand Naine) roots, considered as recalcitrant tissues for proteomic analysis. The two-dimensional electrophoresis (2-DE) separated proteins were compared based on protein yield, number of resolved proteins, sum of spot quantity, average spot intensity and proteins resolved in 4-7 pI range. The PAA protocol yielded more proteins (0.89 mg/g of tissues) and protein spots (584) in 2-DE gel than TCA and other protocols. Also, the PAA protocol was superior in terms of sum of total spot quantity and average spot intensity than TCA and other protocols, suggesting phenol as extractant and ammonium acetate as precipitant of proteins were the most suitable for banana rooteomics analysis by 2-DE. In addition, 1:3 ratios of root tissue to extraction buffer and overnight protein precipitation were most efficient to obtain maximum protein yield.

  8. Ethanolic extracts of Brassica campestris spp. rapa roots prevent high-fat diet-induced obesity via beta(3)-adrenergic regulation of white adipocyte lipolytic activity.

    PubMed

    An, Sojin; Han, Jang-Il; Kim, Min-Jung; Park, Ji-Seon; Han, Jong-Min; Baek, Nam-In; Chung, Hae-Gon; Choi, Myung-Sook; Lee, Kyung-Tae; Jeong, Tae-Sook

    2010-04-01

    The influence of ethanolic extracts of Brassica campestris spp. rapa roots (EBR) on obesity was examined in imprinting control region (ICR) mice fed a high-fat diet (HFD) and in 3T3-L1 adipocytes. The ICR mice used were divided into regular diet, HFD, EBR (50 mg/kg/day EBR administered orally), and orlistat (10 mg/kg/day orlistat administered orally) groups. The molecular mechanism of the anti-obesity effect of EBR was investigated in 3T3-L1 adipocytes as well as in HFD-fed ICR mice. In the obese mouse model, both weight gain and epididymal fat accumulation were highly suppressed by the daily oral administration of 50 mg/kg EBR for 8 weeks, whereas the overall amount of food intake was not affected. EBR treatment induced the expression in white adipocytes of lipolysis-related genes, including beta(3)-adrenergic receptor (beta(3)-AR), hormone-sensitive lipase (HSL), adipose triglyceride lipase, and uncoupling protein 2. Furthermore, the activation of cyclic AMP-dependent protein kinase, HSL, and extracellular signal-regulated kinase was induced in EBR-treated 3T3-L1 cells. The lipolytic effect of EBR involved beta(3)-AR modulation, as inferred from the inhibition by the beta(3)-AR antagonist propranolol. These results suggest that EBR may have potential as a safe and effective anti-obesity agent via the inhibition of adipocyte lipid accumulation and the stimulation of beta(3)-AR-dependent lipolysis.

  9. Metabolomic assessment with CE-MS of the nutraceutical effect of Cystoseira spp extracts in an animal model.

    PubMed

    Moraes, Edgar P; Rupérez, Francisco Javier; Plaza, Merichel; Herrero, Miguel; Barbas, Coral

    2011-08-01

    There is a need of scientific evidence of claimed nutraceutical effects, but also there is a social movement towards the use of natural products and among them algae are seen as rich resources. Within this scenario, the development of methodology for rapid and reliable assessment of markers of efficiency and security of these extracts is necessary. The rat treated with streptozotocin has been proposed as the most appropriate model of systemic oxidative stress for studying antioxidant therapies. Cystoseira is a brown alga containing fucoxanthin and other carothenes whose pressure-assisted extracts were assayed to discover a possible beneficial effect on complications related to diabetes evolution in an acute but short-term model. Urine was selected as the sample and CE-TOF-MS as the analytical technique to obtain the fingerprints in a non-target metabolomic approach. Multivariate data analysis revealed a good clustering of the groups and permitted the putative assignment of compounds statistically significant in the classification. Interestingly a group of compounds associated to lysine glycation and cleavage from proteins was found to be increased in diabetic animals receiving vehicle as compared to control animals receiving vehicle (N6,N6,N6-trimethyl-L-lysine, N-methylnicotinamide, galactosylhydroxylysine, L-carnitine, N6-acetyl-N6-hydroxylysine, fructose-lysine, pipecolic acid, urocanic acid, amino-isobutanoate, formylisoglutamine. Fructoselysine significantly decreased after the treatment changing from a 24% increase to a 19% decrease. CE-MS fingerprinting of urine has provided a group of compounds different to those detected with other techniques and therefore proves the necessity of a cross-platform analysis to obtain a broad view of biological samples. PMID:21792987

  10. Comparative assay of glutathione S-transferase (GSTs) activity of excretory-secretory materials and somatic extract of Fasciola spp parasites.

    PubMed

    Alirahmi, Heshmatollah; Farahnak, Ali; Golmohamadi, Taghi; Esharghian, Mohammad Reza

    2010-01-01

    Fascioliasis is a worldwide parasitic disease in human and domestic animals. The causative agents of fascioliasis are Fasciola hepatica and Fasciola gigantica. In the recent years, fasciola resistance to drugs has been reported in the many of publications. Fasciola spp has detoxification system including GST enzyme which may be responsible for its resistance. Therefore , the aim of the study was to assay of GST enzyme activity in fasciola parasites. Fasciola gigantica and Fasciola hepatica helminths were collected from abattoir as a live and cultured in buffer media for 4 h at 37 °C. Excretory-Secretory products were collected and stored in -80◦C. F. gigantica and Fasciola hepatica were homogenized with homogenizing buffer in a glass homogenizer to prepare of somatic extract. Suspension was then centrifuged and supernatant was stored at -80°C. In order to assay the enzyme activity, excretory-secretory and somatic extracts in the form of cocktails (potassium phosphate buffer, reduced glutathione and 1-chloro-2,4-dinitrobenzene substrates) were prepared and their absorbance recorded for 5 minutes at 340 nm. The total and specific GST activity of F. gigantica somatic and ES products were obtained as 2916.00, 272.01 micromole/minute and 1.33, 1.70 micromole/minute/mg protein, respectively. Fasciola hepatica also showed 2705.00, 276.86 micromole/minute and 1.33, 1.52 micromole/minute/mg protein, respectively. These results are important for analysis of parasite survival / resistance to drugs which use for treatment of fascioliasis.

  11. Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp.

    PubMed

    Paulos, Silvia; Mateo, Marta; de Lucio, Aida; Hernández-de Mingo, Marta; Bailo, Begoña; Saugar, José M; Cardona, Guillermo A; Fuentes, Isabel; Mateo, María; Carmena, David

    2016-08-01

    High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n=29), giardiasis (n=47) and amoebiasis by Entamoeba histolytica (n=3) or E. dispar (n=10) and apparently healthy subjects (n=24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56°C were proven more efficient for the release of DNA from Cryptosporidium oocysts.

  12. Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp.

    PubMed

    Paulos, Silvia; Mateo, Marta; de Lucio, Aida; Hernández-de Mingo, Marta; Bailo, Begoña; Saugar, José M; Cardona, Guillermo A; Fuentes, Isabel; Mateo, María; Carmena, David

    2016-08-01

    High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n=29), giardiasis (n=47) and amoebiasis by Entamoeba histolytica (n=3) or E. dispar (n=10) and apparently healthy subjects (n=24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56°C were proven more efficient for the release of DNA from Cryptosporidium oocysts. PMID:27241828

  13. In vitro activity of essential oils extracted from plants used as spices against fluconazole-resistant and fluconazole-susceptible Candida spp.

    PubMed

    Pozzatti, Patrícia; Scheid, Liliane Alves; Spader, Tatiana Borba; Atayde, Margareth Linde; Santurio, Janio Morais; Alves, Sydney Hartz

    2008-11-01

    In the present study, the antifungal activity of selected essential oils obtained from plants used as spices was evaluated against both fluconazole-resistant and fluconazole-susceptible Candida spp. The Candida species studied were Candida albicans, Candida dubliniensis, Candida tropicalis, Candida glabrata, and Candida krusei. For comparison purposes, they were arranged in groups as C. albicans, C. dubliniensis, and Candida non-albicans. The essential oils were obtained from Cinnamomum zeylanicum Breyn, Lippia graveolens HBK, Ocimum basilicum L., Origanum vulgare L., Rosmarinus officinalis L., Salvia officinalis L., Thymus vulgaris L., and Zingiber officinale. The susceptibility tests were based on the M27-A2 methodology. The chemical composition of the essential oils was obtained by gas chromatography-mass spectroscopy and by retention indices. The results showed that cinnamon, Mexican oregano, oregano, thyme, and ginger essential oils have different levels of antifungal activity. Oregano and ginger essential oils were found to be the most and the least efficient, respectively. The main finding was that the susceptibilities of fluconazole-resistant C. albicans, C. dubliniensis, and Candida non-albicans to Mexican oregano, oregano, thyme, and ginger essential oils were higher than those of the fluconazole-susceptible yeasts (P<0.05). In contrast, fluconazole-resistant C. albicans and Candida non-albicans were less susceptible to cinnamon essential oil than their fluconazole-susceptible counterparts (P<0.05). A relationship between the yeasts' susceptibilities and the chemical composition of the essential oils studied was apparent when these 2 parameters were compared. Finally, basil, rosemary, and sage essential oils did not show antifungal activity against Candida isolates at the tested concentrations. PMID:18997851

  14. Peperomia pellucida leaf extract as immunostimulator in controlling motile aeromonad septicemia due to Aeromonas hydrophila in red hybrid tilapia, Oreochromis spp. farming

    PubMed Central

    Lee, S. W.; Sim, K. Y.; Wendy, W.; Zulhisyam, A. K.

    2016-01-01

    Aim: This study was revealed the potential of Peperomia pellucida leaf extract as an immunostimulator agent in controlling motile aeromonad septicemia due to Aeromonas hydrophila in red hybrid tilapia, Oreochromis sp. Materials and Methods: In the present study, minimum inhibitory concentration (MIC) of P. pellucida leaf extract against A. hydrophila was determined through two-fold microbroth dilution method. The plant extract was screening for its active compound using a gas chromatograph mass spectrometer, and the effectiveness of P. pellucida leaf extract as an immunostimulator agent was evaluated. The experimental fish were fed with medicated feed at three different concentrations (25 mg/kg, PP-25; 50 mg/kg, PP-50; and 100 mg/kg, PP-100) of P. pellucida leaf extract for 1 week before they were intraperitoneally exposed to A. hydrophila. Enzyme-linked immunosorbent assay was carried out to determine the value of antibody response to A. hydrophila in fish from a group of fish that received medicated feed, and the percentage of total cumulative mortality of the experimental fish were observed at the end of the experiment. Results: The results showed that the major bioactive compound is phytol (40%), and the MIC value was 31.5 mg/L. The value of antibody response to A. hydrophila in fish from a group of fish which received medicated feed (PP-25, 0.128±0.014 optical density [OD]; PP-50, 0.132±0.003 OD; and PP-100, 0.171±0.02 OD) was found significantly higher (p<0.05) compared to fish did not receive medicated feed (0.00 OD). Whereas, percentage cumulative mortality of fish from all groups of fish received medicated feed (PP-25, 18.0±3.2%; PP-50, 18.2±2.8%; and PP-100, 17.7±1.8%) were found significantly lower (p<0.05) compared to a group of fish did not receive medicated feed (83.2±1.4%). Conclusion: The findings of the present study indicated the huge potential of P. pellucida leaf extract as natural immunostimulator agent for aquaculture uses. PMID

  15. Commercial Honeybush (Cyclopia spp.) Tea Extract Inhibits Osteoclast Formation and Bone Resorption in RAW264.7 Murine Macrophages-An in vitro Study.

    PubMed

    Visagie, Amcois; Kasonga, Abe; Deepak, Vishwa; Moosa, Shaakirah; Marais, Sumari; Kruger, Marlena C; Coetzee, Magdalena

    2015-10-28

    Honeybush tea, a sweet tasting caffeine-free tea that is indigenous to South Africa, is rich in bioactive compounds that may have beneficial health effects. Bone remodeling is a physiological process that involves the synthesis of bone matrix by osteoblasts and resorption of bone by osteoclasts. When resorption exceeds formation, bone remodeling can be disrupted resulting in bone diseases such as osteoporosis. Osteoclasts are multinucleated cells derived from hematopoietic precursors of monocytic lineage. These precursors fuse and differentiate into mature osteoclasts in the presence of receptor activator of NF-kB ligand (RANKL), produced by osteoblasts. In this study, the in vitro effects of an aqueous extract of fermented honeybush tea were examined on osteoclast formation and bone resorption in RAW264.7 murine macrophages. We found that commercial honeybush tea extract inhibited osteoclast formation and TRAP activity which was accompanied by reduced bone resorption and disruption of characteristic cytoskeletal elements of mature osteoclasts without cytotoxicity. Furthermore, honeybush tea extract decreased expression of key osteoclast specific genes, matrix metalloproteinase-9 (MMP-9), tartrate resistant acid phosphatase (TRAP) and cathepsin K. This study demonstrates for the first time that honeybush tea may have potential anti-osteoclastogenic effects and therefore should be further explored for its beneficial effects on bone.

  16. Commercial Honeybush (Cyclopia spp.) Tea Extract Inhibits Osteoclast Formation and Bone Resorption in RAW264.7 Murine Macrophages—An in vitro Study

    PubMed Central

    Visagie, Amcois; Kasonga, Abe; Deepak, Vishwa; Moosa, Shaakirah; Marais, Sumari; Kruger, Marlena C.; Coetzee, Magdalena

    2015-01-01

    Honeybush tea, a sweet tasting caffeine-free tea that is indigenous to South Africa, is rich in bioactive compounds that may have beneficial health effects. Bone remodeling is a physiological process that involves the synthesis of bone matrix by osteoblasts and resorption of bone by osteoclasts. When resorption exceeds formation, bone remodeling can be disrupted resulting in bone diseases such as osteoporosis. Osteoclasts are multinucleated cells derived from hematopoietic precursors of monocytic lineage. These precursors fuse and differentiate into mature osteoclasts in the presence of receptor activator of NF-kB ligand (RANKL), produced by osteoblasts. In this study, the in vitro effects of an aqueous extract of fermented honeybush tea were examined on osteoclast formation and bone resorption in RAW264.7 murine macrophages. We found that commercial honeybush tea extract inhibited osteoclast formation and TRAP activity which was accompanied by reduced bone resorption and disruption of characteristic cytoskeletal elements of mature osteoclasts without cytotoxicity. Furthermore, honeybush tea extract decreased expression of key osteoclast specific genes, matrix metalloproteinase-9 (MMP-9), tartrate resistant acid phosphatase (TRAP) and cathepsin K. This study demonstrates for the first time that honeybush tea may have potential anti-osteoclastogenic effects and therefore should be further explored for its beneficial effects on bone. PMID:26516894

  17. Suppression of Pythium spp. by Trichoderma spp. during germination of tomato seeds in soilless growing media.

    PubMed

    Aerts, R; De Schutter, B; Rombouts, L

    2002-01-01

    In the Flemish horticulture Pythium spp. is an important pathogen of tomato plants (Lycopersicon esculenthum) in soilless growing media. Therefore some experiments were conducted to evaluate the possibility of decreasing the damage caused by Pythium spp. by Trichoderma spp. In a tray with several growing media, a suspension of Trichoderma conidia (10(6)/ml growing medium) was applied two weeks before sowing. On some objects, a compost extract (Biostimulus) was added. The growing media used in the experiment were rockwool, recycled rockwool and recycled coconut fibre. After sowing, the trays were covered with perlite. Three isolates of Trichoderma spp.: T. asperellum (Biofungus), T. harzianum (Tri 003) and Trichoderma sp. (KHK) and two isolates of Pythium spp.: P. ultimum (MUCL) en P. aphanidermatum (HRI, UK) were used. Propamocarb was used as a chemical standard. The use of coconut fibre growing medium resulted in a higher percentage (36%) of germination than the rockwool media when only Pythium spp. was used. The presence of the spontaneous developing microflora in the coconut fibre medium gave probably also a suppression of Pythium spp. For that reason the results of the suppression by Trichoderma spp. are not easy to explain and very variable on the different objects. Pythium ultimum was more suppressed than P. aphanidermatum on all the growing media and the application of all the Trichoderma isolates increased the germination percentage of tomato seeds. T. asperellum (Biofungus) gave on rockwool also a good result for the suppression of P. aphanidermatum (increasing of germination with 48%). This effect was comparable with the propamocarb treatment (48%). T. harzianum (Tri 003) gave a small suppression (22%) and Trichoderma sp. (KHK) gave almost no suppression of P. aphanidermatum (7%). When less Trichoderma conidia were applied the germination percentage decreased. The adding of a compost extract (Biostimulus) had no influence on the results. This experiment

  18. Isolation and Antimicrobial Testing of Aeromonas spp., Citrobacter spp., Cronobacter spp., Enterobacter spp., Escherichia spp., Klebsiella spp., and Trabulsiella spp. from the Gallbladder of Pigs.

    PubMed

    Evangelopoulou, Grammato; Filioussis, Georgios; Kritas, Spyridon; Kantere, Maria; Burriel, Angeliki R

    2015-01-01

    The presence of Gram-negative bacteria species, other than Salmonella spp., in the gallbladder of pigs was examined. Isolated Gram-negative bacteria were assigned to species using the Microgen™ GnA+B-ID Systems. Of the 64 isolated strains 43 were identified as Escherichia coli, seven as Enterobacter spp., three each as Klebsiella spp., Citrobacterfreundii, Aeromonas hydrophila and Cronobacter sakazakii and one each as Escherichiafergusonii and Trabulsiella guamensis. Their antibiograms showed very high resistance to ampicillin, amoxicillin, tetracycline, chloramphenicol and sulfamethoxazole/trimethoprim. It was concluded that the pigs' gallbladder is a reservoir of potentially pathogenic Gram-negative bacteria for pork consumers.

  19. Occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system, Paraná, Southern Brazil.

    PubMed

    Almeida, Jonatas Campos; Martins, Felippe Danyel Cardoso; Ferreira Neto, José Maurício; Santos, Maíra Moreira Dos; Garcia, João Luis; Navarro, Italmar Teodorico; Kuroda, Emília Kiyomi; Freire, Roberta Lemos

    2015-01-01

    The purpose of this study was to investigate the occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system. Samples of raw and treated water were collected and concentrated using the membrane filtration technique. Direct Immunofluorescence Test was performed on the samples. DNA extraction using a commercial kit was performed and the DNA extracted was submitted to a nested-PCR reaction (n-PCR) and sequencing. In the immunofluorescence, 2/24 (8.33%) samples of raw water were positive for Giardia spp.. In n-PCR and sequencing, 2/24 (8.33%) samples of raw water were positive for Giardia spp., and 2/24 (8.33%) samples were positive for Cryptosporidium spp.. The sequencing showed Cryptosporidium parvum and Giardia duodenalis DNA. In raw water, there was moderate correlation among turbidity, color and Cryptosporidium spp. and between turbidity and Giardia spp.. The presence of these protozoans in the water indicates the need for monitoring for water-treatment companies. PMID:26291147

  20. Bartonella spp. in Bats, Guatemala.

    PubMed

    Bai, Ying; Kosoy, Michael; Recuenco, Sergio; Alvarez, Danilo; Moran, David; Turmelle, Amy; Ellison, James; Garcia, Daniel L; Estevez, Alejandra; Lindblade, Kim; Rupprecht, Charles

    2011-07-01

    To better understand the role of bats as reservoirs of Bartonella spp., we estimated Bartonella spp. prevalence and genetic diversity in bats in Guatemala during 2009. We found prevalence of 33% and identified 21 genetic variants of 13 phylogroups. Vampire bat-associated Bartonella spp. may cause undiagnosed illnesses in humans.

  1. Bartonella spp. in Bats, Guatemala.

    PubMed

    Bai, Ying; Kosoy, Michael; Recuenco, Sergio; Alvarez, Danilo; Moran, David; Turmelle, Amy; Ellison, James; Garcia, Daniel L; Estevez, Alejandra; Lindblade, Kim; Rupprecht, Charles

    2011-07-01

    To better understand the role of bats as reservoirs of Bartonella spp., we estimated Bartonella spp. prevalence and genetic diversity in bats in Guatemala during 2009. We found prevalence of 33% and identified 21 genetic variants of 13 phylogroups. Vampire bat-associated Bartonella spp. may cause undiagnosed illnesses in humans. PMID:21762584

  2. Bartonella spp. in Bats, Guatemala

    PubMed Central

    Kosoy, Michael; Recuenco, Sergio; Alvarez, Danilo; Moran, David; Turmelle, Amy; Ellison, James; Garcia, Daniel L.; Estevez, Alejandra; Lindblade, Kim; Rupprecht, Charles

    2011-01-01

    To better understand the role of bats as reservoirs of Bartonella spp., we estimated Bartonella spp. prevalence and genetic diversity in bats in Guatemala during 2009. We found prevalence of 33% and identified 21 genetic variants of 13 phylogroups. Vampire bat–associated Bartonella spp. may cause undiagnosed illnesses in humans. PMID:21762584

  3. Adhesins of Bartonella spp.

    PubMed

    O'Rourke, Fiona; Schmidgen, Thomas; Kaiser, Patrick O; Linke, Dirk; Kempf, Volkhard A J

    2011-01-01

    Adhesion to host cells represents the first step in the infection process and one of the decisive features in the pathogenicity of Bartonella spp. B. henselae and B. quintana are considered to be the most important human pathogenic species, responsible for cat scratch disease, bacillary angiomatosis, trench fever and other diseases. The ability to cause vasculoproliferative disorders and intraerythrocytic bacteraemia are unique features of the genus Bartonella. Consequently, the interaction with endothelial cells and erythrocytes is a focus in Bartonella research. The genus harbours a variety of trimeric autotransporter adhesins (TAAs) such as the Bartonella adhesin A (BadA) of B. henselae and the variably expressed outer-membrane proteins (Vomps) of B. quintana, which display remarkable variations in length and modular construction. These adhesins mediate many of the biologically-important properties of Bartonella spp. such as adherence to endothelial cells and extracellular matrix proteins and induction of angiogenic gene programming. There is also significant evidence that the laterally acquired Trw-conjugation systems of Bartonella spp. mediate host-specific adherence to erythrocytes. Other potential adhesins are the filamentous haemagglutinins and several outer membrane proteins. The exact molecular functions of these adhesins and their interplay with other pathogenicity factors (e.g., the VirB/D4 type 4 secretion system) need to be analysed in detail to understand how these pathogens adapt to their mammalian hosts.

  4. IDENTIFICATION OF DIFFERENT FUSARIUM SPP. IN ALLIUM SPP. IN GERMANY.

    PubMed

    Boehnke, B; Karlovsky, P; Pfohl, K; Gamliel, A; Isack, Y; Dehne, H W

    2015-01-01

    In 2013 Allium cepa bulbs from different fields in Northern and Southern Germany, seeds and sets from onion breeders were analysed for infestation with Fusarium species. The same investigation was done in 2014 with different edible Allium spp. from local markets. Different Fusarium spp. were isolated and identified by morphological characterisation. 24 different Fusarium spp. were identified. The diversity of Fusarium spp. and the intensity of infestation was higher on edible bulbs compared to the younger sets and seeds. The analysed onions and other edible Allium spp. from local markets showed also high contents of different Fusarium species. The most prevalent identified Fusarium sp. in the analysed Allium spp. in Germany was Fusarium oxysporum which can cause the Fusarium Basal Rot, followed by Fusarium solani. Fusarium proliferatum, which can cause the Fusarium Salmon Blotch in onions, could be detected in about half of the sampled onion fields and in approximately 10% of all analysed onions from fields. Also in the onion sets, on the surface of the seeds and in other edible Allium spp. F. proliferatum could be identified. Besides F. proliferatum, further mycotoxin producing Fusarium spp. like Fusarium equiseti or Fusarium tricinctum were identified. Other Fusarium spp. like Fusarium sporotrichioides and Fusarium poae were first described in Allium sp. in this study. The two most prevalent Fusarium spp. F. oxysporum and F. solani are able to produce mycotoxins like enniatins, fumonisins, moniliformin and T-2 toxins. Fusarium sp. like F. proliferatum, F. equiseti and F. tricinctum are able to produce additional toxins like beauvericins, zearalenone and diacetoscirpenol. This high number of Fusarium spp., which are able to produce a broad spectrum of different mycotoxins, could be a potential health risk for human beings and livestock.

  5. Sugarcane (Saccharum spp.).

    PubMed

    Arencibia, Ariel D; Carmona, Elva R

    2006-01-01

    We describe the procedures for recovering transgenic sugarcane from co-cultivation of both calli and in vitro plants with Agrobacterium tumefaciens. The correct tissue culture strategies and the use of super-binary vector or super-virulent strain are crucial for the successful sugarcane transformation. Both plant regeneration via calli culture and micropropagation strategies can be optimized to a wide spectrum of sugarcane genotypes, thus the procedures presented here could be applied to genetic engineering of Saccharum spp. after minor modifications. For the case of sugarcane transformation using in vitro plants, four selective micropropagation steps must be sufficient to eliminate chimera plants.

  6. First isolation of Mycobacterium spp. in Mullus spp. in Turkey

    PubMed Central

    Sevim, P; Ozer, S; Rad, F

    2015-01-01

    Ichthyozoonotic Mycobacterium spp. poses health risks both to fish and humans. In this study, the presence of ichthyozoonotic Mycobacterium spp. was investigated in red mullet (Mullus barbatus barbatus) and surmullet (Mullus surmuletus), widely caught species in the Mediterranean and the Aegean Sea. A total of 208 fish samples, provided from fishermen of Mersin province (Turkey) were studied. Using conventional methods, Mycobacterium spp. was isolated and identified at the genus level by PCR and at the species level by PCR-RFLP. Thirteen Mycobacterium spp. were detected in 13 (6.25%) fish samples. Four mycobacteria were identified as M. genavense, three as M. fortuitum, three as M. scrofulaceum, one as M. marinum, one as M. vaccae and one as M. aurum. No signs of mycobacteriosis were observed in fish samples. Findings of this study can contribute to future studies of onichthyozoonotic Mycobacterium spp. in seafood. PMID:27175166

  7. SPP1-mediated plasmid transduction.

    PubMed Central

    Canosi, U; Lüder, G; Trautner, T A

    1982-01-01

    The virulent Bacillus subtilis phage SPP1 transduces plasmid DNA. Plasmid-transducing phages contain only plasmid DNA. Such DNA represents a concatemer of monomeric plasmid molecules with the molecular weight of mature SPP1 DNA. Biological parameters of plasmid transduction are described. Images PMID:6292508

  8. Humoral antibody response of the tilapia Oreochromis niloticus against Cichlidogyrus spp. (Monogenea).

    PubMed

    Sandoval-Gío, Juan José; Rodríguez-Canul, Rossanna; Vidal-Martínez, Víctor Manuel

    2008-04-01

    The humoral immune response of the tilapia Oreochromis niloticus was evaluated using a direct ELISA. Serum was tested from fish infected with Cichlidogyrus spp. (Monogenea) and from fish injected intraperitoneally with the Cichlidogyrus spp. antigenic extract, i.e., 150 microl of the Cichlidogyrus spp. saline extract diluted in Freund's complete adjuvant (FCA) (1:1) were inoculated intraperitoneally at day 0, followed by 2 dosages of 50 microl of the same Cichlidogyrus spp. saline extract diluted in Freund's incomplete adjuvant (FIA) (1:1) at weeks 2 and 4, respectively. The humoral response was also evaluated by the double immunodiffusion test (DID) and by serum protein and total immunoglobulin (Ig) determinations. The IgM OD values in the hyperimmune fish were significantly higher than in the infected and uninfected fish groups. In the DID test, a precipitation (antigen-antibody) band was observed between the Cichlidogyrus spp. saline extract and hyperimmune sera, but not with the other groups. Increases in serum protein concentration and total Igs were observed in the immunized fish at weeks 2 and 10 postinjection. Results from this study suggest that tilapia is capable of producing an induced humoral immune response against an antigenic extract of Cichlidogyrus spp. PMID:18564741

  9. Detection of Helicobacter spp. DNA in the oral cavity of dogs.

    PubMed

    Recordati, Camilla; Gualdi, Valentina; Tosi, Sabrina; Facchini, Roberto Vailati; Pengo, Graziano; Luini, Mario; Simpson, Kenneth W; Scanziani, Eugenio

    2007-01-31

    The mode of acquisition of gastric Helicobacter spp. infection in dogs has not been determined. It is suspected that oral-oral and faecal-oral transmission may be involved. The present study sought to determine if Helicobacter spp. DNA is present in the oral cavity of healthy and vomiting dogs. Thirty-eight pet dogs (27 vomiting and 11 clinically healthy) were studied. The presence of Helicobacter spp. was determined by single and nested PCR evaluation of DNA extracted from saliva, dental plaque and gastric biopsy samples. Helicobacter spp. DNA was detected by nested PCR in 36 (94.7%) gastric biopsies, 17 (44.7%) dental plaque and 19 (50%) saliva samples out of the 38 dogs examined. Overall 27 (71.1%) dogs screened by nested PCR were found to harbour Helicobacter spp. DNA in the oral cavity (dental plaque and/or saliva). There was no significant difference in the prevalence of Helicobacter spp. DNA in the oral cavity of vomiting and healthy dogs, and the time from vomiting to oral sampling did not have significant impact. This study confirms the high prevalence of gastric Helicobacter spp. infection in dogs, and reveals that Helicobacter spp. DNA is detectable in the oral cavity of over 70% of dogs. These findings support the possibility of oral-oral transmission between dogs and that the canine oral cavity may act as source of non-pylori Helicobacter spp. infection for humans.

  10. Volatiles from a rare Acer spp. honey sample from Croatia.

    PubMed

    Jerković, Igor; Marijanović, Zvonimir; Malenica-Staver, Mladenka; Lusić, Drazen

    2010-06-24

    A rare sample of maple (Acer spp.) honey from Croatia was analysed. Ultrasonic solvent extraction (USE) using: 1) pentane, 2) diethyl ether, 3) a mixture of pentane and diethyl ether (1:2 v/v) and 4) dichloromethane as solvents was applied. All the extracts were analysed by GC and GC/MS. The most representative extracts were 3) and 4). Syringaldehyde was the most striking compound, being dominant in the extracts 2), 3) and 4) with percentages 34.5%, 33.1% and 35.9%, respectively. In comparison to USE results of other single Croatian tree honey samples (Robinia pseudoacacia L. nectar honey, Salix spp. nectar and honeydew honeys, Quercus frainetto Ten. honeydew as well as Abies alba Mill. and Picea abies L. honeydew) and literature data the presence of syringaldehyde, previously identified in maple sap and syrup, can be pointed out as a distinct characteristic of the Acer spp. honey sample. Headspace solid-phase microextraction (HS-SPME) combined with GC and GC/MS identified benzaldehyde (16.5%), trans-linalool oxide (20.5%) and 2-phenylethanol (14.9%) as the major compounds that are common in different honey headspace compositions.

  11. Comparative genomics of Fructobacillus spp. and Leuconostoc spp. reveals niche-specific evolution of Fructobacillus spp.

    DOE PAGES

    Endo, Akihito; Tanizawa, Yasuhiro; Tanaka, Naoto; Maeno, Shintaro; Kumar, Himanshu; Shiwa, Yuh; Okada, Sanae; Yoshikawa, Hirofumi; Dicks, Leon; Nakagawa, Junichi; et al

    2015-12-29

    In this study, Fructobacillus spp. in fructose-rich niches belong to the family Leuconostocaceae. They were originally classified as Leuconostoc spp., but were later grouped into a novel genus, Fructobacillus , based on their phylogenetic position, morphology and specific biochemical characteristics. The unique characters, so called fructophilic characteristics, had not been reported in the group of lactic acid bacteria, suggesting unique evolution at the genome level. Here we studied four draft genome sequences of Fructobacillus spp. and compared their metabolic properties against those of Leuconostoc spp. As a result, Fructobacillus species possess significantly less protein coding sequences in their small genomes.more » The number of genes was significantly smaller in carbohydrate transport and metabolism. Several other metabolic pathways, including TCA cycle, ubiquinone and other terpenoid-quinone biosynthesis and phosphotransferase systems, were characterized as discriminative pathways between the two genera. The adhE gene for bifunctional acetaldehyde/alcohol dehydrogenase, and genes for subunits of the pyruvate dehydrogenase complex were absent in Fructobacillus spp. The two genera also show different levels of GC contents, which are mainly due to the different GC contents at the third codon position. In conclusion, the present genome characteristics in Fructobacillus spp. suggest reductive evolution that took place to adapt to specific niches.« less

  12. Parasitic polymorphism of Coccidioides spp

    PubMed Central

    2014-01-01

    Background Coccidioides spp. is the ethiological agent of coccidioidomycosis, an infection that can be fatal. Its diagnosis is complicated, due to that it shares clinical and histopathological characteristics with other pulmonary mycoses. Coccidioides spp. is a dimorphic fungus and, in its saprobic phase, grows as a mycelium, forming a large amount of arthroconidia. In susceptible persons, arthroconidia induce dimorphic changes into spherules/endospores, a typical parasitic form of Coccidioides spp. In addition, the diversity of mycelial parasitic forms has been observed in clinical specimens; they are scarcely known and produce errors in diagnosis. Methods We presented a retrospective study of images from specimens of smears with 15% potassium hydroxide, cytology, and tissue biopsies of a histopathologic collection from patients with coccidioidomycosis seen at a tertiary-care hospital in Mexico City. Results The parasitic polymorphism of Coccidioides spp. observed in the clinical specimens was as follows: i) spherules/endospores in different maturation stages; ii) pleomorphic cells (septate hyphae, hyphae composed of ovoid and spherical cells, and arthroconidia), and iii) fungal ball formation (mycelia with septate hyphae and arthroconidia). Conclusions The parasitic polymorphism of Coccidioides spp. includes the following: spherules/endospores, arthroconidia, and different forms of mycelia. This knowledge is important for the accurate diagnosis of coccidioidomycosis. In earlier studies, we proposed the integration of this diversity of forms in the Coccidioides spp. parasitic cycle. The microhabitat surrounding the fungus into the host would favor the parasitic polymorphism of this fungus, and this environment may assist in the evolution toward parasitism of Coccidioides spp. PMID:24750998

  13. Infections Caused by Scedosporium spp.

    PubMed Central

    Cortez, Karoll J.; Roilides, Emmanuel; Quiroz-Telles, Flavio; Meletiadis, Joseph; Antachopoulos, Charalampos; Knudsen, Tena; Buchanan, Wendy; Milanovich, Jeffrey; Sutton, Deanna A.; Fothergill, Annette; Rinaldi, Michael G.; Shea, Yvonne R.; Zaoutis, Theoklis; Kottilil, Shyam; Walsh, Thomas J.

    2008-01-01

    Scedosporium spp. are increasingly recognized as causes of resistant life-threatening infections in immunocompromised patients. Scedosporium spp. also cause a wide spectrum of conditions, including mycetoma, saprobic involvement and colonization of the airways, sinopulmonary infections, extrapulmonary localized infections, and disseminated infections. Invasive scedosporium infections are also associated with central nervous infection following near-drowning accidents. The most common sites of infection are the lungs, sinuses, bones, joints, eyes, and brain. Scedosporium apiospermum and Scedosporium prolificans are the two principal medically important species of this genus. Pseudallescheria boydii, the teleomorph of S. apiospermum, is recognized by the presence of cleistothecia. Recent advances in molecular taxonomy have advanced the understanding of the genus Scedosporium and have demonstrated a wider range of species than heretofore recognized. Studies of the pathogenesis of and immune response to Scedosporium spp. underscore the importance of innate host defenses in protection against these organisms. Microbiological diagnosis of Scedosporium spp. currently depends upon culture and morphological characterization. Molecular tools for clinical microbiological detection of Scedosporium spp. are currently investigational. Infections caused by S. apiospermum and P. boydii in patients and animals may respond to antifungal triazoles. By comparison, infections caused by S. prolificans seldom respond to medical therapy alone. Surgery and reversal of immunosuppression may be the only effective therapeutic options for infections caused by S. prolificans. PMID:18202441

  14. Rickettsial pathogens in the tropical rat mite Ornithonyssus bacoti (Acari: Macronyssidae) from Egyptian rats (Rattus spp.).

    PubMed

    Reeves, Will K; Loftis, Amanda D; Szumlas, Daniel E; Abbassy, Magda M; Helmy, Ibrahim M; Hanafi, Hanafi A; Dasch, Gregory A

    2007-01-01

    We collected and tested 616 tropical rat mites (Ornithonyssus bacoti (Hirst)) from rats (Rattus norvegicus (Berkenhout) and R. rattus (Linnaeus)) throughout 14 governorates in Egypt and tested DNA extracts from pools of these mites for Bartonella spp., Coxiella burnetii, and Rickettsia spp. by PCR amplification and sequencing. Three different mite-associated bacterial agents, including one Bartonella and two Rickettsia spp., were detected in eight pools of mites. Further research could demonstrate the vector potential of mites and pathogenicity of these agents to humans or animals. PMID:17225079

  15. A Ribeiroia spp. (Class: Trematoda) - Specific PCR-based diagnostic

    USGS Publications Warehouse

    Reinitz, D.M.; Yoshino, T.P.; Cole, R.A.

    2007-01-01

    Increased reporting of amphibian malformations in North America has been noted with concern in light of reports that amphibian numbers and species are declining worldwide. Ribeiroia ondatrae has been shown to cause a variety of types of malformations in amphibians. However, little is known about the prevalence of R. ondatrae in North America. To aid in conducting field studies of Ribeiroia spp., we have developed a polymerase chain reaction (PCR)-based diagnostic. Herein, we describe the development of an accurate, rapid, simple, and cost-effective diagnostic for detection of Ribeiroia spp. infection in snails (Planorbella trivolvis). Candidate oligonucleotide primers for PCR were designed via DNA sequence analyses of multiple ribosomal internal transcribed spacer-2 regions from Ribeiroia spp. and Echinostoma spp. Comparison of consensus sequences determined from both genera identified areas of sequence potentially unique to Ribeiroia spp. The PCR reliably produced a diagnostic 290-base pair (bp) product in the presence of a wide concentration range of snail or frog DNA. Sensitivity was examined with DNA extracted from single R. ondatrae cercaria. The single-tube PCR could routinely detect less than 1 cercariae equivalent, because DNA isolated from a single cercaria could be diluted at least 1:50 and still yield a positive result via gel electrophoresis. An even more sensitive nested PCR also was developed that routinely detected 100 fg of the 290-bp fragment. The assay did not detect furcocercous cercariae of certain Schistosomatidae, Echinostoma sp., or Sphaeridiotrema globulus nor adults of Clinostomum sp. or Cyathocotyle bushiensis. Field testing of 137 P. trivolvis identified 3 positives with no overt environmental cross-reactivity, and results concurred with microscopic examinations in all cases. ?? American Society of Parasitologists 2007.

  16. Diseases of tunas, Thunnus spp.

    PubMed

    Munday, B L; Sawada, Y; Cribb, T; Hayward, C J

    2003-04-01

    Much is known about those aspects of tuna health which can be studied in wild populations, e.g. helminth parasites. However, because aquaculture of these species is in its infancy, knowledge of microbial, nutritional and environmental diseases is limited. This review is an attempt to bring together the available information on those diseases of Thunnus spp. which cause significant morbidity, mortality or economic loss. In doing so it has become clear that much more research needs to be undertaken on the physiology of the species (southern, northern and Pacific bluefin tuna) currently used in aquaculture in order for the pathogenesis of some conditions to be properly understood. Attempts at hatchery culture of Pacific bluefin tuna has indicated that Thunnus spp. will be problematic to hatch and propagate.

  17. Genetic engineering of Geobacillus spp.

    PubMed

    Kananavičiūtė, Rūta; Čitavičius, Donaldas

    2015-04-01

    Members of the genus Geobacillus are thermophiles that are of great biotechnological importance, since they are sources of many thermostable enzymes. Because of their metabolic versatility, geobacilli can be used as whole-cell catalysts in processes such as bioconversion and bioremediation. The effective employment of Geobacillus spp. requires the development of reliable methods for genetic engineering of these bacteria. Currently, genetic manipulation tools and protocols are under rapid development. However, there are several convenient cloning vectors, some of which replicate autonomously, while others are suitable for the genetic modification of chromosomal genes. Gene expression systems are also intensively studied. Combining these tools together with proper techniques for DNA transfer, some Geobacillus strains were shown to be valuable producers of recombinant proteins and industrially important biochemicals, such as ethanol or isobutanol. This review encompasses the progress made in the genetic engineering of Geobacillus spp. and surveys the vectors and transformation methods that are available for this genus.

  18. The biology of Giardia spp.

    PubMed Central

    Adam, R D

    1991-01-01

    Gardia spp. are flagellated protozoans that parasitize the small intestines of mammals, birds, reptiles, and amphibians. The infectious cysts begin excysting in the acidic environment of the stomach and become trophozoites (the vegetative form). The trophozoites attach to the intestinal mucosa through the suction generated by a ventral disk and cause diarrhea and malabsorption by mechanisms that are not well understood. Giardia spp. have a number of unique features, including a predominantly anaerobic metabolism, complete dependence on salvage of exogenous nucleotides, a limited ability to synthesize and degrade carbohydrates and lipids, and two nuclei that are equal by all criteria that have been tested. The small size and unique sequence of G. lamblia rRNA molecules have led to the proposal that Giardia is the most primitive eukaryotic organism. Three Giardia spp. have been identified by light lamblia, G. muris, and G. agilis, but electron microscopy has allowed further species to be described within the G. lamblia group, some of which have been substantiated by differences in the rDNA. Animal models and human infections have led to the conclusion that intestinal infection is controlled primarily through the humoral immune system (T-cell dependent in the mouse model). A major immunogenic cysteine-rich surface antigen is able to vary in vitro and in vivo in the course of an infection and may provide a means of evading the host immune response or perhaps a means of adapting to different intestinal environments. Images PMID:1779932

  19. Survey of Legionella spp. in Mud Spring Recreation Area

    NASA Astrophysics Data System (ADS)

    Hsu, B.-M.; Ma, P.-H.; Su, I.-Z.; Chen, N.-S.

    2009-04-01

    Legionella genera are parasites of FLA, and intracellular bacterial replication within the FLA plays a major role in the transmission of disease. At least 13 FLA species—including Acanthamoeba spp., Naegleria spp., and Hartmannella spp.—support intracellular bacterial replication. In the study, Legionellae were detected with microbial culture or by direct DNA extraction and analysis from concentrated water samples or cultured free-living amoebae, combined with molecular methods that allow the taxonomic identification of these pathogens. The water samples were taken from a mud spring recreation area located in a mud-rock-formation area in southern Taiwan. Legionella were detected in 15 of the 34 samples (44.1%). Four of the 34 samples analyzed by Legionella culture were positive for Legionella, five of 34 were positive for Legionella when analyzed by direct DNA extraction and analysis, and 11 of 34 were positive for amoebae-resistant Legionella when analyzed by FLA culture. Ten samples were shown to be positive for Legionella by one analysis method and five samples were shown to be positive by two analysis methods. However, Legionella was detected in no sample by all three analysis methods. This suggests that the three analysis methods should be used together to detect Legionella in aquatic environments. In this study, L. pneumophila serotype 6 coexisted with A. polyphaga, and two uncultured Legionella spp. coexisted with either H. vermiformis or N. australiensis. Of the unnamed Legionella genotypes detected in six FLA culture samples, three were closely related to L. waltersii and the other three were closely related to L. pneumophila serotype 6. Legionella pneumophila serotype 6, L. drancourtii, and L. waltersii are noted endosymbionts of FLA and are categorized as pathogenic bacteria. This is significant for human health because these Legionella exist within FLA and thus come into contact with typically immunocompromised people.

  20. [Mycoses and zoonoses: Cryptococcus spp].

    PubMed

    Cabañes, F Javier

    2008-03-01

    The term "zoonosis" is difficult to delimit because different authors have various definitions for this term. Few mycoses are usually considered zoonoses. However, the role that animals play in the epidemiology of the main human mycoses is still not well known. Moreover, the environmental niches for these fungal agents have not yet been completely determined. This special issue of the "Revista Iberoamericana de Micología" deals with the talks and round table presented at the VIII Spanish Mycological Congress held in October 2006 in Barcelona, Spain on "Cryptococcus spp. and zoonoses".

  1. Osteosarcoma in Baboons (Papio spp)

    PubMed Central

    Mezzles, Marguerite J; Dick, Edward J; Owston, Michael A; Bauer, Cassondra

    2015-01-01

    Bone neoplasms in baboons (Papio spp) are rare, with only one confirmed case of osteosarcoma previously described in the literature. Over a 12-y period, 6 baboons at a national primate research center presented with naturally occurring osteosarcoma; 3 lesions affected the appendicular skeleton, and the remaining 3 were in the head (skull and mandible). The 6 cases presented were identified in members of a large outdoor-housed breeding colony. The subjects were not genetically related or exposed to the same research conditions. Diagnoses were made based on the presentation and radiographic findings, with histologic confirmation. PMID:25926401

  2. Osteosarcoma in Baboons (Papio spp).

    PubMed

    Mezzles, Marguerite J; Dick, Edward J; Owston, Michael A; Bauer, Cassondra

    2015-04-01

    Bone neoplasms in baboons (Papio spp) are rare, with only one confirmed case of osteosarcoma previously described in the literature. Over a 12-y period, 6 baboons at a national primate research center presented with naturally occurring osteosarcoma; 3 lesions affected the appendicular skeleton, and the remaining 3 were in the head (skull and mandible). The 6 cases presented were identified in members of a large outdoor-housed breeding colony. The subjects were not genetically related or exposed to the same research conditions. Diagnoses were made based on the presentation and radiographic findings, with histologic confirmation.

  3. Synanthropic Cockroaches (Blattidae: Periplaneta spp.) Harbor Pathogenic Leptospira in Colombia.

    PubMed

    Gonzalez-Astudillo, Viviana; Bustamante-Rengifo, Javier A; Bonilla, Álvaro; Lehmicke, Anna Joy J; Castillo, Andrés; Astudillo-Hernández, Miryam

    2016-01-01

    Leptospirosis cases in Colombia are typically linked to peridomestic rodents; however, empirical data suggest that Leptospira-infected patients with no apparent exposure to these reservoirs are common. Cockroaches (Periplaneta spp.) have equal or greater interaction with humans than rodents, yet their potential role as carriers of Leptospira has not been assessed. We determined if pathogenic Leptospira is harbored by Periplaneta spp. in Cali (Colombia) and the variables influencing this relationship. Fifty-nine cockroaches were captured from seven sites and DNA was extracted from the body surface and digestive tract for a multiplex polymerase chain reaction, targeting genes secY and flaB. Logistic regression models and proportion tests showed a higher likelihood for Leptospira to be isolated from body surfaces (P > 0.001) and from individuals inside houses (six times more likely). These findings are the first to demonstrate an association between Periplaneta spp. and Leptospira, suggesting the need to investigate the potential for cockroaches to serve as reservoirs or transport hosts for Leptospira.

  4. The novel Shewanella putrefaciens-infecting bacteriophage Spp001: genome sequence and lytic enzymes.

    PubMed

    Han, Feng; Li, Meng; Lin, Hong; Wang, Jingxue; Cao, Limin; Khan, Muhammad Naseem

    2014-06-01

    Shewanella putrefaciens has been identified as a specific spoilage organism commonly found in chilled fresh fish, which contributes to the spoilage of fish products. Limiting S. putrefaciens growth can extend the shelf-life of chilled fish. Endolysins, which are lytic enzymes produced by bacteriophages, have been considered an alternative to control bacterial growth, and have been useful in various applications, including food preservation. We report here, for the first time, the complete genome sequence of a novel phage Spp001, which lyses S. putrefaciens Sp225. The Spp001 genome comprises a 54,789-bp DNA molecule with 67 open reading frames and an average total G + C content of 49.42 %. In silico analysis revealed that the Spp001 open reading frames encode various putative functional proteins, including an endolysin (ORF 62); however, no sequence for genes encoding the holin polypeptides, which work in concert with endolysins, was identified. To examine further the lytic activity of Spp001, we analyzed the lytic enzyme-containing fraction from phages released at the end of the phage lytic cycle in S. putrefaciens, using diffusion and turbidimetric assays. The results show that the partially purified extract contained endolysin, as indicated by a high hydrolytic activity towards bacterial peptidoglycan decrease in the OD590 value by 0.160 in 15 min. The results will allow further investigation of the purification of natural Spp001 endolysin, the extension of Spp001 host range, and the applications of the phage-encoded enzymes. PMID:24740748

  5. The novel Shewanella putrefaciens-infecting bacteriophage Spp001: genome sequence and lytic enzymes.

    PubMed

    Han, Feng; Li, Meng; Lin, Hong; Wang, Jingxue; Cao, Limin; Khan, Muhammad Naseem

    2014-06-01

    Shewanella putrefaciens has been identified as a specific spoilage organism commonly found in chilled fresh fish, which contributes to the spoilage of fish products. Limiting S. putrefaciens growth can extend the shelf-life of chilled fish. Endolysins, which are lytic enzymes produced by bacteriophages, have been considered an alternative to control bacterial growth, and have been useful in various applications, including food preservation. We report here, for the first time, the complete genome sequence of a novel phage Spp001, which lyses S. putrefaciens Sp225. The Spp001 genome comprises a 54,789-bp DNA molecule with 67 open reading frames and an average total G + C content of 49.42 %. In silico analysis revealed that the Spp001 open reading frames encode various putative functional proteins, including an endolysin (ORF 62); however, no sequence for genes encoding the holin polypeptides, which work in concert with endolysins, was identified. To examine further the lytic activity of Spp001, we analyzed the lytic enzyme-containing fraction from phages released at the end of the phage lytic cycle in S. putrefaciens, using diffusion and turbidimetric assays. The results show that the partially purified extract contained endolysin, as indicated by a high hydrolytic activity towards bacterial peptidoglycan decrease in the OD590 value by 0.160 in 15 min. The results will allow further investigation of the purification of natural Spp001 endolysin, the extension of Spp001 host range, and the applications of the phage-encoded enzymes.

  6. Prevalence of Microsporidia, Cryptosporidium spp., and Giardia spp. in beavers (Castor canadensis) in Massachusetts

    USGS Publications Warehouse

    Fayer, R.; Santin, M.; Trout, J.M.; DeStefano, S.; Koenen, K.; Kaur, T.

    2006-01-01

    Feces from 62 beavers (Castor canadensis) in Massachusetts were examined by fluorescence microscopy (IFA) and polymerase chain reaction (PCR) for Microsporidia species, Cryptosporidium spp., and Giardia spp. between January 2002 and December 2004. PCR-positive specimens were further examined by gene sequencing. Protist parasites were detected in 6.4% of the beavers. All were subadults and kits. Microsporidia species were not detected. Giardia spp. was detected by IFA from four beavers; Cryptosporidium spp. was also detected by IFA from two of these beavers. However, gene sequence data for the ssrRNA gene from these two Cryptosporidium spp.-positive beavers were inconclusive in identifying the species. Nucleotide sequences of the TPI, ssrRNA, and ??-giardin genes for Giardia spp. (deposited in GenBank) indicated that the four beavers were excreting Giardia duodenalis Assemblage B, the zoonotic genotype representing a potential source of waterborne Giardia spp. cysts. Copyright 2006 by American Association of Zoo Veterinarians.

  7. Haloalkaliphilic Streptomyces spp. AJ8 isolated from solar salt works and its' pharmacological potential.

    PubMed

    Jenifer, John Selesteen Charles Adlin; Donio, Mariathason Birdilla Selva; Michaelbabu, Mariavincent; Vincent, Samuel Gnana Prakash; Citarasu, Thavasimuthu

    2015-12-01

    Antagonistic Streptomyces spp. AJ8 was isolated and identified from the Kovalam solar salt works in India. The antimicrobial NRPS cluster gene was characterized by PCR, sequencing and predict the secondary structure analysis. The secondary metabolites will be extracted from different organic solvent extraction and studied the antibacterial, antifungal, antiviral and anticancer activities. In vitro antagonistic activity results revealed that, Streptomyces spp. AJ8 was highly antagonistic against Staphylococcus aureus, Aeromonas hydrophila WPD1 and Candida albicans. The genomic level identification revealed that, the strain was confirmed as Streptomyces spp. AJ8 and submitted the NCBI database (KC603899). The NRPS gene was generated a single gene fragment of 781 bp length (KR491940) and the database analysis revealed that, the closely related to Streptomyces spp. SAUK6068 and S. coeruleoprunus NBRC15400. The secondary metabolites extracted with ethyl acetate was effectively inhibited the bacterial and fungal growth at the ranged between 7 and 19.2 mm of zone of inhibition. The antiviral activity results revealed that, the metabolite was significantly (P < 0.001) controlled the killer shrimp virus white spot syndrome virus at the level of 85 %. The metabolite also suppressed the L929 fibroblast cancer cells at 35.7 % viability in 1000 µg treatment.

  8. Presence of Salmonella spp. and Campylobacter spp. in shellfish.

    PubMed

    Wilson, I G; Moore, J E

    1996-04-01

    Bivalve molluscs, (cockles, mussels, scallops and oysters) were examined according to EC shellfish bed classification regulations for faecal coliforms, Escherichia coli and salmonella, and for coliforms and campylobacter which are not specified by these regulations. Salmonella serotypes were detected in 8% of 433 molluscs. Seven salmonella isolations (2%) were made from category A beds, nominally suitable for immediate consumption according to E. coli counts. A higher percentage of salmonella isolates (6%) was detected in shellfish which require relaying or depuration prior to eating. In another survey, thermophilic Campylobacter spp. were found in 42% of 380 shellfish. These findings show bed classification on the basis of indicator organisms alone is not sufficient to assure the absence of bacterial, and no doubt viral, pathogens. Depuration and end product specifications which require the absence of salmonellae are an essential part of these regulations. Microbiologists may wish to consider whether tests for pathogens such as salmonella and campylobacter should be included when determining the suitability of shellfish for human consumption. PMID:8620905

  9. Absence of serological evidence of Rickettsia spp., Bartonella spp., Ehrlichia spp. and Coxiella burnetii infections in American Samoa.

    PubMed

    Lau, Colleen; Musso, Didier; Fournier, Pierre-Edouard; Parola, Philippe; Raoult, Didier; Weinstein, Philip

    2016-07-01

    Little is known about the epidemiology of zoonotic diseases in American Samoa (Pacific). A review of literature did not identify any published information on human Rickettsia spp., Bartonella spp., Ehrlichia spp. or Coxiella burnetii infections in this country. To determine the presence of these diseases, we conducted a serosurvey of American Samoans. The presence of immunoglobulin G antibodies against Rickettsia felis, Rickettsia typhi, Rickettsia conorii, C. burnetii, Bartonella henselae, Bartonella quintana, and Ehrlichia chaffeensis was evaluated by indirect immunofluorescence assay in sera from 197 American Samoan adults. None of the samples had antibodies at a significant level against Rickettsia spp., Bartonella spp., Ehrlichia spp. or C. burnetii (seroprevalence 0%; one-tailed 95% CI 0-1.86%). We cannot conclude that these pathogens are absent in American Samoa but, if present, their prevalence is probably very low. Q fever has been reported worldwide except in New Zealand and French Polynesia; these new data suggest that the prevalence of Q fever is likely to be very low in the Pacific Islands. PMID:26965788

  10. Mycotoxins produced by Fusarium spp. associated with Fusarium head blight of wheat in Western Australia.

    PubMed

    Tan, Diana C; Flematti, Gavin R; Ghisalberti, Emilio L; Sivasithamparam, Krishnapillai; Chakraborty, Sukumar; Obanor, Friday; Jayasena, Kithsiri; Barbetti, Martin J

    2012-05-01

    An isolated occurrence of Fusarium head blight (FHB) of wheat was detected in the south-west region of Western Australia during the 2003 harvest season. The molecular identity of 23 isolates of Fusarium spp. collected from this region during the FHB outbreak confirmed the associated pathogens to be F. graminearum, F. acuminatum or F. tricinctum. Moreover, the toxicity of their crude extracts from Czapek-Dox liquid broth and millet seed cultures to brine shrimp (Artemia franciscana) was associated with high mortality levels. The main mycotoxins detected were type B trichothecenes (deoxynivalenol and 3-acetyldeoxynivalenol), enniatins, chlamydosporol and zearalenone. This study is the first report on the mycotoxin profiles of Fusarium spp. associated with FHB of wheat in Western Australia. This study highlights the need for monitoring not just for the presence of the specific Fusarium spp. present in any affected grain but also for their potential mycotoxin and other toxic secondary metabolites. PMID:23606046

  11. Bartonella spp. DNA associated with biting flies from California.

    PubMed

    Chung, Crystal Y; Kasten, Rickie W; Paff, Sandra M; Van Horn, Brian A; Vayssier-Taussat, Muriel; Boulouis, Henri-Jean; Chomel, Bruno B

    2004-07-01

    Bartonella DNA was investigated in 104 horn flies (Haematobia spp.), 60 stable flies (Stomoxys spp.), 11 deer flies (Chrysops spp.), and 11 horse flies (Tabanus spp.) collected on cattle in California. Partial sequencing indicated B. bovis DNA in the horn fly pool and B. henselae type M DNA in one stable fly. PMID:15324557

  12. WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL.

    PubMed

    Tiyo, Rogerio; de Souza, Carla Zangari; Nishi, Letícia; Brustolin, Camila Fernanda; Ratti, Bianca Altrão; Falavigna Guilherme, Ana Lucia

    2015-01-01

    The aim of this work was to compare, from a parasitological ( Cryptosporidium spp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidium spp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardia spp. or Cryptosporidium spp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidium spp. and Giardia spp., as well as artesian wells for coliform control in local production of vegetables to be marketed. PMID:26422158

  13. WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL.

    PubMed

    Tiyo, Rogerio; de Souza, Carla Zangari; Nishi, Letícia; Brustolin, Camila Fernanda; Ratti, Bianca Altrão; Falavigna Guilherme, Ana Lucia

    2015-01-01

    The aim of this work was to compare, from a parasitological ( Cryptosporidium spp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidium spp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardia spp. or Cryptosporidium spp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidium spp. and Giardia spp., as well as artesian wells for coliform control in local production of vegetables to be marketed.

  14. Longitudinal prevalence, faecal shedding and molecular characterisation of Campylobacter spp. and Salmonella enterica in sheep.

    PubMed

    Yang, Rongchang; Jacobson, Caroline; Gardner, Graham; Carmichael, Ian; Campbell, Angus J D; Ryan, Una

    2014-11-01

    Faecal excretion of Campylobacter spp. and Salmonella enterica in sheep in Australia was determined using a quantitative multiplex PCR (qPCR) targeting the Campylobacter spp. purine biosynthesis gene (PurA) and the S. enterica outer membrane protein (ompF). The mutiplex qPCR was specific and Campylobacter spp. and S. enterica were each detected with a sensitivity of 5 organisms/µL faecal DNA extract. This multiplex qPCR was used to determine the prevalence and concentration of Campylobacter spp. and S. enterica in 3412 faecal samples collected from 1189 lambs on eight farms across South Australia (n = 2 farms), New South Wales (n = 1), Victoria (n = 2) and Western Australia (n = 3) at three sampling periods (weaning, post-weaning and pre-slaughter). The overall prevalences of Campylobacter spp. and S. enterica were 13.3% and 5.0%, respectively, with the highest prevalence for Campylobacter spp. in South Australia and the highest prevalence for S. enterica in New South Wales. Campylobacter jejuni was the only Campylobacter sp. identified from a subset of 120 positive samples sequenced at the 16S locus. S. enterica serovar Typhimurium was the only serovar of S. enterica identified from a subset of 120 positive samples sequenced at the ompF locus. Across all states, Campylobacter spp. had the highest median bacterial concentration in faeces at weaning and post-weaning (medians of 3.4 × 10(6) and 1.1 × 10(5), respectively), whereas S. enterica had the highest median bacterial concentration at pre-slaughter (1.8 × 10(5)/g faeces).

  15. Genetic manipulation of Methanosarcina spp.

    PubMed Central

    Kohler, Petra R. A.; Metcalf, William W.

    2012-01-01

    The discovery of the third domain of life, the Archaea, is one of the most exciting findings of the last century. These remarkable prokaryotes are well known for their adaptations to extreme environments; however, Archaea have also conquered moderate environments. Many of the archaeal biochemical processes, such as methane production, are unique in nature and therefore of great scientific interest. Although formerly restricted to biochemical and physiological studies, sophisticated systems for genetic manipulation have been developed during the last two decades for methanogenic archaea, halophilic archaea and thermophilic, sulfur-metabolizing archaea. The availability of these tools has allowed for more complete studies of archaeal physiology and metabolism and most importantly provides the basis for the investigation of gene expression, regulation and function. In this review we provide an overview of methods for genetic manipulation of Methanosarcina spp., a group of methanogenic archaea that are key players in the global carbon cycle and which can be found in a variety of anaerobic environments. PMID:22837755

  16. Intermolecular interactions of the malate synthase of Paracoccidioides spp

    PubMed Central

    2013-01-01

    Background The fungus Paracoccidioides spp is the agent of paracoccidioidomycosis (PCM), a pulmonary mycosis acquired by the inhalation of fungal propagules. Paracoccidioides malate synthase (PbMLS) is important in the infectious process of Paracoccidioides spp because the transcript is up-regulated during the transition from mycelium to yeast and in yeast cells during phagocytosis by murine macrophages. In addition, PbMLS acts as an adhesin in Paracoccidioides spp. The evidence for the multifunctionality of PbMLS indicates that it could interact with other proteins from the fungus and host. The objective of this study was to identify and analyze proteins that possibly bind to PbMLS (PbMLS-interacting proteins) because protein interactions are intrinsic to cell processes, and it might be possible to infer the function of a protein through the identification of its ligands. Results The search for interactions was performed using an in vivo assay with a two-hybrid library constructed in S. cerevisiae; the transcripts were sequenced and identified. In addition, an in vitro assay using pull-down GST methodology with different protein extracts (yeast, mycelium, yeast-secreted proteins and macrophage) was performed, and the resulting interactions were identified by mass spectrometry (MS). Some of the protein interactions were confirmed by Far-Western blotting using specific antibodies, and the interaction of PbMLS with macrophages was validated by indirect immunofluorescence and confocal microscopy. In silico analysis using molecular modeling, dynamics and docking identified the amino acids that were involved in the interactions between PbMLS and PbMLS-interacting proteins. Finally, the interactions were visualized graphically using Osprey software. Conclusion These observations indicate that PbMLS interacts with proteins that are in different functional categories, such as cellular transport, protein biosynthesis, modification and degradation of proteins and signal

  17. Identification of B-type procyanidins in Fallopia spp. involved in biological denitrification inhibition.

    PubMed

    Bardon, Clément; Piola, Florence; Haichar, Feth el Zahar; Meiffren, Guillaume; Comte, Gilles; Missery, Boris; Balby, Manon; Poly, Franck

    2016-02-01

    Nitrogen (N) is considered as a main limiting factor in plant growth, and nitrogen losses through denitrification can be responsible for severe decreases in plant productivity. Recently, it was demonstrated that Fallopia spp. is responsible for biological denitrification inhibition (BDI) through the release of unknown secondary metabolites. Here, we investigate the secondary metabolites involved in the BDI of Fallopia spp. The antioxidant, protein precipitation capability of Fallopia spp. extracts was measured in relation to the aerobic respiration and denitrification of two bacteria (Gram positive and Gram negative). Proanthocyanidin concentrations were estimated. Proanthocyanidins in extracts were characterized by chromatographic analysis, purified and tested on the bacterial denitrification and aerobic respiration of two bacterial strains. The effect of commercial procyanidins on denitrification was tested on two different soil types. Denitrification and aerobic respiration inhibition were correlated with protein precipitation capacity and concentration of proanthocyanidins but not to antioxidant capacity. These proanthocyanidins were B-type procyanidins that inhibited denitrification more than the aerobic respiration of bacteria. In addition, procyanidins also inhibited soil microbial denitrification. We demonstrate that procyanidins are involved in the BDI of Fallopia spp. Our results pave the way to a better understanding of plant-microbe interactions and highlight future applications for a more sustainable agriculture.

  18. Occurrence of Babesia spp., Rickettsia spp. and Bartonella spp. in Ixodes ricinus in Bavarian public parks, Germany

    PubMed Central

    2011-01-01

    Background Only limited information is available about the occurrence of ticks and tick-borne pathogens in public parks, which are areas strongly influenced by human beings. For this reason, Ixodes ricinus were collected in public parks of different Bavarian cities in a 2-year survey (2009 and 2010) and screened for DNA of Babesia spp., Rickettsia spp. and Bartonella spp. by PCR. Species identification was performed by sequence analysis and alignment with existing sequences in GenBank. Additionally, coinfections with Anaplasma phagocytophilum were investigated. Results The following prevalences were detected: Babesia spp.: 0.4% (n = 17, including one pool of two larvae) in 2009 and 0.5 to 0.7% (n = 11, including one pool of five larvae) in 2010; Rickettsia spp.: 6.4 to 7.7% (n = 285, including 16 pools of 76 larvae) in 2009. DNA of Bartonella spp. in I. ricinus in Bavarian public parks could not be identified. Sequence analysis revealed the following species: Babesia sp. EU1 (n = 25), B. divergens (n = 1), B. divergens/capreoli (n = 1), B. gibsoni-like (n = 1), R. helvetica (n = 272), R. monacensis IrR/Munich (n = 12) and unspecified R. monacensis (n = 1). The majority of coinfections were R. helvetica with A. phagocytophilum (n = 27), but coinfections between Babesia spp. and A. phagocytophilum, or Babesia spp. and R. helvetica were also detected. Conclusions I. ricinus ticks in urban areas of Germany harbor several tick-borne pathogens and coinfections were also observed. Public parks are of particularly great interest regarding the epidemiology of tick-borne pathogens, because of differences in both the prevalence of pathogens in ticks as well as a varying species arrangement when compared to woodland areas. The record of DNA of a Babesia gibsoni-like pathogen detected in I. ricinus suggests that I. ricinus may harbor and transmit more Babesia spp. than previously known. Because of their high recreational value for human beings, urban green areas are likely to

  19. Transferability of SSR and RGA markers developed in Cynodon spp. to Zoysia spp.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bermudagrass (Cynodon spp.) and zoysiagrass (Zoysia spp.), which are both used as warm-season turfgrasses in the United States, are members of subfamily Chloridoideae and are reported to be at least 55% genetically similar. To assess if molecular tools between the two species can be interchanged, 93...

  20. A molecular survey of Babesia spp. and Theileria spp. in red foxes (Vulpes vulpes) and their ticks from Thuringia, Germany.

    PubMed

    Najm, Nour-Addeen; Meyer-Kayser, Elisabeth; Hoffmann, Lothar; Herb, Ingrid; Fensterer, Veronika; Pfister, Kurt; Silaghi, Cornelia

    2014-06-01

    Wild canines which are closely related to dogs constitute a potential reservoir for haemoparasites by both hosting tick species that infest dogs and harbouring tick-transmitted canine haemoparasites. In this study, the prevalence of Babesia spp. and Theileria spp. was investigated in German red foxes (Vulpes vulpes) and their ticks. DNA extracts of 261 spleen samples and 1953 ticks included 4 tick species: Ixodes ricinus (n=870), I. canisuga (n=585), I. hexagonus (n=485), and Dermacentor reticulatus (n=13) were examined for the presence of Babesia/Theileria spp. by a conventional PCR targeting the 18S rRNA gene. One hundred twenty-one out of 261 foxes (46.4%) were PCR-positive. Out of them, 44 samples were sequenced, and all sequences had 100% similarity to Theileria annae. Similarly, sequencing was carried out for 65 out of 118 PCR-positive ticks. Theileria annae DNA was detected in 61.5% of the sequenced samples, Babesia microti DNA was found in 9.2%, and Babesia venatorum in 7.6% of the sequenced samples. The foxes were most positive in June and October, whereas the peak of tick positivity was in October. Furthermore, the positivity of the ticks was higher for I. canisuga in comparison to the other tick species and for nymphs in comparison to adults. The high prevalence of T. annae DNA in red foxes in this study suggests a reservoir function of those animals for T. annae. To our knowledge, this is the first report of T. annae in foxes from Germany as well as the first detection of T. annae and B. microti in the fox tick I. canisuga. Detection of DNA of T. annae and B. microti in three tick species collected from foxes adds new potential vectors for these two pathogens and suggests a potential role of the red fox in their natural endemic cycles.

  1. Minimum inhibitory concentration distribution in environmental Legionella spp. isolates.

    PubMed

    Sandalakis, Vassilios; Chochlakis, Dimosthenis; Goniotakis, Ioannis; Tselentis, Yannis; Psaroulaki, Anna

    2014-12-01

    In Greece standard tests are performed in the watering and cooling systems of hotels' units either as part of the surveillance scheme or following human infection. The purpose of this study was to establish the minimum inhibitory concentration (MIC) distributions of environmental Legionella isolates for six antimicrobials commonly used for the treatment of Legionella infections, by MIC-test methodology. Water samples were collected from 2004 to 2011 from 124 hotels from the four prefectures of Crete (Greece). Sixty-eight (68) Legionella isolates, comprising L. pneumophila serogroups 1, 2, 3, 5, 6, 8, 12, 13, 15, L. anisa, L. rubrilucens, L. maceachernii, L. quinlivanii, L. oakridgensis, and L. taurinensis, were included in the study. MIC-tests were performed on buffered charcoal yeast extract with α-ketoglutarate, L-cysteine, and ferric pyrophosphate. The MICs were read after 2 days of incubation at 36 ± 1 °C at 2.5% CO2. A large distribution in MICs was recorded for each species and each antibiotic tested. Rifampicin proved to be the most potent antibiotic regardless of the Legionella spp.; tetracycline appeared to have the least activity on our environmental isolates. The MIC-test approach is an easy, although not so cost-effective, way to determine MICs in Legionella spp. These data should be kept in mind especially since these Legionella species may cause human disease. PMID:25473976

  2. Minimum inhibitory concentration distribution in environmental Legionella spp. isolates.

    PubMed

    Sandalakis, Vassilios; Chochlakis, Dimosthenis; Goniotakis, Ioannis; Tselentis, Yannis; Psaroulaki, Anna

    2014-12-01

    In Greece standard tests are performed in the watering and cooling systems of hotels' units either as part of the surveillance scheme or following human infection. The purpose of this study was to establish the minimum inhibitory concentration (MIC) distributions of environmental Legionella isolates for six antimicrobials commonly used for the treatment of Legionella infections, by MIC-test methodology. Water samples were collected from 2004 to 2011 from 124 hotels from the four prefectures of Crete (Greece). Sixty-eight (68) Legionella isolates, comprising L. pneumophila serogroups 1, 2, 3, 5, 6, 8, 12, 13, 15, L. anisa, L. rubrilucens, L. maceachernii, L. quinlivanii, L. oakridgensis, and L. taurinensis, were included in the study. MIC-tests were performed on buffered charcoal yeast extract with α-ketoglutarate, L-cysteine, and ferric pyrophosphate. The MICs were read after 2 days of incubation at 36 ± 1 °C at 2.5% CO2. A large distribution in MICs was recorded for each species and each antibiotic tested. Rifampicin proved to be the most potent antibiotic regardless of the Legionella spp.; tetracycline appeared to have the least activity on our environmental isolates. The MIC-test approach is an easy, although not so cost-effective, way to determine MICs in Legionella spp. These data should be kept in mind especially since these Legionella species may cause human disease.

  3. Effective generation of unidirectional SPP beam with arbitrary profile

    NASA Astrophysics Data System (ADS)

    You, Oubo; Bai, Benfeng; Wu, Xiaoyu; Zhu, Zhendong; Wang, Qixia

    2016-04-01

    The beam formation of SPPs is very important in plasmonics. Different SPP beams could be used for different purposes, such as SPP focusing, non-diffractive SPP wave propagation, efficient SPP coupling, and manipulating particles. Here, we present a straightforward and effective method for generating unidirectionally propagating SPP beams with arbitrary profile in both amplitude and phase by locating the Δ-shaped nanoantennas. The Δ-shape of the nanoantennas is used to achieve unidirectionality of SPPs and the locations of the nanoantennas are controlled to realize arbitrary profile of the excited SPP wave. As examples, several SPP launchers generating different SPP beams are designed with this method. The near-field distribution of the generated SPP beams are also experimentally characterized to validate the effectiveness of this method.

  4. Campylobacter spp., Giardia spp., Cryptosporidium spp., noroviruses, and indicator organisms in surface water in southwestern Finland, 2000-2001.

    PubMed

    Hörman, Ari; Rimhanen-Finne, Ruska; Maunula, Leena; von Bonsdorff, Carl-Henrik; Torvela, Niina; Heikinheimo, Annamari; Hänninen, Marja-Liisa

    2004-01-01

    A total of 139 surface water samples from seven lakes and 15 rivers in southwestern Finland were analyzed during five consecutive seasons from autumn 2000 to autumn 2001 for the presence of various enteropathogens (Campylobacter spp., Giardia spp., Cryptosporidium spp., and noroviruses) and fecal indicators (thermotolerant coliforms, Escherichia coli, Clostridium perfringens, and F-RNA bacteriophages) and for physicochemical parameters (turbidity and temperature); this was the first such systematic study. Altogether, 41.0% (57 of 139) of the samples were positive for at least one of the pathogens; 17.3% were positive for Campylobacter spp. (45.8% of the positive samples contained Campylobacter jejuni, 25.0% contained Campylobacter lari, 4.2% contained Campylobacter coli, and 25.0% contained Campylobacter isolates that were not identified), 13.7% were positive for Giardia spp., 10.1% were positive for Cryptosporidium spp., and 9.4% were positive for noroviruses (23.0% of the positive samples contained genogroup I and 77.0% contained genogroup II). The samples were positive for enteropathogens significantly (P < 0.05) less frequently during the winter season than during the other sampling seasons. No significant differences in the prevalence of enteropathogens were found when rivers and lakes were compared. The presence of thermotolerant coliforms, E. coli, and C. perfringens had significant bivariate nonparametric Spearman's rank order correlation coefficients (P < 0.001) with samples that were positive for one or more of the pathogens analyzed. The absence of these indicators in a logistic regression model was found to have significant predictive value (odds ratios, 1.15 x 10(8), 7.57, and 2.74, respectively; P < 0.05) for a sample that was negative for the pathogens analyzed. There were no significant correlations between counts or count levels for thermotolerant coliforms or E. coli or the presence of F-RNA phages and pathogens in the samples analyzed.

  5. Endophytic Bacillus spp. produce antifungal lipopeptides and induce host defence gene expression in maize.

    PubMed

    Gond, Surendra K; Bergen, Marshall S; Torres, Mónica S; White, James F

    2015-03-01

    Endophytes are mutualistic symbionts within healthy plant tissues. In this study we isolated Bacillus spp. from seeds of several varieties of maize. Bacillus amyloliquifaciens or Bacillus subtilis were found to be present in all maize varieties examined in this study. To determine whether bacteria may produce antifungal compounds, generally lipopeptides in Bacillus spp., bacterial cultures were screened for production of lipopeptides. Lipopeptides were extracted by acid precipitation from liquid cultures of Bacillus spp. Lipopeptide extracts from Bacillus spp. isolated from Indian popcorn and yellow dent corn showed inhibitory activity against Fusarium moniliforme at 500μg per disk. Using MALDI-TOF mass spectrometry we detected the presence of antifungal iturin A, fengycin and bacillomycin in these isolates. PCR amplification also showed the presence of genes for iturin A and fengycin. B. subtilis (SG_JW.03) isolated from Indian popcorn showed strong inhibition of Arabidopsis seed mycoflora and enhanced seedling growth. We tested for the induction of defence gene expression in the host plant after treatment of plants with B. subtilis (SG_JW.03) and its lipopeptide extract using RT-qPCR. Roots of Indian popcorn seedlings treated with a suspension of B. subtilis (SG_JW.03) showed the induction of pathogenesis-related genes, including PR-1 and PR-4, which relate to plant defence against fungal pathogens. The lipopeptide extract alone did not increase the expression of these pathogenesis-related genes. Based on our study of maize endophytes, we hypothesize that, bacterial endophytes that naturally occur in many maize varieties may function to protect hosts by secreting antifungal lipopeptides that inhibit pathogens as well as inducing the up-regulation of pathogenesis-related genes of host plants (systemic acquired resistance). PMID:25497916

  6. Prevalence of Brucella spp in humans1

    PubMed Central

    Soares, Catharina de Paula Oliveira Cavalcanti; Teles, José Andreey Almeida; dos Santos, Aldenir Feitosa; Silva, Stemberg Oliveira Firmino; Cruz, Maria Vilma Rocha Andrade; da Silva-Júnior, Francisco Feliciano

    2015-01-01

    Objective: to determine the seroprevalence of Brucella spp in humans. Method: this is an observational study, developed with 455 individuals between 18 and 64 years old, who use the Estratégia de Saúde da Família (Brazil's family health strategy). The serum samples of volunteers underwent buffered acid antigen tests, such as screening, agar gel immunodiffusion and slow seroagglutination test in tubes and 2-Mercaptoethanol. Results: among the samples, 1.98% has responded to buffered-acid antigen, 2.85% to agar gel immunodiffusion test and 1.54% to the slow seroagglutination tests on tubes/2-Mercaptoethanol. The prevalence of Brucella spp was 4.4%, represented by the last two tests. Conclusion: the results of this research suggest that the studied population is exposed to Brucella spp infection. PMID:26487143

  7. Campylobacter spp. and birds of prey.

    PubMed

    Dipineto, Ludovico; De Luca Bossa, Luigi Maria; Russo, Tamara Pasqualina; Cutino, Eridania Annalisa; Gargiulo, Antonio; Ciccarelli, Francesca; Raia, Pasquale; Menna, Lucia Francesca; Fioretti, Alessandro

    2014-06-01

    A total of 170 birds of prey admitted to two Wildlife Rescue and Rehabilitation Centers of Italy were examined. Birds were divided by diurnal (n = 15) and nocturnal (n = 7) species, sampled by cloacal swabs, and examined for Campylobacter spp. by cultural and molecular methods. Campylobacter spp. were isolated in 43 out of the 170 (25.3%) birds of prey examined. Among these, 43/43 (100%) were identified as Campylobacter jejuni and 10/43 (23.3%) were identified as Campylobacter coli recovered from mixed infections. Diurnal birds of prey showed a significantly higher prevalence value (P = 0.0006) for Campylobacter spp. than did nocturnal birds of prey. PMID:25055637

  8. Cultural and Molecular Evidence of Legionella spp. Colonization in Dental Unit Waterlines: Which Is the Best Method for Risk Assessment?

    PubMed

    Ditommaso, Savina; Giacomuzzi, Monica; Ricciardi, Elisa; Zotti, Carla M

    2016-02-06

    Legionella spp. are ubiquitous in aquatic habitats and water distribution systems, including dental unit waterlines (DUWLs). The aim of the present study was to determine the prevalence of Legionella in DUWLs and tap water samples using PMA-qPCR and standard culture methods. The total viable counts (TVCs) of aerobic heterotrophic bacteria in the samples were also determined. Legionella spp. were detected and quantified using the modified ISO 11731 culture method. Extracted genomic DNA was analysed using the iQ-Check Quanti Legionella spp. kit, and the TVCs were determined according to the ISO protocol 6222. Legionella spp. were detected in 100% of the samples using the PMA-qPCR method, whereas these bacteria were detected in only 7% of the samples using the culture method. The number of colony forming units (CFUs) of the TVCs in the DUWL and tap water samples differed, with the bacterial load being significantly lower in the tap water samples (p-value = 0). The counts obtained were within the Italian standard range established for potable water in only 5% of the DUWL water samples and in 77% of the tap water samples. Our results show that the level of Legionella spp. contamination determined using the culture method does not reflect the true scale of the problem, and consequently we recommend testing for the presence of aerobic heterotrophic bacteria based on the assumption that Legionella spp. are components of biofilms.

  9. Cultural and Molecular Evidence of Legionella spp. Colonization in Dental Unit Waterlines: Which Is the Best Method for Risk Assessment?

    PubMed Central

    Ditommaso, Savina; Giacomuzzi, Monica; Ricciardi, Elisa; Zotti, Carla M.

    2016-01-01

    Legionella spp. are ubiquitous in aquatic habitats and water distribution systems, including dental unit waterlines (DUWLs). The aim of the present study was to determine the prevalence of Legionella in DUWLs and tap water samples using PMA-qPCR and standard culture methods. The total viable counts (TVCs) of aerobic heterotrophic bacteria in the samples were also determined. Legionella spp. were detected and quantified using the modified ISO 11731 culture method. Extracted genomic DNA was analysed using the iQ-Check Quanti Legionella spp. kit, and the TVCs were determined according to the ISO protocol 6222. Legionella spp. were detected in 100% of the samples using the PMA-qPCR method, whereas these bacteria were detected in only 7% of the samples using the culture method. The number of colony forming units (CFUs) of the TVCs in the DUWL and tap water samples differed, with the bacterial load being significantly lower in the tap water samples (p-value = 0). The counts obtained were within the Italian standard range established for potable water in only 5% of the DUWL water samples and in 77% of the tap water samples. Our results show that the level of Legionella spp. contamination determined using the culture method does not reflect the true scale of the problem, and consequently we recommend testing for the presence of aerobic heterotrophic bacteria based on the assumption that Legionella spp. are components of biofilms. PMID:26861373

  10. Quantitative detection of chloroethene-reductive bacteria Dehalococcoides spp. using alternately binding probe competitive Polymerase Chain Reaction.

    PubMed

    Miyata, Ryo; Adachi, Ken; Tani, Hidenori; Kurata, Shinya; Nakamura, Kazunori; Tsuneda, Satoshi; Sekiguchi, Yuji; Noda, Naohiro

    2010-06-01

    Dehalococcoides spp. are responsible for the reductive dehalogenation of environmental contaminants and are candidates for engineered bioremediation. The development of a sensitive, reliable, and rapid method for the quantification of Dehalococcoides spp. is required for the effective use of the organisms in bioremediation sites. Here, we describe the quantification of the 16S rRNA gene of Dehalococcoides spp. using a recently developed quantification method named alternately binding probe competitive PCR (ABC-PCR). The primers and probe sets that were newly designed for ABC-PCR were found to have a high specificity for Dehalococcoides spp. The standard curve of ABC-PCR had a good fitting (R = 0.999), and the lower detection limit was 10 copies/microl of template DNA. We also investigated the effects of inherent PCR-inhibiting compounds in an environmental sample on the quantification using ABC-PCR or real-time PCR by adding the soil extraction solution to PCR mixtures. ABC-PCR was more robust against the PCR amplification inhibitors than real-time PCR. The copy number of the 16S rRNA gene of Dehalococcoides spp. in soil and groundwater samples was successfully quantified using ABC-PCR. In conclusion, ABC-PCR is useful for the quantification of Dehalococcoides spp. populations and dynamics at bioremediation sites.

  11. The Antibiofilm Effect of Ginkgo biloba Extract Against Salmonella and Listeria Isolates from Poultry.

    PubMed

    Wu, Yan; Park, Keun Cheol; Choi, Beom Geun; Park, Jin Hwa; Yoon, Ki Sun

    2016-05-01

    Salmonella spp. and Listeria spp. are common foodborne pathogens in poultry and have caused a large number of outbreaks worldwide. Biofilm formation is common in the food industry and is also a mechanism of antimicrobial resistance. The aim of this work was to investigate the antimicrobial effect and mechanism of Ginkgo biloba extract against the biofilm formation of Salmonella and Listeria isolates from poultry at retail markets. Bacteria detection, isolation, and enumeration were carried out on 27 chicken and 29 ducks at retail markets. The effects of temperature and G. biloba extract against biofilm formation of Salmonella and Listeria isolates were measured using the crystal violet assay and swimming and swarming motilities. The monitoring results of Salmonella and Listeria in 56 poultry carcasses at retail markets in Korea showed that the prevalence of Salmonella spp. in poultry was low (5.4%), but the prevalence of Listeria spp (78.6%) was high. L. innocua was the predominant serotype (80%) in the isolated Listeria species. Temperature, strain, and surface affected the biofilm formation of Salmonella spp. and Listeria spp. L. innocua showed the best biofilm formation ability on a 96-well plate, while Salmonella Enteritidis formed the most biofilm on a glass slide. Biofilm formation abilities of Salmonella spp. and Listeria spp. were increased with the increase of temperature. G. biloba extract at 75 μg/mL significantly inhibited biofilm formation of Salmonella spp. and Listeria spp (p < 0.05). The mechanism of the antibiofilm effect of the G. biloba extract showed that the motility reduction may be one of the mechanisms of G. biloba extract against some serotypes of Salmonella and Listeria, but not L. monocytogenes. The findings of this study provided the basis for the application of G. biloba extract as a food additive to promote the quality and safety of poultry products.

  12. The Antibiofilm Effect of Ginkgo biloba Extract Against Salmonella and Listeria Isolates from Poultry.

    PubMed

    Wu, Yan; Park, Keun Cheol; Choi, Beom Geun; Park, Jin Hwa; Yoon, Ki Sun

    2016-05-01

    Salmonella spp. and Listeria spp. are common foodborne pathogens in poultry and have caused a large number of outbreaks worldwide. Biofilm formation is common in the food industry and is also a mechanism of antimicrobial resistance. The aim of this work was to investigate the antimicrobial effect and mechanism of Ginkgo biloba extract against the biofilm formation of Salmonella and Listeria isolates from poultry at retail markets. Bacteria detection, isolation, and enumeration were carried out on 27 chicken and 29 ducks at retail markets. The effects of temperature and G. biloba extract against biofilm formation of Salmonella and Listeria isolates were measured using the crystal violet assay and swimming and swarming motilities. The monitoring results of Salmonella and Listeria in 56 poultry carcasses at retail markets in Korea showed that the prevalence of Salmonella spp. in poultry was low (5.4%), but the prevalence of Listeria spp (78.6%) was high. L. innocua was the predominant serotype (80%) in the isolated Listeria species. Temperature, strain, and surface affected the biofilm formation of Salmonella spp. and Listeria spp. L. innocua showed the best biofilm formation ability on a 96-well plate, while Salmonella Enteritidis formed the most biofilm on a glass slide. Biofilm formation abilities of Salmonella spp. and Listeria spp. were increased with the increase of temperature. G. biloba extract at 75 μg/mL significantly inhibited biofilm formation of Salmonella spp. and Listeria spp (p < 0.05). The mechanism of the antibiofilm effect of the G. biloba extract showed that the motility reduction may be one of the mechanisms of G. biloba extract against some serotypes of Salmonella and Listeria, but not L. monocytogenes. The findings of this study provided the basis for the application of G. biloba extract as a food additive to promote the quality and safety of poultry products. PMID:26954614

  13. DNA microarray-based detection of multiple pathogens: Mycoplasma spp. and Chlamydia spp.

    PubMed

    Schnee, Christiane; Sachse, Konrad

    2015-01-01

    Rapid detection of slow-growing or non-culturable microorganisms, such as Mycoplasma spp. and Chlamydia spp., is still a challenge to diagnosticians in the veterinary field. In addition, as epidemiological evidence on the frequency of mixed infections involving two and more bacterial species has been emerging, detection methods allowing simultaneous identification of different pathogens are required. In the present chapter, we describe DNA microarray-based procedures for the detection of 83 Mollicutes species (Mycoplasma assay) and 11 Chlamydia spp. (Chlamydia assay). The assays are suitable for use in a routine diagnostic environment, as well as in microbiological research.

  14. 21 CFR 866.3740 - Streptococcus spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3740 Streptococcus spp. serological reagents. (a) Identification. Streptococcus spp. serological reagents are devices... streptococci are associated with infections, such as sore throat, impetigo (an infection characterized by...

  15. 21 CFR 866.3740 - Streptococcus spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3740 Streptococcus spp. serological reagents. (a) Identification. Streptococcus spp. serological reagents are devices... streptococci are associated with infections, such as sore throat, impetigo (an infection characterized by...

  16. HPLC determination of caffeine and theophylline in Paullinia cupana Kunth (guarana) and Cola spp. samples.

    PubMed

    Belliardo, F; Martelli, A; Valle, M G

    1985-05-01

    A reversed-phase high-performance liquid-chromatographic method for the determination of caffeine and theophylline in commercial guarana samples (drug obtained from the seeds of Paulinia cupana Kunth, Sapindaceae of the Amazon Region) and in Cola spp. samples is described and discussed. The methodology developed is simple and rapid with a minimum of samples preparation required. A comparison of five different techniques for the extraction of caffeine and theophylline is discussed. Furthermore the quantitative determination of caffeine and theophylline in five samples of Brasilian guarana, in two samples of dietetic products containing guarana, in two samples of Cola extract and in three of Cola seed powder are reported.

  17. Malassezia spp. overgrowth in allergic cats.

    PubMed

    Ordeix, Laura; Galeotti, Franca; Scarampella, Fabia; Dedola, Carla; Bardagí, Mar; Romano, Erica; Fondati, Alessandra

    2007-10-01

    A series of 18 allergic cats with multifocal Malassezia spp. overgrowth is reported: atopic dermatitis was diagnosed in 16, an adverse food reaction in another and one was euthanized 2 months after diagnosis of Malassezia overgrowth. All the cats were otherwise healthy and those tested (16 out of 18) for feline leukaemia or feline immunodeficiency virus infections were all negative. At dermatological examination, multifocal alopecia, erythema, crusting and greasy adherent brownish scales were variably distributed on all cats. Cytological examination revealed Malassezia spp. overgrowth with/without bacterial infection in facial skin (n = 11), ventral neck (n = 6), abdomen (n = 6), ear canal (n = 4), chin (n = 2), ear pinnae (n = 2), interdigital (n = 1) and claw folds skin (n = 1). Moreover, in two cats Malassezia pachydermatis was isolated in fungal cultures from lesional skin. Azoles therapy alone was prescribed in seven, azoles and antibacterial therapy in eight and azoles with both antibacterial and anti-inflammatory therapy in three of the cats. After 3-4 weeks of treatment, substantial reduction of pruritus and skin lesions was observed in all 11 cats treated with a combined therapy and in five of seven treated solely with azoles. Malassezia spp. overgrowth may represent a secondary cutaneous problem in allergic cats particularly in those presented for dermatological examination displaying greasy adherent brownish scales. The favourable response to treatment with antifungal treatments alone suggests that, as in dogs, Malassezia spp. may be partly responsible for both pruritus and cutaneous lesions in allergic cats. PMID:17845619

  18. Evaluating SPP/APR Improvement Activities

    ERIC Educational Resources Information Center

    National Early Childhood Technical Assistance Center (NECTAC), 2009

    2009-01-01

    This document is intended to assist State Education Agency (SEA) and Lead Agency (LA) staff and technical assistance providers in designing a meaningful evaluation for the State Performance Plan (SPP)/Annual Performance Report (APR) improvement activities. It provides: (1) information about the relevance of evaluation in the context of improvement…

  19. Characterization of Milkweed (Asclepias spp.) Seed Proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Milkweed (Asclepias spp.) is a crop grown mainly for the production of floss used as hypoallergenic fillers in comforters and pillows. The seeds end up as by-products. Milkweed seed contains 21% oil and 30% crude protein (dry basis). The oil is similar in quality to soybean oil, but there is no i...

  20. Biological control of saltcedar (Tamarix spp.) by saltcedar leaf beetles (Diorhabda spp.): effects on small mammals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The spread of introduced saltcedar (Tamarix spp.) throughout many riparian systems across the western United States motivated the introduction of biological control agents that are specific to saltcedar, saltcedar leaf beetles (Diorhabda carinulata, D. elongata; Chrysomelidae). I monitored small mam...

  1. Suppressive effects of metabolites from Photorhabdus spp. and Xenorhabdus spp. on phytopathogens of peach and pecan

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Our objective was to determine the suppressive abilities of bacterial metabolites derived from Photorhabdus and Xenorhabdus spp. on Glomerella cingulata, Phomopsis sp., Phytophthora cactorum, and Fusicladosporium effusum, which are fungal or oomycete pathogens of pecan, and Monilinia fructicola, a f...

  2. Molecular identification of Sarcocystis spp. in foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) from Germany.

    PubMed

    Moré, G; Maksimov, A; Conraths, F J; Schares, G

    2016-04-15

    More than 200 Sarcocystis spp. have been named and most of them appear to be involved in a particular predator-prey cycle. Among canids, the European fox (Vulpes vulpes) and the raccoon dog (Nyctereutes procyonoides) are widely distributed in Europe and probably play an important role as definitive hosts in the epidemiology of Sarcocystis spp. infections. A total of 50 small intestines from foxes and 38 from raccoon dogs were sampled in the Federal State of Brandenburg, Germany. Mucosal scrapings were collected and analyzed by sugar flotation and when oocysts or sporocysts were detected, an overnight sedimentation was performed and DNA extracted with a commercial kit. A PCR was conducted using primers targeting a fragment of the 18S rRNA gene (with a size of approximately 850 bp) and the amplicons were purified and sequenced. Samples with an inconclusive sequencing were cloned into plasmids and ≥ 3 plasmids from each amplicon were sequenced. Sarcocystis spp. oocysts/sporocysts were detected in 38% (19/50) of fox and 52.6% (20/38) of raccoon dog samples. Sequencing analysis of amplicons from oocyst DNA revealed mixed infections in 9 fox and 5 raccoon dog samples. In the fox samples, the most often identified Sarcocystis spp. were S. tenella or S. capracanis (10.0%); S. miescheriana (8.0%) and S. gracilis (8.0%) followed by Sarcocystis spp., which use birds as intermediate hosts (6.0%), and S. capreolicanis (4.0%). In the raccoon dog samples, sequences with a ≥99% identity with the following species were detected: S. miescheriana (18.4%), S. gracilis (13.1%), Sarcocystis spp. using birds as IH (10.5%), S. tenella or S.capracanis (2.6%) and S. capreolicanis (2.6%). The estimated prevalence of Sarcocystis spp. infections determined using mucosal scrapings was higher than in related studies performed by analyzing faecal samples. The methodology of 18S rRNA gene amplification, cloning and sequencing is suitable to identify mixed infections with Sarcocystis spp. and

  3. Molecular identification of Sarcocystis spp. in foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) from Germany.

    PubMed

    Moré, G; Maksimov, A; Conraths, F J; Schares, G

    2016-04-15

    More than 200 Sarcocystis spp. have been named and most of them appear to be involved in a particular predator-prey cycle. Among canids, the European fox (Vulpes vulpes) and the raccoon dog (Nyctereutes procyonoides) are widely distributed in Europe and probably play an important role as definitive hosts in the epidemiology of Sarcocystis spp. infections. A total of 50 small intestines from foxes and 38 from raccoon dogs were sampled in the Federal State of Brandenburg, Germany. Mucosal scrapings were collected and analyzed by sugar flotation and when oocysts or sporocysts were detected, an overnight sedimentation was performed and DNA extracted with a commercial kit. A PCR was conducted using primers targeting a fragment of the 18S rRNA gene (with a size of approximately 850 bp) and the amplicons were purified and sequenced. Samples with an inconclusive sequencing were cloned into plasmids and ≥ 3 plasmids from each amplicon were sequenced. Sarcocystis spp. oocysts/sporocysts were detected in 38% (19/50) of fox and 52.6% (20/38) of raccoon dog samples. Sequencing analysis of amplicons from oocyst DNA revealed mixed infections in 9 fox and 5 raccoon dog samples. In the fox samples, the most often identified Sarcocystis spp. were S. tenella or S. capracanis (10.0%); S. miescheriana (8.0%) and S. gracilis (8.0%) followed by Sarcocystis spp., which use birds as intermediate hosts (6.0%), and S. capreolicanis (4.0%). In the raccoon dog samples, sequences with a ≥99% identity with the following species were detected: S. miescheriana (18.4%), S. gracilis (13.1%), Sarcocystis spp. using birds as IH (10.5%), S. tenella or S.capracanis (2.6%) and S. capreolicanis (2.6%). The estimated prevalence of Sarcocystis spp. infections determined using mucosal scrapings was higher than in related studies performed by analyzing faecal samples. The methodology of 18S rRNA gene amplification, cloning and sequencing is suitable to identify mixed infections with Sarcocystis spp. and

  4. Maternal antibodies against Plasmodium spp. in African black-footed penguin (Spheniscus demersus) chicks.

    PubMed

    Graczyk, T K; Cranfield, M R; Shaw, M L; Craig, L E

    1994-07-01

    Anti-Plasmodium spp. antibody titers of mating pairs of adult, captive-reared, African black-footed penguins (Spheniscus demersus) and their chicks were determined using the enzyme-linked-immunosorbent assay (ELISA). Two Plasmodium falciparum antigens were used for the ELISA: R32tet32 (sporozoite antigen), and crude red blood cell extract (CRBCE). Eighteen chicks were bled weekly for ten weeks starting with their day of hatching. The yolk sacs of two penguin eggs were biopsed for ELISA-detectable maternal antibodies (MAB). None of the 28 adult penguins were parasitemic by Giemsa-stained thin blood smear; however, all had anti-Plasmodium spp. immunoglobulins reacting with P. falciparum antigens. All 18 newly hatched chicks had anti-Plasmodium spp. MAB while housed in a mosquito-free environment. The level of MAB in the newly hatched chicks was correlated significantly (P < 0.001) with antibody level detected in their female parents (R32tet32: r = 0.87, CRBCE: r = 0.89). No correlation was found between antibody titers of the newly hatched chicks and their male parents. The level of maternal-fetal antibodies was regressed significantly (P < 0.001) over the 10-week period. Penguin chicks over 10 weeks of age had no anti-Plasmodium spp. MAB. Egg-yolk samples had significantly (P < 0.03) higher MAB titers than female parents that laid these eggs.

  5. Isolation of amoebae and Pseudomonas and Legionella spp. from eyewash stations.

    PubMed Central

    Paszko-Kolva, C; Yamamoto, H; Shahamat, M; Sawyer, T K; Morris, G; Colwell, R R

    1991-01-01

    Forty eyewash units were sampled for protozoa, bacteria, and fungi. Total heterotrophic bacterial counts on nutrient agar and R2A agar (Difco Laboratories, Detroit, Mich.) ranged from 0 to 10(5) CFU/ml, with Pseudomonas spp. being the most frequently isolated. Total counts of 10(4) and 10(8) cells per ml were obtained with the acridine orange staining procedure. All samples were examined for Legionella spp. by direct fluorescent-antibody staining and by culturing on buffered charcoal-yeast extract agar containing alpha-ketoglutarate and glycine and supplemented with cycloheximide, vancomycin, and polymyxin B. DNA-DNA hybridization was used to confirm identification of the Legionella isolates. Legionellae were detected in 35 of 40 (87.5%) samples by direct fluorescent-antibody staining, with 3 samples yielding both Legionella spp. and amoebae. Amoebae identified as Hartmannella, Vahlkampfia, Acanthamoeba, and Cochliopodium spp. were detected in 19 of 40 (47:5%) samples. Sabouraud dextrose agar was used to obtain a crude estimate of viable fungal populations, pH, hardness, and ammonia, alkalinity, chlorine, copper, and iron contents were recorded for all water samples collected from eyewash stations; 33% of the samples had greater than or equal to 10 mg of CO2 per liter. It is concluded that eyewash stations not regularly flushed and/or cleaned and used to flush traumatized eye tissue may be a source of infection and can contaminate laboratory environments via aerosol transmission. PMID:2036003

  6. Polyphosphate-degrading enzymes in Acinetobacter spp. and activated sludge.

    PubMed Central

    van Groenestijn, J W; Bentvelsen, M M; Deinema, M H; Zehnder, A J

    1989-01-01

    Polyphosphate-degrading enzymes were studied in Acinetobacter spp. and activated sludge. Polyphosphate: AMP phosphotransferase activity in Acinetobacter strain 210A decreased with increasing growth rates. The activity of this enzyme in cell extracts of Acinetobacter strain 210A was maximal at a pH of 8.5 and a temperature of 40 degrees C and was stimulated by (NH4)2SO4. The Km for AMP was 0.6 mM, and the Vmax was 60 nmol/min per mg of protein. Cell extracts of this strain also contained polyphosphatase, which was able to degrade native polyphosphate and synthetic magnesium polyphosphate and was strongly stimulated by 300 to 400 mM NH4Cl. A positive correlation was found between polyphosphate:AMP phosphotransferase activity, adenylate kinase activity, and phosphorus accumulation in six Acinetobacter strains. Significant activities of polyphosphate kinase were detected only in strain P, which contained no polyphosphate:AMP phosphotransferase. In samples of activated sludge from different plants, the activity of adenylate kinase correlated well with the ability of the sludge to remove phosphate biologically from wastewater. PMID:2539774

  7. The Role of Malassezia spp. in Atopic Dermatitis.

    PubMed

    Glatz, Martin; Bosshard, Philipp P; Hoetzenecker, Wolfram; Schmid-Grendelmeier, Peter

    2015-01-01

    Malassezia spp. is a genus of lipophilic yeasts and comprises the most common fungi on healthy human skin. Despite its role as a commensal on healthy human skin, Malassezia spp. is attributed a pathogenic role in atopic dermatitis. The mechanisms by which Malassezia spp. may contribute to the pathogenesis of atopic dermatitis are not fully understood. Here, we review the latest findings on the pathogenetic role of Malassezia spp. in atopic dermatitis (AD). For example, Malassezia spp. produces a variety of immunogenic proteins that elicit the production of specific IgE antibodies and may induce the release of pro-inflammatory cytokines. In addition, Malassezia spp. induces auto-reactive T cells that cross-react between fungal proteins and their human counterparts. These mechanisms contribute to skin inflammation in atopic dermatitis and therefore influence the course of this disorder. Finally, we discuss the possible benefit of an anti-Malassezia spp. treatment in patients with atopic dermatitis. PMID:26239555

  8. The Role of Malassezia spp. in Atopic Dermatitis

    PubMed Central

    Glatz, Martin; Bosshard, Philipp P.; Hoetzenecker, Wolfram; Schmid-Grendelmeier, Peter

    2015-01-01

    Malassezia spp. is a genus of lipophilic yeasts and comprises the most common fungi on healthy human skin. Despite its role as a commensal on healthy human skin, Malassezia spp. is attributed a pathogenic role in atopic dermatitis. The mechanisms by which Malassezia spp. may contribute to the pathogenesis of atopic dermatitis are not fully understood. Here, we review the latest findings on the pathogenetic role of Malassezia spp. in atopic dermatitis (AD). For example, Malassezia spp. produces a variety of immunogenic proteins that elicit the production of specific IgE antibodies and may induce the release of pro-inflammatory cytokines. In addition, Malassezia spp. induces auto-reactive T cells that cross-react between fungal proteins and their human counterparts. These mechanisms contribute to skin inflammation in atopic dermatitis and therefore influence the course of this disorder. Finally, we discuss the possible benefit of an anti-Malassezia spp. treatment in patients with atopic dermatitis. PMID:26239555

  9. Actinomyces spp. gene expression in root caries lesions

    PubMed Central

    Dame-Teixeira, Naile; Parolo, Clarissa Cavalcanti Fatturi; Maltz, Marisa; Tugnait, Aradhna; Devine, Deirdre; Do, Thuy

    2016-01-01

    Background The studies of the distribution of Actinomyces spp. on carious and non-carious root surfaces have not been able to confirm the association of these bacteria with root caries, although they were extensively implicated as a prime suspect in root caries. Objective The aim of this study was to observe the gene expression of Actinomyces spp. in the microbiota of root surfaces with and without caries. Design The oral biofilms from exposed sound root surface (SRS; n=10) and active root caries (RC; n=30) samples were collected. The total bacterial RNA was extracted, and the mRNA was isolated. Samples with low RNA concentration were pooled, yielding a final sample size of SRS=10 and RC=9. Complementary DNA (cDNA) libraries were prepared and sequenced on an Illumina® HiSeq 2500 system. Sequence reads were mapped to eight Actinomyces genomes. Count data were normalized using DESeq2 to analyse differential gene expression applying the Benjamini-Hochberg correction (false discovery rate [FDR]<0.001). Results Actinomyces spp. had similar numbers of reads (Mann-Whitney U-test; p>0.05), except for Actinomyces OT178 (p=0.001) and Actinomyces gerencseriae (p=0.004), which had higher read counts in the SRS. Genes that code for stress proteins (clp, dnaK, and groEL), enzymes of glycolysis pathways (including enolase and phosphoenolpyruvate carboxykinase), adhesion (Type-2 fimbrial and collagen-binding protein), and cell growth (EF-Tu) were highly – but not differentially (p>0.001) – expressed in both groups. Genes with the most significant upregulation in RC were those coding for hypothetical proteins and uracil DNA glycosylase (p=2.61E-17). The gene with the most significant upregulation in SRS was a peptide ABC transporter substrate-binding protein (log2FC=−6.00, FDR=2.37E-05). Conclusion There were similar levels of Actinomyces gene expression in both sound and carious root biofilms. These bacteria can be commensal in root surface sites but may be cariogenic

  10. Prevalence of Bartonella spp. in Canine Cutaneous Histiocytoma.

    PubMed

    Pultorak, E L; Linder, K; Maggi, R G; Balakrishnan, N; Breitschwerdt, E B

    2015-07-01

    Canine cutaneous histiocytoma (CCH) is a common, benign neoplastic proliferation of histiocytes of Langerhans cell origin that often ulcerate, become secondarily infected and regress spontaneously. Bartonella is a fastidious genus of facultative intracellular pathogens that can be transmitted through arthropod bites and epidermal animal scratches and has been identified previously in the cytoplasm of histiocytes within granulomatous lesions and in skin biopsy samples of inflammatory pustules and papules. Based on the established inflammatory and oncogenic properties of Bartonella, we hypothesized that Bartonella spp. DNA could be amplified from CCH more often than from non-lesional skin and bacteria could be localized within skin tumours using indirect immunofluorescence (IIF). Paraffin wax-embedded surgical biopsy samples from dogs with CCH and non-neoplastic skin adjacent to osteosarcomas (control group selected due to wide surgical margins) were retrieved from the archive of the pathology service of North Carolina State University College of Veterinary Medicine. DNA was extracted and regions of the 16S-23S rRNA intergenic transcribed spacer (ITS) region and the pap31 and gltA genes were amplified by polymerase chain reaction (PCR) using Bartonella-specific primers. IIF was performed using a primary Bartonella henselae monoclonal antibody to localize B. henselae in tissues of PCR-positive dogs. Bartonella vinsonii subsp. berkhoffii was amplified from 1/17 (5.8%) control tissues and B. henselae was amplified from 4/29 (13.8%) CCH tissues. The prevalence of B. vinsonii subsp. berkhoffii (P = 0.37) or B. henselae (P = 0.28) did not vary statistically between study groups. B. henselae could be visualized in 2/4 (50.0%) CCH tissues using IIF. Based on this study, Bartonella spp. are unlikely to cause CCH. PMID:25980841

  11. Occurrence of Potentially Pathogenic Bacterial-Endosymbionts in Acanthamoeba Spp.

    PubMed Central

    NIYYATI, Maryam; MAFI, Mahyar; HAGHIGHI, Ali; HAKEMI VALA, Mojdeh

    2015-01-01

    Background: Acanthamoeba- bacteria interactions enable pathogenic bacteria to tolerate harsh conditions and lead to transmission to the susceptible host. The present study was aimed to address the presence of bacterial endosymbionts of Acanthamoeba isolated from recreational water sources of Tehran, Iran. To the best of our knowledge this is the first study regarding occurrence of bacteria in environmental Acanthamoeba spp. in Iran. Methods: A total of 75 samples of recreational water sources were collected. Samples were cultured on non- nutrient agar 1.5% plates. Positive Acanthamoeba spp. were axenically grown. DNA extraction and PCR reaction was performed using JDP1-2 primers. All positive samples of Acanthamoeba were examined for the presence of endosymbionts using staining and molecular methods. The PCR products were then sequenced in order to determine the genotypes of Acanthamoeba and bacteria genera. Results: Out of 75 samples, 16 (21.3%) plates were positive for Acanthamoeba according to the morphological criteria. Molecular analysis revealed that Acanthamoeba belonged to T4 and T5 genotypes. Five isolates (35.7%) were positive for bacterial endosymbionts using staining method and PCR test. Sequencing of PCR products confirmed the presence of Pseudomonas aeruginosa and Agrobacterium tumefasiens. Conclusion: The presence of Acanthamoeba bearing pathogenic endosymbionts in water sources leads us to public health issues including improved sanitation and decontamination measures in recreational water sources in order to prevent amoebae-related infection. To the best of our knowledge this is the first report regarding the isolation of A. tumefasiens from Acanthamoeba in Iran and worldwide. PMID:26246815

  12. Serological and molecular investigation of Ehrlichia spp. and Anaplasma spp. in ticks and blood of dogs, in the Thrace Region of Turkey.

    PubMed

    Çetinkaya, Handan; Matur, Erdal; Akyazi, İbrahim; Ekiz, Elif Ergul; Aydin, Levent; Toparlak, Mufit

    2016-07-01

    In recent years, tick-borne diseases like ehrlichiosis and anaplasmosis became widespread worldwide threatening the health of both human and companion animals. Therefore, the aim of this study was to determine the presence of Anaplasma spp., and Ehrlichia spp. in dogs and ticks in the Thrace Region of Turkey. A total of 400 blood samples and 912 ticks were collected from dogs living in shelters that are located in four cities (Istanbul, Edirne, Tekirdag and Kirklareli) of the Thrace Region. Blood and buffy coat smears were prepared for microscopic examination. Hematologic and serologic analyses were performed using cell counter and commercial Snap3Dx test kit, respectively. Eight hundred fifty of collected ticks were classified as Rhipicephalus sanguineus, 33 as Rhipicephalus turanicus and 29 as Ixodes ricinus. After DNA extraction from blood samples and pooled ticks (127 tick pools, in total), nested PCR was performed to detect the DNA of Anaplasma spp., and Ehrlichia spp. The seroprevalence of Ehrlichia canis was 27.25% (109) by Snap3Dx test and the total molecular positivity was 11.75% (47) in dog blood samples and 21.25% (27) in tick pools by nested PCR. The frequencies of the infected blood samples with E. canis, Anaplasma phagocytophilum and Anaplasma platys were detected as 6%, 4% and 6%, respectively. E. canis and A. platys were detected in R. sanguineus pools with a ratio of 15.75% and 0.7%, respectively. In addition, A. platys was also detected in R. turanicus pools (0.7%). A. phagocytophilum was found only in I. ricinus pools (3.93%). Morulae of three species were detected in buffy coat and blood smears. While anemia was observed in dogs infected with E. canis and co-infected (with one or more species), thrombocytopenia was observed only in co-infected dogs. This is the first study providing evidence for the presence of Anaplasma spp. and Ehrlichia spp. in dogs and ticks in the Thrace Region of Turkey. Based on the results of the tests used in this study

  13. Serological and molecular investigation of Ehrlichia spp. and Anaplasma spp. in ticks and blood of dogs, in the Thrace Region of Turkey.

    PubMed

    Çetinkaya, Handan; Matur, Erdal; Akyazi, İbrahim; Ekiz, Elif Ergul; Aydin, Levent; Toparlak, Mufit

    2016-07-01

    In recent years, tick-borne diseases like ehrlichiosis and anaplasmosis became widespread worldwide threatening the health of both human and companion animals. Therefore, the aim of this study was to determine the presence of Anaplasma spp., and Ehrlichia spp. in dogs and ticks in the Thrace Region of Turkey. A total of 400 blood samples and 912 ticks were collected from dogs living in shelters that are located in four cities (Istanbul, Edirne, Tekirdag and Kirklareli) of the Thrace Region. Blood and buffy coat smears were prepared for microscopic examination. Hematologic and serologic analyses were performed using cell counter and commercial Snap3Dx test kit, respectively. Eight hundred fifty of collected ticks were classified as Rhipicephalus sanguineus, 33 as Rhipicephalus turanicus and 29 as Ixodes ricinus. After DNA extraction from blood samples and pooled ticks (127 tick pools, in total), nested PCR was performed to detect the DNA of Anaplasma spp., and Ehrlichia spp. The seroprevalence of Ehrlichia canis was 27.25% (109) by Snap3Dx test and the total molecular positivity was 11.75% (47) in dog blood samples and 21.25% (27) in tick pools by nested PCR. The frequencies of the infected blood samples with E. canis, Anaplasma phagocytophilum and Anaplasma platys were detected as 6%, 4% and 6%, respectively. E. canis and A. platys were detected in R. sanguineus pools with a ratio of 15.75% and 0.7%, respectively. In addition, A. platys was also detected in R. turanicus pools (0.7%). A. phagocytophilum was found only in I. ricinus pools (3.93%). Morulae of three species were detected in buffy coat and blood smears. While anemia was observed in dogs infected with E. canis and co-infected (with one or more species), thrombocytopenia was observed only in co-infected dogs. This is the first study providing evidence for the presence of Anaplasma spp. and Ehrlichia spp. in dogs and ticks in the Thrace Region of Turkey. Based on the results of the tests used in this study

  14. Genotyping of Giardia lamblia and Entamoeba spp from river waters in Iran.

    PubMed

    Mahmoudi, Mohammad Reza; Nazemalhosseini-Mojarad, Ehsan; Karanis, Panagiotis

    2015-12-01

    In this study, DNA from 55 surface and river water samples, which were collected from some water sources of Tehran and the Guilan Province, Iran, were extracted and examined for Entamoeba spp. and Giardia lamblia by PCR and genotyping. Twenty-seven samples, which were concentrated using the immunomagnetic separation technology (IMS) method, were examined for Giardia alone. Twenty-eight samples, which were concentrated using the sucrose flotation (SF) method, were examined for both Giardia and Entamoeba species. The results showed that 27/55 (17/27 and 10/28) (49 %), 4 /28 (14.28 %) and 3/28 (10.7 %) of the samples were positive for Giardia lamblia, Entamoeba spp and mixed infections (Entamoeba spp. and Giardia spp.), respectively. Sixteen out of 55 samples were negative. Entamoeba genus-specific PCR primers in single-round PCR were used to differentiate between the Entamoeba spp. (E. histolytica, E. dispar and E. moshkovskii). With respect to the 7 samples that were positive for Entamoeba, (14.28 %) 4 out of 28 were positive for E. moshkovskii, (7.14 %), 2 out of 28 were positive for E. histolytica and (3.57 %) 1 out of 28 was positive for E. dispar. Genus-specific PCR primers in a semi-nested PCR assay was performed to genotype Giardia species. Of the 27 samples that were positive for Giardia, 10 samples were sequences. All 10 successfully sequenced samples contained assemblage B of Giardia lamblia.This is first study to investigate the G. lamblia genotypes in the water supply of the Tehran and Guilan provinces, and it is the first study to investigate Entamoeba species in the water supplies of Iran. The investigated river water supplies, which are used for agriculture, camping and animal farming, were heavily contaminated by the human pathogenic Entamoeba and Giardia parasites. There is a potential risk of waterborne outbreaks in humans and animals. PMID:26350378

  15. Trichophyton Spp. fungal keratitis in 22 years old female contact lenses wearer.

    PubMed

    Mravicić, Ivana; Dekaris, Iva; Gabrić, Nikica; Romac, Ivana; Glavota, Vlade; Sviben, Mario

    2010-04-01

    Fungal keratitis represents one of the most difficult forms of microbial keratitis to diagnose and treat successfully. It is difficult to obtain correct diagnosis and topical antifungal preparations. Fungi can cause severe stromal necrosis and enter the anterior chamber by penetrating an intact Descemet membrane. The most common pathogens are filamentous fungi (Aspergillus and Fusarium spp.) and Candida albicans. The incidence of Trichophyton spp. keratitis is 5%. A 22 years old female contact lenses wearer after keratitis developed corneal melting syndrome, spontaneous perforation of the cornea and complicated cataract of the left eye. Conjunctival swab was sterile as well as first sample of corneal tissue and sample from the anterior chamber. Urgent therapeutic perforating keratoplasty (PK), was performed together with extracapsular cataract extraction and the implantation of the intraocular lens in the posterior chamber. The patient was treated with ciprofloxacin and diflucan (systemic therapy); with dexamethason and atropin (subconjunctivaly) and chlorhexidine, brolene, levofloxacin, polimyxin B, and dexamethason/neomycin (topically). Microbiology evaluation was performed once again following excisional biopsy of the intracameral portion of the lesion. The presence of Trichophyton spp. was finally confirmed. Itraconazole and garamycin were included in the systemic therapy. Corneal graft was clear for 17 days but decompensated 28 days after the PK. After two weeks microorganisms invaded the vitreous and caused endophthalmitis. Despite urgent pars plana vitrectomy patient developed endophthalmitis, lost light sensation and developed phthysis. Evisceration and the implantation of silicon prosthesis was done. Perforating keratoplasty is a method of choice in treating severe infectious keratitis unresponsive to conservative treatment but without the eradication of microorganisms it cannot restore the vision or save the eye. Trichophyton spp. may cause a severe

  16. Genotyping of Giardia lamblia and Entamoeba spp from river waters in Iran.

    PubMed

    Mahmoudi, Mohammad Reza; Nazemalhosseini-Mojarad, Ehsan; Karanis, Panagiotis

    2015-12-01

    In this study, DNA from 55 surface and river water samples, which were collected from some water sources of Tehran and the Guilan Province, Iran, were extracted and examined for Entamoeba spp. and Giardia lamblia by PCR and genotyping. Twenty-seven samples, which were concentrated using the immunomagnetic separation technology (IMS) method, were examined for Giardia alone. Twenty-eight samples, which were concentrated using the sucrose flotation (SF) method, were examined for both Giardia and Entamoeba species. The results showed that 27/55 (17/27 and 10/28) (49 %), 4 /28 (14.28 %) and 3/28 (10.7 %) of the samples were positive for Giardia lamblia, Entamoeba spp and mixed infections (Entamoeba spp. and Giardia spp.), respectively. Sixteen out of 55 samples were negative. Entamoeba genus-specific PCR primers in single-round PCR were used to differentiate between the Entamoeba spp. (E. histolytica, E. dispar and E. moshkovskii). With respect to the 7 samples that were positive for Entamoeba, (14.28 %) 4 out of 28 were positive for E. moshkovskii, (7.14 %), 2 out of 28 were positive for E. histolytica and (3.57 %) 1 out of 28 was positive for E. dispar. Genus-specific PCR primers in a semi-nested PCR assay was performed to genotype Giardia species. Of the 27 samples that were positive for Giardia, 10 samples were sequences. All 10 successfully sequenced samples contained assemblage B of Giardia lamblia.This is first study to investigate the G. lamblia genotypes in the water supply of the Tehran and Guilan provinces, and it is the first study to investigate Entamoeba species in the water supplies of Iran. The investigated river water supplies, which are used for agriculture, camping and animal farming, were heavily contaminated by the human pathogenic Entamoeba and Giardia parasites. There is a potential risk of waterborne outbreaks in humans and animals.

  17. Easy storage strategies for Sporothrix spp. strains.

    PubMed

    Brilhante, Raimunda Sâmia Nogueira; Silva, Natalya Fechine; Lima, Rita Amanda Chaves de; Caetano, Érica Pacheco; Alencar, Lucas Pereira de; Castelo-Branco, Débora de Souza Collares Maia; Moreira, José Luciano Bezerra; Bandeira, Silviane Praciano; Camargo, Zoilo Pires de; Rodrigues, Anderson Messias; Bandeira, Tereza de Jesus Pinheiro Gomes; Monteiro, André Jalles; Cordeiro, Rossana de Aguiar; Sidrim, José Júlio Costa; Rocha, Marcos Fábio Gadelha

    2015-04-01

    The present study evaluated the maintenance of Sporothrix spp. (6 Sporothrix brasiliensis; 6 S. schenckii; 5 S. mexicana, and 3 S. globosa) in saline at 4°C, and in 10% glycerol plus either 10% lactose or 10% sucrose, at -20°C and -80°C. Viability was assessed after 3, 6, and 9 months of storage, through the recovery of strains on potato dextrose agar and analysis of macro- and micromorphological features. Conidium quantification was performed before and after storage, at 3, 6 and 9 months. 100% viability was observed, regardless of storage conditions or time period. Storage at 4°C and at -20°C did not alter the number of conidia, but lower conidium counts were observed at -80°C. This study shows that the combination of glycerol with lactose or sucrose is effective to maintain Sporothrix spp. at freezing temperatures.

  18. Bartonella spp. in Small Mammals, Benin.

    PubMed

    Martin-Alonso, Aarón; Houemenou, Gualbert; Abreu-Yanes, Estefanía; Valladares, Basilio; Feliu, Carlos; Foronda, Pilar

    2016-04-01

    This study aimed to investigate the prevalence and genetic diversity of Bartonella organisms in small mammals in Cotonou, Benin. We captured 163 rodents and 12 insectivores and successfully detected Bartonella DNA from 13 of the 175 small mammal individuals. Bartonella spp., identical or closely related to Bartonella elizabethae, Bartonella tribocorum, and Bartonella rochalimae, was detected. A potential new Bartonella species, proposed as Candidatus Bartonella mastomydis, was found in three Mastomys individuals and genetically characterized by targeting two housekeeping genes (rpoB and gltA) and the intergenic species region. However, 20.8% of gray rats were found to be infected with Bartonella spp., and none of the black rats analyzed was positive. This work may be important from a public health point of view due to the zoonotic nature of the Bartonella species detected and warrants further investigation on the unknown zoonotic potential of this newly proposed Bartonella species. PMID:26910412

  19. Bartonella spp. in Small Mammals, Benin.

    PubMed

    Martin-Alonso, Aarón; Houemenou, Gualbert; Abreu-Yanes, Estefanía; Valladares, Basilio; Feliu, Carlos; Foronda, Pilar

    2016-04-01

    This study aimed to investigate the prevalence and genetic diversity of Bartonella organisms in small mammals in Cotonou, Benin. We captured 163 rodents and 12 insectivores and successfully detected Bartonella DNA from 13 of the 175 small mammal individuals. Bartonella spp., identical or closely related to Bartonella elizabethae, Bartonella tribocorum, and Bartonella rochalimae, was detected. A potential new Bartonella species, proposed as Candidatus Bartonella mastomydis, was found in three Mastomys individuals and genetically characterized by targeting two housekeeping genes (rpoB and gltA) and the intergenic species region. However, 20.8% of gray rats were found to be infected with Bartonella spp., and none of the black rats analyzed was positive. This work may be important from a public health point of view due to the zoonotic nature of the Bartonella species detected and warrants further investigation on the unknown zoonotic potential of this newly proposed Bartonella species.

  20. Blood cell findings resembling Bartonella spp.

    PubMed

    Pitassi, Luiza Helena Urso; Cintra, Maria Letícia; Ferreira, Marilucia Ruggiero Martins; Magalhães, Renata Ferreira; Velho, Paulo Eduardo Neves Ferreira

    2010-02-01

    Some Bartonella species are able to invade red blood cells (RBC) and may cause persistent infection in the susceptible host. Use of transmission electron microscopy (TEM) demonstrates, inside erythrocytes, the typical triple-walled agents. However, when examining ultrathin sections of blood cells, the authors have, on several occasions, detected intraerythrocytic abnormalities that mimic but are not typical of Bartonella spp. Small endovesicles, pseudoinclusions, cavities, and irregular hemoglobin granules distribution, resulting in regions of increased or decreased electron density, may be observed in the erythrocytes and platelets, which may be confused with bartonellas. So far, detailed ultrastructural findings of Bartonella spp. in blood cells have not yet been described. Aiming to improve TEM interpretation of blood cells changes, in routine examination of blood sections of patients with suspected bartonellosis, the authors studied the morphological findings they have observed, and present their putative nature, according to information in the literature.

  1. Mysterious chronic urticaria caused by Blastocystis spp.?

    PubMed

    Lepczyńska, Małgorzata; Chen, Wen-Chieh; Dzika, Ewa

    2016-03-01

    Species of the genus Blastocystis, which are single-cell, intestinal protozoan parasites of humans and animals, remain mysterious, with unclear clinical and epidemiologic significance. In recent years, many researchers have suggested a possible connection between Blastocystis spp. infection and chronic urticaria. In the present article, we review the literature and discuss the possible associations between the clinical symptomatology and pathogenicity of this organism in terms of its subtypes, morphologic forms, genetic diversity, and interactions with other intestinal microbiota. PMID:26469206

  2. Non-lethal detection of DNA from Cichlidogyrus spp. (Monogenea, Ancyrocephalinae) in gill mucus of the Nile tilapia Oreochromis niloticus.

    PubMed

    Ek-Huchim, Juan Pablo; Jimenez-Garcia, Isabel; Pérez-Vega, Juan Antonio; Rodríguez-Canul, Rossanna

    2012-03-20

    Infection of Nile tilapia Oreochromis niloticus by monogeneans of the genus Cichlidogyrus is harmful. Currently, diagnosis of this infection is based on invasive techniques and the identification of isolated parasites by their morphology. To facilitate diagnosis, we have developed a non-lethal polymerase chain reaction (PCR) test for detection of Cichlidogyrus spp. DNA in the gill mucus of O. niloticus, using 5 pairs of specific primers based on Cichlidogyrus sclerosus 28S rRNA (Cicly 1 to Cicly 5) which generate fragments of approximately 188, 180, 150, 159 and 189 bp, respectively. PCR specificity was tested using genomic DNA extracted individually from 175 isolated Cichlidogyrus spp., 75 Gyrodactylus cichlidarum and 75 endopararasitic Enterogyrus spp., as well as from 75 protozoans Trichodina spp. The Cicly primers were used to detect Cichlidogyrus spp. DNA in mucus from the gills of 23 Nile tilapia confirmed to be infected with the parasite. Negative controls consisted of 45 uninfected Nile tilapia. The limit of sensitivity of the assay was 1.2 ng of purified parasite DNA. The Cicly primers did not amplify DNA from the mucus of non-infected Nile tilapia, G. cichlidarum, Trichodina spp. or Enterogyrus spp. In all cases, the sensitivity and specificity of the test were 100%. The sequences of all the amplified fragments showed a high similarity to that of the 28S rRNA region of C. sclerosus (93 to 100% identical to GenBank Accession No. DQ157660.1). We provide evidence for a safe and non-invasive DNA-based diagnostic method for the presence of Cichlidogyrus in the gill mucus of O. niloticus. PMID:22436463

  3. Isolation and Quantification of Pinitol, a Bioactive Cyclitol, in Retama spp.

    PubMed

    González-Mauraza, Nuria H; León-González, Antonio J; Espartero, José L; Gallego-Fernández, Juan B; Sánchez-Hidalgo, Marina; Martin-Cordero, Carmen

    2016-03-01

    The genus Retama (Fabaceae) is widely distributed in the Mediterranean region. In the present study, pinitol (3-O-methyl-chiro-inositol), an anti-inflammatory and antidiabetic molecule, was isolated from aerial parts of R. monosperma, and its structure established on the basis of spectroscopic techniques (1D/2D NMR) and MS. Identification and quantification of pinitol in R. raetam and R. sphaerocarpa were also performed. R. monosperma had the highest concentration of pinitol (2.3%). The presence of pinitol in aqueous extracts of Retama spp. may explain the adaptation of these plants to drought and salinity. Furthermore, pinitol could be considered as a mediator in the anti-inflammatory and hypoglycemic activities of Retama spp., which are traditionally used to treat diabetes. PMID:27169192

  4. Micropropagation of Rubus and Ribes spp.

    PubMed

    Dziedzic, Ewa; Jagła, Joanna

    2013-01-01

    Micropropagation is the most appropriate method for large-scale production of Rubus and Ribes spp. The proliferation rate of Rubus spp. differs in shoot tips and nodal segments. The culture media used for raspberry and blackberry propagation are MS-based supplemented with different combination and ratio of plant growth regulators, depending on the stage of culture. The initiation medium containing 0.4 mg L(-1) BA and 0.1 mg L(-1) IBA is used to stabilize shoot cultures. In multiplication media, concentration of cytokinin is doubled. In vitro rooting of shoots is achieved on media supplemented with 1.0 mg L(-1) IBA. Ribes spp. cultures are initiated from shoot tips, meristem, or dormant buds on MS medium supplemented with 2.0 mg L(-1) BA, 0.5 mg L(-1) IBA, and 0.1 mg L(-1) GA(3.) After stabilization of shoot cultures in 3-4-week time, shoot multiplication is carried out on MS medium containing 1.0 mg L(-1) BA and 0.1 mg L(-1) IBA. Shoots 2 cm long are cultured to rooting on a medium amended with 2.0 mg L(-1) IBA and 5.0 mg L(-1) IAA. Rooted plantlets are transferred to universal peat substrate and acclimatized in the greenhouse.

  5. Enrichment of Acinetobacter spp. from food samples.

    PubMed

    Carvalheira, Ana; Ferreira, Vânia; Silva, Joana; Teixeira, Paula

    2016-05-01

    Relatively little is known about the role of foods in the chain of transmission of acinetobacters and the occurrence of different Acinetobacter spp. in foods. Currently, there is no standard procedure to recover acinetobacters from food in order to gain insight into the food-related ecology and epidemiology of acinetobacters. This study aimed to assess whether enrichment in Dijkshoorn enrichment medium followed by plating in CHROMagar™ Acinetobacter medium is a useful method for the isolation of Acinetobacter spp. from foods. Recovery of six Acinetobacter species from food spiked with these organisms was compared for two selective enrichment media (Baumann's enrichment and Dijkshoorn's enrichment). Significantly (p < 0.01) higher cell counts were obtained in Dijkshoorn's enrichment. Next, the Dijkshoorn's enrichment followed by direct plating on CHROMagar™ Acinetobacter was applied to detect Acinetobacter spp. in different foods. Fourteen different presumptive acinetobacters were recovered and assumed to represent nine different strains on the basis of REP-PCR typing. Eight of these strains were identified by rpoB gene analysis as belonging to the species Acinetobacter johnsonii, Acinetobacter calcoaceticus, Acinetobacter guillouiae and Acinetobacter gandensis. It was not possible to identify the species level of one strain which may suggests that it represents a distinct species.

  6. The occurrence of Listeria spp. and Salmonella spp. in surface waters.

    PubMed

    Arvanitidou, M; Papa, A; Constantinidis, T C; Danielides, V; Katsouyannopoulos, V

    1997-12-01

    Listeria ssp., mainly Listeria monocytogenes as well as Salmonella spp. are recognized as significant human pathogens. The purpose of this study was to examine the occurrence of Listeria spp. and Salmonella spp. in surface waters of Northern Greece and to investigate the correlation of these pathogens with the standard indicator bacteria. A total number of 128 water samples from four rivers and one lake were examined for the presence of Listeria, Salmonella, total coliforms, faecal coliforms and faecal streptococci. For isolating Listeria, 250 ml of water were filtered through 0.45 microns pore size membrane, that was transferred in 10 ml listeria enrichment broth and after incubation for 24 h at 30 degrees C, a second enrichment in FDA and Fraser broths was followed. After 24 hour incubation, an amount of 0.1 ml was streaked out onto listeria selective medium. The typical colonies were further biochemically and serologically examined. Salmonella spp. were isolated after preenrichment in BPW, enrichment in Rappaport-Vassiliadis and selenite cysteine broths and identified from BGD and SS agar plates by biochemistry and serology. Listeria monocytogenes was isolated from five (3.9%) and Salmonella spp. from eight (6.2%) samples. Mean log values of the standard indicator bacteria did not significantly differ between listeria and salmonella positive and negative samples.

  7. Flea species infesting dogs in Florida and Bartonella spp. prevalence rates.

    PubMed

    Yore, K; DiGangi, B; Brewer, M; Balakrishnan, N; Breitschwerdt, E B; Lappin, M

    2014-01-31

    Several Bartonella spp. associated with fleas can induce a variety of clinical syndromes in both dogs and humans. However, few studies have investigated the prevalence of Bartonella in the blood of dogs and their fleas. The objectives of this study were to determine the genera of fleas infesting shelter dogs in Florida, the prevalence of Bartonella spp. within the fleas, and the prevalence of Bartonella spp. within the blood of healthy dogs from which the fleas were collected. Fleas, serum, and EDTA-anti-coagulated whole blood were collected from 80 healthy dogs, and total DNA was extracted for PCR amplification of Bartonella spp. The genera of fleas infesting 43 of the dogs were determined phenotypically. PCR amplicons from blood and flea pools were sequenced to confirm the Bartonella species. Amplicons for which sequencing revealed homology to Bartonella vinsonii subsp. berkhoffii (Bvb) underwent specific genotyping by targeting the 16S-23S intergenic spacer region. A total of 220 fleas were collected from 80 dogs and pooled by genus (43 dogs) and flea species. Bartonella spp. DNA was amplified from 14 of 80 dog blood samples (17.5%) and from 9 of 80 pooled fleas (11.3%). B. vinsonii subsp. berkhoffii DNA was amplified from nine dogs and five of the flea pools. Bartonella rochalimae (Br) DNA was amplified from six dogs and two flea pools. One of 14 dogs was co-infected with Bvb and Br. The dog was infested with Pulex spp. fleas containing Br DNA and a single Ctenocephalides felis flea. Of the Bvb bacteremic dogs, five and four were infected with genotypes II and I, respectively. Of the Bvb PCR positive flea pools, three were Bvb genotype II and two were Bvb genotype I. Amplification of Bvb DNA from Pulex spp. collected from domestic dogs, suggests that Pulex fleas may be a vector for dogs and a source for zoonotic transfer of this pathogen from dogs to people. The findings of this study provide evidence to support the hypothesis that flea-infested dogs may be a

  8. Development of Rapid Isothermal Amplification Assays for Detection of Phytophthora spp. in Plant Tissue.

    PubMed

    Miles, Timothy D; Martin, Frank N; Coffey, Michael D

    2015-02-01

    Several isothermal amplification techniques recently have been developed that are tolerant of inhibitors present in many plant extracts, which can reduce the need for obtaining purified DNA for running diagnostic assays. One such commercially available technique that has similarities with real-time polymerase chain reaction (PCR) for designing primers and a labeled probe is recombinase polymerase amplification (RPA). This technology was used to develop two simple and rapid approaches for detection of Phytophthora spp.: one genus-specific assay multiplexed with a plant internal control and the other species-specific assays for Phytophthora ramorum and P. kernoviae. All assays were tested for sensitivity (ranging from 3 ng to 1 fg of DNA) and specificity using DNA extracted from more than 136 Phytophthora taxa, 21 Pythium spp., 1 Phytopythium sp., and a wide range of plant species. The lower limit of linear detection using purified DNA was 200 to 300 fg of DNA in all pathogen RPA assays. Six different extraction buffers were tested for use during plant tissue maceration and the assays were validated in the field by collecting 222 symptomatic plant samples from over 50 different hosts. Only 56 samples were culture positive for Phytophthora spp. whereas 91 were positive using the Phytophthora genus-specific RPA test and a TaqMan real-time PCR assay. A technique for the generation of sequencing templates from positive RPA amplifications to confirm species identification was also developed. These RPA assays have added benefits over traditional technologies because they are rapid (results can be obtained in as little as 15 min), do not require DNA extraction or extensive training to complete, use less expensive portable equipment than PCR-based assays, and are significantly more specific than current immunologically based methods. This should provide a rapid, field-deployable capability for pathogen detection that will facilitate point-of-sample collection processing

  9. A new species of Eurytoma (Hymenoptera: Eurytomidae) attacking, Quadrastichus spp. (Hymenoptera: Eulophidae) galling Erythrina spp. (Fabaceae) with a summary of African Eurytoma spp. biology and species checklist

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eurytoma erythrinae Gates and Delvare, new species, is described and illustrated. This species was reared from field-collected galls induced on Erythrina spp. by Quadrastichus spp. (Hymenoptera: Eulophidae), in Tanzania, Ghana, and South Africa. It is compared to a closely related African species. W...

  10. Evaluation of the relatedness of Brucella spp. and Ochrobactrum anthropi and description of Ochrobactrum intermedium sp. nov., a new species with a closer relationship to Brucella spp.

    PubMed

    Velasco, J; Romero, C; López-Goñi, I; Leiva, J; Díaz, R; Moriyón, I

    1998-07-01

    The relatedness of Brucella spp. and Ochrobactrum anthropi was studied by protein profiling, Western blot, immunoelectrophoresis and 16S rRNA analysis. Whole-cell and soluble proteins of brucellae and O. anthropi showed serological cross-reactivities quantitatively and qualitatively more intense than those existing with similar extracts of Agrobacterium spp. Numerical analysis of Western blot profiles of whole-cell extracts showed that O. anthropi LMG 3301 was closer to Brucella spp. than to O. anthropi LMG 3331T, a result not obtained by protein profiling. These differences were not observed by Western blot with soluble fractions, and immunoelectrophoretic analyses suggested that this was due to destruction of conformational epitopes in Western blot procedures with the subsequent simplification of antigenic profile. Analysis of the 16S rRNA sequences of strains previously used in the species definition confirmed that strain LMG 3301, and also LMG 3306, were closer to the brucellae, and that LMG 3331T was in a separate cluster. The LMG 3301 and the LMG 3331T clusters could also be separated by their different colistin sensitivity and by PCR with 16S rRNA Brucella primers, and both methods showed strains of both clusters among clinical isolates classified as O. anthropi by conventional tests. These results and those of previous DNA-DNA hybridization studies [Holmes, B., Popoff, M., Kiredjian, M. & Kersters, K. (1988). Int J Syst Bacteriol 38, 406-416] show that the LMG 3301 cluster and related clinical isolates should be given a new species status for which the name Ochrobactrum intermedium sp. nov. is proposed (type strain is LMG 3301T=NCTC 12171T = CNS 2-75T).

  11. Chemical Components and Cardiovascular Activities of Valeriana spp.

    PubMed Central

    Chen, Heng-Wen; Wei, Ben-Jun; He, Xuan-Hui; Liu, Yan; Wang, Jie

    2015-01-01

    Valeriana spp. is a flowering plant that is well known for its essential oils, iridoid compounds such as monoterpenes and sesquiterpenes, flavonoids, alkaloids, amino acids, and lignanoids. Valeriana spp. exhibits a wide range of biological activities such as lowering blood pressure and heart rate, antimyocardial ischemia reperfusion injury, antiarrhythmia, and regulation of blood lipid levels. This review focuses on the chemical constituents and cardiovascular activities of Valeriana spp. PMID:26788113

  12. Transpiration rates of rice plants treated with Trichoderma spp.

    NASA Astrophysics Data System (ADS)

    Doni, Febri; Anizan, I.; Che Radziah C. M., Z.; Yusoff, Wan Mohtar Wan

    2014-09-01

    Trichoderma spp. are considered as successful plant growth promoting fungi and have positive role in habitat engineering. In this study, the potential for Trichoderma spp. to regulate transpiration process in rice plant was assessed experimentally under greenhouse condition using a completely randomized design. The study revealed that Trichoderma spp. have potential to enhance growth of rice plant through transpirational processes. The results of the study add to the advancement of the understanding as to the role of Trichoderma spp. in improving rice physiological process.

  13. Medicinal leech therapy and Aeromonas spp. infection.

    PubMed

    Verriere, B; Sabatier, B; Carbonnelle, E; Mainardi, J L; Prognon, P; Whitaker, I; Lantieri, L; Hivelin, M

    2016-06-01

    While the use of medicinal leech therapy (MLT) in reconstructive and orthopaedic surgery is widely described, post-operative complications related to leeches remain a major concern. Aeromonas spp. strains are involved in the majority of reported cases. As surgical success rate is directly impacted, an adapted antibiotic prophylaxis should be instituted in order to minimize these complications. We assessed pharmaceutical process, microbiological control and related infections in order to provide data and choose the appropriate antibiotherapy for patients requiring MLT. We report a clinical and microbiological study over a 24-month period. Clinical data were collected from patients' database, and microbiological analysis both on leeches' tank water and crushed leeches were performed to characterize isolated strains and their susceptibility to antibiotics. A total of 595 leeches were used to treat 28 patients (12 in plastic surgery and 16 in orthopaedic surgery), and three documented cases of post-operative infections were reported. Aeromonas spp. isolates yielded from 62 % of analyzed batches (75 % of Aeromonas veronii). Eighteen Aeromonas spp. isolates yielded from 23 water samples and three crushed leeches. Isolates were similar in tank and crushed leeches. Strains were susceptible to fluoroquinolones, sulfamethoxazole/trimethoprim, aminosides, and third-generation cephalosporins but resistant to amoxicillin/clavulanic acid and second-generation cephalosporins. According to collected data, routine tank water microbiological analyses are mandatory in order to identify leeches' batches containing resistant strains and to discard them. In this context, the surgeon is able to select an appropriated antibiotic prophylaxis in order to avoid MLT associated serious post-operative complications.

  14. Inhibition of Quorum Sensing in Staphylococcus spp.

    PubMed

    Brackman, Gilles; Coenye, Tom

    2015-01-01

    The Gram-positive, facultative anaerobic coccus-shaped bacteria of the genus Staphylococcus are among the most important causative agents of acute and chronic bacterial infections in humans as well as in animals. Treatment of Staphylococcus infections has become increasingly challenging due to the growing problem of antibiotic resistance. For this reason innovative antimicrobials with novel targets and modes of action are needed. Since the discovery that QS is used by Staphylococcus spp. to coordinate the expression of several genes involved in virulence, biofilm formation and pathogenicity, QS inhibition has gained increasing attention as an alternative anti-pathogenic strategy. A major advantage compared with antibiotic therapy is that QSIs are used in concentrations that do not affect bacterial growth. For this reason, it is expected that these compounds would exert less pressure towards the development of resistance. However, some important points still need to be addressed. Although several inhibitors have proven to be active antipathogenic agents in vitro and in several in vivo models, it is still unknown whether these compounds will also be useful in humans. Furthermore, several fundamental mechanisms by which the different QS systems in Staphylococcus spp. exert their regulatory functions and how they are inhibited by QSIs are still poorly understood. In order to achieve real-life applications with QSIs, these challenges should be addressed and more research will be needed. In this article, we will discuss the different QS systems present in Staphylococcus spp., how they are used to control virulence and biofilm formation and how they can be blocked.

  15. Eimeria spp. in Brazilian water buffalo.

    PubMed

    de Noronha, Antonio Carlos F; Starke-Buzetti, Wilma A; Duszynski, Donald W

    2009-02-01

    Eimeria species are frequently found in water buffalo (Bubalus bubalis) in Brazil. Here, we report those Eimeria spp. that infect buffalos during their first year of life. Fresh fecal samples were examined from 2 groups (1 group/yr for 2 yr, 2000-2002), each with 18 water buffalo calves (both sexes), from birth through 12 mo of age, in Selvíria, MS, Brazil. Five oocyst morphotypes were observed, i.e., Eimeria ellipsoidalis and Eimeria zuernii, both previously described from water buffalo, and 3 other morphotypes consistent with descriptions of known Eimeria spp. from Artiodactyla hosts, but originally described from other genera than those in which we found them (referred to here as Eimeria species 1-3). Our results showed that buffalo calves started shedding oocysts in their feces between 6-29 days of age, with the highest concentration ranging from 188-292 oocysts/g of feces. The 3 unnamed oocyst morphotypes in the calf feces resembled E. auburnensis (Eimeria sp. 3), E. cylindrica (Eimeria sp. 1), and E. subspherica (Eimeria sp. 2). The most prevalent species were Eimeria sp. 1 and E. ellipsoidalis, which dominated in the youngest animals (6 to 133 days old). Eimeria zuernii oocysts, in contrast, were found only in low numbers in the feces of older calves (208 to 283 days old). Calves were infected more frequently during the rainy season (September to January) in both years, but cows were negative for Eimeria spp., whenever feces were collected (spring, winter, autumn, or summer seasons).

  16. Isolation of Yersinia spp. from bovine feces.

    PubMed

    Fukushima, H; Saito, K; Tsubokura, M; Otsuki, K; Kawaoka, Y

    1983-10-01

    Yersinia spp. were sought in 618 fecal samples from cows. Four strains of Yersinia enterocolitica, serotype O:12,26 (one), O:13,7 (two), and O:18 (one); seven strains of Yersinia kristensenii, serotype O:11,24 (five) and O:12,26 (two); and one strain of Yersinia pseudotuberculosis serotype IIB, were isolated. This is the first time that Y. pseudotuberculosis has been isolated from cows in Japan, and the isolation of serotype IIB of this organism from cows seems to be the first in the world.

  17. Anther culture of chili pepper (Capsicum spp.).

    PubMed

    Ochoa-Alejo, Neftalí

    2012-01-01

    Chili pepper (Capsicum spp.) is a very important horticultural crop around the world and is especially important for Mexicans because of its impact in the culture and the cuisine. Biotechnological tools such as tissue culture techniques and specifically anther culture may be applied successfully for plant breeding and genetic improvement in order to generate isogenic lines (100% homozygous) in a shorter time in comparison with the classic breeding methods. In this chapter, a protocol for efficient recovery of chili pepper haploid plants from in vitro cultured anthers is described. PMID:22610631

  18. Endemic Viruses of Squirrel Monkeys (Saimiri spp.)

    PubMed Central

    Rogers, Donna L; McClure, Gloria B; Ruiz, Julio C; Abee, Christian R; Vanchiere, John A

    2015-01-01

    Nonhuman primates are the experimental animals of choice for the study of many human diseases. As such, it is important to understand that endemic viruses of primates can potentially affect the design, methods, and results of biomedical studies designed to model human disease. Here we review the viruses known to be endemic in squirrel monkeys (Saimiri spp.). The pathogenic potential of these viruses in squirrel monkeys that undergo experimental manipulation remains largely unexplored but may have implications regarding the use of squirrel monkeys in biomedical research. PMID:26141448

  19. [Hypersensitivity pneumonitis after exposure to Candida spp].

    PubMed

    Serrano, Carlos; Torrego, Alfonso; Loosli, Alfonso; Valero, Antonio; Picado, César

    2010-05-01

    Hypersensitivity pneumonitis (HP) is a lung disease caused by heavy and recurrent inhalation of antigens. We describe the case of a patient with HP caused by domestic exposure to Candida spp. The diagnosis was made by taking into consideration the, clinical presentation, exposure history, radiological findings, bronchoalveolar lavage, lung function and the immuno-allergy study. The diagnosis was definitively confirmed by performing a specific bronchial provocation test. It has been shown that there is cross-reactivity between different Candida species, and despite making the diagnosis in this case with Candida albicans, we were unable to define exactly which species was responsible for the HP.

  20. Volatile composition screening of Salix spp. nectar honey: benzenecarboxylic acids, norisoprenoids, terpenes, and others.

    PubMed

    Jerković, Igor; Marijanović, Zvonimir

    2010-09-01

    Salix spp. nectar honey volatiles of Croatian origin were analyzed by headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE), followed by gas chromatography and mass spectrometry (GC, GC/MS). Isolated volatiles were found in the honey headspace and extracts with almost exclusive distribution of several abundant compounds (e.g., phenylacetic acid, pinocembrin, 8-hydroxy-4,7-dimethylcoumarin, and 3-hydroxy-trans-β-damascone in the extracts, or safranal and lilac alcohols in the headspace). Comparison with Croatian Salix spp. honeydew honey revealed similarities regarding distribution of important shikimate pathway derivatives (e.g., high percentage of phenylacetic acid) and several norisoprenoids (α-isophorone and 4-oxoisophorone). On the other hand, distinct features of this honey were occurrence of compounds such as pinocembrin, 8-hydroxy-4,7-dimethylcoumarin, phenylacetonitrile, norisoprenoids (major ones: 3-hydroxy-trans-β-damascenone and trans-β-damascone), more pronounced variability of linalool-derived compounds, as well as the abundance of 3-methylpropanoic acid, 3-methylbutanoic acid, 2-methylpentanoic acid, and 3-methylpentan-1-ol.

  1. Characterization of geographically distinct bacterial communities associated with coral mucus produced by Acropora spp. and Porites spp.

    PubMed

    McKew, B A; Dumbrell, A J; Daud, S D; Hepburn, L; Thorpe, E; Mogensen, L; Whitby, C

    2012-08-01

    Acropora and Porites corals are important reef builders in the Indo-Pacific and Caribbean. Bacteria associated with mucus produced by Porites spp. and Acropora spp. from Caribbean (Punta Maroma, Mexico) and Indo-Pacific (Hoga and Sampela, Indonesia) reefs were determined. Analysis of pyrosequencing libraries showed that bacterial communities from Caribbean corals were significantly more diverse (H', 3.18 to 4.25) than their Indonesian counterparts (H', 2.54 to 3.25). Dominant taxa were Gammaproteobacteria, Alphaproteobacteria, Firmicutes, and Cyanobacteria, which varied in relative abundance between coral genera and region. Distinct coral host-specific communities were also found; for example, Clostridiales were dominant on Acropora spp. (at Hoga and the Mexican Caribbean) compared to Porites spp. and seawater. Within the Gammproteobacteria, Halomonas spp. dominated sequence libraries from Porites spp. (49%) and Acropora spp. (5.6%) from the Mexican Caribbean, compared to the corresponding Indonesian coral libraries (<2%). Interestingly, with the exception of Porites spp. from the Mexican Caribbean, there was also a ubiquity of Psychrobacter spp., which dominated Acropora and Porites libraries from Indonesia and Acropora libraries from the Caribbean. In conclusion, there was a dominance of Halomonas spp. (associated with Acropora and Porites [Mexican Caribbean]), Firmicutes (associated with Acropora [Mexican Caribbean] and with Acropora and Porites [Hoga]), and Cyanobacteria (associated with Acropora and Porites [Hoga] and Porites [Sampela]). This is also the first report describing geographically distinct Psychrobacter spp. associated with coral mucus. In addition, the predominance of Clostridiales associated with Acropora spp. provided additional evidence for coral host-specific microorganisms.

  2. Characterization of Geographically Distinct Bacterial Communities Associated with Coral Mucus Produced by Acropora spp. and Porites spp.

    PubMed Central

    McKew, B. A.; Dumbrell, A. J.; Daud, S. D.; Hepburn, L.; Thorpe, E.; Mogensen, L.

    2012-01-01

    Acropora and Porites corals are important reef builders in the Indo-Pacific and Caribbean. Bacteria associated with mucus produced by Porites spp. and Acropora spp. from Caribbean (Punta Maroma, Mexico) and Indo-Pacific (Hoga and Sampela, Indonesia) reefs were determined. Analysis of pyrosequencing libraries showed that bacterial communities from Caribbean corals were significantly more diverse (H′, 3.18 to 4.25) than their Indonesian counterparts (H′, 2.54 to 3.25). Dominant taxa were Gammaproteobacteria, Alphaproteobacteria, Firmicutes, and Cyanobacteria, which varied in relative abundance between coral genera and region. Distinct coral host-specific communities were also found; for example, Clostridiales were dominant on Acropora spp. (at Hoga and the Mexican Caribbean) compared to Porites spp. and seawater. Within the Gammproteobacteria, Halomonas spp. dominated sequence libraries from Porites spp. (49%) and Acropora spp. (5.6%) from the Mexican Caribbean, compared to the corresponding Indonesian coral libraries (<2%). Interestingly, with the exception of Porites spp. from the Mexican Caribbean, there was also a ubiquity of Psychrobacter spp., which dominated Acropora and Porites libraries from Indonesia and Acropora libraries from the Caribbean. In conclusion, there was a dominance of Halomonas spp. (associated with Acropora and Porites [Mexican Caribbean]), Firmicutes (associated with Acropora [Mexican Caribbean] and with Acropora and Porites [Hoga]), and Cyanobacteria (associated with Acropora and Porites [Hoga] and Porites [Sampela]). This is also the first report describing geographically distinct Psychrobacter spp. associated with coral mucus. In addition, the predominance of Clostridiales associated with Acropora spp. provided additional evidence for coral host-specific microorganisms. PMID:22636010

  3. Evidence of protocarnivory in triggerplants (Stylidium spp.; Stylidiaceae).

    PubMed

    Darnowski, D W; Carroll, D M; Płachno, B; Kabanoff, E; Cinnamon, E

    2006-11-01

    Australian triggerplants (Stylidium spp.; Stylidiaceae) trap small insects using mucilage-secreting glandular hairs held at various points on their inflorescence stems and flower parts. Triggerplants are generally found in habitats also containing genera of plants already accepted as carnivorous, two of which (Drosera, Byblis) use the same basic mechanism as Stylidium to trap their prey. In the herbarium, sheets of triggerplants and of accepted groups of carnivorous plants held similar numbers of trapped insects, and in the field, trapping of small prey per unit of glandular surface area was the same at a given site for triggerplants and for nearby carnivorous plants at three sites in northern Australia. Even more important, protease activity was produced by glandular regions of both triggerplants and Drosera after induction with yeast extract. A panel of negative and positive controls, including use 1) of plants grown in tissue culture, which therefore lack surface microorganisms, and 2) of protease inhibitors, shows that this activity 1) is generated by the glandular regions of the triggerplant itself, not by organisms that might reside on the surface of the plants, and 2) is due to proteases. All of this evidence taken together provides strong evidence of protocarnivory in Stylidium, something not previously suggested in the scientific literature, though the insect trapping has been noted informally. Experiments remain to be done to determine nutrient uptake, so triggerplants may well be fully carnivorous. PMID:17058181

  4. Avian wildlife reservoir of Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. in Norway.

    PubMed

    Kapperud, G; Rosef, O

    1983-02-01

    Cloacal swabs from 540 wild-living birds were cultured for Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. The carrier rates detected were as follows: C. fetus subsp. jejuni, 28.4%; Yersinia spp., 1.2%; and Salmonella spp., 0.8%. All birds were apparently healthy when captured. C. fetus subsp. jejuni was isolated from 11 of the 40 bird species examined. Among birds inhabiting the city of Oslo, the highest isolation rate was found in crows (Corvus corone cornix) (89.8%), followed by gulls (Larus spp.) (50.0%) and domestic pigeons (Columba livia domesticus) (4.2%). The gulls and crows scavenge on refuse dumps. High carrier rates were also detected among the following birds from nonurban, coastal areas: puffin (Fratercula arctica) (51.3%), common tern (Sterna hirundo) (5.6%), common gull (Larus canus) (18.9%), black-headed gull (Larus ridibundus) (13.2%), and herring gull (Larus argentatus) (4.2%). The list of species harboring C. fetus subsp. jejuni also includes the Ural owl (Strix uralensis), goldeneye (Bucephala clangula), and reed bunting (Emberiza schoeniclus). The following five Yersinia strains were isolated: Y. kristensenii (two strains), Y. intermedia (two strains), and "Yersinia X2" (one strain). Four strains belonging to the genus Salmonella were isolated from three different species of gulls. These isolates were identified as S. typhimurium, S. indiana, and S. djugu. The results indicate that campylobacters are a normal component of the intestinal flora in several bird species, whereas Salmonella and Yersinia carriers are more sporadic. PMID:6338824

  5. Avian wildlife reservoir of Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. in Norway.

    PubMed

    Kapperud, G; Rosef, O

    1983-02-01

    Cloacal swabs from 540 wild-living birds were cultured for Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. The carrier rates detected were as follows: C. fetus subsp. jejuni, 28.4%; Yersinia spp., 1.2%; and Salmonella spp., 0.8%. All birds were apparently healthy when captured. C. fetus subsp. jejuni was isolated from 11 of the 40 bird species examined. Among birds inhabiting the city of Oslo, the highest isolation rate was found in crows (Corvus corone cornix) (89.8%), followed by gulls (Larus spp.) (50.0%) and domestic pigeons (Columba livia domesticus) (4.2%). The gulls and crows scavenge on refuse dumps. High carrier rates were also detected among the following birds from nonurban, coastal areas: puffin (Fratercula arctica) (51.3%), common tern (Sterna hirundo) (5.6%), common gull (Larus canus) (18.9%), black-headed gull (Larus ridibundus) (13.2%), and herring gull (Larus argentatus) (4.2%). The list of species harboring C. fetus subsp. jejuni also includes the Ural owl (Strix uralensis), goldeneye (Bucephala clangula), and reed bunting (Emberiza schoeniclus). The following five Yersinia strains were isolated: Y. kristensenii (two strains), Y. intermedia (two strains), and "Yersinia X2" (one strain). Four strains belonging to the genus Salmonella were isolated from three different species of gulls. These isolates were identified as S. typhimurium, S. indiana, and S. djugu. The results indicate that campylobacters are a normal component of the intestinal flora in several bird species, whereas Salmonella and Yersinia carriers are more sporadic.

  6. DIFFERENTIATION BETWEEN Nocardia spp. AND Mycobacterium spp.: CRITICAL ASPECTS FOR BACTERIOLOGICAL DIAGNOSIS

    PubMed Central

    Muricy, Edna Cleide Mendes; Lemes, Romilda Aparecida; Bombarda, Sidney; Ferrazoli, Lucilaine; Chimara, Erica

    2014-01-01

    New methodologies were developed for the identification of Nocardia but the initial diagnosis still requires a fast and accurate method, mainly due to the similarity to Mycobacterium, both clinical and bacteriologically. Growth on Löwenstein-Jensen (LJ) medium, presence of acid-fast bacilli through Ziehl-Neelsen staining, and colony morphology can be confusing aspects between Nocardia and Mycobacterium. This study describes the occurrence of Nocardia spp. in a mycobacterial-reference laboratory, observing the main difficulties in differentiating Nocardia spp. from Mycobacterium spp., and correlating isolates with nocardiosis cases. Laboratory records for the period between 2008 and 2012 were analyzed, and the isolates identified as Nocardia sp. or as non-acid-fast filamentous bacilli were selected. Epidemiological and bacteriological data were analyzed as well. Thirty-three isolates identified as Nocardia sp. and 22 as non-acid-fast bacilli were selected for this study, and represented 0.12% of isolates during the study period. The presumptive identification was based on macroscopic and microscopic morphology, resistance to lysozyme and restriction profiles using the PRA-hsp65 method. Nocardia spp. can grow on media for mycobacteria isolation (LJ and BBL MGIT™) and microscopy and colony morphology are very similar to some mycobacteria species. Seventeen patients (54.8%) were reported and treated for tuberculosis, but presented signs and symptoms of nocardiosis. It was concluded that the occurrence of Nocardia sp. during the study period was 0.12%. Isolates with characteristics of filamentous bacilli, forming aerial hyphae, with colonies that may be pigmented, rough and without the BstEII digestion pattern in PRA-hsp65 method are suggestive of Nocardia spp. For a mycobacterial routine laboratory, a flow for the presumptive identification of Nocardia is essential, allowing the use of more accurate techniques for the correct identification, proper treatment and

  7. Single-step PCR for detection of Brucella spp. from blood and milk of infected animals.

    PubMed Central

    Leal-Klevezas, D S; Martínez-Vázquez, I O; López-Merino, A; Martínez-Soriano, J P

    1995-01-01

    A versatile method for the extraction of Brucella DNA and PCR are presented as reliable tools for the detection of Brucella spp. from body fluids of infected animals. Two oligonucleotides homologous to regions of the gene encoding for an outer membrane protein (omp-2) were designed to detect the pathogen from milk and/or blood of infected goats, bovines, and human patients. The sensitivity of our test and its ability to detect the pathogen in samples from the field reveal a promising advance in the diagnosis of brucellosis in animals and humans. PMID:8586678

  8. An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

    USGS Publications Warehouse

    Starliper, Clifford E.; Ketolab, Henry G.; Noyes, Andrew D.; Schill, William B.; Henson, Fred G.; Chalupnicki, Marc A.; Dittman, Dawn E.

    2015-01-01

    Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments for captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine if selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC’s) were determined for these four oils, Allimed® (garlic extract, Allium sativum) and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBC’s (0.02 to 0.04%) were obtained with three different sources of cinnamon oil. MBC’s for three sources of oregano and lemongrass oils ranged from 0.14 to 0.30% and 0.10 to 0.65%, respectively, and for two thyme oils were 2.11 and 2.22%. The highest concentration (5%) of Allimed® tested resulted in MBC’s to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBC’s for all but one isolate

  9. An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

    PubMed

    Starliper, Clifford E; Ketola, Henry G; Noyes, Andrew D; Schill, William B; Henson, Fred G; Chalupnicki, Marc A; Dittman, Dawn E

    2015-01-01

    Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments of captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine whether selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC's) were determined for these four oils, Allimed® (garlic extract, Allium sativum) and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBCs (0.02-0.04%) were obtained with three different sources of cinnamon oil. MBCs for three sources of oregano and lemongrass oils ranged from 0.14% to 0.30% and 0.10% to 0.65%, respectively, and for two thyme oils were 2.11% and 2.22%. The highest concentration (5%) of Allimed® tested resulted in MBCs to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBCs for all but one isolate. PMID:25685547

  10. An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

    PubMed Central

    Starliper, Clifford E.; Ketola, Henry G.; Noyes, Andrew D.; Schill, William B.; Henson, Fred G.; Chalupnicki, Marc A.; Dittman, Dawn E.

    2014-01-01

    Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments of captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine whether selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC’s) were determined for these four oils, Allimed® (garlic extract, Allium sativum) and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBCs (0.02–0.04%) were obtained with three different sources of cinnamon oil. MBCs for three sources of oregano and lemongrass oils ranged from 0.14% to 0.30% and 0.10% to 0.65%, respectively, and for two thyme oils were 2.11% and 2.22%. The highest concentration (5%) of Allimed® tested resulted in MBCs to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBCs for all but one isolate. PMID:25685547

  11. Zymogram patterns of Naegleria spp isolated from natural water sources in Taling Chan district, Bangkok.

    PubMed

    Tiewcharoen, Supathra; Komalamisra, Narumon; Junnu, Virach

    2004-06-01

    A genetic approach was cited for species detection of the ameba genus Naegleria using allozyme electrophoresis to characterize the trophozoite stage of three strains of Naegleria fowleri isolated from patients with primary amebic meningoencephalitis, five thermophilic (45 degrees C) Naegleria spp isolated from natural water sources in the Taling Chan district, and a reference control strain, Naegleria fowleri CDC VO 3081. Isoenzymes of ameba whole-cell extracts were analyzed by vertical polyacrylamide slab gel electrophoresis to determine whether there was any correlation between different strains of the ameba. The results showed that five out of fifteen enzymes; aldehyde oxidase (ALDOX), aldolase (ALD), a-glycerophosphate dehydrogenase (a-GPDH), xanthine dehydrogenase (XDH), and glutamate oxaloacetate transaminase (GOT), were undetectable in the pathogenic strains, while the other enzymes; esterase (EST), fumerase (FUM), glucose-6-phosphate dehydrogenase (G-6-PDH), glucose phosphate isomerase (GPI), isocitate dehydrogenase (IDH), lactate dehydrogenase (LDH), leucine aminopeptidase (LAP), malic enzyme (ME), glucose phosphomutase (GPM), and malate dehydrogenase (MDH), were detected. Naegleria fowleri strains were biochemically the most homogeneous. They showed intraspecific isoenzyme variation that allowed them to be grouped. In contrast, the allozyme patterns (EST 1-7, IDH) of Naegleria spp isolated from the environment showed interspecific isoenzyme variations from the pathogenic Naegleria strain. In conclusion, this study recognized the zymograms of the Naegleria fowleri strains were heterogenically different from the thermophilic 45 degrees C Naegleria spp isolated from the environment.

  12. Detection of Helicosporidium spp. in metagenomic DNA.

    PubMed

    Mancera, Norberto; Douma, Lauren G; James, Sheldon; Liu, Stephanie; Van, Amy; Boucias, Drion G; Tartar, Aurélien

    2012-09-15

    Distinct isolates of the invertebrate pathogenic alga Helicosporidium sp., collected from different insect hosts and different geographic locations, were processed to sequence the 18S rDNA and β-tubulin genes. The sequences were analyzed to assess genetic variation within the genus Helicosporidium and to design Helicosporidium-specific 18S rDNA primers. The specificity of these primers was demonstrated by testing not only on the Helicosporidium sp. isolates, but also on two trebouxiophyte algae known to be close Helicosporidium relatives, Prototheca wickerhamii and Prototheca zopfii. The genus-specific primers were used to develop a culture-independent assay aimed at detecting the presence of Helicosporidium spp. in environmental waters. The assay was based on the PCR amplification of 18SrDNA gene fragments from metagenomic DNA preparations, and it resulted in the amplification of detectable products for all sampled sites. Phylogenetic analyses that included the environmental sequences demonstrated that all amplification products clustered in a strongly supported, monophyletic Helicosporidium clade, thereby validating the metagenomic approach and the taxonomic origin of the produced environmental sequences. In addition, the phylogenetic analyses established that Helicosporidium spp. isolated from coleopteran hosts are more closely related to each other than they are to the isolate collected from a dipteran host. Finally, the phylogenetic trees depicted intergeneric relationships that supported a Helicosporidium-Prototheca cluster but did not support a Helicosporidium-Coccomyxa grouping, suggesting that pathogenicity to invertebrates evolved at least twice independently within the trebouxiophyte green algae. PMID:22609409

  13. Aeromonas spp.: An Emerging Nosocomial Pathogen

    PubMed Central

    Batra, Priyam; Mathur, Purva; Misra, Mahesh C

    2016-01-01

    Aeromonads are hallophillic, nonacid fast, nonspore forming, Gram-negative rods which are widely distributed in the soil, foodstuffs, and aquatic environment. Since times immemorial, they are important zoonotic pathogens of poikilotherms but are now emerging as important human pathogens. These emerging enteric pathogens flourish in the water distribution system by forming biofilms. They possess large number of virulence factors including inherent resistance to various antibiotics and ability to form biofilms using quorum sensing. These properties make them easy pathogens for human infections. Aeromonads are important enteric pathogens, but, with the growing level of immunosuppression in the population, they have been associated with various extraintestinal infections, such as skin and soft-tissue infections, traumatic wound infections, and lower respiratory tract/urinary tract infections. The average annual incidence of bacteremia in Southern Taiwan due to Aeromonas spp. was 76 cases/million inhabitants between 2008 and 2010. However, the incidence reported from Western countries is much lower. The case fatality rate among patients with Aeromonas bacteremia ranges from 27.5 to 46%. Aeromonads are universally resistant to the narrow-spectrum penicillin group of antibiotics such as penicillin, ampicillin, carbenicillin, and ticarcillin. They are however susceptible to piperacillin, azlocillin, second and third generation cephalosporins, and carbapenems. Most of the Aeromonas species are susceptible to aminoglycosides, tetracycline, chloramphenicol, trimethoprim-sulfamethoxazole, quinolones, and monobactams. This manuscript is a comprehensive systematic review of the literature available on Aeromonas spp. PMID:27013806

  14. Legionella spp. in Puerto Rico cooling towers

    SciTech Connect

    Negron-Alvira, A.; Perez-Suarez, I.; Hazen, T.C. )

    1988-10-01

    Water samples from air conditioning cooling towers receiving different treatment protocols on five large municipal buildings in San Juan, P.R., were assayed for various Legionella spp. and serogroups by using direct immunofluorescence. Several water quality parameters were also measured for each sample. Guinea pigs were inoculated with water samples to confirm pathogenicity and recover viable organisms. Legionella pneumophila serogroups 1 to 6, L. bozemanii, L. micdadei, L. dumoffii, and L. gormanii were observed in at least one of the cooling towers. L. pneumophila was the most abundant species; its density reached 10{sup 5} cells per ml, which is within the range that is considered potentially pathogenic to humans. A significantly higher density of L. pneumophila was observed in the cooling tower water that was not being treated with biocides. Percent respiration (INT) and total cell activity (acridine orange direct count) were inversely correlated with bacterial density. This study demonstrates that Legionella spp. are present in tropical air-conditioning cooling systems and that, without continuous biocide treatment, they may reach densities that present a health risk.

  15. Using Pseudomonas spp. for Integrated Biological Control.

    PubMed

    Stockwell, Virginia O; Stack, James P

    2007-02-01

    ABSTRACT Pseudomonas spp. have been studied for decades as model organisms for biological control of plant disease. Currently, there are three commercial formulations of pseudomonads registered with the U.S. Environmental Protection Agency for plant disease suppression, Bio-Save 10 LP, Bio-Save 11 LP, and BlightBan A506. Bio-Save 10 LP and Bio-Save 11 LP, products of Jet Harvest Solutions, Longwood, FL, contain Pseudomonas syringae strains ESC-10 and ESC-11, respectively. These products are applied in packinghouses to prevent postharvest fungal diseases during storage of citrus, pome, stone fruits, and potatoes. BlightBan A506, produced by NuFarm Americas, Burr Ridge, IL, contains P. fluorescens strain A506. BlightBan A506 is applied primarily to pear and apple trees during bloom to suppress the bacterial disease fire blight. Combining BlightBan A506 with the antibiotic streptomycin improves control of fire blight, even in areas with streptomycin-resistant populations of the pathogen. BlightBan A506 also may reduce fruit russet and mild frost injury. These biocontrol products consisting of Pseudomonas spp. provide moderate to excellent efficacy against multiple production constraints, are relatively easy to apply, and they can be integrated with conventional products for disease control. These characteristics will contribute to the adoption of these products by growers and packinghouses.

  16. Legionella spp. outdoors: colonization, communication and persistence.

    PubMed

    Hilbi, Hubert; Hoffmann, Christine; Harrison, Christopher F

    2011-06-01

    Bacteria of the genus Legionella persist in a wide range of environmental habitats, including biofilms, protozoa and nematodes. Legionellaceae are 'accidental' human pathogens that upon inhalation cause a severe pneumonia termed 'Legionnaires' disease'. The interactions of L. pneumophila with eukaryotic hosts are governed by the Icm/Dot type IV secretion system (T4SS) and more than 150 'effector proteins', which subvert signal transduction pathways and promote the formation of the replication-permissive 'Legionella-containing vacuole'. The Icm/Dot T4SS is essential to infect free-living protozoa, such as the amoeba Dictyostelium discoideum, as well as the nematode Caenorhabditis elegans, or mammalian macrophages. To adapt to different niches, L. pneumophila not only responds to exogenous cues, but also to endogenous signals, such as the α-hydroxyketone compound LAI-1 (Legionella autoinducer-1). The long-term adaptation of Legionella spp. is based on extensive horizontal DNA transfer. In fact, Legionella spp. have acquired canonical 'genomic islands' of prokaryotic origin, but also a number of eukaryotic genes. Since many aspects of Legionella virulence against environmental predators and immune phagocytes are similar, an understanding of Legionella ecology provides valuable insights into the pathogenesis of legionellaceae for humans. PMID:23761274

  17. Characterizing the nuclear proteome of Paracoccidioides spp.

    PubMed

    Oliveira, Lucas Nojosa; Casaletti, Luciana; Báo, Sônia Nair; Borges, Clayton Luiz; de Sousa Lima, Patrícia; de Almeida Soares, Célia Maria

    2016-10-01

    Paracoccidioidomycosis is an endemic disease in Latin America, caused by thermo dimorphic fungi of the genus Paracoccidioides. Although previous proteome analyses of Paracoccidioides spp. have been carried out, the nuclear subproteome of this pathogen has not been described. In this way, we aimed to characterize the nuclear proteome of Paracoccidioides species, in the yeast form. For that, yeast cells were disrupted and submitted to cell fractionation. The purity of the nuclear fraction was confirmed by fluorescence and electron microscopy. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) allowed the identification of 867 proteins. In order to support our enrichment method for nuclear proteins, bioinformatics analysis were applied that allowed the identification of 281 proteins with nuclear localization. The analysis revealed proteins related to DNA maintenance, gene expression, synthesis and processing of messenger and ribosomal RNAs, likewise proteins of nuclear-cytoplasmic traffic. It was also possible to detect some proteins that are poorly expressed, like transcription factors involved in important roles such as resistance to abiotic stress, sporulation, cellular growth and DNA and chromatin maintenance. This is the first descriptive nuclear proteome of Paracoccidioides spp. that can be useful as an important platform base for fungi-specific nuclear processes. PMID:27647238

  18. Using Pseudomonas spp. for Integrated Biological Control.

    PubMed

    Stockwell, Virginia O; Stack, James P

    2007-02-01

    ABSTRACT Pseudomonas spp. have been studied for decades as model organisms for biological control of plant disease. Currently, there are three commercial formulations of pseudomonads registered with the U.S. Environmental Protection Agency for plant disease suppression, Bio-Save 10 LP, Bio-Save 11 LP, and BlightBan A506. Bio-Save 10 LP and Bio-Save 11 LP, products of Jet Harvest Solutions, Longwood, FL, contain Pseudomonas syringae strains ESC-10 and ESC-11, respectively. These products are applied in packinghouses to prevent postharvest fungal diseases during storage of citrus, pome, stone fruits, and potatoes. BlightBan A506, produced by NuFarm Americas, Burr Ridge, IL, contains P. fluorescens strain A506. BlightBan A506 is applied primarily to pear and apple trees during bloom to suppress the bacterial disease fire blight. Combining BlightBan A506 with the antibiotic streptomycin improves control of fire blight, even in areas with streptomycin-resistant populations of the pathogen. BlightBan A506 also may reduce fruit russet and mild frost injury. These biocontrol products consisting of Pseudomonas spp. provide moderate to excellent efficacy against multiple production constraints, are relatively easy to apply, and they can be integrated with conventional products for disease control. These characteristics will contribute to the adoption of these products by growers and packinghouses. PMID:18944382

  19. Aeromonas spp.: An Emerging Nosocomial Pathogen.

    PubMed

    Batra, Priyam; Mathur, Purva; Misra, Mahesh C

    2016-01-01

    Aeromonads are hallophillic, nonacid fast, nonspore forming, Gram-negative rods which are widely distributed in the soil, foodstuffs, and aquatic environment. Since times immemorial, they are important zoonotic pathogens of poikilotherms but are now emerging as important human pathogens. These emerging enteric pathogens flourish in the water distribution system by forming biofilms. They possess large number of virulence factors including inherent resistance to various antibiotics and ability to form biofilms using quorum sensing. These properties make them easy pathogens for human infections. Aeromonads are important enteric pathogens, but, with the growing level of immunosuppression in the population, they have been associated with various extraintestinal infections, such as skin and soft-tissue infections, traumatic wound infections, and lower respiratory tract/urinary tract infections. The average annual incidence of bacteremia in Southern Taiwan due to Aeromonas spp. was 76 cases/million inhabitants between 2008 and 2010. However, the incidence reported from Western countries is much lower. The case fatality rate among patients with Aeromonas bacteremia ranges from 27.5 to 46%. Aeromonads are universally resistant to the narrow-spectrum penicillin group of antibiotics such as penicillin, ampicillin, carbenicillin, and ticarcillin. They are however susceptible to piperacillin, azlocillin, second and third generation cephalosporins, and carbapenems. Most of the Aeromonas species are susceptible to aminoglycosides, tetracycline, chloramphenicol, trimethoprim-sulfamethoxazole, quinolones, and monobactams. This manuscript is a comprehensive systematic review of the literature available on Aeromonas spp. PMID:27013806

  20. Lactobacillus spp. associated with early childhood caries.

    PubMed

    Svec, P; Sedlácek, I; Zácková, L; Nováková, D; Kukletová, M

    2009-01-01

    A group of 69 lactobacilli was isolated from caries lesions and root canals of early childhood caries (ECC) affected children treated in the Department of Pedodontics (Children's Teaching Hospital, Brno, Czech Republic). Biochemical and physiological properties of all strains were characterized by API 50 CH kit and conventional tube tests. The rep-PCR fingerprinting with the (GTG)(5) primer was used for genotypic grouping of the isolates. The (GTG)(5)-PCR fingerprinting grouped all analyzed strains into a few clusters in nearly full agreement with phenotype identification results and clarified the taxonomic position of 13 biochemically unidentified strains. In total, 20 strains of Lactobacillus fermentum, 17 L. rhamnosus, 14 L. casei/paracasei, 7 L. gasseri, 7 L. salivarius and 4 L. plantarum were identified. Mixtures of two or even three Lactobacillus spp. were isolated from a few root canal content samples. Results obtained by biotyping and (GTG)(5)-PCR were generally comparable except for L. gasseri strains that were not biochemically identified. The (GTG)(5)-PCR fingerprinting was shown to be quicker, easier to perform and more reliable than biotyping. Our results imply this molecular method as a good tool for screening and identification of Lactobacillus spp. inhabiting dental plaque.

  1. Aeromonas spp.: An Emerging Nosocomial Pathogen.

    PubMed

    Batra, Priyam; Mathur, Purva; Misra, Mahesh C

    2016-01-01

    Aeromonads are hallophillic, nonacid fast, nonspore forming, Gram-negative rods which are widely distributed in the soil, foodstuffs, and aquatic environment. Since times immemorial, they are important zoonotic pathogens of poikilotherms but are now emerging as important human pathogens. These emerging enteric pathogens flourish in the water distribution system by forming biofilms. They possess large number of virulence factors including inherent resistance to various antibiotics and ability to form biofilms using quorum sensing. These properties make them easy pathogens for human infections. Aeromonads are important enteric pathogens, but, with the growing level of immunosuppression in the population, they have been associated with various extraintestinal infections, such as skin and soft-tissue infections, traumatic wound infections, and lower respiratory tract/urinary tract infections. The average annual incidence of bacteremia in Southern Taiwan due to Aeromonas spp. was 76 cases/million inhabitants between 2008 and 2010. However, the incidence reported from Western countries is much lower. The case fatality rate among patients with Aeromonas bacteremia ranges from 27.5 to 46%. Aeromonads are universally resistant to the narrow-spectrum penicillin group of antibiotics such as penicillin, ampicillin, carbenicillin, and ticarcillin. They are however susceptible to piperacillin, azlocillin, second and third generation cephalosporins, and carbapenems. Most of the Aeromonas species are susceptible to aminoglycosides, tetracycline, chloramphenicol, trimethoprim-sulfamethoxazole, quinolones, and monobactams. This manuscript is a comprehensive systematic review of the literature available on Aeromonas spp.

  2. Characterizing the nuclear proteome of Paracoccidioides spp.

    PubMed

    Oliveira, Lucas Nojosa; Casaletti, Luciana; Báo, Sônia Nair; Borges, Clayton Luiz; de Sousa Lima, Patrícia; de Almeida Soares, Célia Maria

    2016-10-01

    Paracoccidioidomycosis is an endemic disease in Latin America, caused by thermo dimorphic fungi of the genus Paracoccidioides. Although previous proteome analyses of Paracoccidioides spp. have been carried out, the nuclear subproteome of this pathogen has not been described. In this way, we aimed to characterize the nuclear proteome of Paracoccidioides species, in the yeast form. For that, yeast cells were disrupted and submitted to cell fractionation. The purity of the nuclear fraction was confirmed by fluorescence and electron microscopy. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) allowed the identification of 867 proteins. In order to support our enrichment method for nuclear proteins, bioinformatics analysis were applied that allowed the identification of 281 proteins with nuclear localization. The analysis revealed proteins related to DNA maintenance, gene expression, synthesis and processing of messenger and ribosomal RNAs, likewise proteins of nuclear-cytoplasmic traffic. It was also possible to detect some proteins that are poorly expressed, like transcription factors involved in important roles such as resistance to abiotic stress, sporulation, cellular growth and DNA and chromatin maintenance. This is the first descriptive nuclear proteome of Paracoccidioides spp. that can be useful as an important platform base for fungi-specific nuclear processes.

  3. Legionella spp. in Puerto Rico cooling towers.

    PubMed Central

    Negrón-Alvíra, A; Pérez-Suarez, I; Hazen, T C

    1988-01-01

    Water samples from air conditioning cooling towers receiving different treatment protocols on five large municipal buildings in San Juan, P.R., were assayed for various Legionella spp. and serogroups by using direct immunofluorescence. Several water quality parameters were also measured for each sample. Guinea pigs were inoculated with water samples to confirm pathogenicity and recover viable organisms. Legionella pneumophila serogroups 1 to 6, L. bozemanii, L. micdadei, L. dumoffii, and L. gormanii were observed in at least one of the cooling towers. L. pneumophila was the most abundant species; its density reached 10(5) cells per ml, which is within the range that is considered potentially pathogenic to humans. A significantly higher density of L. pneumophila was observed in the cooling tower water that was not being treated with biocides. Percent respiration (INT) and total cell activity (acridine orange direct count) were inversely correlated with bacterial density. This study demonstrates that Legionella spp. are present in tropical air-conditioning cooling systems and that, without continuous biocide treatment, they may reach densities that present a health risk. PMID:3202625

  4. Distribution of pathogenic Naegleria spp in Thailand.

    PubMed

    Tiewcharoen, S; Junnu, V

    2001-01-01

    Research concerning the distribution, isolation, viability, ultrastructure, morphology and immunogenicity of Naegleria fowleri has been increasing in Thailand during 1988-2000. The distribution of the organism was carried out from 1985 to 1987 in Si Sa Ket and Ubon Rachathani Provinces, after the first fatal case was reported in Si Sa Ket. Since then in a 1998 survey of N. fowleri in stagnant water around industrial areas was carried out in Pathum Thani, Samut Prakan and Lopburi provinces. The results showed that 10% of pathogenic Naegleria belonged to species fowleri as characterized by morphology and the occurrence of pathogenesis in mice after nasal inoculation. In the same year, Nacapunchai et al (1999) determined the prevalence of amebae in aquatic habitat of human environments in five parts of Thailand during the summer. Fourteen percent of free living Naegleria spp were found in both soil and water resources. Recent studies of the ultrastructure, factors affecting the viability and SDS-PAGE electrophoretic patterns of 3 Thai strains of pathogenic Naegleria spp indicated their similarities in morphological characteristics of pathogenic reference control, Naegleria fowleri CDC VO 3081. Additional study using a genetic approach to species criteria using allozyme electrophoresis had been conducted.

  5. Antimicrobial activity and partial characterization of bacteriocin-like inhibitory substances produced by Lactobacillus spp. isolated from artisanal Mexican cheese.

    PubMed

    Heredia-Castro, Priscilia Y; Méndez-Romero, José I; Hernández-Mendoza, Adrián; Acedo-Félix, Evelia; González-Córdova, Aarón F; Vallejo-Cordoba, Belinda

    2015-12-01

    Lactobacillus spp. from Mexican Cocido cheese were shown to produce bacteriocin-like substances (BLS) active against Staphylococcus aureus,Listeria innocua,Escherichia coli, andSalmonella typhimurium by using the disk diffusion method. Crude extracts of Lactobacillus fermentum showed strong inhibitory activity against Staph. aureus, L. innocua, E. coli, and Salmonella cholerae. Complete inactivation of antimicrobial activity was observed after treatment of crude extracts with proteinase K, pronase, papain, trypsin, and lysozyme, confirming their proteinaceous nature. However, antimicrobial activity was partly lost for some of the crude extracts when treated with α-amylase, indicating that carbohydrate moieties were involved. The antimicrobial activity of the crude extracts was stable at 65°C for 30min over a wide pH range (2-8), and addition of potassium chloride, sodium citrate, ethanol, and butanol did not affect antibacterial activity. However, antimicrobial activity was lost after heating at 121°C for 15min, addition of methanol or Tween 80. Fourteen out of 18 Lactobacillus spp. showed antimicrobial activity against different test microorganisms, and 12 presented bacteriocin-like substances. Generation time and growth rate parameters indicated that the antimicrobial activity of crude extracts from 3 different strains was effective against the 4 indicator microorganisms. One of the crude extracts showed inhibition not only against gram-positive but also against gram-negative bacteria. Bacteriocin-like substances produced by this specific Lactobacillus strain showed potential for application as a food biopreservative. PMID:26476937

  6. Antimicrobial activity and partial characterization of bacteriocin-like inhibitory substances produced by Lactobacillus spp. isolated from artisanal Mexican cheese.

    PubMed

    Heredia-Castro, Priscilia Y; Méndez-Romero, José I; Hernández-Mendoza, Adrián; Acedo-Félix, Evelia; González-Córdova, Aarón F; Vallejo-Cordoba, Belinda

    2015-12-01

    Lactobacillus spp. from Mexican Cocido cheese were shown to produce bacteriocin-like substances (BLS) active against Staphylococcus aureus,Listeria innocua,Escherichia coli, andSalmonella typhimurium by using the disk diffusion method. Crude extracts of Lactobacillus fermentum showed strong inhibitory activity against Staph. aureus, L. innocua, E. coli, and Salmonella cholerae. Complete inactivation of antimicrobial activity was observed after treatment of crude extracts with proteinase K, pronase, papain, trypsin, and lysozyme, confirming their proteinaceous nature. However, antimicrobial activity was partly lost for some of the crude extracts when treated with α-amylase, indicating that carbohydrate moieties were involved. The antimicrobial activity of the crude extracts was stable at 65°C for 30min over a wide pH range (2-8), and addition of potassium chloride, sodium citrate, ethanol, and butanol did not affect antibacterial activity. However, antimicrobial activity was lost after heating at 121°C for 15min, addition of methanol or Tween 80. Fourteen out of 18 Lactobacillus spp. showed antimicrobial activity against different test microorganisms, and 12 presented bacteriocin-like substances. Generation time and growth rate parameters indicated that the antimicrobial activity of crude extracts from 3 different strains was effective against the 4 indicator microorganisms. One of the crude extracts showed inhibition not only against gram-positive but also against gram-negative bacteria. Bacteriocin-like substances produced by this specific Lactobacillus strain showed potential for application as a food biopreservative.

  7. INK128 Exhibits Synergy with Azoles against Exophiala spp. and Fusarium spp.

    PubMed Central

    Gao, Lujuan; Sun, Yi; He, Chengyan; Li, Ming; Zeng, Tongxiang; Lu, Qiaoyun

    2016-01-01

    Infections of Exophiala spp. and Fusarium spp. are often chronic and recalcitrant. Systemic disseminations, which mostly occur in immunocompromised patients, are often refractory to available antifungal therapies. The conserved target of rapamycin (TOR) orchestrates cell growth and proliferation in response to nutrients and growth factors, which are important for pathogenicity and virulence. INK128 is a second-generation ATP-competitive TOR inhibitor, which binds the TOR catalytic domain and selectively inhibits TOR. In the present study, we investigated the in vitro activities of INK128 alone and the interactions of INK128 with conventional antifungal drugs including itraconazole, voriconazole, posaconazole, and amphotericin B against 18 strains of Exophiala spp. and 10 strains of Fusarium spp. via broth microdilution checkerboard technique system adapted from Clinical and Laboratory Standards Institute broth microdilution method M38-A2. INK128 alone was inactive against all isolates tested. However, favorable synergistic effects between INK128 and voriconazole were observed in 61% Exophiala strains and 60% Fusarium strains, despite Fusarium strains exhibited high MIC values (4–8 μg/ml) against voriconazole. In addition, synergistic effects of INK128/itraconazole were shown in 33% Exophiala strains and 30% Fusarium strains, while synergy of INK128/posaconazole were observed in 28% Exophiala strains and 30% Fusarium strains. The effective working ranges of INK128 were 0.125–2 μg/ml and 1–4 μg/ml against Exophiala isolates and Fusarium isolates, respectively. No synergistic effect was observed when INK128 was combined with amphotericin B. No antagonism was observed in all combinations. In conclusion, INK128 could enhance the in vitro antifungal activity of voriconazole, itraconazole and posaconazole against Exophiala spp. and Fusarium spp., suggesting that azoles, especially voriconazole, combined with TOR kinase inhibitor might provide a potential strategy

  8. Exotic Small Mammals as Potential Reservoirs of Zoonotic Bartonella spp.

    PubMed Central

    Inoue, Kai; Kabeya, Hidenori; Hagiya, Keiko; Izumi, Yasuhito; Une, Yumi; Yoshikawa, Yasuhiro

    2009-01-01

    To evaluate the risk for emerging human infections caused by zoonotic Bartonella spp. from exotic small mammals, we investigated the prevalence of Bartonella spp. in 546 small mammals (28 species) that had been imported into Japan as pets from Asia, North America, Europe, and the Middle and Near East. We obtained 407 Bartonella isolates and characterized them by molecular phylogenetic analysis of the citrate synthase gene, gltA. The animals examined carried 4 zoonotic Bartonella spp. that cause human endocarditis and neuroretinitis and 6 novel Bartonella spp. at a high prevalence (26.0%, 142/546). We conclude that exotic small mammals potentially serve as reservoirs of several zoonotic Bartonella spp. PMID:19331727

  9. Rapid isolation of Yersinia spp. from feces.

    PubMed

    Weissfeld, A S; Sonnenwirth, A C

    1982-03-01

    Direct plating or cold enrichment or both have been used to isolate Yersinia spp. from feces. Freeze-shock double enrichment and KOH treatment have been recommended for recovery of Yersinia enterocolitica from surface waters and food, respectively. These techniques were evaluated as alternatives for rapid recovery of Yersinia spp. from feces. Stool samples were homogenized in buffered saline and autoclaved. Escherichia coli. Klebsiella pneumoniae, and Pseudomonas aeruginosa were each added to the suspension at a final concentration of 1.5 x 10(6) colony-forming units per ml. Yersinia cells were then added to a final concentration of 1.5 x 10(3), 1.5 x 10(4), 1.5 x 10(5), or 1.5 x 10(6) colony-forming units per ml. A total of 21 strains of Y. enterocolitica, 2 of Yersinia kristensenii, and 1 each of Yersinia intermedia and Yersinia fredriksenii were tested. For freeze-shock double enrichment, seeded stool samples were frozen overnight (-70 degrees C), transferred successively to m-tetrathionate broth (6 h. 37 degrees C) and selenite broth (2 h 37 degrees C), and plated on MacConkey, salmonella-shigella, and cellobiose-arginine-lysine agars for quantitation. For KOH treatment, seeded stool samples were mixed with 0.5% KOH at a ratio of 1:2 for 2 min and plated as described above. E. coli, K. pneumoniae, and P. aeruginosa were virtually eliminated after either method was used. All Yersinia strains were recovered after KOH treatment even at the lowest initial concentration (1.5 x 10(3) colony-forming units per ml). However, after freeze-shock double enrichment, not all strains were retrievable, and those isolates which were recovered were grown only from samples containing the highest number of Yersinia strains (1.5 x 10(6) colony-forming units per ml). KOH treatment of stool samples seems to be a viable substitute for more protracted methods of recovering Yersinia spp.

  10. The effect of co-administration of death camas (Zigadenus spp.) and low larkspur (Delphinium spp.) in cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In many rangeland settings, there is more than one potential poisonous plant. Two poisonous plants that are often found growing simultaneously in the same location are death camas (Zigadenus spp.) and low larkspur (Delphinium spp.). The objective of this study was to determine if co-administration...

  11. Contamination of public parks and squares from Guarulhos (São Paulo State, Brazil ) by Toxocara spp. and Ancylostoma spp.

    PubMed

    Marques, Jacó Pereira; Guimarães, Catarina de Rezende; Boas, Ailton Vilas; Carnaúba, Paulo Usignolo; Moraes, Josué de

    2012-01-01

    The contaminated soil with mammal feces is an important factor of risk to infection with zoonotic diseases. Amongst these zoonoses are visceral larva migrans and cutaneous larva migrans caused by Toxocara spp. and Ancylostoma spp., respectively. The aim of this study was to assess the environmental contamination by Toxocara spp. eggs and hookworms (Ancylostoma spp.) in public parks and squares in the city of Guarulhos, a metropolitan area of São Paulo, São Paulo State, Brazil. Soil samples were collected, between September and December 2010, and examined using the centrifugal flotation technique with sodium dichromate and zinc sulphate as well as the modified Baermann method. Notably, 35 (74.5%) of the 47 districts surveyed in Guarulhos possessed samples contaminated with Toxocara spp. and/or eggs or larvae of Ancylostoma spp. The frequency of Toxocara spp. and Ancylostoma spp. in the samples from public areas was 68.1% and 46.8%, respectively. Overall, the eastern side of Guarulhos is the region with the highest occurrence of causative agents of larva migrans. In all collection sites, the presence of feces from dogs and cats accompanied by their owners and stray animals were observed. Notably, it is important to adopt measures to control dog and cat breeding, to treat infected animals, and provide health education to the population.

  12. Persistent Giardia spp. and Trichuris spp. infection in maras (Dolichotis patagonum) at a zoo in Greece.

    PubMed

    Tahas, Stamatios Alan; Diakou, Anastasia

    2013-06-01

    The mara (Dolichotis patagonum) is a species classified as "Near Threatened" by the International Union for Conservation of Nature. In the wild, it inhabits only Argentina, but it is also kept in zoos around the world. In order to investigate the endoparasites of the maras kept in the Attica Zoological Park, Greece, four fecal examinations were performed in a period of 4 yr (2008-2011) by standard parasitologic methods. Cysts of the protozoan parasite Giardia spp. and eggs of the nematode Trichuris spp. were found in all four examinations. The possible routes of infection of the maras and the importance of these parasites to other animals and to humans are discussed. PMID:23805557

  13. Comparative genomics of Fructobacillus spp. and Leuconostoc spp. reveals niche-specific evolution of Fructobacillus spp.

    SciTech Connect

    Endo, Akihito; Tanizawa, Yasuhiro; Tanaka, Naoto; Maeno, Shintaro; Kumar, Himanshu; Shiwa, Yuh; Okada, Sanae; Yoshikawa, Hirofumi; Dicks, Leon; Nakagawa, Junichi; Arita, Masanori

    2015-12-29

    In this study, Fructobacillus spp. in fructose-rich niches belong to the family Leuconostocaceae. They were originally classified as Leuconostoc spp., but were later grouped into a novel genus, Fructobacillus , based on their phylogenetic position, morphology and specific biochemical characteristics. The unique characters, so called fructophilic characteristics, had not been reported in the group of lactic acid bacteria, suggesting unique evolution at the genome level. Here we studied four draft genome sequences of Fructobacillus spp. and compared their metabolic properties against those of Leuconostoc spp. As a result, Fructobacillus species possess significantly less protein coding sequences in their small genomes. The number of genes was significantly smaller in carbohydrate transport and metabolism. Several other metabolic pathways, including TCA cycle, ubiquinone and other terpenoid-quinone biosynthesis and phosphotransferase systems, were characterized as discriminative pathways between the two genera. The adhE gene for bifunctional acetaldehyde/alcohol dehydrogenase, and genes for subunits of the pyruvate dehydrogenase complex were absent in Fructobacillus spp. The two genera also show different levels of GC contents, which are mainly due to the different GC contents at the third codon position. In conclusion, the present genome characteristics in Fructobacillus spp. suggest reductive evolution that took place to adapt to specific niches.

  14. Proteasome Inhibitors Block Development of Plasmodium spp.

    PubMed Central

    Gantt, Soren M.; Myung, Joon Mo; Briones, Marcelo R. S.; Li, Wei Dong; Corey, E. J.; Omura, Satoshi; Nussenzweig, Victor; Sinnis, Photini

    1998-01-01

    Proteasomes degrade most of the proteins inside eukaryotic cells, including transcription factors and regulators of cell cycle progression. Here we show that nanomolar concentrations of lactacystin, a specific irreversible inhibitor of the 20S proteasome, inhibit development of the exoerythrocytic and erythrocytic stages of the malaria parasite. Although lactacystin-treated Plasmodium berghei sporozoites are still invasive, their development into exoerythrocytic forms (EEF) is inhibited in vitro and in vivo. Erythrocytic schizogony of P. falciparum in vitro is also profoundly inhibited when drug treatment of the synchronized parasites is prior, but not subsequent, to the initiation of DNA synthesis, suggesting that the inhibitory effect of lactacystin is cell cycle specific. Lactacystin reduces P. berghei parasitemia in rats, but the therapeutic index is very low. Along with other studies showing that lactacystin inhibits stage-specific transformation in Trypanosoma and Entamoeba spp., these findings highlight the potential of proteasome inhibitors as drugs for the treatment of diseases caused by protozoan parasites. PMID:9756786

  15. Helicobacter spp. other than H. pylori.

    PubMed

    Rossi, Mirko; Hänninen, Marja-Liisa

    2012-09-01

    Significant advances have been made over the last 12 months in the understanding of the biology of non-H. pylori Helicobacter species (NHPH). Several studies have investigated the association between NHPH and human disease, including Crohn's disease, lithiasis, liver disease, coronary disease, gastritis, and pyoderma gangrenosum-like ulcers. Novel Helicobacter taxa were identified in new vertebrate hosts, and new methodologies in the fields of identification of Helicobacter spp. and evaluation of antibiotic resistance were described. The genome of the first human-derived gastric NHPH strain (Helicobacter bizzozeronii CIII-1) was sequenced, and several studies elucidated functions of different genes in NHPH. A number of important investigations regarding pathogenesis and immunopathobiology of NHPH infections have been published including the description of a new urease in Helicobacter mustelae. Finally, the effects of the gut microbiota and probiotics on NHPH infections were investigated.

  16. [Antibiotic resistance analysis of Enterococcus spp. and Enterobacteriaceae spp. isolated from food].

    PubMed

    Korotkevich, Yu V

    2016-01-01

    The isolates from foods were screened for sensitivity to clinically significant antibiotics to assess the actual situation related to the prevalence of the antibiotic-resistant microorganisms in food. The goal of this work was to study the phenotypic characteristics of the antibiotic susceptibility of Enterobacteriaceae and Enterococcus spp. isolated from the good quality foods, and evaluation of the prevalence of tetracycline resistance in this groups of microbial contaminants. 68 strains of Enterobacteriaceae family and Enterococcus spp. isolated from poultry and livestock meat, pasteurized dairy products, acquired in the retail in the Moscow region, were studied. The disk-diffusion method (DDM) analysis showed a rather high prevalence of bacteria that are resistant and forming resistance to broad-spectrum antibiotics: in general 38% of Enterobacteriaceae strains and 40% of Enterococcus spp., isolated from meat products were resistant to tetracycline and doxycycline, and 21 and 33% - from dairy products, respectively; 26% of milk isolates and 54% of meat isolates were resistant to ampicillin. Considering that the tetracyclines is the most frequently used in animal husbandry and veterinary, the incidence and levels of tetracycline resistance were evaluated using tests with higher sensitivity to minimum inhibitory concentration (MIC), than the DDM. It was shown that among the Enterobacteriaceae strains 26% of isolates and 38% isolates were highly resistant to tetracycline (MIC ranged from 8 to 120 mg/kg) and 17-40% - among Enterococcus spp. These data obtained on a small number of samples, however, correspond to the frequency of tetracycline resistant strains detected in animal products in the EU (10-50%). Two multidrug-resistant enterobacteria strains - Klebsiella pneumoniae (farmer cheese) and Escherichia coli (minced turkey) were found among the .46 strains (4.4%), and they were resistant to 8 antibiotics.

  17. [Antibiotic resistance analysis of Enterococcus spp. and Enterobacteriaceae spp. isolated from food].

    PubMed

    Korotkevich, Yu V

    2016-01-01

    The isolates from foods were screened for sensitivity to clinically significant antibiotics to assess the actual situation related to the prevalence of the antibiotic-resistant microorganisms in food. The goal of this work was to study the phenotypic characteristics of the antibiotic susceptibility of Enterobacteriaceae and Enterococcus spp. isolated from the good quality foods, and evaluation of the prevalence of tetracycline resistance in this groups of microbial contaminants. 68 strains of Enterobacteriaceae family and Enterococcus spp. isolated from poultry and livestock meat, pasteurized dairy products, acquired in the retail in the Moscow region, were studied. The disk-diffusion method (DDM) analysis showed a rather high prevalence of bacteria that are resistant and forming resistance to broad-spectrum antibiotics: in general 38% of Enterobacteriaceae strains and 40% of Enterococcus spp., isolated from meat products were resistant to tetracycline and doxycycline, and 21 and 33% - from dairy products, respectively; 26% of milk isolates and 54% of meat isolates were resistant to ampicillin. Considering that the tetracyclines is the most frequently used in animal husbandry and veterinary, the incidence and levels of tetracycline resistance were evaluated using tests with higher sensitivity to minimum inhibitory concentration (MIC), than the DDM. It was shown that among the Enterobacteriaceae strains 26% of isolates and 38% isolates were highly resistant to tetracycline (MIC ranged from 8 to 120 mg/kg) and 17-40% - among Enterococcus spp. These data obtained on a small number of samples, however, correspond to the frequency of tetracycline resistant strains detected in animal products in the EU (10-50%). Two multidrug-resistant enterobacteria strains - Klebsiella pneumoniae (farmer cheese) and Escherichia coli (minced turkey) were found among the .46 strains (4.4%), and they were resistant to 8 antibiotics. PMID:27455596

  18. Morphological and Molecular Survey of Naegleria spp. in Water Bodies Used for Recreational Purposes in Rasht city, Northern Iran

    PubMed Central

    NIYYATI, Maryam; LASJERDI, Zohreh; ZAREIN-DOLAB, Saeed; NAZAR, Mahdieh; BEHNIAFAR, Hamed; MAHMOUDI, Mohammad Reza; NAZEMALHOSSEINI MOJARAD, Ehsan

    2015-01-01

    Background: Naegleria spp. is a free-living amoeba of which some species including N. fowleri and N. australeinsis are highly pathogenic in human and animals. These widespread amoebae could be found in different environmental sources particularly in aquatic resources of tropical and subtropical regions. The most important source of infection is via recreational water contact. Due to the lack of thorough research regarding species of Naegleria spp. in aquatic sources, the present study was conducted. Methods: In the present study, 60 samples were collected from recreational water resources of Rasht city, Guilan province, north of Iran. After filtering and culturing the samples, plates were examined by microscopic method and according to the page criteria. DNA of vahlkampfiid-positive samples were then extracted using phenol-chlorophorm method. Amoebae genus was identified by targeting the ITS-region and sequencing based-approaches. Results: Nine (15%) samples out of a 60 total samples were positive for Naegleria spp. of which seven belonged to potentially pathogenic N. australiensis. Two other strains were belonged to non-pathogenic N. pagei. Conclusion: The present research was the first report of occurrence of N. australiensis and N. pagei in Rasht city, north Iran. This study reflects the occurrence of Naegleria spp. in water sources of Guilan Province, Iran. PMID:26811717

  19. Antifungal activities of Hedychium essential oils and plant extracts against mycotoxigenic fungi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant-derived antifungal compounds are preferred to chemicals to reduce the risk of toxic effects on humans, livestock and the environment. Essential oil extracted from rhizomes and plant extracts of ornamental ginger lily (Hedychium spp.) were evaluated for their antifungal activity against two fu...

  20. Leptospira spp. in Rodents and Shrews in Germany

    PubMed Central

    Mayer-Scholl, Anne; Hammerl, Jens Andre; Schmidt, Sabrina; Ulrich, Rainer G.; Pfeffer, Martin; Woll, Dietlinde; Scholz, Holger C.; Thomas, Astrid; Nöckler, Karsten

    2014-01-01

    Leptospirosis is an acute, febrile disease occurring in humans and animals worldwide. Leptospira spp. are usually transmitted through direct or indirect contact with the urine of infected reservoir animals. Among wildlife species, rodents act as the most important reservoir for both human and animal infection. To gain a better understanding of the occurrence and distribution of pathogenic leptospires in rodent and shrew populations in Germany, kidney specimens of 2973 animals from 11 of the 16 federal states were examined by PCR. Rodent species captured included five murine species (family Muridae), six vole species (family Cricetidae) and six shrew species (family Soricidae). The most abundantly trapped animals were representatives of the rodent species Apodemus flavicollis, Clethrionomys glareolus and Microtus agrestis. Leptospiral DNA was amplified in 10% of all animals originating from eight of the 11 federal states. The highest carrier rate was found in Microtus spp. (13%), followed by Apodemus spp. (11%) and Clethrionomys spp. (6%). The most common Leptospira genomospecies determined by duplex PCR was L. kirschneri, followed by L. interrogans and L. borgpetersenii; all identified by single locus sequence typing (SLST). Representatives of the shrew species were also carriers of Leptospira spp. In 20% of Crocidura spp. and 6% of the Sorex spp. leptospiral DNA was detected. Here, only the pathogenic genomospecies L. kirschneri was identified. PMID:25062275

  1. PLS-Prediction and Confirmation of Hydrojuglone Glucoside as the Antitrypanosomal Constituent of Juglans Spp.

    PubMed

    Ellendorff, Therese; Brun, Reto; Kaiser, Marcel; Sendker, Jandirk; Schmidt, Thomas J

    2015-05-29

    Naphthoquinones (NQs) occur naturally in a large variety of plants. Several NQs are highly active against protozoans, amongst them the causative pathogens of neglected tropical diseases such as human African trypanosomiasis (sleeping sickness), Chagas disease and leishmaniasis. Prominent NQ-producing plants can be found among Juglans spp. (Juglandaceae) with juglone derivatives as known constituents. In this study, 36 highly variable extracts were prepared from different plant parts of J. regia, J. cinerea and J. nigra. For all extracts, antiprotozoal activity was determined against the protozoans Trypanosoma cruzi, T. brucei rhodesiense and Leishmania donovani. In addition, an LC-MS fingerprint was recorded for each extract. With each extract's fingerprint and the data on in vitro growth inhibitory activity against T. brucei rhodesiense a Partial Least Squares (PLS) regression model was calculated in order to obtain an indication of compounds responsible for the differences in bioactivity between the 36 extracts. By means of PLS, hydrojuglone glucoside was predicted as an active compound against T. brucei and consequently isolated and tested in vitro. In fact, the pure compound showed activity against T. brucei at a significantly lower cytotoxicity towards mammalian cells than established antiprotozoal NQs such as lapachol.

  2. Detection of Sarcocystis spp. infection in bobcats (Lynx rufus).

    PubMed

    Verma, S K; Calero-Bernal, R; Lovallo, M J; Sweeny, A R; Grigg, M E; Dubey, J P

    2015-09-15

    The protozoan Sarcocystis neurona is an important cause of severe clinical disease of horses (called equine protozoal myeloencephalitis, EPM), marine mammals, companion animals, and several species of wildlife animals in the Americas. The Virginia opossum (Didelphis virginiana) is its definitive host in the USA and other animals act as intermediate or aberrant hosts. Samples of tongue and heart from 35 bobcats hunted for fur and food from Mississippi State, USA in February, 2014 were used for the present study. Muscles were examined for Sarcocystis infection by microscopic examination of either unfixed muscle squash preparations or pepsin digests, by histopathology of fixed samples, and by molecular methods. Sarcocystis-like bradyzoites were found in digests of 14 hearts and 10 tongues of 35 bobcats. In histological sections, sarcocysts were found in 26 of 35 bobcats; all appeared relatively thin-walled similar to S. felis sarcocysts under light microscope at 1000× magnification. S. neurona-like sarcocysts having thickened villar tips were seen in unstained muscle squash of tongue of two bobcats and PCR-DNA sequencing identified them definitively as S. neurona-like parasites. DNA extracted from bradyzoites obtained from tongue and heart muscle digests was analyzed by PCR-DNA sequencing at the ITS1 locus. Results indicated the presence of S. neurona-like parasite in 26 of 35 samples. ITS1 sequences identical to S. dasypi were identified in 3 bobcats, 2 of which were also co-infected with S. neurona-like parasite. The high prevalence of sarcocysts in bobcat tissues suggested an efficient sylvatic cycle of Sarcocystis spp. in the remote regions of Mississippi State with the bobcat as a relevant intermediate host.

  3. Detection of Sarcocystis spp. infection in bobcats (Lynx rufus).

    PubMed

    Verma, S K; Calero-Bernal, R; Lovallo, M J; Sweeny, A R; Grigg, M E; Dubey, J P

    2015-09-15

    The protozoan Sarcocystis neurona is an important cause of severe clinical disease of horses (called equine protozoal myeloencephalitis, EPM), marine mammals, companion animals, and several species of wildlife animals in the Americas. The Virginia opossum (Didelphis virginiana) is its definitive host in the USA and other animals act as intermediate or aberrant hosts. Samples of tongue and heart from 35 bobcats hunted for fur and food from Mississippi State, USA in February, 2014 were used for the present study. Muscles were examined for Sarcocystis infection by microscopic examination of either unfixed muscle squash preparations or pepsin digests, by histopathology of fixed samples, and by molecular methods. Sarcocystis-like bradyzoites were found in digests of 14 hearts and 10 tongues of 35 bobcats. In histological sections, sarcocysts were found in 26 of 35 bobcats; all appeared relatively thin-walled similar to S. felis sarcocysts under light microscope at 1000× magnification. S. neurona-like sarcocysts having thickened villar tips were seen in unstained muscle squash of tongue of two bobcats and PCR-DNA sequencing identified them definitively as S. neurona-like parasites. DNA extracted from bradyzoites obtained from tongue and heart muscle digests was analyzed by PCR-DNA sequencing at the ITS1 locus. Results indicated the presence of S. neurona-like parasite in 26 of 35 samples. ITS1 sequences identical to S. dasypi were identified in 3 bobcats, 2 of which were also co-infected with S. neurona-like parasite. The high prevalence of sarcocysts in bobcat tissues suggested an efficient sylvatic cycle of Sarcocystis spp. in the remote regions of Mississippi State with the bobcat as a relevant intermediate host. PMID:26138150

  4. Ilarviruses of Prunus spp.: a continued concern for fruit trees.

    PubMed

    Pallas, V; Aparicio, F; Herranz, M C; Amari, K; Sanchez-Pina, M A; Myrta, A; Sanchez-Navarro, J A

    2012-12-01

    Prunus spp. are affected by a large number of viruses, causing significant economic losses through either direct or indirect damage, which results in reduced yield and fruit quality. Among these viruses, members of the genus Ilarvirus (isometric labile ringspot viruses) occupy a significant position due to their distribution worldwide. Although symptoms caused by these types of viruses were reported early in the last century, their molecular characterization was not achieved until the 1990s, much later than for other agronomically relevant viruses. This was mainly due to the characteristic liability of virus particles in tissue extracts. In addition, ilarviruses, together with Alfalfa mosaic virus, are unique among plant viruses in that they require a few molecules of the coat protein in the inoculum in order to be infectious, a phenomenon known as genome activation. Another factor that has made the study of this group of viruses difficult is that infectious clones have been obtained only for the type member of the genus, Tobacco streak virus. Four ilarviruses, Prunus necrotic ringspot virus, Prune dwarf virus, Apple mosaic virus, and American plum line pattern virus, are pathogens of the main cultivated fruit trees. As stated in the 9th Report of the International Committee on Taxonomy of Viruses, virions of this genus are "unpromising subjects for the raising of good antisera." With the advent of molecular approaches for their detection and characterization, it has been possible to get a more precise view of their prevalence and genome organization. This review updates our knowledge on the incidence, genome organization and expression, genetic diversity, modes of transmission, and diagnosis, as well as control of this peculiar group of viruses affecting fruit trees.

  5. Detection of Sarcocystis spp. infection in bobcats (Lynx rufus)

    PubMed Central

    Verma, S. K.; Calero-Bernal, R.; Lovallo, M. J.; Sweeny, A. R.; Grigg, M. E.; Dubey, J. P.

    2015-01-01

    The protozoan Sarcocystis neurona is an important cause of severe clinical disease of horses (called equine protozoal myeloencephalitis, EPM), marine mammals, companion animals, and several species of wildlife animals in the Americas. The Virginia opossum (Didelphis virginiana) is its definitive host in the USA and other animals act as intermediate or aberrant hosts. Samples of tongue and heart from 35 bobcats hunted for fur and food from Mississippi State, USA in February, 2014 were used for the present study. Muscles were examined for Sarcocystis infection by microscopic examination of either unfixed muscle squash preparations or pepsin digests, by histopathology of fixed samples, and by molecular methods. Sarcocystis-like bradyzoites were found in digests of 14 hearts and 10 tongues of 35 bobcats. In histological sections, sarcocysts were found in 26 of 35 bobcats; all appeared relatively thin-walled similar to S. felis sarcocysts under light microscope at 1000x magnification. S. neurona-like sarcocysts having thickened villar tips were seen in unstained muscle squash of tongue of two bobcats and PCR-DNA sequencing identified them definitively as S. neurona-like parasite. DNA extracted from bradyzoites obtained from tongue and heart muscle digests was analyzed by PCR-DNA sequencing at the ITS1 locus. Results indicated the presence of S. neurona-like parasite in 26 of 35 samples. ITS1 sequences identical to S. dayspi were identified in 3 bobcats, 2 of which were also co-infected with S. neurona-like parasite. The high prevalence of sarcocysts in bobcat tissues suggested an efficient sylvatic cycle of Sarcocystis spp. in the remote regions of Mississippi State with the bobcat as a relevant intermediate host. PMID:26138150

  6. Extraction process

    SciTech Connect

    Dente, M.; Porcari, G.; Robinson, L. F.

    1985-08-06

    Hydrocarbon liquids are recovered from oil shale and other solids containing organic matter by passing a liquid organic solvent downwardly through an extraction zone in contact with said solids at an elevated pressure sufficient to maintain said solvent in the liquid phase and at a temperature below about 900/sup 0/ F., preferably between about 650/sup 0/ F. and about 900/sup 0/ F., in order to extract hydrocarbons from the solids into the solvent. The extracted hydrocarbons are then recovered from the solvent by fractionation. Normally, heat is supplied to the extraction zone by passing a hot, nonoxidizing gas, preferably an oxygen-free gas generated within the process, downwardly through the extraction zone in cocurrent flow with the liquid organic solvent.

  7. Giardia and Cryptosporidium spp. dissemination during wastewater treatment and comparative detection via immunofluorescence assay (IFA), nested polymerase chain reaction (nested PCR) and loop mediated isothermal amplification (LAMP).

    PubMed

    Gallas-Lindemann, Carmen; Sotiriadou, Isaia; Plutzer, Judit; Noack, Michael J; Mahmoudi, Mohammad Reza; Karanis, Panagiotis

    2016-06-01

    Environmental water samples from the Lower Rhine area in Germany were investigated via immunofluorescence assays (IFAs), nested polymerase chain reaction (nested PCR) and loop-mediated isothermal amplification (LAMP) to detect the presence of Giardia spp. (n=185) and Cryptosporidium spp. (n=227). The samples were concentrated through filtration or flocculation, and oocysts were purified via centrifugation through a sucrose density gradient. For all samples, IFA was performed first, followed by DNA extraction for the nested PCR and LAMP assays. Giardia cysts were detected in 105 samples (56.8%) by IFA, 62 samples (33.5%) by nested PCR and 79 samples (42.7%) by LAMP. Cryptosporidium spp. were detected in 69 samples (30.4%) by IFA, 95 samples (41.9%) by nested PCR and 99 samples (43.6%) by LAMP. According to these results, the three detection methods are complementary for monitoring Giardia and Cryptosporidium in environmental waters.

  8. An analytical approach based on ESI-MS, LC-MS and PCA for the quali-quantitative analysis of cycloartane derivatives in Astragalus spp.

    PubMed

    Napolitano, Assunta; Akay, Seref; Mari, Angela; Bedir, Erdal; Pizza, Cosimo; Piacente, Sonia

    2013-11-01

    Astragalus species are widely used as health foods and dietary supplements, as well as drugs in traditional medicine. To rapidly evaluate metabolite similarities and differences among the EtOH extracts of the roots of eight commercial Astragalus spp., an approach based on direct analyses by ESI-MS followed by PCA of ESI-MS data, was carried out. Successively, quali-quantitative analyses of cycloartane derivatives in the eight Astragalus spp. by LC-ESI-MS(n) and PCA of LC-ESI-MS data were performed. This approach allowed to promptly highlighting metabolite similarities and differences among the various Astragalus spp. PCA results from LC-ESI-MS data of Astragalus samples were in reasonable agreement with both PCA results of ESI-MS data and quantitative results. This study affords an analytical method for the quali-quantitative determination of cycloartane derivatives in herbal preparations used as health and food supplements.

  9. Relative Frequency, Characteristics, and Antimicrobial Susceptibility Patterns of Vibrio spp., Aeromonas spp., Chromobacterium violaceum, and Shewanella spp. in the Northern Territory of Australia, 2000–2013

    PubMed Central

    McAuliffe, Gary N.; Hennessy, Jann; Baird, Robert W.

    2015-01-01

    Vibrio, Aeromonas, Chromobacterium violaceum, and Shewanella (VACS) are water-associated Gram-negative organisms that can cause a variety of infections. The frequency, patient characteristics, and antimicrobial susceptibilities for 468 isolates from 442 patients from the Northern Territory were reviewed. Aeromonas spp. (312 of 468; 67%) were most commonly isolated followed by Vibrio spp. (71 of 468; 15%), Shewanella spp. (61 of 468; 13%), and C. violaceum (24 of 468; 5%). A strong male predominance was found (male to female ratio of 2.3:1). Skin and soft tissue isolations (373 of 468; 80%) from lower limb infections (222 of 371; 60%) were the most common clinical manifestation. The episodes were usually polymicrobial (281 of 468; 60%). Coisolates included Staphylococcus aureus (137 of 468; 29%), β-hemolytic streptococci (74 of 468; 16%), enterobacteriaceae (111 of 468; 24%), non-fermentative Gram-negative bacilli (35 of 468; 7%), and other VACS organisms (37 of 468; 8%). Antimicrobial resistance of VACS organisms to ciprofloxacin (0–4%), cefepime (0–3%), and gentamicin (0–0.8%) and Vibrio spp., Aeromonas spp., and Shewanella to cotrimoxazole (0–3%) was rarely shown. For water-associated lower limb skin and soft tissue infections in the tropics, clinicians should consider empirical antimicrobial therapy with agents active against S. aureus and VACS organisms. PMID:25548380

  10. Elucidation of Leucaena leucocephala anthelmintic-like phytochemicals and the ultrastructural damage generated to eggs of Cooperia spp.

    PubMed

    von Son-de Fernex, Elke; Alonso-Díaz, Miguel Ángel; Mendoza-de Gives, Pedro; Valles-de la Mora, Braulio; González-Cortazar, Manases; Zamilpa, Alejandro; Castillo Gallegos, Epigmenio

    2015-11-30

    Leucaena leucocephala is a tropical forage legume suggested as an alternative method to control gastrointestinal parasitism in ruminants. This study: (1) performed a bio-guided fractionation of an aqueous extract of L. leucocephala using the egg hatch assay (EHA) to identify the anthelmintic (AH)-like phytochemicals present in fresh leaves, and (2) assessed the ultrastructural damage to eggs of Cooperia spp. after incubation with the final fraction. Phytochemicals were isolated using silica gel columns and identified using high performance liquid chromatography and standards for comparison. The final fraction was evaluated using EHA at 0.06, 0.125, 0.250, 0.500 and 1.1 mg ml(-1). The lethal concentration to inhibit 50% of Cooperia spp. egg hatching (LC50) was calculated using a Probit analysis. Scanning and transmission electron microscopy revealed the ultrastructural changes present in Cooperia spp. eggs. Bio-guided isolation procedures led to the recognition of an active fraction (LlC1F3) mainly composed of quercetin (82.21%) and caffeic acid (13.42%) which inhibited 90.49 ± 2.8% of Cooperia spp. egg hatching (P<0.05), and an LC50 of 0.06 ± 0.14 mg ml(-1). Scanning electron microscopy (SEM) showed eggs exposed to the active fraction had an irregular external layer with small projections and ruptures of lateral eggshell walls. Transmission electron microscopy (TEM) showed changes to Cooperia spp. eggs in electro-density, including the thickness of the eggshell layers and fractures after incubation with the final fraction (LlC1F3). Changes in bioactivity after purification suggest synergistic interactions between quercetin and caffeic acid. PMID:26477279

  11. Elucidation of Leucaena leucocephala anthelmintic-like phytochemicals and the ultrastructural damage generated to eggs of Cooperia spp.

    PubMed

    von Son-de Fernex, Elke; Alonso-Díaz, Miguel Ángel; Mendoza-de Gives, Pedro; Valles-de la Mora, Braulio; González-Cortazar, Manases; Zamilpa, Alejandro; Castillo Gallegos, Epigmenio

    2015-11-30

    Leucaena leucocephala is a tropical forage legume suggested as an alternative method to control gastrointestinal parasitism in ruminants. This study: (1) performed a bio-guided fractionation of an aqueous extract of L. leucocephala using the egg hatch assay (EHA) to identify the anthelmintic (AH)-like phytochemicals present in fresh leaves, and (2) assessed the ultrastructural damage to eggs of Cooperia spp. after incubation with the final fraction. Phytochemicals were isolated using silica gel columns and identified using high performance liquid chromatography and standards for comparison. The final fraction was evaluated using EHA at 0.06, 0.125, 0.250, 0.500 and 1.1 mg ml(-1). The lethal concentration to inhibit 50% of Cooperia spp. egg hatching (LC50) was calculated using a Probit analysis. Scanning and transmission electron microscopy revealed the ultrastructural changes present in Cooperia spp. eggs. Bio-guided isolation procedures led to the recognition of an active fraction (LlC1F3) mainly composed of quercetin (82.21%) and caffeic acid (13.42%) which inhibited 90.49 ± 2.8% of Cooperia spp. egg hatching (P<0.05), and an LC50 of 0.06 ± 0.14 mg ml(-1). Scanning electron microscopy (SEM) showed eggs exposed to the active fraction had an irregular external layer with small projections and ruptures of lateral eggshell walls. Transmission electron microscopy (TEM) showed changes to Cooperia spp. eggs in electro-density, including the thickness of the eggshell layers and fractures after incubation with the final fraction (LlC1F3). Changes in bioactivity after purification suggest synergistic interactions between quercetin and caffeic acid.

  12. Pseudo-nitzschia spp. (Bacillariophyceae) and dissolved organic matter (DOM) dynamics in the Ebro Delta (Alfacs Bay, NW Mediterranean Sea)

    NASA Astrophysics Data System (ADS)

    Loureiro, Sofia; Garcés, Esther; Fernández-Tejedor, Margarita; Vaqué, Dolors; Camp, Jordi

    2009-08-01

    Samples were collected during one annual cycle (April 2007-March 2008) at Alfacs Bay (NW Mediterranean Sea) central station in order to assess the influence of organic nutrients in the growth of the microalgae assemblage, with special reference to Pseudo-nitzschia spp. This potentially toxic diatom forms natural and recurrent blooms in the study area. To assess further the relationship between Pseudo-nitzschia spp. and nutrients an enrichment experiment with high molecular weight dissolved organic matter (HMWDOM) was performed with field samples obtained during a Pseudo-nitzschia spp. bloom. HMWDOM was extracted from water collected at Alfacs Bay. Five bioassays were prepared: N + P (seawater with addition of nitrate and phosphate), DOM (addition of HMWDOM), (-N + P) + DOM (nitrogen deficient, with addition of phosphate and HMWDOM), (N + P) + DOM (addition of nitrate, phosphate and HMWDOM), seawater control (without added nutrients), and B + DOM (control of bacteria, without microalgae). The experiment was run in batch mode over 4 days. Results from the field study revealed that the concentrations of organic nutrients mostly surpassed the inorganic pool. Pseudo-nitzschia spp. was the most frequent and abundant taxa of the microalgae community. The micro-planktonic assemblage was arranged according to a seasonal factor (ANOSIM and cluster analysis). DON, nitrate and silicate were the most important abiotic parameters contributing to the dissimilarities between seasons (SIMPER analysis) and thereby potentially influencing the seasonal distribution of microalgae in the representative station. In the experimental investigation, Pseudo-nitzschia cells increased by the end of the experiment in the DOM bioassay but no respective increase was observed for chlorophyll a. This could point to an acquisition of nutrients through the DOM fraction that would conjointly reduce the need of chlorophyll a. The data obtained suggest that organic nutrients may exert an important role

  13. Prevalence of Salmonella spp. in pet turtles and their environment

    PubMed Central

    Back, Du-San; Shin, Gee-Wook; Wendt, Mitchell

    2016-01-01

    Pet turtles are known as a source of Salmonella infection to humans when handled in captivity. Thirty four turtles purchased from pet shops and online markets in Korea were examined to determine whether the turtles and their environment were contaminated with Salmonella spp. Salmonella spp. were isolated from fecal samples of 17 turtles. These isolates were identified as S. enterica through 16S rRNA gene sequencing. The isolation rate of Salmonella spp. from the soil and water samples increased over time. We concluded that a high percentage of turtles being sold in pet shops were infected with Salmonella spp., and their environments tend to become contaminated over time unless they are maintained properly. These results indicate that pet turtles could be a potential risk of salmonellosis in Korea. PMID:27729933

  14. Incidence and inactivation of Listeria spp. on frozen shrimp

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Foodborne illness outbreaks occasionally occur as a result of microbiologically contaminated crustaceans, including shrimp. Foodborne pathogens occasionally found on shrimp include Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and Vibrios. In this study the microbiological qualit...

  15. An alternative bacteriological medium for the isolation of Aeromonas spp.

    USGS Publications Warehouse

    Jenkins, J.A.; Taylor, P.W.

    1995-01-01

    Two solid bacteriologic media were compared for cultivating Aeromonas spp. from piscine sources: the Rimler-Shotts (RS) medium and a starch-glutamate-ampicillin-penicillin-based medium (SGAP-10C) used for the recovery of Aeromonas spp. from water samples. The selective and differential capacities of the media were assessed March through October 1992 by recovery rate and phenotype of 99 isolates representing 15 genera of bacteria. Recovery frequency of Aeromonas spp. (n = 62) was similar at 97% on RS and 95% on SGAP-10C. The SGAP-10C medium proved to be more specific than RS toward Aeromonas species (P ≤ 0.005). Use of SGAP-10C at 24 C for 48 hr offers a better choice for the laboratory recovery of Aeromonas spp. from clinical fish specimens.

  16. Physiology and immunology of mucosal barriers in catfish (Ictalurus spp.)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The mucosal barriers of catfish (Ictalurus spp.) constitute the first line of defense against pathogen invasion while simultaneously carrying out a diverse array of other critical physiological processes, including nutrient adsorption, osmoregulation, waste excretion, and environmental sensing. Catf...

  17. Biofouling of groundwater distribution systems by Thiothrix spp.

    SciTech Connect

    Brigmon, R.L.; Martin, H.W.; Aldrich, H.C.

    1995-12-01

    Thiothrix spp., sulfide oxidizing filamentous bacteria, were found to be the main bacterial component of aquatic biofilms causing biofouling in selected municipal water storage tanks, private wells, and drip irrigation systems in Florida. The water originated from the upper Floridan aquifer and associated aquifers in Central and North Florida. Samples were examined where visible biofilms had a white, slimy, filamentous appearance indicative of Thiothrix spp. The detection of Thiothrix spp. was confirmed by enzyme-liked immunosorbent assay (ELISA). These observations confirm that these bacteria and associated extracellular material play an important role in formation of biofilms, which in turn may induce physical changes leading to significant biofouling. These studies suggest that Thiothrix spp.-associated biofouling occurs at an interface where reduced sulfide-containing water contacts aerated water and a surface or substrate is available for attachment.

  18. Symbiotic relationship of Thiothrix spp. with an echinoderm

    SciTech Connect

    Brigmon, R.L.; De Ridder, C.

    1998-09-01

    Thiothrix-like bacteria have been reported as symbionts in invertebrates from sulfide-rich habitats. Isolation of these symbiotic Thiothrix-like bacteria has failed, and the organisms have not been previously identified with certainty. The genus Thiothrix was created for ensheathed filamentous bacteria that oxidize sulfide and deposit sulfur granules internally, attach to substrates, produce gliding gonidia, and form rosettes. Immunoassay procedures were used to investigate the symbiotic relationship of Thiothrix spp. in the intestinal cecum of the spatangoid species Echinocardium cordatum. Thiothrix spp. were identified in nodule samples from E. cordatum digestive tubes based on microscopic examination, enzyme-linked immunosorbent assay, and indirect immunofluorescence. Thiothrix spp. protein made up as much as 84% of the total protein content of the nodules. This is the first identification of Thiothrix spp. internally symbiotic with marine invertebrates.

  19. [Echocardiopgraphy in European tortoises (Testudo spp.)].

    PubMed

    Prütz, Maike; Fehr, Michael; Mathes, Karina; Hungerbühler, Stephan

    2016-01-01

    An echocardiographic examination was carried out in 71 European tortoises (Testudo spp.) via the cervical-brachial acoustic windows. Simultaneously an electrocardiographic examination was performed. The inflow- and outflow tract of the heart were presented in frontal and sagittal longitudinal sections in B-mode. Within B-mode the size (diameter and area) of the atria and the ventricle (Cavum dorsale), the ventricular wall thickness and the diameter of the origin of the right aorta and of the right Arteria pulmonalis were measured. Also, the fractional shortening (FS%) and a fractional area shortening (FAS%) were calculated for the Cavum dorsale. Standard values for these cardiac parameters were determined for four different tortoise groups (depending on their carapace lengths). The direction of blood flow within the heart could be assessed via colour flow Doppler. By using pulsed-wave Doppler examinations of the inflow- and outflow tract the velocities, pressure gradients, velocity-time-integrals and acceleration- and deceleration times could be determined from the recorded inflow and outflow patterns and standard values were established for these parameters as well. PMID:27169156

  20. Recovery of Arcobacter spp. from nonlivestock species.

    PubMed

    Wesley, Irene V; Schroeder-Tucker, Linda

    2011-09-01

    The genus Arcobacter encompasses campylobacter-like organisms that grow in air at 25 degrees C. Arcobacter has been detected or isolated from clinically healthy livestock as well as aborted fetuses and has been presumptively identified as either Campylobacter or Leptospira, based on its growth in selective semisolid media. Because reports from nonlivestock species are limited, this study examined nine presumptive isolates of Arcobacter spp. from an alpaca (Vicugna pacos), black rhinoceros (Diceros bicornis), white rhinoceros (Ceratotherium simum), gorilla (Troglodytes gorilla), gazelle (Eudorcas thomsoni), rhea (Rhea americana), and aborted equine fetuses. Seven of these nine phenotypically identified isolates of Arcobacter were confirmed by a multiplex polymerase chain reaction assay. The remaining two isolates were subsequently identified as Arcobacter skirrowii (Case 5) and Campylobacter jejuni (Case 6) by sequence analysis of a 527-base pair fragment of the 16S rRNA gene. Together, these cases underscore the challenges to a clinical laboratory of identifying Arcobacter in cases which mimic vibrionic abortion or leptospirosis. PMID:22950328

  1. Current Knowledge of Trichosporon spp. and Trichosporonosis

    PubMed Central

    Colombo, Arnaldo L.; Padovan, Ana Carolina B.; Chaves, Guilherme M.

    2011-01-01

    Summary: Trichosporon spp. are basidiomycetous yeast-like fungi found widely in nature. Clinical isolates are generally related to superficial infections. However, this fungus has been recognized as an opportunistic agent of invasive infections, mostly in cancer patients and those exposed to invasive medical procedures. It is possible that the ability of Trichosporon strains to form biofilms on implanted devices, the presence of glucuronoxylomannan in their cell walls, and the ability to produce proteases and lipases are all factors likely related to the virulence of this genus and therefore may account for the progress of invasive trichosporonosis. Disseminated trichosporonosis has been increasingly reported worldwide and represents a challenge for both diagnosis and species identification. Phenotypic identification methods are useful for Trichosporon sp. screening, but only molecular methods, such as IGS region sequencing, allow the complete identification of Trichosporon isolates at the species level. Methods for the diagnosis of invasive trichosporonosis include PCR-based methods, Luminex xMAP technology, and, more recently, proteomics. Treating patients with trichosporonosis remains a challenge because of limited data on the in vitro and in vivo activities of antifungal drugs against clinically relevant species of the genus. Despite the mentioned limitations, the use of antifungal regimens containing triazoles appears to be the best therapeutic approach. PMID:21976604

  2. [Isolation of Candida spp. from ascites in cirrhotic patients].

    PubMed

    Saludes, Paula; Araguás, Cristina; Sánchez-Delgado, Jordi; Dalmau, Blai; Font, Bernat

    2016-10-01

    The isolation of Candida spp. in ascites of cirrhotic patients is an uncommon situation in clinical practice. Factors that have been associated with increased susceptibility to primary fungal peritonitis are exposure to broad-spectrum antibiotics and immunosuppression, a typical situation of these patients. We report seven episodes of Candida spp. isolation in ascites of cirrhotic patients detected in our hospital during the past 15years.

  3. Cefotaxime resistance and outcome of Klebsiella spp bloodstream infection.

    PubMed

    Ortega, M; Marco, F; Soriano, A; Almela, M; Martínez, J A; López, J; Pitart, C; Mensa, J

    2011-12-01

    We attempt to describe the epidemiology and outcome associated with cefotaxime-resistant (CTX-R) Klebsiella spp bacteraemia. Klebsiella spp bloodstream infection episodes prospectively collected through a blood culture surveillance programme from January 1991 to December 2008 in a single institution were analysed. A total of 910 monomicrobial episodes of Klebsiella spp bacteraemia were identified during the study period. The most important sources were from urinary tract infection, unknown sources, billiary focus and catheter related infection. There were 112 (12%) CTX-R isolates. Out of 112 isolates, 98 were CTX-R by Extended-Spectrum β-Lactamase production. Shock on presentation and mortality were significantly more frequent in CTX-R than in CTX susceptible isolates. Inappropriate empirical therapy was received in 50 (45%) cases in the CTX-R Klebsiella spp group (13 cases of death, 26%). Predictive factors associated with CTX-R Klebsiella spp isolate were: previous β-lactam therapy (OR = 4.16), nosocomial acquired bacteraemia (OR = 1.93), solid organ trasplantation (OR = 2.09) and shock (OR = 1.90). Independent risk factors associated with mortality in Klebsiella spp bacteraemia were: age (OR = 1.03), liver cirrhosis (OR = 2.63), ultimately or rapidly fatal prognosis of underlying disease (OR = 2.44), shock (OR = 8.60), pneumonia (OR = 4.96) or intraabdominal (OR = 3.85) source of bacteraemia and CTX-R isolate (OR = 4.63). Klebsiella spp is an important cause of bloodstream infection. CTX-R isolates have been increasing since 2000. CTX-R is an independent factor associated with mortality in Klebsiella spp bacteraemia.

  4. Isolation of Cronobacter spp. (Enterobacter Sakazakii) from Artisanal Mozzarella

    PubMed Central

    Rippa, Paola; Battaglia, Luciana; Parisi, Nicola

    2014-01-01

    Cronobacter spp. (Enterobacter sakazakii) is an opportunistic bacterial pathogen capable of causing disease and even fatalities in newborn infants within the first weeks of life if consumed as part of the diet. Premature and immunocompromised newborn infants are at particular risk. The microorganism has been isolated from a variety of foods including contaminated infant milk formula powder and milk powder substitute. The study aimed to evaluate the level of microbiological contamination in 47 samples of mozzarella cheese made with cow’s milk collected from artisan cheese producers in Southern Italy. Samples were collected from commercial sales points and underwent qualitative and quantitative microbiological analyses to test for the bacterial contaminants most commonly found in milk and cheese products. The 47 samples underwent qualitative and quantitative microbiological tests according to ISO UNI EN standards. Analyses focused on Staphylococcus aures, Salmonella spp., Listeria monocytogenes, Pseudomonas spp., E. coli, Yersinia spp., total coliforms and Cronobacter sakazakii. The ISO/TS 22964:2006 method was used to investigate possible contamination by C. sakazakii. Biochemical identification was carried out using an automated system for identification and susceptibility tests. None of the samples examined resulted positive for Salmonella spp. or Listeria spp. Only one sample resulted positive for Staphylococcus aureus. Pseudomonas spp. was isolated in 10 (21%) of 47 samples. High levels of total coliforms were found in 10 of 47 samples. Cronobacter spp. (Enterobacter sakazakii) was isolated in one sample. This is the first study to confirm isolation of C. sakazakii in artisan mozzarella cheese made from cow’s milk. The presence of C. sakazakii could be related to external contamination during the phases of production or to the use of contaminated milk. Since mozzarella is recommended in the diet of children and adults of all ages, this present study helps

  5. Radiosensitization of Salmonella spp. and Listeria spp. in ready-to-eat baby spinach leaves.

    PubMed

    Gomes, Carmen; Moreira, Rosana G; Castell-Perez, Elena

    2011-01-01

    The FDA recently approved irradiation treatment of leafy greens such as spinach up to 1 kGy; however, it is important to reduce the dose required to decontaminate the produce while maintaining its quality. Thus, the objectives of this study were: (1) to assess the radiation sensitivities of Salmonella spp. and Listeria spp. inoculated in ready-to-eat baby spinach leaves under modified atmosphere packaging (MAP) and irradiated using a 1.35-MeV Van de Graff accelerator (the leaves were irradiated both at room temperature and at -5 °C); and (2) to understand and optimize the synergistic effect of MAP and irradiation by studying the radiolysis of ozone formation under different temperatures, the effect of dose rate on its formation, and its decomposition. Results showed that increased concentrations of oxygen in the packaging significantly increased the radiation sensitivity of the test organisms, ranging from 7% up to 25% reduction in D(10)-values. In particular, radiosensitization could be effected (P < 0.05) by production of ozone, which increases with increasing dose-rate and oxygen concentration, and reducing temperatures. Radiosensitization was demonstrated for both microorganisms with irradiation of either fresh or frozen (-5 °C) baby spinach. These results suggest that low-dose (below 1 kGy) e-beam radiation under modified atmosphere packaging (100% O(2) and N(2):O(2)[1:1]) may be a viable tool for reducing microbial populations or eliminating Salmonella spp. and Listeria spp. from baby spinach. A suggested treatment to achieve a 5-log reduction of the test organisms would be irradiation at room temperature under 100% O(2) atmosphere at a dose level of 0.7 kGy. Practical Application: Decontamination of minimally processed fruits and vegetables from food-borne pathogens presents technical and economical challenges to the produce industry. Internalized microorganisms cannot be eliminated by the current procedure (water-washed or treated with 200-ppm chlorine

  6. Population dynamics of Aeromonas spp. in an urban river watershed.

    PubMed

    Pettibone, G W

    1998-10-01

    Density of Aeromonas spp. at one site in the Buffalo River and at four sites on its upstream tributaries was followed from June 1992-June 1993. Membrane filtration counts of Aeromonas during the summer ranged between 18 and 4000 ml-1, which were one to two logs higher than faecal coliform and faecal streptococci densities. Aeromonas spp. in the Buffalo River, and faecal coliforms, faecal streptococci, and the heterotrophic plate count throughout the watershed, increased by approximately one log during summer rainstorms. However, Aeromonas spp. increased only by a factor of two during rainstorms at the upstream sites. Aeromonas spp. showed a strong positive correlation with both indicator bacteria and total suspended solids at the upstream sites during the summer but not the winter. Correlations between Aeromonas and indicator bacteria remained strong in the Buffalo River during the winter, signifying that different conditions exist in the Buffalo River and its upstream tributaries. The strong correlation between Aeromonas spp. and indicator bacteria in the Buffalo River suggest that, in the absence of media capable of the quantitative recovery of potentially pathogenic aeromonads, standard faecal coliform analyses may adequately assess public health risks from Aeromonas spp. in an urban river used for recreational purposes.

  7. Chemical composition and antifungal potential of Brazilian propolis against Candida spp.

    PubMed

    Freires, I A; Queiroz, V C P P; Furletti, V F; Ikegaki, M; de Alencar, S M; Duarte, M C T; Rosalen, P L

    2016-06-01

    Propolis is known to have biological properties against numerous microorganisms of clinical interest. This study aimed to determine the chemical composition and antifungal activity of Brazilian propolis (types 3 and 13) against Candida spp. and their effects on the morphology of preformed and mature Candida biofilms. Samples of propolis (3 and 13) collected by Apis mellifera honeybees were obtained from different regions in Brazil. Ethanolic extracts of propolis (EEP) were prepared, fractionated and submitted to chemical analysis by GC/MS. The extracts and their hexane, dichloromethane and ethyl acetate fractions were tested for their ability to inhibit Candida spp. (C. albicans, C. dubliniensis, C. glabrata, C. kruzei, C. tropicalis and C. parapsilosis) by determination of the minimum inhibitory and fungicidal concentrations (MIC/MFC). Additionally, their effects on morphology of preformed and mature biofilms were observed by scanning electron microscopy. The phenolic compounds p-coumaric acid, caffeic acid phenethyl ester (CAPE), kaempferol and quercetin were identified in the EEP-3 and its bioactive dichloromethane fraction; and isoflavonoids such as medicarpin, vestitol and formononetin were found in the EEP-13, and triterpenes in its bioactive hexane fraction. The EEP-3 and EEP-13 and their bioactive fractions showed MIC values ranging from 0.2 to 125μg/mL and MFC values between 125 and 500μg/mL. The EEP and fractions were predominantly fungistatic agents. All extracts and fractions disrupted biofilm structures at 500μg/mL and amorphous areas with cell damage were clearly observed in preformed and mature biofilms. Propolis types 3 and 13 have strong anti-Candida activity and should be considered as promising candidates to treat oral and systemic candidiasis. PMID:26916845

  8. Exploration of natural enzyme inhibitors with hypoglycemic potentials amongst Eucalyptus Spp. by in vitro assays

    PubMed Central

    Dey, Baishakhi; Mitra, Analava; Katakam, Prakash; Singla, Rajeev K

    2014-01-01

    AIM: To investigate the presence and potency of natural enzyme inhibitors with hypoglycemic potentials amongst Eucalyptus Spp. by in vitro assays. METHODS: The leaf extracts of the three different Eucalyptus species [E. globulus (EG), E. citriodora (EC), E. camaldulensis (ECA)] were subjected to in vitro assay procedures to explore the prevalence of natural enzyme inhibitors (NEIs) after preliminary qualitative and quantitative phytochemical evaluations, to study their inhibitory actions against the enzymes like α-amylase, α-glucosidase, aldose reductase, angiotensin converting enzyme and dipeptidyl peptidase 4 playing pathogenic roles in type 2 diabetes. The antioxidant potential and total antioxidant capacity of the species were also evaluated. RESULTS: Major bioactive compounds like polyphenols (341.75 ± 3.63 to 496.85 ± 3.98) and flavonoids (4.89 ± 0.01 to 7.15 ± 0.02) were found in appreciable quantity in three species. Based on the IC50 values of the extracts under investigation, in all assays the effectivity was in the order of EG > ECA > EC. The results of the ferric reducing antioxidant power assay showed that the reducing ability of the species was also in the order of EG > ECA > EC. A strong correlation (R2 = 0.81-0.99) was found between the phenolic contents and the inhibitory potentials of the extracts against the targeted enzymes. CONCLUSION: These results show immense hypoglycemic potentiality of the Eucalyptus Spp. and a remarkable source of NEIs for a future phytotherapeutic approach in Type 2 diabetes. PMID:24748933

  9. Survey of thermotolerant Campylobacter spp. and Yersinia spp. in three surface water sources in Norway.

    PubMed

    Brennhovd, O; Kapperud, G; Langeland, G

    1992-01-01

    We investigated the occurrence of thermotolerant Campylobacter and Yersinia spp. in three surface water sources in Norway which represented different levels of pollution and eutrophication. Samples were collected every fortnight during a 14-month period. In addition, samples from 100 private wells were examined for campylobacters only. Campylobacter was recovered from 42 (43.8%) of the 96 samples of surface water, whereas Yersinia spp. were isolated from four (4.2%) of the samples. Campylobacter was not isolated from the well water samples. The highest isolation rate of Campylobacter was obtained from the two most polluted water sources. The proportion of positive samples was significantly higher in the autumn (71.4%) than in the spring (36.4%) or summer (22.2%). The highest overall isolation rate was obtained at water temperatures ranging from 2.1 to 8.0 degrees C, and the lowest at temperatures greater than 15 degrees C. Logistic regression analysis showed a highly significant relationship between the prevalence of Campylobacter and the number of three types of indicator bacteria: faecal coliforms, faecal streptococci and sulphite-reducing clostridia. Of the 60 Campylobacter isolates obtained, 51.7% belonged to C. jejuni biotype 1, 20.0% belonged to C. jejuni biotype 2, 21.7% to C. coli, 3.3% to C. lari and 3.3% were non-typable. All four Yersinia isolates were non-pathogenic variants.

  10. Burkholderia and Cupriavidus spp. are the preferred symbionts of Mimosa spp. in southern China.

    PubMed

    Liu, XiaoYun; Wei, Shuang; Wang, Fang; James, Euan K; Guo, XiaoYe; Zagar, Catherine; Xia, Liu Gui; Dong, Xin; Wang, Yi Peng

    2012-05-01

    Rhizobia were isolated from invasive Mimosa spp. (M. diplotricha and M. pudica) in Dehong district of the province of Yunnan in subtropical southern China. Almost all of the 98 isolates were β-rhizobia in the genera Burkholderia and Cupriavidus. These strains were analysed for their distribution characteristics together with strains from a previous study from Sishuangbanna. The proportion of nodules containing each β-rhizobial genus varied between Mimosa species, with Cupriavidus being predominant in M. diplotricha nodules (63.3% compared to 36.7% occupation with Burkholderia), but with M. pudica showing a slight preference for Burkholderia over Cupriavidus, with them occupying 56.5% and 43.5% of nodules, respectively. The symbiosis-essential genes nodA and nifH were present in all the Burkholderia and Cupriavidus strains tested, and their phylogenies indicated that these Mimosa symbionts share symbiotic genes with native South American rhizobia. The evolutionary discrepancies among 16S rRNA genes, nodA and nifH of Mimosa spp. symbionts, suggests that the nod and nif genes of β-rhizobia evolved independently.

  11. Desulfotomaculum spp.and Methanobacterium spp. Dominate a 4-5 km Deep Fault

    SciTech Connect

    Moser, Duane P.; Gihring, Thomas M.; Brockman, Fred J.; Fredrickson, Jim K.; Balkwill, David L.; Dollhopf, M E.; Lollar, B S.; Pratt, Lisa; Boice, E.; Southam, G; Wanger, Greg; Baker, Brett; Pfiffner, S; Lin, L; Onstott, T C.

    2005-12-01

    Sulfidic, 54-60 C, 3 to 30 million year old meteoric water stably Alkaline, sulfidic, 54 to 60 C, 4 to 53 million-year-old meteoric water emanating from a borehole intersecting quartzite-hosted fractures >3.3 km beneath the surface supported a microbial community dominated by a bacterial species affiliated with Desulfotomaculum spp. and an archaeal species related to Methanobacterium spp. The geochemical homogeneity over the 650-m length of the borehole, the lack of dividing cells, and the absence of these microorganisms in mine service water support an indigenous origin for the microbial community. The coexistence of these two microorganisms is consistent with a limiting flux of inorganic carbon and SO4 2 in the presence of high pH, high concentrations of H2 and CH4, and minimal free energy for autotrophic methanogenesis. Sulfide isotopic compositions were highly enriched, consistent with microbial SO4 2 reduction under hydrologic isolation. An analogous microbial couple and similar abiogenic gas chemistry have been reported recently for hydrothermal carbonate vents of the Lost City near the Mid-Atlantic Ridge (D. S. Kelly et al., Science 307:1428-1434, 2005), suggesting that these features may be common to deep subsurface habitats (continental and marine) bearing this geochemical signature. The geochemical setting and microbial communities described here are notably different from microbial ecosystems reported for shallower continental subsurface environments.

  12. [Infestation of Pseudopiazurus papayanus (Marshall) (Coleoptera: Curculionidae) on Carica spp. and Vasconcella spp. genotypes].

    PubMed

    Fancelli, Marilene; Sanches, Nilton F; Dantas, Jorge L L; Morales, Cinara F G; Caldas, Ranulfo C

    2008-01-01

    The papaya borer weevil, Pseudopiazurus papayanus (Marshall), is generally considered a secondary pest, but it has been reported in high infestations in Northeast Brazil. This work aimed at evaluating the occurrence of P. papayanus and reporting its infestation level in papaya genotypes kept at the germplasm bank of Embrapa Cassava & Tropical Fruits (Cruz das Almas, Bahia, Brazil). The number of larvae, pupae and adults found in each plant of 65 Carica spp. genotypes and of three Vasconcella spp. genotypes was registered in three to five plants of each genotype, by cutting the exsudating trunks lenghtwise. Papaya borer weevil was found in C. papaya and V. cauliflora but not in those of V. quercifolia. Among the evaluated genotypes, 52.4% of those belonging to the Solo group were infested, against 25.0% of the Formosa group. Larval infestation was the best criterion for sorting out genotypes concerning this insect infestation. This is also the first occurrence of the papaya borer weevil on V. cauliflora.

  13. Fluid extraction

    DOEpatents

    Wai, Chien M.; Laintz, Kenneth E.

    1999-01-01

    A method of extracting metalloid and metal species from a solid or liquid material by exposing the material to a supercritical fluid solvent containing a chelating agent is described. The chelating agent forms chelates that are soluble in the supercritical fluid to allow removal of the species from the material. In preferred embodiments, the extraction solvent is supercritical carbon dioxide and the chelating agent is a fluorinated .beta.-diketone. In especially preferred embodiments the extraction solvent is supercritical carbon dioxide, and the chelating agent comprises a fluorinated .beta.-diketone and a trialkyl phosphate, or a fluorinated .beta.-diketone and a trialkylphosphine oxide. Although a trialkyl phosphate can extract lanthanides and actinides from acidic solutions, a binary mixture comprising a fluorinated .beta.-diketone and a trialkyl phosphate or a trialkylphosphine oxide tends to enhance the extraction efficiencies for actinides and lanthanides. The method provides an environmentally benign process for removing contaminants from industrial waste without using acids or biologically harmful solvents. The method is particularly useful for extracting actinides and lanthanides from acidic solutions. The chelate and supercritical fluid can be regenerated, and the contaminant species recovered, to provide an economic, efficient process.

  14. Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil.

    PubMed

    Gottlieb, Juliana; André, Marcos Rogério; Soares, João Fábio; Gonçalves, Luiz Ricardo; Tonial de Oliveira, Mateus; Costa, Marcio Machado; Labruna, Marcelo Bahia; Bortolini, Carlos Eduardo; Machado, Rosangela Zacarias; Vieira, Maria Isabel Botelho

    2016-06-14

    Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) and Ehrlichia spp. (gene dsb). Five (8.6%) of the 58 dogs were serologically positive for Babesia spp. and three (5.1%) for E. canis. Four dogs (6.8%) were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed. PMID:27334817

  15. Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil.

    PubMed

    Gottlieb, Juliana; André, Marcos Rogério; Soares, João Fábio; Gonçalves, Luiz Ricardo; Tonial de Oliveira, Mateus; Costa, Marcio Machado; Labruna, Marcelo Bahia; Bortolini, Carlos Eduardo; Machado, Rosangela Zacarias; Vieira, Maria Isabel Botelho

    2016-06-14

    Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) and Ehrlichia spp. (gene dsb). Five (8.6%) of the 58 dogs were serologically positive for Babesia spp. and three (5.1%) for E. canis. Four dogs (6.8%) were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed. PMID:27304518

  16. Androgen Resistance in Squirrel Monkeys (Saimiri spp.)

    PubMed Central

    Gross, Katherine L; Westberry, Jenne M; Hubler, Tina R; Sadosky, Patti W; Singh, Ravinder J; Taylor, Robert L; Scammell, Jonathan G

    2008-01-01

    The goal of this study was to understand the basis for high androgen levels in squirrel monkeys (Saimiri spp.). Mass spectrometry was used to analyze serum testosterone, androstenedione, and dihydrotestosterone of male squirrel monkeys during the nonbreeding (n = 7) and breeding (n = 10) seasons. All hormone levels were elevated compared with those of humans, even during the nonbreeding season; the highest levels occurred during the breeding season. The ratio of testosterone to dihydrotestosterone in squirrel monkeys is high during the breeding season compared to man. Squirrel monkeys may have high testosterone to compensate for inefficient metabolism to dihydrotestosterone. We also investigated whether squirrel monkeys have high androgens to compensate for low-activity androgen receptors (AR). The response to dihydrotestosterone in squirrel monkey cells transfected with AR and AR-responsive reporter plasmids was 4-fold, compared with 28-fold in human cells. This result was not due to overexpression of cellular FKBP51, which causes glucocorticoid and progestin resistance in squirrel monkeys, because overexpression of FKBP51 had no effect on dihydrotestosterone-stimulated reporter activity in a human fibroblast cell line. To test whether the inherently low levels of FKBP52 in squirrel monkeys contribute to androgen insensitivity, squirrel monkey cells were transfected with an AR expression plasmid, an AR-responsive reporter plasmid, and a plasmid expressing FKBP52. Expression of FKBP52 decreased the EC50 or increased the maximal response to dihydrotestosterone. Therefore, the high androgen levels in squirrel monkeys likely compensate for their relatively low 5α-reductase activity during the breeding season and AR insensitivity resulting from low cellular levels of FKBP52. PMID:18724781

  17. Transcriptomic Signatures of Ash (Fraxinus spp.) Phloem

    PubMed Central

    Mamidala, Praveen; Bonello, Pierluigi; Herms, Daniel A.; Mittapalli, Omprakash

    2011-01-01

    Background Ash (Fraxinus spp.) is a dominant tree species throughout urban and forested landscapes of North America (NA). The rapid invasion of NA by emerald ash borer (Agrilus planipennis), a wood-boring beetle endemic to Eastern Asia, has resulted in the death of millions of ash trees and threatens billions more. Larvae feed primarily on phloem tissue, which girdles and kills the tree. While NA ash species including black (F. nigra), green (F. pennsylvannica) and white (F. americana) are highly susceptible, the Asian species Manchurian ash (F. mandshurica) is resistant to A. planipennis perhaps due to their co-evolutionary history. Little is known about the molecular genetics of ash. Hence, we undertook a functional genomics approach to identify the repertoire of genes expressed in ash phloem. Methodology and Principal Findings Using 454 pyrosequencing we obtained 58,673 high quality ash sequences from pooled phloem samples of green, white, black, blue and Manchurian ash. Intriguingly, 45% of the deduced proteins were not significantly similar to any sequences in the GenBank non-redundant database. KEGG analysis of the ash sequences revealed a high occurrence of defense related genes. Expression analysis of early regulators potentially involved in plant defense (i.e. transcription factors, calcium dependent protein kinases and a lipoxygenase 3) revealed higher mRNA levels in resistant ash compared to susceptible ash species. Lastly, we predicted a total of 1,272 single nucleotide polymorphisms and 980 microsatellite loci, among which seven microsatellite loci showed polymorphism between different ash species. Conclusions and Significance The current transcriptomic data provide an invaluable resource for understanding the genetic make-up of ash phloem, the target tissue of A. planipennis. These data along with future functional studies could lead to the identification/characterization of defense genes involved in resistance of ash to A. planipennis, and in future

  18. Butanol and ethanol production from tapioca starch wastewater by Clostridium spp.

    PubMed

    Ouephanit, Chanika; Virunanon, Chompunuch; Burapatana, Vorakan; Chulalaksananukul, Warawut

    2011-01-01

    Total (TWW) and tapioca starch wash wastewater (TSWW) from a cassava processing plant in Thailand were analyzed for their composition with a view to evaluate their potential as substrates for solvent production by ABE fermentation with Clostridium spp. Starch was detected at a 1.63-fold higher level in the TWW than that in the TSWW (24.4% and 15.0% (w/w), respectively). The chemical oxygen demand (COD) was broadly similar (20,093 and 20,433 mg/L), but the biological oxygen demand (BOD) was 1.84-fold higher (18,000 and 9,750 mg/L) in the TWW than that in the TSWW. Thus, the TSWW was selected as a substrate to evaluate its potential for butanol and ethanol production by two Clostridium spp. The combined ethanol plus butanol production in the TSWW at pH 6.5 was higher than that at pH 4.5, being around 27.8- and 3.4-fold higher in C. butyricum TISTR 1032 and C. acetobutylicum ATCC 824, respectively. In both strains, the butanol (and combined butanol plus ethanol) production level was improved at pH 5.5. The addition of yeast extract increased the bacterial cell production, but did not significantly improve solvent productivity in C. acetobutylicum, and even decreased butanol production by C. butyricum. PMID:22020468

  19. Biological Control of Phytophthora palmivora Causing Root Rot of Pomelo Using Chaetomium spp.

    PubMed Central

    Wattanachai, Pongnak; Kasem, Soytong; Poaim, Supatta

    2015-01-01

    Phytophthora diseases have become a major impediment in the citrus production in Thailand. In this study, an isolate of Phytophthora denominated as PHY02 was proven to be causal pathogen of root rot of Pomelo (Citrus maxima) in Thailand. The isolate PHY02 was morphologically characterized and identified as Phytophthora palmivora based on molecular analysis of an internal transcribed spacer rDNA sequence. This work also presents in vitro evaluations of the capacities of Chaetomium spp. to control the P. palmivora PHY02. As antagonists, Chaetomium globosum CG05, Chaetomium cupreum CC3003, Chaetomium lucknowense CL01 inhibited 50~61% mycelial growth, degraded mycelia and reduced 92~99% sporangial production of P. palmivora PHY02 in bi-culture test after 30 days. Fungal metabolites from Chaetomium spp. were tested against PHY02. Results showed that, methanol extract of C. globosum CG05 expressed strongest inhibitory effects on mycelial growth and sporangium formation of P. palmivora PHY02 with effective dose ED50 values of 26.5 µg/mL and 2.3 µg/mL, respectively. It is interesting that C. lucknowense is reported for the first time as an effective antagonist against a species of Phytophthora. PMID:25892917

  20. Evolutionary relationships among actinophages and a putative adaptation for growth in Streptomyces spp.

    PubMed

    Smith, Margaret C M; Hendrix, Roger W; Dedrick, Rebekah; Mitchell, Kaitlin; Ko, Ching-Chung; Russell, Daniel; Bell, Emma; Gregory, Matthew; Bibb, Maureen J; Pethick, Florence; Jacobs-Sera, Deborah; Herron, Paul; Buttner, Mark J; Hatfull, Graham F

    2013-11-01

    The genome sequences of eight Streptomyces phages are presented, four of which were isolated for this study. Phages R4, TG1, Hau3, and SV1 were isolated previously and have been exploited as tools for understanding and genetically manipulating Streptomyces spp. We also extracted five apparently intact prophages from recent Streptomyces spp. genome projects and, together with six phage genomes in the database, we analyzed all 19 Streptomyces phage genomes with a view to understanding their relationships to each other and to other actinophages, particularly the mycobacteriophages. Fifteen of the Streptomyces phages group into four clusters of related genomes. Although the R4-like phages do not share nucleotide sequence similarity with other phages, they clearly have common ancestry with cluster A mycobacteriophages, sharing many protein homologues, common gene syntenies, and similar repressor-stoperator regulatory systems. The R4-like phage Hau3 and the prophage StrepC.1 (from Streptomyces sp. strain C) appear to have hijacked a unique adaptation of the streptomycetes, i.e., use of the rare UUA codon, to control translation of the essential phage protein, the terminase. The Streptomyces venezuelae generalized transducing phage SV1 was used to predict the presence of other generalized transducing phages for different Streptomyces species.

  1. Butanol and ethanol production from tapioca starch wastewater by Clostridium spp.

    PubMed

    Ouephanit, Chanika; Virunanon, Chompunuch; Burapatana, Vorakan; Chulalaksananukul, Warawut

    2011-01-01

    Total (TWW) and tapioca starch wash wastewater (TSWW) from a cassava processing plant in Thailand were analyzed for their composition with a view to evaluate their potential as substrates for solvent production by ABE fermentation with Clostridium spp. Starch was detected at a 1.63-fold higher level in the TWW than that in the TSWW (24.4% and 15.0% (w/w), respectively). The chemical oxygen demand (COD) was broadly similar (20,093 and 20,433 mg/L), but the biological oxygen demand (BOD) was 1.84-fold higher (18,000 and 9,750 mg/L) in the TWW than that in the TSWW. Thus, the TSWW was selected as a substrate to evaluate its potential for butanol and ethanol production by two Clostridium spp. The combined ethanol plus butanol production in the TSWW at pH 6.5 was higher than that at pH 4.5, being around 27.8- and 3.4-fold higher in C. butyricum TISTR 1032 and C. acetobutylicum ATCC 824, respectively. In both strains, the butanol (and combined butanol plus ethanol) production level was improved at pH 5.5. The addition of yeast extract increased the bacterial cell production, but did not significantly improve solvent productivity in C. acetobutylicum, and even decreased butanol production by C. butyricum.

  2. Evolutionary Relationships among Actinophages and a Putative Adaptation for Growth in Streptomyces spp.

    PubMed Central

    Hendrix, Roger W.; Dedrick, Rebekah; Mitchell, Kaitlin; Ko, Ching-Chung; Russell, Daniel; Bell, Emma; Gregory, Matthew; Bibb, Maureen J.; Pethick, Florence; Jacobs-Sera, Deborah; Herron, Paul; Buttner, Mark J.; Hatfull, Graham F.

    2013-01-01

    The genome sequences of eight Streptomyces phages are presented, four of which were isolated for this study. Phages R4, TG1, ϕHau3, and SV1 were isolated previously and have been exploited as tools for understanding and genetically manipulating Streptomyces spp. We also extracted five apparently intact prophages from recent Streptomyces spp. genome projects and, together with six phage genomes in the database, we analyzed all 19 Streptomyces phage genomes with a view to understanding their relationships to each other and to other actinophages, particularly the mycobacteriophages. Fifteen of the Streptomyces phages group into four clusters of related genomes. Although the R4-like phages do not share nucleotide sequence similarity with other phages, they clearly have common ancestry with cluster A mycobacteriophages, sharing many protein homologues, common gene syntenies, and similar repressor-stoperator regulatory systems. The R4-like phage ϕHau3 and the prophage StrepC.1 (from Streptomyces sp. strain C) appear to have hijacked a unique adaptation of the streptomycetes, i.e., use of the rare UUA codon, to control translation of the essential phage protein, the terminase. The Streptomyces venezuelae generalized transducing phage SV1 was used to predict the presence of other generalized transducing phages for different Streptomyces species. PMID:23995638

  3. Multidrug-Resistant Acinetobacter spp.: Increasingly Problematic Nosocomial Pathogens

    PubMed Central

    Lee, Kyungwon; Yong, Dongeun; Jeong, Seok Hoon

    2011-01-01

    Pathogenic bacteria have increasingly been resisting to antimicrobial therapy. Recently, resistance problem has been relatively much worsened in Gram-negative bacilli. Acinetobacter spp. are typical nosocomial pathogens causing infections and high mortality, almost exclusively in compromised hospital patients. Acinetobacter spp. are intrinsically less susceptible to antibiotics than Enterobacteriaceae, and have propensity to acquire resistance. A surveillance study in Korea in 2009 showed that resistance rates of Acinetobacter spp. were very high: to fluoroquinolone 67%, to amikacin 48%, to ceftazidime 66% and to imipenem 51%. Carbapenem resistance was mostly due to OXA type carbapenemase production in A. baumannii isolates, whereas it was due to metallo-β-lactamase production in non-baumannii Acinetobacter isolates. Colistin-resistant isolates were rare but started to be isolated in Korea. Currently, the infection caused by multidrug-resistant A. baumannii is among the most difficult ones to treat. Analysis at tertiary care hospital in 2010 showed that among the 1,085 isolates of Acinetobacter spp., 14.9% and 41.8% were resistant to seven, and to all eight antimicrobial agents tested, respectively. It is known to be difficult to prevent Acinetobacter spp. infection in hospitalized patients, because the organisms are ubiquitous in hospital environment. Efforts to control resistant bacteria in Korea by hospitals, relevant scientific societies and government agencies have only partially been successful. We need concerted multidisciplinary efforts to preserve the efficacy of currently available antimicrobial agents, by following the principles of antimicrobial stewardship. PMID:22028150

  4. Thermophilic Campylobacter spp. in salad vegetables in Malaysia.

    PubMed

    Chai, Lay Ching; Robin, Tunung; Ragavan, Usha Menon; Gunsalam, Jurin Wolmon; Bakar, Fatimah Abu; Ghazali, Farinazleen Mohamad; Radu, Son; Kumar, Malakar Pradeep

    2007-06-10

    The main aim of this study was to combine the techniques of most probable number (MPN) and polymerase chain reaction (PCR) for quantifying the prevalence and numbers of Campylobacter spp. in ulam, a popular Malaysian salad dish, from a traditional wet market and two modern supermarkets in Selangor, Malaysia. A total of 309 samples of raw vegetables which are used in ulam were examined in the study. The prevalences of campylobacters in raw vegetables were, for supermarket I, Campylobacter spp., 51.9%; Campylobacter jejuni, 40.7%; and Campylobacter coli, 35.2%: for supermarket II, Campylobacter spp., 67.7%; C. jejuni, 67.7%; and C. coli, 65.7%: and for the wet market, Campylobacter spp., 29.4%; C. jejuni, 25.5%; and C. coli, 22.6%. In addition Campylobacter fetus was detected in 1.9% of raw vegetables from supermarket I. The maximum numbers of Campylobacter spp. in raw vegetables from supermarkets and the wet market were >2400 and 460 MPN/g, respectively.

  5. Protein Chips for Detection of Salmonella spp. from Enrichment Culture.

    PubMed

    Poltronieri, Palmiro; Cimaglia, Fabio; De Lorenzis, Enrico; Chiesa, Maurizio; Mezzolla, Valeria; Reca, Ida Barbara

    2016-01-01

    Food pathogens are the cause of foodborne epidemics, therefore there is a need to detect the pathogens in food productions rapidly. A pre-enrichment culture followed by selective agar plating are standard detection methods. Molecular methods such as qPCR have provided a first rapid protocol for detection of pathogens within 24 h of enrichment culture. Biosensors also may provide a rapid tool to individuate a source of Salmonella contamination at early times of pre-enrichment culture. Forty mL of Salmonella spp. enrichment culture were processed by immunoseparation using the Pathatrix, as in AFNOR validated qPCR protocols. The Salmonella biosensor combined with immunoseparation showed a limit of detection of 100 bacteria/40 mL, with a 400 fold increase to previous results. qPCR analysis requires processing of bead-bound bacteria with lysis buffer and DNA clean up, with a limit of detection of 2 cfu/50 μL. Finally, a protein chip was developed and tested in screening and identification of 5 common pathogen species, Salmonella spp., E. coli, S. aureus, Campylobacter spp. and Listeria spp. The protein chip, with high specificity in species identification, is proposed to be integrated into a Lab-on-Chip system, for rapid and reproducible screening of Salmonella spp. and other pathogen species contaminating food productions. PMID:27110786

  6. Protein Chips for Detection of Salmonella spp. from Enrichment Culture

    PubMed Central

    Poltronieri, Palmiro; Cimaglia, Fabio; De Lorenzis, Enrico; Chiesa, Maurizio; Mezzolla, Valeria; Reca, Ida Barbara

    2016-01-01

    Food pathogens are the cause of foodborne epidemics, therefore there is a need to detect the pathogens in food productions rapidly. A pre-enrichment culture followed by selective agar plating are standard detection methods. Molecular methods such as qPCR have provided a first rapid protocol for detection of pathogens within 24 h of enrichment culture. Biosensors also may provide a rapid tool to individuate a source of Salmonella contamination at early times of pre-enrichment culture. Forty mL of Salmonella spp. enrichment culture were processed by immunoseparation using the Pathatrix, as in AFNOR validated qPCR protocols. The Salmonella biosensor combined with immunoseparation showed a limit of detection of 100 bacteria/40 mL, with a 400 fold increase to previous results. qPCR analysis requires processing of bead-bound bacteria with lysis buffer and DNA clean up, with a limit of detection of 2 cfu/50 μL. Finally, a protein chip was developed and tested in screening and identification of 5 common pathogen species, Salmonella spp., E. coli, S. aureus, Campylobacter spp. and Listeria spp. The protein chip, with high specificity in species identification, is proposed to be integrated into a Lab-on-Chip system, for rapid and reproducible screening of Salmonella spp. and other pathogen species contaminating food productions. PMID:27110786

  7. [Human cryptosporidiosis and Cryptosporidium spp. in Haiti].

    PubMed

    Raccurt, Christian P; Brasseur, Philippe; Verdier, Rose I; Li, Xunde; Eyma, Etna; Stockman, Christine Pannier; Agnamey, Patrice; Guyot, Karine; Totet, Anne; Liautaud, Bernard; Nevez, Gilles; Dei-Cas, Eduardo; Pape, Jean W

    2006-06-01

    Contamination by water-born infectious diseases is closely linked to urban slums conditions such as overcrowding and high level of faecal pollution by animal and human excreta. In this environment, cryptosporidiosis is a major cause of acute diarrhoea in children and chronic persistent diarrhoea in AIDS patients, resulting in increased morbidity and mortality in both populations. The aims of this study conducted in Port-au-Prince, Haiti were to: (i) determine the frequency of Cryptosporidium infection in two populations of patients with diarrhoea, children and AIDS patients, and the existence of Cryptosporidium carriage in healthy adults living in close contact with them; (ii) identify by molecular genotyping the Cryptosporidium species involved; and (iii) evaluate the viability of Cryptosporidium oocysts isolated from human stools. From January 2000 to January 2001, 158 of 1529 diarrhoea stool samples collected from 93 patients with diarrhoea, 57 adults followed at Centres GHESKIO and 36 children admitted at the University Hospital in Port-au-Prince contained Cryptosporidium oocysts (10.3%). The majority of adult patients (98%) were HIV-infected whereas the majority of children (81%) tested negative for HIV. Cryptosporidium was documented in only 1/102 healthy persons living in contact with Cryptosporidium infected patients and infection was with the same genotype as that of the contact patient. Among the 69 Cryptosporidium isolates studied for genotyping, three species were identified: C. hominis (59%), C. parvum (38%) and C. felis (3%). The two C. felis cases are the first reported from AIDS patients in the Caribbean. Most of the children regardless of their HIV status were infected with C. hominis (72%), whereas AIDS patients were more likely to be infected by either human or animal genotypes. These data confirm that immunocompromised individuals are susceptible to a wide range of Cryptosporidium spp. Viability of Cryptosporidium oocysts were determined in an

  8. Extractant composition

    DOEpatents

    Smith, Barbara F.; Jarvinen, Gordon D.; Ryan, Robert R.

    1990-01-01

    An organic extracting solution useful for separating elements of the actinide series of the periodic table from elements of the lanthanide series, where both are in trivalent form. The extracting solution consists of a primary ligand and a secondary ligand, preferably in an organic solvent. The primary ligand is a substituted monothio-1,3-dicarbonyl, which includes a substituted 4-acyl-2-pyrazolin-5-thione, such as 4-benzoyl-2,4-dihydro-5-methyl-2-phenyl-3H-pyrazol-3-thione (BMPPT). The secondary ligand is a substituted phosphine oxide, such as trioctylphosphine oxide (TOPO).

  9. Simultaneously concentrating and pretreating of microalgae Chlorella spp. by three-phase partitioning.

    PubMed

    Li, Zhubo; Jiang, Feifei; Li, Ya; Zhang, Xu; Tan, Tianwei

    2013-12-01

    In this study, a recent simple separation technique, three-phase partitioning (TPP), was used for concentrating microalgae Chlorella spp. for the first time. More than 91.7% of the biomass precipitated in the interlayer of the system in 10 min. Temperature, initial concentration and ratio of ethanol to dipotassium hydrogen phosphate (DKP) were observed to negatively correlate with concentration factor while pH showed no significant influences. Using this method, biomass could be concentrated with much lower energy consumption and concentrated biomass could be conveniently collected. Besides, together with concentrating, TPP concentrated microalgae cells showed 26.3% increase in lipid extraction yield. Additionally, similarities in fatty acid profile indicated the avoidance of influence on lipid quality from chemicals. This study demonstrated the feasibility of TPP for microalgae biodiesel production. PMID:24121370

  10. Simultaneously concentrating and pretreating of microalgae Chlorella spp. by three-phase partitioning.

    PubMed

    Li, Zhubo; Jiang, Feifei; Li, Ya; Zhang, Xu; Tan, Tianwei

    2013-12-01

    In this study, a recent simple separation technique, three-phase partitioning (TPP), was used for concentrating microalgae Chlorella spp. for the first time. More than 91.7% of the biomass precipitated in the interlayer of the system in 10 min. Temperature, initial concentration and ratio of ethanol to dipotassium hydrogen phosphate (DKP) were observed to negatively correlate with concentration factor while pH showed no significant influences. Using this method, biomass could be concentrated with much lower energy consumption and concentrated biomass could be conveniently collected. Besides, together with concentrating, TPP concentrated microalgae cells showed 26.3% increase in lipid extraction yield. Additionally, similarities in fatty acid profile indicated the avoidance of influence on lipid quality from chemicals. This study demonstrated the feasibility of TPP for microalgae biodiesel production.

  11. Rapid and Sensitive Detection of Shigella spp. and Salmonella spp. by Multiple Endonuclease Restriction Real-Time Loop-Mediated Isothermal Amplification Technique

    PubMed Central

    Wang, Yi; Wang, Yan; Luo, Lijuan; Liu, Dongxin; Luo, Xia; Xu, Yanmei; Hu, Shoukui; Niu, Lina; Xu, Jianguo; Ye, Changyun

    2015-01-01

    Shigella and Salmonella are frequently isolated from various food samples and can cause human gastroenteritis. Here, a novel multiple endonuclease restriction real-time loop-mediated isothermal amplification technology (MERT-LAMP) were successfully established and validated for simultaneous detection of Shigella strains and Salmonella strains in only a single reaction. Two sets of MERT-LAMP primers for 2 kinds of pathogens were designed from ipaH gene of Shigella spp. and invA gene of Salmonella spp., respectively. Under the constant condition at 63°C, the positive results were yielded in as short as 12 min with the genomic DNA extracted from the 19 Shigella strains and 14 Salmonella strains, and the target pathogens present in a sample could be simultaneously identified based on distinct fluorescence curves in real-time format. Accordingly, the multiplex detection assay significantly reduced effort, materials and reagents used, and amplification and differentiation were conducted at the same time, obviating the use of postdetection procedures. The analytical sensitivity of MERT-LAMP was found to be 62.5 and 125 fg DNA/reaction with genomic templates of Shigella strains and Salmonella strains, which was consist with normal LAMP assay, and at least 10- and 100-fold more sensitive than that of qPCR and conventional PCR approaches. The limit of detection of MERT-LAMP for Shigella strains and Salmonella strains detection in artificially contaminated milk samples was 5.8 and 6.4 CFU per vessel. In conclusion, the MERT-LAMP methodology described here demonstrated a potential and valuable means for simultaneous screening of Shigella and Salmonella in a wide variety of samples. PMID:26697000

  12. Extractable resources

    NASA Technical Reports Server (NTRS)

    1975-01-01

    The use of information from space systems in the operation of extractive industries, particularly in exploration for mineral and fuel resources was reviewed. Conclusions and recommendations reported are based on the fundamental premise that survival of modern industrial society requires a continuing secure flow of resources for energy, construction and manufacturing, and for use as plant foods.

  13. Helminth fauna of Talpa spp. in the Palaearctic Realm.

    PubMed

    Ribas, A; Casanova, J C

    2006-03-01

    The helminth fauna of the genus Talpa in the Palaearctic Realm is reviewed. Several helminth species reported in Talpa spp. by a number of authors are discussed, with reference to host specificity, parasite biology, and host ethology, ecology and phylogeny. Twelve species of cestodes were found, two of which exhibit stenoxenous specificity (Staphylocystis bacillaris and Multitesticulata filamentosa). Only three species of trematodes, Ityogonimus lorum, Ityogonimus ocreatus and Combesia macrobursata, are exclusive parasites of Talpa spp. The largest group are nematodes, with 37 species. Species of Tricholinstowia are parasites of holarctic talpids and several species of distinct genera, such as Capillaria, Soboliphyme and Trichuris, are found only in Talpa spp. Only acanthocephalans of the genus Moniliformis have been reported in moles of the genus Talpa. On the basis of these helminthological findings, the close phylogenetic relationship between moles (Talpidae) and shrews (Soricidae) supports the separation of the ordinal levels Soricomorpha and Erinaceomorpha.

  14. Salmonellosis in laboratory-housed iguanid lizards (Sceloporus spp.).

    PubMed

    Kalvig, B A; Maggio-Price, L; Tsuji, J; Giddens, W E

    1991-10-01

    Fifteen wild-caught iguanid lizards (14 Sceloporus variabilis and one S. malachiticus) were used in a 3 mo study on thermal acclimation. Over a 2 mo period, five of the lizards showed decreased activity, anorexia and enlarged joints, and were either found moribund or were euthanatized due to their poor condition. Specimens taken from lesions in four of the five lizards were cultured and were infected with Salmonella spp. Salmonella spp. was cultured from cloacal swabs in six of the 10 surviving lizards. Standard metabolic rates of those that were infected did not differ significantly from those that were not infected. We postulate that the lizards were inapparent carriers of Salmonella spp. at the time of capture and, as a result of stress, five developed active overwhelming systemic infections. PMID:1758020

  15. SPPTOOLS: Programming tools for the IRAF SPP language

    NASA Technical Reports Server (NTRS)

    Fitzpatrick, M.

    1992-01-01

    An IRAF package to assist in SPP code development and debugging is described. SPP is the machine-independent programming language used by virtually all IRAF tasks. Tools have been written to aide both novice and advanced SPP programmers with development and debugging by providing tasks to check the code for the number and type of arguments in all calls to IRAF VOS library procedures, list the calling sequences of IRAF tasks, create a database of identifiers for quick access, check for memory which is not freed, and a source code formatter. Debugging is simplified since the programmer is able to get a better understanding of the structure of his/her code, and IRAF library procedure calls (probably the most common source of errors) are automatically checked for correctness.

  16. Identification of Novel Zoonotic Activity of Bartonella spp., France.

    PubMed

    Vayssier-Taussat, Muriel; Moutailler, Sara; Féménia, Françoise; Raymond, Philippe; Croce, Olivier; La Scola, Bernard; Fournier, Pierre-Edouard; Raoult, Didier

    2016-03-01

    Certain Bartonella species are known to cause afebrile bacteremia in humans and other mammals, including B. quintana, the agent of trench fever, and B. henselae, the agent of cat scratch disease. Reports have indicated that animal-associated Bartonella species may cause paucisymptomatic bacteremia and endocarditis in humans. We identified potentially zoonotic strains from 6 Bartonella species in samples from patients who had chronic, subjective symptoms and who reported tick bites. Three strains were B. henselae and 3 were from other animal-associated Bartonella spp. (B. doshiae, B. schoenbuchensis, and B. tribocorum). Genomic analysis of the isolated strains revealed differences from previously sequenced Bartonella strains. Our investigation identifed 3 novel Bartonella spp. strains with human pathogenic potential and showed that Bartonella spp. may be the cause of undifferentiated chronic illness in humans who have been bitten by ticks.

  17. Cadmium chloride susceptibility, a characteristic of Campylobacter spp.

    PubMed Central

    Kazmi, S U; Roberson, B S; Stern, N J

    1985-01-01

    We report a simple diagnostic characteristic useful in the presumptive identification of Campylobacter jejuni and Campylobacter coli. Filter paper disks impregnated with cadmium chloride were placed on streaked agar medium. Zones of growth inhibition for Campylobacter spp. occurred at 1.25 micrograms per disk. Other enteropathogens (Salmonella spp., Shigella spp., Vibrio cholerae, Vibrio parahaemolyticus, Escherichia coli, and Yersinia enterocolitica) were resistant to at least 40 micrograms per disk, with the exception of a strain of Shigella flexneri, which showed first susceptibility at 10 micrograms per disk. Most of the 52 Campylobacter strains, which were isolated from human clinical and animal sources, showed zones of inhibition greater than 10 mm with 2.5 micrograms of cadmium chloride per disk. At 20 micrograms per disk, Campylobacter isolates from clinical sources were significantly (P less than 0.01) more susceptible to cadmium chloride inhibition than were those from meat samples. Images PMID:3998099

  18. Identification of Novel Zoonotic Activity of Bartonella spp., France

    PubMed Central

    Vayssier-Taussat, Muriel; Moutailler, Sara; Féménia, Françoise; Raymond, Philippe; Croce, Olivier; La Scola, Bernard; Fournier, Pierre-Edouard

    2016-01-01

    Certain Bartonella species are known to cause afebrile bacteremia in humans and other mammals, including B. quintana, the agent of trench fever, and B. henselae, the agent of cat scratch disease. Reports have indicated that animal-associated Bartonella species may cause paucisymptomatic bacteremia and endocarditis in humans. We identified potentially zoonotic strains from 6 Bartonella species in samples from patients who had chronic, subjective symptoms and who reported tick bites. Three strains were B. henselae and 3 were from other animal-associated Bartonella spp. (B. doshiae, B. schoenbuchensis, and B. tribocorum). Genomic analysis of the isolated strains revealed differences from previously sequenced Bartonella strains. Our investigation identifed 3 novel Bartonella spp. strains with human pathogenic potential and showed that Bartonella spp. may be the cause of undifferentiated chronic illness in humans who have been bitten by ticks. PMID:26885624

  19. Survey and genetic characterization of wastewater in Tunisia for Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, Cyclospora cayetanensis and Eimeria spp.

    PubMed

    Ben Ayed, Layla; Yang, Wenli; Widmer, Giovanni; Cama, Vitaliano; Ortega, Ynes; Xiao, Lihua

    2012-09-01

    The microbial diversity of wastewater used for irrigation and fertilization was assessed using specific polymerase chain reaction (PCR) assays to detect and genotype several pathogenic protists including Cryptosporidium spp., Giardia duodenalis, Cyclospora spp., Eimeria spp. and Enterocytozoon bieneusi. A total of 220 wastewater samples (110 raw, 110 treated) and 12 sludge samples were collected from 2005 to 2008 from 18 treatment plants located throughout Tunisia. Except for Cyclospora, which was detected only once, E. bieneusi (61%), G. duodenalis (28%), Cryptosporidium spp. (27%) and Eimeria spp. (45%) were frequently observed in wastewater and sludge. Sequencing of PCR products showed that C. hominis, C. andersoni, G. duodenalis sub-assemblage A-II and E. bieneusi genotypes D and IV were the most prevalent. An analysis of the distribution of 209 internal transcribed spacer sequences of E. bieneusi originating from wastewater at the 18 treatment plants showed a similar genetic diversity, regardless of the geographical location. The identification of these parasite species and genotypes and of host-specific Eimeria species indicates that the microbial quality of wastewater was impacted by humans, livestock and rodents. Given the public health risks that some of these parasites represent, guidelines on wastewater usage are needed to minimize human exposure to these pathogens.

  20. OCCURRENCE OF Blastocystis spp. IN UBERABA, MINAS GERAIS, BRAZIL.

    PubMed

    Cabrine-Santos, Marlene; Cintra, Eduardo do Nascimento; do Carmo, Rafaela Andrade; Nascentes, Gabriel Antônio Nogueira; Pedrosa, André Luiz; Correia, Dalmo; Oliveira-Silva, Márcia Benedita de

    2015-01-01

    Intestinal parasites are a problem for public health all over the world. The infection with Blastocystis, a protozoan of controversial pathogenicity, is one of the most common among them all. In this study, the occurrence of intestinal parasites, with emphasis on Blastocystis, in patients at the Universidade Federal do Triângulo Mineiro was investigated in Uberaba (MG) through microscopy of direct smears and fecal concentrates using Ritchie's method. Feces of 1,323 patients were examined from April 2011 to May 2012. In 28.7% of them at least one intestinal parasite was identified, and the most frequent organisms were Blastocystis spp. (17.8%) and Giardia intestinalis (7.4%). The occurrence of parasitism was higher in children aged 6 -10 years old, and the infection with Blastocystis spp. was higher above the age of six (p < 0.001). The exclusive presence of G. intestinalis and of Blastocystis spp. was observed in 5.4% and 12.2% of the patients, respectively. Regarding patients with diarrheic feces, 8% revealed unique parasitism of Blastocystis spp. Other intestinal parasites observed in children were Ascaris lumbricoides (0.3%) and Entamoeba histolytica/dispar/moshkovskii (1.4%). The Ritchie's method was more sensitive (92.8%) when compared to direct microscopy (89.8%), with high agreement between them (97.7%, kappa = 0.92). In conclusion, the occurrence of Blastocystis spp. in Uberaba is high and the presence of diarrheic feces with exclusive presence of the parasite of Blastocystis spp. was observed. PMID:26200960

  1. OCCURRENCE OF Blastocystis spp. IN UBERABA, MINAS GERAIS, BRAZIL

    PubMed Central

    CABRINE-SANTOS, Marlene; CINTRA, Eduardo do Nascimento; do CARMO, Rafaela Andrade; NASCENTES, Gabriel Antônio Nogueira; PEDROSA, André Luiz; CORREIA, Dalmo; de OLIVEIRA-SILVA, Márcia Benedita

    2015-01-01

    Intestinal parasites are a problem for public health all over the world. The infection with Blastocystis, a protozoan of controversial pathogenicity, is one of the most common among them all. In this study, the occurrence of intestinal parasites, with emphasis on Blastocystis, in patients at the Universidade Federal do Triângulo Mineiro was investigated in Uberaba (MG) through microscopy of direct smears and fecal concentrates using Ritchie’s method. Feces of 1,323 patients were examined from April 2011 to May 2012. In 28.7% of them at least one intestinal parasite was identified, and the most frequent organisms were Blastocystis spp. (17.8%) and Giardia intestinalis (7.4%). The occurrence of parasitism was higher in children aged 6 -10 years old, and the infection with Blastocystis spp. was higher above the age of six (p < 0.001). The exclusive presence of G. intestinalis and of Blastocystis spp. was observed in 5.4% and 12.2% of the patients, respectively. Regarding patients with diarrheic feces, 8% revealed unique parasitism of Blastocystis spp. Other intestinal parasites observed in children were Ascaris lumbricoides(0.3%) and Entamoeba histolytica/dispar/moshkovskii (1.4%). The Ritchie’s method was more sensitive (92.8%) when compared to direct microscopy (89.8%), with high agreement between them (97.7%, kappa = 0.92). In conclusion, the occurrence of Blastocystis spp. in Uberaba is high and the presence of diarrheic feces with exclusive presence of the parasite of Blastocystis spp. was observed. PMID:26200960

  2. Laboratory diagnosis of Blastocystis spp. in diarrheic patients

    PubMed Central

    Elghareeb, Azza S; Younis, Mohamed S; El Fakahany, Amany F; Nagaty, Ibrahim M; Nagib, Marwa M

    2015-01-01

    Background: Many laboratories currently diagnose Blastocystis spp. infections by looking for the presence of vacuolar forms in faeces and the amoeboid form in diarrheal stools. Objectives: To investigate the best direct method in diagnosis of Blastocystis spp. and to study different morphological forms of the parasite. Materials and Methods: The study was carried out on one thousand and two hundred diarrheic stool samples. All samples were examined using direct smear, iodine stained smear, formalin-ether concentration techniques, trichrome stained smear and in vitro cultivation using Jones' medium. Results: Using direct smear, Blastocystis spp was detected in 42 cases (3.5%) with a sensitivity (28.4%) and specificity (100%). Iodine stained smear detected 72 positive cases (6%) with a sensitivity (48.7%), specificity (100%). Formol ether concentration technique detected 120 positive cases (10%) with a sensitivity (81.1%) and specificity (100%). Trichrome stained smear detected 148 positive cases (12.3%). In vitro cultivation using Joni's medium detected 274 positive cases (22.8%) which was the highest number among all different diagnostic methods with a sensitivity (100%) ,specificity (88%), PPV (54.1%) and NPV (100%). It was found that, 49 blastocystosis cases had mixed infection with other intestinal parasites. Giardia lamblia was the most frequently associated parasite with Blastocystis spp. Conclusion: In vitro cultivation is more sensitive in detection of B. hominis than simple smear and concentration technique. Blastocystis spp. vacuolar form was the most common form that was found by all methods used in this study G. lamblia was the most frequent parasite associated with Blastocystis spp . PMID:25709951

  3. An ectopic case of Tunga spp. infection in Peru.

    PubMed

    Maco, Vicente; Maco, Vicente P; Gotuzzo, Eduardo

    2010-06-01

    Tungiasis is a neglected ectoparasitism of impoverished areas in South America and sub-Saharan Africa. The sand flea Tunga spp. preferably infests the soles and the periungueal and interdigital regions of the feet. Ectopic tungiasis is rare, even in highly endemic areas. We describe a case of an indigenous patient in Peru who presented with a nodular lesion in the extensor aspect of the knee and whose biopsy was compatible with Tunga spp. This is the first documented case of knee tungiasis in an endemic country. The historical, clinical, histological, and current epidemiological aspects of tungiasis in Peru are discussed here. PMID:20519602

  4. An Ectopic Case of Tunga spp. Infection in Peru

    PubMed Central

    Maco, Vicente; Maco, Vicente P.; Gotuzzo, Eduardo

    2010-01-01

    Tungiasis is a neglected ectoparasitism of impoverished areas in South America and sub-Saharan Africa. The sand flea Tunga spp. preferably infests the soles and the periungueal and interdigital regions of the feet. Ectopic tungiasis is rare, even in highly endemic areas. We describe a case of an indigenous patient in Peru who presented with a nodular lesion in the extensor aspect of the knee and whose biopsy was compatible with Tunga spp. This is the first documented case of knee tungiasis in an endemic country. The historical, clinical, histological, and current epidemiological aspects of tungiasis in Peru are discussed here. PMID:20519602

  5. Variables Associated with Infections of Cattle by Brucella abortus., Leptospira spp. and Neospora spp. in Amazon Region in Brazil.

    PubMed

    Chiebao, D P; Valadas, S Y O B; Minervino, A H H; Castro, V; Romaldini, A H C N; Calhau, A S; De Souza, R A B; Gennari, S M; Keid, L B; Soares, R M

    2015-10-01

    The frequency of Neospora spp., Leptospira spp. and Brucella abortus infections in adult cattle was determined in herds of the State of Pará, Brazil, which is an important region for cattle production located in the Amazon region. A total of 3466 adult female cattle from 176 herds were tested, leading to a frequency of seropositive animals of 14.7%, 3.7% and 65.5% and a herd positivity of 87.4%, 41.3% and 98.8% for infections caused by Neospora spp., B. abortus and Leptospira spp., respectively. The five most frequently diagnosed serologic responses to Leptospira spp. were those against serovars hardjo, wolfii, grippotyphosa, hebdomadis and shermani. The following associations were found: practice of artificial insemination, large farm size, large herd size, large number of dogs and high number of total abortions per year with the presence of antibodies against serovar hardjo; positive results to serovar grippotyphosa with the presence of dogs; inappropriate disposal of aborted foetuses with positivity to serovar hebdomadis. Serovar grippotyphosa was also associated with number of episodes of abortions. Neospora spp. positive herds were associated with episodes of abortion and B. abortus infection with the disposal of dead animals and aborted foetuses on pastures and with the use of artificial insemination. In conclusion, the high frequency of brucellosis, leptospirosis and neosporosis in the region may be a consequence of social, natural and raising conditions as: (i) climate conditions that favour the survival and spread of pathogens in the environment; (ii) farms located in regions bordering forest areas; (iii) farms in areas of difficult access to the veterinary service; (iv) extensive beef herds raised at pastures with different age and productive groups inter-mingled; and (v) minimal concerns regarding hygiene practices and disease prevention measures. PMID:26302373

  6. Canine infection with Dirofilaria immitis, Borrelia burgdorferi, Anaplasma spp., and Ehrlichia spp. in the United States, 2010–2012

    PubMed Central

    2014-01-01

    Background The geographic distribution of canine infection with vector-borne disease agents in the United States appears to be expanding. Methods To provide an updated assessment of geographic trends in canine infection with Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia spp., and Anaplasma spp., we evaluated results from an average of 3,588,477 dogs tested annually by veterinarians throughout the United States from 2010 – 2012. Results As in an earlier summary report, the percent positive test results varied by agent and region, with antigen of D. immitis and antibody to Ehrlichia spp. most commonly identified in the Southeast (2.9% and 3.2%, respectively) and antibody to both B. burgdorferi and Anaplasma spp. most commonly identified in the Northeast (13.3% and 7.1%, respectively) and upper Midwest (4.4% and 3.9%, respectively). Percent positive test results for D. immitis antigen were lower in every region considered, including in the Southeast, than previously reported. Percent positive test results for antibodies to B. burgdorferi and Ehrlichia spp. were higher nationally than previously reported, and, for antibodies to Anaplasma spp., were higher in the Northeast but lower in the Midwest and West, than in the initial report. Annual reports of human cases of Lyme disease, ehrlichiosis, and anaplasmosis were associated with percent positive canine test results by state for each respective tick-borne disease agent (R2 = 0.701, 0.457, and 0.314, respectively). Within endemic areas, percent positive test results for all three tick-borne agents demonstrated evidence of geographic expansion. Conclusions Continued national monitoring of canine test results for vector-borne zoonotic agents is an important tool for accurately mapping the geographic distribution of these agents, and greatly aids our understanding of the veterinary and public health threats they pose. PMID:24886589

  7. Variables Associated with Infections of Cattle by Brucella abortus., Leptospira spp. and Neospora spp. in Amazon Region in Brazil.

    PubMed

    Chiebao, D P; Valadas, S Y O B; Minervino, A H H; Castro, V; Romaldini, A H C N; Calhau, A S; De Souza, R A B; Gennari, S M; Keid, L B; Soares, R M

    2015-10-01

    The frequency of Neospora spp., Leptospira spp. and Brucella abortus infections in adult cattle was determined in herds of the State of Pará, Brazil, which is an important region for cattle production located in the Amazon region. A total of 3466 adult female cattle from 176 herds were tested, leading to a frequency of seropositive animals of 14.7%, 3.7% and 65.5% and a herd positivity of 87.4%, 41.3% and 98.8% for infections caused by Neospora spp., B. abortus and Leptospira spp., respectively. The five most frequently diagnosed serologic responses to Leptospira spp. were those against serovars hardjo, wolfii, grippotyphosa, hebdomadis and shermani. The following associations were found: practice of artificial insemination, large farm size, large herd size, large number of dogs and high number of total abortions per year with the presence of antibodies against serovar hardjo; positive results to serovar grippotyphosa with the presence of dogs; inappropriate disposal of aborted foetuses with positivity to serovar hebdomadis. Serovar grippotyphosa was also associated with number of episodes of abortions. Neospora spp. positive herds were associated with episodes of abortion and B. abortus infection with the disposal of dead animals and aborted foetuses on pastures and with the use of artificial insemination. In conclusion, the high frequency of brucellosis, leptospirosis and neosporosis in the region may be a consequence of social, natural and raising conditions as: (i) climate conditions that favour the survival and spread of pathogens in the environment; (ii) farms located in regions bordering forest areas; (iii) farms in areas of difficult access to the veterinary service; (iv) extensive beef herds raised at pastures with different age and productive groups inter-mingled; and (v) minimal concerns regarding hygiene practices and disease prevention measures.

  8. Invasion Assays and Genomotyping to Investigate Differences in Virulence of Campylobacter spp. Isolates from Iceland

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter spp. are the leading cause of human gastroenteritis worldwide. Epithelial cell invasion is thought to be essential for Campylobacter spp. infection. Previous invasion studies with intestinal epithelial cells revealed that the ability of different Campylobacter jejuni isolates to inva...

  9. URANIUM EXTRACTION

    DOEpatents

    Harrington, C.D.; Opie, J.V.

    1958-07-01

    The recovery of uranium values from uranium ore such as pitchblende is described. The ore is first dissolved in nitric acid, and a water soluble nitrate is added as a salting out agent. The resulting feed solution is then contacted with diethyl ether, whereby the bulk of the uranyl nitrate and a portion of the impurities are taken up by the ether. This acid ether extract is then separated from the aqueous raffinate, and contacted with water causing back extractioa of the uranyl nitrate and impurities into the water to form a crude liquor. After separation from the ether extract, this crude liquor is heated to about 118 deg C to obtain molten uranyl nitrate hexahydratc. After being slightly cooled the uranyl nitrate hexahydrate is contacted with acid free diethyl ether whereby the bulk of the uranyl nitrate is dissolved into the ethcr to form a neutral ether solution while most of the impurities remain in the aqueous waste. After separation from the aqueous waste, the resultant ether solution is washed with about l0% of its volume of water to free it of any dissolved impurities and is then contacted with at least one half its volume of water whereby the uranyl nitrate is extracted into the water to form an aqueous product solution.

  10. Characterization of Lavandula spp. Honey Using Multivariate Techniques

    PubMed Central

    2016-01-01

    Traditionally, melissopalynological and physicochemical analyses have been the most used to determine the botanical origin of honey. However, when performed individually, these analyses may provide less unambiguous results, making it difficult to discriminate between mono and multifloral honeys. In this context, with the aim of better characterizing this beehive product, a selection of 112 Lavandula spp. monofloral honey samples from several regions were evaluated by association of multivariate statistical techniques with physicochemical, melissopalynological and phenolic compounds analysis. All honey samples fulfilled the quality standards recommended by international legislation, except regarding sucrose content and diastase activity. The content of sucrose and the percentage of Lavandula spp. pollen have a strong positive association. In fact, it was found that higher amounts of sucrose in honey are related with highest percentage of pollen of Lavandula spp.. The samples were very similar for most of the physicochemical parameters, except for proline, flavonoids and phenols (bioactive factors). Concerning the pollen spectrum, the variation of Lavandula spp. pollen percentage in honey had little contribution to the formation of samples groups. The formation of two groups regarding the physicochemical parameters suggests that the presence of other pollen types in small percentages influences the factor termed as “bioactive”, which has been linked to diverse beneficial health effects. PMID:27588420

  11. A SURVEY OF CYST NEMATODES (HETERODERA SPP.) IN NORTHERN EGYPT

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Information concerning the occurrence and distribution of cyst nematodes (Heterodera spp.) in Egypt is important to assess their potential to cause economic damage to crop plants. A nematode survey was conducted in Alexandria and El-Behera Governorates in northern Egypt to identify the species of cy...

  12. 21 CFR 866.3630 - Serratia spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Serratia spp. serological reagents. 866.3630 Section 866.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3630 Serratia...

  13. 21 CFR 866.3660 - Shigella spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Shigella spp. serological reagents. 866.3660 Section 866.3660 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3660 Shigella...

  14. 21 CFR 866.3660 - Shigella spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Shigella spp. serological reagents. 866.3660 Section 866.3660 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3660 Shigella...

  15. 21 CFR 866.3630 - Serratia spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Serratia spp. serological reagents. 866.3630 Section 866.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3630 Serratia...

  16. The copepod Calanus spp. (Calanidae) is repelled by polarized light

    PubMed Central

    Lerner, Amit; Browman, Howard I.

    2016-01-01

    Both attraction and repulsion from linearly polarized light have been observed in zooplankton. A dichotomous choice experiment, consisting of plankton light traps deployed in natural waters at a depth of 30 m that projected either polarized or unpolarized light of the same intensity, was used to test the hypothesis that the North Atlantic copepod, Calanus spp., is linearly polarotactic. In addition, the transparency of these copepods, as they might be seen by polarization insensitive vs. sensitive visual systems, was measured. Calanus spp. exhibited negative polarotaxis with a preference ratio of 1.9:1. Their transparency decreased from 80% to 20% to 30% in the unpolarized, partially polarized, and electric (e-) vector orientation domains respectively - that is, these copepods would appear opaque and conspicuous to a polarization-sensitive viewer looking at them under conditions rich in polarized light. Since the only difference between the two plankton traps was the polarization cue, we conclude that Calanus spp. are polarization sensitive and exhibit negative polarotaxis at low light intensities (albeit well within the sensitivity range reported for copepods). We hypothesize that Calanus spp. can use polarization vision to reduce their risk of predation by polarization-sensitive predators and suggest that this be tested in future experiments. PMID:27762400

  17. Inactivation of Salmonella spp. on tomatoes by plant molecules

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The efficacy of carvacrol (CAR), trans-cinnamaldehyde (TC), eugenol (EUG) and ß-resorcylic acid (BR) as a wash treatment for reducing Salmonella spp. on tomatoes was investigated. Plum tomatoes inoculated with a six-serotype mixture of Salmonella (108 CFU) were subjected to washing in sterile deion...

  18. Characterization of Lavandula spp. Honey Using Multivariate Techniques.

    PubMed

    Estevinho, Leticia M; Chambó, Emerson Dechechi; Pereira, Ana Paula Rodrigues; Carvalho, Carlos Alfredo Lopes de; Toledo, Vagner de Alencar Arnaut de

    2016-01-01

    Traditionally, melissopalynological and physicochemical analyses have been the most used to determine the botanical origin of honey. However, when performed individually, these analyses may provide less unambiguous results, making it difficult to discriminate between mono and multifloral honeys. In this context, with the aim of better characterizing this beehive product, a selection of 112 Lavandula spp. monofloral honey samples from several regions were evaluated by association of multivariate statistical techniques with physicochemical, melissopalynological and phenolic compounds analysis. All honey samples fulfilled the quality standards recommended by international legislation, except regarding sucrose content and diastase activity. The content of sucrose and the percentage of Lavandula spp. pollen have a strong positive association. In fact, it was found that higher amounts of sucrose in honey are related with highest percentage of pollen of Lavandula spp.. The samples were very similar for most of the physicochemical parameters, except for proline, flavonoids and phenols (bioactive factors). Concerning the pollen spectrum, the variation of Lavandula spp. pollen percentage in honey had little contribution to the formation of samples groups. The formation of two groups regarding the physicochemical parameters suggests that the presence of other pollen types in small percentages influences the factor termed as "bioactive", which has been linked to diverse beneficial health effects. PMID:27588420

  19. 21 CFR 866.3630 - Serratia spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Serratia spp. serological reagents. 866.3630 Section 866.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3630 Serratia...

  20. 21 CFR 866.3630 - Serratia spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Serratia spp. serological reagents. 866.3630 Section 866.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3630 Serratia...

  1. 21 CFR 866.3660 - Shigella spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Shigella spp. serological reagents. 866.3660 Section 866.3660 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3660 Shigella...

  2. 21 CFR 866.3660 - Shigella spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Shigella spp. serological reagents. 866.3660 Section 866.3660 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3660 Shigella...

  3. 21 CFR 866.3660 - Shigella spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Shigella spp. serological reagents. 866.3660 Section 866.3660 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3660 Shigella...

  4. 21 CFR 866.3630 - Serratia spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Serratia spp. serological reagents. 866.3630 Section 866.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3630 Serratia...

  5. Bordetella pseudohinzii spp. nov. infects C57Bl6 mice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clinical studies rely heavily on mouse models of infection. Precise identification and control of contaminating pathogens that circulate in mouse colonies is an important task. Over the past decade, there have been several reports documenting the isolation of Bordetella spp. from purported pathog...

  6. Distribution of Anastrepha spp. in carambola orchards: Evidence for migration

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Carambola orchards in Juana Diaz, Corozal, and Isabela, PR, were monitored for Anastrepha spp. fruit flies using Multi-lure traps baited with putrescine and ammonium acetate. The number of flies at various locations within the orchards were statistically compared with the expected distribution if fl...

  7. 21 CFR 866.3300 - Haemophilus spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Haemophilus spp. serological reagents. 866.3300 Section 866.3300 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3300...

  8. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200...

  9. 21 CFR 866.3340 - Klebsiella spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Klebsiella spp. serological reagents. 866.3340 Section 866.3340 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3340...

  10. 21 CFR 866.3125 - Citrobacter spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Citrobacter spp. serological reagents. 866.3125 Section 866.3125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3125...

  11. 21 CFR 866.3125 - Citrobacter spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Citrobacter spp. serological reagents. 866.3125 Section 866.3125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3125...

  12. 21 CFR 866.3085 - Brucella spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Brucella spp. serological reagents. 866.3085 Section 866.3085 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3085 Brucella...

  13. 21 CFR 866.3415 - Pseudomonas spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Pseudomonas spp. serological reagents. 866.3415 Section 866.3415 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3415...

  14. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  15. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

  16. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200...

  17. 21 CFR 866.3340 - Klebsiella spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Klebsiella spp. serological reagents. 866.3340 Section 866.3340 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3340...

  18. 21 CFR 866.3040 - Aspergillus spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Aspergillus spp. serological reagents. 866.3040 Section 866.3040 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3040...

  19. 21 CFR 866.3300 - Haemophilus spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Haemophilus spp. serological reagents. 866.3300 Section 866.3300 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3300...

  20. 21 CFR 866.3125 - Citrobacter spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Citrobacter spp. serological reagents. 866.3125 Section 866.3125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3125...

  1. 21 CFR 866.3085 - Brucella spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Brucella spp. serological reagents. 866.3085 Section 866.3085 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3085 Brucella...

  2. 21 CFR 866.3085 - Brucella spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Brucella spp. serological reagents. 866.3085 Section 866.3085 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3085 Brucella...

  3. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  4. 21 CFR 866.3350 - Leptospira spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Leptospira spp. serological reagents. 866.3350 Section 866.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3350...

  5. 21 CFR 866.3415 - Pseudomonas spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Pseudomonas spp. serological reagents. 866.3415 Section 866.3415 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3415...

  6. 21 CFR 866.3350 - Leptospira spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Leptospira spp. serological reagents. 866.3350 Section 866.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3350...

  7. 21 CFR 866.3085 - Brucella spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Brucella spp. serological reagents. 866.3085 Section 866.3085 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3085 Brucella...

  8. 21 CFR 866.3270 - Flavobacterium spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Flavobacterium spp. serological reagents. 866.3270 Section 866.3270 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  9. 21 CFR 866.3340 - Klebsiella spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Klebsiella spp. serological reagents. 866.3340 Section 866.3340 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3340...

  10. 21 CFR 866.3415 - Pseudomonas spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Pseudomonas spp. serological reagents. 866.3415 Section 866.3415 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3415...

  11. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  12. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

  13. 21 CFR 866.3040 - Aspergillus spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Aspergillus spp. serological reagents. 866.3040 Section 866.3040 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3040...

  14. 21 CFR 866.3355 - Listeria spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Listeria spp. serological reagents. 866.3355 Section 866.3355 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3355 Listeria...

  15. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200...

  16. 21 CFR 866.3270 - Flavobacterium spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Flavobacterium spp. serological reagents. 866.3270 Section 866.3270 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  17. 21 CFR 866.3350 - Leptospira spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Leptospira spp. serological reagents. 866.3350 Section 866.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3350...

  18. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  19. 21 CFR 866.3340 - Klebsiella spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Klebsiella spp. serological reagents. 866.3340 Section 866.3340 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3340...

  20. 21 CFR 866.3140 - Corynebacterium spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Corynebacterium spp. serological reagents. 866.3140 Section 866.3140 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  1. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

  2. 21 CFR 866.3125 - Citrobacter spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Citrobacter spp. serological reagents. 866.3125 Section 866.3125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3125...

  3. 21 CFR 866.3270 - Flavobacterium spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Flavobacterium spp. serological reagents. 866.3270 Section 866.3270 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  4. 21 CFR 866.3355 - Listeria spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Listeria spp. serological reagents. 866.3355 Section 866.3355 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3355 Listeria...

  5. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

  6. 21 CFR 866.3270 - Flavobacterium spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Flavobacterium spp. serological reagents. 866.3270 Section 866.3270 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  7. 21 CFR 866.3355 - Listeria spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Listeria spp. serological reagents. 866.3355 Section 866.3355 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3355 Listeria...

  8. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200...

  9. 21 CFR 866.3140 - Corynebacterium spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Corynebacterium spp. serological reagents. 866.3140 Section 866.3140 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  10. 21 CFR 866.3035 - Arizona spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Arizona spp. serological reagents. 866.3035 Section 866.3035 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3035 Arizona...

  11. 21 CFR 866.3040 - Aspergillus spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Aspergillus spp. serological reagents. 866.3040 Section 866.3040 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3040...

  12. 21 CFR 866.3140 - Corynebacterium spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Corynebacterium spp. serological reagents. 866.3140 Section 866.3140 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  13. 21 CFR 866.3355 - Listeria spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Listeria spp. serological reagents. 866.3355 Section 866.3355 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3355 Listeria...

  14. 21 CFR 866.3040 - Aspergillus spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Aspergillus spp. serological reagents. 866.3040 Section 866.3040 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3040...

  15. 21 CFR 866.3350 - Leptospira spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Leptospira spp. serological reagents. 866.3350 Section 866.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3350...

  16. 21 CFR 866.3140 - Corynebacterium spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Corynebacterium spp. serological reagents. 866.3140 Section 866.3140 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  17. 21 CFR 866.3415 - Pseudomonas spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Pseudomonas spp. serological reagents. 866.3415 Section 866.3415 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3415...

  18. 21 CFR 866.3035 - Arizona spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Arizona spp. serological reagents. 866.3035 Section 866.3035 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3035 Arizona...

  19. 21 CFR 866.3035 - Arizona spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Arizona spp. serological reagents. 866.3035 Section 866.3035 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3035 Arizona...

  20. 21 CFR 866.3300 - Haemophilus spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Haemophilus spp. serological reagents. 866.3300 Section 866.3300 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3300...

  1. 21 CFR 866.3300 - Haemophilus spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Haemophilus spp. serological reagents. 866.3300 Section 866.3300 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3300...

  2. 21 CFR 866.3035 - Arizona spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Arizona spp. serological reagents. 866.3035 Section 866.3035 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3035 Arizona...

  3. 21 CFR 866.3035 - Arizona spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Arizona spp. serological reagents. 866.3035 Section 866.3035 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3035 Arizona...

  4. 21 CFR 866.3085 - Brucella spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Brucella spp. serological reagents. 866.3085 Section 866.3085 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3085 Brucella...

  5. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

  6. 21 CFR 866.3140 - Corynebacterium spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Corynebacterium spp. serological reagents. 866.3140 Section 866.3140 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  7. 21 CFR 866.3355 - Listeria spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Listeria spp. serological reagents. 866.3355 Section 866.3355 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3355 Listeria...

  8. 21 CFR 866.3300 - Haemophilus spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Haemophilus spp. serological reagents. 866.3300 Section 866.3300 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3300...

  9. 21 CFR 866.3270 - Flavobacterium spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Flavobacterium spp. serological reagents. 866.3270 Section 866.3270 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents §...

  10. 21 CFR 866.3415 - Pseudomonas spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Pseudomonas spp. serological reagents. 866.3415 Section 866.3415 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3415...

  11. 21 CFR 866.3040 - Aspergillus spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Aspergillus spp. serological reagents. 866.3040 Section 866.3040 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3040...

  12. 21 CFR 866.3350 - Leptospira spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Leptospira spp. serological reagents. 866.3350 Section 866.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3350...

  13. 21 CFR 866.3340 - Klebsiella spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Klebsiella spp. serological reagents. 866.3340 Section 866.3340 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3340...

  14. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  15. 21 CFR 866.3125 - Citrobacter spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Citrobacter spp. serological reagents. 866.3125 Section 866.3125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3125...

  16. 21 CFR 866.3200 - Echinococcus spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Echinococcus spp. serological reagents. 866.3200 Section 866.3200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3200...

  17. Evaluation of a Non-Flowering Perennial Sorghum spp. Hybrid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Perennial Sorghum spp. hybrids (PSSHs) such as Columbusgrass (Sorghum almum Parodi; S. bicolor [L.] Moench x S. halepense [L.] Pers.) and the reciprocal hybridization (S. halepense x S. bicolor; e.g. Cv 'Krish') are high-biomass feedstocks currently utilized as forage but with potential as dual-...

  18. A culture method for darkling beetles, Blapstinus spp. (Coleoptera: Tenebrionidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Darkling beetles, Blapstinus spp., have become a serious pest of Cucurbitaceae crops, especially in California. A culture method was sought to provide large numbers (> 500) of adult beetles of known age and sex that could be used for laboratory testing when needed. A method previously developed for ...

  19. Asymptomatic presence of Nosema spp. in Spanish commercial apiaries.

    PubMed

    Fernández, José Manuel; Puerta, Francisco; Cousinou, Mercedes; Dios-Palomares, Rafaela; Campano, Francisco; Redondo, Laura

    2012-10-01

    Nosemosis is caused by intracellular parasites (Nosema apis and Nosema ceranae) that infect the midgut epithelial cells in adult honey bees. Recent studies relate N. ceranae to Colony Collapse Disorder and there is some suggestion that Nosema spp., especially N. ceranae, induces high mortality in honey bees, a fact that is considered as a serious threat for colony survival. 604 samples of adult honey bees for Nosema spp. analysis were collected from beekeeping colonies across Spain and were analysed using PCR with capillary electrophoresis. We also monitored 77 Andalusian apiaries for 2 years; the sampled hives were standard healthy colonies, without any special disease symptoms. We found 100% presence of Nosema spp. in some locations, indicating that this parasite was widespread throughout the country. The two year monitoring indicated that 87% of the hives with Nosema spp. remained viable, with normal honey production and biological development during this period of time. The results of these trials indicated that both N. ceranae and N. apis could be present in these beehives without causing disease symptom and that there is no evidence for the replacement of N. apis by N. ceranae, supporting the hypothesis that nosemosis is not the main reason of the collapse and death of beehives.

  20. Genetic diversity assessment of Musa spp. germplasm using SSR markers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The USDA-ARS Tropical Agriculture Research Station is responsible for conserving germplasm of a number of important agricultural crop species. Among these, a Musa spp. collection has been established and is comprised of diploid, triploid and tetraploid accessions of cultivated, ornamental, wild and...

  1. Adverse effects of larkspur (Delphinium spp.) on cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    There are numerous species of larkspurs (Delphinium spp.) in North America. The larkspurs are a major cause of cattle losses on western ranges in the USA, especially on foothill and mountain rangelands. The toxicity of larkspur species is due to various norditerpenoid alkaloids. In this article, we ...

  2. Lantana camara leaf extract mediated silver nanoparticles: Antibacterial, green catalyst.

    PubMed

    Ajitha, B; Ashok Kumar Reddy, Y; Shameer, Syed; Rajesh, K M; Suneetha, Y; Sreedhara Reddy, P

    2015-08-01

    Silver nanoparticles (AgNPs) have been synthesized by Lantana camara leaf extract through simple green route and evaluated their antibacterial and catalytic activities. The leaf extract (LE) itself acts as both reducing and stabilizing agent at once for desired nanoparticle synthesis. The colorless reaction mixture turns to yellowish brown attesting the AgNPs formation and displayed UV-Vis absorption spectra. Structural analysis confirms the crystalline nature and formation of fcc structured metallic silver with majority (111) facets. Morphological studies elicit the formation of almost spherical shaped nanoparticles and as AgNO3 concentration is increased, there is an increment in the particle size. The FTIR analysis evidences the presence of various functional groups of biomolecules of LE is responsible for stabilization of AgNPs. Zeta potential measurement attests the higher stability of synthesized AgNPs. The synthesized AgNPs exhibited good antibacterial activity when tested against Escherichia coli, Pseudomonas spp., Bacillus spp. and Staphylococcus spp. using standard Kirby-Bauer disc diffusion assay. Furthermore, they showed good catalytic activity on the reduction of methylene blue by L. camara extract which is monitored and confirmed by the UV-Vis spectrophotometer.

  3. 21 CFR 866.3720 - Streptococcus spp. exo-enzyme reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Streptococcus spp. exo-enzyme reagents. 866.3720 Section 866.3720 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... spp. exo-enzyme reagents. (a) Identification. Streptococcus spp. exoenzyme reagents are devices...

  4. 21 CFR 866.3720 - Streptococcus spp. exo-enzyme reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Streptococcus spp. exo-enzyme reagents. 866.3720 Section 866.3720 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... spp. exo-enzyme reagents. (a) Identification. Streptococcus spp. exoenzyme reagents are devices...

  5. 21 CFR 866.3720 - Streptococcus spp. exo-enzyme reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Streptococcus spp. exo-enzyme reagents. 866.3720 Section 866.3720 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... spp. exo-enzyme reagents. (a) Identification. Streptococcus spp. exoenzyme reagents are devices...

  6. Nisin resistance distinguishes Mycoplasma spp. from Acholeplasma spp. and provides a basis for selective growth media.

    PubMed Central

    Abu-Amero, K K; Halablab, M A; Miles, R J

    1996-01-01

    The sensitivity of 11 Mycoplasma and 5 Acholeplasma species to the bacteriocin nisin was determined. When applied on filter paper discs to lawns of acholeplasma cells, nisin (20 nmol per disc) gave 3.5- to 7.0-mm zones of growth inhibition. The inclusion of 0.2 mM nisin in agar medium reduced the number of Acholeplasma laidlawii colonies by a factor of more than 10(6), and in a salts solution, 75 microM nisin killed more than 99.9% of cells within 1 min. Under similar conditions, nisin had no significant effect upon the growth or survival of Mycoplasma species. At low concentrations (1 to 3 microM), nisin stimulated glucose oxidation by A. laidlawii and Acholeplasma oculi. However, in comparison with carbonyl cyanide m-chlorophenylhydrazone (CCCP), a recognized protonophore and uncoupler of respiration, the maximum extent of stimulation was low, < or = 20%, compared with up to 180% for CCCP. Also, in contrast to results obtained with CCCP, at concentrations only slightly above those causing stimulation of acholeplasma oxygen uptake, nisin strongly inhibited respiration. Inhibition of oxygen uptake was greater for A. laidlawii cells grown in the absence of cholesterol, and on agar medium, growth inhibition by nisin decreased with increasing concentrations of cholesterol. Nisin resistance may be a valuable characteristic in the selection and identification of Mycoplasma spp. PMID:11783455

  7. Enterotoxigenic Bacillus spp. DNA fingerprint revealed in naturally contaminated nonfat dry milk powder using rep-PCR.

    PubMed

    Cooper, Robin M; McKillip, John L

    2006-01-01

    Dry milk powders and functional ingredients frequently contain high levels of viable bacterial spores, some of which may result in growth of toxigenic Bacillus spp. in reconstituted and temperature-abused foods. Samples from nonfat dry milk (NFDM), infant milk formula (IMF), coffee creamer, lecithin, and cocoa powder were subjected to a short heat treatment followed by enrichment in tryptone phosphate glucose yeast extract (TPGY) broth at 32 degrees C for 12-25 hours to obtain cell densities of 10(6) CFU ml(-1). DNA was extracted using a modification of established protocol, leading to the development of an optimized method for each food system. Purified DNA was amplified by rep-PCR using extragenic sequence-targeting primers and optimized for each food. PCR fingerprints from each food were analyzed electrophoretically for banding patterns earlier correlated to that of enterotoxigenic Bacillus spp. and Bacillus cereus positive control DNA fingerprints. Reverse passive latex agglutination (RPLA) and Bacillus Diarrhoeal Enterotoxin Enzyme Linked Immunosorbent Assay (Tecra Diagnostics) confirmed the presence of HBL and NHE enterotoxin production in NFDM, Coffee creamer, infant milk formula, and two lecithin samples but not in cocoa powder. These results demonstrate the utility of rep-PCR not only as a tool for bacterial genotyping, but a unique means of quality control and hygiene monitoring in food microbiology.

  8. Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil.

    PubMed

    Rotondano, Tereza Emmanuelle de Farias; Almeida, Herta Karyanne Araújo; Krawczak, Felipe da Silva; Santana, Vanessa Lira; Vidal, Ivana Fernandes; Labruna, Marcelo Bahia; de Azevedo, Sérgio Santos; Ade lmeida, Alzira Maria Paiva; de Melo, Marcia Almeida

    2015-01-01

    This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100) of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR), respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100) of the animals. Babesia vogeli was detected by PCR in ten animals (10%) and was correlated with young age (p = 0.007) and thrombocytopenia (p = 0.01). None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State. PMID:25909253

  9. Detection of Salmonella spp., Candida albicans, Aspergillus spp., and Antimicrobial Residues in Raw and Processed Cow Milk from Selected Smallholder Farms of Zimbabwe

    PubMed Central

    Mhone, Tryness Anastazia; Matope, Gift; Saidi, Petronella Tapiwa

    2012-01-01

    A cross-sectional study was conducted to detect the presence of Salmonella spp., Candida albicans, Aspergillus spp., and antimicrobial residues in raw milk (n = 120) and processed cow milk (n = 20) from smallholder dairy farms from three sites in Zimbabwe. Culture and isolation of Salmonella spp., C. albicans, and Aspergillus spp. were performed using selective media, while antimicrobial residues were detected by a dye reduction test. No Salmonella, but C. albicans (17.5%; 21/120), Aspergillus spp. (0.8%; 1/120), and antimicrobial residues (2.5%; 3/120) were detected from raw milk. C. albicans was isolated from all three sites, while Aspergillus spp. and antimicrobial residues were detected from sites 1 and 3, respectively. From processed milk, only C. albicans (5%) was isolated while Aspergillus spp. and antimicrobial residues were not detected. These results suggested low prevalence of Salmonella spp. and Aspergillus spp. and a relatively high prevalence of C. albicans in raw milk from the smallholder farms. The potential public health risks of C. albicans and the detected antimicrobial residues need to be considered. Thus, educating farmers on improving milking hygiene and storage of milk and establishing programmes for monitoring antimicrobial residues may help to improve the safety of milk from smallholder farms. PMID:23050199

  10. Occurrence of Cladosporium spp. and Alternaria spp. spores in Western, Northern and Central-Eastern Poland in 2004-2006 and relation to some meteorological factors

    NASA Astrophysics Data System (ADS)

    Grinn-Gofroń, Agnieszka; Rapiejko, Piotr

    2009-08-01

    The concentration of airborne spores of Cladosporium spp. and Alternaria spp. has been investigated at three monitoring stations situated along the west-north and central-east transect in Poland (Szczecin, Olsztyn, Warszawa,) i.e. from a height of 100 m to 149 m above sea level. The aerobiological monitoring of fungal spores was performed by means of three Lanzoni volumetric spore traps. Cladosporium spp. spores were dominant at all the stations. The highest Cladosporium spp. and Alternaria spp. numbers of spores were observed at all the cities in July and August. Statistically significant correlations have been found between the Cladosporium spp. and Alternaria spp. concentration in the air and the mean air temperature, amount of precipitation, air pressure and relative air humidity. The spore count of Cladosporium spp. and Alternaria spp. is determined by the diversity of local flora and weather conditions, especially by the air temperature. The identification of factors, which influence and shape spore concentrations, may significantly improve the current methods of allergy prevention.

  11. Screening of allelopathic trees for their antifungal potential against Alternaria alternata strains isolated from dying-back Eucalyptus spp.

    PubMed

    Javaid, Arshad; Samad, Sara

    2012-01-01

    Antifungal activity of methanolic extracts of leaves of three tree species, namely Azadirachta indica L., Syzygium cumini (L.) Skeels. and Melia azedarach L. was evaluated against two strains of Alternaria alternata, isolated from dying-back trees of two Eucalyptus spp., namely Eucalyptus citriodora and Eucalyptus globulus. All the concentrations (1, 2, … , 5% w/v) of the methanolic extracts of the three tree species significantly reduced the fungal biomass. There were reductions in the ranges 82-88%, 88-96% and 83-96% in the biomass of A. alternata strains due to different concentrations of the leaf extracts of S. cumini, A. indica and M. azedarach, respectively. Methanolic extract of M. azedarach was subjected to further fractionation using n-hexane, chloroform, ethyl acetate and n-butanol successively in the order of increasing polarity. Aqueous and n-butanol fractions gave promising results in the significant decrease in fungal biomass. This study concludes that aqueous and n-butanol fractions of methanolic leaf extract of M. azedarach can be used as biofungicides for the management of A. alternata. PMID:22007991

  12. First reported case of turtle deaths during a toxic Microcystis spp. bloom in Lake Oubeira, Algeria.

    PubMed

    Nasri, Hichem; El Herry, Soumaya; Bouaïcha, Noureddine

    2008-10-01

    Microcystins analysis was conducted in field cyanobacterial bloom samples and dead terrapin tissues from Lake Oubeira (Algeria) with an aim of studying the cause of the mortality of the freshwater terrapin species Emys orbicularis and Mauremys leprosa during October 2005. The deaths of these two terrapin species were observed during a bloom of Microcystis spp. The total microcystin content per phytoplankton biomass evaluated with the methanol extraction-protein phosphatase methodology was 1.12 mg MCYST-LR equivalents/g dried bloom material. The analysis of this bloom extract by the LC/MS technique demonstrated the presence of three microcystin variants: microcystin-LR (MCYST-LR), microcystin-YR (MCYST-YR), and microcystin-RR (MCYST-RR). Microcystins were also detected in fresh carcasses of terrapin liver, viscera and muscle tissues using the GC/MS after Lemieux oxidation method and the PP2A inhibition assay. The high level of microcystins detected using the Lemieux oxidation-GC/MS method in the liver tissue (1192.8 microg MCYST-LR equivalent/g dw) and in the viscera tissue (37.19 microg MCYST-LR equivalent/g dw) of the species M. leprosa and E. orbicularis, respectively, and the liver crumbling observed after the necropsy examination of the fresh carcass of M. leprosa support the possibility that cyanobacterial microcystins contribute to the turtle mortalities. PMID:18234335

  13. Phenolics, antioxidant capacity and bioaccessibility of chicory varieties (Cichorium spp.) grown in Turkey.

    PubMed

    Sahan, Yasemin; Gurbuz, Ozan; Guldas, Metin; Degirmencioglu, Nurcan; Begenirbas, Aynur

    2017-02-15

    In this study, the changes in phenolics, anthocyanin, antioxidant capacity, and bioaccessibility of chicory varieties (Cichorium spp.) in Turkey were investigated. A total of 19 phenolic standards were screened in the chicory varieties studied and the most abundant compounds in the samples, extracted with methanol, were phenolic acids, syringic (2.54mg/kg) and trans-ferulic acid (1.85mg/kg), whilst (+)-catechin was the major flavanol. The highest flavanol content using either methanol or ethanol was determined in the green chicory samples (0.62mg/kg). The red chicory variety had higher anthocyanin (12.80mg/kg), and contained more phenolics, extractable (8855.50mg GAE/100g) and hydrolysable (7005.51mg GAE/100g), than the other varieties. Also, the antioxidant capacities in this variety, as measured using the CUPRAC assay (570.54 and 425.14μmol Trolox/g dw, respectively), had a wider range of difference than was found in the other assays used. Total phenolics were more bioaccessible from the white chicory variety (61.48%). However, the bioaccessibility of antioxidants was higher in the green chicory variety. PMID:27664662

  14. A comparison of a new centrifuge sugar flotation technique with the agar method for the extraction of immature Culicoides (Diptera: Ceratopogonidae) life stages from salt marsh soils.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two sampling techniques, agar extraction (AE) and centrifuge sugar flotation extraction (CSFE) were compared to determine their relative efficacy to recover immature stages of Culicoides spp from salt marsh substrates. Three types of samples (seeded with known numbers of larvae, homogenized field s...

  15. Morphology and mycelial growth rate of Pleurotus spp. strains from the Mexican mixtec region.

    PubMed

    Guadarrama-Mendoza, P C; del Toro, G Valencia; Ramírez-Carrillo, R; Robles-Martínez, F; Yáñez-Fernández, J; Garín-Aguilar, M E; Hernández, C G; Bravo-Villa, G

    2014-01-01

    Two native Pleurotus spp. strains (white LB-050 and pale pink LB-051) were isolated from rotten tree trunks of cazahuate (Ipomoea murucoides) from the Mexican Mixtec Region. Both strains were chemically dedikaryotized to obtain their symmetrical monokaryotic components (neohaplonts). This was achieved employing homogenization time periods from 60 to 65 s, and 3 day incubation at 28 °C in a peptone-glucose solution (PGS). Pairing of compatible neohaplonts resulted in 56 hybrid strains which were classified into the four following hybrid types: (R(1-n)xB(1-n), R(1-n)xB(2-1), R(2-n)xB(1-n) and R(2-n)xB(2-1)). The mycelial growth of Pleurotus spp. monokaryotic and dikaryotic strains showed differences in texture (cottony or floccose), growth (scarce, regular or abundant), density (high, regular or low), and pigmentation (off-white, white or pale pink). To determine the rate and the amount of mycelium growth in malt extract agar at 28 °C, the diameter of the colony was measured every 24 h until the Petri dish was completely colonized. A linear model had the best fit to the mycelial growth kinetics. A direct relationship between mycelial morphology and growth rate was observed. Cottony mycelium presented significantly higher growth rates (p < 0.01) in comparison with floccose mycelium. Thus, mycelial morphology can be used as criterion to select which pairs must be used for optimizing compatible-mating studies. Hybrids resulting from cottony neohaplonts maintained the characteristically high growth rates of their parental strains with the hybrid R(1-n)xB(1-n) being faster than the latter. PMID:25477920

  16. Stable-isotope probing implicates Methylophaga spp and novel Gammaproteobacteria in marine methanol and methylamine metabolism.

    PubMed

    Neufeld, Josh D; Schäfer, Hendrik; Cox, Michael J; Boden, Rich; McDonald, Ian R; Murrell, J Colin

    2007-10-01

    The metabolism of one-carbon (C(1)) compounds in the marine environment affects global warming, seawater ecology and atmospheric chemistry. Despite their global significance, marine microorganisms that consume C(1) compounds in situ remain poorly characterized. Stable-isotope probing (SIP) is an ideal tool for linking the function and phylogeny of methylotrophic organisms by the metabolism and incorporation of stable-isotope-labelled substrates into nucleic acids. By combining DNA-SIP and time-series sampling, we characterized the organisms involved in the assimilation of methanol and methylamine in coastal sea water (Plymouth, UK). Labelled nucleic acids were analysed by denaturing gradient gel electrophoresis (DGGE) and clone libraries of 16S rRNA genes. In addition, we characterized the functional gene complement of labelled nucleic acids with an improved primer set targeting methanol dehydrogenase (mxaF) and newly designed primers for methylamine dehydrogenase (mauA). Predominant DGGE phylotypes, 16S rRNA, methanol and methylamine dehydrogenase gene sequences, and cultured isolates all implicated Methylophaga spp, moderately halophilic marine methylotrophs, in the consumption of both methanol and methylamine. Additionally, an mxaF sequence obtained from DNA extracted from sea water clustered with those detected in (13)C-DNA, suggesting a predominance of Methylophaga spp among marine methylotrophs. Unexpectedly, most predominant 16S rRNA and functional gene sequences from (13)C-DNA were clustered in distinct substrate-specific clades, with 16S rRNA genes clustering with sequences from the Gammaproteobacteria. These clades have no cultured representatives and reveal an ecological adaptation of particular uncultured methylotrophs to specific C(1) compounds in the coastal marine environment.

  17. Acanthamoeba spp. in Contact Lenses from Healthy Individuals from Madrid, Spain

    PubMed Central

    Gomes, Thiago dos Santos; Magnet, Angela; Izquierdo, Fernando; Vaccaro, Lucianna; Redondo, Fernando; Bueno, Sara; Sánchez, Maria Luisa; Angulo, Santiago; Fenoy, Soledad; Hurtado, Carolina; del Aguila, Carmen

    2016-01-01

    Purpose Acanthamoeba keratitis (AK) is a painful and potentially blinding corneal infection caused by Acanthamoeba spp. In Madrid, environmental studies have demonstrated a high presence of these free-living amoebae in tap water. Since most of AK cases occur in contact lenses (CL) wearers with inadequate hygiene habits, the presence of Acanthamoeba in discarded CL has been studied and compared with other common etiological agents of keratitis, such as Pseudomonas aeruginosa and Staphylococcus aureus. Methods One hundred and seventy-seven healthy individuals from Madrid contributed their discarded CL and answered a questionnaire on hygiene habits. DNA was extracted from the CL solution and analyzed by real-time PCR for Acanthamoeba, Pseudomonas aeruginosa and Staphylococcus aureus. These CL and their solutions were also cultured on non-nutrient agar to isolate Acanthamoeba. Results Among the 177 samples, Acanthamoeba DNA was detected in 87 (49.2%), P. aeruginosa DNA in 14 (7.9%) and S. aureus DNA in 19 (10.7%). Cultivable amoebae, however, were observed in only one sample (0.6%). This isolate was genotyped as T4. The habits reported by this CL owner included some recognized risk factors for AK, but in this study only the practice of “not cleaning the CL case” presented some statistical significant association with Acanthamoeba DNA presence. Detection of the investigated bacterial DNA did not demonstrate statistical significant association with the studied practices, but the presence of P. aeruginosa revealed a possible inhibition of Acanthamoeba in these samples. Conclusions The PCR results suggest a high presence of Acanthamoeba spp. in healthy CL wearers from Madrid, but we can assume that CL solutions are properly disinfecting the CL since only 1.1% of the positive PCR samples correspond to viable amoebae and, after four years, only one participant reported stronger ocular problems. Nevertheless, more studies are necessary to corroborate this hypothesis. PMID

  18. Morphology and mycelial growth rate of Pleurotus spp. strains from the Mexican mixtec region.

    PubMed

    Guadarrama-Mendoza, P C; del Toro, G Valencia; Ramírez-Carrillo, R; Robles-Martínez, F; Yáñez-Fernández, J; Garín-Aguilar, M E; Hernández, C G; Bravo-Villa, G

    2014-01-01

    Two native Pleurotus spp. strains (white LB-050 and pale pink LB-051) were isolated from rotten tree trunks of cazahuate (Ipomoea murucoides) from the Mexican Mixtec Region. Both strains were chemically dedikaryotized to obtain their symmetrical monokaryotic components (neohaplonts). This was achieved employing homogenization time periods from 60 to 65 s, and 3 day incubation at 28 °C in a peptone-glucose solution (PGS). Pairing of compatible neohaplonts resulted in 56 hybrid strains which were classified into the four following hybrid types: (R(1-n)xB(1-n), R(1-n)xB(2-1), R(2-n)xB(1-n) and R(2-n)xB(2-1)). The mycelial growth of Pleurotus spp. monokaryotic and dikaryotic strains showed differences in texture (cottony or floccose), growth (scarce, regular or abundant), density (high, regular or low), and pigmentation (off-white, white or pale pink). To determine the rate and the amount of mycelium growth in malt extract agar at 28 °C, the diameter of the colony was measured every 24 h until the Petri dish was completely colonized. A linear model had the best fit to the mycelial growth kinetics. A direct relationship between mycelial morphology and growth rate was observed. Cottony mycelium presented significantly higher growth rates (p < 0.01) in comparison with floccose mycelium. Thus, mycelial morphology can be used as criterion to select which pairs must be used for optimizing compatible-mating studies. Hybrids resulting from cottony neohaplonts maintained the characteristically high growth rates of their parental strains with the hybrid R(1-n)xB(1-n) being faster than the latter.

  19. The outer membranes of Brucella spp. are resistant to bactericidal cationic peptides.

    PubMed Central

    Martínez de Tejada, G; Pizarro-Cerdá, J; Moreno, E; Moriyón, I

    1995-01-01

    The actions of polymyxin B, rabbit polymorphonuclear lysosome extracts, 14 polycationic peptides (including defensin NP-2, cecropin P1, lactoferricin B, and active peptides from cationic protein 18 and bactenecin), EDTA, and Tris on Brucella spp. were studied, with other gram-negative bacteria as controls. Brucella spp. were comparatively resistant to all of the agents listed above and bound less polymyxin B, and their outer membranes (OMs) were neither morphologically altered nor permeabilized to lysozyme by polymyxin B concentrations, although both effects were observed for controls. EDTA and peptides increased or accelerated the partition of the hydrophobic probe N-phenyl-naphthylamine into Escherichia coli and Haemophilus influenzae OMs but had no effect on Brucella OMs. Since Brucella and H. influenzae OMs are permeable to hydrophobic compounds (G. Martínez de Tejada and I. Moriyón, J. Bacteriol. 175:5273-5275, 1993), the results show that such unusual permeability is not necessarily related to resistance to polycations. Although rough (R) B. abortus and B. ovis were more resistant than the controls were, there were qualitative and quantitative differences with smooth (S) brucellae; this may explain known host range and virulence differences. Brucella S-lipopolysaccharides (LPSs) had reduced affinities for polycations, and insertion of Brucella and Salmonella montevideo S-LPSs into the OM of a Brucella R-LPS mutant increased and decreased, respectively, its resistance to cationic peptides. The results show that the core lipid A of Brucella LPS plays a major role in polycation resistance and that O-chain density also contributes significantly. It is proposed that the features described above contribute to Brucella resistance to the oxygen-independent systems of phagocytes. PMID:7622230

  20. Morphology and mycelial growth rate of Pleurotus spp. strains from the Mexican mixtec region

    PubMed Central

    Guadarrama-Mendoza, P.C.; del Toro, G. Valencia; Ramírez-Carrillo, R.; Robles-Martínez, F.; Yáñez-Fernández, J.; Garín-Aguilar, M.E.; Hernández, C.G.; Bravo-Villa, G.

    2014-01-01

    Two native Pleurotus spp. strains (white LB-050 and pale pink LB-051) were isolated from rotten tree trunks of cazahuate (Ipomoea murucoides) from the Mexican Mixtec Region. Both strains were chemically dedikaryotized to obtain their symmetrical monokaryotic components (neohaplonts). This was achieved employing homogenization time periods from 60 to 65 s, and 3 day incubation at 28 °C in a peptone-glucose solution (PGS). Pairing of compatible neohaplonts resulted in 56 hybrid strains which were classified into the four following hybrid types: (R1-nxB1-n, R1-nxB2-1, R2-nxB1-n and R2-nxB2-1). The mycelial growth of Pleurotus spp. monokaryotic and dikaryotic strains showed differences in texture (cottony or floccose), growth (scarce, regular or abundant), density (high, regular or low), and pigmentation (off-white, white or pale pink). To determine the rate and the amount of mycelium growth in malt extract agar at 28 °C, the diameter of the colony was measured every 24 h until the Petri dish was completely colonized. A linear model had the best fit to the mycelial growth kinetics. A direct relationship between mycelial morphology and growth rate was observed. Cottony mycelium presented significantly higher growth rates (p < 0.01) in comparison with floccose mycelium. Thus, mycelial morphology can be used as criterion to select which pairs must be used for optimizing compatible-mating studies. Hybrids resulting from cottony neohaplonts maintained the characteristically high growth rates of their parental strains with the hybrid R1-nxB1-n being faster than the latter. PMID:25477920

  1. Characterization and virulence of Beauveria spp. recovered from emerald ash borer in southwestern Ontario, Canada.

    PubMed

    Johny, Shajahan; Kyei-Poku, George; Gauthier, Debbie; Frankenhuyzen, Kees van; Krell, Peter J

    2012-09-15

    The emerald ash borer (EAB), Agrilus planipennis (Coleoptera: Buprestidae), is an invasive wood boring beetle that is decimating North America's ash trees (Fraxinus spp.). To find effective and safe indigenous biocontrol agents to manage EAB, we conducted a survey in 2008-2009 of entomopathogenic fungi (EPF) infecting EAB in five outbreak sites in southwestern Ontario, Canada. A total of 78 Beauveria spp. isolates were retrieved from dead and mycosed EAB cadavers residing in the phloem tissues of dead ash barks, larval frass extracted from feeding galleries under the bark of dead trees. Molecular characterization using sequences of the ITS, 5' end of EF1-α and intergenic Bloc region fragments revealed that Beauveria bassiana and Beauveria pseudobassiana were commonly associated with EAB in the sampled sites. Based on phylogenetic analysis inferred from ITS sequences, 17 of these isolates clustered with B. bassiana, which further grouped into three different sub-clades. However, the combined EF1-α and Bloc sequences detected five genotypes among the three sub-clades. The remaining 61 isolates clustered with B. pseudobassiana, which had identical ITS sequences but were further subdivided into two genotypes by variation in the EF1-α and Bloc regions. Initial virulence screening against EAB adults of 23 isolates representing the different clades yielded 8 that produced more than 90% mortality in a single concentration assay. These isolates differed in virulence based on LC(50) values estimated from multiple concentration bioassay and based on mean survival times at a conidia concentration of 2×10(6) conidia/ml. B. bassiana isolate L49-1AA was significantly more virulent and produced more conidia on EAB cadavers compared to the other indigenous isolates and the commercial strain B. bassiana GHA suggesting that L49-1AA may have potential as a microbiological control agent against EAB.

  2. Detection of Ehrlichia spp., Anaplasma spp., Rickettsia spp., and other eubacteria in ticks from the Thai-Myanmar border and Vietnam.

    PubMed

    Parola, Philippe; Cornet, Jean-Paul; Sanogo, Yibayiri Osée; Miller, R Scott; Thien, Huynh Van; Gonzalez, Jean-Paul; Raoult, Didier; Telford III, Sam R; Wongsrichanalai, Chansuda

    2003-04-01

    A total of 650 ticks, including 13 species from five genera, were collected from animals, from people, or by flagging of the vegetation at sites on the Thai-Myanmar border and in Vietnam. They were tested by PCR to detect DNA of bacteria of the order RICKETTSIALES: Three Anaplasma spp. were detected in ticks collected in Thailand, including (i) Anaplasma sp. strain AnDa465, which was considered a genotype of Anaplasma platys (formerly Ehrlichia platys) and which was obtained from Dermacentor auratus ticks collected from dogs; (ii) Anaplasma sp. strain AnAj360, which was obtained from Amblyomma javanense ticks collected on a pangolin; and (iii) Anaplasma sp. strain AnHl446, which was closely related to Anaplasma bovis and which was detected in Haemaphysalis lagrangei ticks collected from a bear. Three Ehrlichia spp. were identified, including (i) Ehrlichia sp. strain EBm52, which was obtained from Boophilus microplus ticks collected from cattle from Thailand; (ii) Ehrlichia sp. strain EHh324, which was closely related to Ehrlichia chaffeensis and which was detected in Haemaphysalis hystricis ticks collected from wild pigs in Vietnam; and (iii) Ehrlichia sp. strain EHh317, which was closely related to Ehrlichia sp. strain EBm52 and which was also detected in H. hystricis ticks collected from wild pigs in Vietnam. Two Rickettsia spp. were detected in Thailand, including (i) Rickettsia sp. strain RDla420, which was detected in Dermacentor auratus ticks collected from a bear, and (ii) Rickettsia sp. strain RDla440, which was identified from two pools of Dermacentor larvae collected from a wild pig nest. Finally, two bacteria named Eubacterium sp. strain Hw124 and Eubacterium sp. strain Hw191 were identified in Haemaphysalis wellingtoni ticks collected from chicken in Thailand; these strains could belong to a new group of bacteria. PMID:12682151

  3. Genetic variation in codons 167, 198 and 200 of the beta-tubulin gene in whipworms (Trichuris spp.) from a range of domestic animals and wildlife.

    PubMed

    Hansen, Tina V A; Nejsum, Peter; Olsen, Annette; Thamsborg, Stig Milan

    2013-03-31

    A recurrent problem in the control of whipworm (Trichuris spp.) infections in many animal species and man is the relatively low efficacy of treatment with a single application of benzimidazoles (BZs). The presence of single nucleotide polymorphisms (SNPs) in codons 167, 198 and 200 in the beta-tubulin gene has been associated with BZ anthelmintic resistance in intestinal nematodes of veterinary importance. We hypothesized that the low susceptibility to BZ could be related to a natural tolerance or induced resistance caused by BZ-resistant associated SNPs. The aim of the present study was therefore to investigate the presence of these SNPs in the beta-tubulin gene of Trichuris spp. obtained from a range of animals. DNA was extracted from a total of 121 Trichuris spp. adult whipworm specimens obtained from 6 different host species. The number of worms from each host was pig: 31, deer: 21, sheep: 18, mouse: 17, dog: 19 and Arabian camels: 14. A pooled sample of Trichuris eggs from 3 moose was also used. In order to amplify the beta-tubulin fragments which covered codons 167, 198 and 200 of the gene, degenerate primers were designed. The sequences obtained were used to design species specific primers and used to amplify a ~476 bp fragment of the beta-tubulin gene. The PCR products were sequenced, analysed and evaluated. We did not identify SNPs in codons 167, 198 or 200 that led to amino acid substitutions in any of the studied Trichuris spp., but genetic variation expected to be related to species differences was observed. The cluster analysis showed close evolutionary relationship between Trichuris spp. from ruminants and between mouse and dog whereas the pig-derived worms, T. suis, clustered with T. trichiura obtained from Genbank.

  4. Bloodstream infections due to Peptoniphilus spp.: report of 15 cases

    PubMed Central

    Brown, K; Church, D; Lynch, T; Gregson, D

    2014-01-01

    Peptoniphilus spp. are Gram-positive anaerobic cocci (GPAC) that were formerly classified in the genus Peptostreptococcus. This study describes 15 cases of Peptoniphilus spp. bloodstream infection (BSI) diagnosed from 2007 to 2011 using 16S rDNA sequencing in patients with pneumonia, pre-term delivery, soft tissue infection or colon or bladder disease. Seven out of 15 (47%) of these cases had polymicrobial BSIs. One of the isolates was closely related to P. duerdenii (EU526290), while the other 14 isolates were most closely related to a Peptoniphilus sp. reference strain (ATCC 29743) and P. hareii (Y07839). Peptoniphilus is a rare but important cause of BSI. PMID:24773457

  5. Incidence of Listeria spp. in vegetables in Kuala Lumpur.

    PubMed

    Tang, M Y; Cheong, Y M; Zainuldin, T

    1994-09-01

    From April 1992 to September 1992, 280 samples of 10 different fresh vegetables, bought from four different market outlets in Kuala Lumpur were examined for the presence of Listeria spp. Most of the market produce were locally grown with the exception of carrots. The isolation procedure was based on the Food & Drug Administration method (modified) used for the detection of Listeria spp. Isolation media used were Listeria Selective medium and LiCl- phenylethanol-Moxalactam agars. The identification of isolates was by means of conventional biochemical tests and API Listeria identification system. Five out of the 280 samples showed Listeria contamination, Listeria monocytogenes was isolated in lettuce, sengkuang (Pachyrrhizus erosus) and selom Oenanthe javanica) and Listeria innocua was isolated from sengkuang (Pachyrrhizus erosus) and pegaga (Hydrocotyle asiatica).

  6. First Isolates of Leptospira spp., from Rodents Captured in Angola

    PubMed Central

    Fortes-Gabriel, Elsa; Carreira, Teresa; Vieira, Maria Luísa

    2016-01-01

    Rodents play an important role in the transmission of pathogenic Leptospira spp. However, in Angola, neither the natural reservoirs of these spirochetes nor leptospirosis diagnosis has been considered. Regarding this gap, we captured rodents in Luanda and Huambo provinces to identify circulating Leptospira spp. Rodent kidney tissue was cultured and DNA amplified and sequenced. Culture isolates were evaluated for pathogenic status and typing with rabbit antisera; polymerase chain reaction (PCR) and sequencing were also performed. A total of 37 rodents were captured: Rattus rattus (15, 40.5%), Rattus norvegicus (9, 24.3%), and Mus musculus (13, 35.2%). Leptospiral DNA was amplified in eight (21.6%) kidney samples. From the cultures, we obtained four (10.8%) Leptospira isolates belonging to the Icterohaemorrhagiae and Ballum serogroups of Leptospira interrogans and Leptospira borgpetersenii genospecies, respectively. This study provides information about circulating leptospires spread by rats and mice in Angola. PMID:26928840

  7. First Isolates of Leptospira spp., from Rodents Captured in Angola.

    PubMed

    Fortes-Gabriel, Elsa; Carreira, Teresa; Vieira, Maria Luísa

    2016-05-01

    Rodents play an important role in the transmission of pathogenic Leptospira spp. However, in Angola, neither the natural reservoirs of these spirochetes nor leptospirosis diagnosis has been considered. Regarding this gap, we captured rodents in Luanda and Huambo provinces to identify circulating Leptospira spp. Rodent kidney tissue was cultured and DNA amplified and sequenced. Culture isolates were evaluated for pathogenic status and typing with rabbit antisera; polymerase chain reaction (PCR) and sequencing were also performed. A total of 37 rodents were captured: Rattus rattus (15, 40.5%), Rattus norvegicus (9, 24.3%), and Mus musculus (13, 35.2%). Leptospiral DNA was amplified in eight (21.6%) kidney samples. From the cultures, we obtained four (10.8%) Leptospira isolates belonging to the Icterohaemorrhagiae and Ballum serogroups of Leptospira interrogans and Leptospira borgpetersenii genospecies, respectively. This study provides information about circulating leptospires spread by rats and mice in Angola.

  8. SPP: A data base processor data communications protocol

    NASA Technical Reports Server (NTRS)

    Fishwick, P. A.

    1983-01-01

    The design and implementation of a data communications protocol for the Intel Data Base Processor (DBP) is defined. The protocol is termed SPP (Service Port Protocol) since it enables data transfer between the host computer and the DBP service port. The protocol implementation is extensible in that it is explicitly layered and the protocol functionality is hierarchically organized. Extensive trace and performance capabilities have been supplied with the protocol software to permit optional efficient monitoring of the data transfer between the host and the Intel data base processor. Machine independence was considered to be an important attribute during the design and implementation of SPP. The protocol source is fully commented and is included in Appendix A of this report.

  9. Sarcocystis spp. Infection in two Red Panda Cubs (Ailurus fulgens).

    PubMed

    Zoll, W M; Needle, D B; French, S J; Lim, A; Bolin, S; Langohr, I; Agnew, D

    2015-01-01

    Two neonatal male red panda (Ailurus fulgens) littermates were submitted for necropsy examination. One animal was found dead with no prior signs of illness; the other had a brief history of laboured breathing. Post-mortem examination revealed disseminated protozoal infection. To further characterize the causative agent, transmission electron microscopy (TEM), immunohistochemistry (IHC), polymerase chain reaction (PCR) and amplification and nucleic acid sequencing were performed. IHC was negative for Toxoplasma gondii and Neospora caninum, but was positive for a Sarcocystis spp. TEM of cardiac muscle and lung revealed numerous intracellular apicomplexan protozoa within parasitophorous vacuoles. PCR and nucleic acid sequencing of partial 18S rRNA and the internal transcribed spacer (ITS)-1 region confirmed a Sarcocystis spp. that shared 99% sequence homology to Sarcocystis neurona and Sarcocystis dasypi. This represents the first report of sarcocystosis in red pandas. The histopathological, immunohistochemical, molecular and ultrastructural findings are supportive of vertical transmission resulting in fatal disseminated disease.

  10. Yersinia spp. in surface water in Matsue, Japan.

    PubMed

    Fukushima, H; Saito, K; Tsubokura, M; Otsuki, K

    1984-06-01

    Yersinia spp. (741 strains) were recovered from 81% of 48 surface water samples collected over a 12-month period from four rivers in Matsue, Japan. The precipitation methods with FeCl3 or Kaolin and the cold enrichment method with Peptone-Mannitol-Phosphate buffer solution were used for recovery. Isolates belonged to Yersinia enterocolitica (Ye) (133 strains), Yersinia intermedia (511 strains), Yersinia frederiksenii (57 strains), Yersinia kristensenii (10 strains) and X2-like organisms (30 strains). Thirty colonies of Ye O3 biotype 3 per ml surface water may relate to the drainage containing 2 X 10(4) Ye O3 biotype 3 per ml, from the piggery that raised Ye O3 biotype 3-positive pigs. There was the negative interrelation between the incidence of isolation of Yersinia spp. and the environmental- and water temperatures. This may be the first documentation of isolation of Ye O3 from surface water.

  11. Sarcocystis spp. Infection in two Red Panda Cubs (Ailurus fulgens).

    PubMed

    Zoll, W M; Needle, D B; French, S J; Lim, A; Bolin, S; Langohr, I; Agnew, D

    2015-01-01

    Two neonatal male red panda (Ailurus fulgens) littermates were submitted for necropsy examination. One animal was found dead with no prior signs of illness; the other had a brief history of laboured breathing. Post-mortem examination revealed disseminated protozoal infection. To further characterize the causative agent, transmission electron microscopy (TEM), immunohistochemistry (IHC), polymerase chain reaction (PCR) and amplification and nucleic acid sequencing were performed. IHC was negative for Toxoplasma gondii and Neospora caninum, but was positive for a Sarcocystis spp. TEM of cardiac muscle and lung revealed numerous intracellular apicomplexan protozoa within parasitophorous vacuoles. PCR and nucleic acid sequencing of partial 18S rRNA and the internal transcribed spacer (ITS)-1 region confirmed a Sarcocystis spp. that shared 99% sequence homology to Sarcocystis neurona and Sarcocystis dasypi. This represents the first report of sarcocystosis in red pandas. The histopathological, immunohistochemical, molecular and ultrastructural findings are supportive of vertical transmission resulting in fatal disseminated disease. PMID:26054654

  12. Factors affecting Brucella spp. blood cultures positivity in children.

    PubMed

    Apa, Hurşit; Devrim, Ilker; Memur, Seyma; Günay, Ilker; Gülfidan, Gamze; Celegen, Mehmet; Bayram, Nuri; Karaarslan, Utku; Bağ, Ozlem; Işgüder, Rana; Oztürk, Aysel; Inan, Seyhan; Unal, Nurrettin

    2013-03-01

    Brucella infections have a wide spectrum of symptoms especially in children, making the diagnosis a complicated process. The gold standard for the final diagnosis for brucellosis is to identify the Brucella spp. isolated from blood or bone marrow cultures. The main purpose of this work was to evaluate the factors affecting the isolation of Brucella spp. from blood cultures. In our study, the ratio of fever, presence of hepatomegaly, and splenomegaly were found to be higher in the bacteremic group. In addition, C-reactive protein levels and liver function enzymes were found to be higher in the bacteremic group. In our opinion, while evaluating the febrile child with suspected Brucella infection, we highly recommend sampling blood cultures regardless of the history of previous antimicrobial therapy and duration of the symptoms.

  13. Efficacy of moxidectin against multiple resistant Ostertagia spp. in lambs.

    PubMed

    Várady, M; Praslicka, J; Corba, J

    1995-06-01

    Moxidectin was demonstrated to have a high efficacy in lambs against Ostertagia spp. which were resistant to albendazole, levamisole and ivermectin in goats. Moxidectin reduced the number of eggs in faeces by 99.6% and the number of worms found at post-mortem dissection of the lambs by 99.9%. Of the adult worms found in abomasa, 91% were identified as Ostertagia circumcincta and 9% as Ostertagia trifurcata.

  14. Presence of Cryptosporidium spp. and Giardia duodenalis through drinking water.

    PubMed

    Castro-Hermida, José Antonio; García-Presedo, Ignacio; Almeida, André; González-Warleta, Marta; Correia Da Costa, José Manuel; Mezo, Mercedes

    2008-11-01

    To evaluate the presence of Cryptosporidium spp. and Giardia duodenalis in the influent and final effluent of sixteen drinking water treatment plants located in a hydrographic basin in Galicia (NW Spain) - in which the principal river is recognised as a Site of Community Importance (SCI) - estimate the efficiency of treatment plants in removing these protozoans and determine the species and genotypes of the parasites by means of a molecular assay. All plant samples of influent and final effluent (50-100 l) were examined in the spring, summer, autumn and winter of 2007. A total of 128 samples were analysed by method 1623, developed by US Environmental Protection Agency for isolation and detection of both parasites. To identify the genotypes present the following genes were amplified and sequenced: 18S SSU rRNA (Cryptosporidium spp.) and b-giardina (G. duodenalis). The mean concentrations of parasites in the influent were 0.0-10.5 Cryptosporidium spp. oocysts per litre and 1.0-12.8 of G. duodenalis cysts per litre. In the final treated effluent, the mean concentration of parasites ranged from 0.0-3.0 oocysts per litre and 0.5-4.0 cysts per litre. The distribution of results by season revealed that in all plants, the highest numbers of (oo)cysts were recorded in spring and summer. Cryptosporidium parvum, C. andersoni, C. hominis and assemblages A-I, A-II, E of G. duodenalis were detected. Cryptosporidium spp. and G. duodenalis were consistently found at high concentrations in drinking water destined for human and animal consumption in the hydrographic basin under study, in Galicia (NW Spain). It is important that drinking water treatment authorities rethink the relevance of contamination levels of both parasites in drinking water and develop adequate countermeasures.

  15. Seroprevalence of enteropathogenic Yersinia spp. in pig batches at slaughter.

    PubMed

    Vanantwerpen, Gerty; Van Damme, Inge; De Zutter, Lieven; Houf, Kurt

    2014-09-01

    Enteropathogenic Yersinia spp. are one of the main causes of foodborne bacterial infections in Europe. Slaughter pigs are the main reservoir and carcasses are contaminated during a sub-optimal hygienically slaughtering-process. Serology is potentially an easy option to test for the Yersinia-status of the pig (batches) before slaughter. A study of the variation in activity values (OD%) of Yersinia spp. in pigs and pig batches when applying a serological test were therefore conducted. In this study, pieces of the diaphragm of 7047 pigs, originating from 100 farms, were collected and meat juice was gathered, where after an enzyme-linked immunosorbent assay (ELISA) Pigtype Yopscreen (Labor Diagnostik Leipzig, Qiagen, Leipzig, Germany) was performed. The results were defined positive if the activity values exceeded the proposed cut-off value of 30 OD%. Results at pig level displayed a bimodal-shaped distribution with modes at 0-10% (n=879) and 50-60% (n=667). The average OD% was 51% and 66% of the animals tested positive. The within-batch seroprevalence ranged from 0 to 100% and also showed a bimodal distribution with modes at 0% (n=7) and 85-90% (n=16). On 7 farms, no single seropositive animal was present and in 22 farms, the mean OD% was below 30%. Based on the results obtained at slaughter, 66% of the pigs had contact with enteropathogenic Yersinia spp. at farm level. The latter occurred in at least 93% of the farms indicating that most farms are harboring enteropathogenic Yersinia spp.

  16. High Nitrate Concentrations in Vacuolate, Autotrophic Marine Beggiatoa spp

    PubMed Central

    McHatton, S. C.; Barry, J. P.; Jannasch, H. W.; Nelson, D. C.

    1996-01-01

    Massive accumulations of very large Beggiatoa spp. are found at a Monterey Canyon cold seep and at Guaymas Basin hydrothermal vents. Both environments are characterized by high sediment concentrations of soluble sulfide and low levels of dissolved oxygen in surrounding waters. These filamentous, sulfur-oxidizing bacteria accumulate nitrate intracellularly at concentrations of 130 to 160 mM, 3,000- to 4,000-fold higher than ambient levels. Average filament widths range from 24 to 122 (mu)m, and individual cells of all widths possess a central vacuole. These findings plus recent parallel discoveries for Thioploca spp. (H. Fossing, V. A. Gallardo, B. B. Jorgensen, M. Huttel, L. P. Nielsen, H. Schulz, D. E. Canfield, S. Forster, R. N. Glud, J. K. Gundersen, J. Kuver, N. B. Ramsing, A. Teske, B. Thamdrup, and O. Ulloa, Nature (London) 374:713-715, 1995) suggest that nitrate accumulation may be a universal property of vacuolate, filamentous sulfur bacteria. Ribulose bisphosphate carboxylase-oxygenase and 2-oxoglutarate dehydrogenase activities in the Beggiatoa sp. from Monterey Canyon suggest in situ autotrophic growth of these bacteria. Nitrate reductase activity is much higher in the Monterey Beggiatoa sp. than in narrow, laboratory-grown strains of Beggiatoa spp., and the activity is found primarily in the membrane fraction, suggesting that the vacuolate Beggiatoa sp. can reduce nitrate coupled to electron flow through an electron transport system. Nitrate-concentrating and respiration potentials of these chemolithoautotrophs suggest that the Beggiatoa spp. described here are an important link between the sulfur, nitrogen, and carbon cycles at the Monterey Canyon seeps and the Guaymas Basin hydrothermal vents where they are found. PMID:16535282

  17. Bartonella spp. bacteremia in blood donors from Campinas, Brazil.

    PubMed

    Pitassi, Luiza Helena Urso; de Paiva Diniz, Pedro Paulo Vissotto; Scorpio, Diana Gerardi; Drummond, Marina Rovani; Lania, Bruno Grosselli; Barjas-Castro, Maria Lourdes; Gilioli, Rovilson; Colombo, Silvia; Sowy, Stanley; Breitschwerdt, Edward B; Nicholson, William L; Velho, Paulo Eduardo Neves Ferreira

    2015-01-01

    Bartonella species are blood-borne, re-emerging organisms, capable of causing prolonged infection with diverse disease manifestations, from asymptomatic bacteremia to chronic debilitating disease and death. This pathogen can survive for over a month in stored blood. However, its prevalence among blood donors is unknown, and screening of blood supplies for this pathogen is not routinely performed. We investigated Bartonella spp. prevalence in 500 blood donors from Campinas, Brazil, based on a cross-sectional design. Blood samples were inoculated into an enrichment liquid growth medium and sub-inoculated onto blood agar. Liquid culture samples and Gram-negative isolates were tested using a genus specific ITS PCR with amplicons sequenced for species identification. Bartonella henselae and Bartonella quintana antibodies were assayed by indirect immunofluorescence. B. henselae was isolated from six donors (1.2%). Sixteen donors (3.2%) were Bartonella-PCR positive after culture in liquid or on solid media, with 15 donors infected with B. henselae and one donor infected with Bartonella clarridgeiae. Antibodies against B. henselae or B. quintana were found in 16% and 32% of 500 blood donors, respectively. Serology was not associated with infection, with only three of 16 Bartonella-infected subjects seropositive for B. henselae or B. quintana. Bartonella DNA was present in the bloodstream of approximately one out of 30 donors from a major blood bank in South America. Negative serology does not rule out Bartonella spp. infection in healthy subjects. Using a combination of liquid and solid cultures, PCR, and DNA sequencing, this study documents for the first time that Bartonella spp. bacteremia occurs in asymptomatic blood donors. Our findings support further evaluation of Bartonella spp. transmission which can occur through blood transfusions.

  18. Bartonella spp. Bacteremia in Blood Donors from Campinas, Brazil

    PubMed Central

    Pitassi, Luiza Helena Urso; de Paiva Diniz, Pedro Paulo Vissotto; Scorpio, Diana Gerardi; Drummond, Marina Rovani; Lania, Bruno Grosselli; Barjas-Castro, Maria Lourdes; Gilioli, Rovilson; Colombo, Silvia; Sowy, Stanley; Breitschwerdt, Edward B.; Nicholson, William L.; Velho, Paulo Eduardo Neves Ferreira

    2015-01-01

    Bartonella species are blood-borne, re-emerging organisms, capable of causing prolonged infection with diverse disease manifestations, from asymptomatic bacteremia to chronic debilitating disease and death. This pathogen can survive for over a month in stored blood. However, its prevalence among blood donors is unknown, and screening of blood supplies for this pathogen is not routinely performed. We investigated Bartonella spp. prevalence in 500 blood donors from Campinas, Brazil, based on a cross-sectional design. Blood samples were inoculated into an enrichment liquid growth medium and sub-inoculated onto blood agar. Liquid culture samples and Gram-negative isolates were tested using a genus specific ITS PCR with amplicons sequenced for species identification. Bartonella henselae and Bartonella quintana antibodies were assayed by indirect immunofluorescence. B. henselae was isolated from six donors (1.2%). Sixteen donors (3.2%) were Bartonella-PCR positive after culture in liquid or on solid media, with 15 donors infected with B. henselae and one donor infected with Bartonella clarridgeiae. Antibodies against B. henselae or B. quintana were found in 16% and 32% of 500 blood donors, respectively. Serology was not associated with infection, with only three of 16 Bartonella-infected subjects seropositive for B. henselae or B. quintana. Bartonella DNA was present in the bloodstream of approximately one out of 30 donors from a major blood bank in South America. Negative serology does not rule out Bartonella spp. infection in healthy subjects. Using a combination of liquid and solid cultures, PCR, and DNA sequencing, this study documents for the first time that Bartonella spp. bacteremia occurs in asymptomatic blood donors. Our findings support further evaluation of Bartonella spp. transmission which can occur through blood transfusions. PMID:25590435

  19. PCR-DGGE analysis of intestinal bacteria and effect of Bacillus spp. on intestinal microbial diversity in kuruma shrimp ( Marsupenaeus japonicus)

    NASA Astrophysics Data System (ADS)

    Liu, Huaide; Liu, Mei; Wang, Baojie; Jiang, Keyong; Jiang, Shan); Sun, Shujuan; Wang, Lei

    2010-07-01

    In this study, the intestinal microbiota of kuruma shrimp ( Marsupenaeus japonicus) was examined by molecular analysis of the 16S rDNA to identify the dominant intestinal bacteria and to investigate the effects of Bacillus spp. on intestinal microbial diversity. Samples of the intestines of kuruma shrimp fed normal feed and Bacillus spp. amended feed. PCR and denaturing gradient gel electrophoresis (DGGE) analyses were then performed on DNA extracted directly from the guts. Population fingerprints of the predominant organisms were generated by DGGE analysis of the universal V3 16S rDNA amplicons, and distinct bands in the gels were sequenced. The results suggested that the gut of kuruma shrimp was dominated by Vibrio sp. and uncultured gamma proteobacterium. Overall, the results of this study suggest that PCR-DGGE is a possible method of studying the intestinal microbial diversity of shrimp.

  20. Lactic acid fermentation as a tool to enhance the functional features of Echinacea spp

    PubMed Central

    2013-01-01

    Background Extracts and products (roots and/or aerial parts) from Echinacea ssp. represent a profitable market sector for herbal medicines thanks to different functional features. Alkamides and polyacetylenes, phenols like caffeic acid and its derivatives, polysaccharides and glycoproteins are the main bioactive compounds of Echinacea spp. This study aimed at investigating the capacity of selected lactic acid bacteria to enhance the antimicrobial, antioxidant and immune-modulatory features of E. purpurea with the prospect of its application as functional food, dietary supplement or pharmaceutical preparation. Results Echinacea purpurea suspension (5%, wt/vol) in distilled water, containing 0.4% (wt/vol) yeast extract, was fermented with Lactobacillus plantarum POM1, 1MR20 or C2, previously selected from plant materials. Chemically acidified suspension, without bacterial inoculum, was used as the control to investigate functional features. Echinacea suspension fermented with Lb. plantarum C2 exhibited a marked antimicrobial activity towards Gram-positive and -negative bacteria. Compared to control, the water-soluble extract from Echinacea suspension fermented with Lactobacillus plantarum 1MR20 showed twice time higher radical scavenging activity on DPPH. Almost the same was found for the inhibition of oleic acid peroxidation. The methanol extract from Echinacea suspension had inherent antioxidant features but the activity of extract from the sample fermented with strain 1MR20 was the highest. The antioxidant activities were confirmed on Balb 3T3 mouse fibroblasts. Lactobacillus plantarum C2 and 1MR20 were used in association to ferment Echinacea suspension, and the water-soluble extract was subjected to ultra-filtration and purification through RP-FPLC. The antioxidant activity was distributed in a large number of fractions and proportional to the peptide concentration. The antimicrobial activity was detected only in one fraction, further subjected to nano

  1. The Genotypic Characterization of Cronobacter spp. Isolated in China

    PubMed Central

    Cui, Jinghua; Du, Xiaoli; Liu, Hui; Hu, Guangchun; Lv, Guoping; Xu, Baohong; Yang, Xiaorong; Li, Wei; Cui, Zhigang

    2014-01-01

    Cronobacter spp. (Enterobacter sakazakii) is an important pathogen contaminating powdered infant formula (PIF). To describe the genotypic diversity of Cronobacter isolated in China, we identified the isolates using fusA allele sequencing, and subtyped all of the isolates using pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST), and multiple-locus variable-number tandem-repeat analysis (MLVA). A total of 105 isolates were identified, which included C. sakazakii (58 isolates), C. malonaticus (30 isolates), C. dublinensis (11 isolates), C. turicensis (5 isolates), and C. muytjensii (1 isolate). These isolates were showed to have 85 PFGE-patterns, 71 sequence types (STs), and 55 MLVA-patterns. Comparisons among the three molecular subtyping methods revealed that the PFGE method was the most distinguishable tool in identifying clusters of Cronobacter spp. through DNA fingerprinting, and MLST method came second. However, ESTR-1, ESTR-2, ESTR-3, and ESTR-4 were not effective loci for subtyping Cronobacter spp. such that the MLVA method requires further improvement. PMID:25029018

  2. Observations in 2001 on hookworms ( Uncinaria spp.) in otariid pinnipeds.

    PubMed

    Lyons, E T; DeLong, R L; Spraker, T R; Melin, S R; Tolliver, S C

    2003-04-01

    Uncinaria spp. were recovered from the milk of California sea lions ( Zalophus californianus) collected from the: (1) teats of a cow just after parturition (one parasitic third-stage larva, L(3)), (2) stomach of her nursing pup (two L(3)), and (3) stomach of a dead pup about 2 days old (one L(3), one headless, probably L(3), and four L(4)) on San Miguel Island, California in May 2001. This, in addition to earlier research, indicates transmammary transmission of hookworms in this host. Uncinaria spp. were found in dead northern fur seals ( Callorhinus ursinus) in the: (1) intestines of 2 of 75 pups (either one or two adult specimens in each infected pup) and (2) ventral abdominal blubber of 3 of 78 subadult males (one to seven L(3) in each infected seal) on St. Paul Island (SPI), Alaska in July and August 2001. These findings verify the low current prevalence of Uncinariaspp. in fur seals on SPI. Rectal fecal samples taken from 50 live Steller sea lion ( Eumetopias jubatus) pups, about 1 month old, on Rogue Reef in Curry County, Oregon in July 2001, were all negative for the eggs of Uncinaria spp. The apparent zero infection rate in these pups is possibly because the rocky terrain of this rookery is not suitable for hookworm transmission.

  3. Occurrence of Giardia and Cryptosporidium spp. in surface water supplies.

    PubMed Central

    LeChevallier, M W; Norton, W D; Lee, R G

    1991-01-01

    Giardia and Cryptosporidium levels were determined by using a combined immunofluorescence test for source waters of 66 surface water treatment plants in 14 states and 1 Canadian province. The results showed that cysts and oocysts were widely dispersed in the aquatic environment. Giardia spp. were detected in 81% of the raw water samples. Cryptosporidium spp. were found in 87% of the raw water locations. Overall, Giardia or Cryptosporidium spp. were detected in 97% of the raw water samples. Higher cyst and oocyst densities were associated with source waters receiving industrial or sewage effluents. Significant correlations were found between Giardia and Cryptosporidium densities and raw water quality parameters such as turbidity and total and fecal coliform levels. Statistical modeling suggests that cyst and oocyst densities could be predicted on the basis of watershed and water quality characteristics. The occurrence of high levels of Giardia cysts in raw water samples may require water utilities to apply treatment beyond that outlined in the Surface Water Treatment Rule of the U.S. Environmental Protection Agency. PMID:1822675

  4. Vegetable Exudates as Food for Callithrix spp. (Callitrichidae): Exploratory Patterns

    PubMed Central

    Francisco, Talitha Mayumi; Couto, Dayvid Rodrigues; Zanuncio, José Cola; Serrão, José Eduardo; Silva, Ita de Oliveira; Boere, Vanner

    2014-01-01

    Marmosets of the genus Callithrix are specialized in the consumption of tree exudates to obtain essential nutritional resource by boring holes into bark with teeth. However, marmoset preferences for particular tree species, location, type, and other suitable factors that aid in exudate acquisition need further research. In the current study, the intensity of exudate use from Anadenanthera peregrina var. peregrina trees by hybrid marmosets Callithrix spp. groups was studied in five forest fragments in Viçosa, in the state of Minas, Brazil. Thirty-nine A. peregrina var. peregrina trees were examined and 8,765 active and non-active holes were analyzed. The trunk of A. peregrina var. peregrina had a lower number of holes than the canopy: 11% were found on the trunk and 89% were found on the canopy. The upper canopy was the preferred area by Callithrix spp. for obtaining exudates. The intensity of tree exploitation by marmosets showed a moderate-to-weak correlation with diameter at breast height (DBH) and total tree height. The overall results indicate that Anadenanthera peregrina var. peregrina provides food resources for hybrid marmosets (Callithrix spp.) and these animals prefer to explore this resource on the apical parts of the plant, where the thickness, location, and age of the branches are the main features involved in the acquisition of exudates. PMID:25372137

  5. Distribution and characterization of Campylobacter spp. from Russian poultry.

    PubMed

    Stern, N J; Bannov, V A; Svetoch, E A; Mitsevich, E V; Mitsevich, I P; Volozhantsev, N V; Gusev, V V; Perelygin, V V

    2004-02-01

    The distribution of Campylobacter spp. on 13 poultry farms (broiler chicken, quail, pheasant, peacock, and turkey) from eight regions (Vladimir, Vologda, Voronezh, Kaluga, Liptsk, Moscow, Orenburg, and Orel) in Russia was surveyed. Intestinal materials were plated onto Campylobacter-selective medium and plates were incubated microaerobically at 42 degrees C for 24 or 48 h. Identification was based on colonial morphology, microscopic examination, and biochemical tests; latex agglutination assays were used for confirmation. In total, 116 isolates were derived from 370 samples. Isolation rates were similar, regardless of whether the birds were from small or large broiler production farms. Susceptibility of 48 representative (from these production sources) strains of Campylobacter spp. to 38 antimicrobial compounds was determined by disk diffusion assays. All strains tested were sensitive to amikacin, gentamycin, sisomycin, chloramphenicol, imipenem, oleandomycin, erythromycin, azitromycin, and ampicillin. The strains were also sensitive to 100 microg/disk of carbenicillin, fluoroquinolones, and to nitrofurans. Fluoroquinolone sensitivity was most notable and may be related to its limited application in poultry production within Russia. Hippurate and ribosomal RNA gene primers were developed and used to distinguish Campylobacter jejuni and Campylobacter coli and to provide a measure of strain discrimination. The combination of PCR analysis and randomly amplified polymorphic DNA (RAPD) typing were conducted for selected isolates. The various poultry species and the different locations yielded Campylobacter isolates with discrete randomly amplified polymorphic DNA patterns. The distribution and substantial diversity of Campylobacter spp. isolates appears similar to that previously reported in other countries.

  6. Minimum inhibitory concentration of carbapenems and tigecycline against Salmonella spp.

    PubMed

    Capoor, Malini R; Nair, Deepthi; Posti, Jitendra; Singhal, Smita; Deb, Monorama; Aggarwal, Pushpa; Pillai, Parukutty

    2009-03-01

    Antimicrobial resistance in Salmonella spp. is of grave concern, more so in quinolone-resistant and extended-spectrum beta-lactamase (ESBL)-producing isolates that cause complicated infections. The MIC of azithromycin, ciprofloxacin, cefixime, cefepime, ceftriaxone, gatifloxacin, imipenem, levofloxacin, meropenem and ofloxacin (E-test strip) and tigecycline and faropenem (agar dilution) against 210 Salmonella spp. was determined. MIC(90) (defined as the antimicrobial concentration that inhibited growth of 90 % of the strains) of the carbapenems (imipenem and meropenem) for Salmonella Typhi and Salmonella Paratyphi A was 0.064 microg ml(-1). MIC(90) of faropenem was 0.25 microg ml(-1) for S. Typhi, S. Paratyphi A and Salmonella Typhimurium. The MIC(90) of azithromycin for all Salmonella spp. ranged from 8 to 16 microg ml(-1). Tigecycline showed an MIC(90) of 2 microg ml(-1) for S. Typhi, 1 microg ml(-1) for S. Paratyphi A and 4 microg ml(-1) for S. Typhimurium. We concluded that tigecycline and the carbapenems are likely to have roles in the final stage of treatment of quinolone-resistant and ESBL-producing multidrug-resistant salmonellae. PMID:19208884

  7. Occurrence of Yersinia spp. in raw beef, pork and chicken.

    PubMed

    Fukushima, H; Hoshina, K; Nakamura, R; Ito, Y

    1987-04-01

    One hundred and twenty raw ground beef, pork and chicken samples from ten local grocery stores during a one year period were assayed for the presence of Yersinia spp., using direct and postenrichment KOH treatment. Seven thousand and fifty-five different isolates were recovered from 119 (99%) samples of beef and pork and 118 (98%) samples of chicken, including 27 Yersinia enterocolitica serotype 03 biotype 3B phage type 2 (4 pork samples), 4 Y. enterocolitica serotype 03 biotype 4 phage type 8 (3 pork samples), 4,066 Y. enterocolitica biotype 1, 2,194 Yersinia intermedia, 509 Yersinia frederiksenii, and 251 Yersinia kristensenii. Y. enterocolitica serotype 03 was isolated from 6 pork samples, including simultaneously isolation of both biotypes 3B and 4 from one sample. There was the negative correlation between the incidence of isolation of serotype 03 and the environmental temperature, but other environmental Yersinia spp. were frequently isolated through the year. Y. enterocolitica serotype 03 isolates were counted by alkali direct treatment at less than 10(3) cells per g of pork samples, and environmental Yersinia spp. at less than 10(5) cells per g of beef, pork and chicken samples. This may be the first evidence that some pork is contaminated with at least 10(3) Y. enterocolitica serotype 03 cells per g.

  8. Detection of Listeria spp. using ACTERO listeria enrichment media.

    PubMed

    Claveau, David; Olishevskyy, Sergiy; Giuffre, Michael; Martinez, Gabriela

    2014-01-01

    ACTERO Listeria Enrichment Media (ACTERO Listeria) is a selective medium developed for a single-step recovery and enrichment of Listeria spp. from environmental samples. Robustness testing of the ACTERO Listeria medium demonstrated good performance when minor changes were introduced to the incubation temperature and time. All 54 Listeria strains tested, representing the most frequently isolated Listeria species from food (L. monocytogenes, L. ivanovii, L. seeligeri, L. welshimeri, and L. grayi), were successfully enriched in ACTERO Listeria. None of the 30 nontarget strains tested in the exclusivity study was recovered after incubation in ACTERO Listeria. Recovery of Listeria was consistent across three independently produced lots of the ACTERO Listeria, and the prepared medium was stable for 45 days when stored at 4 degrees C in the dark. Matrix studies performed with environmental sponge samples from plastic and stainless steel surfaces demonstrated similar recovery of Listeria spp. in a single-step enrichment using ACTERO Listeria from plastic, and significantly better recovery from stainless steel surfaces when compared to the U.S. Department of Agriculture-Food Safety and Inspection Service reference method. The results of this study prove that ACTERO Listeria Enrichment Media can be effectively used in replacement of the two-step enrichment suggested by the reference method without affecting the recovery of Listeria spp. from environmental samples.

  9. Hemoglobin uptake by Paracoccidioides spp. is receptor-mediated.

    PubMed

    Bailão, Elisa Flávia Luiz Cardoso; Parente, Juliana Alves; Pigosso, Laurine Lacerda; de Castro, Kelly Pacheco; Fonseca, Fernanda Lopes; Silva-Bailão, Mirelle Garcia; Báo, Sônia Nair; Bailão, Alexandre Melo; Rodrigues, Marcio L; Hernandez, Orville; McEwen, Juan G; Soares, Célia Maria de Almeida

    2014-05-01

    Iron is essential for the proliferation of fungal pathogens during infection. The availability of iron is limited due to its association with host proteins. Fungal pathogens have evolved different mechanisms to acquire iron from host; however, little is known regarding how Paracoccidioides species incorporate and metabolize this ion. In this work, host iron sources that are used by Paracoccidioides spp. were investigated. Robust fungal growth in the presence of the iron-containing molecules hemin and hemoglobin was observed. Paracoccidioides spp. present hemolytic activity and have the ability to internalize a protoporphyrin ring. Using real-time PCR and nanoUPLC-MSE proteomic approaches, fungal growth in the presence of hemoglobin was shown to result in the positive regulation of transcripts that encode putative hemoglobin receptors, in addition to the induction of proteins that are required for amino acid metabolism and vacuolar protein degradation. In fact, one hemoglobin receptor ortholog, Rbt5, was identified as a surface GPI-anchored protein that recognized hemin, protoporphyrin and hemoglobin in vitro. Antisense RNA technology and Agrobacterium tumefaciens-mediated transformation were used to generate mitotically stable Pbrbt5 mutants. The knockdown strain had a lower survival inside macrophages and in mouse spleen when compared with the parental strain, which suggested that Rbt5 could act as a virulence factor. In summary, our data indicate that Paracoccidioides spp. can use hemoglobin as an iron source most likely through receptor-mediated pathways that might be relevant for pathogenic mechanisms. PMID:24831516

  10. Vegetable exudates as food for Callithrix spp. (Callitrichidae): exploratory patterns.

    PubMed

    Francisco, Talitha Mayumi; Couto, Dayvid Rodrigues; Zanuncio, José Cola; Serrão, José Eduardo; Silva, Ita de Oliveira; Boere, Vanner

    2014-01-01

    Marmosets of the genus Callithrix are specialized in the consumption of tree exudates to obtain essential nutritional resource by boring holes into bark with teeth. However, marmoset preferences for particular tree species, location, type, and other suitable factors that aid in exudate acquisition need further research. In the current study, the intensity of exudate use from Anadenanthera peregrina var. peregrina trees by hybrid marmosets Callithrix spp. groups was studied in five forest fragments in Viçosa, in the state of Minas, Brazil. Thirty-nine A. peregrina var. peregrina trees were examined and 8,765 active and non-active holes were analyzed. The trunk of A. peregrina var. peregrina had a lower number of holes than the canopy: 11% were found on the trunk and 89% were found on the canopy. The upper canopy was the preferred area by Callithrix spp. for obtaining exudates. The intensity of tree exploitation by marmosets showed a moderate-to-weak correlation with diameter at breast height (DBH) and total tree height. The overall results indicate that Anadenanthera peregrina var. peregrina provides food resources for hybrid marmosets (Callithrix spp.) and these animals prefer to explore this resource on the apical parts of the plant, where the thickness, location, and age of the branches are the main features involved in the acquisition of exudates. PMID:25372137

  11. In vitro susceptibility of Bacillus spp. to selected antimicrobial agents.

    PubMed Central

    Weber, D J; Saviteer, S M; Rutala, W A; Thomann, C A

    1988-01-01

    Although often dismissed as contaminants when isolated from blood cultures, Bacillus spp. are increasingly recognized as capable of causing serious systemic infections. As part of a clinical-microbiological study, 89 strains of Bacillus spp. isolated from clinical blood cultures between 1981 and 1985 had their species determined and were tested for antimicrobial agent susceptibility to 18 antibiotics. Species of isolates were determined by the API 50CH and API 20E systems. Bacillus cereus (54 strains) was the most common species isolated, followed by B. megaterium (13 strains), B. polymyxa (5 strains), B. pumilus (4 strains), B. subtilis (4 strains), B. circulans (3 strains), B. amyloliquefaciens (2 strains), B. licheniformis (1 strain), and Bacillus spp. (3 strains). Microdilution MIC susceptibility tests revealed all B. cereus strains to be susceptible to imipenem, vancomycin, chloramphenicol, gentamicin, and ciprofloxacin. Non-B. cereus strains were most susceptible to imipenem, vancomycin, LY146032, and ciprofloxacin. Disk susceptibility testing suggested that B. cereus was rarely susceptible to penicillins, semisynthetic penicillins, or cephalosporins with the exception of mezlocillin. In contrast, many non-B. cereus strains were susceptible to penicillins, semisynthetic penicillins, and cephalosporins, but marked variability was noted among species. PMID:3395100

  12. Molecular Differentiation of Shigella Spp. from Enteroinvasive E. Coli

    PubMed Central

    Løbersli, I.; Wester, A. L.; Kristiansen, Å.; Brandal, L. T.

    2016-01-01

    A real-time polymerase chain reaction (PCR) assay, amplifying the genes encoding lactose permease (lacY) and invasion plasmid antigen H (ipaH), was run on 121 isolates phenotypically classified as Shigella spp., enteroinvasive Escherichia coli (EIEC), or EIEC O nontypable (ONT). The results were compared with data from a generic E. coli multiple-locus variable-number of tandem repeat analysis (MLVA) and a Shigella MLVA. The real-time PCR verified all Shigella spp. (n = 53) as Shigella (lacY negative) and all EIEC O121 (n = 15) and EIEC O124 (n = 2) as EIEC (lacY positive). However, the real-time PCR typed EIEC O164 as either EIEC (n = 2) or Shigella (n = 2) and, thus, was not suited for classifying this group of isolates. Interestingly, the majority (42/47, 89.4%) of the EIEC ONT were classified as Shigella (lacY negative) by the real-time PCR, and in nearly all cases, (92.9%, 39/42) data from both MLVA assays supported these findings. Overall, in 94.7% (114/121) of the isolates, the results from the real-time PCR were substantiated by the results from the MLVA assays. In conclusion, the real-time PCR assay was fast and accurate in differentiating Shigella spp. from EIEC, with the exception of the EIEC O164 group. This molecular assay was particularly pragmatic for the challenging EIEC ONT group. PMID:27766168

  13. Pinto Bean Yield Increased by Chemical Control of Pratylenchus spp.

    PubMed Central

    Robbins, R. T.; Dickerson, O. J.; Kyle, J. H.

    1972-01-01

    Pinto bean yields and Pratylenchus spp. (nematode) population densities are reported for field plots pro-plant treated with nematicides in 1966 and 1968. Vidden-D (1,3-dichloropropene, 1,2-dichloropropane and related chlorinated hydrocarbons), Vortex (20% methyl isothioeyanate plus 80% chlorinated Ca-hydrocarbons), Telone PBC (80% dichloropropenes, 15% chloropicrin, and 5% propargyl bromide), Dasardt (0,0-Diethyl 0-[p-(methylsulfmyl)phenyl] phosphorothioate, and Dowfume MC-2 (98% methyl bromide plus 2% chloropierin) were used in 1966. Vorlex, Dasanit, and D-D (1,3-dichloropropene, 1,2-dichloropropane and related chlorinated hydrocarbons) were each used at two rates in 1968. Fumigated plot yields ranged 32-56% higher than control plots in 1966 and 11-80% higher in 1968. Significant yield increases were obtained for all fumigants except Telone PBC in 1966. In 1968 significant increases were obtained from use of the high rate (374 liters/ha) of Vorlex and low rate (8.4 liters/ha) of Dasanit. There was an inverse relationship between yield and numbers of Pratylenchus spp./g root on four sampling dates in 1968. A correlation coefficient of -.39 (P ≤ 0.05) was obtained for samples taken 36 days after planting and -.52 (P ≤ 0.01) for samples taken 30 days later. There was no significant correlation between yield and numbers of Pratylenchus spp. recovered from the soil. PMID:19319242

  14. Hemoglobin Uptake by Paracoccidioides spp. Is Receptor-Mediated

    PubMed Central

    Bailão, Elisa Flávia Luiz Cardoso; Parente, Juliana Alves; Pigosso, Laurine Lacerda; de Castro, Kelly Pacheco; Fonseca, Fernanda Lopes; Silva-Bailão, Mirelle Garcia; Báo, Sônia Nair; Bailão, Alexandre Melo; Rodrigues, Marcio L.; Hernandez, Orville; McEwen, Juan G.; Soares, Célia Maria de Almeida

    2014-01-01

    Iron is essential for the proliferation of fungal pathogens during infection. The availability of iron is limited due to its association with host proteins. Fungal pathogens have evolved different mechanisms to acquire iron from host; however, little is known regarding how Paracoccidioides species incorporate and metabolize this ion. In this work, host iron sources that are used by Paracoccidioides spp. were investigated. Robust fungal growth in the presence of the iron-containing molecules hemin and hemoglobin was observed. Paracoccidioides spp. present hemolytic activity and have the ability to internalize a protoporphyrin ring. Using real-time PCR and nanoUPLC-MSE proteomic approaches, fungal growth in the presence of hemoglobin was shown to result in the positive regulation of transcripts that encode putative hemoglobin receptors, in addition to the induction of proteins that are required for amino acid metabolism and vacuolar protein degradation. In fact, one hemoglobin receptor ortholog, Rbt5, was identified as a surface GPI-anchored protein that recognized hemin, protoporphyrin and hemoglobin in vitro. Antisense RNA technology and Agrobacterium tumefaciens-mediated transformation were used to generate mitotically stable Pbrbt5 mutants. The knockdown strain had a lower survival inside macrophages and in mouse spleen when compared with the parental strain, which suggested that Rbt5 could act as a virulence factor. In summary, our data indicate that Paracoccidioides spp. can use hemoglobin as an iron source most likely through receptor-mediated pathways that might be relevant for pathogenic mechanisms. PMID:24831516

  15. Widespread Rickettsia spp. Infections in Ticks (Acari: Ixodoidea) in Taiwan.

    PubMed

    Kuo, Chi-Chien; Shu, Pei-Yun; Mu, Jung-Jung; Lee, Pei-Lung; Wu, Yin-Wen; Chung, Chien-Kung; Wang, Hsi-Chieh

    2015-09-01

    Ticks are second to mosquitoes as the most important disease vectors, and recent decades have witnessed the emergence of many novel tick-borne rickettsial diseases, but systematic surveys of ticks and tick-borne rickettsioses are generally lacking in Asia. We collected and identified ticks from small mammal hosts between 2006 and 2010 in different parts of Taiwan. Rickettsia spp. infections in ticks were identified by targeting ompB and gltA genes with nested polymerase chain reaction. In total, 2,732 ticks were collected from 1,356 small mammals. Rhipicephalus haemaphysaloides Supino (51.8% of total ticks), Haemaphysalis bandicota Hoogstraal & Kohls (28.0%), and Ixodes granulatus Supino (20.0%) were the most common tick species, and Rattus losea Swinhoe (44.7% of total ticks) and Bandicota indica Bechstein (39.9%) were the primary hosts. The average Rickettsia infective rate in 329 assayed ticks was 31.9% and eight Rickettsia spp. or closely related species were identified. This study shows that rickettsiae-infected ticks are widespread in Taiwan, with a high diversity of Rickettsia spp. circulating in the ticks. Because notifiable rickettsial diseases in Taiwan only include mite-borne scrub typhus and flea-borne murine typhus, more studies are warranted for a better understanding of the real extent of human risks to rickettsioses in Taiwan.

  16. Fecal Shedding of Campylobacter and Arcobacter spp. in Dairy Cattle

    PubMed Central

    Wesley, I. V.; Wells, S. J.; Harmon, K. M.; Green, A.; Schroeder-Tucker, L.; Glover, M.; Siddique, I.

    2000-01-01

    Campylobacter jejuni, Campylobacter coli, and Arcobacter spp. were detected in feces of healthy dairy cows by highly specific multiplex-PCR assays. For C. jejuni, at this one-time sampling, cows from 80.6% of farm operations (n = 31) and 37.7% of individual dairy cattle fecal samples (n = 2,085) were positive. Farm management factors were correlated with prevalence in herds in which >25% of cows were positive for C. jejuni. Statistical significance was set at a P of 0.20. Using these criteria, application of manure with broadcast spreaders (P = 0.17), feeding of whole cottonseed or hulls (P = 0.17) or alfalfa (P = 0.15), and accessibility of feed to birds (P = 0.17) were identified as possible risk factors for C. jejuni infection. C. coli was detected in at least one animal in 19.4% of operations and 1.8% of individual cows (n = 2,085). At the herd level, use of broadcaster spreaders was not a risk factor for C. coli infection. For Arcobacter, cows from 71% of dairy operations (n = 31) and 14.3% of individual dairy cattle fecal samples (n = 1,682) were positive. At the herd level, for Arcobacter spp., feeding of alfalfa (P = 0.11) and use of individual waterers (P = 0.19) were protective. This is the first description of Arcobacter spp. in clinically healthy dairy cattle and the first attempt to correlate their presence with C. jejuni. PMID:10788372

  17. Concurrent infections with Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis spp. in naturally infected dairy cattle from birth to two years of age

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fecal specimens were collected directly at weekly and then monthly intervals from each of 30 dairy calves from birth to 24 months to determine the prevalence and age distribution of Cryptosporidium spp., Giardia duodenalis assemblages, Enterocytozoon bieneusi genotypes, and Blastocystis spp subtypes...

  18. Evaluation of a triplex real-time PCR system to detect the plant-pathogenic molds Alternaria spp., Fusarium spp. and C. purpurea.

    PubMed

    Grube, Sabrina; Schönling, Jutta; Prange, Alexander

    2015-12-01

    This article describes the development of a triplex real-time PCR system for the simultaneous detection of three major plant-pathogenic mold genera (Alternaria spp., Fusarium spp. and the species Claviceps purpurea). The designed genus-specific primer-probe systems were validated for sensitivity, specificity and amplification in the presence of background DNA.

  19. Swarming motility is modulated by expression of the putative xenosiderophore transporter SppR-SppABCD in Pseudomonas aeruginosa PA14.

    PubMed

    Pletzer, Daniel; Braun, Yvonne; Weingart, Helge

    2016-06-01

    In the present study, we characterised the putative peptide ABC transporter SppABCD, which is co-transcribed with the TonB-dependent receptor SppR in Pseudomonas aeruginosa PA14. However, our data show that this transporter complex is not involved in the uptake of peptides. The fact that the TonB-dependent receptor SppR is regulated by an iron starvation ECF sigma factor suggested that this transporter is probably involved in the uptake of xenosiderophores. Therefore, we screened culture supernatants of 23 siderophore-producing bacteria for their ability to induce the expression of the SppR-regulating ECF sigma factor. However, none of them had an effect on the expression of this ECF sigma factor. Since the spp operon is not expressed under standard laboratory conditions, we overexpressed it from plasmids in PA14, which led to an impairment of its swarming motility on semisolid agar. Since we excluded the possibility that the uptake of a culture medium component was responsible for the observed phenotype, we hypothesize that the Spp transport system is involved in the uptake of a compound from the periplasmic space or a compound secreted by P. aeruginosa. Furthermore, we found that rhamnolipid synthesis was decreased while biofilm and exopolysaccharide synthesis was slightly increased upon overexpression of the spp operon. Moreover, we observed an impact of spp overexpression on regulation of genes involved in siderophore and phenazine biosynthesis. PMID:26995781

  20. Changes of haematological indices of grass carp, Ceteopharyngodon idella exposed to monogenean parasites, Gyrodactylus spp. and Dactylogyrus spp.

    PubMed

    Restiannasab, Abulhasan; Hemmatzadeh, Mohtaram; Khara, Hossein; Saljoghi, Zoheir Shokouh

    2016-09-01

    The present was carried out to investigate the effects of monogenean infection on haematological indices of grass carp, Ceteopharyngodon idella. In this regard, some haematological indices were measured in two adult groups of grass carp including healthy and infected fish. According to our results, the values of red blood cells (RBCs), haemoglobin (Hb) decreased significantly in infected fishes (P < 0.05). In contrast, the white blood cells (WBCs) values increased significantly in infected fishes (P < 0.05). In contrast, the WBC values increased significantly in infected fishes. In conclusion, our results showed that monogenean infection by Gyrodactylus spp. and Dactylogyrus spp. can affects health condition of grass carp through alternation of haematology. PMID:27605756

  1. Prevalence of Salmonella spp. and thermophilic Campylobacter spp. in the small Asian mongoose (Herpestes javanicus) in Barbados, West Indies.

    PubMed

    Rhynd, Kamara J R; Leighton, Patrick A; Elcock, David A; Whitehall, Pamela J; Rycroft, Andrew; Macgregor, Shaheed K

    2014-12-01

    From April to July 2005, rectal swabs were collected from 48 free-ranging small Asian mongooses (Herpestes javanicus) on the east and south coasts of Barbados and analyzed for Salmonella and Campylobacter spp. Salmonella was recovered in 21.12% (7/33) of mongooses at the east-coast site and 26.67% (4/15) at the south-coast site. Four serotypes were isolated: Salmonella enterica serovar Rubislaw, Kentucky, Javiana, and Panama. One east-coast sample of 11 tested for Campylobacter was positive (9.09%). These results indicate that mongooses in Barbados are carriers and shedders of Salmonella and Campylobacter spp. and are a potential wildlife reservoir for these enteropathogens. PMID:25632681

  2. Analysis of phenolic compounds and radical scavenging activity of Echinacea spp.

    PubMed

    Pellati, Federica; Benvenuti, Stefania; Magro, Lara; Melegari, Michele; Soragni, Fabrizia

    2004-04-16

    The aim of this study was to set up and validate an RP-LC method with DAD-detection to quantify caffeic acid derivatives in various Echinacea spp. Samples were extracted with 80% methanol. The analyses were carried out on a Lichrospher RP-18 column (125 mm x 4 mm i.d., 5 microm), with a mobile phase gradient, which increases the acetonitrile level in a phosphoric acid solution (0.1%). The flow rate was 1.5 ml/min. Detection was set at 330 nm. This method allowed the identification and quantification of caftaric acid, chlorogenic acid, caffeic acid, cynarin, echinacoside and cichoric acid in Echinacea roots and derivatives. The total phenolic content was 10.49 mg/g for E. angustifolia, 17.83 mg/g for E. pallida and 23.23 mg/g for E. purpurea. Among Echinacea commercial herbal medicines, a certain variability in the concentrations of phenolic compounds was observed. The radical scavenging activity of Echinacea methanolic extracts was evaluated in vitro with a spectrophotometric method based on the reduction of an alcoholic 2,2-diphenyl-1-picrylhydrazyl (DPPH*) radical solution at 517 nm in the presence of a hydrogen donating antioxidant. As for pure compounds, echinacoside had the highest capacity to quench DPPH* radicals (EC50 = 6.6 microM), while caftaric acid had the lowest (EC50 = 20.5 microM). The average EC50 values for E. purpurea, E. pallida and E. angustifolia were 134, 167 and 231 microg/ml, respectively. The radical scavenging activity of Echinacea root extracts reflected their phenolic composition. The results indicate that Echinacea roots and derivatives are a good source of natural antioxidants and could be used to prevent free-radical-induced deleterious effects.

  3. Alicyclobacillus spp. in the fruit juice industry: history, characteristics, and current isolation/detection procedures.

    PubMed

    Chang, Su-Sen; Kang, Dong-Hyun

    2004-01-01

    The first Alicyclobacillus spp. was isolated in 1982, and was originally thought to be strictly limited to thermophilic and acidic environments. Two years later, another Alicyclobacillus sp., A. acidoterrestris, was identified as the causative agent in spoilage of commercially pasteurized apple juice. Subsequent studies soon found that Alicyclobacillus spp. are soilborne bacteria, and do not strictly require thermophilic and acidic environments. Alicyclobacillus spp. posess several distinct characteristics; the major one is their ability to survive commercial pasteurization processes and produce off-flavors in fruit juices. The fruit juice industry has acknowledged Alicyclobacillus spp. as a major quality control target microorganism. Guaiacol and halophenols were identified as the offensive smelling agent in many Alicyclobacillus spp. related spoilage. Though the exact formation pathway of these off-flavors by Alicyclobacillus spp. are not yet identified, studies report that the presence of Alicyclobacillus spp. in the medium may be a major contributor to the formation of these off-flavors. Many identification methods and isolation media were developed in the last two decades. However, most of these methods were developed specifically for A. acidoterrestris, which was the first identified off-flavor producing Alicyclobacillus. However, recent studies indicate that other species of Alicyclobacillus may also produce guaiacol or the halophenols. In this respect, all Alicyclobacillus spp. should be monitored as potential spoilage bacteria in fruit juices. This article includes an overall review of the history of Alicyclobacillus spp., characteristics, suggested off-flavor production pathways, and commonly used identification methods for the currently identified Alicyclobacillus spp.

  4. First detection of Cytauxzoon spp. infection in European wildcats (Felis silvestris silvestris) of Italy.

    PubMed

    Veronesi, Fabrizia; Ravagnan, Silvia; Cerquetella, Matteo; Carli, Erika; Olivieri, Emanuela; Santoro, Azzurra; Pesaro, Stefano; Berardi, Sara; Rossi, Giacomo; Ragni, Bernardino; Beraldo, Paola; Capelli, Gioia

    2016-07-01

    Cytauxzoonosis is an emerging, tick-transmitted, protozoan disease affecting domestic and wild felids and caused by Cytauxzoon felis, Cytauxzoon manul and Cytauxzoon spp. This study aimed to determine the presence of infection with Cytauxzoon spp. in Felis silvestris silvestris in Italy, in order to enhance the comprehension of its pattern distribution among domestic cat populations. In addition, wildcats were tested for other endemic vector-borne pathogens in Italy. The carcasses of 21 F. s. silvestris were collected from central and northern regions of Italy. All the animals were submitted to necropsy and samples of the spleens were collected. Cytauxzoon infection was surveyed by a conventional PCR amplifying a portion of the SSU-rDNA of species of Piroplasmida. The samples were also screened for Anaplasma spp., Ehrlichia spp., Rickettsia spp., Babesia spp., Theileria spp., and Leishmania spp. using SYBR Green Real-Time PCR (rPCR) assays. Four animals (19%) were positive for Piroplasmida-PCR assay and three sequenced amplicons were obtained (14.3%), clustering with the Italian, Spanish, French and Romanian Cytauxzoon spp. isolates and with C. manul found in Mongolia. The samples were negative for the other pathogens screened. The present results showed that Cytauxzoon spp. may infect both F. s. silvestris and F. s. catus. PMID:27150590

  5. Morphological, Chemical, and Genetic Diversity of Tropical Marine Cyanobacteria Lyngbya spp. and Symploca spp. (Oscillatoriales)†

    PubMed Central

    Thacker, Robert W.; Paul, Valerie J.

    2004-01-01

    Although diverse natural products have been isolated from the benthic, filamentous cyanobacterium Lyngbya majuscula, it is unclear whether this chemical variation can be used to establish taxonomic relationships among disparate collections. We compared morphological characteristics, secondary-metabolite compositions, and partial 16S ribosomal DNA (rDNA) sequences among several collections of L. majuscula Gomont, Lyngbya spp., and Symploca spp. from Guam and the Republic of Palau. The morphological characteristics examined were cell length, cell width, and the presence or absence of a calyptra. Secondary metabolites were analyzed by two-dimensional thin-layer chromatography. Each collection possessed a distinct cellular morphology that readily distinguished Lyngbya spp. from Symploca spp. Each collection yielded a unique chemotype, but common chemical characteristics were shared among four collections of L. majuscula. A phylogeny based on secondary-metabolite composition supported the reciprocal monophyly of Lyngbya and Symploca but yielded a basal polytomy for Lyngbya. Pairwise sequence divergence among species ranged from 10 to 14% across 605 bp of 16S rDNA, while collections of L. majuscula showed 0 to 1.3% divergence. Although the phylogeny of 16S rDNA sequences strongly supported the reciprocal monophyly of Lyngbya and Symploca as well as the monophyly of Lyngbya bouillonii and L. majuscula, genetic divergence was not correlated with chemical and morphological differences. These data suggest that 16S rDNA sequence analyses do not predict chemical variability among Lyngbya species. Other mechanisms, including higher rates of evolution for biosynthetic genes, horizontal gene transfer, and interactions between different genotypes and environmental conditions, may play important roles in generating qualitative and quantitative chemical variation within and among Lyngbya species. PMID:15184125

  6. Fast and discriminative CoSYPS detection system of viable Salmonella spp. and Listeria spp. in carcass swab samples.

    PubMed

    Barbau-Piednoir, Elodie; Botteldoorn, Nadine; Mahillon, Jacques; Dierick, Katelijne; Roosens, Nancy H

    2015-01-01

    In this study, the complete CoSYPS Path Food workflow including all steps, namely swab sample enrichment, SYBR®Green qPCR detection of Salmonella spp. and Listeria spp., isolation and confirmation of the detected strain, was validated on beef carcass swabs. To perform the validation, the results of the complete workflow were compared, according to the ISO 16140:2003, with the ISO reference methods for detection, isolation and confirmation of Listeria monocytogenes and Salmonella spp. The results showed that the relative level of detection and the limit of detection of the complete workflow and ISO reference methods are in a range from 2 to 16 CFU/swab for both bacteria. The relative specificity, sensitivity and accuracy identified during this validation were all 100% since the results obtained with the complete CoSYPS Path Food workflow and the ISO reference methods were identical (Cohen's kappa index=1.00). In addition the complete CoSYPS Path Food workflow is able to provide detection results (negative or presumptive positive) in half the time needed as for the ISO reference methods. These results demonstrate that the performance of the complete CoSYPS Path Food workflow is not only comparable to the ISO reference methods but also provides a faster response for the verification of beef carcasses before commercial distribution.

  7. Phenolic profiling of the South American "Baylahuen" tea (Haplopappus spp., Asteraceae) by HPLC-DAD-ESI-MS.

    PubMed

    Schmeda-Hirschmann, Guillermo; Quispe, Cristina; González, Benita

    2015-01-01

    The aerial parts of several Haplopappus species (Asteraceae), known under the common name "baylahuen", are used as herbal teas in Chile and Argentina. In Chile, "baylahuen" comprises H. multifolius, H. taeda, H. baylahuen and H. rigidus. Little is known about the chemical identity of the infusion constituents in spite of widespread consumption. The aim of the present work was the characterization of phenolics occurring in the infusions and methanol extracts of "baylahuen" by HPLC-DAD-ESI-MS. A simple HPLC-DAD-ESI-MS method was developed for the fast identification and differentiation of Haplopappus spp. used as a tea source, based on the phenolics from the tea and methanol extracts. Some 27 phenolics were tentatively identified in the infusions and methanol extract, including 10 caffeoyl quinic and feruloyl quinic acid derivatives and 17 flavonoids. The HPLC patterns of the Haplopappus tea and methanol extract allow a clear differentiation at the species level. The occurrence of hydroxycinnamic acid derivatives and flavonoids can explain the reputed nutraceutical and health beneficial properties of this herbal tea. PMID:25580687

  8. Universal extraction method for gastrointestinal pathogens.

    PubMed

    Halstead, Fenella D; Lee, Adele V; Couto-Parada, Xose; Polley, Spencer D; Ling, Clare; Jenkins, Claire; Chalmers, Rachel M; Elwin, Kristin; Gray, Jim J; Iturriza-Gómara, Miren; Wain, John; Clark, Duncan A; Bolton, Frederick J; Manuel, Rohini J

    2013-10-01

    A universal stool extraction method for recovery of nucleic acids (NAs) from gastrointestinal pathogens was developed to support rapid diagnostics for the London 2012 Olympics. The method involved mechanical disruption (bead beating) of the stools, followed by automated extraction and detection using real-time PCR. This method had been used extensively in the Second Infectious Intestinal Disease Study (IID2) for the isolation of NA from bacteria and parasites (and was effective for the robust recovery of Cryptosporidium spp.) but had not been used for enteric viruses. To ensure this method was universally suitable, panels of samples known to contain target bacteria, viruses or parasites were processed in triplicate using the pre-treatment method routinely used for each target and the new extraction method (bead beating). The extracts were tested using real-time PCR and the cycle threshold values were compared. The results from this study showed that bead beating improved yields for the bacterial and parasitic targets and was suitable for the viral targets. The implementation of this universal method should confer cost- and time-saving benefits and streamline the processes required for the characterization of an array of pathogens from faecal samples. PMID:23831766

  9. An ELISA for detecting anti-Plasmodium spp. antibodies in African black-footed penguins (Spheniscus demersus).

    PubMed

    Graczyk, T K; Cranfield, M R; Skjoldager, M L; Shaw, M L

    1994-02-01

    An enzyme-linked immunosorbent assay (ELISA) utilizing 3 Plasmodium falciparum antigens, R32tet32, P.F.R27, and crude red blood cell extract, was developed for the detection of circulating anti-Plasmodium relictum or anti-Plasmodium elongatum antibodies in sera from naturally infected adult African black-footed penguins (Spheniscus demersus) at The Baltimore Zoo, Maryland. A concentration of 2.0 micrograms/ml of each antigen was optimal in terms of specificity, sensitivity, and test speed. It was possible to detect anti-Plasmodium spp. antibodies at a dilution of 10(-4.11). Low absorbance values (less than 0.050) of nonspecific background were observed. The binding efficacy of anti-penguin IgG coupled to alkaline phosphatase to antibodies in the penguin sera was significantly higher than the binding efficacy of anti-chicken IgG. All penguins, bled in the winter time, in controlled mosquito-free conditions had anti-Plasmodium spp. antibodies reactive with P. falciparum antigens. The penguins showed age-dependent variation in antibody levels. There was a decrease in antibody titration units that was significantly correlated with the number of outdoor exposure years experienced by the birds, despite the season-comparable epizootiologic conditions in their summer open-air habitat. We concluded that the decrease of anti-malarial antibodies could be explained by an antibody-mediated equilibrium of immunity in naturally immunized birds harboring endothelial-stage parasites. The ELISA described is sensitive, and it requires a minimal amount of equipment to collect the blood samples. The assay can be used for detecting and monitoring levels of anti-Plasmodium spp. antibodies in selected groups of penguins.

  10. LUMINEX®: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools

    PubMed Central

    2013-01-01

    Background Six species of the genus Entamoeba, i.e., E. histolytica, E. dispar, E. moshkovskii, E. polecki, E. coli, and E. hartmanii can be found in human stools. Among these, only E. histolytica is considered to be pathogenic, causing intestinal and extra-intestinal disease, but it is morphologically identical to E. dispar and E. moshkovskii. In general, E. polecki, E. coli, and E. hartmanii can be differentiated morphologically from E. histolytica, but some of their diagnostic morphologic features may overlap creating issues for the differential diagnosis. Moreover, the previous inability to differentiate among Entamoeba species has limited epidemiologic information on E histolytica. The objective of this study was to develop a rapid, high-throughput screening method using Luminex technique for the simultaneous detection and differentiation of Entamoeba species. Methods PCR amplification was performed with biotinylated Entamoeba sp 18S rRNA gene primers, designed to amplify a fragment ranging from 382 to 429 bp of the Entamoeba spp studied. Regions of this fragment that could differentiate among E. histolytica, E. moshkovskii, E. dispar, E. hartmanii and E. coli were selected to design hybridization probes to link to Luminex beads. The assay was standardized with cloned DNA samples of each species and evaluated with 24 DNA extracts from samples obtained from individuals diagnosed with these amebas in their stools. Results Using this approach we were able to correctly identify E. histoltyica, E. dispar, E hartmanni, E. coli and E. moshkovskii in all specimens studied. From twenty four samples tested by microscopy, PCR/DNA Sequencing and real-time PCR, 100% agreed with PCR-Luminex assay for identification of E. dispar, E. moshkovskii, E. hartmanni, E. histolytica, and E. coli. Conclusion These results show that this method could be used in the diagnostic detection of Entamoeba spp in fecal samples. This diagnostic test was useful to clearly distinguish E

  11. Ruminal Prevotella spp. May Play an Important Role in the Conversion of Plant Lignans into Human Health Beneficial Antioxidants

    PubMed Central

    Schogor, Ana L. B.; Huws, Sharon A.; Santos, Geraldo T. D.; Scollan, Nigel D.; Hauck, Barbara D.; Winters, Ana L.; Kim, Eun J.; Petit, Hélène V.

    2014-01-01

    Secoisolariciresinol diglucoside (SDG), the most abundant lignan in flaxseed, is metabolized by the ruminal microbiota into enterolignans, which are strong antioxidants. Enterolactone (EL), the main mammalian enterolignan produced in the rumen, is transferred into physiological fluids, with potentially human health benefits with respect to menopausal symptoms, hormone-dependent cancers, cardiovascular diseases, osteoporosis and diabetes. However, no information exists to our knowledge on bacterial taxa that play a role in converting plant lignans into EL in ruminants. In order to investigate this, eight rumen cannulated cows were used in a double 4×4 Latin square design and fed with four treatments: control with no flax meal (FM), or 5%, 10% and 15% FM (on a dry matter basis). Concentration of EL in the rumen increased linearly with increasing FM inclusion. Total rumen bacterial 16S rRNA concentration obtained using Q-PCR did not differ among treatments. PCR-T-RFLP based dendrograms revealed no global clustering based on diet indicating between animal variation. PCR-DGGE showed a clustering by diet effect within four cows that had similar basal ruminal microbiota. DNA extracted from bands present following feeding 15% FM and absent with no FM supplementation were sequenced and it showed that many genera, in particular Prevotella spp., contributed to the metabolism of lignans. A subsequent in vitro study using selected pure cultures of ruminal bacteria incubated with SDG indicated that 11 ruminal bacteria were able to convert SDG into secoisolariciresinol (SECO), with Prevotella spp. being the main converters. These data suggest that Prevotella spp. is one genus playing an important role in the conversion of plant lignans to human health beneficial antioxidants in the rumen. PMID:24709940

  12. Apparatus for hydrocarbon extraction

    SciTech Connect

    Bohnert, George W.; Verhulst, Galen G.

    2013-03-19

    Systems and methods for hydrocarbon extraction from hydrocarbon-containing material. Such systems and methods relate to extracting hydrocarbon from hydrocarbon-containing material employing a non-aqueous extractant. Additionally, such systems and methods relate to recovering and reusing non-aqueous extractant employed for extracting hydrocarbon from hydrocarbon-containing material.

  13. [Research advances in the effects of environmental factors on the growth and development of Aurelia spp].

    PubMed

    Wang, Jian-Yan; Yu, Zhi-Gang; Zhen, Yu; Mi, Tie-Zhu; Yao, Qing-Zhen; Wang, Guo-Shan

    2012-11-01

    Aurelia spp. is a cosmopolitan coastal species, and also, one dominant species of large jellyfish in the coastal waters of China. In recent years, Aurelia spp. bloom events occur frequently in the world, causing severe damage to marine ecosystems, coastal economy, and society development. Aurelia spp. has a complicated life history comprising a benthic asexually-reproducing polyp generation and a sexually-reproducing medusa generation, and various vegetative reproduction (budding, strobilation, and podocyst production) and sexual reproduction. Surrounding physical and biological factors affect each growth stage of Aurelia spp., especially the juvenile stage of planktonic-benthic life cycle, which has major effect on the population dynamics of Aurelia spp. This paper reviewed the research advances in the effects of environmental factors on Aurelia spp. at its different growth and development stages, and discussed some problems worthy of further study, aimed to provide useful reference for the research of the key factors controlling the jellyfish blooms in coastal waters of China.

  14. Development of a test system for rapid differentiation of Neisseria and Haemophilus spp.

    PubMed Central

    Eriquez, L A; Hodinka, N E

    1983-01-01

    A qualitative micromethod (IDS Rapid NH system) employing conventional and single-substrate enzyme tests was developed for the biochemical characterization of Neisseria spp., Haemophilus spp., and other gram-negative species. A total of over 140 dehydrated, miniaturized biochemical tests were investigated for their ability to distinguish species. Computer-assisted test selection and pair separation analysis of the data allowed the selection of 11 4-h tests that would identify Haemophilus and Neisseria spp. implicated as etiological agents as well as differentiate them from other Neisseria spp., Moraxella spp., Branhamella catarrhalis, Centers for Disease Control M groups, and Kingella spp. The final test configuration included modified glucose, sucrose, galactosidase, nitrate, phosphatase, resazurin reduction, and two arylamidase tests. In addition, indole, urea, and ornithine decarboxylase tests were included to biochemically type strains of Haemophilus influenzae and Haemophilus parainfluenzae. PMID:6358247

  15. Desmodus rotundus and Artibeus spp. bats might present distinct rabies virus lineages.

    PubMed

    Fahl, Willian Oliveira; Carnieli, Pedro; Castilho, Juliana Galera; Carrieri, Maria Luiza; Kotait, Ivanete; Iamamoto, Keila; Oliveira, Rafael Novaes; Brandão, Paulo Eduardo

    2012-01-01

    In Brazil, bats have been assigned an increasing importance in public health as they are important rabies reservoirs. Phylogenetic studies have shown that rabies virus (RABV) strains from frugivorous bats Artibeus spp. are closely associated to those from the vampire bat Desmodus rotundus, but little is known about the molecular diversity of RABV in Artibeus spp. The N and G genes of RABV isolated from Artibeus spp. and cattle infected by D. rotundus were sequenced, and phylogenetic trees were constructed. The N gene nucleotides tree showed three clusters: one for D. rotundus and two for Artibeus spp. Regarding putative N amino acid-trees, two clusters were formed, one for D. rotundus and another for Artibeus spp. RABV G gene phylogeny supported the distinction between D. rotundus and Artibeus spp. strains. These results show the intricate host relationship of RABV's evolutionary history, and are invaluable for the determination of RABV infection sources.

  16. Desmodus rotundus and Artibeus spp. bats might present distinct rabies virus lineages.

    PubMed

    Fahl, Willian Oliveira; Carnieli, Pedro; Castilho, Juliana Galera; Carrieri, Maria Luiza; Kotait, Ivanete; Iamamoto, Keila; Oliveira, Rafael Novaes; Brandão, Paulo Eduardo

    2012-01-01

    In Brazil, bats have been assigned an increasing importance in public health as they are important rabies reservoirs. Phylogenetic studies have shown that rabies virus (RABV) strains from frugivorous bats Artibeus spp. are closely associated to those from the vampire bat Desmodus rotundus, but little is known about the molecular diversity of RABV in Artibeus spp. The N and G genes of RABV isolated from Artibeus spp. and cattle infected by D. rotundus were sequenced, and phylogenetic trees were constructed. The N gene nucleotides tree showed three clusters: one for D. rotundus and two for Artibeus spp. Regarding putative N amino acid-trees, two clusters were formed, one for D. rotundus and another for Artibeus spp. RABV G gene phylogeny supported the distinction between D. rotundus and Artibeus spp. strains. These results show the intricate host relationship of RABV's evolutionary history, and are invaluable for the determination of RABV infection sources. PMID:23146155

  17. Chemical Composition and Biological Activities of Fragrant Mexican Copal (Bursera spp.).

    PubMed

    Gigliarelli, Giulia; Becerra, Judith X; Curini, Massimo; Marcotullio, Maria Carla

    2015-01-01

    Copal is the Spanish word used to describe aromatic resins from several genera of plants. Mexican copal derives from several Bursera spp., Protium copal, some Pinus spp. (e.g., P. pseudostrobus) and a few Fabaceae spp. It has been used for centuries as incense for religious ceremonies, as a food preservative, and as a treatment for several illnesses. The aim of this review is to analyze the chemical composition and biological activity of commercial Mexican Bursera copal. PMID:26703535

  18. Chemical Composition and Biological Activities of Fragrant Mexican Copal (Bursera spp.).

    PubMed

    Gigliarelli, Giulia; Becerra, Judith X; Curini, Massimo; Marcotullio, Maria Carla

    2015-12-12

    Copal is the Spanish word used to describe aromatic resins from several genera of plants. Mexican copal derives from several Bursera spp., Protium copal, some Pinus spp. (e.g., P. pseudostrobus) and a few Fabaceae spp. It has been used for centuries as incense for religious ceremonies, as a food preservative, and as a treatment for several illnesses. The aim of this review is to analyze the chemical composition and biological activity of commercial Mexican Bursera copal.

  19. Molecular Characterization of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in Captive Wildlife at Zhengzhou Zoo, China.

    PubMed

    Li, Junqiang; Qi, Meng; Chang, Yankai; Wang, Rongjun; Li, Tongyi; Dong, Haiju; Zhang, Longxian

    2015-01-01

    Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are common gastrointestinal protists in humans and animals. Two hundred and three fecal specimens from 80 wildlife species were collected in Zhengzhou Zoo and their genomic DNA extracted. Three intestinal pathogens were characterized with a DNA sequence analysis of different loci. Cryptosporidium felis, C. baileyi, and avian genotype III were identified in three specimens (1.5%), the manul, red-crowned crane, and cockatiel, respectively. Giardia duodenalis was also found in five specimens (2.5%) firstly: assemblage B in a white-cheeked gibbon and beaver, and assemblage F in a Chinese leopard and two Siberian tigers, respectively. Thirteen genotypes of E. bieneusi (seven previously reported genotypes and six new genotypes) were detected in 32 specimens (15.8%), of which most were reported for the first time. A phylogenetic analysis of E. bieneusi showed that five genotypes (three known and two new) clustered in group 1; three known genotypes clustered in group 2; one known genotype clustered in group 4; and the remaining four genotypes clustered in a new group. In conclusion, zoonotic Cryptosporidium spp., G. duodenalis, and E. bieneusi are maintained in wildlife and transmitted between them. Zoonotic disease outbreaks of these infectious agents possibly originate in wildlife reservoirs.

  20. Genetic diversity and expression of the [NiFe] hydrogenase large-subunit gene of Desulfovibrio spp. in environmental samples.

    PubMed Central

    Wawer, C; Jetten, M S; Muyzer, G

    1997-01-01

    The genetic diversity and expression of the [NiFe] hydrogenase large-subunit gene of Desulfovibrio spp. in environmental samples were determined in order to show in parallel the existing and active members of Desulfovibrio populations. DNA and total RNA were extracted from different anaerobic bioreactor samples; RNA was transcribed into cDNA. Subsequently, PCR was performed to amplify a ca.-440-bp fragment of the [NiFe] hydrogenase large-subunit gene and its mRNA. Denaturing gradient gel electrophoresis analysis was used to separate the PCR products according to their sequence and thereby to visualize the individual community members. Desulfovibrio strains corresponding to amplified [NiFe] hydrogenase transcripts were regarded as metabolically active, because in pure cultures transcripts were detectable in exponentially growing cells but not in cultures in the stationary phase. DNA sequencing and comparative sequence analysis were used to identify the detected organisms on the basis of their [NiFe] hydrogenase sequences. The genes of characterized Desulfovibrio spp. showed a considerable extent of divergence (ca. 30%), whereas sequences obtained from bacterial populations of the bioreactors showed a low level of variation and indicated the coexistence of closely related strains probably belonging to the species Desulfovibrio sulfodismutans. Under methanogenic conditions, all detected populations were active; under denitrifying conditions, no [NiFe] hydrogenase mRNA was visible. Changes in activity and composition of Desulfovibrio populations caused by changes in the environmental conditions could be monitored by using the approach described in this study. PMID:9361423

  1. Sequestration of Dimethylsulfoniopropionate (DMSP) and Acrylate from the Green Alga Ulva Spp. by the Sea Hare Aplysia juliana.

    PubMed

    Kamio, Michiya; Koyama, Mao; Hayashihara, Nobuko; Hiei, Kaori; Uchida, Hajime; Watanabe, Ryuichi; Suzuki, Toshiyuki; Nagai, Hiroshi

    2016-05-01

    Many animals sequester secondary metabolites from their food. In this study, we hypothesized that the sea hare Aplysia juliana sequesters secondary metabolites from green algae. To test this, we performed NMR-based metabolomic analysis on methanol extracts of Ulva spp. and A. juliana. Another sea hare, Bursatella leachii, which mainly feeds on another type of alga, was added to this analysis as an outgroup. Two body parts of the sea hares, skin and digestive glands, were used in the analysis. Principal component analysis (PCA) on the NMR data of these samples detected biomarkers common to Ulva spp. and A. juliana. This result indicates sequestration of secondary metabolites by the herbivore from the plants. The biomarker metabolites were identified as dimethylsulfoniopropionate (DMSP) and acrylate, which were concentrated in skin of A. juliana and were released from the skin of live animals when physically stressed. Thus, our NMR-based metabolomic study revealed sequestration of algae-derived secondary metabolites in skin of A. Juliana, and in the discharge of the metabolites under conditions that mimic attack by predators. PMID:27179528

  2. Infections of Hypostomus spp. by Trypanosoma spp. and leeches: a study of hematology and record of these hirudineans as potential vectors of these hemoflagellates.

    PubMed

    Corrêa, Lincoln Lima; Oliveira, Marcos Sidney Brito; Tavares-Dias, Marcos; Ceccarelli, Paulo Sérgio

    2016-01-01

    Among Kinetoplastida, the Trypanosoma is the genus with the highest occurrence infecting populations of marine fish and freshwater in the world, with high levels of prevalence, causing influences fish health and consequent economic losses, mainly for fish populations in situation stress. This study investigated infections of Hypostomus spp. by Trypanosoma spp. and leeches, as well as blood parameters of this host in the network of tributaries of the Tapajós River in the state of Pará, in the eastern Amazon region in Brazil. Of the 47 hosts examined, 89.4% were parasitized by Trypanosoma spp. and 55.4% also had leeches attached around the mouth. The intensity of Trypanosoma spp. increased with the size of the host, but the body conditions were not influenced by the parasitism. The number of red blood cells, and hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH), total number of leukocytes and thrombocytes showed variations and negative correlation with the intensity of Trypanosoma spp. in the blood of the hosts. The results suggest that the leeches were vectors of Trypanosoma spp. in Hypostomus spp. PMID:27580397

  3. Duplex PCR for detection of Salmonella and Shigella spp in cockle samples.

    PubMed

    Senachai, Pachara; Chomvarin, Chariya; Wongboot, Warawan; Boonyanugomol, Wongwarut; Tangkanakul, Waraluk

    2013-09-01

    Salmonella and Shigella spp are important causative agents of foodborne diseases. A sensitive, specific and rapid method is essential for detection of these pathogens. In this study, a duplex PCR method was developed for simultaneous detection of Salmonella and Shigella spp in cockle samples and compared with the traditional culture method. Enrichment broths for Salmonella spp recovery were also compared. Sensitivity of the duplex PCR for simultaneous detection of Salmonella and Shigella spp from pure culture was 10(3) CFU/ml (40 CFU/PCR reaction), and that of sterile cockle samples spiked with these two pathogens was 1 CFU/10 g of cockle tissue after 9 hours enrichment [3 hours in buffered peptone water (BPW), followed by 6 hours in Rappaport Vasiliadis (RV) broth or tetrathionate (TT) broth for Salmonella spp and 6 hours enrichment in Shigella broth (SB) for Shigella spp]. There was no significant difference in detection sensitivity between enrichment in RV and TT broths. Salmonella spp detected in cockles in Khon Kaen, Thailand by duplex PCR and culture method was 17% and 13%, respectively but Shigella spp was not detected. The duplex PCR technique developed for simultaneous detection of Salmonella and Shigella spp in cockle samples was highly sensitive, specific and rapid and could serve as a suitable method for food safety assessment.

  4. Molecular tracking of Salmonella spp. in chicken meat chain: from slaughterhouse reception to end cuts.

    PubMed

    Dias, Mariane Rezende; Cavicchioli, Valéria Quintana; Camargo, Anderson Carlos; Lanna, Frederico Germano Piscitelli Alvarenga; Pinto, Paulo Sérgio de Arruda; Bersot, Luciano Dos Santos; Nero, Luís Augusto

    2016-02-01

    Due to the importance of Salmonella spp. in poultry products, this study aimed to track its main contamination routes since slaughtering reception to processing of chicken end cuts. Samples from different steps of slaughtering and processing (n = 277) were collected from two chicken slaughterhouses (Sl1 and Sl2) located in Minas Gerais state, Brazil, and subjected to Salmonella spp. detection. The obtained isolates were subjected to serological identification and tested by PCR for specific Salmonella spp. genes (ompC and sifB). Also, Salmonella spp. isolates were subjected to XbaI macrorestriction and pulsed-field gel electrophoresis (PFGE). Sixty-eight samples were positive for Salmonella spp. and 172 isolates were obtained. Sl1 and Sl2 presented similar frequencies of Salmonella spp. positive samples during reception, slaughtering and processing (p > 0.05), except for higher frequencies in Sl1 for chicken carcasses after de-feathering and evisceration (p < 0.05). PFGE allowed the identification of cross contamination and persistence of Salmonella spp. strains in Sl1. The results highlighted the relevance of the initial steps of chicken slaughtering for Salmonella spp. contamination, and the pre-chilling of carcasses as an important controlling tool. In addition, the presence of Salmonella spp. in chicken end cuts samples represents a public health concern. PMID:27162388

  5. Differential Legionella spp. survival between intracellular and extracellular forms in thermal spring environments.

    PubMed

    Kao, Po-Min; Tung, Min-Che; Hsu, Bing-Mu; Hsu, Shih-Yung; Huang, Jen-Te; Liu, Jorn-Hon; Huang, Yu-Li

    2013-05-01

    Legionella are commonly found in natural and man-made aquatic environments and are able to inhabit various species of protozoa. The relationship between the occurrence of Legionella spp. within protozoa and human legionellosis has been demonstrated; however, the proportions of intracellular and extracellular Legionella spp. in the aquatic environment were rarely reported. In this study, we developed a new method to differentiate intracellular and extracellular Legionella spp. in the aquatic environment. Water samples from three thermal spring recreational areas in southeastern Taiwan were collected and analyzed. For each water sample, concurrent measurements were performed for Legionella spp. and their free-living amoebae hosts. The overall detection rate was 32 % (16/50) for intracellular Legionella spp. and 12 % (6/50) for extracellular Legionella spp. The most prevalent host of Legionella spp. was Hartmannella vermiformis. The identified Legionella spp. differed substantially between intracellular and extracellular forms. The results showed that it may be necessary to differentiate intracellular and extracellular forms of Legionella spp.

  6. Presence of Contracaecum spp. in teleosts cultured and fished in Sardinia.

    PubMed

    Salati, Fulvio; Meloni, Mauro; Cau, Manuela; Angelucci, Giulia

    2013-09-23

    This study reports the results of the finding of Contracaecum spp. during a survey on endoparasites isolated from cultured and wild fish and also from some cephalopods caught in Sardinian waters. Contracaecum spp. is a nematode belonging to the Anisakidae, and is reported to cause zoonosis in humans. Nematodes were detected after visual inspection and enzymatic digestion and then identified by morphologic observation, which was confirmed by PCR. The results show that Contracaecum spp. were found in both fish caught from sea or lagoon, and in both cultured and wild fish: 33 of the parasitized samples were wild fish (24 caught in the sea and 9 in lagoons) and 11 were cultured ones. The prevalence of Contracaecum spp. was higher in Diplodus spp. (16.0%), Sparus aurata (15.8%) and Mullus spp. (14.6%). Larvae were also found by enzymatic digestion at muscular level in 5 species, with the highest prevalence in S. aurata (10.5%). The results of this study indicate that Contracaecum spp. was present in cultured fish such as S. aurata, Diplodus spp. and Dicentrarchus labrax. All cultured fish with parasites were collected from land-based semi-intensive tanks whose water came from an adjacent lagoon. Finally, the evidence that this parasite is found in both cultured and wild fish leads us to re-consider the zoonotic potential of Contracaecum spp., in particular when one bears in mind its dimensions at the L3 stage, when it is barely visible to the human eye.

  7. Duplex PCR for detection of Salmonella and Shigella spp in cockle samples.

    PubMed

    Senachai, Pachara; Chomvarin, Chariya; Wongboot, Warawan; Boonyanugomol, Wongwarut; Tangkanakul, Waraluk

    2013-09-01

    Salmonella and Shigella spp are important causative agents of foodborne diseases. A sensitive, specific and rapid method is essential for detection of these pathogens. In this study, a duplex PCR method was developed for simultaneous detection of Salmonella and Shigella spp in cockle samples and compared with the traditional culture method. Enrichment broths for Salmonella spp recovery were also compared. Sensitivity of the duplex PCR for simultaneous detection of Salmonella and Shigella spp from pure culture was 10(3) CFU/ml (40 CFU/PCR reaction), and that of sterile cockle samples spiked with these two pathogens was 1 CFU/10 g of cockle tissue after 9 hours enrichment [3 hours in buffered peptone water (BPW), followed by 6 hours in Rappaport Vasiliadis (RV) broth or tetrathionate (TT) broth for Salmonella spp and 6 hours enrichment in Shigella broth (SB) for Shigella spp]. There was no significant difference in detection sensitivity between enrichment in RV and TT broths. Salmonella spp detected in cockles in Khon Kaen, Thailand by duplex PCR and culture method was 17% and 13%, respectively but Shigella spp was not detected. The duplex PCR technique developed for simultaneous detection of Salmonella and Shigella spp in cockle samples was highly sensitive, specific and rapid and could serve as a suitable method for food safety assessment. PMID:24437322

  8. Expression and purification of bioactive high-purity recombinant mouse SPP1 in Escherichia coli.

    PubMed

    Yuan, Yunsheng; Zhang, Xiyuan; Weng, Shunyan; Guan, Wen; Xiang, Di; Gao, Jin; Li, Jingjing; Han, Wei; Yu, Yan

    2014-05-01

    Secreted phosphoprotein 1 (SPP1) is a phosphorylated acidic glycoprotein. It is broadly expressed in a variety of tissues, and it is involved in a number of physiological and pathological events, including cancer metastasis, tissues remodeling, pro-inflammation regulation, and cell survival. SPP1 has shown its function of protecting tissues and organs against injury and wound, giving itself potentials to become a therapy target or giving its antibodies of other counter-acting reagents potentials to become drug candidates. Non-tagged (native) recombinant SPP1 would be valuable in therapeutic and pharmaceutical researches. In our study, mouse Spp1 DNA fragment without signal peptide was built in pET28a(+) vector and transformed into Escherichia coli BL21 (DE3). The recombinant mouse SPP1 (rmSPP1) was then expressed in bacteria upon induction by isopropyl β-D-thiogalactopyranoside (IPTG). The abundance of rmSPP1 was increased using isoelectric precipitation and ammonium sulfate fractionation methods, and anion and cation exchange chromatography was employed to further purify rmSPP1. Finally, we got rmSPP1 product with 12.8 % productivity, 97 % purity, satisfactory bioactivity, and low endotoxin content. PMID:24664233

  9. Comparative Study of Hydroalcoholic Extracts of Momordica charantia L. against Foodborne Pathogens.

    PubMed

    Rakholiya, Kalpna; Vaghela, P; Rathod, T; Chanda, Sumitra

    2014-03-01

    The antimicrobial effect of 24 different hydroalcoholic extracts (100, 75, 50 and 25% methanol and water) obtained from four parts (leaf+stem (aerial), peel, pulp and seed) of Momordica charantia L. were investigated against five Gram-positive, six Gram-negative and four fungal strains. The extraction was done by individual cold percolation method using hexane, different hydroalcoholic solvent (100, 75, 50 and 25% methanol) and water. The antimicrobial activity was done by agar well diffusion assay. The extracts, which showed >15 mm zone of inhibition, were further screened to determine minimum inhibitory concentration and minimum bactericidal concentration using a broth dilution method performed in 96-well microtitre plate. The extractive yield was highest in aqueous extracts of all the four parts closely followed by 25% methanol. Micrococcus flavus was the most susceptible Gram-positive bacteria and Pseudomonas testosteroni was the most susceptible Gram-negative bacteria. The highest antibacterial activity was shown by 100% methanol. The Gram-negative Pseudomonas spp. was more susceptible towards all the extracts than the Gram-positive bacteria or fungal strains investigated. One hundred percent and 50% methanol extracts of seed showed lowest minimum inhibitory concentration and minimum bactericidal concentration values, that is <39 and 625 μg/ml, respectively, against Pseudomonas pictorum. Therefore, these extracts would be of interest in the control of Pseudomonas spp. in food industry as well as used for therapeutic purposes. PMID:24843188

  10. Comparative Study of Hydroalcoholic Extracts of Momordica charantia L. against Foodborne Pathogens

    PubMed Central

    Rakholiya, Kalpna; Vaghela, P.; Rathod, T.; Chanda, Sumitra

    2014-01-01

    The antimicrobial effect of 24 different hydroalcoholic extracts (100, 75, 50 and 25% methanol and water) obtained from four parts (leaf+stem (aerial), peel, pulp and seed) of Momordica charantia L. were investigated against five Gram-positive, six Gram-negative and four fungal strains. The extraction was done by individual cold percolation method using hexane, different hydroalcoholic solvent (100, 75, 50 and 25% methanol) and water. The antimicrobial activity was done by agar well diffusion assay. The extracts, which showed >15 mm zone of inhibition, were further screened to determine minimum inhibitory concentration and minimum bactericidal concentration using a broth dilution method performed in 96-well microtitre plate. The extractive yield was highest in aqueous extracts of all the four parts closely followed by 25% methanol. Micrococcus flavus was the most susceptible Gram-positive bacteria and Pseudomonas testosteroni was the most susceptible Gram-negative bacteria. The highest antibacterial activity was shown by 100% methanol. The Gram-negative Pseudomonas spp. was more susceptible towards all the extracts than the Gram-positive bacteria or fungal strains investigated. One hundred percent and 50% methanol extracts of seed showed lowest minimum inhibitory concentration and minimum bactericidal concentration values, that is <39 and 625 μg/ml, respectively, against Pseudomonas pictorum. Therefore, these extracts would be of interest in the control of Pseudomonas spp. in food industry as well as used for therapeutic purposes. PMID:24843188

  11. Transcriptional profile of Paracoccidioides spp. in response to itraconazole

    PubMed Central

    2014-01-01

    Background Itraconazole is currently used to treat paracoccidioidomycosis. The mechanism of action of azoles has been elucidated in some fungi, although little is known regarding its mechanism of action in Paracoccidioides spp. The present work focused on identification of regulated transcripts using representational difference analysis of Paracoccidioides spp. yeast cells treated with itraconazole for 1 and 2 h. Results Paracoccidioides Pb01 genes up-regulated by itraconazole included genes involved in cellular transport, metabolism/energy, transcription, cell rescue, defense and virulence. ERG11, ERG6, ERG3, ERG5 and ERG25 were up-regulated at multiple time points. In vivo infection experiments in mice corroborated the in vitro results. Ergosterol levels and distribution were evaluated in Paracoccidioides Pb18 yeast cells, and the results demonstrate that both factors were changed in the fungus treated with itraconazole. Conclusion To our knowledge, this is the first transcriptional analysis of Paracoccidioides spp. exposed to a triazole drug. Here acetyl seems to be intensively produced from different metabolic pathways to produce ergosterol by the action of ergosterol synthesis related enzymes, which were also affected in other fungi. Among the genes affected, we identified genes in common with other fungi, as well as genes unique to Paracoccidioides Pb01. Those genes could be considered target to new drugs. Voltage-gated Ca2+ alpha subunit (CAV), Tetracycline resistance protein (TETA) and Hemolisyn-iii channel protein (HLYiii) were found only here and a probably involvement with resistence to itraconazole could be investigated in the future. However our findings do not permit inference to current clinical practice. PMID:24690401

  12. [Comparison of different methods in order to identify Proteus spp].

    PubMed

    Castro, S T; Rodríguez, C R; Perazzi, B E; Radice, M; Paz Sticott, M; Muzio, H; Juárez, J; Gutkind, G; Famiglietti, A M R; Santini, P I; Vay, C A

    2006-01-01

    Comparison of different methods in order to identify Proteus spp. The objectives were: (a) to identify Proteus strains to species level, following Farmer's and O'Hara's conventional biochemical reactions; b) to evaluate the sensitivity and specificity of both the API 20E method and a schema of reduced reactions (TSI and MIO agar: motility, indole and ornithine) comparing them with conventional methodology, and c) to evaluate the utility of SDS-PAGE (total proteins) in order to identify Proteus strains to species level. Two hundred and five Proteus spp. clinical isolates, were collected between January 1998 and September 2004, from inpatients and outpatients at Hospital de Clinicas. Strains were identified by means of conventional methodology, the API 20E method, and a schema of reduced reactions. SDS-PAGE (total proteins) was used in 48 out of the 205 strains. The API 20E method identified 79 out of 87 (90.8%) strains of P. mirabilis, 103 out of 103 P. vulgaris complex, and 15 out of 15 P. penneri. Eight strains of P. mirabilis were identified as Proteus spp., the acid production from maltose being necessary to identify them to species level. The schema of reduced reactions identified 205 out of 205 (100%) strains, that is, this schema of reduced reactions identified all the strains to species level without any additional tests, in marked contrast to the API 20E method. The SDS-PAGE (total proteins) identified the three species of the genus, even if the strains of P. mirabilis showed different biochemical reactions. PMID:17152651

  13. Diversity and antibiotic resistance in Pseudomonas spp. from drinking water.

    PubMed

    Vaz-Moreira, Ivone; Nunes, Olga C; Manaia, Célia M

    2012-06-01

    Pseudomonas spp. are common inhabitants of aquatic environments, including drinking water. Multi-antibiotic resistance in clinical isolates of P. aeruginosa is widely reported and deeply characterized. However, the information regarding other species and environmental isolates of this genus is scant. This study was designed based on the hypothesis that members of the genus Pseudomonas given their high prevalence, wide distribution in waters and genetic plasticity can be important reservoirs of antibiotic resistance in drinking water. With this aim, the diversity and antibiotic resistance phenotypes of Pseudomonas isolated from different drinking water sources were evaluated. The genotypic diversity analyses were based on six housekeeping genes (16S rRNA, rpoD, rpoB, gyrB, recA and ITS) and on pulsed field gel electrophoresis. Susceptibility to 21 antibiotics of eight classes was tested using the ATB PSE EU (08) and disk diffusion methods. Pseudomonas spp. were isolated from 14 of the 32 sampled sites. A total of 55 non-repetitive isolates were affiliated to twenty species. Although the same species were isolated from different sampling sites, identical genotypes were never observed in distinct types of water (water treatment plant/distribution system, tap water, cup fillers, biofilm, and mineral water). In general, the prevalence of antibiotic resistance was low and often the resistance patterns were related with the species and/or the strain genotype. Resistance to ticarcillin, ticarcillin with clavulanic acid, fosfomycin and cotrimoxazol were the most prevalent (69-84%). No resistance to piperacillin, levofloxacin, ciprofloxacin, tetracycline, gentamicin, tobramycin, amikacin, imipenem or meropenem was observed. This study demonstrates that Pseudomonas spp. are not so widespread in drinking water as commonly assumed. Nevertheless, it suggests that water Pseudomonas can spread acquired antibiotic resistance, preferentially via vertical transmission.

  14. Frequency and Spatial Distribution of Environmental Campylobacter spp.

    PubMed Central

    Brown, P. E.; Christensen, O. F.; Clough, H. E.; Diggle, P. J.; Hart, C. A.; Hazel, S.; Kemp, R.; Leatherbarrow, A. J. H.; Moore, A.; Sutherst, J.; Turner, J.; Williams, N. J.; Wright, E. J.; French, N. P.

    2004-01-01

    Humans are exposed to Campylobacter spp. in a range of sources via both food and environmental pathways. For this study, we explored the frequency and distribution of thermophilic Campylobacter spp. in a 10- by 10-km square rural area of Cheshire, United Kingdom. The area contains approximately 70, mainly dairy, farms and is used extensively for outdoor recreational activities. Campylobacter spp. were isolated from a range of environmental samples by use of a systematic sampling grid. Livestock (mainly cattle) and wildlife feces and environmental water and soil samples were cultured, and isolates were presumptively identified by standard techniques. These isolates were further characterized by PCR. Campylobacter jejuni was the most prevalent species in all animal samples, ranging from 11% in samples from nonavian wildlife to 36% in cattle feces, and was isolated from 15% of water samples. Campylobacter coli was commonly found in water (17%) and sheep (21%) samples, but rarely in other samples. Campylobacter lari was recovered from all sample types, with the exception of sheep feces, and was found in moderate numbers in birds (7%) and water (5%). Campylobacter hyointestinalis was only recovered from cattle (7%) and birds (1%). The spatial distribution and determinants of C. jejuni in cattle feces were examined by the use of model-based spatial statistics. The distribution was consistent with very localized within-farm or within-field transmission and showed little evidence of any larger-scale spatial dependence. We concluded that there is a potentially high risk of human exposure to Campylobacter spp., particularly C. jejuni, in the environment of our study area. The prevalence and likely risk posed by C. jejuni-positive cattle feces in the environment diminished as the fecal material aged. After we took into account the age of the fecal material, the absence or presence of rain, and the presence of bird feces, there was evidence of significant variation in the

  15. Trichinella spp. imported with live animals and meat.

    PubMed

    Pozio, Edoardo

    2015-09-30

    Nematodes of the genus Trichinella are widely distributed throughout the world in omnivorous and carnivorous animals (mammals, birds, and reptiles) and in incidental hosts. To prevent the transmission of these zoonotic parasites to humans, meat samples from Trichinella spp. susceptible animals are tested at the slaughterhouse or in game processing plants. The aim of the present review was to collect documented cases on Trichinella infected animals, meat, or meat derived products which reached the international trade or were illegally introduced from one to another country in personal baggage. In the course of the last 60 years in the international literature, there have been 43 reports of importation of Trichinella spp. infected animals or meat, most of which (60%, 26/43) related to live horses or their meat. Meat or meat derived products from pigs, wild boar and bears, account only for 18.6% (8/43), 4.7% (3/43), and 14.3% (6/43), respectively. However, only live horses or their meat intended for human consumption, meat from a single wild boar, and live polar bears caught in the wild for zoos, were imported through the international market; whereas, meat from pigs, wild boars and bears were illegally introduced in a country in personal baggage. Trichinella infected animals or meat which were officially or illegally introduced in a country were the source of 3443 Trichinella infections in humans in a 40-year period (1975-2014). Most of these infections (96.8%) have been linked to horsemeat consumption, whereas meat from pigs, wild boars and bears accounted only for 2.2%, 0.7% and 0.3% of cases, respectively. This review shows the Trichinella spp. risk in the international animal and meat trade has been linked mainly to horses and only one time to wild boar, if they carcasses are not adequately tested, whereas pigs and other wild animals or their derived products infected with Trichinella spp. are unlikely to reach the international market by the official animal and

  16. Atoxoplasma spp. infection in captive canaries (Serinus canaria).

    PubMed

    Sánchez-Cordón, P J; Gómez-Villamandos, J C; Gutiérrez, J; Sierra, M A; Pedrera, M; Bautista, M J

    2007-02-01

    Clinical signs, histopathological and ultrastructural findings associated with Atoxoplasma spp. natural infection in captive canaries (Serinus canaria) are described. Intracytoplasmic Atoxoplasma-like protozoa were found in the liver and lung. In the liver, protozoa were found in hepatocytes and Kupffer's cells and were associated with granulomatous hepatitis and a marked bile duct hyperplasia. An usual finding was the presence of infected mononuclear cells adhered to the endothelium of the blood vessels in lung. The diagnosis was confirmed by ultrastructural examination of reprocessed paraffin-embedded tissues.

  17. Selective medium for isolation and enumeration of Bifidobacterium spp.

    PubMed Central

    Muñoa, F J; Pares, R

    1988-01-01

    A new method was developed for the isolation and enumeration of Bifidobacterium spp. from natural aquatic environments. The method was based on the utilization of a new medium, Bifidobacterium iodoacetate medium 25, and resuscitation techniques were used to isolate injured bifidobacteria. The new medium was tested with a nonselective reference medium on sewage and sewage-polluted surface waters. Relatively little colonial growth of any other bacterial genera occurred; when such colonies did grow, Bifidobacterium could be easily differentiated by its colonial morphology or, after Gram staining, by its typical bifidobacterial morphology. PMID:3415235

  18. Mitochondrial transcripts and associated heteroplasmies of Ancistrus spp. (Siluriformes: Loricariidae)

    PubMed Central

    Moreira, Daniel A.; Furtado, Carolina; Parente, Thiago E.

    2015-01-01

    This data-set complements our paper entitled “The use of transcriptomic next-generation sequencing data to assembly mitochondrial genomes of Ancistrus spp. (Loricariidae)” [6]. Here, we present the nucleotide sequences of each transcript used for mitogenomes assembly, as well as tables presenting the location of each transcript in the mitogenomes; the frequency, location and codon position of the detected heteroplasmic sites; and the start/stop codons usage, UTR, CDS and poliA-tail length for each protein coding gene. Readers are referred to the paper cited above for data interpretation and discussion. PMID:26629496

  19. Mitochondrial transcripts and associated heteroplasmies of Ancistrus spp. (Siluriformes: Loricariidae).

    PubMed

    Moreira, Daniel A; Furtado, Carolina; Parente, Thiago E

    2015-12-01

    This data-set complements our paper entitled "The use of transcriptomic next-generation sequencing data to assembly mitochondrial genomes of Ancistrus spp. (Loricariidae)" [6]. Here, we present the nucleotide sequences of each transcript used for mitogenomes assembly, as well as tables presenting the location of each transcript in the mitogenomes; the frequency, location and codon position of the detected heteroplasmic sites; and the start/stop codons usage, UTR, CDS and poliA-tail length for each protein coding gene. Readers are referred to the paper cited above for data interpretation and discussion. PMID:26629496

  20. Serologic survey for Trichinella spp. in grizzly bears from Alaska.

    PubMed

    Zarnke, R L; Gamble, R; Heckert, R A; Ver Hoef, J

    1997-07-01

    Blood was collected from 878 grizzly bears (Ursus arctos) in seven geographic areas of Alaska from 1973 to 1987. An enzyme-linked immunosorbent assay procedure was used to test sera for evidence of exposure to Trichinella spp. Serum antibody prevalence ranged from 5% (10 positive of 196 tested) in the Southern Region of the state to 83% (355 of 430 tested) in the Northern Region. These major discrepancies may be a result of differing food habits of bears in the major geographic areas. Prevalence was higher in older age cohorts. Neither year-of-collection nor sex had a significant effect on prevalence.

  1. Trichinella spp. imported with live animals and meat.

    PubMed

    Pozio, Edoardo

    2015-09-30

    Nematodes of the genus Trichinella are widely distributed throughout the world in omnivorous and carnivorous animals (mammals, birds, and reptiles) and in incidental hosts. To prevent the transmission of these zoonotic parasites to humans, meat samples from Trichinella spp. susceptible animals are tested at the slaughterhouse or in game processing plants. The aim of the present review was to collect documented cases on Trichinella infected animals, meat, or meat derived products which reached the international trade or were illegally introduced from one to another country in personal baggage. In the course of the last 60 years in the international literature, there have been 43 reports of importation of Trichinella spp. infected animals or meat, most of which (60%, 26/43) related to live horses or their meat. Meat or meat derived products from pigs, wild boar and bears, account only for 18.6% (8/43), 4.7% (3/43), and 14.3% (6/43), respectively. However, only live horses or their meat intended for human consumption, meat from a single wild boar, and live polar bears caught in the wild for zoos, were imported through the international market; whereas, meat from pigs, wild boars and bears were illegally introduced in a country in personal baggage. Trichinella infected animals or meat which were officially or illegally introduced in a country were the source of 3443 Trichinella infections in humans in a 40-year period (1975-2014). Most of these infections (96.8%) have been linked to horsemeat consumption, whereas meat from pigs, wild boars and bears accounted only for 2.2%, 0.7% and 0.3% of cases, respectively. This review shows the Trichinella spp. risk in the international animal and meat trade has been linked mainly to horses and only one time to wild boar, if they carcasses are not adequately tested, whereas pigs and other wild animals or their derived products infected with Trichinella spp. are unlikely to reach the international market by the official animal and

  2. Keratitis due to Histoplasma spp. in a horse.

    PubMed

    Richter, Marianne; Hauser, Beat; Kaps, Simone; Spiess, Bernhard M

    2003-06-01

    A 5-year-old Holsteiner gelding from Germany was presented 2 months after a whitish discoloration of the left cornea was observed. Cytologic examination revealed intra- and extracellular globular structures, up to 4 micro m in size, consisting of a central spherical deeply basophilic body surrounded by an unstained halo. The structures were morphologically consistent with Histoplasma spp. Infection with Histoplasma organisms is not endemic in Europe. Topical use of fluconazole was successful in eliminating Histoplasma organisms within 10 days of initiation of treatment. PMID:12753609

  3. Standardization of Weed Pollen Extracts, Japanese Hop and Mugwort, in Korea

    PubMed Central

    Jeong, Kyoung Yong; Son, Mina; Choi, Soo-Young; Park, Kyung Hee; Park, Hye Jung; Hong, Chein-Soo; Lee, Jae-Hyun

    2016-01-01

    Purpose Japanese hop (Humulus spp.) and mugwort (Artemisia spp.) are notable causes of autumn pollinosis in East Asia. However, Japanese hop and mugwort pollen extracts, which are widely used for the diagnosis, have not been standardized. This study was performed to standardize Japanese hop and mugwort pollen extracts. Materials and Methods Allergen extracts were prepared in a standardized way using locally collected Humulus japonicus and purchased Artemisia vulgaris pollens. The immunoglobulin E (IgE) reactivities of prepared extracts were compared with commercial extracts via IgE immunoblotting and inhibition analyses. Intradermal skin tests were performed to determine the bioequivalent allergy unit (BAU). Results The IgE reactive components of the extracts via IgE immunoblotting were similar to those of commercial extracts. A 11-kDa allergen showed the strongest IgE reactivity in Japanese hop, as did a 28-kDa allergen in mugwort pollen extracts. Allergenic potencies of the investigatory Japanese hop and mugwort extracts were essentially indistinguishable from the commercial ones. Sums of erythema of 50 mm by the intradermal skin test (ΣED50) were calculated to be 14.4th and 13.6th three-fold dilutions for Japanese hop and mugwort extracts, respectively. Therefore, the allergenic activity of the prepared extracts was 90827.4 BAU/mg for Japanese hop and 34412 BAU/mg for mugwort. Conclusion We produced Japanese hop and mugwort pollen extracts using a standardized method. Standardized Japanese hop and mugwort pollen extracts will facilitate the production of improved diagnostic and immunotherapeutic reagents. PMID:26847293

  4. Morphometric and molecular analysis of mackerel (Rastrelliger spp) from the west coast of Peninsular Malaysia.

    PubMed

    Darlina, M N; Masazurah, A R; Jayasankar, P; Jamsari, A F J; Siti, A M N

    2011-01-01

    Mackerel (Scombridae; Rastrelliger) are small commercially important pelagic fish found in tropical regions. They serve as a cheap source of animal protein and are commonly used as live bait. By using a truss morphometrics protocol and RAPD analysis, we examined morphological and genetic variation among 77 individual mackerel that were caught using long lines and gillnets at 11 locations along the west coast of Peninsular Malaysia. Nineteen morphometric traits were evaluated and genetic information was estimated using five 10-base RAPD random primers. Total DNA was extracted from muscle tissue. Morphometric discriminant function analysis revealed that two morphologically distinct groups of Rastrelliger kanagurta and a single group of R. brachysoma can be found along the west coast of Peninsular Malaysia. We also found that the head-related characters and those from the anterior part of the body of Rastrelliger spp significantly contribute to stock assessment of this population. RAPD analysis showed a trend similar to that of the morphometric analysis, suggesting a genetic component to the observed phenotypic differentiation. These data will be useful for developing conservation strategies for these species. PMID:21968625

  5. Pollen-stigma adhesion in Brassica spp involves SLG and SLR1 glycoproteins.

    PubMed Central

    Luu, D T; Marty-Mazars, D; Trick, M; Dumas, C; Heizmann, P

    1999-01-01

    The adhesion of pollen grains to the stigma is the first step of pollination in flowering plants. During this step, stigmas discriminate between pollen grains that can and cannot be permitted to effect fertilization. This selection is operated by various constituents of the cell walls of both partners. Several genes structurally related to the self-incompatibility system that prevents self-pollination in Brassica spp are known to target their products into the stigma cell wall. We proposed previously that one of these genes, the one encoding the S locus glycoprotein (SLG)-like receptor 1 (SLR1), which is coexpressed with that encoding SLG, may participate in pollen-stigma adhesion. Here, we exploit a biomechanical assay to measure the pollen adhesion force and show that it is reduced both by transgenic suppression of SLR1 expression and by pretreatment of wild-type stigmas with anti-SLR1 antibodies, anti-SLG antibodies, or pollen coat-protein extracts. Our results indicate a common adhesive function for the SLR1 and SLG proteins in the pollination process. PMID:9927642

  6. Macrolactams: a novel class of antifungal antibiotics produced by Actinomadura spp. SCC 1776 and SCC 1777.

    PubMed

    Hegde, V; Patel, M; Horan, A; Gullo, V; Marquez, J; Gunnarsson, I; Gentile, F; Loebenberg, D; King, A; Puar, M

    1992-05-01

    Three novel antifungal antibiotics, Sch 38518, Sch 39185 and Sch 38516 were isolated from the fermentation broths of two actinomycetes identified by chemical, morphological and physiological analysis as a new species of Actinomadura. The compounds were isolated from broth by solvent extraction and purified by silica gel chromatography. Physico-chemical properties, mass spectral analysis, IR and UV suggested the compounds were similar. Sch 38518 and Sch 39185 have a molecular formula of C25H48N2O5. 1H NMR, 13C NMR and hydrolysis indicated the aglycones were identical, however the compounds differed in containing isomeric sugar moieties. Sch 38518 contains mycosamine while Sch 39185 contains 3,6-dideoxy-3-amino-L-talopyranose. Sch 38516 has a molecular formula of C24H46N2O5 and is a lower homolog of Sch 39185. The three compounds, Sch 38518 (1), Sch 39185 (2), and Sch 38516 (3) exhibit similar activity against Candida spp. with geometric mean MICs of 1.81, 2.00 and 0.91 micrograms/ml, respectively. PMID:1624364

  7. Bioremediation potential of glyphosate-degrading Pseudomonas spp. strains isolated from contaminated soil.

    PubMed

    Zhao, Haoyu; Tao, Ke; Zhu, Jianyi; Liu, Shengnan; Gao, Han; Zhou, Xiaogang

    2015-01-01

    Bacterial strains capable of utilizing glyphosate as the sole carbon source were isolated from contaminated soil by the enrichment culture method and identified based on partial 16S rRNA gene sequence analysis. Pseudomonas spp. strains GA07, GA09 and GC04 demonstrated the best degradation capabilities towards glyphosate and were used for the laboratory experiments of glyphosate bioremediation. Inoculating glyphosate-treated soil samples with these three strains resulted in a 2-3 times higher rate of glyphosate removal than that in non-inoculated soil. The degradation kinetics was found to follow a first-order model with regression values greater than 0.96. Cell numbers of the introduced bacteria decreased from 4.4 × 10(6) CFU/g to 3.4-6.7 × 10(5) CFU/g dry soil within 18 days of inoculation. Due to the intense degradation of glyphosate, the total dehydrogenase activity of the soil microbial community increased by 21.2-25.6%. Analysis of glyphosate degradation products in cell-free extracts showed that glyphosate breakdown in strain GA09 was catalyzed both by C-P lyase and glyphosate oxidoreductase. Strains GA07 and GC04 degraded glyphosate only via glyphosate oxidoreductase, but no further metabolite was detected. These results highlight the potential of the isolated bacteria to be used in the bioremediation of GP-contaminated soils. PMID:26582285

  8. European validation of Real-Time PCR method for detection of Salmonella spp. in pork meat.

    PubMed

    Delibato, Elisabetta; Rodriguez-Lazaro, David; Gianfranceschi, Monica; De Cesare, Alessandra; Comin, Damiano; Gattuso, Antonietta; Hernandez, Marta; Sonnessa, Michele; Pasquali, Frédérique; Sreter-Lancz, Zuzsanna; Saiz-Abajo, María-José; Pérez-De-Juan, Javier; Butrón, Javier; Prukner-Radovcic, Estella; Horvatek Tomic, Danijela; Johannessen, Gro S; Jakočiūnė, Džiuginta; Olsen, John E; Chemaly, Marianne; Le Gall, Francoise; González-García, Patricia; Lettini, Antonia Anna; Lukac, Maja; Quesne, Segolénè; Zampieron, Claudia; De Santis, Paola; Lovari, Sarah; Bertasi, Barbara; Pavoni, Enrico; Proroga, Yolande T R; Capuano, Federico; Manfreda, Gerardo; De Medici, Dario

    2014-08-01

    The classical microbiological method for detection of Salmonella spp. requires more than five days for final confirmation, and consequently there is a need for an alternative methodology for detection of this pathogen particularly in those food categories with a short shelf-life. This study presents an international (at European level) ISO 16140-based validation study of a non-proprietary Real-Time PCR-based method that can generate final results the day following sample analysis. It is based on an ISO compatible enrichment coupled to an easy and inexpensive DNA extraction and a consolidated Real-Time PCR assay. Thirteen laboratories from seven European Countries participated to this trial, and pork meat was selected as food model. The limit of detection observed was down to 10 CFU per 25 g of sample, showing excellent concordance and accordance values between samples and laboratories (100%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (100%) when the results obtained for the Real-Time PCR-based methods were compared to those of the ISO 6579:2002 standard method. The results of this international trial demonstrate that the evaluated Real-Time PCR-based method represents an excellent alternative to the ISO standard. In fact, it shows an equal and solid performance as well as it reduces dramatically the extent of the analytical process, and can be easily implemented routinely by the Competent Authorities and Food Industry laboratories. PMID:24513055

  9. Cryptococcus spp. isolation from excreta of pigeons (Columba livia) in and around Monterrey, Mexico.

    PubMed

    Canónico-González, Yolanda; Adame-Rodríguez, Juan Manuel; Mercado-Hernández, Roberto; Aréchiga-Carvajal, Elva Teresa

    2013-01-01

    The presence of Cryptococcus spp. has been reported in Mexico's capital city; however, to our knowledge there are no reports of its presence in the state of Nuevo León located in northeast Mexico. This is presumed to be because the hot and dry climate in this region does not favor cryptococcal proliferation. This study confirmed the presence of C. neoformans and C. albidus in 20% (10/50) of randomly selected fecal samples of pigeons (Columba livia) in the Monterrey metropolitan area. The presence of this yeast in the state of Nuevo León is proof of its adaptation to the typically hot climate of the area and is consistent with recent reviews of cryptococcosis cases in several local hospitals. The two species were identified and characterized through microbiological tests and molecular identification by DNA extraction and PCR amplification of highly conserved 18S ribosomal DNA using ITS1 and ITS2 as target regions. The PCR products were sequenced and compared with those reported in GenBank.

  10. Application of microemulsion thin layer chromatography for the fingerprinting of licorice (Glycyrrhiza spp.).

    PubMed

    Cui, Shufen; Fu, Boqiang; Lee, Frank Sen-Chun; Wang, Xiaoru

    2005-12-15

    Microemulsion thin layer chromatography (ME-TLC) has been developed for the fingerprinting of aqueous extract of licorice (Glycyrrhiza spp.). The separation conditions and operational processes of the method have been optimized, and its chromatographic characteristics compared with conventional TLC. The ME-TLC system is easier to operate, and with higher resolution and better reproducibility than the conventional TLC. The separation mechanism and retention behavior of ME-TLC are found to differ significantly from conventional TLC. The technique has been applied to the analysis of different licorice species including G. uralensis, G. glabra and G. inflata; and to monitor the dynamic accumulation of active ingredients in licorice plant harvested at different times during its growing cycle in a Good Agriculture Practice (GAP) research farm. Results show that without post-chromatographic derivatization, the ME-TLC fingerprinting images of different species appear as clear, well resolved bands and with strong intensities to reveal distinctively different compositional features of the samples. The technique has also been applied successfully to monitor the dynamic accumulation of active components in licorice plant as a function of growing time in an experimental licorice farm. The study demonstrates the potential of ME-TLC technique as a rapid fingerprinting tool for the authentication and quality assessment of licorice as well as other herbs.

  11. The occurrence of Chlamydia spp. in pigs with and without clinical disease

    PubMed Central

    2012-01-01

    Background Within the genera Chlamydia, the development of refined diagnostic techniques has allowed the identification of four species that are capable of infecting pigs. The epidemiology, clinical, and zoonotic impacts of these species are however largely unknown. The study aimed to investigate the presence of Chlamydia spp. in the intestines of growing pigs and in conjunctival swabs from finisher pigs, and relate the findings to clinical signs. Results By histology, 20 of 48 pigs had intestinal lesions that may be consistent with chlamydial infection. By PCR, forty-six of the pigs were positive whereas two samples were inhibited. Sequencing of 19 DNA extracts identified these as Chlamydia suis. By immunohistochemistry, 32 of 44 samples were positive and a significant relationship was detected between macroscopically visible intestinal lesions and a high degree of infection. By real-time PCR, a significant difference was detected between pigs with and without conjunctivitis when a Ct value of 36 was employed but not when a Ct value of 38 was employed. Conclusions Chlamydia suis was demonstrated in most samples and overall, no correlation to clinical signs was detected. However, a correlation was noted between samples with a high degree of infection and the presence of clinical signs. It is possible, that the intensive pig production systems studied might predispose for the transmission and maintenance of the infection thus increasing the infectious load and the risk for disease in the pig. PMID:22280482

  12. European validation of Real-Time PCR method for detection of Salmonella spp. in pork meat.

    PubMed

    Delibato, Elisabetta; Rodriguez-Lazaro, David; Gianfranceschi, Monica; De Cesare, Alessandra; Comin, Damiano; Gattuso, Antonietta; Hernandez, Marta; Sonnessa, Michele; Pasquali, Frédérique; Sreter-Lancz, Zuzsanna; Saiz-Abajo, María-José; Pérez-De-Juan, Javier; Butrón, Javier; Prukner-Radovcic, Estella; Horvatek Tomic, Danijela; Johannessen, Gro S; Jakočiūnė, Džiuginta; Olsen, John E; Chemaly, Marianne; Le Gall, Francoise; González-García, Patricia; Lettini, Antonia Anna; Lukac, Maja; Quesne, Segolénè; Zampieron, Claudia; De Santis, Paola; Lovari, Sarah; Bertasi, Barbara; Pavoni, Enrico; Proroga, Yolande T R; Capuano, Federico; Manfreda, Gerardo; De Medici, Dario

    2014-08-01

    The classical microbiological method for detection of Salmonella spp. requires more than five days for final confirmation, and consequently there is a need for an alternative methodology for detection of this pathogen particularly in those food categories with a short shelf-life. This study presents an international (at European level) ISO 16140-based validation study of a non-proprietary Real-Time PCR-based method that can generate final results the day following sample analysis. It is based on an ISO compatible enrichment coupled to an easy and inexpensive DNA extraction and a consolidated Real-Time PCR assay. Thirteen laboratories from seven European Countries participated to this trial, and pork meat was selected as food model. The limit of detection observed was down to 10 CFU per 25 g of sample, showing excellent concordance and accordance values between samples and laboratories (100%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (100%) when the results obtained for the Real-Time PCR-based methods were compared to those of the ISO 6579:2002 standard method. The results of this international trial demonstrate that the evaluated Real-Time PCR-based method represents an excellent alternative to the ISO standard. In fact, it shows an equal and solid performance as well as it reduces dramatically the extent of the analytical process, and can be easily implemented routinely by the Competent Authorities and Food Industry laboratories.

  13. Phylogenetic Diversity and Specificity of Bacteria Closely Associated with Alexandrium spp. and Other Phytoplankton

    PubMed Central

    Jasti, Suresh; Sieracki, Michael E.; Poulton, Nicole J.; Giewat, Michael W.; Rooney-Varga, Juliette N.

    2005-01-01

    While several studies have suggested that bacterium-phytoplankton interactions have the potential to dramatically influence harmful algal bloom dynamics, little is known about how bacteria and phytoplankton communities interact at the species composition level. The objective of the current study was to determine whether there are specific associations between diverse phytoplankton and the bacteria that co-occur with them. We determined the phylogenetic diversity of bacterial assemblages associated with 10 Alexandrium strains and representatives of the major taxonomic groups of phytoplankton in the Gulf of Maine. For this analysis we chose xenic phytoplankton cultures that (i) represented a broad taxonomic range, (ii) represented a broad geographic range for Alexandrium spp. isolates, (iii) grew under similar cultivation conditions, (iv) had a minimal length of time since the original isolation, and (v) had been isolated from a vegetative phytoplankton cell. 16S rRNA gene fragments of most Bacteria were amplified from DNA extracted from cultures and were analyzed by denaturing gradient gel electrophoresis and sequencing. A greater number of bacterial species were shared by different Alexandrium cultures, regardless of the geographic origin, than by Alexandrium species and nontoxic phytoplankton from the Gulf of Maine. In particular, members of the Roseobacter clade showed a higher degree of association with Alexandrium than with other bacterial groups, and many sequences matched sequences reported to be associated with other toxic dinoflagellates. These results provide evidence for specificity in bacterium-phytoplankton associations. PMID:16000752

  14. Mimosine, a Toxin Present in Leguminous Trees (Leucaena spp.), Induces a Mimosine-Degrading Enzyme Activity in Some Rhizobium Strains

    PubMed Central

    Soedarjo, Muchdar; Hemscheidt, Thomas K.; Borthakur, Dulal

    1994-01-01

    Thirty-seven Rhizobium isolates obtained from the nodules of leguminous trees (Leucaena spp.) were selected on the basis of their ability to catabolize mimosine, a toxin found in large quantities in the seeds, foliage, and roots of plants of the genera Leucaena and Mimosa. A new medium containing mimosine as the sole source of carbon and nitrogen was used for selection. The enzymes of the mimosine catabolic pathway were inducible and were present in the soluble fraction of the cell extract of induced cells. On the basis of a comparison of the growth rates of Rhizobium strains on general carbon and nitrogen sources versus mimosine, the toxin appears to be converted mostly to biomass and carbon dioxide. Most isolates able to grow on mimosine as a source of carbon and nitrogen are also able to utilize 3-hydroxy-4-pyridone, a toxic intermediate of mimosine degradation in other organisms. PMID:16349454

  15. Molecular assays reveal the presence of Theileria spp. and Babesia spp. in Asian water buffaloes (Bubalus bubalis, Linnaeus, 1758) in the Amazon region of Brazil.

    PubMed

    Silveira, Júlia A G; de Oliveira, Cairo H S; Silvestre, Bruna T; Albernaz, Tatiana T; Leite, Rômulo C; Barbosa, José D; Oliveira, Carlos M C; Ribeiro, Múcio F B

    2016-07-01

    Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from

  16. Molecular assays reveal the presence of Theileria spp. and Babesia spp. in Asian water buffaloes (Bubalus bubalis, Linnaeus, 1758) in the Amazon region of Brazil.

    PubMed

    Silveira, Júlia A G; de Oliveira, Cairo H S; Silvestre, Bruna T; Albernaz, Tatiana T; Leite, Rômulo C; Barbosa, José D; Oliveira, Carlos M C; Ribeiro, Múcio F B

    2016-07-01

    Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from

  17. Molecular detection of Theileria spp. and Babesia spp. in sheep and ixodid ticks from the northeast of Iran.

    PubMed

    Razmi, Gholamreza; Pourhosseini, Moslem; Yaghfouri, Saeed; Rashidi, Ahmad; Seidabadi, Mohsen

    2013-02-01

    Theilerioses and babesioses are important diseases in Iranian sheep. The present study was undertaken to identify and classify/specify Theileria spp. and Babesia spp. in sheep and vector ticks. Investigation was carried out from 2009 to 2011 in the Khorasan Razavi Province, Iran. In total, 302 sheep originating from 60 different flocks were clinically examined and their blood collected. In addition, from the same flocks, ixodid ticks were sampled. Stained blood smears were microscopically examined for the presence of Theileria and Babesia organisms, and a semi-nested PCR was used for subsequent molecular specification. From the ticks, salivary glands and uterus were isolated and subsequently analyzed by semi-nested PCR. Piroplasm organisms were observed in 29% of the blood smears with low parasitemia, whereas 65% of the blood samples yielded positive PCR findings. The presence of Theileria ovis (55.6%), Theileria lestoquardi, and mixed infection with Theileria spp. and Babesia ovis were detected by semi-nested PCR in 0.3%, 5.6%, and 0.99%, respectively. In total, 429 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 376; 87.6% of the total), followed by Hyalomma marginatum turanicum (n = 30; 7.0%), Dermacentor raskemensis (n = 12; 2.8%), Hyalomma anatolicum anatolicum (n = 7; 1.6%), Dermacentor marginatus (n = 2; 0.5%), Rhipicephalus bursa (n = 1; 0.2%), and Haemaphysalis sp. (n = 1; 0.2%). Of the positive R. turanicus samples, 5 (5.7%) were infected with T. ovis and 2 (2.9%) with T. lestoquardi. Neither Babesia ovis nor Babesia motasi infection was detected in salivary glands or uterine samples of the ticks. The results also suggest that R. turanicus could be the vector responsible for transmission of the 2 Theileria species.

  18. Detection of Alicyclobacillus spp. in Fruit Juice by Combination of Immunomagnetic Separation and a SYBR Green I Real-Time PCR Assay

    PubMed Central

    Yuan, Yahong; Liu, Bin; Wang, Ling; Yue, Tianli

    2015-01-01

    An approach based on immunomagnetic separation (IMS) and SYBR Green I real-time PCR (real-time PCR) with species-specific primers and melting curve analysis was proposed as a rapid and effective method for detecting Alicyclobacillus spp. in fruit juices. Specific primers targeting the 16S rDNA sequences of Alicyclobacillus spp. were designed and then confirmed by the amplification of DNA extracted from standard strains and isolates. Spiked samples containing known amounts of target bacteria were used to obtain standard curves; the correlation coefficient was greater than 0.986 and the real-time PCR amplification efficiencies were 98.9%- 101.8%. The detection limit of the testing system was 2.8×101 CFU/mL. The coefficient of variation for intra-assay and inter-assay variability were all within the acceptable limit of 5%. Besides, the performance of the IMS-real-time PCR assay was further investigated by detecting naturally contaminated kiwi fruit juice; the sensitivity, specificity and accuracy were 91.7%, 95.9% and 95.3%, respectively. The established IMS-real-time PCR procedure provides a new method for identification and quantitative detection of Alicyclobacillus spp. in fruit juice. PMID:26488469

  19. Partial identification of antifungal compounds from Punica granatum peel extracts.

    PubMed

    Glazer, Ira; Masaphy, Segula; Marciano, Prosper; Bar-Ilan, Igal; Holland, Doron; Kerem, Zohar; Amir, Rachel

    2012-05-16

    Aqueous extracts of pomegranate peels were assayed in vitro for their antifungal activity against six rot fungi that cause fruit and vegetable decay during storage. The growth rates of Alternaria alternata , Stemphylium botryosum , and Fusarium spp. were significantly inhibited by the extracts. The growth rates were negatively correlated with the levels of total polyphenolic compounds in the extract and particularly with punicalagins, the major ellagitannins in pomegranate peels. Ellagitannins were also found to be the main compounds in the bioactive fractions using bioautograms, and punicalagins were identified as the main bioactive compounds using chromatographic separation. These results suggest that ellagitannins, and more specifically punicalagins, which are the dominant compounds in pomegranate peels, may be used as a control agent of storage diseases and to reduce the use of synthetic fungicides. PMID:22533815

  20. The non-competitive blockade of GABAA receptors by an aqueous extract of water hemlock (Cicuta douglasii) tubers.

    PubMed

    Green, Benedict T; Goulart, Camila; Welch, Kevin D; Pfister, James A; McCollum, Isabelle; Gardner, Dale R

    2015-12-15

    Water hemlocks (Cicuta spp.) are acutely toxic members of the Umbellierae family; the toxicity is due to the presence of C17-polyacetylenes such as cicutoxin. There is only limited evidence of noncompetitive antagonism by C17-polyacetylenes at GABAA receptors. In this work with WSS-1 cells, we documented the noncompetitive blockade of GABAA receptors by an aqueous extract of water hemlock (Cicuta douglasii) and modulated the actions of the extract with a pretreatment of 10 μM midazolam. PMID:26415905

  1. Quantitative Microbial Risk Assessment for Campylobacter spp. on Ham in Korea.

    PubMed

    Lee, Jeeyeon; Ha, Jimyeong; Kim, Sejeong; Lee, Heeyoung; Lee, Soomin; Yoon, Yohan

    2015-01-01

    The objective of this study was to evaluate the risk of illness from Campylobacter spp. on ham. To identify the hazards of Campylobacter spp. on ham, the general characteristics and microbial criteria for Campylobacter spp., and campylobacteriosis outbreaks were investigated. In the exposure assessment, the prevalence of Campylobacter spp. on ham was evaluated, and the probabilistic distributions for the temperature of ham surfaces in retail markets and home refrigerators were prepared. In addition, the raw data from the Korea National Health and Nutrition Examination Survey (KNHNES) 2012 were used to estimate the consumption amount and frequency of ham. In the hazard characterization, the Beta-Poisson model for Campylobacter spp. infection was used. For risk characterization, a simulation model was developed using the collected data, and the risk of Campylobacter spp. on ham was estimated with @RISK. The Campylobacter spp. cell counts on ham samples were below the detection limit (<0.70 Log CFU/g). The daily consumption of ham was 23.93 g per person, and the consumption frequency was 11.57%. The simulated mean value of the initial contamination level of Campylobacter spp. on ham was -3.95 Log CFU/g, and the mean value of ham for probable risk per person per day was 2.20×10(-12). It is considered that the risk of foodborne illness for Campylobacter spp. was low. Furthermore, these results indicate that the microbial risk assessment of Campylobacter spp. in this study should be useful in providing scientific evidence to set up the criteria of Campylobacter spp.. PMID:26761897

  2. Toxocara spp. seroprevalence in sheep from southern Brazil.

    PubMed

    Rassier, Gabriela Lopes; Borsuk, Sibele; Pappen, Felipe; Scaini, Carlos Jaime; Gallina, Tiago; Villela, Marcos Marreiro; da Rosa Farias, Nara Amélia; Benavides, Magda Vieira; Berne, Maria Elisabeth Aires

    2013-09-01

    Visceral toxocariasis is a neglected parasitic zoonosis that occurs through the ingestion of embryonated Toxocara spp. eggs. A wide range of animal species can act as paratenic hosts for this ascarid. The main risk factor for humans is the ingestion of the eggs from contaminated soil; however, infection can also occur through the ingestion of contaminated raw or undercooked infected meat from paratenic hosts. The aim of this study was to verify the presence of Toxocara spp.-specific antibodies in sheep and to determine the risk factors associated with the infection of sheep in Rio Grande do Sul (a major sheep-producing and sheep-consuming state) in southern Brazil. Serum samples collected from 1,642 sheep were tested using an IgG enzyme-linked immunosorbent assay based on the excretory-secretory Toxocara canis antigen. Seroprevalence was 29.0% (477/1,642), and every farm included in the study contained at least one seropositive animal. These results indicate that T. canis infection is widely distributed among sheep herds in Rio Grande do Sul and that it represents a potential risk to human health. PMID:23832639

  3. Detection of Acinetobacter spp. in rural drinking water supplies.

    PubMed Central

    Bifulco, J M; Shirey, J J; Bissonnette, G K

    1989-01-01

    A bacteriological survey was conducted of untreated, individual groundwater supplies in Preston County, W.Va. Nearly 60% of the water supplies contained total coliforms in excess of the U.S. Environmental Protection Agency maximum contaminant level of 1 CFU/100 ml. Approximately one-third of the water systems contained fecal coliforms and/or fecal streptococci. Acinetobacter spp. were detected in 38% of the groundwater supplies at an arithmetic mean density of 8 CFU/100 ml and were present in 16% of the water supplies in the absence of total coliforms, posing some concern about the usefulness of total coliforms as indicators of the presence of this opportunistic pathogen. Slime production, a virulence factor for A. calcoaceticus, was not significantly different between well water isolates and clinical strains, suggesting some degree of pathogenic potential for strains isolated from groundwater. In addition, several Acinetobacter isolates were able to interfere with sheen production by some coliform bacteria on M-Endo medium, adding further to the possible significance of Acinetobacter spp. in groundwater supplies. PMID:2529816

  4. Virulence of Meloidogyne spp. and Induced Resistance in Grape Rootstocks

    PubMed Central

    McKenry, Michael V.; Anwar, Safdar A.

    2007-01-01

    Harmony grape rootstock displays resistance to several Meloidogyne spp. but that resistance is not durable in commercial vineyard settings. A 2-year experiment in a microplot setting revealed host specificities of two virulent populations of Meloidogyne arenaria and an avirulent population of Meloidogyne incognita. In a subsequent split-root experiment, the avirulent nematode population was demonstrated to induce resistance to the virulent nematode population. To quantify the level of resistance, reproduction of the virulent nematode population was determined 63 days after being challenged by an avirulent nematode population using a range of inoculum densities and timeframes. Induction of resistance became apparent when the virulent nematode population was inoculated 7 days after the avirulent nematode population and increased thereafter. The level of induced resistance increased with increased inoculum levels of the avirulent nematode population. Root systems of perennial crops are commonly fed upon simultaneously by multiple nematode species. These two studies indicate that field populations can become preferentially virulent upon one or multiple rootstocks and that co-inhabiting populations may induce existing resistance mechanisms. In perennial crops, it is common for numerous nematode species besides Meloidogyne spp. to be present, including some that feed without causing apparent damage. PMID:19259475

  5. Detection of Paracoccidioides spp. in environmental aerosol samples.

    PubMed

    Arantes, Thales Domingos; Theodoro, Raquel Cordeiro; Da Graça Macoris, Severino Assis; Bagagli, Eduardo

    2013-01-01

    Taking into account that paracoccidioidomycosis infection occurs by inhalation of the asexual conidia produced by Paracoccidioides spp. in its saprobic phase, this work presents the collection of aerosol samples as an option for environmental detection of this pathogen, by positioning a cyclonic air sampler at the entrance of armadillo burrows. Methods included direct culture, extinction technique culture and Nested PCR of the rRNA coding sequence, comprising the ITS1-5.8S-ITS2 region. In addition, we evaluated one armadillo (Dasypus novemcinctus) as a positive control for the studied area. Although the pathogen could not be isolated by the culturing strategies, the aerosol sampling associated with molecular detection through Nested PCR proved the best method for discovering Paracoccidioides spp. in the environment. Most of the ITS sequences obtained in this investigation proved to be highly similar with the homologous sequences of Paracoccidioides lutzii from the GenBank database, suggesting that this Paracoccidioides species may not be exclusive to mid-western Brazil as proposed so far. PMID:22762209

  6. Identification of Staphylococcus spp. using (GTG)₅-PCR fingerprinting.

    PubMed

    Svec, Pavel; Pantůček, Roman; Petráš, Petr; Sedláček, Ivo; Nováková, Dana

    2010-12-01

    A group of 212 type and reference strains deposited in the Czech Collection of Microorganisms (Brno, Czech Republic) and covering 41 Staphylococcus species comprising 21 subspecies was characterised using rep-PCR fingerprinting with the (GTG)₅ primer in order to evaluate this method for identification of staphylococci. All strains were typeable using the (GTG)₅ primer and generated PCR products ranging from 200 to 4500 bp. Numerical analysis of the obtained fingerprints revealed (sub)species-specific clustering corresponding with the taxonomic position of analysed strains. Taxonomic position of selected strains representing the (sub)species that were distributed over multiple rep-PCR clusters was verified and confirmed by the partial rpoB gene sequencing. Staphylococcus caprae, Staphylococcus equorum, Staphylococcus sciuri, Staphylococcus piscifermentans, Staphylococcus xylosus, and Staphylococcus saprophyticus revealed heterogeneous fingerprints and each (sub)species was distributed over several clusters. However, representatives of the remaining Staphylococcus spp. were clearly separated in single (sub)species-specific clusters. These results showed rep-PCR with the (GTG)₅ primer as a fast and reliable method applicable for differentiation and straightforward identification of majority of Staphylococcus spp.

  7. Type IV secretion system of Brucella spp. and its effectors

    PubMed Central

    Ke, Yuehua; Wang, Yufei; Li, Wengfeng; Chen, Zeliang

    2015-01-01

    Brucella spp. are intracellular bacterial pathogens that cause infection in domestic and wild animals. They are often used as model organisms to study intracellular bacterial infections. Brucella VirB T4SS is a key virulence factor that plays important roles in mediating intracellular survival and manipulating host immune response to infection. In this review, we discuss the roles of Brucella VirB T4SS and 15 effectors that are proposed to be crucial for Brucella pathogenesis. VirB T4SS regulates the inflammation response and manipulates vesicle trafficking inside host cells. VirB T4SS also plays crucial roles in the inhibition of the host immune response and intracellular survival during infection. Here, we list the key molecular events in the intracellular life cycle of Brucella that are potentially targeted by the VirB T4SS effectors. Elucidating the functions of these effectors will help clarify the molecular role of T4SS during infection. Furthermore, studying the effectors secreted by Brucella spp. might provide insights into the mechanisms used by the bacteria to hijack the host signaling pathways and aid in the development of better vaccines and therapies against brucellosis. PMID:26528442

  8. Detection of Paracoccidioides spp. in environmental aerosol samples.

    PubMed

    Arantes, Thales Domingos; Theodoro, Raquel Cordeiro; Da Graça Macoris, Severino Assis; Bagagli, Eduardo

    2013-01-01

    Taking into account that paracoccidioidomycosis infection occurs by inhalation of the asexual conidia produced by Paracoccidioides spp. in its saprobic phase, this work presents the collection of aerosol samples as an option for environmental detection of this pathogen, by positioning a cyclonic air sampler at the entrance of armadillo burrows. Methods included direct culture, extinction technique culture and Nested PCR of the rRNA coding sequence, comprising the ITS1-5.8S-ITS2 region. In addition, we evaluated one armadillo (Dasypus novemcinctus) as a positive control for the studied area. Although the pathogen could not be isolated by the culturing strategies, the aerosol sampling associated with molecular detection through Nested PCR proved the best method for discovering Paracoccidioides spp. in the environment. Most of the ITS sequences obtained in this investigation proved to be highly similar with the homologous sequences of Paracoccidioides lutzii from the GenBank database, suggesting that this Paracoccidioides species may not be exclusive to mid-western Brazil as proposed so far.

  9. Mangrove Ecosystems: An Adopted Habitat for Pathogenic Salmonella spp.

    PubMed

    Poharkar, Krupali V; Kerkar, Savita; D'Costa, Dilecta; Doijad, Swapnil; Barbuddhe, S B

    2016-03-01

    Mangroves are affected by industrial and anthropogenic factors. Although mangroves have been widely studied, investigations of pathogens that may affect public health significance are largely lacking even while incidences of diseases linked with the consumption of mangrove-associated food have increased. A total of 150 samples of water, sediment, and biota were collected from ten mangrove ecosystems in Goa, India. Total viable counts of pathogens such as E. coli, Listeria, Salmonella, and Vibrio spp. ranged from 1.25 to 3.9 × 10(3) cfu/ mL, which were above the relevant standards. Salmonella counts were the highest at 3.1 to 3.9 × 10(3)cfu/mL, with a prevalence of 40%. Considering its high prevalence, the virulence of Salmonella spp. was studied. The invA gene was detected in 35% of the Salmonella isolates by polymerase chain reaction (PCR). The findings suggested that pathogens adapt to this habitat, resulting in contamination of the indigenous fauna. PMID:26931537

  10. Frequency of Antibiotic-Producing Pseudomonas spp. in Natural Environments

    PubMed Central

    Raaijmakers, J. M.; Weller, D. M.; Thomashow, L. S.

    1997-01-01

    The antibiotics phenazine-1-carboxylic acid (PCA) and 2,4-diacetylphloroglucinol (Phl) are major determinants of biological control of soilborne plant pathogens by various strains of fluorescent Pseudomonas spp. In this study, we described primers and probes that enable specific and efficient detection of a wide variety of fluorescent Pseudomonas strains that produce various phenazine antibiotics or Phl. PCR analysis and Southern hybridization demonstrated that specific genes within the biosynthetic loci for Phl and PCA are conserved among various Pseudomonas strains of worldwide origin. The frequency of Phl- and PCA-producing fluorescent pseudomonads was determined on roots of wheat grown in three soils suppressive to take-all disease of wheat and four soils conducive to take-all by colony hybridization followed by PCR. Phenazine-producing strains were not detected on roots from any of the soils. However, Phl-producing fluorescent pseudomonads were isolated from all three take-all-suppressive soils at densities ranging from approximately 5 x 10(sup5) to 2 x 10(sup6) CFU per g of root. In the complementary conducive soils, Phl-producing pseudomonads were not detected or were detected at densities at least 40-fold lower than those in the suppressive soils. We speculate that fluorescent Pseudomonas spp. that produce Phl play an important role in the natural suppressiveness of these soils to take-all disease of wheat. PMID:16535555

  11. [Dendrolimus spp. damage monitoring by using NOAA/AVHRR data].

    PubMed

    Zhang, Yushu; Ban, Xianxiu; Chen, Pengshi; Feng, Rui; Ji, Ruipeng; Xiao, Yan

    2005-05-01

    This paper approached the feasibility of quantitatively monitoring Dendrolimus spp. damage by using NOAA/ AVHRR data. The damaged rate of needle leaf was used to represent Dendrolimus spp. harming degree, and < 30%, 30%-60% and > 60% of damaged rate was defined as low, medium and severe harming degree, respectively. The correlation equation of damaged rate and normalized vegetation index (NDVI) was established, based on the ground spectrum observation. The NDVI was 0.8823 when no damage occurred. A relative NDVI value of damaged to undamaged area was used to express the remote sensing index of low, medium and severe harming degree. The index was 1 for undamaged forest, and 0.78-1, 0.57-0.78 and < 0.57 for low, medium and severe harming degrees, respectively. The mixed pixels were separated by linear addable vertical vegetation index in the monitoring, and the quantitative monitoring and analysis was accomplished for years when the three damage degrees happened. It was shown that AVHRR data could be more available in quantitatively monitoring and analyzing serious damage, while low degree damage was difficult to distinguish by AVHRR data, due to the differences of surface properties and atmospheric influences, as well as the lower space resolution of NOAA/AVHRR. The damaged area estimated by AVHRR was 12.1%-14.3% lower than that by TM.

  12. Gasterophilus spp. infections in horses from northern and central Kazakhstan.

    PubMed

    Ibrayev, Baltabek; Lider, Lyudmila; Bauer, Christian

    2015-01-15

    A cross-sectional survey was performed to obtain current data on the gastrointestinal myiasis of horses in the provinces of Kostanay, Akmola and Karagandy, northern and central Kazakhstan. The stomach, small intestine and rectum of 148 slaughter horses were examined for Gasterophilus spp. larvae during a 26-month study period. All horses were infected with 2nd and 3rd stage larvae (mean intensity: 803±350), and 22% of them harboured >1000 Gasterophilus spp. larvae each. Four species were identified: G. intestinalis (prevalence: 100%; mean intensity: 361±240 larvae), G. haemorrhoidalis (100%; 353±191), G. nasalis (100%; 73±36) and G. pecorum (91.2%; 18±10). Horses aged<2 years were higher infected with Gasterophilus larvae than 2-4 years old animals. Both the prevalence and extremely high intensity of Gasterophilus infections of horses in these Kazakh regions suggest respective control measurements to improve the health and performance of the animals and to increase the economic income of horse owners. PMID:25522954

  13. Temporal study of Nosema spp. in a cold climate.

    PubMed

    Forsgren, Eva; Fries, Ingemar

    2013-02-01

    In a nationwide Swedish survey, 967 honey bee colonies from 521 beekeepers were sampled in the spring of 2007 and the samples assayed for Nosema spp. infections. Of the 319 positive samples, only 32 samples contained a proportion of N. ceranae DNA in mixed infections with both Nosema spp. above the cut-off point chosen for comparisons of 1%. Only one pure N. ceranae infection was found, with the rest 284 infected samples assayed being pure N. apis infections. In 2009 and 2011, beekeepers or bee inspectors providing N. ceranae mixed positive bee samples in 2007 were again asked to submit samples (2009, n = 96; 2011, n = 83). No trend of an increased proportion of N. ceranae-infected samples could be found. The proportion of N. ceranae DNA in samples with mixed infection did not increase between 2007 and 2011. It is concluded that N. apis is still the dominating Microsporidia infection in honey bees in Sweden and that there is no tendency for one species replacing the other.

  14. Bartonella spp. in cats from Buenos Aires, Argentina.

    PubMed

    Cicuttin, Gabriel L; Brambati, Diego F; De Gennaro, María F; Carmona, Fernando; Isturiz, María L; Pujol, Laura E; Belerenian, Guillermo C; Gil, Horacio

    2014-01-10

    In Argentina, data on the presence of members of the genus Bartonella is scarce. To increase knowledge about these zoonotic pathogens in this country, the presence and variability of Bartonella spp. was investigated in cats and dogs from Buenos Aires. Bartonella spp. was detected in 17.8% of cats, while all dogs tested negative by PCR and Reverse Line Blot. B. henselae was the most frequent species, being detected in 11.9% (14/101), while B. clarridgeiae was found in only 5.9% (6/101) of the cats. Afterwards, B. henselae isolates and positive blood samples were characterized by Multiple Locus Sequence Typing (MLST) and Multiple Locus Variable Number Tandem Repeats Analysis (MLVA). As result, four different MLST sequence types (ST) and eight MLVA profiles were identified. ST 1 was the most frequent variant found in cats, followed by ST 8. Interestingly, some of the MLVA profiles that were detected in this study have been previously associated with human disease, and represents a potential risk of infection. Veterinarians and physicians should consider the presence of these emerging pathogens in their diagnostic routine.

  15. Molecular Characterization of Cryptosporidium spp. in Children from Mexico

    PubMed Central

    Valenzuela, Olivia; González-Díaz, Mariana; Garibay-Escobar, Adriana; Burgara-Estrella, Alexel; Cano, Manuel; Durazo, María; Bernal, Rosa M.; Hernandez, Jesús; Xiao, Lihua

    2014-01-01

    Cryptosporidiosis is a parasitic disease caused by Cryptosporidium spp. In immunocompetent individuals, it usually causes an acute and self-limited diarrhea; in infants, infection with Cryptosporidium spp. can cause malnutrition and growth retardation, and declined cognitive ability. In this study, we described for the first time the distribution of C. parvum and C. hominis subtypes in 12 children in Mexico by sequence characterization of the 60-kDa glycoprotein (GP60) gene of Cryptosporidium. Altogether, 7 subtypes belonging to 4 subtype families of C. hominis (Ia, Ib, Id and Ie) and 1 subtype family of C. parvum (IIa) were detected, including IaA14R3, IaA15R3, IbA10G2, IdA17, IeA11G3T3, IIaA15G2R1 and IIaA16G1R1. The frequency of the subtype families and subtypes in the samples analyzed in this study differed from what was observed in other countries. PMID:24755606

  16. Presence of microorganisms from isolated Megaselia spp. in foodservice establishments.

    PubMed

    Soler, Carla; Esteban, J Guillermo; Jiménez, Ricardo; Mañes, Jordi; Soriano, José Miguel

    2015-06-01

    Introducción: la transmisión de patógenos por insectos es una creciente preocupación para la salud pública. Más concretamente, las moscas son conocidas por ser capaces de transmitir el agente infeccioso mecánicamente. Objetivo: el presente trabajo muestra un estudio en los servicios de restauración en los que se aisló por primera vez en la literatura Megaselia spp, detectándose la presencia de microorganismos en estas moscas. Método: se basa en análisis microbiológicos y entomológicos. Resultados: la presencia de aerobios mesófilos y Enterobacteriaceae se han encontrado en todas las muestras, superando los límites establecidos en el 41,7% (5/12) para las bacterias aerobias mesófilas y el 66,7% (8/12) para Enterobacteriaceae. Por otra parte, en el 25 y 66,7% de las moscas analizadas se detectó la presencia de Escherichia coli y Staphylococcus aureus, respectivamente. Conclusiones: hay un binomio entre la presencia de microorganismos y Megaselia spp., lo que demuestra la importancia de mantener una vigilancia más estricta en las medidas higiénico-sanitarias en los servicios de restauración.

  17. Mangrove Ecosystems: An Adopted Habitat for Pathogenic Salmonella spp.

    PubMed

    Poharkar, Krupali V; Kerkar, Savita; D'Costa, Dilecta; Doijad, Swapnil; Barbuddhe, S B

    2016-03-01

    Mangroves are affected by industrial and anthropogenic factors. Although mangroves have been widely studied, investigations of pathogens that may affect public health significance are largely lacking even while incidences of diseases linked with the consumption of mangrove-associated food have increased. A total of 150 samples of water, sediment, and biota were collected from ten mangrove ecosystems in Goa, India. Total viable counts of pathogens such as E. coli, Listeria, Salmonella, and Vibrio spp. ranged from 1.25 to 3.9 × 10(3) cfu/ mL, which were above the relevant standards. Salmonella counts were the highest at 3.1 to 3.9 × 10(3)cfu/mL, with a prevalence of 40%. Considering its high prevalence, the virulence of Salmonella spp. was studied. The invA gene was detected in 35% of the Salmonella isolates by polymerase chain reaction (PCR). The findings suggested that pathogens adapt to this habitat, resulting in contamination of the indigenous fauna.

  18. Biosurfactant production in sugar beet molasses by some Pseudomonas spp.

    PubMed

    Onbasli, Dilsad; Aslim, Belma

    2009-01-01

    In this study rhamnolipid biosurfactant production was investigated in eighteen strains of Pseudomonas spp.. Rhamnolipid by these strains was determined by a spectrophotometric method in nutrient broth medium (NB). From the 18 strains screened, two Pseudomonas strains (Pseudomonas luteola B17 and Pseudomonas putida B12) which had produced the highest percentage yield of rhamnolipid were examined for rhamnolipid production at different incubation times (24, 48 and 72 hr) and different sugar beet molasses concentrations [1-5% w/v concentration (1-5 g molasses/100 ml water)]. The rhamnolipid production increased with the increase in the concentration of molasses and maximum production occurred when 5 % (w/v) of molasses were used. At the same time, maximum rhamnolipid production occurred after 72 hr of incubation. When the amount of rhamnolipid produced at different incubation times (24, 48 and 72 hr) and with different concentrations of molasses [1-5 % w/v concentration (1-5 g molasses/100 ml water)] by Pseudomonas spp.; was compared, no significant difference in amount of production was seen. These studies show that the waste product from sugar industry may be suggested for important biotechnological processes such as rhamnolipid production.

  19. Cytotoxic effect of acriflavine against clinical isolates of Acanthamoeba spp.

    PubMed

    Polat, Zubeyda Akin; Karakus, Gulderen

    2013-02-01

    Acanthamoeba keratitis (AK) is a potentially devastating and sight-threatening infection of the cornea caused by the ubiquitous free-living amoebae, Acanthamoeba species. Its eradication is difficult because the amoebas encyst, making it highly resistant to anti-amoebic drugs. Acriflavine neutral (ACF) has been used for treatment of microbial infections for humans and fishes. The aim of our study was to evaluate the time-dependent cytotoxicities of ACF against Acanthamoeba spp. Trophozoites and cysts of three different strains (strain PAT06 Acanthamoeba castellanii, strain 2HH Acanthamoeba hatchetti, and strain 11DS A. hatchetti) of Acanthamoeba spp. were tested. All strains had been isolated from patients suffering from a severe AK. The effects of the ACF with the concentrations ranging from 15 to 500 mg mL(-1) on the cytotoxicity of Acanthamoeba strains were examined. ACF showed a time- and dose-dependent amebicidal action on the trophozoites and cysts. Pat06 (A. castellanii) was the most resistant, while strain 11DS (A. hatchetti) was the most sensitive. As a result, ACF could be concluded as a new agent for the treatment of Acanthamoeba infections. On the other hand, it still needs to be further evaluated by in vivo test systems to confirm the efficiency of its biological effect. PMID:23052789

  20. Efficacy of potato seeds disinfection products to control Erwinia spp.

    PubMed

    Dupuis, B; Garcia, N; Boels, G

    2008-01-01

    Erwinia spp. provokes soft rot on potato tubers during storage. No disinfection products are available on the market in the European Union to control these bacteria. We tested 3 products presented as good candidates to cure potato tubers from bacterial diseases. First, Anthium 500 (Du Pont de Nemours) a product based on chlorine dioxyde, then Phostrol (Nufarm) with phosphoric acid as a.i. and finally Solucuivre (Proval), a copper based product. We firstly managed disinfection trials: high Erwinia contaminated potato seed samples were treated by immersion and were then incubated, we observed the percentage of tubers rotting. Secondly, we managed protection trials: protected healthy tubers were incubated during 23 days in contact with rotting tubers. We evaluated weight loss after symptoms development. No tested product was effective to control Erwinia spp. on seed tubers in our trials conditions. Furthermore, we observed more rot development after Phostrol and Solucuivre application. We suppose that the product couldn't reach the latent bacteria and weakened the tubers. No protection effect was observed.