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Discrimination between human, pig and ruminant fecal contaminations in a river catchment by real-time PCR using host-specific markers ArchiMer
Mieszkin, Sophie; Furet, Jean-pierre; Corthier, Gerard; Pommepuy, Monique; Le Saux, Jean-claude; Bougeard, Morgane; Hervio Heath, Dominique; Gourmelon, Michele.
The microbiological quality of coastal waters and shellfish harvesting areas in Brittany (France) can be affected by faecal pollutions from human activities and animal breeding (especially pigs and cattle). To discriminate among faecal pollution of human and animal origin, a library-independent microbial source tracking method was selected: Bacteroidales host-specific 16S rRNA gene markers by real-time Polymerase Chain Reaction (PCR). A human-specific Bacteroidales marker (Hum-1-Bac) was designed. Tested on faecal samples, the Hum-1-Bac marker showed 95 % sensitivity and 95 % specificity (n= 80). Average values (± STD) of the Hum-1-Bac marker were found to be 7.3 ± 1.4 16S rRNA gene copies per g wet faeces in human faeces samples (n=10) and 5.7 ± 1.3 log10...
Tipo: Text Palavras-chave: Microbial Source Tracking; Host-Specific Bacteroidales Marker; 16S rRNA Gene; Real-Time PCR; Faecal Contamination.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/00021/13259/10306.pdf
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Phylogenetic analysis of Bacteroidales 16S rRNA gene sequences from human and animal effluents and assessment of ruminant faecal pollution by real-time PCR ArchiMer
Mieszkin, Sophie; Yala, Jean Fabrice; Joubrel, Rozenn; Gourmelon, Michele.
Aims: The aims of this study were to evaluate the host-specific distribution of Bacteroidales 16S rRNA gene sequences from human- and animal-related effluents and faeces, and to define a ruminant-specific marker. Methods and Results: Bacteroidales 16S rRNA gene clone libraries were constructed from samples of effluent (sewage, bovine manure and pig slurry) and faeces (human, bovine, pig and wild bird), using PCR primers targeting order Bacteroidales. The phylogenetic analysis revealed six main distinct human-, bovine-, pig- and wild bird-specific clusters. From the bovine-specific cluster II, we designed a ruminant-specific marker, Rum-2-Bac, and this showed 97% sensitivity (n = 30) and 100% specificity (n = 40) when tested by TaqMan (R) real-time PCR....
Tipo: Text Palavras-chave: 16S rRNA gene; Bacteroidales; Faecal contamination; Microbial Source Tracking; Sewage and manure.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00000/11156/8235.pdf
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Origin of fecal contamination in waters from contrasted areas: Stanols as Microbial Source Tracking markers ArchiMer
Derrien, M.; Jarde, E.; Gruau, G.; Pourcher, A. M.; Gourmelon, Michele; Jadas-hecart, Alain; Wickmann, A. C. Pierson.
Improving the microbiological quality of coastal and river waters relies on the development of reliable markers that are capable of determining sources of fecal pollution. Recently, a principal component analysis (PCA) method based on six stanol compounds (i.e. 5 beta-cholestan-3 beta-ol (coprostanol), 5 beta-cholestan-3 alpha-ol (epicoprostanol), 24-methyl-5 alpha-cholestan-3 beta-ol (campestanol), 24-ethyl-5 alpha-cholestan-3 beta-ol (sitostanol), 24-ethyl-5 beta-cholestan-3 beta-ol (24-ethylcoprostanol) and 24-ethyl-5 beta-cholestan-3 alpha-ol (24-ethylepicoprostanol)) was shown to be suitable for distinguishing between porcine and bovine feces. In this study, we tested if this PCA method, using the above six stanols, could be used as a tool in...
Tipo: Text Palavras-chave: Stanols; Microbial Source Tracking; Principal component analysis; Microbial markers.
Ano: 2012 URL: http://archimer.ifremer.fr/doc/00090/20133/17829.pdf
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