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Comparative assessment of human and farm animal faecal microbiota using real-time quantitative PCR ArchiMer
Furet, Jean-pierre; Firmesse, Olivier; Gourmelon, Michele; Bridonneau, Chantal; Tap, Julien; Mondot, Stanislas; Dore, Joel; Corthier, Gerard.
Pollution of the environment by human and animal faecal pollution affects the safety of shellfish, drinking water and recreational beaches. To pinpoint the origin of contaminations, it is essential to define the differences between human microbiota and that of farm animals. A strategy based on real-time quantitative PCR (qPCR) assays was therefore developed and applied to compare the composition of intestinal microbiota of these two groups. Primers were designed to quantify the 16S rRNA gene from dominant and subdominant bacterial groups. TaqMan((R)) probes were defined for the qPCR technique used for dominant microbiota. Human faecal microbiota was compared with that of farm animals using faecal samples collected from rabbits, goats, horses, pigs, sheep...
Tipo: Text Palavras-chave: Quantitative PCR; Faecal microbiota; Human; Farm animals.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/00000/11071/11297.pdf
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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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Estimation of Pig Fecal Contamination in a River Catchment by Real-Time PCR Using Two Pig-Specific Bacteroidales 16S rRNA Genetic Markers ArchiMer
Mieszkin, Sophie; Furet, Jean-pierre; Corthier, Gerard; Gourmelon, Michele.
The microbiological quality of coastal or river water can be affected by fecal contamination from human or animal sources. To discriminate pig fecal pollution from other pollution, a library-independent microbial source tracking method targeting Bacteroidales host-specific 16S rRNA gene markers by real-time PCR was designed. Two pig-specific Bacteroidales markers (Pig-1-Bac and Pig-2-Bac) were designed using 16S rRNA gene Bacteroidales clone libraries from pig feces and slurry. For these two pig markers, 98 to 100% sensitivity and 100% specificity were obtained when tested by TaqMan real-time PCR. A decrease in the concentrations of Pig-1-Bac and Pig-2-Bac markers was observed throughout the slurry treatment chain. The two newly designed pig-specific...
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Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6493.pdf
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