Capillary electrophoresis single-strand conformation polymorphism for the monitoring of gastrointestinal microbiota of chicken flocks.
The objective of the present study was to evaluate the capillary electrophoresis single-strand conformation polymorphism (CE-SSCP) to characterize poultry gut microbiota and the ability of this molecular method to detect modifications related to rearing conditions to be used as an epidemiological tool. The V3 region of the 16S rRNA gene was selected as the PCR target. Our results showed that this method provides reproducible data. The microbiota analysis of individuals showed that variability between individual fingerprints was higher for ileum and cloaca than for ceca. However, pooling the samples decreased this variability. To estimate the variability within and between farms, we compared molecular gut patterns of animals from the same hatchery reared under similar conditions and fed the same diet in 2 separate farms. Total aerobic bacteria, coliforms, and lactic acid bacteria were enumerated using conventional bacteriological methods. A significant difference was observed for coliforms present in the ceca and the cloaca depending on the farm. Ileal contents fingerprints were more closely related to those of cloacal contents than to those of ceca contents. When comparing samples from the 2 farms, a specific microbiota was highlighted for each farm. For each gut compartment, the microbiota fingerprints were joined in clusters according to the farm. Thus, this rapid and potentially high-throughput method to obtain gut flora fingerprints is sensitive enough to detect a "farm effect" on the balance of poultry gut microbiota despite the birds being fed the same regimens and reared under similar conditions.
Anses, Laboratoire de Ploufragan-Plouzané, Unité Hygiène et Qualité des Produits Avicoles et Porcins, BP53, 22440 Ploufragan, France. christine.pisavin@univ-rennes1.fr
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Pissavin
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Burel
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Gabriel
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Beven
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Mallet
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Maurice
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Queguiner
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Lessire
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Fravalo
D000822:Animal Husbandry; D000818:Animals; D001419:Bacteria / Q000145:classification*; D002645:Chickens / Q000382:microbiology*; D004269:DNA, Bacterial; D019075:Electrophoresis, Capillary / Q000379:methods*; D041981:Gastrointestinal Tract / Q000382:microbiology*; D008297:Male; D011110:Polymorphism, Genetic
D004269:DNA, Bacterial