Assessment of a molecular tool (CE-SSCP) to study balance of caecal flora of SPF piglet's groups

Abstract

The aims of this study were to test the ability of CE-SSCP in describing the variability of the drgestive contents flora of SPF (Specified Pathogenic Free) pigs from our experimental husbandry and to reach , by mrxing individual samples , the concept of a digestive flora 's profile characteristic of a batch of pigs. The faeces of s1x SPF sows were sampled and extracted DNA were m1xed to constitute more and more composite samples. In addition , the caecal contents of 12 SPF piglets, 1ssued from a srngle sow, were collected after slaughter at 28, 56 and 84 days postpartum and rndividually or after pooling tested. The DNA of each sample was extracted using the QIAamp DNA Stool Min1kit. The PCR for amplifying the rONA 16S V3 region was carried out on the individual DNAs and the mrxed DNAs corresponding to each date of slaughtering. Then the PCR products were analysed by CE-SSCP The reproducibility of the method has been tested first, and then the analysis of the profiles obtarned from the faeces of the swine was conducted. It described a withinindividual variability of about 40% when comparing SPF sows. When we mixed samples, variability between the profiles decreases with increasing number of faeces constituting the pool of DNA. Concernrng the p1glets, for each date of sampling , the pool of ccecal contents DNA defined a characteristic group profile. Moreover, this profile vaned with the age of the prglets These results confirmed the use of CE SSCP as a tool for the description of the of digestive flora balances and their evolut1on at the batch level. Thrs will be of particular interest in both animal health or food hygrene contexts associated wrth drgestive flora perturbatron .