16S rRNA gene-based molecular methods to minitor Clostridium cluster Ⅳ community in the colon of piglets

Abstract:

Abstract: [Objective] To investigate the change of Clostridium cluster Ⅳ community in the colon of piglets from 7 to 35 days of age, and its correlation with butyrate concentration. [Methods] Three litters of neonatal piglets were used in this study. One piglet from each litter was sacrificed randomly at the age of 7, 14, 21（weaning day）, 24 and 35 days, digestive samples in the colon were collected. The concentration of volatile fatty acid(VFA) were determined by gas chromatography. 16S rRNA gene-based denaturing gradient gel electrophoresis (DGGE)and real-time PCR were used for qualitative and quantitative analysis of Clostridium cluster Ⅳ community. [Results] Similarity analysis of DGGE profile revealed that samples from piglets at the age of 7 days formed a coherent cluster with indices above 90%, no significant changes in Clostridium cluster Ⅳ community were found around weaning period. Real-time PCR analysis showed that 16S rRNA gene copies of total bacteria in the colon of piglets decreased significantly 3 days after weaning, this tendency was in accordance with the changes in concentration of total VFA and butyrate in colon, while there was no significant difference in copies of Clostridium cluster Ⅳ group. Sequencing analysis indicated that Clostridium cluster Ⅳ group in the colon of piglets were dominated by Faecalibacterium prausnitzii, Subdoligranulum variabile and other uncultured bacteria. [Conclusion] Changes in Clostridium cluster Ⅳ community in the colon of piglets were found from the age of 7 days to 14 days, while there was no significant difference during the weaning transition.