Session Title: The Animal Microbiome
Presentation Date: Monday, August 18, 2008
Poster Board Number: 0360
DIVERSITY OF MICROBIAL FLORA IN PORCINE FECAL SAMPLES
K.-K. Cho1, M.A. Jeong1, C.H. Yun2, S.H. Kang3, B.U. Kim1
1Jinju National University, Department of Animal Resources Technology, Jinju, Korea, Republic of, 2Seoul National University, Department of Agricultural Biotechnology, Seoul, Korea, Republic of, 3Livestock Industry, CSIRO, Brisbane, Australia
Background and aims: Recent studies focusing porcine intestinal microbes directed to gain more insight on enteric diseases in relation to growth performance. However, culture-dependent microbiological methods have limitations in their practical use, for instance large scale screening of beneficial or harmful microorganisms. Therefore the present study aimed to develop the culture-independent assay based on analyzing 16s rRNA gene by high-throughput pyrosequencing technique.
Methods: We have corrected fecal samples from pigs at different ages, with/without diarrhea and with/without antibiotics and analyzed partial gene sequencing for 16s rRNA from microbes. Total DNA was extracted from fecal sample, bacterial 16s rRNA gene was amplified and cloned using prokaryote universal primer followed by Genome Sequencer-FLX pyrosequencing and taxonomic alignment.
Results: 24-week old healthy pigs fed no antibiotics contained 1,840 bacterial phylotypes and most of these were turned out to be Proteobacteria (41.1%), Firmicutes (22.8%), Actinobacteria (14.1%), Bacteroidetes (13.3%). Such pattern appeared to be similar regardless of antibiotic supplementation or diarrhea. Bacteroidetes were contained more than Firmicutes in terms of relative abundance of the sequences. It is to note that Proteobacteria were increased whereas Bacteroidetes and Firmicutes were reduced in healthy 7-week old pigs supplemented with antibiotics.
Conclusions: We have examined microbial 16s rRNA using pyrosequencing and taxonomic alignment on microbes in fecal samples from pigs at different ages, with/without antibiotics supplementation, and absence or presence of diarrhea. Major phyla of microbes in fecal samples were similar among different stages and treatment among pigs while there are differences on the abundance of the sequences.