Abstract #LB6P

# LB6P
Synergistic associations of bacteria and archaea in DNA and cDNA components of rumen samples collected using stomach tube and cannula methods in dairy cows.
D. W. Pitta*1, C. F. A. Lage2,5, J. S. Bender1, N. Indugu1, M. L. Hennessy1, V. K. Shabtai1, B. Vecchiarelli1, A. Fernandez6, A. Spitzer7, S. E. Raisanen2, A. Melgar2, K. Nedelkov3, X. Chen4, J. Oh2, A. N. Hristov2, 1University of Pennsylvania, Kennett Square, PA, USA, 2The Pennsylvania State University, University Park, PA, USA, 3Universidade Federal de Minas Gerais, Belo Horizonte, Brazil, 4College of Pastoral Agriculture Science and Technology, Lanzhou University, China, 5Faculty of Veterinary Medicine, Trakia University, Stara Zagora, Bulgaria, 6Eberly College of Arts and Sciences, West Virginia University, Morgantown, WV, USA, 7School of Veterinary Medicine, Cornell University, Ithaca, NY, USA.

The rumen microbial ecosystem is comprised of bacteria, protozoa, fungi and archaea that work synergistically to facilitate feed digestion. However, the synergistic interactions between different microbes is seldom investigated. Here we investigated rumen bacteria and archaea associations of 6-cannulated Holstein cows that were adapted to a basal total mixed ration. Rumen samples were collected via stomach tube (ST) and rumen cannula (RC) at 0, 2, 4, 6, 8 and 12 h post feeding and filtered to collect solid rumen contents. These samples were extracted for genomic DNA (total) and RNA (cDNA; metabolically active), PCR amplified, sequenced and analyzed using QIIME pipeline for DNA and cDNA-based bacteria and archaea diversity. Based on PERMANOVA analysis for bacteria and archaea communities differed (P ≤ 0.05) between DNA and cDNA but showed no differences (P ≥ 0.05) between ST and RC. At the individual taxa level, ANCOM analysis showed that lineages of Actinobacteria and Bacteroidetes were lower (P ≤ 0.05) and those of Proteobacteria and Fibrobacter bacterial phyla were higher in the cDNA compared with DNA bacteria across ST and RC samples. Particularly, the abundance of Ruminococcus and Succinivibrionaceae were more than doubled in the cDNA compared with DNA bacteria. Similarly, for archaea, Methanobrevibacter was lower (P ≤ 0.05) and VadinCA11 (P ≤ 0.05) was higher in cDNA compared with DNA archaea in ST and RC samples. These results indicate that the cDNA approach is more discriminatory than the DNA approach for microbial diversity analysis in ST and RC methods. Correlation analysis of cDNA bacteria with molar proportions of volatile fatty acids in both ST and RC revealed strong positive correlations (P ≤ 0.05) between Ruminococcus and acetate, Succinivibrionaceae, Prevotella and Bulledia and propionate and Clostridium and butyrate. Co-occurrence analysis revealed that co-associations existed between Ruminococcus and Methanobrevibacter, and Succinivibrionaceae and Methanosphaera. This is the first study demonstrating the synergistic associations of bacteria-archaea cohorts in the rumen that are individual specific but differed between individual cows.

Key Words: bacteria-archaea cohorts, proxy, ANCOM