PSVIII-16 - Metagenomic Analyses of Microbial and Carbohydrate-active Enzymes in the Rumen of Holstein Steers Supplemented with Seeds of Pharbitis Nil

Abstract: Our previous research revealed the effects of dietary supplementation of Pharbitis nil seeds (PA) in improving the ruminal methane reduction and digestibility in Hanwoo steers. The objective of this study was to further investigate the effects of PA supplementation on rumen microbial taxonomy, Kyoto Encyclopedia of Genes and Genomes (KEGG) Orthology functional categories and the diversity of carbohydrate-active enzymes (CAZymes). Five rumen cannulated Holstein steers (744 ± 35 kg) were fed either basal diet (Control) or basal diet supplemented with PA at 5% dry matter intake (PA5) for two consecutive periods of 40 days in pairwise comparison. Rumen fluid was collected on the last day of each period after 4h of morning feeding. Analysis of rumen microbial metagenome revealed that contigs assigned to taxa Methanobrevibacter millerae decreased (p = 0.090) in PA5 group. Functional annotation of contigs using KEGG database revealed that genes involved in starch and sucrose metabolism were enriched (p = 0.058) in PA5 group compared with control. CAZyme analysis identified genes that encode glycoside hydrolase (GH) accounted for 63.18% and 65.66% of all CAZymes in control and PA5, respectively. The GH2 family (enriched in oligosaccharide-degrading enzymes) genes were greater (p = 0.090) in PA5 than control. Likewise, the abundance of GH26 and GH53 (endo-hemicellulases), GH5_4 (cellulases), and GH13_36 and GH13_42 (α-amylase) were increased (p < 0.1) in PA5 relatively compared with control. Most of the genes were mainly produced by unclassified contigs followed by Cupriavidus oxalaticus and Prevotella ruminicola 23. De novo assembly and binning recovered a total of 107 metagenome-assembled genomes taxonomically assigned to different bacterial phyla and one archaeal phyla. Of these genomes, 39 represented the uncultivated bacteria and 1 represented Lactobacillus genera. This study revealed that PA supplemented steers had increased GH activity, which encode enzymes that efficiently degrade plant cell wall polysaccharides in the rumen thereby increasing the fiber digestibility.