Effects of Isoacid Supplementation on In Vitro Rumen Fermentation, Nutrient Degradability and Bacterial Community...

This study investigated the effects of isoacid supplementation on in vitro rumen fermentation characteristics, nutrient degradability, and bacterial community diversity in yaks using corn silage-highland barley straw-based substrates. An in vitro fermentation experiment was conducted with a substrate consisting of 80% whole-plant corn silage and 20% highland barley straw. Treatments included a control (without isoacids) and four isoacid supplemental levels (0.1%, 0.2%, 0.3%, and 0.4% of substrate dry matter, DM), each with six replicates. A 72 h in vitro gas production experiment was performed to measure cumulative gas production, fermentation parameters, nutrient degradability, and bacterial community diversity. Cumulative gas production increased by 12.96% with 0.2% isoacid supplementation compared to the control (p < 0.05). The contents of microbial protein (MCP), acetate, propionate, and total volatile fatty acids (TVFA) exhibited quadratic responses to the increasing isoacid dosage (p < 0.05). Specifically, MCP content reached a maximum of 0.76 mg/mL with 0.2% isoacids, representing a 31.03% increase compared to the control (p < 0.05). TVFA content was highest (146.85 mmoL/L) at 0.2% isoacid supplementation, with a 16.40% increase compared to the control (p < 0.05). Acetate content increased by 17.99% (p < 0.05), while propionate tended to increase with 0.2% isoacid supplementation (p = 0.08). Supplementation with 0.2% and 0.4% isoacids did not alter the bacterial composition and diversity (p > 0.05). However, at the genus level, g_Ruminococcus, g__Elusimicrobium, g_norank_f_Atopobiaceae, g_norank_o_Coriobacteriales, and g_Romboutsia were identified as differential biomarkers showing significant responses to isoacid supplementation (p < 0.05). Mantel-test analysis revealed positive correlation between g_Ruminococcus abundance and NH₃-N content (r < 0.4, p < 0.05); g_Romboutsia abundance and acetate content (r < 0.40, p < 0.05); g_Defluviitaleaceae_UCG-011 abundance and both NH₃-N content and the pH of rumen fluid (r < 0.40, p < 0.05); g_norank_o_Coriobacteriales abundance and rumen pH (r < 0.40, p < 0.01). Supplementation with 0.2% isoacids to corn silage-barley straw substrates improved in vitro rumen fermentation characteristics in yaks, which was associated with altered abundances of key bacterial genera including g_Ruminococcus, g__Elusimicrobium, g_norank_f_Atopobiaceae, g_norank_o_Coriobacteriales.