Non-protein nitrogen supplementation on in vitro fermentation profile, methane production, and microbial nitrogen synthesis in a corn silage-based substrate.

Non-protein nitrogen (NPN) supplements improve animal performance in backgrounding diets. However, there is scarce information regarding the effect of different NPN sources and combinations on ruminal fermentation profile. The current study aimed to evaluate the effect of different NPN sources and their combinations on in vitro fermentation, microbial N synthesis, and methane (CH₄) production in a backgrounding diet. Incubations were conducted on three separate days for 24 h using corn silage and cotton gin byproduct (70% and 30% of DM, respectively) as substrate. Treatments were control (without NPN), urea, and five different proportions of urea-biuret and nitrate (100:0, 75:25, 50:50, 25:75, and 0:100). Each treatment, except control, was formulated to be isonitrogenous and equivalent to 1% urea inclusion. Ruminal fluid was collected from two ruminally cannulated Angus crossbred steers fed ad libitum corn silage and cotton gin byproduct plus 100 g of a urea-biuret-nitrate mixture. The concentration of volatile fatty acids (VFAs) and ammonia nitrogen (NH₃-N) were determined at 12 and 24 h of incubation. Final pH, in vitro dry and organic matter digestibility, total gas production, and concentration of CH₄ were determined at 24 h. The supplementation of NPN increased (P < 0.05) the concentration of NH₃-N at 12 and 24 h. Although NPN supplementation increased (P < 0.05) the concentration of total VFA and acetate at 12 h, treatments did not differ (P > 0.05) at 24 h. Supplementation of NPN increased (P < 0.05) the proportion of acetate at 12 and 24 h but tended to reduce (P = 0.054) the proportion of propionate only at 12 h. Digestibility and pH were not different (P > 0.05) among treatments. Increasing nitrates in the NPN supplement increased (P < 0.05) the proportion of acetate and reduced (P < 0.05) the proportion of butyrate at 12 and 24 h. The supplementation of NPN increased (P < 0.05) microbial N synthesis. Furthermore, increasing nitrate proportion in the NPN supplement increased (P < 0.05) the microbial N synthesis and efficiency of N use. Supplementation of NPN did not modify (P > 0.05) total gas or CH₄ production. However, increasing nitrate proportion in the NPN supplement linearly reduced (P < 0.05) CH₄ production. Supplementation of NPN increased NH₃-N concentration and microbial N while increasing the inclusion of nitrate decreased the production of CH₄ and increased the microbial N synthesis in a corn silage-based substrate under in vitro conditions.