Multi-additive strategies with nitrate, fumarate, and probiotics to modulate ruminal methanogenesis and its effects on fermentation and microbiota in vitro

This study investigated the methane mitigation effects of three factors: nitrate (5 mM), fumarate (5 mM), and a multi-strain probiotic supplement (1.0*10^10 cells/mL), containing Lactiplantibacillus plantarum, Megasphaera elsdenii, Selenomonas ruminantium, and Acidipropionibacterium thoenii) in an in vitro rumen fermentation system. Rumen fluid was incubated with different treatment combinations, and microbial community profiles were analyzed by sequencing the V3-V4 region of the 16S rRNA gene using paired end Illumina sequencing.