Effects of bromoethansulfonic acid (2-BES) and monensin (M) on caecal fermentation pattern of fasted (F) and non-fasted (NF) rabbits (14 in total), were studied using in vitro batch incubations. In the first experiment, caecal contents were collected from 4 F (24 h) and 4 NF conventional rabbits (70–77 days old). They were incubated for 24 h at 39°C under 100% CO 2 or 50% CO 2 +50% H 2 atmosphere with 2-BES (1–20 mM) and monensin (50 mg/kg) and net production of volatile fatty acids (VFA) and methane (CH 4) was measured and hydrogen recoveries (2Hr) calculated. In fasted rabbits, 2-BES addition (1 mM) caused a slight decrease in CH 4 (14%) without changing the fermentation pattern whereas M caused a decrease in CH 4 (51%) accompanied by a decrease in total VFA production (29%), mainly butyrate (B) (37%) and acetate (A) (33%). In NF rabbits, 2-BES (20 mM) depressed CH 4 (93%) while M induced a surprising increase in CH 4(56%) coupled to a decrease in total VFA (16%), mainly B (34%). The consequent increase (31%) in the calculated hydrogen recoveries (2Hr) was hypothesized to be due to a decrease in acetogenic and reductive acetogenic bacteria. In the second experiment, hydrogen gas addition to caecal contents from 3 NF rabbits increased total VFA production (12%), mainly butyrate (32%) and monensin induced an increase in CH 4 production (33%) and changed the fermentation pattern towards lower total VFA, lower B and higher P. As microbial counts did not reveal a decrease in acetogenic and reductive acetogenic bacteria when monensin was added, it was hypothesized that the increase in CH 4 was due to a depression of their autotrophic activity, leaving more H 2 available for methanogenesis and reductive acetogens survived than heterotrophically.
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