Assessment of reductive acetogenesis with indigenous ruminal bacterium populations and Acetitomaculum ruminis

The objective of this study was to evaluate the role of reductive acetogenesis as an alternative H2 disposal mechanism in the rumen. H2/CO2-supported acetogenic ruminal bacteria were enumerated by using a selective inhibitor of methanogenesis, 2-bromoethanesulfonic acid (BES). Acetogenic bacteria ranged in density from 2.5 x 10(5) cells/ml in beef cows fed a high-forage diet to 75 cells/ml in finishing steers fed a high-grain diet. Negligible endogenous acetogenic activity was demonstrated in incubations containing ruminal contents, NaH13CO3, and 100% H2 gas phase since [U-13C]acetate, as measured by mass spectroscopy, did not accumulate. Enhancement of acetogenesis was observed in these incubations when methanogenesis was inhibited by BES and/or by the addition of an axenic culture of the rumen acetogen Acetitomaculum ruminis 190A4 (10(7) CFU/ml). To assess the relative importance of population density and/or H2 concentration for reductive acetogenesis in ruminal contents, incubations as described above were performed under a 100% N2 gas phase. Both selective inhibition of methanogenesis and A. ruminis 190A4 fortification (>10(5) CFU/ml) were necessary for the detection of reductive acetogenesis under H2-limiting conditions. Under these conditions, H2 accumulated to 4, 800 ppm. In contrast, H2 accumulated to 400 ppm in incubations with active methanogenesis (without BES). These H2 concentrations correlated well with the pure culture H2 threshold concentrations determined for A. ruminis 190A4 (3,830 ppm) and the ruminal methanogen 10-16B (126 ppm). The data demonstrate that ruminal methanogenic bacteria limited reductive acetogenesis by lowering the H2 partial pressure below the level necessary for H2 utilization by A. ruminis 190A4.