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Several compounds were evaluated in vitro as alternative electron sinks to ruminal methanogenesis. They were incubated with ruminal fluid, buffer mixture, and finely ground alfalfa hay for 24 h, at 0, 6, 12, and 18 mM initial concentrations. The propionate enhancer oxaloacetic acid, the butyrate enhancer beta-hydroxybutyrate, and the butyrate unsaturated analog 3-butenoic acid were ineffective in decreasing methanogenesis. Nevertheless, beta-hydroxybutyrate increased apparent fermentation of the alfalfa hay substrate from 58.0 to 63.4%, and 3-butenoic acid seemed to increase it from 62.0 to 73.7%. Almost all of added oxaloacetic acid disappeared during the incubation, while only between 30.3 and 53.4% of beta-hydroxybutyrate disappeared. The butyrate enhancers acetoacetate and crotonic acid, and the butyrate unsaturated analog 2-butynoic acid, decreased methanogenesis by a maximum of 18,9 and 9%, respectively. Crotonic acid at 18 mM initial concentration seemed to increase the substrate apparent fermentation from 57.0 to 68.2%. Between 78.6 and 100% of acetoacetate disappeared during the incubation. The propionate unsaturated analog propynoic acid, and the unsaturated ester ethyl 2-butynoate, decreased methanogenesis by a maximum of 76 and 79%, respectively. Less than 5% of propynoic acid disappeared. The substrate apparent fermentation was decreased by propynoic acid from 62.0 to 57.4%, and seemed to have been decreased by ethyl 2-butynoate from 62.0 to 29.3%. More accurate measurements of the disappearance of some of the compounds studied are needed to better understand how they are metabolized and how they affect fermentation.