The effects of being fed lauric acid on rumen characteristics were evaluated in a double 3 × 3 Latin square design using six Holstein steers with ruminal cannulas on a high grain diet. The steers were fed commercial concentrate (8.7 kg/day/steer) with one of three levels of lauric acid (0, 25 or 50 g/day/steer) and timothy hay (1.8 kg/day/steer). The feed intake and digestibility were determined. Ruminal fluid was collected at 3 h after feeding to determine chemical, physical and microbial parameters. An in vitro pure culture study was performed to determine the effects of lauric acid on Streptococcus bovis, a potent bloat- and acidosis-promoting rumen bacterium. There were no differences in feed intake and digestibility among the treatments. The proportion of butyrate and the viscosity of the rumen fluid tended to be lowered (P < 0.08 and P < 0.09, respectively) and the stable ingesta volume increase was significantly decreased (P < 0.01) by the lauric acid feed. The abundance of protozoa and bacteria did not differ among the treatments. In the in vitro study, the growth of S. bovis was inhibited by the lauric acid (100 nmol/L) but it showed an adaptive growth to lauric acid in long-term subculturing. The S. bovis that had adapted to lauric acid showed decreased viscosity and lactate production (P < 0.01) in culture with sucrose. These results indicate that supplemental lauric acid added to a high grain diet improves physical properties, possibly by altering the metabolic activity of S. bovis, and it may prevent the occurrence of feedlot bloat and acidosis in beef cattle.