Castro-Montoya, J. M., Makkar, H. P. S. and Becker, K. 2011. Chemical composition of rumen microbial fraction and fermentation parameters as affected by tannins and saponins using an in vitro rumen fermentation system. Can. J. Anim. Sci. 91: 433–448. Post-rumen chemical composition of the microbial fraction is one of the factors that determines the nutrients absorbed and available for maintenance and production of the animal. The hypothesis was that tannins and saponins alter chemical composition of rumen microbes and fermentation parameters in the rumen. Purified quebracho, mimosa, chestnut and sumach tannins; and quillaja and gypsophilla saponins were incubated with 380 mg of substrate (hay:concentrate 70:30 wt/wt) for 24 h in an in vitro gas production system at concentrations from 0.25 to 1.25 mg mL−1. Saponins increased N and reduced sugar contents of the liquid-associated microbes. The ratio of crude protein to purine bases significantly increased on adding sumach and chestnut tannins and decreased on the addition of quebracho and mimosa tannins. Quebracho, mimosa and chestnut tannins reduced total short-chain fatty acid production. The acetate:propionate ratio decreased for all additives. Results suggest that in vitro (a) depending on the source and the concentration, tannins would have an effect on the nitrogen and sugar contents of the liquid associated microbes, (b) saponins are likely to increase N and reduce sugar contents of rumen liquid associated microbes, and (c) estimation of microbial protein synthesis based on purine bases may lead to under- or over-estimations in the presence of tannins and saponins. In vivo studies are required to validate these results.