SUMMARY: The optimal conditions for accurate and reproducible analysis of cellulose hydrolysis by micro-organisms, particularly from the m e n , were determined. Rumen micro-organisms were found to require approximately 50 mg. substrate (cellulose powder) in a p0 hr. incubation at 3 7 ' with 1 ml. of m e n liquor, and a total volume of 8 ml., in static vessels. All types of cellulose or simpler substituted derivatives, from soluble carboxymethylcellulose through insoluble cellulose powder to cotton fibres, are nearly quantitatively hydrolysed by mixed rumen micro-organisms. The effect of the inorganic composition of the medium, of antibacterial agents (thymol, fluoride, toluene) and of concentration procedures on the cellulolytic activity of rumen micro-organisms was examined. Rumen protozoa appear to play a minor role in cellulose degradation; their effective activity is due mainly to closely associated, possibly ingested, bacteria. The validity of the results obtained by various methods of ' cellulase ' assay is discussed.