Rumen fluid was taken from fistulated sheep and a cow receiving variou
s diets based on grass hay or grass cubes with and without cereal-base
d concentrates. Proteinases in the extracellular fluid, and extracted
from particulate material by Triton X-100, were visualized using SDS-P
AGE in which gelatin was co-polymerized with the gels. Each animal sam
pled had a different pattern of proteinase activity. No single protein
ase band predominated, although a few appeared in several samples, ind
icating that some microbial species were commonly involved in proteoly
sis but none was dominant. The banding patterns in samples taken from
the same animals two weeks apart were fairly similar, indicating some
stability within animals. Patterns obtained with extracellular fluid a
nd Triton X100 extracts of small and large particulate material from t
he same sample were similar for the most part, although the relative i
ntensity of the bands differed. The serine protease inhibitor, phenyl
methyl sulphonyl fluoride, had little influence on the banding pattern
. Concentrate in the diet appeared to increase inter-animal variation,
and sheep in adjacent pens and consuming the same grass hay:concentra
te diet had different banding patterns. Thus, no microbial proteolytic
enzyme was predominant in protein digestion in the rumen.