An in vitro assay was used to examine biotransformation of toxic Senecio jacobaea pyrrolizidine alkaloids (PA) in ovine, bovine, and caprine rumen contents. Pyrrolizidine alkaloids were analysed by high performance liquid chromatography, and the rates of the alkaloid biotransformation were determined. The microbiological "Most Probable Numbers" technique was also used, in combination with thin-layer chromatography, to estimate relative numbers of rumen PA-biotransforming bacteria in the same samples. Pyrrolizidine alkaloids were biotransformed at average rates of 2.9 micrograms/ml/h (bovine), 25.6 micrograms/ml/h (caprine), and 19.2 micrograms/ml/h (ovine). Estimates of numbers of PA-biotransforming bacteria were 1.1 x 10(7) bacteria/ml rumen contents (bovine), 2.4 x 10(7) bacteria/ml (caprine), and 3.0 x 10(7) bacteria/ml (ovine). This project is among the first to quantitate rates of PA biotransformation in rumen contents and to identify caprine and bovine, in addition to ovine, rumen PA-biotransforming activity, as well as to estimate the actual numbers of PA-biotransforming bacteria in rumen contents.