Multiresistant enterococci were tested for susceptibility to trimethoprim (TMP). Although most enterococci are inhibited by less than or equal to 1.0 microgram/ml, the MICs for 7 of 29 selected multiresistant isolates were greater than or equal to 8 micrograms/ml, including for two beta-lactamase positive (Bla+) strains, for which the MICs of TMP were greater than 1,000 micrograms/ml, and for another Bla+ strain, for which the MIC was 128 micrograms/ml. None of five isolates tested transferred TMP resistance and none of the resistant isolates hybridized to the dfrA gene of Staphylococcus aureus. Whether TMP resistance in enterococci is due to a mutation(s) or to acquisition of a new gene is not known. Acquisition of resistance to TMP is another example of the multiple antimicrobial resistance typically displayed by enterococci.