Trichomoniasis is a sexually transmitted disease of cattle and a large bowel diarrheal disease of cats caused by Tritrichomonas foetus. Recently, other species of trichomonads have been identified from the prepuce of virgin bulls. It is not clear whether these non-T. foetus isolates are common (nor) or is it clear whether they are also present on the prepuce of breeding bulls. To answer these questions, we first developed an immunofluorescent assay (IFA) with T. foetus-specific monoclonal antibodies for comparison with a T. foetus-specific PCR assay. Results showed that all PCR positive isolates were also IFA positive, whether the isolates were from cats or cattle and PCR negative isolates were IFA negative. Bovine non-T. foetus (non-Tf) trichomonad isolates were detected by both assays in 14 virgin bulls, 10 breeding bulls, 21 bulls of undetermined breeding status (presumably breeding bulls) and 2 cows. These isolates from virgin bulls were mostly Tetratrichomonas spp. whereas the non-Tf isolates from most breeding bulls and the two cows were Pentatrichomonas hominis. All T. foetus isolates were from breeding bulls or bulls of undetermined breeding status. This IFA test which discriminates between T. foetus and non-Tf may be useful as a diagnostic assay, since no effective legal treatment is available, bulls positive for T. foetus are culled. With increasing reports of T. foetus large bowel infection in cats, these monoclonal antibodies may also be useful for diagnosis of feline infection. Since two isolates of non-Tf trichomonads were obtained vaginas of breeding cows, it may be that these parasites are sexually transmitted like pathogenic T. foetus.