The motility of viable Trichomonas vaginalis organisms is readily demonstrable in a clinical wet mount or cultured specimen. We attempted to determine whether migration is a dynamic process such that the organisms move to avoid exposure to toxic antimicrobial agents. With the use of axenic cultures of T. vaginalis that were radiolabeled and assayed for chemotaxis in plastic multiwelled plates with a membrane filter inserted to trap organisms, the response of clinical isolates to various antimicrobial agents was studied. Chemotaxis was readily demonstrable and dependent upon factors including time of incubation, media used, and viscosity of media. Nitroimidazoles (e.g., metronidazole) which readily inhibited the growth of these organisms also caQised significant chemorepulsion after minutes of exposure. The antifungal imidazoles ketoconazole and miiconazole inhibited growth neatly as readily and caused chemorepulsion, but to a lesser degree. The spermicide Nonoxynoi-9 also inhibited growth and caused significant chemorepulsion. The minitnal concentrations of many compounds which inhibited growth were very similar to those which caused significant chemorepulsion. Imidazole and antibiotics (e.g., penicillin) which did not inhibit growth did not induce any chemotactic effects. Chemotaxis of T. vaginalis is an active and dynamic process, and the organisms display chemorepulsion shortly after exposure to toxic antimicrobial agents, well before toxicity can be demonstrated.Numerous imidazole compounds have activity against Trichomonas vaginalis, and occasional reports of resistance to these compounds (e.g., the 5-nitroimidazole metronidazole) explain some cases of clinical failure (6, 10, 11).Because treatment failures seem to be far more common than the presence of resistant organisms (5,8,11), the motility of these organisms as a possible virulence factor was postulated by using the precedent of motility of Vibrio cholerae and the pathogenesis of cholera (4, 16).A recent report on the effect of hormones on T. vaginalis described a new assay system for the chemotaxis of this organism and noted again how motile they are (15). An attempt was made to better define the chemotaxis of these organisms and to study the effects of antimicrobial agents on this process.(This work was presented in part at the 88th Annual Meeting of the American Society for Microbiology, Miami Beach, Fla., 1988.) MATERIALS AND METHODS T. vaginalis. Isolates were cultivated from women with positive saline wet mounts examined for T. vaginalis by light microscopy. Sterile cotton swabs were used to inoculate secretions into borosilicate glass screw-cap tubes (8 by 100 mm) containing 6 ml of Dianmond DL8 defined medium, pH 7.2, supplemented with 8% human serum, 40 ,ug of gentamicin per ml, and 2.5 ,ug of amphotericin B (3, 7) per ml. Cultures were maintained in DL8 with 4% heat-inactivated equine serum at 36°C and were subcultured every 3 to 4 days (after reaching the stationary phase). Antibiotics were removed from the medium after several passages...