EVALUATION OF THE EFFECT OF IRON AND POLYAMINES ON TRICHOMONAS VAGINALIS VIRULENCE

Abstract

The aim of the present study is toinvestigate the in vitro effect of iron and spermine on the growth kinetics, cytoadherence and cytotoxicity of T.vaginalisand to examine the in vitro effect of iron on MLC of metronidazole on T. vaginalis isolates. In order to achieve this aim, vaginal washouts and urine samples were collected from 100 female patients attending the Early Cancer Detection Unit, and the out-patients clinics, Gynecology and Obstetrics Hospital, Faculty of Medicine, Ain Shams University.They were examined for the presence of T. vaginalistrophozoites by wet mount microscopic examination and in vitro culture on TYM media. Two differentT. vaginalis isolates were maintained to conduct the study;maintenance was done by subculturing of T. vaginalis every 24-48 hours in new culture medium. Growth kinetics wasassessed by counting parasites grown on culture media with different conditions every 24 hours for 7 days. Parasites were grown on iron-rich culture medium supplemented with 250 µM FAS, iron-depleted culture medium supplemented with 100 µM 2,2-DP, TYM medium supplemented with 20mM DAB for polyamine metabolism inhibition and polyamine metabolism restoration was done by harvesting DAB-treated parasites after 18-hrs growth, washed with sterile PBS and transferred to TYM-serum medium supplemented with 50 µM spermine at 37 ºC. The cytotoxicity and cytoadherencelevels of trichomonads, grown on culture media with different conditions, over Vero cell were assessed. Aerobic and anaerobic metronidazole susceptibility assays were done to evaluate the effect of iron on MLC of metronidazole on T. vaginalis isolates.Susceptibility assays were done on T. vaginalis grown on normal and iron-rich media.