OxyraseTM for Optimal Growth of Trichomonas vaginalis Isolates |Biomedgrid

The inability of some fresh clinical isolates of Trichomonas vaginalis to grow and multiply when inoculated into the In-PouchTM trichomonas growth medium followed by death upon inoculation into the trypticase-yeast extract-maltose (TYM)-horse serum complex medium was investigated. Of fifty-one fresh clinical isolates derived from patients with trichomonosis, 11 (~20%) failed to grow to numbers after passage into the complex medium sufficient for cryopreservation (labeled Group 1). Isolates labeled Group 2 (~80%) thrived in both the In-PouchTM and complex media (Group 2). Experiments using the OxyraseTM scavenging system to remove oxygen from the medium revealed that Group 1 isolates grew to numbers equal to Group 2 isolate trichomonads. Group 1 isolates consisted of both dsRNA virus-negative and positive isolates, showing that absence or presence of virus was not responsible for the growth characteristics of the Group 1 isolate organisms. This further indicated that the Group 1 isolate parasites are unable to survive in a microaerophilic environment compared to Group 2 isolate trichomonads, suggesting that the patient vaginal environment was anaerobic. Analysis by one-dimensional sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of total proteins of T. vaginalis to compare Group 1 and Group 2 organisms showed no differences in protein patterns. Likewise, both Group 1 and Group 2 isolate parasites had similar total proteinase patterns by substrate gel analysis after electrophoresis. Comparative studies of the property of cytoadherence and amounts of adhesins also showed identical results for Group 1 organisms grown in OxyraseTM and Group 2 parasites in the presence or absence of OxyraseTM. These results are discussed in relation to the patient's vaginal anaerobic environment as critical for establishment of infection by Group 1 isolate parasites. This work now allows for investigators to perform studies of T. vaginalis isolates heretofore unable to be cultured and cryopreserved.