7). T. mobilensis hydrogenosomes
exhibit a flat peripheral vesicle ( Fig. 7a and c) whereas T. foetus presents a much more prominent and larger vesicle ( Fig. 7b and d). Because the T. mobilensis population was pleiomorphic, DNA analyses were performed to verify if any contamination by other tritrichomonads had occurred in T. mobilensis cultures. A molecular strategy previously described ( Kleina et al., 2004) was employed. For this purpose, the total DNA was extracted from two independent cultures and the rDNA ITS-1/5.8S/ITS-2 BMS-354825 molecular weight region was amplified. The PCR products were directly sequenced. As a result, both sequences obtained were identical to T. mobilensis isolate M776 (ATCC 50116) sequence retrieved from GenBank (U86612), thus, demonstrating that we were working with a T. mobilensis and that contamination did not take place. To compare the behavior of the different shapes of T. mobilensis, adherence assays using uncoated polystyrene microspheres were performed. Interestingly, quantitative analyses revealed that no differences were found in all parasites shapes analyzed ( Fig. 8). In addition, the binding capability of T. foetus was significantly higher than the binding capability of T.
mobilensis (P-value <0.01 by two-way ANOVA test) because approximately 40% (S.D. ± 3.42%) and 58% (S.D. ± 2.73) of the parasites from the cultured and fresh T. foetus isolates contained latex beads attached to their cell surface, respectively, whereas approximately
23% (S.D. ± 2.53%) of the parasites from both T. mobilensis isolates contained latex beads attached to their cell surface. Similar to T. foetus (data not buy Galunisertib shown), T. mobilensis presented binding capacity even during mitosis ( Fig. 8). To quantitatively assess the cytotoxicity of both species to Caco-2 cells, spectrophotometric analyses after MTT viability assays (Fig. 9) and crystal violet test (not shown) were performed. The MTT assay was carried out in the initial hours to follow the cytotoxic effects (Fig. 9). Both species were able to reduce the viability of Caco-2 Sodium butyrate cells. After 1 h of interaction, both strains from T mobilensis and T. foetus presented a similar cytotoxicity level. However, after 3 h, the cytotoxicity of both the cultured T. foetus (K strain) and T. mobilensis 4190 strain was higher than that of the fresh isolate of T. foetus (CC09-1) T. mobilensis USA:M776 strain ( Fig. 9). There are several studies on T. mobilensis concerning its pathogenicity, but only a few reports on the morphological aspects of this parasite. Therefore, it is important to have additional data based on ultrastructural studies as presented by this work. Both morphology and cytotoxicity assays of T. mobilensis were performed and compared with T. foetus in this study. To our knowledge, this is the first time that the morphological features of T. mobilensis and T. foetus have been compared. Culberson et al. (1986) showed that T.