Biological and biochemical studies for characterization of some Egyptian Trichomonas vaginalis isolates

Trichomoniasis has important medical, social and economical implications regarding its serious potentially associated complications, with the possibility of HIV acquisition and transmission. T. vaginalis is a very complex organism. Studying the variation in some biological and biochemical properties of the parasite can be used for characterization of the parasites. For the characterization of T. vaginalis infecting Egyptian female patients, the growth kinetics of 20 isolates, their pathogenicity, metronidazole susceptibility and electrophoretic protein patterns, were correlated with the recorded clinical manifestations associated with these isolates. Positive samples for T. vaginalis were cultured on modified Diamond`s medium. For growth pattern study, trophozoites were counted for each isolate every 24 hours for seven days. The pathogenicity assay was performed using intra-peritoneal inoculation in mice. The isolates susceptibility to different concentrations of metronidazole was recorded by determining the minimal lethal concentration [MLC] and trophozoites viability. The biochemical variability of the studied isolates was performed using 2-dimensional electrophoresis. A broad experimental variability was recorded among the 20 T. vaginalis isolates. There was a clear relationship between 3 isolates obtained from patients with severe vaginitis and the different parameters studied. These isolates had the highest number [20-25 organisms/HPF] in wet mount of vaginal discharge, and the shortest generation time [6:34-7:31 hours]; they were also highly pathogenic to mice. Only one isolate [no. 5] proved to be metronidazole resistant. The use of the first dimensional native-polyacrylamide gel electrophoresis [Native-PAGE] demonstrated the presence of some differences. The isolates were classified into two groups according to their proteins net charge. All samples in each group were considered as one isolate. However, when the 2-dimensional electrophoresis [SDS-PAGE] was applied, five different groups could be identified according to proteins molecular weights. There is a broad experimental variability among the studied Egyptian T. vaginalis isolates regarding growth kinetics, metronidazole drug susceptibility, degree of pathogenicity, as well as the electrophoretic protein patterns

Validity of igG avidity enzyme linked immunosorbent assay and polymerase chain reaction for the determinatipn of toxoplasma infections during pregnancy

Accurate diagnosis of maternal toxoplasmosis can enhance the success of medical treatment and prevent congenital transmission. The current diagnostic methods have significant limits and they poorly differentiate between acute and chronic infections. The present study was conducted to diagnose acquired Toxoplasma infection in pregnant women by detecting both the low avidity-IgG using enzyme linked immunosorbent assay [ELISA] and the specific DNA by polymerase chain reaction technique [PCR]. The study was performed on 104 pregnant women at different gestational ages. The serum sample from each case was primarily screened for IgG by ELISA, and then positive samples were examined for IgM and estimation of IgG avidity ELISA. The PCR technique was performed on blood samples from twenty selected cases, ten of whom were positive for both IgG and IgM antibodies and the other ten were positive for IgG only. Out of 104 pregnant females screened by IgG ELISA 48 cases were positive. Twenty seven of these cases were positive for IgG only and 21 had both IgG and IgM antibodies. Of the 27 IgG positive cases 24 showed high IgG avidity while the remaining three showed inconclusive avidity. Of the 21 cases positive for both IgG and IgM antibodies 16 patients showed low avidity while five cases only showed high avidity. PCR was done for ten cases positive for both IgG and IgM antibodies and ten positive for IgG only. It proved positive in only four often cases who were positive for both IgG and IgM and had low IgG avidity. This study highlights the value of IgG avidity ELISA as a single serum sample test indicating the presence of infection and differentiating between acute and chronic infections. It is recommended as a routine test for every pregnant woman. A positive PCR can act as an excellent indicator of recent infection, but its disadvantage lies in the fact that negative results cannot exclude acute infection. In addition it is an expensive technique. Thus IgG avidity ELISA can be considered as a more appropriate technique

newapproach in cultivation of leishmania major using human urine

Serum-free media have been devised for several Leishmania species, yet they tend to be complex and not particularly cheap. In the present work 5% healthy human urine was tried as a supplement to RPMI-1640 basal medium and was compared to 10% fetal calf serum. It greatly aided the growth of L.major invitro, and its maintenance for seven subcultures with five days interval. Infectivity of promastigotes was then tested by inoculation into mice. The lesion was similar in onset, duration and severity to the control group. However the urine-supplemented medium was unsuccessful for primary isolation of Leishmania from infected mice

effect of changes in the gastric PH value on experimental Trichinosis

The effect of increasing or decreasing the gastric PH of mice on the course of experimental infection with Trichinella spiralis was studied, by administration of 1% NaHCO[3] in PBS [PH 9.0] or 1% HC1 in PBS [PH 5.0] to mice half an hour prior to infection respectively. The results revealed that raising the gastric PH led to a significant increase in the adult worm count with an increase in their fecundity both in vivo and in vitro. On the other hand, the more acidic gastric PH induced prior to infection led to its amelioration. This was obvious by the significant reduction in the adult worm count and their inability to give birth to newborn larvae. Several factors may be incriminated, among which are decreased larval infectivity and affection of the maturation of the reproductive organs, mainly the uterus and the testis. Changes in their morphology were observed by both light and transmission electron microscopic studies, which could account for the impairment in their functions, namely embryogenesis and spermatogenesis