Modulatory effect of iron chelators on adenosine deaminase activity and gene expression in Trichomonas vaginalis

Trichomonas vaginalis is a flagellate protozoan that parasitises the urogenital human tract and causes trichomoniasis. During the infection, the acquisition of nutrients, such as iron and purine and pyrimidine nucleosides, is essential for the survival of the parasite. The enzymes for purinergic signalling, including adenosine deaminase (ADA), which degrades adenosine to inosine, have been characterised in T. vaginalis. In the evaluation of the ADA profile in different T. vaginalisisolates treated with different iron sources or with limited iron availability, a decrease in activity and an increase in ADA gene expression after iron limitation by 2,2-bipyridyl and ferrozine chelators were observed. This supported the hypothesis that iron can modulate the activity of the enzymes involved in purinergic signalling. Under bovine serum limitation conditions, no significant differences were observed. The results obtained in this study allow for the assessment of important aspects of ADA and contribute to a better understanding of the purinergic system in T. vaginalis and the role of iron in establishing infection and parasite survival.

Detección de Chlamydia trachomatis, esporos micóticos y Trichomonas vaginalis en mujeres en edad fértil que acuden a los Hospitales San Pablo y Regional de San Lorenzo / Chlamydia trachomatis, fungal spores and Trichomonas vaginalis detection of childbearing age women who attend to San Pablo Hospital and Regional of San Lorenzo

Introducción: Las infecciones vulvo-cérvico-vaginales constituyen la causa más común por la que las mujeres en edad fértil acuden al Ginecólogo. Los agentes etiológicos involucrados incluyen a Chlamydia trachomatis, Trichomonas vaginalis, Cándida albicans, entre otros. La sospecha de vulvo-cervico-vaginitis debe orientar a la realización de estudios de secreción vaginal y muestreo endocervical a fin de detectar el agente causal, establecer un tratamiento específico y disminuir la prevalencia de los mismos así como las complicaciones. Objetivos: Determinar la prevalencia de esporos micóticos, Trichomonas vaginalis y Chlamydia trachomatis en mujeres en edad fértil. Materiales y Métodos: Estudio observacional, descriptivo, de corte transversal y prospectivo, con cuestionario realizado a mujeres en edad fértil que acudieron a los hospitales Materno Infantil San Pablo y Regional de San Lorenzo entre los meses de diciembre de 2010 a febrero de 2011. Resultados: De las 148 pacientes estudiadas, se observó una prevalencia de 27,7% de esporos micóticos, 2% de Trichomonas vaginalis y 4,1% de Chlamydia trachomatis. De las pacientes estudiadas, la mayoría tenían entre 25 a 29 años (26%). Conclusión:La prevalencia de esporos micóticos fue 27,7%, Trichomonas vaginalis 2% y Chlamydia trachomatis 4,1%.

Introduction: the vulvo cervicovaginal infections are the most common reason that women of childbearing age go to the gynecologist. The ethiological agents involved are Chlamydia trachomatis, Trichomonas vaginalis, Candida albicans, among others. The suspicion of cervico-vulvo-vaginitis should guide the studies of vaginal discharge and endocervical sampling to detect the causative agent, to stablish specific treatment and reduce the prevalence of these infections and the complications. Objective: to determine the prevalence of fungal spores, Trichomonas vaginalis and Chlamydia trachomatis in women of childbearing age. Materials and Methods: observational, descriptive, cross-sectional and prospective study performed in women of childbearing age who attended the San Pablo Hospital and Regional of San Lorenzo between the months of December 2010 to February 2011. Results: 148 patients were studied, there was a 27.7% prevalence of fungal spores, 2% Trichomonas vaginalis and 4.1% Chlamydia trachomatis. Of the patients studied, most were between 25 to 29 years (26%). Conclusion: the prevalence of fungal spores was 27.7%, Trichomonas vaginalis 2% and Chlamydia trachomatis 4.1%.

Prevalence and Comparison of Diagnostic Methods for Trichomonas vaginalis Infection in Pregnant Women in Argentina

The objectives of this study were to conduct a prevalence survey of trichomoniasis in pregnant women and to evaluate the utility of different methods for its diagnosis. A total of 597 vaginal exudates from pregnant women who were examined at the Hospital de Clinicas in Buenos Aires, Argentina from 1 August 2005 to 31 January 2007, were prospectively and consecutively evaluated. The investigation of Trichomonas vaginalis was made by different microscopic examinations, and culture on liquid medium. The sensitivity and specificity of the microscopic examinations were assessed considering culture on liquid medium as the "gold standard". The prevalence of T. vaginalis obtained by culture on liquid medium was 4.0% (24/597). The prevalence of T. vaginalis obtained by direct wet smear, prolonged May-Grunwald Giemsa staining, and sodium acetate-formalin (SAF)/methylene blue staining-fixing technique was 1.8%, 2.3% and 2.5%, respectively. The sensitivity of the direct wet smear was 45.8%, that of the prolonged May-Grunwald Giemsa staining was 58.3%, and that of the SAF/methylene blue method was 62.5%. Considering the 3 microscopic examinations altogether, the sensitivity rose to 66.7% and the specificity was 100% for all of them. This is the first time that the prevalence data of T. vaginalis by culture in pregnant women are published in Argentina. Due to the low sensitivity obtained by microscopy in asymptomatic pregnant women, the use of the liquid medium is recommended during pregnancy, in order to provide an early diagnosis and treatment.

Evaluación de 4 medios de cultivo para aislamiento primario de trichomonas vaginalis / Evaluation of four media the primary isolation of trichomonas vaginalis

The incidence sexually transmitted diseases (STD) is a global public health problem, highly complicated by infections produced by the human immunodeficiency virus. Parasitic etiological agents of STD in Chile are limited to trichomonas vaginalis, sarcoptes scabiei and phthirus pubis. Only T. vaginalis has a trophozoite stage which can be diagnosed with a high degree of success in the laboratory. The present study reports on four culture methods which facilitate the primary isolation of this protozoan from clinical samples. The best results for primary isolation of T. vaginalis were obtained using modified diamond media. It was demonstrated that Kupferberg-Agar medium was the best for growth of T. vaginalis without re-inoculation. The optimal pH for the growth of T. vaginalis in the cultures was 6.5, and 10 percent horse serum gave the best yields in development and viability of T. vaginalis over time.

La incidencia de las enfermedades de transmisión sexual (ETS) se mantiene como problema de salud pública a nivel mundial, agravado por las infecciones por los virus de la inmunodeficiencia humana. Los agentes etiológicos parasitarios de ETS que existen en Chile se limitan a Trichomonas vaginalis, Sarcoptes scabiei y Phthirus pubis. T. vaginalis sólo posee estadio de trofozoito por lo que el diagnóstico de laboratorio es de alta exigencia. En este trabajo se evaluaron 4 medios de cultivo para facilitar el aislamiento primario de este protozoo a partir de muestras clínicas. Con el medio Diamond Modificado se obtuvo los mejores resultados para el aislamiento primario de T. vaginalis. Se comprobó que el medio Kupferberg-Agar fue el más adecuado para el crecimiento sin repique de T. vaginalis. Se determinó que el pH del medio de cultivo para el crecimiento óptimo de T. vaginalis, fue dr 6,5, y suero quino al 10 por ciento fue la concentración de mejor rendimiento para desarrollo y viabilidad en el tiempo de T. vaginalis.

Hydrogenosomal activity of Trichomonas vaginalis cultivated under different iron conditions

To evaluate whether iron concentration in TYM medium influence on hydrogenosomal enzyme gene expression and hydrogenosomal membrane potential of Trichomonas vaginalis, trophozoites were cultivated in irondepleted, normal and iron-supplemented TYM media. The mRNA of hydrogenosomal enzymes, such as pyruvate ferredoxin oxidoreductase (PFOR), hydrogenase, ferredoxin and malic enzyme, was increased with iron concentrations in T. vaginalis culture media, measured by RT-PCR. Hydrogenosomal membrane potentials measured with DiOC6 also showed similar tendency, e.g. T. vaginalis cultivated in iron-depleted and iron-supplemented media for 3 days showed a significantly reduced and enhanced hydrogenosomal membrane potential compared with that of normal TYM media, respectively. Therefore, it is suggested that iron may regulate hydrogenosomal activity through hydrogenosomal enzyme expression and hydrogenosomal membrane potential.

Avaliaçäo do emprego de soro humano no meio de Diamond para isolamento e manutençäo de cepas de Trichomonas vaginalis / Evaluation of the use of human serum in Diamond's Medium for the isolation and maintenance of Trichomonas vaginalis

O meio de Diamond vem sendo empregado para o cultivo do Trichomonas vaginalis, Trichomonas tenax e Pentatrichomonas hominis, em sua composiçäo utiliza-se soro de cavalo como fonte de enriquecimento. Neste trabalho, este soro foi substituído pelo soro humano, por facilidade na sua obtençäo em laboratórios de análises clínicas. Os meios de cultura, assim preparados, foram distribuídos em tubos com tampa de rosca, em alíquotas de 8 mL, e os repiques foram feitos a cada 5 dias. As cepas de Trichomonas vaginalis foram inoculadas concomitantemente em meios contendo soro de cavalo e soro humano; resultados satisfatórios foram obtidos no cultivo e manutençäo das cepas, e o número de parasitas dobrou no mesmo período quando foi usado soro humano. Esses dados mostram que o isolamento e a manutençäo desses flagelados podem ser realizados de maneira simples e rotineira nos laboratórios, possibilitando uma melhoria no diagnóstico dessa parasitose e facilidade na manutençäo de cepas para outros estudos.

Comparable growth of a Trichomonas vaginalis strain in PEHPS and TYI-S-33 media

PEHPS medium, developed for zxenic cultivation of Entamoeba histolytica and E. invadens, was also capable of supporting the growth of a Trichomonas vaginalis strain, with an inoculum of 1 to 100 trichomonads/ml. The lorithmic growth phase in PEHPS or in TYI-S-33 medium lasted 72 h; yield (3.33 ñ 0.56 x 10 a the 6 trichomonads/ml), duplication time (4.27 h), number of duplications (16.85), or increase ratio (33,328) in PEHPS medium showed no significant differences with those obtained in TYI-S33 under similar culture conditions. Accordingly, PEHPS medium might be used for the axenic cultivation of T. vaginalis

Crescimento homogêneo de trichomonas vaginalis em meio semi-sólido / Homogeneous growth of trichomonas vaginalis in a semi-solid culture medium

Apesar da Tricomoníase ser, geral, a doença sexualmente transmitida mais frequentemente diagnosticada, existem poucas informaçöes relacionadas a padronizaçäo de cultivo do agente etiológico em meio semi-sólido. No presente trabalho, 10 estirpes de Trichomonas vaginalis foram semeadas em meio semi-sólido de Diamaond modificado, estudando-se as concentraçöes de ágar, o inóculo e o tempo de incubaçäo que permitiam crescimento do parasita em todo o meio de cultura. Foi observado que inóculo de 10**5 ou 10**6 Trichomonas vaginalis por mililitro, concentraçäo de 0,6% de ágar e tempo de incubaçäo de 72 horas foram as condiçöes mais adequadas, para o crescimento homogêneo do referido agente, mediante a aplicaçäo da técnica do ágar-fundido

Meio de transporte para Trichomonas vaginalis / Transport mean to Trichomonas vaginalis

O Trichomonas vaginalis é detectado rotineiramente logo após a colheita do material biológico. Como esse procedimento torna-se inviável em algumas situaçöes, faz-se necessário o estudo de um meio de transporte para sua demonstraçäo. Para isso, a vitalidade do parasita foi estudada em soluçäo salina comum, soluçäo salina glicosada e tamponada a pH 6.0, 7.0 e 8.0 e meio de cultura às temperaturas de 25.27-C e 37-C. Os resultados obtidos demonstraram se a soluçäo salina glicosada tamponada com fosfato de grande valor como meio de transporte para o T. vaginalis em temperaturas de 25.27-C