We previously reported that Modified Field Stain (MF) can be used as a rapid stain for diagnosis. In the present study we extend the observation to include the stain as an alternative method to assess viability of the cells.
Trichomonas vaginalis, a flagellated protozoan parasite, is commonly found in the genitourinary tract of humans. Its mode of reproduction has always been reported to be binary fission. The high parasite numbers seen in a relatively short period in in vitro cultures led us to believe that there must be other modes of reproduction. The present study for the first time provides transformational evidence at the ultrastructural level seen in tropohozoites of T. vaginalis undergoing a multiple asexual mode of reproduction. The findings show that the single cell with a nucleus is capable of dividing to as many as eight nuclei within the cytoplasmic body. Before the commencement of division, the nucleus remained round or ovoid in shape with condensed chromatin masses and only a few endoplasmic reticula surrounding the nucleus. During the division, the nucleus started to elongate and become irregular in shape with visible chromatin masses condensing with the accumulation of numerous endoplasmic reticula. Nuclear division gave rise to as many as eight nuclei within a cell, which could be seen to be connected by numerous endoplasmic reticula. In addition, a high number of hydrogenosomes and vacuoles can be seen in multinucleated T. vaginalis compared with single nucleated T. vaginalis. This study confirms that multiple modes of nuclear division do exist in T. vaginalis and are a precursor to progeny formation.
The protozoan Trichomonas vaginalis a sexually transmitted protozoan parasite causes vaginitis, urethritis and cervicitis in humans. The present study highlights phenotypic 'variant' forms of trophozoites isolated from patients suffering from cervical neoplasia condition. The growth curve of 10 isolates i.e., four non-cervical neoplasia (NCN) isolates (NCN1-NCN4) and six cervical neoplasia (CN) isolates (CN1-CN6) showed two distinct and different in vitro growth profiles. The parasite count and growth rates were significantly higher in trophozoites from CN isolates in cultures of day 2 up to day 8 (p<0.05, Mann-Whitney test). The average generation time was 1.84±0.40 and 3.38±0.55h for NCN and CN isolates respectively. The nucleus of trophozoites in CN isolates using acridine orange and DAPI showed more intense staining revealing higher nuclear content. The FITC-labeled Concanavalin A stained stronger green fluorescence with surface of trophozoites in CN isolates showing more rough and creased surface with numerous deep micropores. Transmission electron microscopy studies revealed that there was higher numbers of vacuoles and hydrogenosomes in these forms. The study mounted staining techniques, growth profiles, morphology, morphometry studies using scanning and transmission electron microscopy and confirms that the trophozoites from cervical neoplasia proliferates at a higher rate, shows higher FITC-labeled Concanavalin A binding with rough and creased surface implying that these are virulent forms which can aggravate or exacerbate cervical neoplasia conditions. The large numbers of hyrogenosomes and vacuoles implies that these forms are active and implicates a possible role in such conditions.
Trichomonas vaginalis, a flagellated protozoan parasite causes a variety of adverse health consequences in both men and women. The parasite exists in the trophozoite and the pseudocystic stage. The study reports for the first time that pseudocyst forms of T. vaginalis isolated from cervical neoplasia (CN) patients demonstrated distinct, different and significant in vitro growth profiles when grown in vitro cultures from day 1 up to day 5 (p<0.05, Mann-Whitney test) when compared with the same life cycle stages isolated from non-cervical neoplasia but symptomatic patients (NCN). Pseudocysts from CN and NCN isolates remained viable in distilled water until 3 h 10 min and 2 h 10 min, respectively. The nucleus of pseudocysts in CN isolates using acridine orange and DAPI showed more intense staining revealing higher nuclear content. The FITC-labeled Concanavalin A stained stronger green fluorescence with surface of pseudocysts in CN isolates showing more rough and creased surface with higher numbers of deep micropores with larger numbers of chromatin masses, vacuoles, and hydrogenosomes. The study confirms that pseudocystic stage from CN, despite the uniformity in appearance of being rounded and showing no motility without a true cyst wall under light microscopy, demonstrated different biochemical, surface, and ultrastructural properties. The study provides evidence that phenotypic variant forms of pseudocysts does exist and possibly does play a role in exacerbating cervical cancer.
Wet mount microscopy is the most commonly used diagnostic method for trichomoniasis in clinical diagnostic services all over the world including Sri Lanka due to its availability, simplicity and is relatively inexpensive. However, Trichomonas culture and PCR are the gold standard tests. Unfortunately, neither the culture nor PCR is available for the diagnosis of trichomoniasis in Sri Lanka. Thus, it is important to validate the wet mount microscopy as it is the only available diagnostic test and has not been validated to date in Sri Lanka. The objective was to evaluate the validity and reliability of wet mount microscopy against gold standard Trichomonas culture among clinic based population of reproductive age group women in Western province, Sri Lanka. Women attending hospital and institutional based clinics were enrolled. They were interviewed and high vaginal swabs were taken for laboratory diagnosis by culture and wet mount microscopy. There were 601 participants in the age group of 15-45 years. Wet mount microscopy showed 68% sensitivity, 100% specificity, 100% positive (PPV) and 98% negative predictive values (NPV) (P=0.001, kappa=0.803) respectively against the gold standard culture. The area under the ROC curve was 0.840. Sensitivity of wet mount microscopy is low. However it has high validity and reliability as a specific diagnostic test for trichomoniasis. If it is to be used among women of reproductive age group in Western province, Sri Lanka, a culture method could be adopted as a second test to confirm the negative wet mount for symptomatic patients.
Kajian ini dilakukan untuk menentukan prevalens jangkitan Trichomonas vaginalis, di kalangan wanita yang berumur 20 hingga 59 tahun di Hospital Umum Sarawak melalui pemeriksaan ke atas lumuran Pap lazim. Prevalens jangkitan ini dikaji di kalangan lima kumpulan etnik utama iaitu kaum Iban, Cina, Melayu, Bidayuh dan Orang Ulu. Penyaringan dijalankan pada lumuran Pap lazim ke atas 300 sampel slaid yang telah diproses dan dicelup menggunakan pencelup Papanicolaou. Berdasarkan maklumat pada borang permohonan penyaringan lumuran Pap, tanda dan gejala jangkitan diambil kira sebagai petunjuk penting semasa penyaringan kerana melalui tanda dan gejala jangkitan, lumuran Pap mempunyai hubungan yang rapat kepada ciri-ciri gambaran sitomorfologi jangkitan yang berlaku ke atas sel epitelial sekiranya ianya mengalami jangkitan. Kategori umur untuk setiap golongan etnik juga dijadikan sebagai salah satu faktor pengukur bagi menilai tahap kekerapan jangkitan. Hasil kajian menunjukkan trikomoniasis mencatatkan jangkitan sebanyak 7.7% kes. Jangkitan berlaku pada kesemua kumpulan etnik dan lebih kerap ditemui pada golongan umur 20 hingga 29 tahun. Kesemua kes turut menunjukkan tanda dan gejala jangkitan. Hasil kajian ini menunjukkan kepentingan menjalani ujian lumuran Pap sebagai ujian mengesan jangkitan Trikomonas vaginalis selain saringan awal pengesanan kanser serviks.
In a prospective, observational study, 380 women attending a family planning (LPPKN) clinic and Sexually Transmitted Diseases (STD) clinic in Kuala Lumpur between January 2008 and November 2008 were enrolled. Patient information was obtained from the medical records and patients voluntarily completed questionnaires. Three vaginal swabs from the posterior fornix were taken from each patient examined. Different staining methods and cultivation in Diamonds medium were performed on the collected samples. Study subjects recruited in this survey were mostly young, with a geometric mean ages of 37.31 years (LPPKN clinic) and 32.06 years (STD clinic). Malay, Chinese, Indians and others ethnic groups accounted for 91, 4, 2 and 3%, respectively, at the LPPKN clinic. At the STD clinic, almost all subjects were Chinese. The prevalence rate of trichomoniasis was 0.36% at the LPPKN clinic and 0.0% at the STD clinic. Our findings suggested no association between contraceptive methods used and trichomoniasis infection at the LPPKN clinic. At the STD clinic, it was found a high risk for STD acquisition was not associated with trichomoniasis positivity. The data demonstrated the prevalence of trichomoniasis was very low at LPPKN clinic and absent at STD clinic. The high prevalence rates of other STD among sex workers found in this survey suggest the need for more active interventions focused on this group.
Trichomonas vaginalis, a flagellate protozoan parasite commonly found in the human genitourinary tract, is transmitted primarily by sexual intercourse. Diagnosis is usually by in vitro culture method and staining with Giemsa stain. There are laboratories that use Gram stain as well. We compared the use of modified Field's (MF), Giemsa, and Gram stains on 2 axenic and xenic isolates of T. vaginalis, respectively. Three smears from every sediment of spun cultures of all 4 isolates were stained, respectively, with each of the stains. We showed that MF staining, apart from being a rapid stain (20 s), confers sharper staining contrast, which differentiates the nucleus and the cytoplasm of the organism when compared to Giemsa and Gram staining especially on parasites from spiked urine samples. The alternative staining procedure offers in a diagnostic setting a rapid stain that can easily visualize the parasite with sharp contrasting characteristics between organelles especially the nucleus and cytoplasm. Vacuoles are more clearly visible in parasites stained with MF than when stained with Giemsa.
There is a demand for patients to self-diagnose their sexually transmitted infections (self- testing), particularly during the coronavirus pandemic to prevent infection spread. We enrolled a cohort of Saudi women in a single-visit prospective study, which was the first of its kind performed in the country. Our aim was to evaluate the OSOM® Trichomonas (OSOM) test, a single-use, point-of-care rapid test, for its efficacy and accessibility as a self-test for Trichomonas vaginalis (Trichomonas) infection. At a public hospital's gynecology clinic, women received sufficient training on specimen collection and OSOM self-testing. The women's infection status was re-evaluated using direct wet mount microscopy and clinician performed OSOM using additional swabs. Specimens with discordant results were sorted using an in- house polymerase chain reaction (PCR). 174 women aged 18 to 35 were registered and self- tested at the clinic under the supervision of a gynecologist between June and December 2020, with 84.4 percent (147/174) having a valid result on the first or repeat OSOM. Infection was detected in 12.2% (18/147) of participants, with two-thirds of them symptomatic. Young age, low education, the existence of vaginitis symptoms, and unemployment were identified as key risk factors for infection in the study population, with statistically significant differences seen among women only in terms of education level and employment status (p<0.001). The OSOM self-test performed well (83.3% sensitivity and 98.4% specificity), outperforming the wet mount microscopy (72.2% sensitivity and 100% specificity) and comparable to the clinicians' OSOM (88.8% sensitivity and 100% specificity). The patients' and clinicians' OSOM tests were strongly correlated, with a kappa of 0.89 and a 97.9% agreement. Self-collection of vaginal swabs was accepted and preferred by most women (94%) over the clinician-collection. Overall, our study's findings may have important consequences for the implementation of Trichomonas screening based on OSOM self-testing approach in the study's population.
Microporous polymeric matrices prepared from poly(ɛ-caprolactone) [PCL] were evaluated for controlled vaginal delivery of the antiprotozoal agent (tinidazole) in the treatment of the sexually transmitted infection, trichomoniasis. The matrices were produced by rapidly cooling co-solutions of PCL and tinidazole in acetone to -80 °C to induce crystallisation and hardening of the polymer. Tinidazole incorporation in the matrices increased from 1.4 to 3.9% (w/w), when the drug concentration in the starting PCL solution was raised from 10 to 20% (w/w), giving rise to drug loading efficiencies up to 20%. Rapid 'burst release' of 30% of the tinidazole content was recorded over 24 h when the PCL matrices were immersed in simulated vaginal fluid. Gradual drug release occurred over the next 6 days resulting in delivery of around 50% of the tinidazole load by day 7 with the released drug retaining antiprotozoal activity at levels almost 50% that of the 'non-formulated' drug in solution form. Basic modelling predicted that the concentration of tinidazole released into vaginal fluid in vivo from a PCL matrix in the form of an intravaginal ring would exceed the minimum inhibitory concentration against Trichomonas vaginalis. These findings recommend further investigation of PCL matrices as intravaginal devices for controlled delivery of antiprotozoal agents in the treatment and prevention of sexually transmitted infections.
This study presents clinical observations in pregnancy made on aborigines of the deep jungle and "outside" populations. Migration out of the jungle results in lowered nutritional status as a result of low socio-economic status in the "outside" aborigine. This, together with food habits, increased family size and higher incidence of helminthic infestations, results in lower mean values of Hb, PVC and MCHC and a higher prevalence of anaemia in pregnancy in the migrant aborigine. A higher population density in the "outside" population resulting in frequent intermingling and increased chances of cross-contamination probably explains the increased vaginal bacterial growth in the "outside" Aborigine women. A higher prevalence of vaginal candidiasis in the "outside" aborigine woman is probably related to exposure to oral contraceptives and broad-spectrum antibiotics. On the other hand, better medical and obstetrical services become more readily available to the "outside" aborigine and this results in a favourable influence on perinatal health.
Citation: Prevalence survey of sexually transmitted diseases among sex workers and women attending antenatal clinics: Malaysia (1999-2000). Manila: World Health Organization, Regional Office for the Western Pacific; 2001
The prevalence of sexually transmitted diseases (STD) among female drug abusers was studied by screening 130 new inmates of a rehabilitation centre. Data was collected using a structured questionnaire, physical examination and specimen collection for laboratory investigation at the Community Medicine Clinic of HUSM. The majority (64.6%) were Malays and self-confessed sex workers (77.7%). A high prevalence of syphilis (50.8%), hepatitis B (52.2%), moniliasis (23.8%), trichomoniasis (19.2%) and gonococcal vaginitis (8.5%) was noted. More than half of them harbour 2 or more STDs. Six subjects, of whom 5 were Malays, were HIV positive. The evidence indicates that female drug abusers need a thorough screening for STD followed by an aggressive treatment regimen. Since the majority of them were born Muslims, they should have access to spiritual counselling and rehabilitation, stressing on repentance, and adopting a compassionate and forgiving approach.