Search

Background: Trichomonas vaginalis (T. vaginalis) is an anaerobic flagellated parasite that usually infects the female genitourinary system. The traditional diagnostic tools of T. vaginalis, like wet-mount microscopy, have several limitations regarding technical requirements and accuracy. Consequently, to control the spread of trichomoniasis, fast and accurate detection of trichomoniasis represents a crucial factor in the treatment plan. Objectives: This study was performed to utilize polymerase chain reaction (PCR) assays as a diagnostic tool for T. vaginalis. Materials and Methods: Vaginal swabs were obtained from 186 women who were admitted to the gynecology clinic in Al Sadiq Hospital in Babil Governorate in Iraq. Additionally, a questionnaire was gathered from the female participants regarding their age, residency, history of abortion, and symptoms. DNAs were extracted from the swabs, and conserved β-tubulin genes were amplified using specific primers. Results: Of the 186 participant females, 40 women showed positive PCR outcomes of T. vaginalis β-tubulin gene versus only seven positive cases detected by wet-mount microscopy. Residency and age-related prevalence of T. vaginalis infection were not statistically significant, despite the higher positive cases reported in urban areas. Additionally, higher percentages of history of abortion were seen in infected females versus noninfected females. Clinical diagnosis demonstrated that vaginal discharge and itching were the most prevalent symptoms reported by infected females. Conclusion: PCR assay used in the current study showed a beneficial and fast diagnostic ability of T. vaginalis because of its high sensitivity and specificity. Consequently, this would blunt the transmission of trichomoniasis among individuals, especially infected patients with asymptomatic properties.

Copyright of Medical Journal of Babylon is the property of Wolters Kluwer India Pvt Ltd and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Trichomonas vaginalis is the etiologic agent of trichomoniasis, a worldwide distributed sexually transmitted infection (STI) that affects the genitourinary tract. Even though this disease already has a treatment in the prescription of drugs of the 5-nitroimidazole class, described low treatments adhesion, adverse side effects and cases of resistant isolates demonstrate the need for new formulations. With this in mind, chalcones emerge as a potential alternative to be tested, being compounds widely distributed in nature, easy to chemically synthesize and presenting several biological activities already reported. In this experiment, we evaluated the antiparasitic activity of 10 chalcone at a concentration of 100 μM against ATCC 30236 T. vaginalis isolates, considering negative (live trophozoites), positive (Metronidazole 100 μM) and vehicle (DMSO 0.6%) controls. Compounds 3a, 3c, 3 g and 3i showed promising results, with MICs set at 70 μM, 80 μM, 90 μM and 90 μM, respectively (p < 0,05). Cytotoxicity assays were performed on VERO and HMVII cell lines and revealed low inhibition rates at concentrations bellow 20 μM. To elucidate a possible mechanism of action for these molecules, the DPPH, ABTS and FRAP assays were performed, in which none of the four compounds presented antioxidant activity. Assays to verify ROS and lipid peroxidation in the parasite membrane were performed. None of the tested compounds identified ROS accumulation after incubation with trophozoites. 3 g molecule promoted an increase in MDA production after incubation. Results presented in this paper demonstrate the promising trichomonicidal profile, although further tests are still needed to optimize their performance and better elucidate the mechanisms of action involved. [Display omitted] • Chalcone analogue compounds showed antiparasitic activity against T. vaginalis. • Cytotoxicity assays revealed low inhibition rates at concentrations below 20 μM. • No antioxidant activity was detected in the tested compounds. • The enzymes TvLDH and TvPNP showed better performance in molecular docking. [ABSTRACT FROM AUTHOR]

Trichomonas vaginalis is a unicellular flagellated protozoan that resides in female and male genital tract and considered the most prevalent sexually transmitted infectious parasite. The infection rate is relatively equivalent between male and female but trichomoniasis is usually asymptomatic in men. Primary triggering of host inflammatory response to this parasite is not fully understood and most studies address the local reaction of the parasite in female genital tract. In this study, two pro-inflammatory cytokines, IL-8 and IL-12, were investigated in the serum of infected women with Trichomonas vaginalis during acute and chronic stages of the disease. The results demonstrated that the level of IL-8 was significantly higher along the acute and chronic disease stages in female patients. However, the level of IL-12 was significantly increased in the acute group but exceptionally decreased in the chronic group of patients, in comparison with the control group. These results indicate that local infection of the vaginal Trichomoniasis may alter the levels of the studied inflammatory cytokines during infection. [ABSTRACT FROM AUTHOR]

Trichomoniasis, caused by the protozoan Trichomonas vaginalis, is the most prevalent non-viral sexually transmitted infection that affects over 170 million people worldwide. The only type of drug recommended for the therapeutic control of trichomoniasis is the 5-nitroimidazoles, although there have been reports of some undesirable side effects and clinical resistance. Hence, the need for the search for new tricomonicidal agents is necessary. In a previous work, we demonstrated that two 2-amino-4-aryl thiazole derivatives (ATZ-1 and ATZ-2) possess a portent antigiardial effect. In the current paper, we investigated the in vitro antitrichomonal activity of these thiazole compounds. Both ATZ-1 and ATZ-2 reduced the viability and growth of parasites in a dose-dependent manner, with an IC50 value of 0.15 μg/mL and 0.18 μg/mL, respectively. Furthermore, both thiazole compounds were able to decrease the proteolytic activity in T. vaginalis trophozoites compared with untreated parasites. Interestingly, a full proteolytic inhibition profile was observed in the 50-kDa region which was associated with the decreased expression of the gene that codes for the trichomonad protease TvMP50. The docking simulations predicted strong interactions of the thiazole compounds in the TvMP50 protease's active site, suggesting a possible role as protease inhibitors. Our results demonstrate the potential of 2-amino-4-aryl thiazole derivatives as trichomonicidal compounds and could be, mechanistically, involved in the inhibition of key trichomonad proteases. [ABSTRACT FROM AUTHOR]

The protozoan parasite Trichomonas vaginalis (TV), exclusively adapted to the human genital tract, is one of the most common sexually transmitted pathogens. Adding to the complexity of the host-pathogen interactions, the parasite harbors TV-specific endosymbiont viruses (Trichomonasvirus , TVV). It was reported that small extracellular vesicles (sEVs) released by TV play a role in host immunity; however, the role of the viral endosymbiosis in this process remained unknown. We hypothesized that the virus may offer evolutionary benefit to its protozoan host at least in part by altering the immunomodulatory properties of sEVs spreading from the site of infection to non-infected immune effector cells. We infected human vaginal epithelial cells, the natural host of the parasite, with TV natively harboring TVV and an isogenic derivative of the parasite cured from the viral infection. sEVs were isolated from vaginal cell culture 24 h post TV infection and from medium where the isogenic TV strains were cultured in the absence of the human host. sEVs from TVV-negative but not TVV-positive parasites cultured alone caused NF-κB activation and increase of IL-8 and RANTES expression by uterine endocervical cells, which provide innate immune defense at the gate to the upper reproductive tract. Similarly, mononuclear leukocytes increased their IL-8, IL-6 and TNF-α output in response to sEVs from virus-negative, but not isogenic virus-positive parasites, the latter exosomes being immunosuppressive in comparison to TV medium control. The same phenomenon of suppressed immunity induced by the TVV-positive compared to TVV-negative phenotype was seen when stimulating the leukocytes with sEVs originating from infected vaginal cultures. In addition, the sEVs from the TVV-positive infection phenotype suppressed immune signaling of a toll-like receptor ligand derived from mycoplasma, another frequent TV symbiont. Quantitative comparative proteome analysis of the secreted sEVs from virus-positive versus virus-negative TV revealed differential expression of two functionally uncharacterized proteins and five proteins involved in Zn binding, protein binding, electron transfer, transferase and catalytic activities. These data support the concept that symbiosis with viruses may provide benefit to the protozoan parasite by exploiting sEVs as a vehicle for inter-cellular communications and modifying their protein cargo to suppress host immune activation. [ABSTRACT FROM AUTHOR]

Trichomonas infection is a scientific and clinical problem in dermatology, urology, obstetrics and gynecology. This infection in men and women is often causes to urethritis, cystitis, prostatitis, post-abortion infection, premature labor and other diseases. This literature review has shown that in spite of a lot of numbers of scientific data concerning the impact of trichomoniasis to urogenital bioceonosis of both men and women, there is no enough data about influence of trichomoniasis to urinary microecology in sexual partners. Meanwhile, this question is in the field of scientific and applied interest and need to be studied.

The currently available nucleic acid amplification tests (NAATs) for trichomoniasis are accurate, quick and confirmative with superior sensitivity than traditional culture-based microbiology assays. However, these assays are associated with problems of carry over contamination, false positive results, requirement of technical expertise for performance and detection of end product. Hence, a diagnostic assay with easy visualization of the amplified product will be profitable. An in-house, rapid, sensitive, specific molecular-beacon-based PCR assay, using primers against pfoB gene of Trichomonas vaginalis, was developed and evaluated using dry ectocervical swabs (n=392) from symptomatic females with vaginal discharge. Total DNA was isolated and used as template for the PCR assays. The performance and reproducibility of PCR assay was evaluated by composite reference standard (CRS). For easy visualization of the amplified product, molecular-beacon was designed and amplicons were visualized directly using fluorescent handheld dark reader or by Micro-Plate Reader. Molecular-beacons are single-stranded hairpin shaped nucleic acid probes composed of a stem, with fluorophore/quencher pair and a loop region complementary to the desired DNA. The beacon-based PCR assay designed in the present study is highly specific as confirmed by competition experiments and extremely sensitive with detection limit of 20 fg of genomic DNA (3–4 pathogens). The minimum infrastructure requirement and ease to perform the assay makes this method highly useful for resource poor countries for better disease management. [ABSTRACT FROM AUTHOR]

Prevalence of urinary parasitic infections with emphasis on T. vaginalis and S. haematobium among patients attending four selected hospitals in Gboko viz: Mkar hospital, Myom hospital, General hospital and Division hospital all in Gboko Local Government area of Benue State Nigeria, was investigated. A total number of 400 urine samples (100 from each hospital) were aseptically collected into sterile specimen bottles. Samples were immediately transferred to Lens Gabriel School of Medical Laboratory Sciences Mkar, Benue State for microscopic analyses and examination. Data were analysed and presented using both descriptive and inferential statistics. Based on age and marital status, results showed that prevalence rate of trichomoniasis and schistosomiasis were highest among (C) that is age group 21years and above. In all the hospitals, prevalence rate of T. vaginalis was highest with 12% followed by S. haematobium with an infection rate of 11% among the married patients as compared to the single patients. In the case of co infection, prevalence rate was also highest in the married patients (2.5%) as opposed to the single patients (1.5%). There was no significant level of associations between urinary tract infections, age and marital status of patients with 95% confidence limit. This work has therefore provided baseline information on the prevalence of these two infections in the four hospitals in Gboko Local Government Area. Thus, there is need for urgent effective control measures. [ABSTRACT FROM AUTHOR]

Trichomonas vaginalis is the cause of one of the most common types of vaginitis, trichomoniasis. Clinical manifestations of symptomatic women are generally nonspecific, but include vaginal discharge, vaginitis and irritation. T. vaginalis infection has also been linked to the increased risk of human immunodeficiency virus transmission. The aim of the study was to compare a real-time polymerase chain reaction (PCR) assay with culture and wet-mount examination for the detection of T. vaginalis. This is a descriptive analytical study. Vaginal swabs from 504 female patients with suspected vaginitis were included in the study. They were subjected to culture, wet-mount microscopic examination (WM), and real-time PCR. Real-time fluorescence resonance energy transfer hybridization probe PCR assays were used in this study to test for T. vaginalis DNA, targeting the b-tubulin genes. The result of the PCR was compared with culture and wet-mount microscopy. WM were positive for T. vaginalis in 60/504 cases (11.9%) and cultures were positive for T. vaginalis in 116/504 cases (23%). Real-time PCR was done on 50/504 specimens which was randomly chosen, 33/50 (66%) cases were positive for T. vaginalis (78% sensitivity), 25/50 (50%) of them were culture and wet-mount examination positive. Of them, 1/33 (3%) were culture positive and wet examination negative, 7/33 (21%) real-time positive were negative by culture while 17/50 (34%) real-time PCR negative cases were also negative by both techniques. The time taken for PCR assay was 4 to 8 h whereas positive protozoal cultures took up to 4 days. The Real-time PCR done in this study, provide a rapid, sensitive and specific diagnosis of T. vaginalis infection. [ABSTRACT FROM AUTHOR]

Trichomonas vaginalis is a sexually transmitted protozoan infection resulting in vulvo-vaginitis and altered vaginal discharge in symptomatic women. Trichomoniasis has been implicated in causing adverse pregnancy outcomes such as low birth weight and pre-term labour. Metronidazole is the recommended first-line treatment for trichomonal infection. Other nitroimidazoles, such as tinidazole, are used as alternative regimens with similar activity but at a greater expense. Treatment failure usually represents patient non-compliance or re-infection, although metronidazole resistance has previously been documented. Antimicrobial susceptibility testing for T. vaginalis is currently not available in the UK. Patients with disease unresponsive to first-line treatments pose a major challenge, as therapeutic options are limited. We present the case of a patient with presumed resistant infection during pregnancy, and the additional treatment issues that this presented. [ABSTRACT FROM AUTHOR]

• A nanobiosensor was designed for Trichomonas vaginalis. • Trichomonas vaginalis genosensor had detection limit of 31.06 pg mL−1 for the detection of genomic DNA sequences in spectrophotometric results. • Trichomonas vaginalis genosensor was evaluated on synthetic target, genome and clinical samples. Trichomoniasis, an infectious disease caused by a parasite called Trichomonas vaginalis (T. vaginalis), enhances the risk of HIV infection, cervical and prostate cancer, and infertility. Therefore, efforts have to be made for accurate, specific, and rapid diagnosise and treatment of trichomoniasis. Today, optical nanosensors have created an opportunity for diagnosis without sophisticated and expensive tools and the need for expertise; at the same time, they are highly sensitive and fast. An optical nano-genosensor was designed by conjugation of gold nanoparticles and a specific oligonucleotide (AuNPs-probe) from repeated DNA target for specific and sensitive polymerase chain reaction diagnosis of T. vaginalis gene sequence (L23861.1). The hybridization of AuNPs-probe was investigated with different concentrations of complementary sequence in synthesized target, gene sequence of standard T. vaginalis genomic DNA extraction, and PCR products of genomic DNA samples extracted from patients. Negative samples including synthesized non-complementary sequence, genomics DNA of other pathogens, and genomics DNA of healthy persons were considered for proof of the accuracy of the sensor function. The occurrence of correct hybridization was detected by adding acid to the medium and observing the changes in the color of the medium and spectroscopic spectrum. Based on spectrophotometric results, the fabricated genosensor had detection limits of 35.16 and 31 pg μL−1 for the detection of synthetic target and genomic DNA sequences, respectively. The results confirmed the correct function of genosensor for the detection of T. vaginalis in clinical samples. Advantages such as low cost, visual detection, speed, and easy diagnosis encourage the use of this sensor in pathogen detection in the future. [ABSTRACT FROM AUTHOR]

Adherence to the surface of the host cell is the precondition for T. vaginalis parasitism and pathogenicity, causing urogenital infection. The AP65 of T. vaginalis (TvAP65) involves in the process of adhesion. So, the present study was aimed at investigating the molecular characterization and vaccine candidacy of TvAP65 for protecting the host from the onset of Trichomoniasis. The open reading frame (ORF) of TvAP65 was amplified and then inserted into pET-32a (+) to clone recombinant TvAP65 (rTvAP65). The immunoblotting determined the immunogenicity and molecular size of TvAP65, while immunofluorescence staining visualized and the precise localization of TvAP65 in T. vaginalis trophozoites. Animal challenge and enzyme-linked immunosorbent assay (ELISA) test were used to evaluate the immunoprotection and the types of the immune response of TvAP65. By the sequence analysis, TvAP65 encoded a 63.13 kDa protein that consisted 567 amino acid residues with a high antigenic index. The western blotting revealed that rTvAP65 and native TvAP65 could interact with the antibodies in the rat serums post hoc rTvAP65 immunization and the serums from the mice that were experimentally infected with T. vaginalis , respectively. Immunofluorescence stained TvAP65 on the surface of T. vaginalis trophozoites. Moreover, following emulsification with Freund's adjuvant, rTvAP65 was subsequently administered to BALB/c mice three times at 0, 2, and 4 weeks and the results from this animal challenge experiments showed significant increases in immunoglobulins of IgG2a, IgG1, and IgG, and cytokine of IFN-γ, and IL-2, and 10. Lastly, rTvAP65 vaccinated animals had a prolonged survival time (26.80 ± 4.05) after challenged by T. vaginalis. TvAP65 mediated the adhesion of T. vaginalis to the host epithelia for the pathogenesis of the parasite and can be considered as a candidate protein for designing a functional vaccine that induces cell-mediated and humoral immunity against the T. vaginalis infection. Image 1 • Molecular Cloning and sequence analysis of the TvAP65 were completed. • TvAP65 was located on the surface of the trophozoites of T. vaginalis and could stimulate the production of specific antibodies. • Recombinant TvAP65 protein was capable of inducing partial protective effects in mice challenged with parasites. [ABSTRACT FROM AUTHOR]