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.

UNASSIGNED: Trichomonas vaginalis is a sexually transmitted protozoan parasite that usually causes infections in women. Metronidazole is used as the first choice in the treatment of this parasitic disease, but there is a need for new drugs since 1980\'s with increasing numbers of reported resistance. In this study, it was aimed to determine the antitrichomonal activity of the major components of Cinnamomum zeylanicum (cinnamon) and Thymus vulgaris (thyme) essential oils, cinnamaldehyde, carvacrol and thymol against metronidazole resistant and susceptible T. vaginalis strains, and to determine their interaction with metronidazole by checkerboard method.
UNASSIGNED: Cinnamaldehyde, carvacrol, thymol and metronidazole were obtained commercially. Two clinical isolates and one metronidazole resistant T. vaginalis reference strain were used in the study. MIC50 and MLC values of essential oil components and metronidazole were determined by broth microdilution method. The combinations of essential oil components with metronidazole were determined by the checkerboard method.
UNASSIGNED: According to in vitro activity tests, cinnamaldehyde was determined to be most effective essential oil component. Clinical isolates were susceptible to metronidazole. In combination study, metronidazole showed synergy with cinnamaldehyde and carvacrol, and partial synergy with thymol.
UNASSIGNED: It was determined that cinnamaldehyde, carvacrol and thymol, which are known to have high antimicrobial activity, also have strong activity against T. vaginalis isolates and show a synergistic interaction with metronidazole. The use of metronidazole at lower doses in the synergistic interaction may contribute to the literature in terms of reducing drug side effects, creating a versatile antimicrobial target, and reducing the rate of resistance development.
UNASSIGNED: Trichomonas vaginalis genellikle kadınlarda enfeksiyona neden olan ve cinsel yolla bulaşan bir protozoon parazittir. Parazitin neden olduğu hastalığın tedavisinde ilk tercih olarak metronidazol kullanılmaktadır. Ancak 1980 yılından sonra artan sayılarda direnç gelişiminin rapor edilmesi ile yeni ilaç arayışlarına ihtiyaç duyulmuştur. Bu çalışmada, Cinnamomum zeylanicum (tarçın) ve Thymus vulgaris (kekik) uçucu yağlarının majör bileşenleri olan sinnamaldehit, karvakrol ve timolün metronidazole dirençli ve duyarlı T. vaginalis izolatlarına karşı anti-trichomonal etkinliğinin belirlenmesi ve metronidazol ile etkileşiminin checkerboard (dama tahtası) yöntemi ile gösterilmesi amaçlandı.
UNASSIGNED: Çalışmada kullanılan sinnamaldehit, karvakrol, timol ve metronidazolün saf formları ticari olarak temin edildi. Çalışmada, iki klinik izolat ve bir adet metronidazole dirençli T. vaginalis standart (ATCC 50143) suşu kullanıldı. Uçucu yağ bileşenlerinin ve metronidazolün MIK50 ve MLK (minimum letal konsantrasyonu) değerleri sıvı mikrodilüsyon yöntemi, metronidazol ile kombinasyonu ise checkerboard (dama tahtası) yöntemi ile saptandı.
UNASSIGNED: İn vitro etkinlik testlerine göre, en etkili uçucu yağ bileşeninin sinnamaldehit olduğu belirlendi. Klinik izolatların metronidazole duyarlı olduğu saptandı. Checkerboard yöntemi ile yapılan kombinasyon çalışması değerlendirildiğinde, sinnamaldehit ve karvakrolün metronidazol ile kombinasyonunda sinerji, timolün metronidazol ile kombinasyonunda ise kısmi sinerji görüldü.
UNASSIGNED: Yüksek antimikrobiyal aktiviteye sahip olduğu bilinen sinnamaldehit, karvakrol ve timol’ün T. vaginalis izolatlarına karşı güçlü aktiviteye sahip olduğu ve metronidazol ile sinerjistik etkileşim gösterdiği belirlendi. Sinerjik etkileşimde metronidazolün daha düşük dozlarda kullanılması ilaç yan etkilerinin azaltılması, çok yönlü bir antimikrobiyal hedef oluşturulması ve direnç gelişme hızının düşürülmesi açısından literatüre katkı sağlayabilir.

UNASSIGNED: Preterm delivery is a common complication during pregnancy periods and imposes a high cost on the healthcare system due to the care needs of premature babies. Sexually transmitted infections are one of the effective factors in the occurrence of preterm delivery and the diagnosis and treatment of these infections are effective in reducing complications and preventing preterm delivery. In this study, the role of Trichomonas vaginalis (T. vaginalis [TV]) infection in preterm delivery has been evaluated.
UNASSIGNED: In a prospective case-control study, women with preterm birth were assigned to the case group, and women with full-term delivery on the same day were also assigned randomly to the control group. After receiving the history and physical examination, a sample was taken from the cervix for TV culture. The data were included in the SPSS version 23 software. A significance level of less than 0.05 was considered.
UNASSIGNED: The overall prevalence of this infection was 10%. The prevalence of chlamydial infection was 2% among mothers with full-term delivery and 16.4% among mothers with premature birth, and there was a significant difference between the two groups (P = 0.021). The logistic regression analysis to determine the effect of Trichomonas infection on premature birth showed that there was the probability of the occurrence of premature delivery increases in mothers with trichomoniasis infection with lower age, higher body mass index, the presence of underlying disease, lower educational level, housewives, lower parity and gravity and having a history of fetus abortion more than 13 times with its occurrence probability occurs in mothers without Trichomonas infection (P = 0.046, Exp (β) =13.266).
UNASSIGNED: According to the present results, TV screening for pregnant women, especially in high-risk groups, is emphasized to reduce the incidence of preterm delivery and related complications, especially neonatal complications.

Trichomonas vaginalis (T. vaginalis) is a widespread and important sexually transmitted pathogen. Adherence to the surface of the host cell is the precondition forthis parasite\'s parasitism and pathogenicity. Adhesion protein 65 (TvAP65) plays a key role in the process of adhesion. However, how TvAP65 mediates the adhesion and pathogenicity of T. vaginalis to host cellsis unclear. In this study, we knocked down the expression of TvAP65 in trophozoites by small RNA interference. The number of T. vaginalis trophozoites adhering to VK2/E6E7 cells was decreased significantly, and the inhibition of VK2/E6E7 cells proliferation and VK2/E6E7 cells apoptosis and death induced by T. vaginalis were reduced, after the expression of TvAP65 was knocked down. Animal challenge experiments showed that the pathogenicity of trophozoites was decreased by passive immunization with anti-rTvAP65 PcAbs or blocking the TvAP65 protein. Immunofluorescence analysis showed that TvAP65 could bind to VK2/E6E7 cells. In order to screen the molecules interacting with TvAP65 on the host cells, we successfully constructed the cDNA library of VK2/E6E7 cells, and thirteen protein molecules interacting with TvAP65 were screened by yeast two-hybrid system. The interaction between TvAP65 and BNIP3 was further confirmed by coimmunoprecipitation and colocalization. When both TvAP65 and BNIP3 were knocked down by small RNA interference, the number of T. vaginalis adhering to VK2/E6E7 cells and the inhibition of VK2/E6E7 cells proliferation were significantly lower than those of the group with knockdown of TvAP65 or BNIP3 alone. Therefore, the interaction of TvAP65 and BNIP3 in the pathogenesis of T. vaginalis infecting host cells is not unique and involves other molecules. Our study elucidated that the interaction between TvAP65 and BNIP3 mediated the adhesion and pathogenicity of T. vaginalis to host cells, provided a basis for searching for the drug targets of anti-T. vaginalis, and afforded new ideas for the prevention and treatment of trichomoniasis.

BACKGROUND: Trichomonas vaginalis is a widespread and important sexually transmitted pathogen. Adherence to the surface of the host cell is the precondition for the parasitism and pathogenicity of this parasite. Trichomonas vaginalis adhesion protein 33 (TvAP33) plays a key role in the process of adhesion, but how this protein mediates the adhesion and pathogenicity of T. vaginalis to host cells is unclear.
METHODS: The expression of TvAP33 in trophozoites was knocked down by small interfering RNA. VK2/E6E7 cells and mice infected with T. vaginalis were used to evaluate the pathogenicity of T. vaginalis. We constructed a complementary DNA library of VK2/E6E7 cells and screened the protein molecules interacting with TvAP33 by the yeast two-hybrid system. The interaction between TvAP33 and BNIP3 (Bcl-2 interacting protein 3) was analyzed by co-immunoprecipitation and colocalization.
RESULTS: Following knockdown of TvAP33 expression, the number of T. vaginalis trophozoites adhering to VK2/E6E7 cells decreased significantly, and the inhibition of VK2/E6E7 cell proliferation and VK2/E6E7 cell apoptosis and death induced by T. vaginalis were reduced. Animal challenge experiments showed that the pathogenicity of trophozoites decreased following passive immunization with TvAP33 antiserum or blocking of the TvAP33 protein. Immunofluorescence analysis revealed that TvAP33 could bind to VK2/E6E7 cells. Eighteen protein molecules interacting with TvAP33 were identified by the yeast two-hybrid system. The interaction between TvAP33 and BNIP3 was further confirmed by co-immunoprecipitation and colocalization. When the expression of both TvAP33 and BNIP3 in trophozoites was knocked down by small RNA interference, the number of T. vaginalis adhering to VK2/E6E7 cells and the inhibition of VK2/E6E7 cell proliferation were significantly lower compared to trophozoites with only knockdown of TvAP33 or only BNIP3. Therefore, the interaction of TvAP33 and BNIP3 in the pathogenesis of T. vaginalis infecting host cells is not unique and involves other molecules.
CONCLUSIONS: Our study showed that the interaction between TvAP33 and BNIP3 mediated the adhesion and pathogenicity of T. vaginalis to host cells, providing a basis for searching for drug targets for T. vaginalis as well as new ideas for the prevention and treatment of trichomoniasis.

The mucosa of the female reproductive tract must reconcile the presence of commensal microbiota and the transit of exogenous spermatozoa with the elimination of sexually transmitted pathogens. In the vagina, neutrophils are the principal cellular arm of innate immunity and constitute the first line of protection in response to infections or injury. Neutrophils are absent from the vaginal lumen during the ovulatory phase, probably to allow sperm to fertilize; however, the mechanisms that regulate neutrophil influx to the vagina in response to aggressions remain controversial. We have used mouse inseminations and infections of Neisseria gonorrhoeae, Candida albicans, Trichomonas vaginalis, and HSV-2 models. We demonstrate that neutrophil infiltration of the vaginal mucosa is distinctively contingent on the ovarian cycle phase and independent of the sperm and pathogen challenge, probably to prevent sperm from being attacked by neutrophils. Neutrophils extravasation is a multi-step cascade of events, which includes their adhesion through selectins (E, P and L) and integrins of the endothelial cells. We have discovered that cervical endothelial cells expressed selectin-E (SELE, CD62E) to favor neutrophils recruitment and estradiol down-regulated SELE expression during ovulation, which impaired neutrophil transendothelial migration and orchestrated sperm tolerance. Progesterone up-regulated SELE to restore surveillance after ovulation.

Trichomonas vaginalis (T. vaginalis) infection is the most common non-viral sexually transmitted disease (STD) in the world. It can cause male reproductive dysfunction and infertility. However, the pathogenic mechanism is not clear. In this study, the excretory secretory proteins of T. vaginalis (TvESPs) were collected, concentrated, and sterilized. After sperm co-cultured with TvESPs, the survival rate and motility of sperms were analyzed by seminal routine examination, and the results showed that the TvESPs could significantly reduce the survival rate and motility of sperms. Fluorescence staining displayed that TvESPs could destroy the integrity of sperm acrosomes. Flow cytometry indicated that TvESPs induced sperm apoptosis. By mouse in vitro fertilization, we confirmed that TvESPs could significantly reduce the fertilization ability of sperms and negatively affect the development of the fertilized ovum. Via semi-quantitative analysis, we found that the apoptosis-related p27, SMAC, p53, BAX, BCL-2, XIAP, and BCL-W molecules were down-regulated in mouse sperm cells after interaction between the sperms and TvESPs, which played an important role in regulating sperm apoptosis. In conclusion, our study showed that T. vaginalis degraded semen quality and negatively affected male fertility by TvESPs. TvESPs may damage sperms by breaking the balance between sperm pro-apoptotic and anti-apoptotic molecules. This study proves that T. vaginalis infection is a risk factor for infertility.

Trichomonas vaginalis is a protozoan parasite with unicellular, flagellate, and anaerobic metabolism. It is the second most prevalent pathogen among sexually transmitted agents after viruses. Microscopic examination, culture, and molecular methods are used in the laboratory diagnosis of T. vaginalis. However, in most routine microbiology laboratories, microscopy is preferred instead of culture and molecular methods for T. vaginalis diagnosis because microscopy is cheaper than other methods. This study aimed to produce T. vaginalis in media that can be detected frequently in microbiology laboratories.
In this study, four media, namely, thioglucholate medium (THIO), brain heart infusion medium (BHI), tryptic soy broth medium (TSB), and Brucella broth medium (BRB) were modified and tested. Trypticase-yeast extract-maltose (TYM) medium was used as a reference medium. Each medium tested was enriched with three different serum additives. T. vaginalis trophozoite at a density of 104 parasites/mL was inoculated into each medium and incubated at 37 °C for 10 days. We determined the number of trophozoites using a hemocytometer, and the viability rates were determined using trypan blue.
Trichomonas vaginalis grew extremely well on THIO, BHI, and TSB media but not on BRB media. The time and number of parasites peaked were determined as 100×104 parasites/mL on THIO-ATS and THIO-FCS media on days five and four, respectively, 100×104 parasites/mL on BHI-ATS on day three, 98×104 parasite/mL on BHI-FCS media on day five, 100×104 parasites/mL on TSB-ATS on day four, and 82×104 parasite/mL on TSB-FCS media on day seven. Compared with the reference medium, TYM, T. vaginalis trophozoites survived significantly longer in THIO, BHI, and TSB media.
The rich content of THIO, TSB, and BHI media, which are widely available in routine microbiology laboratories, may allow T. vaginalis growth.
Trichomonas vaginalis, tek hücreli ve kamçılı bir parazit olup, cinsel yolla bulaşan etkenler arasında virüslerden sonra en sık görülen ikinci patojendir. T. vaginalis’in laboratuvar tanısı, mikroskobik inceleme, kültür ve moleküler yöntemler ile yapılmaktadır. Ancak rutin mikrobiyoloji laboratuvarlarının çoğunda kültür ve moleküler yöntemlerin maliyetli olması nedeniyle sadece mikroskopi ile tanı konulmaya çalışılmaktadır. Bu çalışmada, mikrobiyoloji laboratuvarında yaygın olarak bulunan besiyerlerinde T. vaginalis’in üretilmesi amaçlanmıştır.
Bu çalışmada, tiyoglukolat besiyeri (THIO), brain heart infusion besiyeri (BHI), tryptic soya broth besiyeri (TSB) ve Brucella broth besiyeri (BRB) olmak üzere dört besiyeri modifiye edilerek denenmiştir. Referans besiyeri olarak trypticase-yeast- extract-maltose (TYM) besiyeri kullanılmıştır. Denenen her bir besiyeri, üç farklı serum katkısı ile zenginleştirilmiştir. Her bir besiyerine 104 parazit/mL T. vaginalis trofozoiti inoküle edilmiş ve 37 °C’de 10 gün inkübe edilmiştir. Trofozoit sayısı hemasitometre, canlılık oranları ise tripan mavisi kullanılarak belirlenmiştir.
Besiyerlerindeki T. vaginalis üremesi, referans besiyeri olan TYM ile kıyaslanmıştır. T. vaginalis, at serumu ve fetal sığır serumu ile zenginleştirilmiş THIO, BHI ve TSB besiyerlerinde çok iyi ürerken, BRB besiyerinde ürememiştir. Buna ek olarak, insan serumu ile zenginleştirilmiş besiyerlerinin hiçbirinde T. vaginalis üremesi görülmemiştir. Parazitin en yüksek seviyeye ulaştığı zaman ve sayı, THIO-ATS ve THIO-FCS besiyerleri için sırasıyla, beşinci ve dördüncü günde 100x104 parazit/mL; BHI-ATS besiyeri için üçüncü günde 100x104 parazit/mL; BHI-FCS besiyeri için beşinci günde 98x104 parazit/mL; TSB-ATS besiyeri için dördüncü günde 100x104 parazit/mL ve TSB-FCS besiyeri için ise yedinci günde 82x104 parazit/mL olarak belirlenmiştir. Referans besyeri olan TYM ile kıyaslandığında T. vaginalis trofozoitlerinin yaşam süresinin THIO, BHI ve TSB besiyerlerinde çok daha uzun olduğu saptanmıştır.
THIO-ATS, THIO-FCS, TSB-ATS, TSB-FCS, BHI-ATS ve BHI-FCS besiyerlerinin, T. vaginalis’in üretilebilmesi açısından TYM ile kıyaslanabilir olduğu saptanmıştır. Rutin mikrobiyoloji laboratuvarında yaygın olarak bulunan THIO, TSB ve BHI besiyerlerinin zengin içeriği T. vaginalis’in kültürüne imkan sağlayabilir.

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.

BACKGROUND: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.