scholarly journals The ProtozoanTrichomonas vaginalisTargets Bacteria with Laterally Acquired NlpC/P60 Peptidoglycan Hydrolases

mBio ◽  
2018 ◽  
Vol 9 (6) ◽  
Author(s):  
Jully Pinheiro ◽  
Jacob Biboy ◽  
Waldemar Vollmer ◽  
Robert P. Hirt ◽  
Jeremy R. Keown ◽  
...  

ABSTRACTThe human eukaryotic pathogenTrichomonas vaginaliscauses trichomoniasis, a prevalent sexually transmitted infection. This extracellular protozoan is intimately associated with the human vaginal mucosa and microbiota, but key aspects of the complex interactions between the parasite and the vaginal bacteria remain elusive. We report thatT. vaginalishas acquired, by lateral gene transfer from bacteria, genes encoding peptidoglycan hydrolases of the NlpC/P60 family. Two of theT. vaginalisenzymes were active against bacterial peptidoglycan, retaining the active-site fold and specificity asdl-endopeptidases. The endogenous NlpC/P60 genes are transcriptionally upregulated inT. vaginalisin the presence of bacteria. The overexpression of an exogenous copy enables the parasite to outcompete bacteria from mixed cultures, consistent with the biochemical activity of the enzyme. Our study results highlight the relevance of the interactions of this eukaryotic pathogen with bacteria, a poorly understood aspect of the biology of this important human parasite.IMPORTANCETrichomonas vaginalisis a parasitic protozoan of the human urogenital tract that causes trichomoniasis, a very common sexually transmitted disease. Despite residing extracellularly and in close association with the vaginal bacteria (i.e., the microbiota), very little is known about the nature of the parasite-bacterium interactions. Our study showed that this parasite had acquired genes from bacteria which retained their original function. They produce active enzymes capable of degrading peptidoglycan, a unique polymer of the bacterial cell envelope, helping the parasite to outcompete bacteria in mixed cultures. This study was the first to show that a laterally acquired group of genes enables a eukaryotic mucosal pathogen to control bacterial population. We highlight the importance of understanding the interactions between pathogens and microbiota, as the outcomes of these interactions are increasingly understood to have important implications on health and disease.

2018 ◽  
Author(s):  
Jully Pinheiro ◽  
Jacob Biboy ◽  
Waldemar Vollmer ◽  
Robert P. Hirt ◽  
Jeremy R. Keown ◽  
...  

AbstractTrichomonas vaginalisis a human eukaryotic pathogen and the causative agent of trichomoniasis, the most prevalent non-viral sexually transmitted infection worldwide. This extracellular protozoan parasite is intimately associated with the human vaginal mucosa and microbiota but key aspects of the complex interactions between the parasite and the vaginal bacteria remain elusive. We report thatT. vaginalishas acquired, by lateral gene transfer from bacteria, genes encoding peptidoglycan hydrolases of the NlpC/P60 family. Two of theT. vaginalisenzymes were active against bacterial peptidoglycan, retaining the active site fold and specificity as DL-endopeptidases. The endogenous NlpC/P60 genes are transcriptionally up regulated inT. vaginaliswhen in the presence of bacteria. The over-expression of an exogenous copy produces a remarkable phenotype where the parasite is capable of competing out bacteria from mixed cultures, consistent with the biochemical activity of the enzymein vitro. Our study highlights the relevance of the interactions of this eukaryotic pathogen with bacteria, a poorly understood aspect on the biology of this important human parasite.Author summaryTrichomonas vaginalisis a protozoan parasite that causes a very common sexually transmitted disease known as trichomoniasis. This extracellular parasite resides in the vagina where it is in close association with the mucosa and the local microbiota. Very little is known about the nature of the parasite-bacteria interactions. Here, we report that this parasite had acquired genes from bacteria which retained their original function producing active enzymes capable of degrading peptidoglycan, a polymer that is chemically unique to the cell envelope of bacteria. Our results indicate that these enzymes help the parasite compete out bacteria in mixed cultures. These observations suggest that these enzymes may be critical for the parasite to establish infection in the vagina, a body site that is densely colonised with bacteria. Our study further highlights the importance of understanding the interactions between pathogens and microbiota, as the outcomes of these interactions are increasingly understood to have important implications on health and disease.


2016 ◽  
Vol 55 (1) ◽  
pp. 321-325 ◽  
Author(s):  
Erik Munson ◽  
David Wenten ◽  
Sheila Jhansale ◽  
Mary Kay Schuknecht ◽  
Nicki Pantuso ◽  
...  

ABSTRACTOf 1,493 encounters of males at a sexually transmitted infection (STI) clinic in a community with a high prevalence of STI,Chlamydia trachomatiswas detected in 8.7% andNeisseria gonorrhoeaewas detected in 6.6%. AdditionalTrichomonas vaginalisandMycoplasma genitaliumscreening found 17.4% and 23.9% of the encounters, respectively, to be positive for STI. STI agents were detected in 13.7% of urine specimens; addition of pharyngeal and rectal collections to the analysis resulted in detection of STI agents in 19.0% and 23.9% of encounters, respectively. A total of 101 (23.8%) encounters of identified STI involved sole detection ofM. genitalium. Expansion of the STI analyte panel (includingM. genitalium) and additional specimen source sampling within a comprehensive STI screening program increase identification of male STI carriers.


2015 ◽  
Vol 59 (11) ◽  
pp. 6891-6903 ◽  
Author(s):  
Kuo-Yang Huang ◽  
Fu-Man Ku ◽  
Wei-Hung Cheng ◽  
Chi-Ching Lee ◽  
Po-Jung Huang ◽  
...  

ABSTRACTTrichomonas vaginaliscolonizes the human urogenital tract and causes trichomoniasis, the most common nonviral sexually transmitted disease. Currently, 5-nitroimidazoles are the only recommended drugs for treating trichomoniasis. However, increased resistance of the parasite to 5-nitroimidazoles has emerged as a highly problematic public health issue. Hence, it is essential to identify alternative chemotherapeutic agents against refractory trichomoniasis. Tetracycline (TET) is a broad-spectrum antibiotic with activity against several protozoan parasites, but the mode of action of TET in parasites remains poorly understood. Thein vitroeffect of TET on the growth ofT. vaginaliswas examined, and the mode of cell death was verified by various apoptosis-related assays. Next-generation sequencing-based RNA sequencing (RNA-seq) was employed to elucidate the transcriptome ofT. vaginalisin response to TET. We show that TET has a cytotoxic effect on both metronidazole (MTZ)-sensitive and -resistantT. vaginalisisolates, inducing some features resembling apoptosis. RNA-seq data reveal that TET significantly alters the transcriptome via activation of specific pathways, such as aminoacyl-tRNA synthetases and carbohydrate metabolism. Functional analyses demonstrate that TET disrupts the hydrogenosomal membrane potential and antioxidant system, which concomitantly elicits a metabolic shift toward glycolysis, suggesting that the hydrogenosomal function is impaired and triggers cell death. Collectively, we providein vitroevidence that TET is a potential alternative therapeutic choice for treating MTZ-resistantT. vaginalis. The in-depth transcriptomic signatures inT. vaginalisupon TET treatment presented here will shed light on the signaling pathways linking to cell death in amitochondriate organisms.


2017 ◽  
Vol 56 (2) ◽  
Author(s):  
Jane R. Schwebke ◽  
C. A. Gaydos ◽  
T. Davis ◽  
J. Marrazzo ◽  
D. Furgerson ◽  
...  

ABSTRACT Trichomoniasis is the most prevalent curable sexually transmitted disease (STD). It has been associated with preterm birth and the acquisition and transmission of HIV. Recently, nucleic acid amplification tests (NAAT) have been FDA cleared in the United States for detection of Trichomonas vaginalis in specimens from both women and men. This study reports the results of a multicenter study recently conducted using the Xpert TV (T. vaginalis) assay to test specimens from both men and women. On-demand results were available in as little as 40 min for positive specimens. A total of 1,867 women and 4,791 men were eligible for inclusion in the analysis. In women, the performance of the Xpert TV assay was compared to the patient infected status (PIS) derived from the results of InPouch TV broth culture and Aptima NAAT for T. vaginalis. The diagnostic sensitivities and specificities of the Xpert TV assay for the combined female specimens (urine samples, self-collected vaginal swabs, and endocervical swabs) ranged from 99.5 to 100% and 99.4 to 99.9%, respectively. For male urine samples, the diagnostic sensitivity and specificity were 97.2% and 99.9%, respectively, compared to PIS results derived from the results of broth culture for T. vaginalis and bidirectional gene sequencing of amplicons. Excellent performance characteristics were seen using both female and male specimens. The ease of using the Xpert TV assay should result in opportunities for enhanced screening for T. vaginalis in both men and women and, hopefully, improved control of this infection.


mBio ◽  
2013 ◽  
Vol 4 (2) ◽  
Author(s):  
Kristin N. Parent ◽  
Yuko Takagi ◽  
Giovanni Cardone ◽  
Norman H. Olson ◽  
Maria Ericsson ◽  
...  

ABSTRACTThe flagellated protozoanTrichomonas vaginalisis an obligate human genitourinary parasite and the most frequent cause of sexually transmitted disease worldwide. Most clinical isolates ofT. vaginalisare persistently infected with one or more double-stranded RNA (dsRNA) viruses from the genusTrichomonasvirus, familyTotiviridae, which appear to influence not only protozoan biology but also human disease. Here we describe the three-dimensional structure ofTrichomonas vaginalisvirus 1 (TVV1) virions, as determined by electron cryomicroscopy and icosahedral image reconstruction. The structure reveals aT= 1 capsid comprising 120 subunits, 60 in each of two nonequivalent positions, designated A and B, as previously observed for fungalTotiviridaefamily members. The putative protomer is identified as an asymmetric AB dimer consistent with either decamer or tetramer assembly intermediates. The capsid surface is notable for raised plateaus around the icosahedral 5-fold axes, with canyons connecting the 2- and 3-fold axes. Capsid-spanning channels at the 5-fold axes are unusually wide and may facilitate release of the viral genome, promoting dsRNA-dependent immunoinflammatory responses, as recently shown upon the exposure of human cervicovaginal epithelial cells to either TVV-infectedT. vaginalisor purified TVV1 virions. Despite extensive sequence divergence, conservative features of the capsid reveal a helix-rich fold probably derived from an ancestor shared with fungalTotiviridaefamily members. Also notable are mass spectrometry results assessing the virion proteins as a complement to structure determination, which suggest that translation of the TVV1 RNA-dependent RNA polymerase in fusion with its capsid protein involves −2, and not +1, ribosomal frameshifting, an uncommonly found mechanism to date.IMPORTANCETrichomonas vaginaliscauses ~250 million new cases of sexually transmitted disease each year worldwide and is associated with serious complications, including premature birth and increased transmission of other pathogens, including HIV. It is an extracellular parasite that, in turn, commonly hosts infections with double-stranded RNA (dsRNA) viruses, trichomonasviruses, which appear to exacerbate disease through signaling of immunoinflammatory responses by human epithelial cells. Here we report the first three-dimensional structure of a trichomonasvirus, which is also the first such structure of any protozoan dsRNA virus; show that it has unusually wide channels at the capsid vertices, with potential for releasing the viral genome and promoting dsRNA-dependent responses by human cells; and provide evidence that it uses −2 ribosomal frameshifting, an uncommon mechanism, to translate its RNA polymerase in fusion with its capsid protein. These findings provide both mechanistic and translational insights concerning the role of trichomonasviruses in aggravating disease attributable toT. vaginalis.


mSphere ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Sally D. Warring ◽  
Frances Blow ◽  
Grace Avecilla ◽  
Jordan C. Orosco ◽  
Steven A. Sullivan ◽  
...  

ABSTRACT Trichomonas vaginalis is the causative agent of trichomoniasis, the most prevalent nonviral sexually transmitted infection worldwide. Repetitive elements, including transposable elements (TEs) and virally derived repeats, comprise more than half of the ∼160-Mb T. vaginalis genome. An intriguing question is how the parasite controls its potentially lethal complement of mobile elements, which can disrupt transcription of protein-coding genes and genome functions. In this study, we generated high-throughput RNA sequencing (RNA-Seq) and small RNA-Seq data sets in triplicate for the T. vaginalis G3 reference strain and characterized the mRNA and small RNA populations and their mapping patterns along all six chromosomes. Mapping the RNA-Seq transcripts to the genome revealed that the majority of genes predicted within repetitive elements are not expressed. Interestingly, we identified a novel species of small RNA that maps bidirectionally along the chromosomes and is correlated with reduced protein-coding gene expression and reduced RNA-Seq coverage in repetitive elements. This novel small RNA family may play a regulatory role in gene and repetitive element expression. Our results identify a possible small RNA pathway mechanism by which the parasite regulates expression of genes and TEs and raise intriguing questions as to the role repeats may play in shaping T. vaginalis genome evolution and the diversity of small RNA pathways in general. IMPORTANCE Trichomoniasis, caused by the protozoan Trichomonas vaginalis, is the most common nonviral sexually transmitted infection in humans. The millions of cases each year have sequelae that may include complications during pregnancy and increased risk of HIV infection. Given its evident success in this niche, it is paradoxical that T. vaginalis harbors in its genome thousands of transposable elements that have the potential to be extremely detrimental to normal genomic function. In many organisms, transposon expression is regulated by the activity of endogenously expressed short (∼21 to 35 nucleotides [nt]) small RNA molecules that effect gene silencing by targeting mRNAs for degradation or by recruiting epigenetic silencing machinery to locations in the genome. Our research has identified small RNA molecules correlated with reduced expression of T. vaginalis genes and transposons. This suggests that a small RNA pathway is a major contributor to gene expression patterns in the parasite and opens up new avenues for investigation into small RNA biogenesis, function, and diversity.


2018 ◽  
Vol 86 (8) ◽  
Author(s):  
Niha Phukan ◽  
Anna E. S. Brooks ◽  
Augusto Simoes-Barbosa

ABSTRACTTrichomoniasis, a prevalent sexually transmitted infection, is commonly symptomatic in women. The causative agent isTrichomonas vaginalis, an extracellular protozoan parasite. The host-protective mechanisms and molecules of vaginal lactobacilli that counteract this pathogen are largely unknown. This study examines the inhibition promoted byLactobacillus gasseriagainst the adhesion ofT. vaginalisto host cells, a critical virulence aspect of this pathogen. We observed that the vaginal strainL. gasseriATCC 9857 is highly inhibitory by various contact-dependent mechanisms and that surface proteins are largely responsible for this inhibitory phenotype. We found that the aggregation-promoting factor APF-2 from these bacteria significantly contributes to inhibition of the adhesion ofT. vaginalisto human vaginal ectocervical cells. Understanding the molecules and mechanisms used by lactobacilli to protect the host againstT. vaginalismight help in the development of novel and specific therapeutic strategies that take advantage of the natural microbiota.


Planta Medica ◽  
2021 ◽  
Author(s):  
Nathalya Tesch Brazil ◽  
Bruna Medeiros-Neves ◽  
Flávia Nathiely Silveira Fachel ◽  
Vanessa Pittol ◽  
Roselena Silvestri Schuh ◽  
...  

Abstract Trichomonas vaginalis causes trichomoniasis, a nonviral sexually transmitted infection with a high prevalence worldwide. Oral metronidazole is the drug of choice for the treatment of this disease, although high levels of T. vaginalis resistance to this agent are well documented in the literature. This study describes the anti-T. vaginalis activity of an optimized coumarin-rich extract from Pterocaulon balansae. Optimization was performed to maximize extraction of total coumarins by means of a 3-level Box-Behnken design, evaluating the effect of three factors: extraction time, plant : solvent ratio, and ethanol concentration. Optimum conditions were found to be 5 h extraction time and a plant : solvent ratio of 1% (w/v) and 60% (v/v) ethanol, which resulted in approximately 30 mg of total coumarins/g of dry plant. The coumarin-enriched extract exhibited a minimum inhibitory concentration of 30 µg/mL and an IC50 of 3.2 µg/mL against T. vaginalis, a low cytotoxicity, and a high selectivity index (18 for vaginal epithelial cells and 16 for erythrocytes). The coumarins permeation/retention profile through porcine vaginal mucosa was evaluated in Franz-type diffusion cells. After 8 h of kinetics, coumarins were detected in the tissue (4.93 µg/g) without detecting them in the receptor compartment. A significant increase of coumarins in the mucosa layers (8.18 µg/g) and receptor compartment (0.26 µg/g) was detected when a T. vaginalis suspension (2 × 105 trophozoites/mL) was previously added onto the mucosa. No alterations were visualized in the stratified squamous non-keratinized epithelium of the porcine vaginal mucosa after contact with the extract. Overall, these results suggest that the P. balansae coumarin-rich extract may have potential as a treatment for trichomoniasis.


Author(s):  
Shiren Ali Al Hamzawi

Estimates of Trichomonas vaginalis prevalence in pregnant women are variable with few studies in Iraq.T. vaginalis is a worldwide prevalent sexually transmitted infection,but fortunately,it is very treatable. Researchers believed that pregnancy is one of the effective factors for T. vaginalis infection in women.A cross-sectional study performed in Obstetrics and Gynecology Department at Maternity and Children Teaching Hospital in Al-Diwaniya city on two hundred female pregnant patients between the ages of 16-45 years. These females had no intercourse for 2–3 days,not using drugs (antibiotics,antiprotozoal or steroids) for the last 15 days. Vaginal discharges of any type with or without itching,burning sensation or both were their main complaints. Vaginal swabs were taken from all participating patients for direct wet mount microscopy and culture for the detection of Trichomonas vaginalis infection. The study showed that twelve out of two hundred examined pregnant women (6%) presented with T. vaginalis infection. The infection was more in those with mothers’ age (26-35) years,housewives,low education,higher parity,and of rural residents. Other maternal variables were not significantly associated with T. vaginalis infection. The study showed a prevalence of (6%) of T. vaginalis infection in pregnant female attendees. Infection was more in those with mothers ’age (26-35) years,housewives,low educational level,higher parity,and living in rural areas.


2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Martin Obermeier ◽  
Monia Pacenti ◽  
Robert Ehret ◽  
Francesco Onelia ◽  
Rory Gunson ◽  
...  

AbstractObjectivesAutomated molecular analyzers have accelerated diagnosis, allowing earlier intervention and better patient follow-up. A recently developed completely automated molecular analyzer, Alinity™ m (Abbott), offers consolidated, continuous, and random-access testing that may improve molecular laboratory workflow.MethodsAn international, multicenter study compared laboratory workflow metrics across various routine analyzers and Alinity m utilizing assays for human immunodeficiency virus type 1 (HIV-1), hepatitis C virus (HCV), hepatitis B virus (HBV), high-risk human papillomavirus (HR HPV), and sexually transmitted infection (STI) (Chlamydia trachomatis [CT]/Neisseria gonorrhoeae [NG]/Trichomonas vaginalis [TV]/Mycoplasma genitalium [MG]). Three turnaround times (TATs) were assessed: total TAT (sample arrival to result), sample onboard TAT (sample loading and test starting to result), and processing TAT (sample aspiration to result).ResultsTotal TAT was reduced from days with routine analyzers to hours with Alinity m, independent of requested assays. Sample onboard TATs for standard workflow using routine analyzers ranged from 7 to 32.5 h compared to 2.75–6 h for Alinity m. The mean sample onboard TAT for STAT samples on Alinity m was 2.36 h (±0.19 h). Processing TATs for Alinity m were independent of the combination of assays, with 100% of results reported within 117 min.ConclusionsThe consolidated, continuous, random-access workflow of Alinity m reduces TATs across various assays and is expected to improve both laboratory operational efficiency and patient care.


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