Background Trichomonas vaginalis is the causative agent of human trichomoniasis, the most common non-viral sexually transmitted infection world-wide. Despite its prevalence, little is known about the genetic diversity and population structure of this haploid parasite due to the lack of appropriate tools. The development of a panel of microsatellite makers and SNPs from mining the parasite's genome sequence has paved the way to a global analysis of the genetic structure of the pathogen and association with clinical phenotypes. Methodology/Principal Findings Here we utilize a panel of T. vaginalis -specific genetic markers to genotype 235 isolates from Mexico, Chile, India, Australia, Papua New Guinea, Italy, Africa and the United States, including 19 clinical isolates recently collected from 270 women attending New York City sexually transmitted disease clinics. Using population genetic analysis, we show that T. vaginalis is a genetically diverse parasite with a unique population structure consisting of two types present in equal proportions world-wide. Parasites belonging to the two types (type 1 and type 2) differ significantly in the rate at which they harbor the T. vaginalis virus, a dsRNA virus implicated in parasite pathogenesis, and in their sensitivity to the widely-used drug, metronidazole. We also uncover evidence of genetic exchange, indicating a sexual life-cycle of the parasite despite an absence of morphologically-distinct sexual stages. Conclusions/Significance Our study represents the first robust and comprehensive evaluation of global T. vaginalis genetic diversity and population structure. Our identification of a unique two-type structure, and the clinically relevant phenotypes associated with them, provides a new dimension for understanding T. vaginalis pathogenesis. In addition, our demonstration of the possibility of genetic exchange in the parasite has important implications for genetic research and control of the disease.

During oxidative stress, K63-linked polyubiquitin chains accumulate in the cell and modify a variety of proteins including ribosomes. Knowledge of the precise sites of K63 ubiquitination is key to understand its function during the response to stress. To identify the sites of K63 ubiquitin, we developed a new mass-spectrometry based method that quantified >1,100 K63 ubiquitination sites in yeast responding to oxidative stress induced by H2O2. We determined that under stress, K63 ubiquitin modified proteins are involved in several cellular functions including ion transport, protein trafficking, and translation. The most abundant ubiquitination sites localized to the head of the 40S subunit of the ribosome, modified assembled polysomes, and affected the binding of translation factors. The results suggested a new pathway of post-initiation control of translation during oxidative stress and illustrated the importance of high-resolution mapping of noncanonical ubiquitination events.

Trichomonas vaginalis is a parasite of the urogenital tract in men and women, with a worldwide presence and significant implications for global public health. T. vaginalis research entered the age of genomics with the publication of the first genome sequence in 2007, yet subsequent utilization of other ‘omics’ technologies and methods has been slow. Here, we review some of the tools and approaches available to interrogate T. vaginalis biology, with an emphasis on recent advances and current limitations, and draw attention to areas where further efforts are needed to effectively examine the complex and intriguing biology of the parasite.

suggests that phylotypes differ phenotypically. An accurate survey of genetic diversity and population structure will facilitate responsible selection of drug and/or vaccine targets for future treatments, and will enable better understanding of virulence factors contributing to the wide range of severity of symptoms associated with trichomoniasis. Objectives To develop a novel diagnostic protein in order to enhance early detection of syphilis infections and improve overall syphilis diagnosis. Methods Using pooled serum samples from patients infected with syphilis, immunoreactive regions of two previously identified diagnostic protein candidates, Tp0326 and Tp0453, were elucidated. Focusing on these regions, a chimeric protein construct was created for expression in Escherichia coli and expression conditions were optimised to produce soluble protein expression. This Tp0326/Tp0453 chimeric construct was screened against serum samples from; patients with primary, secondary, latent, and neurosyphilis and uninfected individuals. These results were directly compared to the rapid plasma regain (RPR) test, and the microhemagglutination assay for T pallidum (MHA-TP). P4-S3.08Results Screening results show high degrees of sensitivity and specificity for the Tp0326/Tp0453 chimeric construct, identifying all stages of syphilis infection from early primary to late latent. Conclusion The Tp0326/Tp0453 chimera shows promise as a new diagnostic antigen for detecting all stages of syphilis infection and for development into point-of-care diagnostic test formats. Objective Trichomonas vaginalis, a highly prevalent non-viral sexually transmitted infection, has been shown to be infected by a doublestranded RNA virus known as T vaginalis virus (TVV). The presence of this virus has been associated with increased trafficking of the immunogenic P270 to the surface of the parasite, and has therefore been hypothesised to be an important virulence factor in trichomoniasis. In the present study, we investigate the prevalence of TVV in globally distributed T vaginalis isolates and find an association between TVV infection and genetically distinct T vaginalis populations. Methods 150 T vaginalis isolates from the USA, Mexico, Italy, Southern Africa, Papua New Guinea and Australia were screened for TVV infection by running total RNA extract on 1% agarose gels to detect the presence or absence of the diagnostic 4.5 kb dsRNA genome of the virus. The prevalence of TVV in genetically distinct T vaginalis phylotypes was compared using c 2 tests. Results TVV was found to be present in 37% of T vaginalis isolates. We find a difference in the prevalence of TVV infection between genetically distinct populations of parasites, with 3% of phylotype 1 isolates containing the virus vs 73% of phylotype 2 parasites (<0.001). P4-S3.09 THE PREVALENCE OF TRICHOMONAS VAGINALISConclusions TVV prevalence varies between T vaginalis phylotypes 1 and 2. This finding has implications suggesting that TVV is transmitted vertically among parasites, as more closely relate...

208C, male samples were 100% reactive after 7 days storage, while female samples were 100% reactive after 6 days of storage. For samples stored at 308C, 100% reactivity was obtained at 2 days storage for male samples and 4 days storage for female samples. Conclusions This study shows urine samples from women infected with TV have a wide range of cell titres, with an average of w2000 TV cells/ml. TV can be detected for 14 days when stored refrigerated or for about 1 week at 208C. The use of sensitive, automated molecular tests such as the ATV assay for testing urine samples should facilitate screening for TV. Objective To observe the early inflammatory response by animal model with low genital tract Uu infection for investigating the relationship between the serovars and their pathogenicity. Methods Fifty 8e10-week-old, female BALB/c mice were randomly divided into five groups, namely, control group, serovar 1 group, serovar 3 group, serovar 4 group, and serovar 8 group, and then treated respectively with vaginal inoculation of liquid medium and Uu inoculae of serovars 1, 3, 4 and 8 after subcutaneous injection of oestradiol benzoate. The vulval symptoms and vaginal and cervical swabs were observed weekly after Uu inoculation. Four weeks later, all mice were sacrificed and their cervixes and vaginae were removed for histopathology. Results There was a significant difference in the inflammation severity of cervical mucosa (p<0.001e0.05) except vaginal mucosa among all groups. The inflammation severity of cervical mucosa in serovar 4 and 8 infection was superior to that in serovar 1 infection (p<0.01e0.05), but there was no statistical difference among other groups (p>0.05). Conclusions The results suggest that Uu could colonise and infect the cervical columnar epithelium other than vaginal squamous epithelium. Uu-induced inflammation response is different among its serovars. Serovars 4 and 8 are more virulent compared with biovar 1 such as serovar 1. Objective Trichomonas vaginalis, the causative agent of human trichomoniasis, is the most prevalent non-viral sexually transmitted infection and has been associated with increased risk of HIV transmission, making detection and treatment a global health priority. In this study, we evaluate the population genomics of globally distributed clinical isolates to characterise genetic diversity and identify population structure. Methods We use a panel of 21 microsatellite and three single copy gene markers to evaluate the population genomics of 18 clinical isolates collected from female patients attending New York City STD clinics in 2008, as well as 177 extant isolates collected from the USA, Mexico, Chile, Italy, Southern Africa, Papua New Guinea, and Australia. We use a panel of population genetic tools including Arlequin 3.11 and FSTAT to calculate expected heterozygosity (HE) and population differentiation (FST) statistics. To infer population structure, we use STRUCTURE 2.2, Network 4.516, and SeaView 4.2.4. We test for significance in clinical and demographic variables...