Trichomonas Detection in Leucorrhea Based on VIBE Method.

Trichomonas examination is one of the important items in the leucorrhea routine detection. And it cannot be recognized by still images because of the unstable morphology and unfixed focal location caused by motion characteristic. We proposed an improved VIBE algorithm. 6 videos (totally 1414 frames) are collected for testing. In order to compare the effects of the algorithms, we segment each frame artificially as ground truth. Experiments show that percentage of correct classification (PCC) achieves 88%. The proposed improved method can effectively suppress the false detection caused by the formed components such as epithelial cells in the leucorrhea microscopic image and the missed detection caused by the background model update during the movement. At the same time, improvements can effectively suppress smear and ghost areas. The algorithm proposed in this paper can be integrated into the leucorrhea automatic detection system.

Zoonotic Trichomonas tenax and a new trichomonad species, Trichomonas brixi n. sp., from the oral cavities of dogs and cats.

Trichomonads are known to inhabit the oral cavities of various mammals, including dogs, cats and horses. However, little attention has been paid to species identification, prevalence and zoonotic potential of these parasites, although their hosts live in close proximity with humans. According to the original description, oral trichomonads in dogs and cats belong to the genus Tetratrichomonas. Interestingly, later investigations suggested that the oral cavities of dogs and cats could be infected with different species of the genus Trichomonas, including the human oral cavity parasite Trichomonas tenax. Thus, in this study we investigated the occurrence of oral trichomonads in 111 domestic dogs and 122 cats using cell cultivation methods, nested PCR analyses, and the sequencing of ITS1-5.8rRNA-ITS2 regions. We found that both dogs and cats harbour T. tenax, with prevalences of 8.1% and 4.1%, respectively. Considerably more dogs were infected with different species of the genus Trichomonas (30.6%), which we also identified in cats (6.6%). An analysis of the potential risk factors suggested that dogs of more than 3years old or with dental disease signs are more frequently infected with Trichomonas sp. than younger dogs or dogs without the disease signs, and that crossbreed dogs revealed increased rates of infection in comparison with purebred dogs. An analysis of the cat population suggested that Trichomonas sp. infection is lower in younger and crossbreed cats. Although the morphology of Trichomonas sp. is very similar to that of T. tenax, based on a phylogenetic analysis of ITS1-5.8rRNA-ITS2 regions and the ssrRNA genes, we consider Trichomonas sp. to represent a new trichomonad species, for which we propose the name Trichomonas brixi.

Validation of wet mount microscopy against Trichomonas culture among women of reproductive age group in Western province, Sri Lanka.

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.

Observation of trichomonads infection in a child with periodontitis by direct microscopy at the dental office.

The pathogenicity of Trichomonas species is well documented. Although their exact involvement in gum disease is not fully understood, recent studies suggest a correlation between these protozoa and periodontitis. This case report details the first chair-side observation in Europe of an oral trichomonad infection in a child with periodontitis, by direct microscopy. The dramatic recovery of the patient, observed following administration of an anti-parasitic treatment, confirms the necessity of further investigation in this field.

Glycogen accumulation and degradation by the trichomonads Trichomonas vaginalis and Trichomonas tenax.

Several species of trichomonad have been shown to accumulate significant quantities of glycogen during growth, suggesting an important role for this compound in cell physiology. We provide the first analysis of the changes in glycogen content and glycogen phosphorylase activity that occur during in vitro growth of two trichomonad species: Trichomonas vaginalis and Trichomonas tenax. Both species accumulated glycogen following inoculation into fresh medium and utilized this compound during logarithmic growth. Glycogen phosphorylase activity also varied during growth in a species-specific manner. The expression of phosphorylase genes in T. vaginalis remained constant during growth and thus transcriptional control did not explain the observed fluctuations in phosphorylase activity. After cloning, expression, and purification, two recombinant glycogen phosphorylases from T. vaginalis and one recombinant glycogen phosphorylase from T. tenax had robust activity and, in contrast to many other eukaryotic glycogen phosphorylases, did not appear to be regulated by reversible protein phosphorylation. Furthermore, allosteric regulation, if present, was not mediated by compounds known to impact the activity of better characterized phosphorylases.

Cell death in trichomonads: new insights.

Tritrichomonas foetus is an amitochondriate parasite that possesses hydrogenosomes, unusual anerobic energy-producing organelles. In these organisms the "mitochondrial cell death machinery" is supposed to be absent, and the mechanisms that lead to cell demise remain to be elucidated. The presence of a cell death program in trichomonads has already been reported, suggesting the existence of a caspase-like execution pathway in such organisms. Here we demonstrate the alterations provoked by the fungicide griseofulvin and raise the possibility that other cell death pathways may exist in T. foetus. Dramatic changes in trichomonads morphology are presented after griseofulvin treatment, such as intense plasma membrane and nuclear envelope blebbing, nucleus fragmentation, and an abnormal number of oversized vacuoles. One important finding was the exposition of phosphatidylserine (PS) in the outer leaflet of the plasma membrane in cells after drug treatment, and also the presence of a high amount of misshapen flagella and tubulin precipitates as vacuolar contents, suggesting an autophagic process of abnormal cellular elements. Interestingly, immunoreactivity for activated caspase-3 was not detected during griseofulvin treatment, a finding distinct from the observed when this cell was treated with H(2)O(2). The possibility of the existence of different pathways to cell death in trichomonads is discussed.

A divergent ADP/ATP carrier in the hydrogenosomes of Trichomonas gallinae argues for an independent origin of these organelles.

The evolution of mitochondrial ADP and ATP exchanging proteins (AACs) highlights a key event in the evolution of the eukaryotic cell, as ATP exporting carriers were indispensable in establishing the role of mitochondria as ATP-generating cellular organelles. Hydrogenosomes, i.e. ATP- and hydrogen-generating organelles of certain anaerobic unicellular eukaryotes, are believed to have evolved from the same ancestral endosymbiont that gave rise to present day mitochondria. Notably, the hydrogenosomes of the parasitic anaerobic flagellate Trichomonas seemed to be deficient in mitochondrial-type AACs. Instead, HMP 31, a different member of the mitochondrial carrier family (MCF) with a hitherto unknown function, is abundant in the hydrogenosomal membranes of Trichomonas vaginalis. Here we show that the homologous HMP 31 of closely related Trichomonas gallinae specifically transports ADP and ATP with high efficiency, as do genuine mitochondrial AACs. However, phylogenetic analysis and its resistance against bongkrekic acid (BKA, an efficient inhibitor of mitochondrial-type AACs) identify HMP 31 as a member of the mitochondrial carrier family that is distinct from all mitochondrial and hydrogenosomal AACs studied so far. Thus, our data support the hypothesis that the various hydrogenosomes evolved repeatedly and independently.

Cytokinesis arrest and nuclear fission in low density populations of trichomonad protozoan.

Cell growth of anaerobic protozoan Tritrichomonas foetus was analyzed. This protozoan usually proliferates in extremely high density, but protozoan parasites were dispersed uniformly in F-bouillon medium and cell division stopped temporarily. However, nuclear fission continued and giant polynucleated cells formed. Later, cell division resumed and cells returned to normal form. In conditioned medium, cytokinesis of the dispersed parasites did not stop. Results indicated that T. foetus cells secreted an extracellular factor that influenced cytokinesis.

Special organelles of some pathogenic protozoa.

Parasitic protozoa comprise a large number of species, some of which are agents of important diseases. They are also of interest from the point of view of cell biology since they contain special organelles and structures. This review analyses our present knowledge of (1). the glycosomes, found in members of the Kinetoplastida order, (2). the hydrogenosomes found in some anaerobic protozoa, especially in trichomonads, (3). the acidocalcisomes, recently described in several protozoa, and (4). structures and organelles participating in the endocytic pathway in trypanosomatids.

Tritrichomonas foetus: induced division synchrony by hydroxyurea.

Treatment of cultures of Tritrichomonas foetus with 4 mM hydroxyurea (HU), a known DNA synthesis inhibitor, induced pseudocyst formation and caused a mitotic burst. An hour after drug release there was a characteristic, synchronous burst of cell division. T. foetus culture was arrested in the G2/M phase. The synchrony index varied from 66% to 69%. The synchrony was maintained for several cell cycles, even in thawed cultures which had been frozen for storage in liquid nitrogen. The synchronized cells were analyzed by light and scanning electron microscopy, as well by flow cytometry.

Morphologic aspects of Tetratrichomonas didelphidis isolated from opossums Didelphis marsupialis and Lutreolina crassicaudata.

Tetratrichomonas didelphidis (Hegner & Ratcliffe, 1927) Andersen & Reilly, 1965 is a flagellate protozoan found in the intestine, cecum, and colon of Didelphis marsupialis. The parasitic protozoa used in this study was found and isolated in the intestine of opossums in Pavlova starch-containing medium in Florianópolis, State of Santa Catarina, Brazil, from D. marsupialis and Lutreolina crassicaudata. The strains were cultivated in Diamond medium without maltose and with starch solution, pH 7.5 at 28 degrees C. The specimens were stained by the Giemsa method and Heidenhain's iron hematoxylin. The light microscopy study of the trophozoites revealed the same morphologic characteristics as specimens previously described.

Strategies by which some pathogenic trichomonads integrate diverse signals in the decision-making process.

The interaction between each one of Trichomonas vaginalis and Tritrichomonas foetus with their hosts is a complex process in which components associated to the cell surfaces of both parasites and host epithelial cells, and also to soluble components found in vaginal/urethral secretions, are involved. Either cytoadhesion or the cytotoxicity exerted by parasites to host cells can be dictated by virulence factors such as adhesins, cysteine proteinases, laminin-binding proteins, integrins, integrin-like molecules, a cell detachment factor, a pore-forming protein, and glycosidases among others. How trichomonads manipulate informations from the extracellular medium, transduce such informations, and respond to them by stimulating the activities of some surface molecules and/or releasing enzymes are the aspects concerning trichomonal virulence which are here briefly reviewed and discussed.

Strategies by which some pathogenic trichomonads integrate diverse signals in the decision-marking process

The interaction between each one of Trichomonas vaginalis and Tritrichomonas foetus with their hosts is a complex process in which components associated to the cell surfaces of both parasites and host epithelial cells, and also to soluble components found in vaginal/urethral secretions, are involved. Either cytoadhesion or the cytotoxicity exerted by parasites to host cells can be dictated by virulence factors such as adhesins, cysteine proteinases, laminin-binding proteins, integrins, integrin-like molecules, a cell detachment factor, a pore-forming protein, and glycosidases among others. How trichomonads manipulate informations from the extracellular medium, transduce such informations, and respond to them by stimulating the activities of some surface molecules and/or releasing enzymes are the aspects concerning trichomonal virulence which are here briefly reviewed and discussed.

Similes and metaphors I: A nature walk through benign gynecologic cytology.

Similes and metaphors useful in benign gynecologic cytology are presented. Included are those pertaining to normal cellular elements, organisms, contaminants, and benign changes.

[Morphology and diagnosis of Entamoeba gingivalis and Trichomonas tenax and their occurrence in children and adolescents]. / Morfológia a diagnostika Entamoeba gingivalis a Trichomonas tenax a ich výskyt u detí a mládeze.

Specimens obtained from 176 eight- to nineteen-year-old subjects were cultured for oral protozoa. Of the given series, 25 subjects (14.2%) had in their mouth only E. gingivalis, 5 (2.9%) had a mixed invasion of E. gingivalis and T. tenax, and 2 subjects (1.1%) had only T. tenax. A total of 32 subjects (18.2%) were infested by oral protozoa. The following conclusions were made: oral protozoa occur also in children and teenagers with a cured or intact dentition; their occurrence rate was higher in 11- to 19-year-old subjects than in the lower age groups; both protozoa can occur simultaneously; their occurrence rate was age dependent (increasing with age) with the rate of E. gingivalis rising significantly more rapidly with age than that of T. tenax; their occurrence was found to be sex dependent with higher rates in boys than in girls. In the light of the high recovery rate (83.3%) of E. gingivalis from dental and periodontal swabs, not only specimens of saliva should be collected but also dental nad periodontal swabs should be taken and cultured when studying the occurrence of oral protozoa in the population. (Tab.4,Fig.2,Ref.7).

Cell-associated and extracellular proteolytic activity of an oral flagellate, Trichomonas tenax.

Proteolytic activities in crude extracts and culture filtrates from Trichomonas tenax were determined using hide powder azure as substrate and the proteinase profiles in both samples were analysed in SDS-polyacrylamide gels containing copolymerized gelatin. The enzyme activity in the crude extract was detected over a broad pH range and was strongly activated by dithiothreitol, mainly in the pH range 5-8, and inhibited by cysteine proteinase inhibitors. Extracellular enzyme activity in culture filtrates was SH-dependent and increased continuously during incubation of the cell suspension, suggesting proteinase release. A total of seven distinct proteolytic bands could be detected in crude preparations. Three of these, with apparent Mr values 35,000, 45,000 and 56,000 and a pH optimum of 4-7, were SH-dependent and their inhibitory sensitivities were characteristic for cysteine proteinases. The 45,000 and 56,000 proteinases probably corresponded to those found in the culture filtrates. Proteolytic bands with apparent Mr 76,000, 87,000, 102,000 and 270,000 and pH optima in the alkaline region, pH 8-9, were independent of SH groups and were inhibited by a chelating agent EDTA, suggesting that they belong to the metalloproteinase family.

Pesquisa de Ureaplasma urealyticum pregnant women and the correlation with the presence of Trichomonas sp / Research of Ureaplasma urealyticum pregnant women and the correlation with the presence of Trichomonas sp

Foram feitas pesquisas de Ureaplasma urealyticum em material cervical e de Trichomonas sp no conteúdo vaginal de 74 gestantes. A existência ou näo de relaçäo entre a presença simultânea de Ureaplasma urealyticum e Trichomonas sp foi analisada, uma vez que os dados fornecidos pela literatura internacional säo bastante controvertidos. Trichomonas sp estava presente em 21,5% das amostras positivas para Urealyticum e em 13% das amostras em que a ausência do micoplasma foi constatada. Estatisticamente essas diferenças näo foram significantes impossibilitando inferir qualquer correlaçäo entre os dois achados

Pulmonary trichomoniasis and Trichomonas tenax.

Pulmonary trichomoniasis is usually caused by aspirated Trichomonas tenax. Adult men with chronic purulent or necrotic pulmonary disease are usually affected. Sixty-eight patients were previously described. A Russian study demonstrated pulmonary trichomoniasis in 19 of 112 patients (17%), mostly in patients with lung cancer, lung abscess, or bronchiectasis. Rarely, pulmonary trichomoniasis may be caused by an intra-abdominal (T. hominis) or genitourinary (T. vaginalis) infection. T. tenax is usually regarded as a harmless commensal of the human mouth. Its prevalence ranges from 4% to 53% and may exceed that of vaginal infection with T. vaginalis in adult females. It is frequently found in patients with poor oral hygiene. Cultural identification is superior to microscopic examination of wet-smear, gram-stained and Papanicolaou-stained preparations. Aspirated pulmonary trichomoniasis is an opportunistic infection. Until the question of possible pathogenicity is resolved, metronidazole should be given. The underlying pulmonary disease should be vigorously treated.