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PubMed1987

[Indirect immunofluorescence in the serodiagnosis of Trichomonas vaginalis infections].

By using immunofluorescent technique the Authors have observed that high titer circulating antibodies reacting with membrane or cytoplasmic structures of T. vaginalis occur in the majority (42/45) of women presenting because of trichomonas vaginitis. Only exceptionally high titer antibodies have been detected in serum of male subjects affected by urethral trichomoniasis. Low titer antibodies are found in serum of noninfected subjects and are supposed to be induced by cross reacting flagellate group antigens. The contribution of these findings to a better understanding of the natural history of trichomoniasis is briefly discussed.

Adult↗
PubMed1985

Trichomonas vaginalis infestation in sexually abused girls.

We report infection with Trichomonas vaginalis in four sexually abused girls; two of these patients are unique in that they were premenarchal. We discuss the clinical features in premenarchal and postmenarchal children and relate infection to the likelihood of prior sexual abuse.

Child↗
PubMed1996

Detection of Trichomonas vaginalis in endocervical and ectocervical smears.

The objective of this study was to determine the relative sensitivity of detecting Trichomonas vaginalis in endocervical as compared to ectocervical Papanicolaou-stained smears. The average number of organisms in 10 (200x) high-power fields (hpf) per smear was used for comparing the presence of trichomonads in the two smear types. In every pair of ectocervical-endocervical smears, there were always more trichomonads in the ectocervical smear. Fifty percent of cases showed at least 25 more trichomonads per 200x hpf in the ectocervical smear than in the endocervical smear. In 20 of 50 cases (40%), trichomonads were present only in the ectocervical smear. The organisms were found only in the endocervical smears of women with a heavier ectocervical trichomonad burden. In conclusion, trichomonads are detected in the Papanicolaou-stained ectocervical smear more often than in the endocervical smear; therefore, examination of an adequate ectocervical smear has greater utility than the endocervical smear for the diagnosis of trichomoniasis.

Animals↗
PubMed1998

Cytologic detection of Trichomonas esophagitis in a patient with acquired immunodeficiency syndrome.

Trichomonads are pathogens of the female genital tract and colonizers of the oropharynx. Gastrointestinal and pulmonary diseases have been reported in association with Trichomonas species, but a direct pathogenic effect of this organism in these organ systems remains controversial. Esophageal disease due to trichomonads has not been previously reported. A 43-yr-old man with acquired immunodeficiency syndrome (AIDS) with odynophagia and esophageal erosions was evaluated by endoscopy. Cytologic brushings from three of four sites in the esophagus were positive for trichomonads. Treatment with metronidazole resulted in clearance of the organism from the esophagus and improvement in clinical symptoms. We report esophageal trichomoniasis diagnosed on esophageal brush cytology in a man with AIDS. Clinical response was confirmed by cytologic studies and odynophagia improved with metronidazole treatment. Study of cytologic preparations was superior to biopsy for identification of this organism and was particularly useful in following the post-treatment course of disease.

Acquired Immunodeficiency Syndrome↗
PubMed2005

The diagnosis of trichomonas vaginalis in liquid-based Pap tests: morphological characteristics.

We investigated the morphological features of Trichomonas vaginalis in liquid-based Papanicolaou (Pap) (LBP) tests, in order to assess the impact of this new slide preparation system on recognition of T. vaginalis organisms. We reviewed 88 LBP test slides previously interpreted as showing T. vaginalis and assessed morphological characteristics of the organisms and the presence of secondary features. Eighty-six of 88 slides showed, at least focally, organisms with cytoplasmic granules, nuclei, and flagella allowing for confident recognition of T. vaginalis. Compared with 58 controls, cases of T. vaginalis were more likely to have squamous cells with perinuclear halos, "ghost cells," and coccobacilli. The lack of a "dirty" background or cytolysis allowed a more straightforward identification of the organisms. We concluded that T. vaginalis organisms are morphologically distinct in LBP tests, which may lead to an increased specificity when they are identified in such tests.

Adolescent↗
PubMed2005

The diagnosis of Trichomonas vaginalis in liquid-based Pap tests: correlation with PCR.

The conventional Papanicolaou smear (CPS) is not considered accurate for the diagnosis of Trichomonas vaginalis (T. vaginalis), and women noted to carry the organism on CPS are recommended to undergo confirmatory testing. Liquid-based preparations have been shown to facilitate the diagnosis of squamous lesions and may also facilitate the diagnosis of T. vaginalis. We used polymerase chain reaction (PCR) to investigate the accuracy of the diagnosis of T. vaginalis by the liquid-based Pap test (LBP). LBP with the diagnosis of T. vaginalis from a 12-mo period were identified. Residual samples from these cases were subjected to PCR for T. vaginalis as were the residual samples of a control group of 195 LBP (including 103 inflammatory LBP and 69 cases of atypical squamous cells) in which T. vaginalis was not diagnosed cytologically. PCR confirmed the presence of T. vaginalis in 50 of 51 (98%) LBP and identified 2 additional cases. Morphologic identification of T. vaginalis on LBP is highly accurate and should not require confirmatory testing.

Animals↗
PubMed1998

Trichomonas vaginalis: in vitro phagocytosis of lactobacilli, vaginal epithelial cells, leukocytes, and erythrocytes.

This paper explores the interaction of two strains of Trichomonas vaginalis, of high and low virulence, with the cell types present in the microenvironment of the parasite during human infections. With the use of transmission and scanning electron microscopy the sequence of internalization by T. vaginalis of Döderlein's lactobacilli, and of vaginal epithelial cells, leukocytes, and erythrocytes was documented. Furthermore, the degradation of ingested material by colocalization of acid phosphatase activity in phagocytic vacuoles was demonstrated. Phagocytosis of all cell types analyzed was found in both strains studied, although the highly virulent strain internalized target cells more rapidly than the less virulent one. Ultrastructural evidence indicated that phagocytosis takes place through two distinct mechanisms, only one involving the formation of a phagocytic stoma, characteristic of professional phagocytes. T. vaginalis phagocytosis may be both an efficient means of obtaining nutrients for the parasite and a significant factor in the pathogenesis of trichomonal infections of the human genitourinary tract.

Acid Phosphatase↗
PubMed1997

Extracellular release by Trichomonas vaginalis of a NADP+ dependent malic enzyme involved in pathogenicity.

This report presents evidence showing that Trichomonas vaginalis releases in the extracellular environment a functional form of NADP(+)-dependent malic enzyme. The protein which is likely responsible for the oxidative decarboxilase activity had already been identified in previous studies as P65, one of the five adhesive proteins of the protozoan. The same protein had also been described as AP65 by other authors, which identified it as one of the four surface proteins specifically responsible for binding of the parasite to the target cell in a ligand-receptor fashion. Gene characterization studies performed on P65 by different authors revealed that the nucleotide sequences of the genes coding for P65 display a striking homology with the ones coding for the trichomonad malic enzyme. The experiments performed in this work demonstrate that P65 is secreted and retains its adhesive properties in the extracellular environment, being able to bind both erythrocytes and HeLa cells. Therefore, an oxidative decarboxylase activity assay was performed on T. vaginalis cell-free filtrates, in order to assess if the released P65 displays cathalitic properties. The assay revealed that parasite-free supernatants exhibit an oxidative decarboxylase activity which is NADP(+)-dependent. On the basis of the most recent findings on T. vaginalis pathogenetic mechanism, which involves pH-dependent perforins, a role for the secreted enzyme as part of the system is proposed.

Animals↗
PubMed1999

The Trichomonas vaginalis phenotypically varying P270 immunogen is highly conserved except for numbers of repeated elements.

The prominent and phenotypically variable immunogenic protein of Trichomonas vaginalis, termed P270, is present in all isolates. Most, if not all, patients make antibody to the DREGRD epitope contained in the 333 bp tandemly repeating element (TRE). The complete sequence of p270 of a fresh clinical isolate was recently derived (Musatovova and Alderete, Microb Pathogen 1998; 24: 223-39). We hypothesized that the size polymorphisms of P270 were due to the varied number of TREs that comprise a large, central portion of the gene. In this study, we analysed the p270 coding regions of ten representative isolates. It was determined also that the sequence of the TRE of different p270 genes shared > or =99% identity, and individual TREs of the same p270 gene showed them to have identical nucleotide sequences, affirming the highly-conserved nature of this element within each gene. The coding regions upstream and downstream of the central TREs were then generated by PCR amplification using specific primers. The PCR products corresponding to the 5' and 3'-end coding, non-repeat sequences were then subjected to restriction analyses, and the regions were highly conserved for all p270 genes. The complete sequence of two p270 genes showed > or = 99% identity of amino acids at the N- and C-terminal regions of p270, further reinforcing that the reported polymorphisms in Mr of P270 is due to the varying number of TREs and, therefore, the size of the TRE domain. In support of this hypothesis and during these analyses, one isolate, T. vaginalis T016, was discovered which possessed a p270 gene with only one partial repeat unit. Importantly, and as with all other p270 genes, transcription of this single-repeat p270 gene in isolate T016 was confirmed. The start codon for the p270 T016 gene was preceded by the 12 nucleotide consensus Inr promoter-like sequence (TCATTTTTAATA) and possessed a putative transmembrane domain at the carboxy terminus.

Amino Acid Sequence↗
PubMed2000

A novel cysteine proteinase (CP65) of Trichomonas vaginalis involved in cytotoxicity.

The goal of this study was to demonstrate the participation in cellular damage of a Trichomonas vaginalis proteinase with a molecular mass of 65 kDa (CP65). By two dimensional gelatin-gel electrophoresis of trichomonad proteins we detected four spots with proteolytic activity on the 65 kDa region, but only one, pI 7.2, binds to the HeLa cell surface. By indirect immunofluorescence, rabbit antibodies against this proteinase localized the CP65 on the plasma membrane and in the cytoplasm of T. vaginalis. Pretreatment of parasites with the specific anti-CP65 antibody reduced trichomonal cytotoxicity to HeLa cell monolayers. The specific cysteine proteinase inhibitor, L-3-carboxy-2, 3-trans-epoxypropionyl-leucylamido (4-guanidino) butane (E64) abrogated the proteinase activity and reduced cytotoxicity levels of T. vaginalis in cell culture monolayers, indicating that the trichomonad CP65 is a cysteine proteinase. Activity of the CP65 proteinase was optimal at pH 5.5 and 37 degrees C, conditions similar to those of patients with trichomonosis. Also, this proteinase degraded some of the proteins found in the vagina, i.e. collagen IV and fibronectin, but not laminin-1 or haemoglobin. Finally, immunoprecipitation assays showed that sera and vaginal washes from trichomonosis patient possess anti-CP65 antibodies. In conclusion, results presented in this work demonstrate that the CP65 is a surface cysteine proteinase involved in T. vaginalis cytotoxicity to HeLa cell monolayers, as a virulence factor. It is immunogenic during human infection and degrades some extracellular matrix proteins, i.e. collagen IV and fibronectin.

Animals↗
PubMed2001

Binding of fibronectin by Trichomonas vaginalis is influenced by iron and calcium.

We have reported that Trichomonas vaginalis, the causative agent of the most common, non-viral sexually transmitted disease, bound to cover slips coated with fibronectin (FN) (Crouch & Alderete, Microbiol 1999 145: 2835-43). In this study, we extend that observation by showing that FN binding is specific, and we present data on the requirements of FN binding by T. vaginalis. Immunofluorescence and immuno-gold labelling readily detected FN throughout the trichomonal surface. Parasites bound to(125)I-labelled FN in a time- and concentration-dependent fashion. In the absence of protease inhibitor, iodinated FN was released from the trichomonad surface. Unlabelled FN specifically competed for binding in a concentration-dependent fashion with the(125)I-labelled FN. Interestingly, the amount of FN bound by T. vaginalis organisms was dependent on iron. High-iron-grown trichomonads acquired lower numbers of molecules but with 10-fold higher affinity than low-iron-grown organisms. Further, we show that for iron-replete organisms, calcium (Ca(2+)) at physiological levels increased amounts of bound FN. The increase in binding was rapid, occurring within 5 min of Ca(2+)addition, and required de novo protein synthesis. Co-incubation of live parasites with Ca(2+)in the presence of FN was necessary to increase the amount of FN bound. Treatment of trichomonads with okadaic acid, but not other phosphatase inhibitors, resulted in a 50% decrease in binding of FN, regardless of the presence of Ca(2+), suggesting a role for phosphatase in FN association. These results indicate that depending on the iron status of T. vaginalis organisms in vivo, Ca(2+)may influence trichomonad recognition and binding to FN during host parasitism.

Animals↗
PubMed1995

An improved method for in vitro susceptibility testing of Trichomonas vaginalis.

A protocol is presented for in vitro susceptibility testing of Trichomonas vaginalis. A 100 ml culture of the microorganisms is prepared for inoculation into antibiotic dilutions and controls by centrifugation, washing with 10 mM HEPES (pH 6.2) plus 1.5 x 10(-1) M NaCl, a second centrifugation and a resuspension in the HEPES-saline buffer. Inclusion of the gelling agent carrageenan in the culture medium permits an ease of harvesting the trichomonads and a reproducible initial cell density of 1-4 x 10(4) cells per ml. Following inoculation, tubes with antibiotic dilutions and controls are incubated anaerobically at 35 degrees C for 48 h, which corresponds to late exponential phase. Inclusion of a negative control helps determine Minimum Lethal Concentration (MLC) values.

Animals↗
PubMed1994

Analysis of the extracellular proteases of Trichomonas vaginalis.

We have previously isolated two extracellular cysteine proteases (60 and 30 kDa, respectively) from the cell filtrate of an isolate of Trichomonas vaginalis. In this study the clinical presentation of 12 clinical isolates of T. vaginalis was correlated with their protease activity. All 12 isolates produced a 60-kDa protease as demonstrated by immunoblotting. However, only 5 of 12 isolates produced a 30-kDa protease in the extracellular filtrate. Protease activity did not differentiate between isolates obtained from symptomatic versus asymptomatic women. The 60-kDa protease, which breaks down into a 43- and 23-kDa subunit, was detected by immunoblotting with anti-23-kDa cross-reacting rabbit serum in the vaginal washes of 3/3 women with active T. vaginalis infection, 0/1 woman cured of T. vaginalis and 0/2 control women with vaginal candidiasis. These results suggest that the 60-kDa protease is found in all isolates testes in vitro, is present in active disease and may be important in the pathogenesis of T. vaginalis infection.

Animals↗
PubMed1987

Trichomonas species in a subhepatic abscess.

A rare case of Trichomonas tenax/Trichomonas hominis and mixed oral bacterial flora in pus from a subhepatic abscess in a patient with a perforated penetrating ventricular ulcer is reported and the possible pathogenicity of the flagellate is discussed.

Humans↗
PubMed1986

[Transmission of Trichomonas vaginalis in swimming pools?].

Trichomonas vaginalis could not be isolated from the water of a highly frequented swimming pool of the City of Zurich. The protozoon T. vaginalis may show a slight degree of motility; even after 20 minutes exposure to the chlorinated water of a swimming pool; but it looses its infectivity within seconds after exposure to water of the swimming pool. It is not yet known how trichomonads are able to survive in the secretions of the vagina and on contaminated objects (fomites) in dressing rooms.

Female↗
PubMed1986

Further studies on the surface charge of various strains of Trichomonas vaginalis and Tritrichomonas foetus.

The surface charge of three strains of Trichomonas vaginalis and five strains of Tritrichomonas foetus was determined by direct measurement of the mean cellular electrophoretic mobility (EPM) of cells suspended in solutions of different ionic strength and pH. No differences were observed in the mean EPM among the two species, although significant differences among the strains exist. Strains that are more pathogenic to mouse, as measured using the subcutaneous assay, had a surface more negative. Treatment of the parasites with trypsin or neuraminidase reduced significantly their mean EPM and increased their isoelectric point. Tritrichomonas foetus was more sensitive to the enzyme treatment than T. vaginalis. Enzyme-treated cells recovered their normal EPM if, after enzyme treatment, they were incubated in fresh culture medium. The recovery process of trypsin-treated cells was inhibited 10-20% by addition of inhibitors of either protein synthesis (puromycin) or N-glycosylation of proteins (tunicamycin) to the incubation medium, suggesting that a cytoplasmic pool of sialoglycoproteins may exist. The recovering of the EPM of T. foetus and T. vaginalis previously treated with neuraminidase was inhibited by puromycin or tunicamycin about 40-50% and 17-30%, respectively. These observations suggest that sialoglycolipids exist on the surface of both parasite species, and that they contribute more to the surface charge of T. vaginalis than to that of T. foetus.

Animals↗
PubMed2003

Immunoblot analysis of Trichomonas vaginalis antigens recognized by rabbit hyperimmune serum raised against exoantignes.

The SDS-PAGE and immunoblot were used to identify Trichomonas vaginalis antigenic epitopes present in purified somatic and exo-antigens. The presence of common immunogenic proteins corresponding to molecular weights of 76, 60 and 23 kDa was revealed. Distinctive immunogenic bands of 92, 72, 55 and 40 kDa for the exo-antigens, and of 110, 80, 78 and 50 kDa for the somatic antigens, appeared when the antigens were probed by the homologous immune rabbit serum.

Adult↗
PubMed2006

Two novel serum-free media for the culture of Trichomonas vaginalis.

Definitive diagnosis of Trichomonas vaginalis, one of the common causes of sexually transmitted diseases in Turkey, relies on the microscopic examination of both fresh preparations and culture material of genital secretions and urine. The aim of this study was to compare the diagnostic efficacies of two culture media, serum-free TB1 and TB2, including iron and vitamin B12, respectively, with the well-known medium, TYM. Growth rate reached peak levels 48 h in TYM and 72 h in both TB1 and TB2 after inoculation. The highest amount of viable trophozoites has been obtained from TB1, almost equal to TYM but significantly higher than TB2. Fresh preparations obtained from the vaginal secretions of 119 patients have been examined and vaginal samples have been inoculated in TB1, TB2, and TYM. Viable T. vaginalis trophozoites have been detected in ten (8.4%) of fresh preparations and 11 (9.9%) of each medium. It is concluded that serum-free TB1 medium could be used effectively in both the isolation and maintenance of T. vaginalis culture in vitro.

Animals↗