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PubMed1984

Interaction between Trichomonas vaginalis and other pathogenic micro-organisms of the human genital tract.

Trichomonas vaginalis organisms were mixed with suspensions of Neisseria gonorrhoeae, Mycoplasma hominis or Chlamydia trachomatis to allow ingestion of these micro-organisms by the trichomonads. Culture studies indicated that gonococci and mycoplasmas were ingested and that the number of intracellular viable organisms decreased rapidly, most gonococci being killed within six hours and all mycoplasmas within three hours. Electron microscopy revealed phagocytic uptake and destruction of these two micro-organisms within the trichomonads, gonococcal degradation being associated with lysosomal enzyme activity. There was no evidence from cultural or electron microscopy studies that C trachomatis organisms persisted in mixed culture with T vaginalis.

Chlamydia trachomatis↗

PubMed1986

Morphological studies of the cytotoxicity of Trichomonas vaginalis to normal human vaginal epithelial cells in vitro.

In vitro cultured monolayers of normal human vaginal epithelial cells were incubated with axenic cultures of Trichomonas vaginalis. Two strains freshly isolated from patients with trichomoniasis and one strain that had been maintained in axenic culture for several years were studied. Freshly isolated trichomonads showed amoeboid movements, adherence to epithelial cell surfaces, and were cytotoxic to epithelial cells in vitro. In contrast, the laboratory strain maintained for years in axenic culture did not adhere to the epithelial cell monolayer and was only cytotoxic at a concentration 100 times that of freshly isolated trichomonads. Electron microscopy of monolayers exposed to T vaginalis for 24 hours showed that all epithelial cells in intimate contact with trichomonads had more or less disintegrated, whereas in monolayers exposed for six hours some of the epithelial cells in contact with T vaginalis were normal. T vaginalis organisms with amoeboid morphology contained a dense network of microfilaments in the part of the trichomonad that was in contact with an epithelial cell. Occasionally a pseudopodium was projected into the cytoplasm of disintegrated epithelial cells.

Cells, Cultured↗

PubMed1987

Action of anticytoskeletal compounds on in vitro cytopathic effect, phagocytosis, and adhesiveness of Trichomonas vaginalis.

The cytopathic effects of Trichomonas vaginalis treated with inhibitory concentrations of anticytoskeletal compounds (mebendazole, griseofulvin, colchicine, taxol, and cytochalasin B) were studied in mouse CLID fibroblast cultures. The evaluation, at different times, of cell numbers and morphological alterations showed that cytopathic effect was considerably reduced when protozoa were pretreated with mebendazole and griseofulvin, whereas colchicine, taxol, and cytochalasin B had less effect. Furthermore, treatment with mebendazole, griseofulvin, and colchicine decreased adhesiveness of the protozoan, whereas treatment with cytochalasin B and colchicine completely inhibited its phagocytic activity. From these results it may be concluded that alterations induced in the trichomonal cytoskeleton may affect its adhesiveness and its in vitro cytopathic effect, but there is no direct correlation between protozoan phagocytosis and its in vitro pathogenic effect.

Alkaloids↗

PubMed1988

Does lactobacillus vaccine for trichomoniasis, Solco Trichovac, induce antibody reactive with Trichomonas vaginalis?

Solco Trichovac is a vaccine of Lactobacillus acidophilus developed for treating trichomoniasis. The efficacy of the vaccine is reportedly due to cross reacting antibody being produced in people immunised with the lactobacillus bacteria. Several techniques, including enzyme linked immunosorbent, indirect immunofluorescence, immunoblot, and radioimmunoprecipitation assays were used to assess the extent of antigenic cross reactivity between Lactobacillus acidophilus and Trichomonas vaginalis. Data show a lack of antigenic relatedness between the Solco Trichovac lactobacilli and several strains of T vaginalis. Furthermore, antiserum to L acidophilus failed to inhibit trichomonad cytadherence or host cell killing, as is suggested by the producers of the Solco Trichovac vaccine.

Animals↗

PubMed1988

Differential susceptibility of fresh Trichomonas vaginalis isolates to complement in menstrual blood and cervical mucus.

The ability of complement in human menstrual blood and cervical mucus to kill Trichomonas vaginalis was compared with that of complement in serum, and 95 fresh trichomonal isolates obtained from vaginal wash material were evaluated for susceptibility to complement immediately after isolation. Only serum and menstrual blood with haemolytic activity produced total cytolysis of T vaginalis. The cytolytic abilities of these fluids were totally inactivated by treatment with heat or edetic acid (EDTA), which confirms the role of complement in cytolysis. Most cervical mucus samples had no detectable trichomonal cytotoxic properties. The cytotoxic activity in the remaining samples was not due to complement, as it was heat stable. Fresh isolates of T vaginalis and subpopulations of fresh isolates differed in their susceptibility or resistance to complement mediated lysis in serum. Resistance to complement did not remain stable after trichomonads were grown in vitro.

Adolescent↗

PubMed1989

Acquisition and retention of viruses by Trichomonas vaginalis.

Recently described occurrence of virus-like particles (VLP) in some strains of Trichomonas vaginalis suggests the possibility that the pathogenic significance of this organism may be broadened by its potential for viral transmission. Inasmuch as neither the source nor the host range of the VLP are known, any hazard which they may present for man cannot be estimated. A model has been established for the study of acquisition of known human viruses by T vaginalis. Tissue cultures were infected with two reoviruses and a fresh isolate of genital herpes simplex virus (HSV). A squirrel monkey reovirus was also included in the study. T vaginalis was inoculated into the virus cultures three days later. The progress of virus acquisition by the trichomonads was monitored by transmission electron microscopy and by culture. Virus-containing cell fragments were engulfed by trichomonads and internalised in vacuoles. After digestion of cellular debris only virus particle aggregates were retained. Viable reoviruses were recovered from the trichomonads for nine days, and HSV for six days. The results suggest the possibility of transmission of at least some viruses by T vaginalis.

Animals↗

PubMed1990

Properties of Trichomonas vaginalis grown under chemostat controlled growth conditions.

Trichomonas vaginalis isolates NYH 286 and IR 78 were grown in continuous flow culture conditions in a complex trypticase-yeast extract-maltose medium supplemented with heat-inactivated horse serum. Parasites could be stably maintained in the chemostat at high densities ranging from 1 x 10(6) to 1 x 10(7) organisms ml-1. Growth densities, acid production, and profiles of total versus secreted trichomonad proteins were characterised at different rates of growth and pH. Growth rate influenced the extent of parasite production of acid and the shedding of proteins into the medium but had no effect on overall parasite density. Lowering the pH from 6.0 to 5.0 resulted both in a decrease of cell density and acid production. At pH 4.5 isolate IR 78 but not NYH 286 was capable of growth and multiplication, showing the ability of some isolates to survive at the vaginal pH of healthy individuals. At this lower pH, however, isolate NYH 286 but not IR 78 synthesised new proteins which were detectable in stained gels. Also, inoculation of the chemostat with isolate NYH 286 comprising a mixture of fluorescent (positive, pos) and non-fluorescent (negative, neg) trichomonads as defined by monoclonal antibody reactivity to a surface immunogen resulted in a change in the parasite population to an almost homogeneous neg phenotype. These neg phenotype organisms switched back to pos phenotype after transfer to test tubes.

Animals↗

PubMed1997

The vagina has reducing environment sufficient for activation of Trichomonas vaginalis cysteine proteinases.

BACKGROUND: Trichomonas vaginalis, a worldwide distributed sexually transmitted protozoan, is remarkable for synthesis of numerous, distinct cysteine proteinases, the significance of which is evidenced by the presence in vivo of soluble proteinases in secretions and antiproteinase antibody in serum of patients with trichomonosis. These proteinases purportedly play a role in host parasitism and immune evasion. OBJECTIVE: It is known that for cysteine proteinases to be functional, they must be activated by disulphide reducing reagents. Whether or not the host vaginal environment has the reducing environment essential for activation of the trichomonad cysteine proteinases is unknown. Our goal, therefore, was to determine whether or not vaginal secretions had sufficient reducing power to activate the trichomonad proteinases. METHODS: 48 vaginal washes (VWs) from patients were assayed for reducing equivalents and a score in dithiothreitol (DTT) reducing equivalents was assigned to each VW. Activation of trichomonad cysteine proteinases was then tested under the range of reducing equivalents detected from VWs. The possible protective effect of hydrogen peroxide, an oxidising agent produced by some Lactobacillus species, on proteinase activity was also determined. RESULTS: Nine of 48 VWs (18.7%) possessed < or = 10 microM DTT reducing equivalents, four VWs (8.3%) had from 20 microM DTT to 40 microM DTT reducing equivalents, and most (50%) were between 10 microM to 15 microM. Overall, the range in VWs was from approximately 10 microM to 40 microM reducing equivalents. Importantly, data suggest differential proteinase activation over this in vivo range of reducing level. Only two T vaginalis cysteine proteinase activities were stimulated at 2.5 microM DTT in contrast with all proteinase activities present at 40 microM DTT, albeit quantitatively diminished compared with the activity at 1 mM DTT, the concentration routinely used in vitro. Finally, hydrogen peroxide reversibly neutralised all trichomonad proteinases. CONCLUSIONS: These results show that the vagina of women has a reducing environment adequate for activation of trichomonad proteinases. The data underscore that the host environment plays a role in the host-parasite interrelation. Finally, hypotheses can now be formulated to help explain resistance and susceptibility to infection commonly reported among women and between men and women with trichomonosis.

Animals↗

PubMed1997

A comparison of the sensitivity of the InPouch TV, Diamond's and Trichosel media for detection of Trichomonas vaginalis.

OBJECTIVE: This study compared the ability of three culture media (InPouch TV, Diamond's, and Trichosel) to support the growth of clinical isolates of Trichomonas vaginalis and their relative sensitivity for detection of the organism. METHODS: The majority of the clinical isolates were obtained from two San Francisco Bay Area clinics. T vaginalis was subcultured in 4 ml of one of the InPouch, Diamond's, or Trichosel media for 24-48 hours before evaluation. Twenty isolates were initially cultured in the InPouch test, 13 with Diamond's, and 10 with Trichosel. A haemocytometer was used to measure the initial concentrations of the organisms. Then serial dilutions were made in saline to yield approximately 2.0 x 10(4), 2.0 x 10(3), and 2.0 x 10(2) motile T vaginalis per ml. A 30 microliter inoculum from each dilution was transferred into 4 ml aliquots of the three media (387 individual tests, 43 x 3 dilutions x 3 media). Microscopic examinations for viable trichomonads were made at 24, 48, and 96 hours. Microscopy was through the pouch wall for the InPouch medium, and through a cover slipped slide with one drop of Diamond's and Trichosel media. RESULTS: At 24 hours, the InPouch demonstrated 84/129 positive, Diamond's 23/129, and Trichosel 18/129. At 48 hours, an accumulative positive rate for the InPouch was 98/129, for Diamond's 55/129, and Trichosel 47/129. At 96 hours the total positives for each test were 112/129 for the InPouch, 78/129 for Diamond's, and 74/129 for Trichosel. CONCLUSIONS: The InPouch TV test was significantly more sensitive than either Diamond's or Trichosel (at 0.01 level of significance, pInPouch > pDiamond's; pInPouch > pTrichosel on all three dilutions at 24, 48, and 96 hours). This increased sensitivity was the result of either a reduced generation time or the larger volume of media examined microscopically.

Animals↗

PubMed1989

Characterization and purification of extracellular proteases of Trichomonas vaginalis.

Extracellular protease activity was detected in serum-free culture filtrates of Trichomonas vaginalis. The activity was demonstrated by hydrolysis of hide powder azure and possessed the characteristics of cysteine type proteases: inhibition by N-ethyl maleimide, Cu2+, antipain, N-tosyl-L-phenylalanine chloromethyl ketone, N-tosyl-L-lysine chloromethyl ketone, leupeptin, chymostatin, and iodoacetamide, and enhancement by cysteine, EDTA, and dithiothreitol. The activity was optimal at acid pH and the protease was also active on peptide nitroanilides with arginine derivatives. Purification of this activity by ethanol precipitation, ammonium sulfate fractionation, ion exchange chromatography, and gel filtration resulted in the isolation of two proteases estimated by sodium dodecyl sulfate - polyacrylamide gel electrophoresis to have molecular masses of 60 and 30 kilodaltons (kDa), respectively. The larger molecular mass protease broke down during purifications to two subunits of approximately 23 and 43 kDa, as determined by gel electrophoresis. Rabbit sera derived by immunization with the 23-kDa subunit cross-reacted by immunoblot with the 60- and 43-kDa subunits, but not with the 30-kDa protease. These soluble products of T. vaginalis growth could be important pathogenically in establishing T. vaginalis infection in the normally acid (pH less than or equal to 4.5) environment of the vagina.

Animals↗

PubMed1993

Trichomonas vaginalis infection in pre-pubertal girls.

This short paper discusses the implications of Trichomonas vaginalis infection of the genital tract in children with respect to suspected child abuse. Two case reports are described to illustrate the problems that can arise in the investigation of such cases T. vaginalis is usually transmitted sexually but the potential for non-sexual transmission does exist.

Adolescent↗

PubMed2004

Toxicogenetic evaluation of metronidazole in the treatment of women infected with Trichomonas vaginalis.

Samples of peripheral blood were collected once from non-smoking women who were healthy (controls) and twice (immediately before and immediately after 7 days of treatment with metronidazole at 500 mg/day) from non-smoking women infected with Trichomonas vaginalis. Lymphocyte cultures were prepared and used, in toxicogenetic studies, to determine the frequency of sister-chromatid exchange (SCE), the mitotic index (MI), and the replication index (RI) for each sample. MTZ treatment of the infected women led to an increase in the frequency of SCE (P <0.001), a decrease in the MI (P <0.003), and a modification in the kinetics of cell proliferation, with a decrease in the RI (P <0.0006). The differences seen between the results for the controls and those for the infected women, before and after MTZ treatment, may be attributed to the presence of the parasite, to the treatment itself, and/or to variation in the host's response to infection with T. vaginalis.

Adult↗

PubMed1996

In vitro activity of metronidazole alone and in combination with clotrimazole against clinical isolates of Trichomonas vaginalis.

Trichomonas vaginalis vaginitis is generally treated with oral metronidazole. Widespread use of this drug has led, however, to selection of resistant strains. Topical therapy seems appropriate whenever systemic high dosage schedule cannot be adopted in order to overcome resistance. This study was designed to assess the activity of metronidazole alone and in combination with clotrimazole, an imidazole derivative for topical use, against T. vaginalis. Tests were performed employing the antitrichomonas activity of a fixed ratio of metronidazole with clotrimazole (5:1) which has been recently suggested for topical therapy and the checkerboard technique. All tests were carried out under aerobic conditions to maximize T. vaginalis resistance traits. Minimum inhibitory concentrations (MICs) of metronidazole for the 12 strains studied were in the range reported in the literature (0.5-32 micrograms/ml). The interaction of metronidazole with clotrimazole as assessed by the checkerboard technique gave an indifferent outcome with all the strains assayed (FIC = 1-2). The fixed concentration of drugs, however, produced synergism (FIC = 0.5) in 5 of 12 isolates. Spontaneous resistant strains were not selected from T. vaginalis exposed to sub-lethal levels of the drugs or by culturing a large inoculum in the presence of 1, 2, 4 and 8 times the MICs of metronidazole alone or in combination with clotrimazole. These results confirm and extend previous reports highlighting the good in vitro efficacy of the association of metronidazole plus clotrimazole against T. vaginalis.

Animals↗

PubMed2005

Heterologous expression in Tritrichomonas foetus of functional Trichomonas vaginalis AP65 adhesin.

BACKGROUND: Trichomonosis, caused by Trichomonas vaginalis, is the number one, nonviral sexually transmitted infection that has adverse consequences for the health of women and children. The interaction of T. vaginalis with vaginal epithelial cells (VECs), a step preparatory to infection, is mediated in part by the prominent surface protein AP65. The bovine trichomonad, Tritrichomonas foetus, adheres poorly to human VECs. Thus, we established a transfection system for heterologous expression of the T. vaginalis AP65 in T. foetus, as an alternative approach to confirm adhesin function for this virulence factor. RESULTS: In this study, we show stable transfection and expression of the T. vaginalis ap65 gene in T. foetus from an episomal pBS-ap65-neo plasmid. Expression of the gene and protein was confirmed by RT-PCR and immunoblots, respectively. AP65 in transformed T. foetus bound to host cells. Specific mAbs revealed episomally-expressed AP65 targeted to the parasite surface and hydrogenosome organelles. Importantly, surface-expression of AP65 in T. foetus paralleled increased levels of adherence of transfected bovine trichomonads to human VECs. CONCLUSION: The T. vaginalis AP65 adhesin was stably expressed in T. foetus, and the data obtained using this heterologous system strongly supports the role of AP65 as a prominent adhesin for T. vaginalis. In addition, the heterologous expression in T. foetus of a T. vaginalis gene offers an important, new approach for confirming and characterizing virulence factors.

Animals↗

PubMed2026

Long-read transcriptomics corrects Trichomonas vaginalis intron annotations and refines transcript-end features.

BACKGROUND: Trichomonas vaginalis causes the most prevalent non-viral sexually transmitted infection worldwide. Despite its large genome (181.5 Mb; 36,310 predicted protein-coding genes in NYU_TvagG3_2), intron annotations remain limited and inconsistently validated. A recent short-read RNA-seq study reported 63 putative active introns, but short reads can misassign splice boundaries and cannot resolve complete transcript structures. METHODS: We integrated Oxford Nanopore direct RNA sequencing (DRS), ONT cDNA long-read sequencing, and Illumina RNA-seq to refine intron annotations, transcript-end features, and UTR boundaries in T. vaginalis. Candidate introns were validated by targeted PCR and Sanger sequencing, and representative splicing events were further assessed using public SRA datasets. RESULTS: Starting from 31 historically annotated introns, motif-guided long-read screening and orthogonal validation identified 17 additional validated introns, increasing the curated set to 48 confirmed introns. Among these 17 events, three were previously unrecognized in the current NYU_TvagG3_2 reference annotation. We also corrected five reported loci, including two false-positive introns, two splice-coordinate misannotations, and one gene-sequence error. DRS further supported transcript termination site mapping, UAAA polyadenylation-signal profiling relative to poly(A) addition sites, and single-molecule poly(A)-tail estimation. StringTie mixed-mode assemblies provided updated UTR boundaries for intron-bearing transcripts and transcripts without curated introns. CONCLUSIONS: This study provides a rigorously validated, long-read-refined resource of intron annotations, UTR boundaries, and UAAA-guided transcript-end features for T. vaginalis, together with a reproducible workflow for non-model protists. These refinements improve the current reference annotation and support future studies of functional genomics, parasite biology, pathogenesis, and diagnostic development.

Trichomonas vaginalis↗

PubMed1979

Structure, protein composition and birefringence of the costa: a motile flagellar root fibre in the flagellate Trichomonas.

The costa is a rod-shaped intracellular organelle lying in the cytoplasm immediately below the undulating membrane. In certain large species of Trichomonas (T. gigantea, T. termopsidis and an undescribed species from the termite Porotermes adamsoni) this organelle is motile. Bending waves are transmitted along the length of the costa, in either direction. It is shown that the bending is actively produced by the costa itself. The form of the movements is described in detail. The costa is birefringent. At the point at which bending occurs there is a sharply localized reduction in birefringence. Electron microscopy shows that the costa is composed of longitudinally running lamellae, 2--3 nm thick and spaced 12 nm apart. These are connected to transversely running elements spaced at intervals of about 37 nm. The lamellae occur in two alternative configurations: straight and zig-zag. Bending probably results from a local, transient change from the straight to the zig-zag configuration. This would account for the local change in birefringence which accompanies bending. Polyacrylamide gel electrophoresis of isolated costas shows them to contain a number of protein components, of which the principal one has a molecular weight of about 90,000. Preliminary cytochemical evidence is presented for ATPase activity in the costa. The costa is a type of motile system distinct from any hitherto described.

Adenosine Triphosphatases↗

PubMed1988

The interaction of Trichomonas vaginalis and Tritrichomonas foetus with epithelial cells in vitro.

The Madin-Darby canine kidney epithelial cell line (MDCK) was used as a model for trichomonad-host cell interaction. Two laboratory strains of the human parasite Trichomonas vaginalis and the cattle's parasite Tritrichomonas foetus or their supernatants from axenic cultures were allowed to interact with confluent epithelial cultures. The interaction process studied by transmission and scanning electron microscopy revealed that both parasites adhere to monolayers through flagella, cell body and particularly for T. foetus, through the posterior projection of the axostyle. A close contact region between the trichomonad's surface and MDCK cells was observed. A study of the involvement of trichomonad surface component in the interaction process indicated that cytochalasin B treated-parasites adhere much less to epithelial monolayers than untreated parasites. Colchicine treatment did not affect such adhesion. Treatment of the parasites with trypsin reduced the adhesion of trichomonads to monolayers but did not interfere with the cytopathic effect. In contrast, treatment of the parasites with neuraminidase did not interfere with their adhesion to epithelial cells and the monolayer destruction was further increased.

Animals↗

PubMed1988

Localization of acetylated alpha-tubulin in Tritrichomonas foetus and Trichomonas vaginalis.

We used monoclonal antibodies specific for acetylated and nonacetylated alpha-tubulin to detect and to localize microtubules containing acetylated alpha-tubulin (stable microtubules) in the pathogenic protozoa Tritrichomonas foetus and Trichomonas vaginalis. SDS-PAGE analysis showed that tubulin is a major protein of both parasites, being enriched in cytoskeletal preparations of whole cells extracted with Triton X-100. The monoclonal antibodies, which recognize all isoforms of alpha-tubulin (B-5-1-2) and only acetylated alpha-tubulin (6-11B-1), bind to the tubulin of T. foetus and T. vaginalis as seen by immunoblotting. Tubulin-containing structures were localized using immunofluorescence microscopy and transmission electron microscopy of the whole cytoskeleton previously incubated in the presence of the anti-tubulin antibodies and a second antibody-gold complex, and then processed using the negative staining or replica techniques. The results obtained indicate that, in addition to the flagellar microtubules, those which form the peltar-axostyle system represent stable microtubules containing acetylated alpha-tubulin.

Acetylation↗