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Molecular probe for identification of Trichomonas vaginalis DNA.

Trichomoniasis is one of the most widespread sexually transmitted diseases in the world. Diagnosis can be achieved by several methods, such as direct microscopic observation of vaginal discharge, cell culture, and immunological techniques. A 2.3-kb Trichomonas vaginalis DNA fragment present in strains from diverse geographic areas was cloned and used as a probe to detect T. vaginalis DNA in vaginal discharge by a dot blot hybridization technique. This probe was specific for T. vaginalis DNA. It recognized strains from two regions in Italy (Sardinia, Piemonte) and from Mozambique (Africa). In addition, our probe did not cross-react with bacterial (Escherichia coli, Enterococcus spp., group B streptococci, Gardnerella vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, and Lactobacillus spp.), viral (herpes simplex virus type 2), fungal (Candida albicans), protozoan (Entamoeba histolytica, Giardia lamblia, Plasmodium falciparum, Leishmania major, and Leishmania infantum), or human nucleic acids. The probe reacted with Pentatrichomonas hominis and Trichomonas foetus. The limit signal recognized by our probe corresponded to the DNA of 200 T. vaginalis isolates. The 2.3-kb probe was used in a clinical analysis of 98 samples. Of these, 20 samples were found to be positive both with the probe and by cell culture, and only 14 of these were positive by a standard wet mount method.

Animals↗

Improved detection by DNA amplification of Trichomonas vaginalis in males.

Trichomoniasis is a sexually transmitted infection that is highly prevalent worldwide and has been linked to preterm birth and human immunodeficiency virus acquisition. In females, trichomoniasis causes vaginitis, while in males, it is frequently asymptomatic but can be a cause of urethritis. Control efforts have been hampered by the lack of a sensitive diagnostic technique for this infection in males. Men attending a sexually transmitted disease (STD) clinic for a new complaint were screened for Trichomonas vaginalis by culture and by PCR analysis of urine and urethral-swab specimens. The prevalence of Trichomonas determined by culture was 5% (15 of 300 specimens), compared to 17% (52 of 300) determined by PCR. Urine specimens yielded a greater number of positive results by PCR than did urethral-swab specimens. The sensitivity of PCR analysis of urine specimens in comparison to that of culture was 100%. The use of PCR techniques in urine specimen-based detection of T. vaginalis was highly sensitive and revealed a prevalence of infection more than three times that revealed by culture for men at high risk for STDs.

Animals↗

Pathogenic Trichomonas vaginalis cytotoxicity to cell culture monolayers.

Exposure of monolayer cultures of human urogenital and vaginal (HeLa), human epithelial (HEp-2), normal baboon testicular (NBT), and monkey kidney (Vero) cells to live pathogenic Trichomonas vaginalis resulted in extensive disruption of monolayers. Trypan blue was taken up by all host cells released from cell monolayers, which indicated irreversible damage of these cell types by trichomonads. Time and dose related data on cytotoxicity kinetics were obtained using increasing ratios of parasites to cells. All cell types were most sensitive to trichomonads at a multiplicity of infection of one. Release of tritiated thymidine (3H-thymidine) of the deoxyribonucleic acid (DNA) of prelabelled host cells after incubation with T vaginalis corroborated that extensive cytotoxicity was caused by pathogenic trichomonads in man. Only living parasites were cytotoxic, and no trichomonal toxic products were implicated in disruption of the cell monolayer cultures. A pathogenic bovine trichomonad, Tritrichomonas foetus KV-1, produced half as much cell damage as did T vaginalis. Trichomonas tenax, a non-pathogenic member of the normal flora of the oral cavity in man, produced no measurable cytotoxicity to HeLa cells when compared with the pathogenic human trichomonads.

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Vaginal antibody of patients with trichomoniasis is to a prominent surface immunogen of Trichomonas vaginalis.

Twenty vaginal washes (VWs) and ten vaginal mucus (VM) samples from patients with trichomoniasis were examined for the presence of antibody to surface protein immunogens of Trichomonas vaginalis. Fourteen of 20 VWs (70%) and 8 of 10 VM (80%) had immunoglobulin G (IgG) antibody (Ab) that reacted in an immunoprecipitation (IP) assay with one iodinated Trichomonas vaginalis surface protein immunogen with a relative molecular mass of 230,000 daltons (230-kDa) (P230). No similar IP of any iodinated protein was observed when detergent extract was first depleted of P230 with monoclonal antibody (MAb), indicating a highly specific VW IgG response of patients to P230. VWs were also obtained from 10 patients from one to four weeks after treatment. These VWs had the same, or in one case a greater, level of IgG to P230. Under no circumstances was Ab to P230 or any other trichomonad protein detected in VWs or VM from normal, uninfected women. Flow cytofluorometry with VW Ab yielded heterogeneous fluorescent and non-fluorescent populations of trichomonads, reaffirming the restricted Ab response to one or a few epitopes on P230 in the vagina of patients. Under identical conditions, the MAb gave totally fluorescent parasite populations of some isolates, and the MAb again demonstrated variable epitope accessibility to Ab binding (Infect Immun 1987;55:1037). Finally, the MAb or VW Ab was never cytolytic for immunoreactive (fluorescent) parasites, even in the presence of complement. This study identifies the most important trichomonad surface immunogen on the basis of the vaginal Ab response, and data underscore the significance of immune evasion strategies of this sexually transmitted disease agent.

Animals↗

Prevalence of Trichomonas vaginalis in a male sexually transmitted disease clinic population by interview, wet mount microscopy, and the InPouch TV test.

OBJECTIVE: To determine the prevalence of trichomoniasis in male patients from their urine at a Sexually Transmitted Disease (STD) Clinic using the InPouch TV culture system. METHODS: Two hundred and four patients were examined for STD infections. Their ages ranged between 17 and 72 years. Depending on their clinical symptoms tests were ordered for Neisseria gonorrhoeae, chlamydia, and for syphilis. Each patient submitted a clean catch urine specimen for trichomonas testing. A 15 ml aliquot of urine was centrifuged and a drop of the sediment examined microscopically. The remainder was cultured in the InPouch TV test. Each pouch was examined at 24 h, 48 h, and 5 days. RESULTS: Twenty-four of the 204 patients (12%) were culture positive for Trichomonas vaginalis and only three of these were wet mount positive. CONCLUSION: The InPouch TV test demonstrated an epidemiological important infected male population that was not indicated by wet mount microscopy.

Adolescent↗

The tampon test for trichomoniasis: a comparison between conventional methods and a polymerase chain reaction for Trichomonas vaginalis in women.

OBJECTIVES: Trichomonas vaginalis is the most common STD worldwide and the infection has been linked with an increased risk of HIV transmission. We present a detailed comparison between conventional collection and testing methods and the polymerase chain reaction (PCR) tampon test for T vaginalis. METHODS: Women were tested for the presence of T vaginalis by PCR analysis of a tampon specimen and by conventional methods which included one or more of the following: culture and microscopy from a high vaginal swab (HVS) or endocervical swab (ECS), and microscopy of a Papanicolaou (Pap) smear. RESULTS: T vaginalis was detected in 51/590 (8.6%) conventional tests and 93/590 (15.8%) tampon specimens. Retesting of all tampon PCR positive specimens confirmed 89/93 (95.7%) tests. Using the tampon PCR as the reference, the sensitivities of the different conventional sampling and testing methods for the detection of T vaginalis were 8.3% (5/60) for ECS microscopy or culture, 31% (13/42) for HVS microscopy or culture, 52.8% (19/36) for HVS directly inoculated into Trichomonas medium and 59.4% (38/64) for Pap smear. CONCLUSIONS: No conventional test in the remote setting has comparable sensitivity to PCR. The Pap smear is the next most sensitive, but requires a speculum examination. The use of PCR will allow inclusion of T vaginalis into STD screening programmes in both developed (lower prevalence) and developing (higher prevalence) countries.

Adolescent↗

Trichomonas vaginalis epidemiology: parameterising and analysing a model of treatment interventions.

BACKGROUND: Trichomonas vaginalis, which affects at least 170 million individuals globally, may increase the risk of transmission of HIV and predispose pregnant women to premature rupture of membranes and early labour. OBJECTIVE: To more clearly define the epidemiology of trichomoniasis and to develop a mathematical model of disease transmission dynamics in order to explore various treatment strategies. DESIGN: A deterministic model of trichomoniasis was constructed. Parameter values were set to fit the model to known endemic prevalence levels of Trichomonas vaginalis. Two treatment interventions ("screening" and "syndromic management") were simulated. RESULTS: The age specific prevalence of the disease was seen to differ from other STDs in a number of studies. Parameter fitting indicates that the average duration of infection in women is at least 3-5 years and approximately 4 months for men. "Syndromic management" (that is, treating only those with symptoms of disease) had minimal effect upon the endemic prevalence of disease even at high levels of coverage. "Screening" (that is, identification of individuals with both symptomatic and asymptomatic infection) was shown to be the most efficient method of control, but was sensitive to the screening interval. CONCLUSIONS: The control of trichomoniasis seems to have been a success in developed countries because of the regular access to health care, whereas it has remained endemic in many developing countries where control may only be possible by regular screening and treatment. However, without a large investment in services, success in controlling trichomoniasis is likely to be transitory.

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Concordance between genetic relatedness and phenotypic similarities of Trichomonas vaginalis strains.

BACKGROUND: Despite the medical importance of trichomoniasis, little is known about the genetic relatedness of Trichomonas vaginalis strains with similar biological characteristics. Furthermore, the distribution of endobionts such as mycoplasmas or Trichomonas vaginalis virus (TVV) in the T. vaginalis metapopulation is poorly characterised. RESULTS: We assayed the relationship between 20 strains of T. vaginalis from 8 countries using the Random Amplified Polymorphic DNA (RAPD) analysis with 27 random primers. The genealogical tree was constructed and its bootstrap values were computed using the program FreeTree. Using the permutation tail probability tests we found that the topology of the tree reflected both the pattern of resistance to metronidazole (the major anti-trichomonal drug) (p < 0.01) and the pattern of infection of strains by mycoplasmas (p < 0.05). However, the tree did not reflect pattern of virulence, geographic origin or infection by TVV. Despite low bootstrap support for many branches, the significant clustering of strains with similar drug susceptibility suggests that the tree approaches the true genealogy of strains. The clustering of mycoplasma positive strains may be an experimental artifact, caused by shared RAPD characters which are dependent on the presence of mycoplasma DNA. CONCLUSIONS: Our results confirmed both the suitability of the RAPD technique for genealogical studies in T. vaginalis and previous conclusions on the relatedness of metronidazol resistant strains. However, our studies indicate that testing analysed strains for the presence of endobionts and assessment of the robustness of tree topologies by bootstrap analysis seem to be obligatory steps in such analyses.

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Association of Trichomonas vaginalis with sociodemographic factors and other STDs among female inmates in Lisbon.

A study on STDs was conducted among 211 female inmates in a prison in Lisbon, Portugal, in order to establish possible associations between Trichomonas vaginalis infection, sociodemographic factors and other STDs. T. vaginalis was found in 31.2% of the women, from whom only 65.1% presented symptoms. It was more frequently isolated in the 20-40 year age group. On univariate analysis there was an association, although not statistically significant, between the existence of T. vaginalis, multiple sexual partners, drug addiction and no condom use. A statistically significant association was found between trichomoniasis, prostitution and other STDs. These were found in 87% of all women. T. vaginalis was detected alone in 23.8% (15/63) of all women with trichomoniasis, while 76.2% (48/63) of them had multiple infections with trichomonas and other STDs. A statistically significant association was present between trichomoniasis and Mycoplasma hominis and infection with Treponema pallidum. This study showed that the prevalence of T. vaginalis is as high as that of multiple infections with other STDs. Therefore, our findings seem to confirm that trichomoniasis serves as a marker for other STDs. Screening for STDs should then be offered to Portuguese inmates, or, at least, these women should be screened for T. vaginalis, as a marker for the other STDs.

Adult↗

Management of metronidazole-resistant Trichomonas vaginalis--a new approach.

Metronidazole-resistant trichomoniasis is an infrequent but challenging problem with no universally successful treatment. Since 1994, we have been using a combination regimen consisting of high-dose tinidazole plus a broad-spectrum antibiotic, i.e. doxycycline or ampicillin, and clotrimazole pessaries, for 7--14 days in this condition. A retrospective case review identified 11 cases of resistant trichomonas between 1994 and 2002. In the absence of resistance testing, resistance was defined clinically. Nine of the 11 patients were cured; one patient did not attend follow-up. In the women who attended for follow-up, the cure rate was 90%. We have found the combination of tinidazole, a broad-spectrum antibiotic, and clotrimazole pessaries to be a tolerable and effective treatment for metronidazole-resistant Trichomonas vaginalis. Although a variety of total doses of tinidazole were used in our patients, based on our findings, and those of others, we would recommend giving tinidazole 2 g twice daily for 14 days (total dose 56 g).

Ampicillin↗

Survey for Haemoproteus spp., Trichomonas gallinae, Chlamydophila psittaci, and Salmonella spp. in Galapagos Islands columbiformes.

Endemic free-ranging Galapagos doves (Zenaida galapagoensis) and introduced rock doves (Columba livia) were surveyed in several islands of the Galapagos archipelago to establish sample prevalence of hemoparasites, Trichomonas gallinae, Chlamydophila psittaci, and Salmonella species. A Haemoproteus sp., the only hemoparasite identified, was found in 89% of the Galapagos doves sampled but not in the rock doves. Trichomonas gallinae was detected by polymerase chain reaction in 44% of rock doves from San Cristobal but in none of the Galapagos doves. Chlamydophila psittaci was detected from cloacal swabs in 6% of the Galapagos doves but in none of the rock doves sampled. All positive cases of C. psittaci occurred on Española, where the crude sample prevalence was 24%. A polymerase chain reaction-based Salmonella test failed to show evidence of this organism from any birds sampled.

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Effect of virulent Trichomonas gallinae on the band-tailed pigeon.

Nine Trichomonas-free band-tailed pigeons (Columba fasciata fasciata) trapped in Colorado died from experimental infection with Jones' Barn Trichomonas gallinae 7.1 days (av.) post-inoculation. Three experimentally infected domestic pigeons (C. livia) used as controls died 5.7 days (av.) post-inoculation. Three band-tailed pigeons from Colorado naturally infected with avirulent T. gallinae. Trichomonads obtained from a fatal case of trichomoniasis in a band-tailed pigeon from California killed a band-tailed pigeon from Colorado; they did not kill five domestic pigeons but did induce severe oral caseation.

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A new method for the diagnosis of Trichomonas gallinae infection by culture.

Diagnosis of Trichomonas gallinae infection can reliably be made by the demonstration of organisms in material taken from a bird's mouth and crop. This can be examined directly or incubated in a growth medium until organisms are numerous enough to be easily found in aliquots examined with the microscope. We compared two methods of culture diagnosis of Trichomonas gallinae infection in pigeons. A commercially available kit, In-Pouch TF, proved as sensitive for this purpose as in vitro Diamond's medium, but had practical advantages over the usual in vitro system.

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Flagellar duplication and migration during the Trichomonas vaginalis cell cycle.

Trichomonas vaginalis is a flagellated protozoan, a representative of 1 of the earliest known eukaryotic lineages. Trichomonas vaginalis lacks centrioles but possesses basal bodies. We report here the cell cycle-dependent flagellar dynamics of T. vaginalis. By immunofluorescence, we found that T. vaginalis flagella duplicated during S-phase, segregated toward the nuclear poles, and then emanated from the spindle poles at mitosis. This behavior strongly parallels that of centrioles and other spindle pole-associated structures variously termed centrosomes, spindle pole bodies, or microtubule organizing centers. These observations support the hypothesis that flagellar forces may have provided motile forces for spindle pole alignment in an ancestral eukaryote.

Animals↗

[Pathogenic action and sero-immunologic phenomena of Trichomonas vaginalis].

Trichomonas vaginalis is a frequent parasite of the urogenital system of the human been. The pathological manifestations are different with each strain. These variations of the pathogenic action of the parasite are found in the study of the experimental pathogenic action of the animal and in vitro on cell culture. The sero-immunological phenomens which lead to the presence of the parasite are not completely known. Some technics was proposed for the diagnosis. Their practical utilisation is not completely related to the heterogenicity of the antigenic structure of the different strains of Trichomonas vaginalis.

Adult↗

Non-gonococcal urethritis (NGU) due to trichomonas vaginalis in Ibadan.

Two hundred and sixty two male patients attending the Special Treatment Clinic (STC) of the University College Hospital, Ibadan between July and December 1989, were examined for Trichomonas vaginalis as a cause of non-gonococcal urethritis using both microscopic and culture methods. Thirteen of the 262 male patients were the sexual partners of trichomonas vaginalis infected women. While 47(17.9) percent of the 262 men investigated had gonococcal urethritis, 215(82.1 percent) had non-gonococcal urethritis out of whom 18(8.4 percent) had trichomonal urethritis. The age range of peak incidence among the patients investigated is 20-29 years. A significant difference (P < 0.001) between the number of male sexual contacts that were positive for trichomoniasis (38 percent) and other male patients included in this study (8.4 percent) was demonstrated. Culture of the samples from the male patients investigated gave the highest proportion (100 percent) of positive results. This indicates the superiority of culture methods over other methods of diagnosing trichomoniasis such as microscopic method routinely used in our centre.

Adolescent↗

Prevalence of trichomonas vaginalis in patients with vaginal discharge in Lagos, Nigeria.

Trichomonas vaginalis is the most prevalent, non-viral sexually transmitted infection worldwide and probably the most commonly treatable in Africa and worldwide. The purpose of this study was to determine the prevalence of T. vaginalis among women with vaginal discharge attending a gynaecologic clinic in Lagos and to identify the common characteristics of the vaginal discharge in T Vaginalis. High vaginal swabs were taken from consenting adults with vaginal discharge. The quantity, colour and odour of the discharge were noted. Amine (10% KOH) test and pH were determined. Wet-mount microscopy Giemsa staining and culture in Oxoid (R) Trichomonas Medium were performed on each swab. A total of 200 patients were examined. One hundred and forty-nine (74.5%) had T. vaginalis. There was no statistically significant association between age, marital status, parity, number of sexual partners and prevalence of T vaginalis. The colour of the discharge was white in 104 (69.8%), yellow in 30 (20.1), clear in 15 (10.1%). None was frothy or greenish. The discharge was heavy in 50 (33.6%) and malodorous in 51 (34.2%). The pH range was 4-7 and 42 (28.2%) normal pH of4. In 47 (31.5%) the amine test was negative. The prevalence of T vaginalis among women with vaginal discharge is high. Women complaining of vaginal discharge should be thoroughly screened for T. vaginalis using all available methods.

Adolescent↗

Interactions between Trichomonas vaginalis and vaginal flora in a mouse model.

To study the role of vaginal flora and pH in the pathogenesis of Trichomonas vaginalis, an intravaginal mouse model of infection was established. By employing this model, the vaginal flora and pH of mice could be monitored for changes caused by the parasite. As a baseline, the endemic vaginal flora of BALB/c mice was examined first and found to consist mainly of Staphylococcus aureus and Enterococcus species (32-76%). Lactobacilli and enteric bacilli were moderate (16-32%) in their frequency of isolation, and the prevalence of both anaerobic species and coagulase-negative staphylococci was low (4-16%). Vaginal pH was recorded at 6.5 +/- 0.3. Estrogenization, which was required for a sustained T. vaginalis infection, did not significantly alter vaginal flora; however, a slight rise in the number of bacterial species isolated per mouse and a drop in vaginal pH (6.2 +/- 0.5) were observed. Trichomonas vaginalis-infected mice did not appear to show significant changes in vaginal flora although vaginal pH was slightly increased. This mouse model could have applications in both immunologic and pathogenic studies of T. vaginalis and, with further modifications, aid in the study of protist-bacterial interactions.

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