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PubMed2006

Trichomonas vaginalis detection using real-time TaqMan PCR.

1978 women and 93 men, all suspected of having a Trichomonas vaginalis infection, were tested for the presence of T. vaginalis by real-time PCR using the T. vaginalis-specific 2-kb repeated sequence, and by direct microscopy and culture. 40 samples were positive by T. vaginalis real-time PCR and 27 were positive by wet mount microscopy, either direct or after culture. All samples positive by direct microscopy of culture were also positive by real-time PCR. Of the 13 samples which were real-time PCR positive but negative by direct microscopy and culture 11 were confirmed by another T. vaginalis real-time PCR based on the beta tubulin gene. Only 2 samples (0.1%) showed inhibition in the PCR. The prevalence of T. vaginalis infection in the female patients was 1.8%. The sensitivity, specificity, positive and negative predictive values of the real-time PCR were 100%, 99.9%, 95% and 100%, respectively. The same test characteristics for the combined conventional T. vaginalis detection methods (microscopy+culture) were 71%, 100%, 100% and 99%, respectively. Therefore, real-time PCR is the method of choice for the diagnosis of T. vaginalis infection.

Animals↗
PubMed2004

Trichomonas vaginalis: random amplified polymorphic DNA analysis of isolates from symptomatic and asymptomatic women in India.

Trichomonas vaginalis, the causative agent for human trichomoniasis, is a protozoan parasite. Trichomoniasis is the most common non-viral sexually transmitted disease. The infection in women may be asymptomatic or may lead to severe vaginitis, cervicitis and severe sequelae. Despite its high prevalence, the genetic variability and factors leading to symptomatic infection have been poorly understood. One thousand women in childbearing age group were screened for the presence of T vaginalis. Thirty-eight women were found positive for T vaginalis and out of these 22 (57.9%) were having symptomatic infection and 16 (42%) were asymptomatic. Fresh isolates from 15 symptomatic and 15 asymptomatic women were axenised and subjected to random amplified polymorphic DNA (RAPD) analysis with the use of five different random primers (OPD 1-OPD 5). The isolates with similar banding pattern were assigned as a single type. OPD 3 indicated least (nine types) while OPD 4 indicated highest typing (18 types) ability. Phylogenetic analysis using RAPD distance software indicated two distinct lineages; upper branch consisting of only seven symptomatic isolates while lower branch consisting of all the 15 asymptomatic isolates the other eight symptomatic isolates were recorded in separate cluster. The study indicated that RAPD technique might be helpful to delineate the pathogenic mechanism(s) for its virulence; however, further studies on large number of isolates are desired to elucidate the findings.

Animals↗
PubMed2006

Characterization of an ecto-5'-nucleotidase (EC 3.1.3.5) activity in intact trophozoites of Trichomonas gallinae.

This study describes the enzymatic properties of an ecto-5'-nucleotidase in Trichomonas gallinae. The enzyme hydrolyzes nucleoside monophosphates at pH 7.2 and is activated by divalent cations, such as magnesium. Ecto-5'-nucleotidase activity was insensitive to levamisole, tetramisole (alkaline phosphatase inhibitors), and AMPCP (adenosine 5'-[alpha,beta-methylene]diphosphate), an ecto-5'-nucleotidase inhibitor, whereas 0.1mM ammonium molybdate (considered a potent inhibitor of 5'-nucleotidase activity) completely inhibited the enzyme activity. The apparent K(M) (Michaelis constant) and Vmax (maximum velocity) values for Mg2+-AMP were 466+/-57 microM and 3.7+/-0.59 nmolPi/min/10(6) trichomonads, respectively. Considering that trichomonads lack the ability to synthesize purines and pyrimidines de novo, the presence of an ecto-5'-nucleotidase in intact trophozoites of T. gallinae could be important in regulating the extracellular nucleotide levels and generating adenosine, essential for the survival strategies of the parasite.

5'-Nucleotidase↗
PubMed1980

Three metronidazole-resistant strains of Trichomonas vaginalis from the United States.

Trichomonas vaginalis isolates were obtained in the United States from three women with trichomoniasis refractory to routine treatment with metronidazole. Cure was obtained in these cases with increased dosage of metronidazole. In experimental mouse infections, all three strains showed a significantly (about 10 to 20 times) lower susceptibility to metronidazole than did a number of control strains. In vitro, the lowered susceptibility could be detected only in an aerobic assay but not in the routine anaerobic assay. The conclusion is that T. vaginalis strains with lowered metronidazole susceptibility do exist. Such strains might be responsible for a certain, but as yet entirely unknown, number of failures in routine metronidazole therapy of human trichomoniasis.

Adult↗
PubMed1996

Detection of Chlamydia trachomatis and Trichomonas vaginalis by polymerase chain reaction in introital specimens from pregnant women.

OBJECTIVE: We analyzed whether specimens obtained from the vaginal introitus were comparable to endocervical and posterior vaginal vault specimens for detection of Chlamydia trachomatis and Trichomonas vaginalis by polymerase chain reaction. STUDY DESIGN: Introital and endocervical specimens were obtained from 300 women at the first prenatal visit. Specimens from the posterior vaginal vault were also obtained from 219 of these patients. All samples were tested for C. trachomatis and T. vaginalis by polymerase chain reaction. RESULTS: C. trachomatis was identified in the endocervices of 36 women (12.0%); all but one of these women and none of the endocervical-negative women were positive for this organism in the introitus. T. vaginalis was detected in 22 women (10.0%); all but one were also introitus positive for this organism. Compared with endocervical and vaginal polymerase chain reaction, introital testing had a 100% specificity and a 97.2% and 95.5% sensitivity for detecting C. trachomatis and T. vaginalis, respectively. CONCLUSION: Polymerase chain reaction analysis of vaginal introital specimens is highly sensitive and specific in detecting C. trachomatis and T. vaginalis in pregnant women and thus provides an alternative to speculum examination in screening for these pathogens.

Adult↗
PubMed1996

Evaluation of a deoxyribonucleic acid probe for the detection of Trichomonas vaginalis in vaginal secretions.

OBJECTIVE: Our purpose was to compare Affirm VP, a new deoxyribonucleic acid probe test, with standard "wet preparation" microscopic examinations and culture for the identification of Trichomonas vaginalis organisms in vaginal secretions. STUDY DESIGN: We examined vaginal samples from 615 women with symptoms or signs of vaginitis for T. vaginalis using the deoxyribonucleic acid probe test, microscopic examination of wet preparations of vaginal secretions, and culture in modified Diamond's medium. RESULTS: T. vaginalis was identified in specimens from 95 (15.4%) of the 615 patients. Cultures in Diamond's medium identified 93 (98%) of the 95 infected patients. Vaginal wet preparation identified 76 (80%) of the infected women. The deoxyribonucleic acid probe test detected 86 (90.5%) of the 95 infected patients. There was one false-positive deoxyribonucleic acid probe test (specificity 519/520: 99.8%). CONCLUSION: The Affirm VP deoxyribonucleic acid probe test had a sensitivity of 90% and a specificity of 99.8% for the identification of T. vaginalis organisms in women with symptoms with a high prevalence of trichomoniasis. Such a nonculture test may be of considerable benefit in diagnosing T. vaginalis infections, especially in settings where microscopy, culture, or both are unavailable, inconvenient, or unreliably performed.

Animals↗
PubMedDate not supplied

Trichomonas vaginalis: evaluating capsid proteins of dsRNA viruses and the dsRNA virus within patients attending a sexually transmitted disease clinic.

Some isolates of Trichomonas vaginalis, the number one, non-viral sexually transmitted disease agent, are infected with one or several distinct double stranded (ds)-RNA virus. Immune rabbit anti-capsid serum (IRS) reacted with the capsid protein of purified dsRNA virus of a subset of the virus-infected T. vaginalis isolates. A monoclonal antibody (mAb) that recognized the capsid protein reactive with the IRS was generated. Analysis of the virus capsid protein of virus-infected isolates by probing nitrocellulose blots with mAb revealed diversity among immunoreactivity and in the size of the reactive capsid protein. Despite difficulties in visualizing virus within parasites by cross-section electron microscopy, gold-conjugated mAb readily labeled the cytoplasm of virus-positive trichomonads. Finally and importantly, isolates infecting patients attending an STD clinic, 75% of which were virus-positive isolates, had capsid protein of the same size detected by mAb present in all dsRNA viruses.

Animals↗
PubMed2003

Pulmonary coinfection by Trichomonas vaginalis and Pneumocystis sp. as a novel manifestation of AIDS.

A 41-year-old man was hospitalized, presenting increasing dyspnea and extensive ground-glass opacities on chest X-ray. Infection by human immunodeficiency virus was confirmed. Cytologic examination of bronchoalveolar lavage fluid revealed numerous trichomonads and aggregates of Pneumocystis sp. Treatment was followed by rapid improvement of respiratory symptoms and chest X-ray. The trichomonad species found in the lungs was identified as Trichomonas vaginalis by small-subunit rRNA gene amplification and sequencing. With the exception of rare cases of contamination of newborn babies during delivery, T. vaginalis has never been found in lungs in healthy or immunocompromised adults. In the present case, T. vaginalis is found as coinfecting agent. Our data, like those found in the literature, suggest that trichomonads are overlooked parasites that may be regularly implicated in diverse human pathologies.

AIDS-Related Opportunistic Infections↗
PubMed2001

Host and pathogen interaction during vaginal infection by Trichomonas vaginalis and Mycoplasma hominis or Ureaplasma urealyticum.

Vaginal infections by Trichomonas vaginalis and Mycoplasma hominis have been shown to be associated. Since M. hominis and Ureaplasma urealyticum are similar pathogens, both belonging to the class of the mycoplasmata, we describe here a molecular study into the interdependence of U. urealyticum and T. vaginalis during infection. Susceptibility towards infection by U. urealyticum depends on genetic polymorphism in the interleukin-1 receptor antagonist (IL-1RA) gene. Now, we defined the relation between IL-1RA genotypes and infection by M. hominis and T. vaginalis. Finally, we also developed a restriction fragment length polymorphism (RFLP) tool for mapping variation in the T. vaginalis AP33 adhesin in order to define putative associations between parasite subtype and mycoplasmata or host. Studies using crudepellets from T. vaginalis culture broth clearly confirm the association between T. vaginalis and M. hominis infection. The association between IL-1RA genotype 2,2 and lack of U. urealyticum infection is corroborated as well. U. urealyticum infection and infection by T. vaginalis are independent. Furthermore, T. vaginalis and M. hominis infection are not depending on IL-1RA genotypes. Interestingly, one of the three AP33 RFLP types identified appeared to be associated with the absence of U. urealyticum infection. In conclusion, the complex interaction between bacterial and parasitic pathogens and the infected host is determined by genetic characteristics of host and microorganisms involved.

Adult↗
PubMed2002

Trichomonas gallinae: a possible contact-dependent mechanism in the hemolytic activity.

The in vitro hemolytic activity of Trichomonas gallinae was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of six adult animals of different species (rabbit, rat, chicken, horse, bovine, and sheep). Results showed that T. gallinae lysed all human erythrocytes groups, as well as rabbit, rat, chicken, horse, bovine and sheep erythrocytes. No hemolysin released by the parasites could be identified. Hemolysis did not occur with trichomonad culture supernatants, with sonicated extracts of T. gallinae, or with killed organisms. The scanning electron microscopy (SEM) showed that the erythrocytes adhered to the parasite surface and were phagocytosed. These observations suggest that the contact between T. gallinae and erythrocytes may be an important mechanism in the injury caused to the erythrocytes. The hemolytic activity of T. gallinae may be an efficient means of obtaining nutrients for the parasite and allow the investigation of the mechanism used by T. gallinae to damage cellular membranes.

Animals↗
PubMed1998

In vivo and in vitro sensitivity of Trichomonas gallinae to some nitroimidazole drugs.

A study was carried out in order to establish the in vivo efficacy of two nitroimidazole drugs against the flagellate Trichomonas gallinae. The results obtained showed a high proportion of therapeutic failures. Thus, carnidazole and dimetridazole failed to eliminate infection in 13 of 17 and 20 of 21 racing pigeons, respectively. The in vitro susceptibility of four T. gallinae isolates to five nitroimidazolic drugs, that is, carnidazole, dimetridazole, metronidazole, ornidazole and ronidazole, was also determined. Minimal lethal concentrations (MLCs) of these drugs were obtained. One of the isolates tested had already proved to be resistant in the first in vivo tests, while the other three had been obtained from wild birds. We were able to confirm in vitro the resistance of the racing pigeon isolate to all the nitroimidazole drugs tested. For carnidazole, dimetridazole, metronidazole and ornidazole the MLC ranged between 93.75-500 microg ml. Although ronidazole showed a greater potency than the other nitroimidazole derivatives against this isolate (MLC: 15.62-31.25 microg ml), 8-22 times more drug was necessary for efficacy against this isolate compared to the others. These other three parasite isolates proved to be sensitive to the five drugs tested (MLC: 0.97-7.81 microg ml).

Animals↗
PubMed2002

The chitinase system from Trichomonas vaginalis as a potential target for antimicrobial therapy of urogenital trichomoniasis.

Chitinolytic activities in Trichomonas vaginalis membrane extracts were assessed by assays of three enzyme systems: N-acetyl-beta-D-hexosaminidase (NAHase), chitobiosidase and chitotriosidase. N-acetyl-beta-D-hexosaminidase was the enzyme that showed the highest specific activity. After successive subcutaneous inoculations into mice and parasite recovery in culture, the enzyme activities increased significantly with the number of inoculations for up to eight passages. In addition, enzyme activities were maximum at the logarithmic phase of growth. Glycol chitin, a chitinase substrate, enhanced all chitinolytic activities by about 30% and a clear-cut correlation is shown between the capacity for erythrocyte lysis by parasites and NAHase expression. Chitobiosidase and chitotriosidase activities were both inhibited at 58% and 100%, respectively, by allosamidine, a chitinase inhibitor used at 3 microM, whereas NAHase activity was not affected. Seven putative NAHase inhibitors (compounds n, 1-7), ureido and thioureido derivatives of 2-amino-2-deoxy-D-glucose were evaluated and five of them had K(i) values in the range 30-70 microM. The most active compound (compound 6) was functionally competitive with respect to the substrate with a K(i) value of 30 microM. The IC(50) values of the most active compounds on T. vaginalis were in the range 62-85 microM. These results indicate that chitinases of T. vaginalis are involved in pathogenicity and they could be an interesting target for drugs since chitinase inhibitors also inhibit parasite growth.

Animals↗
PubMed2001

A historic photomicrograph of a parasite (Trichomonas vaginalis).

Knowing that Alfred Donné was the discoverer of an important human parasite, and finding that he was also a pioneer of photomicrography, it occurred to me that his parasite might well have become a subject of his photography. It was a simple matter to confirm that this was indeed the case. The parasite he discovered was Trichomonas vaginalis; and, in collaboration with Foucault, Donné made a photomicrograph showing several protozoan parasites lying among vaginal epithelial cells. His publication of an engraved image of the photomicrograph in 1845, was a landmark in the history of photography and microbiology.

Animals↗
PubMed1992

Proteinases of Trichomonas vaginalis: antibody response in patients with urogenital trichomoniasis.

Immunoprecipitation combined with electrophoresis in gelatin-polyacrylamide gels was successfully used for detection of antibodies against numerous proteinases of Trichomonas vaginalis in infected patients. The method proved to be highly specific as anti-proteinase antibodies were absent in women with negative cultivation of T. vaginalis and no history of trichomoniasis. Sera of 71% and vaginal washes of 86% patients with trichomoniasis were positive for these antibodies. In vaginal washes, but not in sera, antibodies were partly complexed with proteinases, possibly of trichomonad origin. It was also shown that serum antibodies as well as local anti-proteinase antibodies persisted for weeks after patients had been cured.

Adult↗
PubMed2001

Metronidazole-resistant strains of Trichomonas vaginalis display increased susceptibility to oxygen.

Susceptibility to oxygen and properties relative to oxygen metabolism were compared in metronidazole-resistant and susceptible strains of Trichomonas vaginalis. The study involved clinical isolates displaying the aerobic type of resistance, as well as resistant strains developed in vitro, both with aerobic (MR-3) and anaerobic (MR-5, MR-100) resistance. Elevated sensitivity to oxygen of the resistant clinical isolates was observed. Progressive increase of susceptibility to oxygen also accompanied in vitro development of resistance. No correlation was found between the activity of NADH oxidase and aerobic resistance, while the in vitro derivative with fully developed anaerobic resistance (MR-100) showed about 50% decrease of NADH oxidase activity. The superoxide dismutase (SOD) activity was elevated in both resistant clinical isolates and in in vitro-developed resistant strains. The changes in levels of ferredoxin were insufficient to support ferredoxin deficiency as a cause of aerobic metronidazole resistance. Western blot analysis and electron paramagnetic resonance spectroscopy of purified hydrogenosomes showed that ferredoxin is expressed in aerobically resistant strains and has intact iron-sulfur clusters. Down-regulation of ferredoxin was demonstrated only in the late phase of development of the anaerobic resistance (MR-100). The results support a link between aerobic resistance and defective oxygen scavenging. The increased levels of intracellular oxygen, beneficial to resistant parasites when they interact with the drug, may have adverse effects on their fitness as shown by their increased sensitivity to oxidative stress.

Animals↗
PubMed2006

Nitric oxide radicals in leucocytes and vaginal washes of Trichomonas vaginalis -infected symptomatic and asymptomatic women.

The clinical spectrum of Trichomonas vaginalis infection varies from asymptomatic to mild, moderate or severe vaginitis. Nitric oxide and other reactive nitrogen radicals produced by immune effector cells are important cytotoxic and cytostatic mediators against several microorganisms including parasites. In the present study, inducible nitric oxide synthase (iNOS) and reactive nitrogen intermediates (RNI) were determined in leucocyte cultures (stimulated with T. vaginalis in vitro) and vaginal washes (VWs) of 22 symptomatic and 20 asymptomatic T. vaginalis-infected and 20 healthy women by immunoblotting and Griess method respectively. The iNOS protein was detected in leucocytes and VWs of all the symptomatic and asymptomatic women, but was not detected in any of the samples from healthy women. Mean iNOS protein band intensity was significantly higher in leucocytes as compared to VWs (P<0.001) of both symptomatic and asymptomatic women and was also higher in leucocytes of asymptomatic as compared to symptomatic women (P<0.05). Mean RNI concentration was also significantly higher in leucocytes (P<0.01) and VWs (P<0.05) of asymptomatic as compared to symptomatic women, and was also higher in samples of infected as compared to healthy women (P<0.001). These results suggest that reactive nitrogen radicals may have a role in limiting T. vaginalis infection in asymptomatic women.

Animals↗
PubMed2005

Seroepidemiology of Trichomonas vaginalis in rural women in Zimbabwe and patterns of association with HIV infection.

Serological assays using dried blood spots from 5221 women in rural areas of eastern Zimbabwe were used to assess the epidemiology of Trichomonas vaginalis infection, and its association with HIV. Antibodies to T. vaginalis and to HIV were detected by enzyme immunoassays. Behavioural and demographic data were collected by confidential questionnaires. In total, 516 (9.9%) women were seropositive for T. vaginalis and seroprevalence increased with age among younger women. Divorced, widowed and single women were more likely to be seropositive. After controlling for age, seropositivity was significantly associated with being sexually active, having multiple sex partners, having a partner who had multiple sex partners, and having a new sex partner in the past year. Seropositivity was associated with a recent history of genital discharge. Overall, 208 (40.3%) T. vaginalis-positive samples were also positive for HIV, compared with 1106 (23.5%) T. vaginalis-negative samples (age and sex adjusted OR 2.11, 95% CI 1.74-2.55, P < 0.001). There was increased risk for being HIV-positive amongst T. vaginalis-seropositive women regardless of residence, employment or education. In a logistic regression controlling for common risk factors, the association remained significant. T. vaginalis-seropositive young women with a history of genital discharge were much more likely to be HIV-positive than women who were T. vaginalis-seronegative and had no history of discharge (OR 6.08, 95 % CI 2.95-12.53). Although a causal relationship cannot be assumed, detection and treatment of trichomoniasis may be important in strategies to reduce HIV transmission through sexually transmitted infection control.

Adolescent↗