The Effect of Hydroalcoholic Extract of Quercus brantii and Artemisia aucheri Boiss Against Trichomonas vaginalis In vitro.

Objective: Trichomoniasis is the most common sexually transmitted protozoan infection worldwide. Metronidazole is widely considered as the drug of choice for treating of trichomoniasis but considering its potential side effects, we aimed to assess the therapeutic influences of hydro-alcoholic extracts of Quercus brantii and Artemisia aucheri Boiss as alternative medications against Trichomonas vaginalis (T. vaginalis). Methods: The trophozoites were cultured in TYI-S-33 medium at a density of 5x105 trophozoites/mL. Subsequently, they were incubated with varying concentrations of the plant extracts (32, 64, 125, 250, 500, and 1,000 µg/mL) and metronidazole (16, 32, 64, 125, 250, and 500 µg/mL), as the positive control. The number of trophozoites in each well plate was quantified after 2, 4, 6, 24, 48, and 72 hours using trypan blue staining. Finally, the viability of the parasite was assessed by vital methylene blue staining. Results: The hydro-alcoholic extracts of Q. brantii and A. aucheri Boiss at concentrations of 125, 250, 500, and 1,000 µg/mL demonstrated significant efficacy against the parasite. Our findings indicated that the minimum effective concentrations were 125 µg/mL and hydro-alcoholic extracts of Q. brantii and A. aucheri Boiss have the ability to effectively eliminate T. vaginalis after 48 and 72 hours of treatment. Conclusion: The findings of the present study showed that hydro-alcoholic extract of Q. brantii and A. aucheri Boiss can induce death in T. vaginalis. However, further complementary in vivo studies are needed to assess the components of these plants in the treatment of T. vaginalis.

Molecular Targets Implicated in the Antiparasitic and Anti-Inflammatory Activity of the Phytochemical Curcumin in Trichomoniasis.

Trichomoniasis, is the most prevalent non-viral sexually transmitted disease worldwide. Although metronidazole (MDZ) is the recommended treatment, several strains of the parasite are resistant to MDZ, and new treatments are required. Curcumin (CUR) is a polyphenol with anti-inflammatory, antioxidant and antiparasitic properties. In this study, we evaluated the effects of CUR on two biochemical targets: on proteolytic activity and hydrogenosomal metabolism in Trichomonas vaginalis. We also investigated the role of CUR on pro-inflammatory responses induced in RAW 264.7 phagocytic cells by parasite proteinases on pro-inflammatory mediators such as the nitric oxide (NO), tumor necrosis factor α (TNFα), interleukin-1beta (IL-1ß), chaperone heat shock protein 70 (Hsp70) and glucocorticoid receptor (mGR). CUR inhibited the growth of T. vaginalis trophozoites, with an IC50 value between 117 ± 7 µM and 173 ± 15 µM, depending on the culture phase. CUR increased pyruvate:ferredoxin oxidoreductase (PfoD), hydrogenosomal enzyme expression and inhibited the proteolytic activity of parasite proteinases. CUR also inhibited NO production and decreased the expression of pro-inflammatory mediators in macrophages. The findings demonstrate the potential usefulness of CUR as an antiparasitic and anti-inflammatory treatment for trichomoniasis. It could be used to control the disease and mitigate the associated immunopathogenic effects.

Trichomonas vaginalis NTPDase inhibited by lycorine modulates the parasite-neutrophil interaction.

Lycorine is an Amaryllidaceae alkaloid that presents anti-Trichomonas vaginalis activity. T. vaginalis causes trichomoniasis, the most common non-viral sexually transmitted infection. The modulation of T. vaginalis purinergic signaling through the ectonucleotidases, nucleoside triphosphate diphosphohydrolase (NTPDase), and ecto-5'-nucleotidase represents new targets for combating the parasite. With this knowledge, the aim of this study was to investigate whether NTPDase and ecto-5'-nucleotidase inhibition by lycorine could lead to extracellular ATP accumulation. Moreover, the lycorine effect on the reactive oxygen species (ROS) production by neutrophils and parasites was evaluated as well as the alkaloid toxicity. The metabolism of purines was assessed by HPLC. ROS production was measured by flow cytometry. Cytotoxicity against epithelial vaginal cells and fibroblasts was tested, as well as the hemolytic effect of lycorine and its in vivo toxicity in Galleria mellonella larvae. Our findings showed that lycorine caused ATP accumulation due to NTPDase inhibition. The alkaloid did not affect the ROS production by T. vaginalis; however, it increased ROS levels in neutrophils incubated with lycorine-treated trophozoites. Lycorine was cytotoxic against vaginal epithelial cells and fibroblasts; conversely, it was not hemolytic neither exhibited toxicity against the in vivo model of G. mellonella larvae. Overall, besides having anti-T. vaginalis activity, lycorine modulates ectonucleotidases and stimulates neutrophils to secrete ROS. This mechanism of action exerted by the alkaloid could enhance the susceptibility of T. vaginalis to host immune cell, contributing to protozoan clearance.

Improved subtyping affords better discrimination of Trichomonas gallinae strains and suggests hybrid lineages.

Trichomonas gallinae is a protozoan pathogen that causes avian trichomonosis typically associated with columbids (canker) and birds of prey (frounce) that predate on them, and has recently emerged as an important cause of passerine disease. An archived panel of DNA from North American (USA) birds used initially to establish the ITS ribotypes was reanalysed using Iron hydrogenase (FeHyd) gene sequences to provide an alphanumeric subtyping scheme with improved resolution for strain discrimination. Thirteen novel subtypes of T. gallinae using FeHyd gene as the subtyping locus are described. Although the phylogenetic topologies derived from each single marker are complementary, they are not entirely congruent. This may reflect the complex genetic histories of the isolates analysed which appear to contain two major lineages and several that are hybrid. This new analysis consolidates much of the phylogenetic signal generated from the ITS ribotype and provides additional resolution for discrimination of T. gallinae strains. The single copy FeHyd gene provides higher resolution genotyping than ITS ribotype alone. It should be used where possible as an additional, single-marker subtyping tool for cultured isolates.

Comparative effect of manuka honey on anaerobic parasitic protozoans with standard drug therapy under in vitro conditions: A preliminary study.

BACKGROUND: From the past five decades, metronidazole and tinidazole have been used for treating nonresistant and resistant giardiasis and trichomoniasis. However, due to the occurrence of drug resistance to standard therapy idealizes us to explore some additional therapies which is cost-effective, easy accessibility, and natural which has least side effects. Manuka honey obtained from Leptospermum scoparium is well known for its antibacterial and wound healing properties and is thought to be a better option as an additional therapy. OBJECTIVE: The present study was conducted to find out the effect of manuka honey on anaerobic protozoans that includes Giardia and Trichomonas under in vitro conditions in comparison to metronidazole and tinidazole. MATERIALS AND METHODS: Axenic culture of Giardia lamblia strain Portland 1 and Trichomonas vaginalis strain 413 was used for drug sensitivity assay to tinidazole, metronidazole, and manuka honey with the highest concentration of 17.1 µg/ml, 24.7 µg/ml, and 50%v/v by using (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole). For this, head-to-head comparison has been done and IC 50 of the standard drug as well as manuka honey was calculated. RESULTS: The result showed that percentage inhibition on the growth of both the parasites is dependent on concentration as well as exposure time of the drug. The calculated IC 50 was found to be 5.6%v/v and 1.5%v/v for manuka honey with respect to G. lamblia and T. vaginalis. CONCLUSION: The present study suggests that manuka honey can be used as an additional therapy for the patient with giardiasis or trichomoniasis. However, in vivo study in the near future will elucidate more about the effectiveness of honey in treating parasitic infections.

Comparison of in vitro activity of metronidazole and garlic-based product (Tomex®) on Trichomonas vaginalis.

Trichomonas vaginalis is a common sexually transmitted parasite in humans. Metronidazole has been the gold standard for treatment of trichomoniasis. The prevalence of metronidazole resistance and its unpleasant adverse effects drew the attention to the investigation of other lines of treatment, as that of herbal medicine. Garlic has been proven to have antibacterial, antiprotozoal, and antihelminthic activity. The current study was carried out to evaluate the efficacy of commercially available garlic (Tomex®) on T. vaginalis in vitro. The effect of different concentrations of garlic (12.5, 25, 50, and 100 µg/ml) was determined on multiplication and motility of trophozoites at different time points (after 24, 48, 72, and 96 h) in comparison to the same concentrations of metronidazole at the same different time points. The results showed that parasite multiplication inhibition was noticed in proportion of concentration of Tomex and incubation time. The minimal lethal concentration of Tomex was 100 µg/ml after 24 h, 50 µg/ml after 48 h, 25 µg/ml after 72 h, and 12.5 µg/ml after 96 h. These results were similar to that of metronidazole as its minimal lethal concentration was 50 µg/ml after 24 and 48 h and 12.5 µg/ml after 72 and 96 h. Garlic had completely inhibited the motility of trophozoites with concentration of 100 µg/ml after 24 h, 50 µg/ml after 48 h, 25 µg/ml after 72 h, and 12.5 µg/ml after 96 h while metronidazole had completely inhibited the motility of trophozoites with concentration of 50 µg/ml after 24 h, 25 µg/ml after 48 h, and 12.5 µg/ml after 72 and 96 h. This suggests that commercially available garlic (Tomex®) may be a promising phytotherapeutic agent for trichomoniasis.

In vitro effects of various plants extracts on the growth of Trichomonas vaginalis.

Trichomoniasis is a common sexually transmitted disease (STD) caused by a protozoan parasite called Trichomonas vaginalis. This disease, with roughly 170 million new infected people worldwide per year, is associated with various problems such as pre-term delivery, high infant mortality or low birth weight. In addition, trichomoniasis increases patient susceptibility to HIV infection. The mainstay medication for trichomoniasis is metronidazole, but some resistant strains to this treatment have been identified. Moreover, treatment with metronidazole is associated with numerous side effects. Thus efforts to identify new alternative drugs in order to control trichomoniasis are vital. The use of medicinal herbs has gained widespread acceptance in both developing and non-developing societies because of owing to fewer side effects and better patient tolerance. In our search for alternative drugs in the treatment of trichomoniasis, we reviewed the effect of different plant extracts on Trichomonas vaginalis in vitro.

Modulation by Polypodium leucotomos extract of cytokine patterns in experimental trichomoniasis model.

The immunomodulating effects of Anapsos, an aqueous hydrosoluble extract obtained from the rhizomes of the fern Polypodium leucotomos, on both pathogenicity and cytokine levels in serum (IFN-gamma/IL-4) were assayed in a Trichomonas vaginalis experimental model (BALB/c mice infected with 10(7) trichomonads and examined at day 15 after infection). Doses of 20 mg/kg/day administered for 10 days before the infection with the parasite induced a decrease of the experimental pathogenicity approximately 10-20% compared to controls. Gross histopathologic changes at abdominal organs and mortality rate, as a consequence of pathogenicity of the protozoa and the immune response of the host, were evaluated. IFN-gamma and IL-4 cytokines were determined on days -5, 0, 5, 10, and 15 postinfection by indirect ELISA. Treatment with PAL before infection modulates and downregulates the IFN-gamma concentration, while anticipates and upregulates the IL-4 level. The assays performed have showed the utility of the murine model of experimental trichomoniasis for the evaluation of immunomodulatory activity of synthetic or natural products.

Antitrichomonal action of emodin in mice.

Emodin, an active component contained in the root and rhizome of Rheum palmatum L. (Polygonaceae), was found to have an inhibitory effect on the pathogenicity of Trichomonas vaginalis in mice. Emodin delayed the development of subcutaneous abscesses due to infection of this parasite. Also, it cures the intravaginal infection of trichomonads through oral administration. In cell cultures, it reduced the cytotoxic effect of this parasite towards mammalian cells. This inhibition was markedly reversed by the coexistence of free radical scavengers, indicating the possible mediation of free radicals.