Glia Maturation Factor Beta: A Novel Neuro-Impairment Prediction Factor in Toxoplasmosis.

Background: Toxoplasma gondii, a neurotropic protozoan, infects up one to third of the world population. The parasite can invade a wide variety of nucleated cells but preferably glial cells. Glia maturation factor ß (GMFß), a 17KD protein expressed at high levels in the central nervous system is predominantly related to neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Multiple sclerosis. We aimed to determine the expression level of GMFß and its relation to other pro-inflammatory factors (IL33, SDF1, and CCL2) on T. gondii infected human neuroblastoma cell line. Methods: The human neuroblastoma (SK_NMC C535) cell line was infected by 5×106 (1:1 ratio). The supernatant was collected after cell lysis and centrifugation. Total RNA was extracted using the Yekta Tajhiz RNA extraction kit. cDNA was synthesized based on RevertAid First Strand cDNA Synthesis Kit manufacturer's protocol (Parstous, cDNA synthesis kit, Iran). The specificity of each primer pair (GMFß, IL33, SDF1, and CCL2) was provided by NCBI BLAST. Gene expression level was measured using Real-Time PCR. All experiments were conducted at the Hamadan University of Medical Sciences, western Iran in 2022. Results: The GMFß increased significantly up to 1.35-fold (P=0.007). The increase in GMFß expression in neuroblastoma cells was consistent with the increase in pro-inflammatory factors (CCL2 (0.47), IL33 (0.152) and, SDF1 (1.33)). Conclusion: GMFß upregulation can be a novel indicator of the destruction of nerve cells.

Alpha-Pinene and Tannic Acid Inhibit Trichomonas vaginalis Protozoan Cells by Inducing Apoptosis.

Background: Trichomoniasis is one of the most common sexually transmitted infections worldwide. The growing concern of drug resistance of this infection has cautioned the need for new drug development. We evaluated the potential antiproliferative and apoptotic effect of α-pinene and tannic acid (TA) on Trichomonas vaginalis cells. In addition, the cytotoxicity of agents on Vero cells was investigated. Methods: Trichomonas cells were axenically cultured in TYI-S-33 medium. In vitro antiproliferative activity of α-pinene, TA, and metronidazole was investigated against Trichomonas cells. The assays were carried out in triplicate using microtiter plate and trypan blue staining method. Annexin V/PI staining with flow cytometry was used to evaluate apoptosis induction. In addition, the cytotoxic effect was measured by MTT assay. Results: α-Pinene and TA exhibited significant inhibition of the Trichomonas cells and the lowest IC50 values were 22.9 µg/ml and 140 µg/ml at 48 hours' incubation, respectively. The CC50 was found at 116 µg/ml for α-pinene and 473 µg/ml for TA, after 48 hours of treatment. The flow cytometry study demonstrated that the natural compounds induced apoptosis in Trichomonas cells. After 24 hours of treatment, the induction of apoptosis was 5.2% - 36.6% at concentrations of 3.9 - 62.5 µg/ml for α-pinene and TA induced-apoptosis was 6.1% - 53.8% at concentrations of 125-2000 µg/ml. Conclusion: Although the results show the antiproliferative and apoptotic effect of α-pinene and TA on Trichomonas cells, in vivo studies are needed to further clarify the effects of these compounds.

Genotyping of Environmental Isolates of Acanthamoeba in Hamadan, West of Iran.

Background: Different genotypes of Acanthamoeba have been abundantly isolated in environmental samples such as water, soil, and dust, as well as in different hospital departments and eyewash stations. This protozoan is a potential hazard for immunocompromised patients and contact lens wearers. The aim of the present study was isolation and genotyping of environmental and corneal isolates of Acanthamoeba in Hamadan, west of Iran. Methods: During 2018-2020, a total of 104 environmental samples including, water, soil, and dust and 16 corneal scraping samples were collected and investigated for the presence of Acanthamoeba using morphological and molecular identification tools. Genotypes were determined using sequence analysis of the diagnostic fragment 3 (DF3) from Acanthamoeba-specific amplimer S1 (ASA.S1) gene. Phylogenetic tree was constructed with the MEGA7 software using Neighbor-Joining method. Results: The presence of Acanthamoeba spp. was determined in 87.5% of water, 53.1% of soil, and 25% of dust samples. From 30 dust samples collected from eight wards of three hospitals, 7 (23.3%) were contaminated with Acanthamoeba. Sequencing analysis of environmental samples revealed that the T4 genotype was the most prevalent (92.6%) one. Genotypes T2 (1.9%), T2/T6 (1.9%), and mixed T4 and T2/T6 (3.7%) were also identified in environmental samples. Acanthamoeba was seen in none of the examined corneal scraping samples from patients with suspected keratitis. Conclusion: The widespread occurrence of this potentially pathogenic amoeba in most hospital wards and environmental resources and areas of the region highlights a strong need to increase awareness regarding this ubiquitous amoeba among susceptible individuals, such as immunocompromised patients and contact lens wearers.

In vitro antitrichomonal activity of Satureja khuzestanica and main essential oil components carvacrol, thymol, and eugenol.

INTRODUCTION: Human trichomoniasis is a widespread sexually transmitted disease and the concern of drug resistance in the parasite is growing. Hence, this study was performed to evaluate in vitro antitrichomonal activity of Satureja khuzestanica, carvacrol, thymol, eugenol, and phytochemical evaluation of the S. khuzestanica oil. METHODOLOGY: Extracts and essential oil of S. khuzestanica, and the components were prepared. Then, susceptibility testing was performed using the microtiter plate method and Trichomonas vaginalis isolates. The minimum lethal concentration (MLC) of the agents was determined in comparison with metronidazole. Also, the essential oil was investigated by gas chromatography-mass spectrometry and gas chromatography-flame ionization detector. RESULTS: After 48 hours of incubation, carvacrol and thymol were the most effective antitrichomonal agents with MLC of 100 µg/mL, followed by the essential oil and hexanic extract (MLC = 200 µg/mL), then eugenol and methanolic extract (MLC = 400 µg/mL), in comparison with the metronidazole MLC of 6.8 µg/mL. Overall, 33 identified compounds accounted for 98.72% of the total essential oil composition with carvacrol, thymol, and p-cymene being the major constituents. CONCLUSIONS: The results suggested the potency of S. khuzestanica and its bioactive ingredients against T. vaginalis. Thus, further in vivo studies are required to evaluate the efficacies of the agents.

Deciphering the role of miR-71 and let-7 in the fertility of cystic echinococcosis cysts: a preliminary assessment.

Cystic echinococcosis (CE) is a neglected helminthic zoonosis in many parts of the world. Some CE cysts in the intermediate host are non-fertile. Considering the function of microRNAs in many biological processes such as embryonic development, cell proliferation, and apoptosis, this study investigated the function and comparison of miR-71 and let-7 in fertile and non-fertile CE cysts. Here, we determined the expression level of the miRNAs for 33 animal cysts and 16 human cysts (Echinococcus granulosus sensu stricto (G1). The quantitative real-time PCR method was conducted for the expression evaluation of miR-71 and let-7. The expression of both miRNAs in all samples was determined using the following formula: [ΔCT = CT (target) - CT (internal control)]. A comparison of Δct of miR-71 and let-7 in fertile and non-fertile cysts did not show a significant difference (P = 0.911 and 0.354). In cattle, sheep, and humans, Δct of miR-71, and let-7 were higher, respectively. Therefore, the mean expression of miR-71 and let-7 indicates an increase in humans compared to other intermediate hosts. Also, statistical results show a significant difference in the expression of these miRNAs in sheep, cattle, and human cysts (P = 0.025 and 0.01). The lower expression of these miRNAs in cattle cysts and their common infertility might be associated with the hypothesis and function of miRNAs in the fertility of CE cysts. So we should not ignore the function and role of miRNAs in this subject due to the importance of infertility in E. granulosus epidemiology.

Design of highly sensitive nano-biosensor for diagnosis of hydatid cyst based on gold nanoparticles.

Cystic echinococcosis, a zoonotic parasitic infection, is a major public health and economic concern, with worldwide distribution. The development of sensitive diagnostic methods for hydatid disease is important. We designed a highly sensitive nano-biosensor for the diagnosis of hydatid cyst based on gold nanoparticles (AuNPs). AuNPs were synthesized. Echinococcus granulosus antigen was coated on the ELISA microwells. Then, the E. granulosus IgG antibody was added to the microwells. After incubation and washing, the Ag-Ab complex was incubated with a human IgG HRP​-conjugated antibody. Then, the synthesized AuNPs and tetramethylbenzidine (TMB), as a chromogenic substrate of HRP, were added to the reaction. Finally, the absorption rate was measured by spectrophotometry. The results showed that the enzyme peroxide and TMB change the color of the reaction from red to yellow by oxidizing AuNPs. The sensitivity and specificity of the designed method were investigated. The linear equation and regeneration of nanobiosensor designed for red color Y = 0.0312X + 0.649, R2 9962 and for yellow color Y = 0.013X + 0.398, R2 9851 were determined. The limit of detection of the designed nanobiosensor was 0.001 µg mL-1. The results confirmed that the designed nanobiosensor was completely specific for the detection of E. granulosus antibody.

Prevalence of toxoplasmosis in patients infected with tuberculosis; a sero-molecular case-control study in northwest Iran.

In this study, we investigated the possible association between TB and Toxoplasma gondii infection. One hundred confirmed TB individuals living in northwest Iran were classified into three subgroups; newly diagnosed patients (NTB), old diagnosed patients (OTB) and multidrug resistance patients (MDR-TB). One hundred healthy subjects in the same age and sex distribution were ethnically matched. Sera samples were screened for anti-Toxoplasma antibodies. Nested-PCR was performed by targeting the B1 and GRA6 genes. The frequency of Toxoplasma infection based on IgG titer was 71.1% in the OTB subgroup and 33% in the control group, indicating significant association between Toxoplasma seropositivity and OTB (P = 0.001). According to phylogenetic network, the type I strain of Toxoplasma was identified in the OTB subgroup (10.1%). We concluded that patients with OTB subgroup are at high risk for acquisition of Toxoplasma infection which could reactivate the latent toxoplasmosis.

Intestinal parasitic infection among mentally handicapped students in the Islamic Republic of Iran.

BACKGROUND: Intestinal parasitic infections are among the most common public health problems in populations with poor hygiene. Consequently, mental retardation increases the risk of infection. AIMS: The aim of this study was to investigate the prevalence of intestinal parasitic infections among mentally disabled young people in Hamadan, western Islamic Republic of Iran. METHODS: This descriptive cross-sectional study was conducted on 318 students in 9 special schools in 2017. Triplicate faecal specimens for each student were subjected to stool analysis using direct wet mount, sedimentation concentration, and permanent staining technique. Chi-squared or Fisher's exact test were used for statistical analysis. RESULTS: Out of 318 students, 135 (42.5%, 95% CI: 37.1-47.9) were infected with intestinal parasites. Prevalence rates for females and males were 45.2% (61/135) and 40.4% (74/183) respectively. Protozoan infections (42.5%, 135) were more numerous than helminthiasis (0.6%, 2). Blastocystis hominis (28%, 89) was the most common parasite, followed by Giardia lamblia (9.7%, 31), Entamoeba coli (9.1%, 29) and Trichomonas hominis (6%, 19). CONCLUSIONS: The high prevalence rate of intestinal protozoan infection reflects poor personal hygiene among the students and inappropriate environmental conditions. Therefore, specific public health measures are required for the mentally handicapped students in special schools.

Molecular Epidemiologic Study of Male Trichomoniasis in Hamadan, Western Iran.

BACKGROUND: Trichomoniasis, caused by Trichomonas vaginalis protozoan, may lead to clinical or subclinical urethritis or prostatitis in men. Despite the importance of men in the epidemiology of trichomoniasis, there is little information about this topic. This epidemiological study was performed on men in Hamedan, western Iran. METHODS: During Oct 2018 to Mar 2019, 214 male individuals, presenting to the Urology Clinic of Shahid Beheshti Hospital in Hamadan, were enrolled and evaluated for trichomoniasis. First-voided urine specimen was used for detection of T. vaginalis infection using molecular and parasitological methods. RESULTS: Trichomoniasis was detected in 10 of 214 male participants (4.7%, 95% CI: 7.5-1.8%) using PCR assay. Culture and wet mount preparation of urine sediment were unable to isolates any T. vaginalis parasite. Nine of the 10 infected men were married, and six of them were ≥49 yr of age. Urinary frequency and dysuria were the most complaints (80%) among infected individuals. CONCLUSION: Given the notable prevalence of the infection, the prevalence of male trichomoniasis will be underestimated if only conventional diagnostic methods are used. Therefore, the risk of infection as well as the molecular survey of T. vaginalis infection should be considered in men with or without clinical symptoms.

Genotyping of Fresh and Parafinized Human Hydatid Cysts Using nad1 and cox1 Genes in Hamadan Province, West of Iran.

BACKGROUND: Hydatidosis is a cosmopolitan zoonotic infection and Hamadan Province in the west of Iran is one of the most important foci of human hydatidosis in Iran. The aim of the current study was the genetic characterization of hydatid cysts operated from humans in Hamadan Province. METHODS: Seventy-two hydatid cysts samples including 50 paraffinized and 22 fresh human hydatid cysts collected from different hospitals in Hamadan Province, western Iran. The cysts' DNA genome was extracted by kit and PCR was performed for amplifying the fragments of 400 and 450bp for nad1 and cox1 mitochondrial genes, respectively. Genotype diversity and sequence variations of the cysts' isolates were studied by related software. RESULTS: DNA from all (100%) paraffinized and fresh hydatid cysts samples extracted successfully. All paraffinized and fresh hydatid cysts samples were amplified by PCR assay using nad1gene, however, only 18 and 8 samples from paraffinized and fresh hydatid cyst samples was amplified using cox1 gene, respectively. The sequences analysis indicated that, 98.61% the Echinococcus granulosus samples were belong to the genotype G1 and 1.39% were G3 genotype. CONCLUSION: Genotypes of E. granulosus in human samples in Hamadan Province are G1 and G3 and these findings are proved by phylogenic analysis.

Toxoplasma gondii: A Possible Inducer of Oxidative Stress in Reproductive System of Male Rats.

BACKGROUND: Toxoplasmosis is suspected to have adverse effects on the male reproductive system. We aimed to determine the possible role of Toxoplasma gondii in oxidative stress in reproductive system of male rats. METHODS: This study was performed from 2018 until 2019 at the Parasitology Research Laboratory of Hamadan University of Medical Sciences, Hamadan, Iran. Eighty male Wistar rats were randomly divided to control and test groups. The animals in the test group were inoculated by 107 T. gondii RH strain tachyzoites and the control group were injected by 0.2 ml of phosphate-buffered saline. The both groups were following every 10 days until day 80 post inoculation. Oxidative stress markers (OSMs) including antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and nonenzymatic markers including malondialdehyde (MDA), reduced glutathione (GSH) and total antioxidant capacity (TAC) were assessed in testis and serum of T. gondii infected rats. RESULTS: After post inoculation, the variations of the OSMs in the testis tissue of infected rats were as follows: a significant decrease of SOD on day 80 (P=0.03), and CAT activity were detected on day 60 and 80 (P=0.04 and P=0.01) respectively. In addition, GSH (P =0.01) and TAC (P =0.03) concentration were significantly reduced on day 80. On the contrary, the concentration of MDA (P =0.01) was increased 70 days after infection. In addition, consistent changes with the tissue testis were observed in the serum OSMs of infected rats. CONCLUSION: T. gondii infection caused oxidative stress in testis tissue. Thus, the adverse effects of oxidative stress may affect the male rat reproductive system.

Genotyping of Echinococcus granulosus isolates from livestock based on mitochondrial cox1 gene, in the Markazi province, Iran.

Hydatidosisis a parasitic disease caused by the larval stage of Echinococcus granulosus with different genotypes, and major complications in vital organs such as liver, lungs and, brain. Also, this parasite can infect animals and cause economic damages. Recently, some investigations indicated that the genetic variation of the parasite affects the antigenic, immunogenic and pathogenic features. Therefore, present study conducted to genotyping of the E. granulosus larva based on mitochondrial cox1 gene in livestock in the endemic areas of Markazi province, Iran. In this study, 49 hydatid cysts samples collected from 36 sheep, 11 goats and 2 cattle from different slaughterhouses of Markazi province in central part of Iran, 2017. The mitochondrial cox1 gene was amplified and genotyping were accomplished using sequence analysis. The sequencing analysis indicated that the main genotype G1 (61%) and G3 (37%) were identified. Also, one of the samples shows similarity with the G2 (2%) genotype. The results showed the statistically significant differences between the genotypes in different livestock (P < 0.05). This study indicated that the main genotypes of E. granulosus in Markazi province are G1 and G3 which are related to dog/sheep strain. Therefore, parasite control in dogs and sheep can reduce the risk of transmission of infection to humans.

Giardia lamblia assemblages A and B isolated from symptomatic and asymptomatic persons in Hamadan, west of Iran.

Giardia is a very abundant organism bringing about diarrhoea in human beings. The focus of this analysis was the detection of Giardia lamblia assemblages in human stool specimens in Hamadan, west of Iran, as well as the association between obtained assemblages and clinical symptoms. Faecal samples of 4066 individuals admitted to the medical and health care facilities in Hamadan were inspected microscopically for the existence of Giardia cysts/trophozoites, and the clinical symptoms of the patients were recorded. The DNA of positive samples was isolated from and the nucleotide sequences of both glutamate dehydrogenase (gdh) (n = 15) and triose phosphate isomerase (tpi) (n = 8) genes were analyzed. In direct microscopy, a total of sixty-four samples (1.6%), were considered as positive for G. lamblia cysts or trophozoites. The sequence analysis showed that 18 out of 23 sequenced isolates (78.2%) were assemblage A and 5 (21.7%) were assemblage B. Clinical symptoms were observed in 44.4% and 40% of patients with assemblages A and B, respectively. Overall, the predominant assemblage A detected in the tested samples along with bioinformatics analysis suggest a potential zoonotic transmission in the region of the study. Although advanced analyses are necessary to understand the foundation and path of the infection, it seems that more sanitary regulations regarding contact with livestock and pet animals are essential.

Echinococcus granulosus sensu stricto in Livestock and Human in Hamadan, Western Iran.

BACKGROUND: Cystic echinococcosis, a major public health and economic concern, is a zoonotic helminth infection with worldwide distribution. This study was conducted to investigate the genetic characteristics of hydatid cysts isolated from human and livestock in Hamadan region, western Iran during 2016-2017. METHODS: Ten human hydatid cysts and 40 animal hydatid cysts including 32 sheep, 5 cattle and 3 goats were genotyped by PCR amplification of two mitochondrial genes, cox1 and nad1. Genetic identification of the isolates was performed by using bioinformatics software and mtDNA nucleotide sequences of the parasite, available in GenBank database. RESULTS: The PCR amplification was successfully carried out on 50 hydatid cyst isolates and then the nucleotide sequencing was conducted. The sequence analysis of the samples found that the isolates belonged to E. granulosus sensu stricto including G1 (42/50, 84%), G2 (4/50, 8%) and G3 (4/50, 8%) genotype. The G1 genotype was detected in human (8/10, 80%), sheep (26/32, 81%), cattle (5/5, 100%) and goat (3/3, 100%) hydatid cysts. The G2 and G3 genotypes were found only in sheep and human isolates. Alignment analysis of the cox1 and nad1 gene sequences revealed thirteen and ten sequence types, respectively. CONCLUSION: G1 was the prevailing genotype of E. granulosus in the area and dog-sheep transmission cycle should be considered when implementing hydatidosis control programs. In addition, high genetic diversity was detected among the hydatid cyst isolates.

In Vitro Activity of Foeniculum vulgare and Its Main Essential Oil Component Trans-Anethole on Trichomonas vaginalis.

BACKGROUND: Trichomoniasis is one of the most common nonviral sexually transmitted infections worldwide which drug-resistant cases of the infection are rising. The aim of the study was to assessment the in vitro activity of Foeniculum vulgare and its main essential oil component on Trichomonas vaginalis. Also phytochemical investigation of F. vulgare essential oil was performed. METHODS: Five T. vaginalis isolates subjected to susceptibility testing against essential oil and extracts of F. vulgare and anethole using microtiter plate method. The minimum lethal concentration (MLC) of the natural products was assessed in comparison with metronidazole. Gas chromatography-mass spectrometry and gas chromatography-flame ionization detector was applied for chemical investigation of the essential oil. RESULTS: After 48 hours incubation, the most potent antitrichomonal agents were the methanolic and hexanic extract with MLC of 360 µg/ml and followed by the essential oil and anethole (1600 µg/ml). The isolates were sensitive to metronidazole with a mean MLC of 13.7 µg/ml. E-Anethole (88.41 %) was the major constituent of F. vulgare essential oil. CONCLUSION: The results suggested in vitro antiprotozoal properties of F. vulgare and anethole against T. vaginalis. Therefore further studies are needed to evaluate their in vivo effects and toxicity.

Genetic Identification of Echinococcus granulosus Isolates in Hamadan, Western Iran.

BACKGROUND: Cystic echinococcosis is a zoonotic infection and considered as a major economic and public health concern worldwide. This research was conducted to determine genotypic characteristics of livestock and human hydatid cyst isolates from Hamadan area, western Iran. METHODS: Sampling was conducted in Hamadan industrial slaughterhouse and Beast Hospital of Hamadan City, western Iran, from 2015 to 2016. Overall, 74 livestock isolates including 69 sheep, 3 cattle and 2 goats and 9 human hydatid cysts were genotyped by PCR amplification of the rDNA ITS1 region and followed restriction fragment length polymorphism (RFLP) analysis with four restriction endonuclease enzymes, RsaI, HpaII, AluI, and TaqI, and sequencing. RESULTS: The PCR amplicon size of each isolate was approximately 1 kb which was the same with that of sheep strain. According to the RFLP patterns, the isolates belonged to a single species, E. granulosus sensu stricto (G1-G3 complex). Furthermore, sequencing of representative amplicons confirmed that the RFLP-genotyped isolates corresponded to E. granulosus sensu stricto. CONCLUSION: E. granulosus sensu stricto is the prevailing species of E. granulosus sensu lato in the region and pointed out the importance of sheep/dog cycle in human transmission.

Prevalence of Animal Fasciolosis and Specification of Fasciola spp. Isolated from Sheep, Goats and Cattle by Molecular Method: Hamadan Province, West of Iran.

BACKGROUND: Fascioliasis is a common disease among humans and animals. Having global distribution, disease is developed by hepatic trematodes, Fasciola hepatica and F. gigantica. The main objective of this research was determining the prevalence of Fasciola species in Hamadan livestock and identifying those using PCR-RFLP. METHODS: Overall, 13607 livestock livers in the slaughterhouse of Hamadan, west of Iran including 10846 sheep, 995 cattle and 1766 goats were examined in 2015. In addition, 75 Fasciola (41 worms from sheep, 22 worms from goats and 12 worms from cattle) were examined by PCR-RFLP method. RESULTS: Totally, 100 livers were infected to Fasciola species (total prevalence: 0.74%; sheep 0.5%, goats 1.4%, cattle 1.5%). In the molecular results, prevalence of F. hepatica was higher than (92.5% in the sheep) F. gigantic, and in the cattle 91.5% and in the goats was 54.5% F. hepatica. Genotyping identified species confirmed intermediated types as F. hepatica. CONCLUSION: Prevalence of Fasciola in this province is not so high. Intermediate types identified by PCR-RFLP method determined as F. hepatica by nucleotide sequencing. Because of morphological differences and interspecies variety, the accurate identification of Fasciola species needs using nucleotide sequencing.

Genotyping, Drug Susceptibility and Prevalence Survey of Trichomonas vaginalis among Women Attending Gynecology Clinics in Hamadan, Western Iran, in 2014-2015.

BACKGROUND: In spite of sufficient knowledge about phenotypic variation of Trichomonas vaginalis, its genetic characteristics are poorly understood. We carried out a molecular epidemiology study in which in vitro metronidazole susceptibility of T. vaginalis isolates was considered. METHODS: This study was conducted on 862 women admitted to Gynecology Clinics in Hamadan, west of Iran, during 2014-2015. After recording the socio-demographic and clinical characteristics of participants, vaginal swab samples were taken and subjected to microscopic examination, culture, in vitro sensitivity testing and PCR-restriction fragment length polymorphism (RFLP) analysis. RESULTS: T. vaginalis was detected in 1.9% (16/862) of the samples using two parasitological methods. The all T. vaginalis isolates that subjected to drug susceptibility analysis were sensitive to metronidazole with MICs ranged from 0.4 to 12.8 µg/ml. T. vaginalis genotyping by using actin gene and PCR-RFLP analysis identified three actin type; A (9, 56%), I (6, 38%) and E (1, 6%). No significant correlation was observed between actin genotypes and their clinical manifestation (P>0.05). CONCLUSION: The prevalence of T. vaginalis infection is not noticeable in the region and the most of isolates are hypersensitive to metronidazole. Further studies are needed to clarify the efficiency of the actin gene, as a reliable genetic marker, for molecular epidemiology of trichomoniasis.

Evaluation of interleukin 8 +2767 A/T polymorphism in visceral leishmaniasis.

OBJECTIVE: To evaluated the relationship between the genetic variations at IL-8 +2767 position with VL pathogenesis among Iranian patients. METHODS: Three groups including patients with VL clinical presentation and leishmania seropositive (n = 124), patients seropositive but without clinical presentation (n = 82) and healthy controls (n = 63) were selected to conduct this cross-sectional study. Polymorphism at +2767 position of IL-8 was investigated using PCR-RFLP techniques. Anti-leishmania antibody titration was evaluated by the immunoflorescence technique. RESULTS: We observed higher significant frequencies +2767 A/A and A/T genotypes in Group 1 compared to Group 2 and healthy controls (P = 0.001). Also, patients in Group 1 carrying A/A genotype showed higher titer of anti-leishmania antibody than patients with A/T and T/T genotypes (P = 0.05). The validity of the data was analyzed using Hardy-Weinberg equilibrium and one way analysis of variance (ANOVA), as well as χ2 tests. CONCLUSIONS: Our findings indicate that the IL-8 +2767 polymorphism is significantly involved in impaired immune responses against VL and it could be considered as a risk factor for the VL progress.

In Vitro Susceptibility of Iranian Isolates of Trichomonas vaginalis to Metronidazole.

BACKGROUND: Metronidazole, a 5-nitroimidazole derivative, is the main antitrichomonal agent of choice for treatment of trichomoniasis. Since 1962, some cases of treatment failure with metronidazole have been reported and recently drug resistance is now on the rise in the world. This study was aimed to determine current susceptibility of Iranian isolates of Trichomonas vaginalis to metronidazole. METHODS: This study was performed on 50 T. vaginalis isolates collected from west and central areas of Iran. After axenisation of the parasites, susceptibility testing was carried out by using serial twofold dilutions of metronidazole (400 to 0.1 µg/ml). The minimum inhibitory concentration (MIC) and the minimum lethal concentration (MLC) of the trichomonads were determined after 48 h incubation at 35.5 °C. Drug susceptibility assays of the all isolates were carried out two times in triplicate under aerobic and anaerobic conditions. RESULTS: Ninety-eight percent of the T. vaginalis isolates (49/50) were sensitive to metronidazole. Metronidazole resistance was defined as aerobic MIC ≥50 µg/ml, detected in one isolate. The means of aerobic MICs and MLCs and that of anaerobic MICs of the parasites were 2.91, 1.95 and 0.28 µg/ml, respectively. CONCLUSION: This investigation showed in vitro low-level tolerance to metronidazole in a few T. vaginalis isolates that may be leading to the development of drug resistance.