Molecular identification and phylogenetic analysis of free-living amoeba in the water resources of Arak, Iran.

The aim of the present study was to detect free-living amoeba (FLA) in the water resources of Arak, Iran using molecular tools. A total of 154 samples were collected from different water supplies. Molecular analyses, sequencing, and phylogenetic study were conducted to confirm the species and genotypes of FLA. Fisher exact test was used to determine the significance. Of 154 water samples, 19 (12.3%) samples were tested positive for FLA. Three genotypes of Acanthamoeba including T4, subtype D, and T5 were identified among the isolates. The pathogenicity assay showed that the isolate of Acanthamoeba in drinking water was highly pathogenic. Three species of Naegleria, including N. australiensis, N. pagei, and N. gruberi were found among the samples. Six isolates of Vermamoeba were identified as V. vermiformis. Meanwhile, three other species including Vannella sp., Vahlkampfia avara, and Stenamoeba polymorpha were also recovered from the water samples. Statistical analysis showed a significant difference between the various water resources contaminated with FLA. This is the first study to reveal the presence of S. polymorpha in water sources in Iran. According to the findings of the present study, health officials should be beware of potential public health impacts of FLA in water resources.

Isolation and molecular identification of Acanthamoeba spp. from hospital dust and soil of Khomein, Iran, as reservoir for nosocomial infection.

BACKGROUND: Acanthamoeba spp. are commonest opportunistic amoebae, which ubiquitous in various environmental resources. Acanthamoeba species are the causative agents of amoebic keratitis, granulomatous amoebic encephalitis and i.e. in immunocompromised and immunocompetent patients. Moreover Acanthamoeba spp. can act as reservoir and transmission agent of bacterial pathogens. Due to this issue the aim of this study was to characterized Acanthamoeba spp. genotypes in dust and soil of hospital samples from Khomein of Iran. METHODS: In a cross sectional study, a total of 100 soil and dust samples were collected from hospital environment of Khomein Iran, and analyzed for the presence of Acanthamoeba spp. based on phenotypic and molecular methods including PCR amplification and sequence analysis of 18SrRNA. A total of 5 Acanthamoeba isolates were sequenced, and different genotypes of isolates were detected via direct sequence analysis. RESULTS: The results showed that 20% of samples (20/100) were positive for Acanthamoeba, while only 5 cases were successfully cultured in NNM medium and were subjected to molecular assay. A. lenticulata, A. castellanii and A. quina were the prevalent identified species that were belonged to T4 and T5 genotypes. CONCLUSIONS: Acanthamoeba spp. are the most prevalent free living amoeba in the dust and soil of hospital environment. Moreover, due to the presence of potentially pathogenic T4 genotypes in our hospital, it is recommended that in health and hygienic programs elimination of FLA should be considered.

Bovine cysticercosis in feedlot cattle in central region of Iran.

The cestode, Taenia saginata is a zoonotic tapeworm that it's larval stage which known as Cysticercus bovis cause cyst formation in cattle's organs such as heart, lung, liver, tongue, esophagus and diaphragm muscle, despite the infected cattle may show no clinical signs. Antemortem diagnosis of bovine cysticercosis can be made by antigen detecting ELISA. In a feedlot near city of Arak, beef cattle had different degrees of lethargy, dullness, unthriftiness and were reluctant to move. In postmortem examination of cattle, samples were collected from heart tissue and stained by H&E method for light microscopic examination. 10 ml of blood samples were taken from jugular veins of 90 cattle that were going to be sent to slaughterhouse. Serums obtained from blood samples were investigated for presence of C. bovis antigen by ELISA assay. Soils and dusts from farm yard, pen's floor, feed store and both toilets of workers and employer were sampled and evaluated for presence of parasite eggs by floating method. Cysticercus bovis antigen were identified in serums of 18 cattle; and also, samples from workers toilet was contaminated by eggs of T. saginata. This study showed that serologic methods in conjunction with meat inspection can be used for diagnosis of bovine cysticercosis. The aim of this study is to identify infected cattle with C. bovis by serologic methods before slaughter and determine microscopic characteristics of lesions on postmortem examination in central area of Iran.

Leishmaniasis Caused by Leishmania major on the Glans Penis: A Case Report.

Cutaneous leishmaniasis (CL) is one of the prevalent parasitic diseases in Iran principally caused by two species, Leishmania major and L. tropica. Here, we present a rare case of a congenital form of CL around the glans penis from the central part of Iran in 2017. A 24-yr-old male patient from the central part of Iran presented with biennial ulceration of the glans penis. Diagnostic methods included physical and preclinical examination, microscopic observation, leishmanin skin test (LST), and serological tests including direct agglutination test (DAT). Nested PCR and sequencing analysis were used on the positive smears for confirmation of CL and Leishmania species identification. The preclinical results were normal, and no anti-Leishmania antibodies were detected in the peripheral blood of the patient using DAT. In abdominal ultrasonography, the spleen and liver size were normal. LST was positive (≥5 mm) after 72 h, and a few amastigote forms of Leishmania sp. were demonstrated under light microscopy. L. major was confirmed using nested PCR and sequencing analysis. The patient responded to oral administration of miltefosine (2.5 mg/kg/d) for 28 days. To the best of our knowledge, genital CL due to L. major has not been previously reported from Iran.

Detection of Specific Antibody Reactivity to Toxocara Larval Excretory-secretory Antigens in Asthmatic Patients (5-15 Years).

BACKGROUND & PURPOSE: Humans act as an intermediate host for Toxocara canis and Toxocara cati. Toxocara may be an important risk factor for asthma in humans. The aim of the present study was to evaluate immunoglobulin G (IgG) anti-Toxocara canis antibody, using enzyme-linked immunosorbent assay (ELISA) in asthmatic patients (aged 5-15 years), referring to a clinic of pulmonary diseases in Arak, Iran. MATERIALS & METHODS: In this bi-group cross sectional study, serum samples were collected from 110 children with confirmed asthma and 70 children without asthma within one year. IgG anti-Toxocara antibody was detected viaELISA method. The collected data were analyzed, using SPSS. RESULTS: The seroprevalence of antibodies against Toxocara species was estimated at 1.8% (two males) in asmathic children viaELISA method; however, no antibodies against Toxocara canis were detected in the control group. There was no significant correlation between the frequency of antibodies against Toxocara and variables such as age, gender, or place of residence (P>0.05). Moreover, the frequency of antibodies against Toxocara was not significantly correlated with contact with dogs, consumption of unwashed fruits and vegetables, or use of raw/undercooked sheep liver (P>0.05). CONCLUSION: The present study showed anti-Toxocara antibody in 1.8% of asthmatic children and determined the seroprevalence of toxocariasis in asthmatic children and adolescents in Arak, Iran. Based on the findings, the low rate of infection with Toxocara among asthmatic children may be attributed to acceptable personal hygiene and religious considerations.

Identification of Malassezia Species Isolated from Patients with Pityriasis Versicolor Using PCR-RFLP Method in Markazi Province, Central Iran.

BACKGROUND: The lipophilic yeasts of Malassezia species are members of the normal skin microbial that are cause of pityriasis versicolor. Pityriasis versicolor is a common superficial fungal infection with world-wide distribution. The phenotypic methods for identification of Malassezia species usually are time consuming and unreliable to differentiate newly identified species. But DNA-based techniques rapidly and accurately identified Malassezia species. The purpose of this study was isolation and identification of Malassezia Species from patients with pityriasis versicolor by molecular methods in Markazi Province, Central Iran in 2012. METHODS: Mycologic examinations including direct microscopy and culture were performed on clinical samples. DNA extraction was performed from colonies. The ITS1 region of rDNA from isolates of Malassezia species were amplified by PCR reaction. The PCR were digested by Cfo I enzyme. RESULTS: From 70 skin samples, were microscopically positive for Malassezia elements, 60 samples were grown on culture medium (85.7%). Using PCR-RFLP method, that was performed on 60 isolates, 37(61.6%) M. globosa, 14(23.3%) M. furfur, 5(8.4%) M. sympodialis and 4(6.7%) M. restrictawere identified. In one case was isolated M. globosa along with M. restricta. CONCLUSION: The PCR-RFLP method is a useful and reliable technique for identification of differentiation of Malas-sezia species.

Differential genomics output and susceptibility of Iranian patients with unilocular hydatidose.

BACKGROUND: The objective of this study was to investigate HLA-DRB1*and DQB1* allelic polymorphisms in Iranian patients with hydatidose. This is the first survey dealing with the correlation between HLA-DRB1* and DQB1* alleles and cystic echinococcosis in Iranian patients. METHODS: The study was carried out on 56 patients with confirmed cystic echinococcosis and 30 apparently healthy individuals living in Arak- Markazi Province by HLA-DRB1 and DQB1 typing through PCR-SSP method. The first step was to identify the patients and blood sampling. DNA was prepared from whole blood and PCR-SSP with 31 primer mixes for per sample was used. PCR reaction mixtures were loaded in agarose gels and bands were observed under UV illumination and gel document after electrophoresis. Analysis of results was carried out with specific softwares and frequency and interpretation tables for calculation of P-value in χ(2) test were provided via Fisher's exact test. Significant samples were analyzed by logistic regression and odds-ratios were calculated. RESULTS: A statistically significant positive association was found between HLA-DQB1*03 and the resistance to cystic echinococcosis (P < 0.02) (odds-ratio = 2.87). CONCLUSION: Immunogenetic susceptibility to unilocular hydatidose varies according to the HLA antigens in Arak, Markazi Province, and DQB1*03 molecules are associated with the level of immune response to parasite antigens.

Effects of Toxoplasma gondii Infection in Level of Serum Testosterone in Males with Chronic Toxoplasmosis.

BACKGROUND: Toxoplasma gondii is an intracellular protozoan parasite that infects human and animals. Toxoplasma parasites are isolated from different parts of animals even from semen but there are little information about the effect of toxoplasmosis on fertility in animals and humans. In present study, the effect of chronic toxoplasmosis on serum levels of testosterone in men was studied. METHODS: In this case-control study, 1026 men referred to Arak Post Marriage Center were selected. Three ml of blood samples were collected and sera separated by centrifugation at room temperature. These sera were analyzed for detection of anti-T. gondii IgG antibody. Next 365 positive sera were selected as cases and also the same number of negative sera (365) as controls. Finally the level of testosterone was analyzed for the cases and controls samples. RESULT: Serological tests on the sera of 1,026 men in Arak City showed that 365 of them had anti-Toxoplasma antibody. Comparison of testosterone concentration in case and control groups showed that testosterone concentration in case group was less than control group and this difference was statistically significant (P<0.05). CONCLUSION: The chronic toxoplasmosis could affect reproductive parameters in men.