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 TaqΙ, 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

Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates

The majority of the keratitis-causing Acanthamoeba isolates are genotype T4. In an attempt to determine whether predominance of T4 isolates in Acanthamoeba keratitis is due to greater virulence or greater prevalence of this genotype, Acanthamoeba genotypes were determined for 13 keratitis isolates and 12 environmental isolates from Iran. Among 13 clinical isolates, eight (61.5 %) belonged to T4, two (15.3 %) belonged to T3 and three (23 %) belonged to the T2 genotype. In contrast, the majority of 12 environmental isolates tested in the present study belonged to T2 (7/12, 58.3 %), followed by 4/12 T4 isolates (33.3 %). In addition, the genotypes of six new Acanthamoeba isolates from UK keratitis cases were determined. Of these, five (83.3 %) belonged to T4 and one was T3 (16.6 %), supporting the expected high frequency of T4 in Acanthamoeba keratitis. In total, the genotypes of 24 Acanthamoeba keratitis isolates from the UK and Iran were determined. Of these, 17 belonged to T4 (70.8 %), three belonged to T2 (12.5 %), three belonged to T3 (12.5 %) and one belonged to T11 (4.1 %), confirming that T4 is the predominant genotype (S2 = 4.167; P = 0.0412) in Acanthamoeba keratitis

Aeromycological analysis of allergenic airborne fungi in Qazvin, Iran

Background and Purpose: Airborne fungi are one of the most important agents responsible for triggering allergic reactions such as rhinitis and severe asthma. This study was conducted to analyze and monitor the prevalence and distribution patterns of atmospheric fungal aerosols in the air of Qazvin during winter of 2012. Materials and Methods: In the current descriptive study, the incidence and diversity of potentially allergenic airborne fungi were determined using two times sampling interval in 25 different locations of Qazvin city by Petri dish trapping technique and exposure of 10- cm diameter plates of Sabouraud’s dextrose agar medium plus chloramphenicol to the air. Results: A total of 2867 fungal colonies were counted on 156 Petri dishes. Of the identified 18 microfungi genera, Cladosporium spp. was the most frequently isolated genera representing 30.9% of isolates, followed by 30.9% Penicillium spp. (27.3%), Aspergillus spp %) . (24.5 , Alternaria spp. (3.3%), Rhizopus spp. (3.1%), and other fungal genera. Conclusion: The high prevalence, high quantity and variety of allergenic airborne fungi in the air of Qazvin showed that people residing in this area are exposed to health hazards. Furthermore, reduction of exposure to bio-aerosols containing these outdoor fungi is necessary to improve the health of individuals, especially those sensitive to fungal-induced diseases like asthma

In-Vitro Efficacy of Plantago lanceolata L. Extracts on Trichomonas Vaginalis

Abstract Background: Trichomoniasis is one of the most common non viral sexually transmitted diseases worldwide. The aim of this study was to investigate the effect of Plantago lanceolata extracts on Trichomonas vaginalis. Materials and Methods: In this study, after collection and drying of P. lanceolata, n-hexanic, ethyl acetate, methanol and hydroalcoholic extracts, they were prepared by maceration. Five clinical T. vaginalis isoleates subjected to extract suscebtibility testing, in comparison of metronidazole. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) tests were carried out in duplicate and repeated two times for each T. vaginalis isolate. Results: The results showed that the extracts of P. lanceolata had potent antitrichomonal activity. The most antitrichomonal activity was related to ethyl acetate extract with the least MIC of 500 µg/ml and mean of 1525 µg/ml, after 48 hrs incubation. And also, the lowest antitrichomonal activity was related to hydroalcoholic and methanolic extract with the least and mean MIC of 2000 µg/ml. The results of MLC and MIC tests were identical and this finding confirmed the trichomonacidal activity of the extracts. The drug suscebtibility testing showed that the T. vaginalis isoleates were susceptibale to metronidazole ranging from 3.1 to 6.2 µg/ml with a mean and standard deviation of 4.2 ± 1.5 µg/ml. Conclusion: This study showed that the extracts of P. lanceolata hav e a considerable activity on T. vaginalis parasite. Hence, further studies are needed to clear more details of antimicrobial properties of P. lanceolata compounds

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

Genetic Identification of Trichomonas vaginalis by Using the Actin Gene and Molecular Based Methods

Background: Trichomonas vaginalis is the agent of urogenital tract infection that causes human trichomoniasis with some serious health complications. More understanding about genetic features of the parasite can be helpful in the study of the pathogenesis, drug susceptibility and epidemiology of the infection. For this end,we conducted analysis of the actin gene of T. vaginalis by applying the PCR-SSCP(PCR-Single Stranded Conformational Polymorphism) and nucleotide sequencingmethod. Methods: Fifty T. vaginalis samples were collected from 950 women attending gynecology clinics in two cities of Iran, Hamadan and Tehran, from November 2010 to July 2011. After axenisation of isolates, all samples subjected to PCR-SSCP and nucleotide sequencing. Results: According to the SSCP banding patterns and nucleotide sequencing, seven sequence types were detected among the isolates. Alignment of the nucleotide sequences showed five polymorphic sites in the different strain types. Amino acid substitution was not observed in the nucleotide sequence translation of the all sequences. Conclusion: The actin gene analysis represents genetic diversity of T. vaginalis and it suggests that various strains can be responsible for clinically different trichomoniasis in infected individuals. It is expected that further studies will be conductedto increase our knowledge about relationship between the actin gene polymorphism and different biological behavior of the parasite

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