Comparison of resistant and susceptible strains of Trichomons vaginalis to metronidazole using PCR method

Metronidazole is drug of choice recommended by WHO for treatment of trichomoniasis, however, some reports claims drug resistance in Trichomonas vaginalis isolates recently. The objective of this study was to determine the minimum lethal concentration [MLC] of metronidazole in resistant and sensitive strains, as well as genetic patterns of these stains by PCR method. From February 2006 to March 2007, in a cross sectional study, clinical and wet mount examination of vaginal smear along with culture were performed on 683 women attending to public and private outpatient clinics in Hamadan. Trichomoniasis marked based on major clinical symptoms. Diagnosis confirmed using wet mount microscopically and culture in Diamond medium. A serial concentration of metronidazole was provided and all isolated Trichomonas strains [resistant and sensitive] tested by standard method. Finally, all sensitive and resistant strains examined by PCR technique. Only 15/683, [2.2%] of patients clinically diagnosed trichomonal vaginitis were positive for T. vaginalis by wet smear and culture. The minimum lethal concentration [MLC] for clinically sensitive isolates was 25 micro g/ml; however, this concentration for resistant isolates was 200 micro g/ml after 24 h and 100 micro g/ml after 50 h. The results of PCR examination of DNA from sensitive and resistant isolates had same pattern. The lanes appeared by two primers were 98 bp and 261 bp for both clinically sensitive and resistant strains. Resistance to metronidazole in T. vaginalis has not relation to genetic variations and might be related to some physiologic pathways of organism

[Prevalence of IL18-607A/C polymorphism in patients with visceral leishmaniasis]

Visceral leishmaniasis [VL] is a parasitic disease caused by a protozoan of Leishmania genus and in Iran by Leishmania infantum. The protective immune response against VL is cellular immunity through Th1 CD4+, which dominant chemokiens are IL12, IFN-alpha and IL18 and lead to Th1 response. Single nucleotide polymorphism [SNP] on IL-18 gene and its relation to IL18 levels in blood and IL18 function have been studied in many inflammatory diseases such as Behcect's disease and tuberculosis. According to the important role of IL-18 in immunity against visceral leishmaniasis, this study was conducted to demonstrate the prevalence of genotypes on-607A/C in promoter region of IL-18 gene. This descriptive and cross-sectional study was done on 91 pateints with confirmed VL, 105 healthy sero-negative controls and 78 seropositive controls during 1999-2009. Salting out method was used to extract DNA and ARMS-PCR was used to determine the genotype of-607A/C allele of individuals. Statistical analysis of genotypes was performed using Chi-Square test. According to the results,-607C/C was the dominant genotype among the groups [35.8%]. Distribution of genotypes among groups had not any significant difference. The lowest genotype among healthy sero-positive and patients were-607A/C and-607A/A, respectively. Statistical analysis of distribution of genotypes, did not reveal any significant difference among groups. The dominant genotypes of VL patients, healthy sero-negatives and healthy seropositives were-607C/C [38.5%],-607A/C [37.1%] and-607C/C [35.9%] respectively

Identification and genetic variation of fasciola species from Tabriz, north-western Iran

Fascioliasis is considered as the most important helminthic infection of cattle and sheep. Traditional approaches using morphlogical and biologic characters cannot cause a certainty in the accurate and precise identification and intra-specific differences of Fasciola spp. In this study, we identified Fasciola species using ITS-1 marker and described genetic variation of each species of the parasite in isolates from Tabriz slaughterhouse in West Azerbaijan Province, north-western Iran. Overall, 100 samples [50 from sheep and 50 from cattle] morphologically detected as Fasciola worms were studied for identification of Fasciola species by PCR-RFLP method and intra-species variation of the parasite using RAPD-PCR technique. A region of approximately 460bp in all samples was successfully amplified. There were no identifiable variations among the size of PCR products. Two and three fragments in samples correspond to F. hepatica and F. gigantica was seen, respectively, through PCR-RFLP method. No difference was seen in digestion pattern according to host [sheep or cattle]. Different types of each species of the parasite was observed using RAPD-PCR technique. We could have an estimate of frequency of F. hepatica and f. gigantic and different genotype of the parasite in isolates from one locality in north-western of Iran. By extension of such studies in future to other animal hosts [buffalo and goat] and including more regions to sampling, the reliability of the results and their application for control programs in zoonotic diseases will be increased

Association between en dothelial selectin [E-selectin] gene polymorphisms and E-selectin level with visceral leishmaniais, in an ARMS-PCR based study

In the visceral leishmaniasis [VL], parasites reside in reticuluendothelial system, mainly in macrophages. Endothelial Selectin [E-selectin] might play an important role in leukocyte-endothelium interactions and inflammatory cell recruitment. The aim of this study was determining E-selectin level and its polymorphism in three groups, patients, seropositive and healthy individuals. Serum soluble E-selectin levels as well as 2 polymorphisms of E-selectin [Ser 128 Arg and Leu 554 Phe] were measured in a cohort of patients with documented VL [n=64], a healthy control group [n=74] and a seropositive for VL but without any symptoms [n=81]. Circulation concentration of E-selectin levels was measured by ELIS. The amplification refractory mutation system [ARMS]-PCR procedure was used for detecting polymorphisms. The mean of E-selectin levels significantly differed between three groups [P<0.026], and were increased in patients in comparison with other groups. Difference was more considerable between two groups of patients and healthy ones [patients 92.8 ng/ml; healthy individuals 71.9 ng/ml]. Polymorphisms were associated with soluble E-selectin levels and altogether explained 14.4%, 7.2%, and 8.7% in patients, seropositive and seronegative healthy individuals, respectively. Distribution of polymorphisms of 128 Ser/Arg and 554 Leu/Phe among three groups was not different significantly; however, there was a considerable arrangement in distribution of Ser 128 Arg polymorphism and 128 Arg allele in healthy group was more than two fold of patients [55% against 20%]. The association between soluble E-selectin levels and visceral leishmaniasis suggests that this molecule might have significant role in the inflammatory process in VL. Moreover, frequency of 128 Arg allele in healthy group was higher than patients