ABSTRACT
Hydatidosis is one of the most important zoonotic parasitic diseases caused by the larval stage of Echinococcus granulosus which causes great health and economic losses. The aim of this study was to use the sequencing method to evaluate genotypes of E. granulosus isolated from humans and bovines using mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The samples were taken in the East Azerbaijan Province, Northwest Iran. Overall, 26 hydatid cyst samples (10 human and 16 cattle isolates) were collected. DNA extraction was taken from the protoscoleces of human and germinal layer of bovine samples. PCR was performed using the mitochondrial cytochrome c oxidase subunit 1(cox1) gene, and then it was sequenced. Sequences were analyzed for identification of their genotypes. All 16 bovine isolates were recognized as G1 genotypes (sheep strain) and G1B subtypes. Out of ten human host samples, seven isolates were G1B subtypes, and three samples were identified as G3 genotypes. The results of this study showed that G1 and especially G1B are the predominant genotype and subtype in humans and cattle in Northwest Iran.
ABSTRACT
Hydatidosis is one of the most important zoonotic parasitic diseases caused by the larval stage of Echinococcus granulosus which causes great health and economic losses. The aim of this study was to use the sequencing method to evaluate genotypes of E. granulosus isolated from humans and bovines using mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The samples were taken in the East Azerbaijan Province, Northwest Iran. Overall, 26 hydatid cyst samples (10 human and 16 cattle isolates) were collected. DNA extraction was taken from the protoscoleces of human and germinal layer of bovine samples. PCR was performed using the mitochondrial cytochrome c oxidase subunit 1(cox1) gene, and then it was sequenced. Sequences were analyzed for identification of their genotypes. All 16 bovine isolates were recognized as G1 genotypes (sheep strain) and G1B subtypes. Out of ten human host samples, seven isolates were G1B subtypes, and three samples were identified as G3 genotypes. The results of this study showed that G1 and especially G1B are the predominant genotype and subtype in humans and cattle in Northwest Iran.
ABSTRACT
BACKGROUND: In this study, we used both ITS1 and ITS2 for molecular identification of Fasciola species. METHODS: The region between 18S and 28S of ribosomal DNA was used in PCR-RFLP method for molecular identification of Fasciola species. Ninety trematodes of Fasciola were collected during abattoir inspection from livers of naturally infected sheep and cattle from Khorasan, East Azerbaijan, and Fars provinces in Iran. After DNA extraction, PCR was performed to amplify region ITS1, 5.8S rDNA, ITS2. To select a suitable restriction enzyme, we sequenced and analyzed the PCR products of F. hepatica and F. gigantica samples from sheep and cattle. Tsp509I fast digest restriction enzyme was selected for RFLP method that caused the separation specifically of Fasciola species. RESULTS: The fragment approximately 1000bp in all of the Fasciola samples was amplified and then digested with the Tsp509I restriction endonuclease. Seventy F. hepatica and 20 F. gigantica were identified of total 90 Fasciola isolates. CONCLUSION: The new PCR-RFLP assay using Tsp509I restriction enzyme provides a simple, practical, fast, low cost, and reliable method for identification and differentiation of Fasciola isolates.
