Molecular characterization of cattle and sheep isolates of echinococcus granulosus, haplotype analysis, and determination of miRNA expression levels

Cystic Echinococcosis (CE) is a zoonotic helminth disease caused by the larval form of Echinococcus granulosus, which is common in humans and intermediate host animals worldwide. Detection of genetic differences in E. granulosus is important in terms of determining the pathogenicity of the disease, transmission route and control methods. One of the approaches trying to explain the mechanisms in the host-parasite relationship of E. granulosus is microRNAs. In this project, E. granulosus s.l. genotype and haplotype groups were determined in hydatid cyst isolates of cattle and sheep origin, and the expression levels of egr-miR-7, egr-miR-71 and egr-miR-96 miRNAs were compared in different cyst structures and between groups. The potential of target miRNAs to be used as a biomarker in the diagnosis of the disease was investigated. Genotypes of 81 out of 82 hydatid cyst isolates (41 cattle, 41 sheep) were determined by PCR and sequence analyzes for the mt-CO1 gene region of E. granulosus s.s. (G1 and G3), the genotype of an isolate of cattle origin was Echinococcus canadensis (G6/7). A total of 26 nucleotide polymorphisms and 29 haplotype groups were identified in the samples. miRNA expressions in germinal membranes of sterile cysts and germinal membrane and protoscoleces of fertile cysts were investigated by qRT-PCR and Real Time PCR analyses. E. canadensis (G6/7) sample was selected as control and miRNA expression levels in all haplotype groups were compared. It was determined that miRNAs were expressed at high levels in 79.31% of 29 haplotype groups and at low levels in 10.345%. In 10 fertile samples of sheep origin, egr-miR-7, egr-miR-71 and egr-miR-96 miRNAs were found to be 44, 168, and 351-fold higher in expression, respectively, in the germinal membrane compared to the protoscolex structure. High statistically significant differences in expression levels were detected between genotype and haplotype groups. In conclusion, it has been shown that egr-miR-7, egr-miR-71 and egr-miR-96 miRNAs, and especially egr-miR-96, may have the potential to be used as biomarkers in the diagnosis of active CE.