Molecular Prevalence and Phylogenetic Characterization of Enterocytozoon bieneusi in Sheep in the Van Region

Abstract

Objective: In this study, we aimed to determine the prevalence of Enterocytozoon bieneusi in healthy sheep in Van province using molecular techniques and to reveal genotypes of the detected isolates.

Methods: A total of 200 healthy appearance sheep comprise 38 male and 162 female, 32 preweaned, 38 postweaned lamb and 130 adult sheep from several farms in the Van region were included in the study between May and September 2021. Genomic DNA (gDNA) extractions were utilized on fecal samples collected from sheep by commercial kits, and E. bieneusi DNA was investigated by Nested polymerase chain reaction (PCR) amplifying ITS rRNA in the gDNA isolates. PCR products of the positive isolates were subjected to sequence analyze for genotyping and phylogenetic analyses of E. bieneusi.

Results: E. bieneusi DNA was determined in 16 out of 200 examined sheep fecal gDNA samples (8.0%) by Nested PCR. The highest E. bieneusi prevalence was determined in preweaned lambs with a rate of 18.8%. This was followed by postweaned lambs and adult sheep with a prevalence of 10.5% and 4.6%, respectively. The prevalence of the infection in males and females was 7.9% and 9.3%, respectively. All the ITS rRNA amplicons from 16 positive isolates were subjected to sequence analyses for genotyping and phylogenetic analyses. Sequence analyses revealed that all the isolates determined in sheep belonged to the BEB6 genotype and clustered in genogroup 2 of E. bieneusi with the BEB6 isolates from different hosts in several countries.

Conclusion: Molecular epidemiological data on the prevalence of E. bieneusi in sheep in Turkey were obtained with this study and the common genotype was determined as BEB6 in the research area. The obtained data contribute to the molecular epidemiology and diversity of E. bieneusi in sheep.