First report on the molecular prevalence of Enterocytozoon bieneusi in horses in Turkey: genotype distributions and zoonotic potential.

Horses might play an important role as reservoir hosts in the epidemiology of Enterocytozoon bieneusi, which is one of the most important zoonotic microsporidian pathogens, with a wide range of hosts. Nevertheless, limited information is available on the infection rates and genotypes of E. bieneusi in horses, and no data are available on the occurrence and molecular characteristics of E. bieneusi in horses in Turkey. We determined the prevalence of E. bieneusi among horses raised on farms from two provinces of Central Anatolia Region, by amplification of the partial small subunit ribosomal RNA gene using nested PCR. We identified the genotypes of E. bieneusi isolates by analyzing the ribosomal internal transcribed spacer (ITS) sequences. The overall prevalence of E. bieneusi was 18.7% (56/300), with no significant differences in infection rates among age groups or between genders of horses. Sequence analysis revealed eight genotypes: two known genotypes (ERUSS1, BEB6) and six novel genotypes (named ERUH2 to ERUH7). The genotype ERUSS1 was the most common and was found on all farms, age groups, and genders. Phylogenetic analysis clustered all the identified genotypes in ruminant-specific group 2. Our findings contribute to the molecular epidemiology of E. bieneusi.

Molecular Prevalence and Phylogenetic Characterization of Enterocytozoon bieneusi in Healthy Cattle

Objective: In this study, it was aimed to determine the molecular prevalence and genotypes of Enterocytozoon in healthy cattle. Methods: Fecal samples were collected from 50 cattle in Sivas between October 2017 and March 2018 and genomic DNA (gDNA) isolations were performed. gDNA isolates were processed by Nested PCR specifically amplifying ITS rRNA gene region to identify E. bieneusi. ITS rRNA region of E. bieneusi positive isolates were sequenced for genotyping and phylogenetic analyzes. Obtained sequences were assembled with appropriative genetic software, then phylogenetic relationships were revealed. Results: According to Nested PCR analyses, 29 (19.3%) out of totally examined samples were found positive for E. bieneusi. As a result of the sequence analyses, five distinct genotypes were determined. The most frequent genotype ERUSS1 and the other ERUSS2-4 genotypes were characterized as close to each other, which was reported for the first time in the world. Two isolates were determined in N genotype that was reported from cattle in Germany and were more different from the other genotypes. Phylogenetic analysis revealed that all the genotypes characterized in the study belonged to the genogroup 2. Conclusion: First molecular epidemiological data on E. bieneusi in cattle from Turkey were obtained with this study.

Molecular Characterization and Phylogenetic Analyses of Oestrus ovis Larvae Causing Human Naso-pharyngeal Myiasis Based on CO1 Barcode Sequences

Objective: The identification and molecular characterization of the bot fly larvae from an infected human with naso-pharyngeal myiasis in Turkey were aimed in this study. Methods: A total of 8 bot fly larvae from a 49-year-old woman with naso-pharyngeal infection in Adana province constituted the materials of this study. Morphological identification was performed on the larvae according to described keys. The barcode region of the CO1 gene from the genomic DNA extracts of the larvae was amplified and sequence analyses were utilized. Haplotype and genetic distance analyses were performed in CO1 sequences and a phylogenetic tree was built revealing phylogenetic relationships. Results: All bot fly larvae were identified as second stage larvae of Oestrus ovis in terms of morphologic characteristics. There was no polymorphism among the CO1 sequences of all isolates leading to detection of a single novel haplotype. The newly characterized haplotype in this study clustered with the O. ovis haplotypes from Bosnia and Herzegovina, Croatia, Brazil, and Iran in a monophyletic clade with an overall identity of 99.5%. Interspecific genetic differences among the subfamilies of Oestridae were in the range of 19.8% to 30.8%. Conclusion: This study has provided the first molecular characterization data on O. ovis larvae from an accidental human host in Turkey based on CO1 barcode sequences.