Molecular prevalence and genotyping of Giardia duodenalis in cattle in Central Anatolia Region of Turkey.

The molecular prevalence and genotypes of Giardia duodenalis in cattle were investigated. A total of 450 fecal samples were collected from cattle in three provinces of Central Anatolia from August 2017 to July 2019. Genomic DNA was extracted from the fecal samples and used in molecular analysis carried out by nested PCR analyses of the ß-giardin (bg) gene of G. duodenalis. Positive samples were further analyzed by nested PCR at two gene loci (triosephosphate isomerase (tpi) and glutamate dehydrogenase (gdh)) for genotyping of G. duodenalis isolates. PCR analyses of the bg gene indicated that the overall prevalence of G. duodenalis was 30.2%. However, lower rates were determined with PCR analyses for gdh and tpi loci. The sequence analyses of the bg, gdh, and tpi genes revealed the presence of zoonotic assemblage A and livestock-specific assemblage E. Combined-sequence analyses revealed that assemblage E was the most common in the study area. Our study provides the first data on the wide prevalence of livestock-specific assemblages E in cattle in Turkey. The prevalence of assemblage A in cattle also reveals the importance of cattle for zoonotic transmission of giardiasis in Turkey.

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.

First Molecular Detection and Phylogenetic Analyses of Zoonotic Giardia intestinalis in Horses in Turkey.

The goal of our study was to investigate the molecular prevalence of Giardia intestinalis in naturally infected horses in Kayseri, Central Anatolia Region in Turkey, to determine the molecular characterization of the obtained isolates and to exhibit the potential role of horses in zoonotic transmission of G. intestinalis. Fecal samples were randomly collected from totally 150 horses with clinically healthy between March and June of 2018. After the genomic DNA extractions, 25 (16.6%) of the 150 fecal samples, were found positive for G. intestinalis by nested PCR analyses of ß-giardin gene. Phylogenetic analysis of the ß-giardin gene sequences of G. intestinalis showed that the sequences detected in this study belonged to assemblage A that is regarded as zoonotic. Our study is the first report on the presence of G. intestinalis in horses in Turkey. The findings of the present study indicate that future research studies are required to determine molecular epidemiology and geographical distribution of G. intestinalis infections in horses nationwide. In addition, this study also may be helpful to assess the zoonotic potential for public health of G. intestinalis infections.

Molecular detection and identification of Wolbachia endosymbiont in fleas (Insecta: Siphonaptera).

The aim of this study was to determine the presence and prevalence of Wolbachia bacteria in natural population of fleas (Insecta: Siphonaptera) in Turkey, and to exhibit the molecular characterization and the phylogenetic reconstruction at the positive isolates with other species in GenBank, based on 16S rDNA sequences. One hundred twenty-four flea samples belonging to the species Ctenocephalides canis, C. felis, and Pulex irritans were collected from animal shelters in Kayseri between January and August 2017. All flea species were individually screened for the presence of Wolbachia spp. by polymerase chain reaction (PCR) targeting the 16S ribosomal RNA gene. According to PCR analyses, Wolbachia spp. were found prevalent in C. canis and P. irritans fleas, while it was not detected in the C. felis species. Totally, 20 isolates were purified from agarose gel and sequenced with the same primers for molecular characterization and phylogenetic analyses. The sequence analyses revealed 17 polymorphic sites and 2 genetically different Wolbachia isolates, representing two different haplotypes in two flea species. The distribution patterns, molecular characterization, and phylogenetic status of Wolbachia spp. of fleas in Turkey are presented for the first time with this study. Understanding of the role of Wolbachia in vector biology may provide information for developing Wolbachia-based biological control tools.

Molecular Characterization of the Horn Fly Haematobia irritans Infesting Horses in Central Anatolia Region in Turkey.

This study reports intense horn fly infestations of horses raised in an important wetland ecosystem, Sultan Marshes in Central Anatolia, Turkey. In total, seven horses raised together were found to be harbored over 500 flies per site of each animal. Totally, 376 fly specimens were collected from the horses by using the nets and were subjected to the laboratory for species identification. All flies were morphologically identified as the adults of Haematobia irritans. Partial fragments of mitochondrial cytochrome oxidase I (mt-COI) gene from totally 50 isolates were amplified for sequence and phylogenetic analyses. The mt-COI sequence analyses revealed no polymorphism among the isolates and explored a unique haplotype for H. irritans. A mean haplotype and nucleotide diversities of 0.8571 and 0.00695 were determined, respectively, within the COI data set of H. irritans, and newly characterized haplotype from Turkey exhibited a mean intraspecific genetic difference of 1.0 to the all published sequences of the isolates from several countries. COI data set also revealed a mean interspecific genetic difference of 1.7 between H. irritans and Haematobia exigua.