BackgroundTicks are involved with the transmission of several viruses with significant health impact. As incidences of tick-borne viral infections are rising, several novel and divergent tick- associated viruses have recently been documented to exist and circulate worldwide. This study was performed as a cross-sectional screening for all major tick-borne viruses in several regions in Turkey. Next generation sequencing (NGS) was employed for virus genome characterization. Ticks were collected at 43 locations in 14 provinces across the Aegean, Thrace, Mediterranean, Black Sea, central, southern and eastern regions of Anatolia during 2014–2016. Following morphological identification, ticks were pooled and analysed via generic nucleic acid amplification of the viruses belonging to the genera Flavivirus, Nairovirus and Phlebovirus of the families Flaviviridae and Bunyaviridae, followed by sequencing and NGS in selected specimens.ResultsA total of 814 specimens, comprising 13 tick species, were collected and evaluated in 187 pools. Nairovirus and phlebovirus assays were positive in 6 (3.2%) and 48 (25.6%) pools. All nairovirus sequences were closely-related to the Crimean-Congo hemorrhagic fever virus (CCHFV) strain AP92 and formed a phylogenetically distinct cluster among related strains. Major portions of the CCHFV genomic segments were obtained via NGS. Phlebovirus sequencing revealed several tick-associated virus clades, including previously-characterized Antigone, Lesvos, KarMa and Bole tick viruses, as well as a novel clade. A wider host range for tick-associated virus strains has been observed. NGS provided near-complete sequences of the L genomic segments of Antigone and KarMa clades, as well as Antigone partial S segment. Co- infections of CCHFV and KarMa or novel phlebovirus clades were detected in 2.1% of the specimens.ConclusionsWidespread circulation of various tick-associated phlebovirus clades were documented for the first time in Anatolia. Genomes of CCHFV AP92 strains were identified in previously unexplored locations. NGS provided the most detailed genomic characterization of the Antigone and KarMa viruses to date. The epidemiological and health-related consequences must be elucidated.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-2279-1) contains supplementary material, which is available to authorized users.

This study was planned to determine the prevalences of the Nosema spp. and the black queen-cell virus (BQCV) among honeybees (Apis mellifera) raised in the province of Van by PCR and to determine the molecular characteristics of the determined isolates. A total of 260 adult worker bees from 26 colonies at 5 apiary locations belonging to the province of Van in April and May 2015 were collected for this reason. Samples were examined microscopically. In the case of positivity, spore identification was done by multiplex PCR. Reverse transcription/PCR analysis (RT/PCR) was carried out for the BQCV analysis. At the end of the microscopic examination, Nosema spp. spores were detected in 8 out of 26 colonies (32.5%). The result of multiplex-PCR revealed Nosema ceranae positivity in all of the samples, but no Nosema apis was determined. As a result of the RT/PCR tests of the samples BQCV was detected in 23 (88.5%) of the total 26 colonies. This study is the first to investigate Nosema spp. and BQCV with the PCR technique in bees raised in the province of Van.

IntroductionEchinococcus granulosus is a zoonotic helminth of the Taeniidae family living in the small intestines of dogs. The hydatid cyst, which is the larval form of this parasite, is observed in sheep, goat, cattle, and many other organisms including humans. It causes a disease called cystic echinococcosis. Identification of strains of E. granulosus in dogs is critical in parasite control and eradication where possible. This study aims to determine the genotype of E. granulosus eggs and prevalence of this parasite in the faeces of dogs in the Van Province using the copro-PCR method.Material and MethodsThis study was conducted between 2015 and 2016 on the faeces obtained from 100 stray dogs from different parts of the Van Province. The coprological examination was conducted using the formalin-ether concentration method.ResultsTaeniidae eggs were found in 10 (10%) out of 100 faecal samples. E. granulosus was detected in 4 out of 10 of these (40%) infected samples. Sequence analysis of positive amplicons obtained from PCR showed that there were sheep strains (G1).ConclusionDogs in Van area are primarily infected with the livestock genotype of E. granulosus, which is thought to be a potential zoonotic threat to humans.

SummaryThis study aimed to determine the presence and prevalence of viral and parasitic infections causing high rates of colony loss in honey bee colonies in Van province, eastern Turkey. Twenty-six different apiaries were collected from five counties in Van province. These samples were tested by Reverse-Transcriptase PCR (RT-PCR) for acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), black queen cell virus (BQCV) and deformed wing virus (DWV). Selected positives were sequenced, phylogenetically analyzed and investigated in terms of Varroa. DWV and BQCV were identified in 69.23% (18/26) and 88.46% (23/26) of the bees respectively whereas ABPV and CBPV were not detected in the sampled apiaries. Results of the phylogenetic analysis of DWV and BQCV sequences showed 94-100% similarity to DWV and BQCV isolates obtained from Genbank. Prevalence of varroasis was 89% (23/26) in Van. The obtained samples were identified as Varroa destructor by morphological investigation. The study showed that viral and parasitic agents commonly infect honeybees in Van province, with high prevalence rates for BQCV and DWV. There was also a high degree of conservation of DWV and BQCV sequences distinct from DWV and BQCV isolates from other geographical regions. These findings, including current prevalence and phylogenetic analysis data for DWV, BQCV and varroazis in honeybees, are useful for future studies.

IntroductionToxocara canis and Toxocara cati are roundworms of dogs and cats. The purpose of this study was to investigate the infection caused by these ascarids in cats and dogs, using microscopic and molecular analysis methods.Material and MethodsAdult ascarids were gathered from the faeces of dogs and cats in Van province, in 2015–2016. Existing keys and PCR sequencing of the ITS-2 fragment were used to identify the morphological features of the parasite species.ResultsIt was observed that out of 20 adult ascarids, 17 and 3 were found to be Toxocara canis and Toxocara cati, respectively. The ITS-2 gene region was amplified by PCR to perform molecular analysis. Genotyping indicated that the dogs and cats were infected with T. canis and T. cati, respectively, and none had Toxascaris leonina.ConclusionTo the best of our knowledge, this is the first report on the molecular characteristics of adult ascaridoid nematodes from cats and dogs in Turkey. The molecular approaches established in this study enable molecular identification and genetic structure studies of the ascaridoids.

Background: Toxocara vitulorum is a involved in the Ascaridoidea family and is a large roundworm with a semi translucent, soft body surface and pinkish color. Female worms measure 8-30cm in length, male worms 6-25cm. The major hosts of T.vitulorum are buffalo (Bubalus bubalis) and cattle (Bos species) in the humid tropics of Asia, Africa and South America. The diagnosis of T. vitulorum infections is usually made by observing characteristic eggs in routine fecal examination. Serological methods are also used to diagnose Toxocariasis. However, in recent years, PCR, a new generation molecular diagnostic method, has been used. The genetic structure of T. vitulorum is little known compared with data available from other parasites. The present sutudy was designed to determine the T. vitulorum isolates by the genetic characterization of the mitochondrial cytochrome c oxidase subunit I (cox1) gene.Materials, Methods & Results: Adult worms were collected from the feces of two calves (East Anatolian Red) during visits to the clinic at the Department of Internal Medicine of Van Yuzuncu Yil University, Faculty of Veterinary Medicine. Worms were washed thoroughly in 0.85 % saline to remove any debris and fixed into 70 % ethanol. After repeated and thoroughly washing the specimens, total genomic DNA of parasite extraction was performed be employing DNA extraction reagent kit (Thermo, GeneJET Genomic DNA Purification Kit) according to manufacturer’s recommendations. After DNA amplification, a 446 bp fragment of cox1 of T. vitulorum were obtained in all three isolates. All generated sequences were registered in GenBank database with accession numbers including MG905159, MG911729 and MG911730. The cox1 of T. vitulorum examined differed from another two isolates extracted from Germany beef cattle (KY313642.1) and Sri Lanka buffalo calf (FJ664617.1) at NCBI database. The MEGA 7 software was employed to calculate intra-species distance and similarity. The intra-species distance rate and similarity among the isolates were 0.005 and 99.995%, respectively. The cox 1 sequence of T. vitulorum did not differ from an isolate from Germany, but differed more from isolate from Sri Lanka. The phylogenetic tree that was constructed using the Neighbor-Joining (NJ) method. Bootstrap support (Bp) for ML trees was calculating 1000 bootstrap replicates. This results indicate that both the different species of Toxocara are host-specific and each member of the genus Toxocara spp. has a different about the molecular sequences. We used the phylogenies from the Maximum Parsimony (MP) method to construct another phylogenetic tree based on the cox1 (mtDNA) gene. The results again display that the cattle-calves (East Anatolian Red) isolates from Turkey homology with that obtained from the Germany beef cattle (accession no. KY313642.1).Discussion: The genetic analysis of parasites is a crucial factor in terms of determining epidemiology and the control parasitic diseases of humans and animals. Toxocara vitulorum is the most common gastrointestinal helmints infecting ruminants particularly in tropical regions. Phylogenetic analysis revealed that T. vitulorum is 100% homology with related to sequence of T. vitulorum from Germany. The characterization of cox1 region can provides a foundation for accurate identification of some helminth species using PCR. Even though the small sample size,  the obtained results might provide useful information for further phylogenetic studies on the family Ascaridae.

Background: Fasciolosis is an important food borne zoonotic disease caused by Fasciola trematode parasites. There are two types of  Fasciola spp. namely F. hepatica and F. gigantica, widely distributed across the globe, affecting both human and animal hosts. In endemic regions, it is possible to base the diagnosis of fasciolosis on clinical signs and the season, however, it could be more useful to support these data with fecal examination and various hematologic and serological tests. The aim of this study was to determine the prevalence of fasciolosis in cattle in Van province by copro-ELISA technique.Materials, Methods & Results: Fecal samples from 140 cattle were technically collected and examined by sedimentationzinc sulphate flotation technique. Modified McMaster sedimentation technique was applied to the egg positive samples to determine the EPG values. Fasciola hepatica coproantigens in samples were investigated by ELISA. The coprological and antigen ELISA prevalence of fasciolosis were determined as 5.07% and 30.7%, respectively, which shows the significant difference between these methods in examining the rate of infection. The highest prevalence of fasciolosis infection was observed in 1-2 age groups (41.9%), and this prevalence was followed by 3-5 (31.2%) and ≤6 age group (5%). The differences between age groups were found significant (P < 0.05). The prevalence in female and male cattle was found as 30.1% and 35.3% This difference was not found statistically significant (P > 0.05). The highest prevalence was observed in Brown Swiss with the ratio of 40% and this was followed by 31% in Crossbreed and 22.6% in Rubia Gallega. The differences among breeds were not statistically significant (P > 0.05).Discussion: Fasciola hepatica is the most common species of liver flukes, and its pathogenicity leads to significant impact on the economy of the livestock industry. The economic losses consist of costs of anthelmintics, drenches, labor, liver condemnation at meat inspection; and losses in production due to mortality, reduction in meat, milk and reduction in growth rate, fertility and decreased feed intake, conversion and lower resistance to other disease.To diagnose fasciolosis, eggs can only be detected in feces after the tenth or twelfth week of infection once the parasites have matured. It is reported that routine microscopic methods used before this stage do not provide sufficient information about the current infection status. Therefore, serological tests have been introduced for the early diagnosis of the disease. Among these tests, the ELISA test based on detecting antigens has become the most commonly used test. It is known that the probability of ELISA to cross-react with parasites that carry similar immunogenic features and the similarities between antibodies generated in previous infections and new infections pose a challenge to making the definitive diagnosis. Therefore, it is reported that to predict the parasitic potential of the host and the success of treatment beforehand, the presence of Fasciola spp. antigens can be investigated in serum instead of antibodies. In conclusion, this study has established prevalence of fasciolosis in cattle raised in Van province using the copro-ELISA technique for the first time. It has been concluded that copro-ELISA could serve as a useful technique for herd diagnosis of fasciolosis in cattle in addition to fecal examinations particularly with respect to fasciolosis.

This study indicated that, in slaughtered animals, meat is a common source of cysticerces and the importance cannot be underestimated.