Molecular Survey of Anaplasma capra in Goats in Van Province, Eastern Türkiye.

BACKGROUND: A newly discovered zoonotic infection carried by ixodid ticks, Anaplasma capra, affects a wide variety of hosts, including numerous mammals. A. capra most likely infects erythrocytes or endothelial cells in mammals. This study aimed to investigate the A. capra pathogen in goats in Türkiye's Van province. METHODS: A total of 200 goat blood samples were examined. Goat samples were subjected to partial amplification of the gltA gene fragment using a nested polymerase chain reaction. RESULTS: A. capra DNA was detected in 0.5% of goat blood samples. Phylogenetic analysis of a partial gltA gene fragment showed that the Eastern Türkiye isolate, closely grouped with A. capra isolates reported from wild and domestic ruminants in France, Türkiye, and Kyrgyzstan, formed a distinct clade. CONCLUSIONS: This is the first report of A. capra in goats in Van province, Eastern Türkiye.

Impact of nanopore-based metagenome sequencing on tick-borne virus detection.

Introduction: We evaluated metagenomic nanopore sequencing (NS) in field-collected ticks and compared findings from amplification-based assays. Methods: Forty tick pools collected in Anatolia, Turkey and screened by broad-range or nested polymerase chain reaction (PCR) for Crimean-Congo Hemorrhagic Fever Virus (CCHFV) and Jingmen tick virus (JMTV) were subjected to NS using a standard, cDNA-based metagenome approach. Results: Eleven viruses from seven genera/species were identified. Miviruses Bole tick virus 3 and Xinjiang mivirus 1 were detected in 82.5 and 2.5% of the pools, respectively. Tick phleboviruses were present in 60% of the pools, with four distinct viral variants. JMTV was identified in 60% of the pools, where only 22.5% were PCR-positive. CCHFV sequences characterized as Aigai virus were detected in 50%, where only 15% were detected by PCR. NS produced a statistically significant increase in detection of these viruses. No correlation of total virus, specific virus, or targeted segment read counts was observed between PCR-positive and PCR-negative samples. NS further enabled the initial description of Quaranjavirus sequences in ticks, where human and avian pathogenicity of particular isolates had been previously documented. Discussion: NS was observed to surpass broad-range and nested amplification in detection and to generate sufficient genome-wide data for investigating virus diversity. It can be employed for monitoring pathogens in tick vectors or human/animal clinical samples in hot-spot regions for examining zoonotic spillover.

Several Tick-Borne Pathogenic Viruses in Circulation in Anatolia, Turkey.

Introduction: We screened host-collected ticks for tick-borne viruses, including those recently documented as human pathogens. Methods: During 2020-2021, ticks removed form cattle, sheep, dogs, and cats in 11 provinces in 5 geographically distinct regions of Anatolia were identified, pooled, and screened using pan-nairovirus, pan-flavivirus and individual assays for Jingmen tick virus (JMTV), and Tacheng tick virus 1 and 2 (TcTV-1 and TcTV-2). Results: A total of 901 tick specimens, comprising 6 species were included. Rhipicephalus sanguineus complex was the most abundant species (44.1%), followed by Rhipicephalus bursa (38.3%), Haemaphysalis parva (7.2%), and others. The specimens were screened in 158 pools with 12 pools (7.6%) being positive. Crimean-Congo hemorrhagic fever virus (CCHFV) lineage Europe 2 (genotype VI) sequences were detected in R. bursa in five (3.2%) of the pools, with similar prevalences in central and Mediterranean Anatolian provinces. JMTV was identified in four R. bursa and one Rhipicephalus turanicus pools, collected from Mediterranean and southeastern Anatolia, with a CCHFV and JMTV coinfected R. bursa pool. The JMTV segment 1 sequences formed a separate cluster with those from Turkey and the Balkan peninsula in the maximum likelihood analysis. TcTV-2 was detected in two Dermacentor marginatus specimens (1.3%) collected in central Anatolia, with nucleocapsid sequences forming a phylogenetically segregated group among viruses from humans and ticks from China and Kazakhstan. Discussion: CCHFV Europe 2 was initially documented in ticks from central Anatolian locations, where related orthonairoviruses had been previously recorded. Ongoing activity and a wider distribution of JMTV and TcTV-2 were observed. These viruses should be screened as potential etiological agents in human infections associated with tick bites.

Genetic Analysis of Toxocara Spp. in Stray Cats and Dogs in Van Province, Eastern Turkey.

INTRODUCTION: Toxocara 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 METHODS: Adult 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. RESULTS: It 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. CONCLUSION: To 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.

Preliminary Studies on the Prevalence and Genotyping of Echinococcus Granulosus Infection in Stray Dogs in Van Province, Turkey.

INTRODUCTION: Echinococcus 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 METHODS: This 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. RESULTS: Taeniidae 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). CONCLUSION: Dogs in Van area are primarily infected with the livestock genotype of E. granulosus, which is thought to be a potential zoonotic threat to humans.

Generic amplification and next generation sequencing reveal Crimean-Congo hemorrhagic fever virus AP92-like strain and distinct tick phleboviruses in Anatolia, Turkey.

BACKGROUND: Ticks 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. RESULTS: A 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. CONCLUSIONS: Widespread 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.

[Cystic echinococcosis and cysticerci of Taenia hydatigena in cattle and sheep slaughtered in a Van Local Slaughterhouse]. / Van Belediye Mezbahasinda Kesilen Sigir ve Koyunlarda Taenia hydatigena Sistiserkozusu ve Kistik ekinokokkozis.

OBJECTIVE: The purpose of this study on cattle and sheep slaughtered in the Van Local Slaughterhouse, in which some metacestodes were found (Cysticercus tenuicollis, Cyst hydatid), was to determine the status and the prevalence of these parasites which are still widely seen in various regions of Turkey and to determine the distribution of internal organs. METHODS: Organ inspections were performed manually and visually on 525 sheep and 184 cattle which were bred in the Van region and slaughtered in the Van Local Slaughterhouse in the Van region. RESULTS: Five hundred twenty-five sheep and 184 cattle were examined for hydatidosis and the infection rates were found to be 44.4% and Cysticercus tenuicollis infection rate was 27.9%. CONCLUSION: This study indicated that, in slaughtered animals, meat is a common source of cysticerces and the importance cannot be underestimated.

[Cytochrome oxidase I gene sequences that cause hypodermosis cattle species by PCR-RFLP technique investigation]. / Sigirlarda Hypodermosise Sebep Olan Türlerin Sitokrom Oksidaz I Gen Dizilerinin PCR-RFLP Teknigi ile Arastirilmasi

OBJECTIVE: The importance for the health of cattle Hypodermosis (Nokra) species that cause disease by Polymerase Chain Reaction- Restriction Fragment Length Polymorphism (PCR-RFLP) method was investigated. METHODS: The molecular analysis of Hypoderma species specific sequences located on the Cytochrome C Oxidase I (COI) (mtDNA) gene by PCR-RFLP method was successfully carried out. RESULTS: DNAs were obtained using the extraction kit and Polymerase chain reaction (PCR) foe amplification of the mitochondrial DNA gene yielded for 688bp COI gene on agarose gel. Using the PCR amplification and HinfI, RsaI and TaqI restriction enzymes, mitochondrial DNA gene regions were amplified with the RFLP technique. CONCLUSION: The mitochondrial COI gene region of the genus Hypoderma for the molecular identification of potential answers questions, such as the level, which indicates that it may be very useful. In this study, in the city of Van, it is also important that the Hypoderma species be defined in terms of the first molecular study.