A molecular phylogenetic investigation of tick species in Eastern and Southeastern Anatolia.

Understanding the local tick species composition is crucial for overcoming the diseases they transmit. A comprehensive survey integrating molecular identification was conducted in the eastern and southeastern parts of Turkey, where tick surveys have previously been neglected. A total of 596 specimens belonging to four tick genera were collected from 27 localities in Turkey during the summers of 2019 and 2020. Seventy-seven representative individuals were chosen for molecular analysis. Nine distinct species, Rhipicephalus bursa, Rhipicephalus turanicus, Rhipicephalus rossicus, Hyalomma asiaticum, Hyalomma excavatum, Hyalomma marginatum, Hyalomma aegyptium, Haemaphysalis sulcata, and Dermacentor marginatus were identified. The presence of R. rossicus was demonstrated for the first time in Turkey. Two lineages of R. turanicus were identified, and representatives of both lineages were recorded. Our Hyalomma phylogenetic tree was consistent with previous findings from Turkey; however, new sympatric areas for Hy. marginatum and Hy. excavatum and Hy. marginatum and Hy. asiaticum were recorded. Two haplotypes (Haemaphysalis sp. and Dermacentor sp.) could not be identified using morphological and molecular methods. In addition to making a valuable contribution to the molecular database of ticks in the Middle East, this study will also stimulate comparative studies on the genetic structure, ecology, and vector competence of different populations of these species in Turkey as well as in other parts of the world.

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.

New molecular data shed light on the global phylogeny and species limits of the Rhipicephalus sanguineus complex.

The Rhipicephalus sanguineus complex is a group of closely related tick species distributed all around the world. In this study, using mitochondrial 16S ribosomal DNA, new specimens of R sanguineus sensu lato from Turkey and Rhipicephalus camicasi from Kenya, were evaluated together with available sequences of this complex in GenBank. Our objectives were to delimit the complex, re-evaluate its global phylogeny and develop a reconstruction of its biogeographic history. Given Turkey's geographical location and its neighboring status within Africa, Asia and Europe, molecular information of R. sanguineus s.l. species from this region could have important implications both on a regional and global scale. Phylogenetic trees obtained with three methods (Bayesian, Maximum Likelihood and Maximum Parsimony) were highly similar and consensus trees gave the same branching patterns and similar node support values. A total of four different clades with up to 9 Operational Taxonomic Units formed strong monophyletic groups. Biogeographic reconstructions demonstrated the importance of populations in Middle East (Turkey) in the spread of the group from Europe to Africa and Asia. Data supported previous conclusions on the existence of two species of R. sanguineus s.l. in South America and the strong molecular similarity between R. camicasi and the so-called tropical lineage of R. sanguineus s.l. These results point to the need of a re-evaluation of most specimens designated as R. sanguineus s.l. in East Europe, Middle East, Africa and Asia after an adequate re-description of this taxon.

Species distribution and detection of Crimean Congo Hemorrhagic Fever Virus (CCHFV) in field-collected ticks in Ankara Province, Central Anatolia, Turkey.

Ticks may act as vectors for a number of infectious diseases including Crimean Congo Hemorrhagic Fever (CCHF). The causative agent is Crimean Congo Hemorrhagic Fever Virus (CCHFV), a member of Bunyaviridae, causing extensive ecchymosis, visceral bleeding and hepatic dysfunction with a high fatality rate in the affected individuals. CCHF was initially recognized in Turkey in 2002 and the current number of reported cases exceeds 4,400. This study was conducted to confirm the presence of tick species established as potential CCHFV vectors and investigate CCHFV activity in ticks at Ankara province, Turkey's second most-densely populated province, where CCHF cases were demonstrated. A total of 1,196 adult ticks, collected from various animals and vegetation in 12 sites located in 5 counties of Ankara during April-July 2010 were identified to species level. Twenty-two tick pools from county K2 were also evaluated for the presence of CCHFV RNA via a one-step real-time RT-PCR assay and reactive results were further confirmed by an in house nested RT-PCR assay. Nine tick species were identified: Rhipicephalus bursa (44.9%), R. sanguineus (18.9%), R. turanicus (18.1%), Haemaphysalis parva (8.3%), Hyalomma marginatum marginatum (5.4%), H. aegyptium (1.4%), H. anatolicum excavatum (1.3%), Hae. punctata (0.3%) and Dermacentor marginatus (0.2%). A total of five tick pools (22.7%) were reactive in real-time and nested RT-PCR assays. The pools included R. bursa, H. m. marginatum and Hae. parva ticks, collected from mammal hosts from two villages in one county. This is the first documentation of CCHFV activity in ticks from Ankara province, which indicates requirement for detailed surveillance to predict high risk zones in the region.

Genetic and phenotypic variation of the malaria vector Anopheles atroparvus in southern Europe.

BACKGROUND: There is a growing concern that global climate change will affect the potential for pathogen transmission by insect species that are vectors of human diseases. One of these species is the former European malaria vector, Anopheles atroparvus. Levels of population differentiation of An. atroparvus from southern Europe were characterized as a first attempt to elucidate patterns of population structure of this former malaria vector. Results are discussed in light of a hypothetical situation of re-establishment of malaria transmission. METHODS: Genetic and phenotypic variation was analysed in nine mosquito samples collected from five European countries, using eight microsatellite loci and geometric morphometrics on 21 wing landmarks. RESULTS: Levels of genetic diversity were comparable to those reported for tropical malaria vectors. Low levels of genetic (0.004

Seroprevalence of West Nile virus and tick-borne encephalitis virus in southeastern Turkey: first evidence for tick-borne encephalitis virus infections.

West Nile virus (WNV) and tick-borne encephalitis virus (TBEV) are among the medically important Flaviviruses that cause significant morbidity and mortality in humans. In this study, seroprevalence of WNV and TBEV in sera from two state medical hospitals from the southeastern part of Turkey was investigated. One hundred eighty-one serum samples were evaluated for WNV immunoglobulin G (IgG) by an indirect immunofluorescence test (IIFT) and for IgG antibodies against TBEV by a commercial enzyme-linked immunosorbent assay (ELISA) kit with enhanced sensitivity and specificity. Sera positive for WNV IgG were further analyzed by plaque reduction neutralization assay (PRNA). TBEV IgM was also investigated by ELISA in all seroreactive samples. Of 181 sera, 29 (16%) were positive for WNV IgG by IIFT and 17 of 179 (9.5%) were confirmed by PRNA. Nineteen of 181 (10.5%) sera were detected to have TBEV IgG. Mean titer of TBEV IgG was 43.0 RU/mL (median, 33.9 RU/mL; cutoff: 20 RU/mL). Four samples with WNV IgG antibodies were also positive for TBEV IgG antibodies. TBEV IgM was detected in 9 of 39 (23%) of all seroreactive sera, where IgM positivity were accompanied by IgG for 6 samples. These results suggest the presence of possible human WNV and TBEV infections in southeastern Turkey where vector activity have previously been detected.

Distribution of sand fly (Diptera: Psychodidae) species and efficiency of capturing methods in Sanliurfa province, Turkey.

The population dynamics of sand flies (Diptera: Psychodidae) were studied in Sanliurfa province in southeastern Turkey, in the country's largest focus of typical anthroponotic cutaneous leishmaniasis, during 2000-2002. Sand flies were collected at nine different sampling stations, located throughout the city, representing a cross section of urban and rural habitats. In total, 29,771 sand flies were collected, 45.35% of which were Phlebotomus papatasi Scopoli. In this study, the overall sand fly species diversity, relative abundance of each species, biodiversity, and similarity indices among sampling stations and efficiency of trapping methods were evaluated. Among the sampling stations, Sanliurfa city center and Suruç were shown to have the highest number of sand fly species; Harran-Akcakale and Hilvan habitats produced the largest number of individuals. The greatest similarity rates (80%) in terms of sand fly species were observed between Hilvan and city center, Harran-Akcakale and city center, Harran-Akcakale and Yenice, and Siverek and Viransehir. The lowest similarity rate (16%) was observed between Bozova-Birecik and city center. Comparison of biodiversity and similarity indices between the various sampling stations reveals the distribution of the suspected vector species and provides basic knowledge required to develop logical and effective control strategies. Among the trapping methods used, light traps showed the highest capture efficiency, above aspirators and sticky papers. It was concluded that light traps alone were sufficient to determine the sand fly fauna of the study area. It is recommended that the spatial and temporal dynamics of sand fly populations be monitored throughout the southeastern Anatolia Project (GAP) construction period, considering the potential impact the project may have on mean temperature, humidity, and human population movements.

West Nile virus studies in the Sanliurfa Province of Turkey.

We attempted to isolate West Nile virus from mosquitoes collected in the field for the first time in Turkey. A total of 6,457 mosquito specimens from Culex pipiens Linnaeus, Ochlerotatus caspius (Pallas) and Aedes spp. species were included in this study. Culex pipiens samples made up 56% of the total species, O. caspius 24% and Aedes spp 20%. There were no positive results after studying mosquito samples using Real-time PCR, VecTest, and Vero cell culture. In serological tests of 181 human serum samples, 29 (16%) were found to be West Nile positive. On the basis of these results, we intend to collect more mosquito samples especially from those areas from which positive serum samples were obtained.

[West Nile virus and its vectors]. / Bati Nil virusu ve vektörleri.

There are more than five hundred known arthropod-borne viruses (arboviruses) all around the world and approximately hundred of them may cause disease in humans. During the past 20 years there has been a dramatic resurgence or emergence of epidemic arboviral diseases affecting both humans and domestic animals. Many factors play important roles in the emergence of arboviral diseases like Yellow Fever, Dengue, West Nile encephalitis, and of other diseases such as malaria and leishmaniasis in countries where they have not been previously encountered and in the increase in incidences where they have been under control. Some of these are demographic factors such as global population increase and uncontrolled urbanization; social changes such as modern transportation, human encroachment on natural disease hotspots; changes in agricultural activities such as the use of new irrigation techniques; deforestation; genetic changes in the pathogens; preventive measures and probably global climate changes. Mosquitoes are among the most important vectors carrying viruses belonging to Alphavirus, Flavivirus, Bunyavirus and Phlebovirus genera. All of the above factors have contributed to the increase in mosquito populations and closer contact between humans and mosquito vectors. West Nile virus notable after the epidemic of 1996 in Romania in Europe is one of the latest examples indicating that viruses can jump continents and produce epidemics. In this review article, the distribution of West Nile virus and its principal vectors and also its importance by means of public health, have been discussed.