Perception of Ticks and Tick-Borne Diseases Worldwide.

In this comprehensive review study, we addressed the challenge posed by ticks and tick-borne diseases (TBDs) with growing incidence affecting human and animal health worldwide. Data and perspectives were collected from different countries and regions worldwide, including America, Europe, Africa, Asia, and Oceania. The results updated the current situation with ticks and TBD and how it is perceived by society with information bias and gaps. The study reinforces the importance of multidisciplinary and international collaborations to advance in the surveillance, communication and proposed future directions to address these challenges.

Predicting climate-driven distribution shifts in Hyalomma marginatum (Ixodidae).

Hyalomma marginatum is an important tick species which is the main vector of Crimean­Congo haemorrhagic fever and spotted fever. The species is predominantly distributed in parts of southern Europe, North Africa and West Asia. However, due to ongoing climate change and increasing reports of H. marginatum in central and northern Europe, the expansion of this range poses a potential future risk. In this study, an ecological niche modelling approach to model the current and future climatic suitability of H. marginatum was followed. Using high-resolution climatic variables from the Chelsa dataset and an updated list of locations for H. marginatum, ecological niche models were constructed under current environmental conditions using MaxEnt for both current conditions and future projections under the ssp370 and ssp585 scenarios. Models show that the climatically suitable region for H. marginatum matches the current distributional area in the Mediterranean basin and West Asia. When applied to future projections, the models suggest a considerable expansion of H. marginatum's range in the north in Europe as a result of rising temperatures. However, a decline in central Anatolia is also predicted, potentially due to the exacerbation of drought conditions in that region.

Molecular Detection of Tacheng Tick Virus-1 (TcTV-1) and Jingmen Tick Virus in Ticks Collected from Wildlife and Livestock in Turkey: First Indication of TcTV-1 Beyond China.

Background: Jingmen tick virus (JMTV) and Tacheng tick virus-1 (TcTV-1) are emerging tick-borne viruses that have been recently confirmed to be etiological agents of human disease in China. However, the ecology of JMTV and TcTV-1, especially their association with ticks in wildlife and livestock, remains largely unknown in Turkey. Materials and Methods: Eight hundred thirty-two tick specimens in 117 pools were collected in Turkey between 2020 and 2022 from wildlife (Miniopterus schreibersii and Rhinolophus hipposideros; n = 10, 1.2%; Testudo graeca; n = 50, 6%) and livestock (Ovis aries and Capra aegagrus hircus; n = 772, 92.7%). The specimens were individually screened for JMTV and TcTV-1 using nRT-PCR assays targeting the partial genes. Results: JMTV was detected in one Ixodes simplex pool and two Rhipicephalus bursa pools collected from central and Aegean provinces, respectively. TcTV-1 was identified in five Hyalomma aegyptium pools collected in Mediterranean provinces. No coinfection was detected in the tick pools. Maximum likelihood analysis of JMTV partial segment 1 sequences reveal that these sequences form a separate cluster together with viruses previously characterized in Turkey and the Balkan Peninsula. The phylogenetic analysis of the TcTV-1 nucleocapsid sequences indicates that they are closely related to viruses in ticks, sheep, cattle, and humans in China, but form a separate group among themselves. Conclusion: This study provides the first molecular evidence of TcTV-1 in Hy. aegyptium in Turkey. In addion, these findings indicate that JMTV and TcTV-1 extend ticks species and geographic distributions. Thus, multiregional surveillance in livestock and wildlife is needed to evaluate potential tick vectors and the human health impact of these viruses in Turkey.

Phylogenetic placement of Turkish populations of Ixodes ricinus and Ixodes inopinatus.

Studies on phylogeography and population structure of Ixodes ricinus have been carried out in Europe for decades, but the number of specimens from the Middle East included in these analyses is relatively small, despite the wide distribution of the species in this area. This study aimed to clarify the phylogenetic positions of I. ricinus from Turkey as well as to investigate the presence of Ixodes inopinatus in Anatolia. For this purpose, one mitochondrial (mt 16S rDNA) and one nuclear gene (defensin) were used to generate molecular data from I. ricinus samples, which were collected from 17 locations across the species' distributional range in Turkey. Bayesian inference was used to investigate phylogenetic relationships. Globally, the mt 16S rDNA lineages correspond to the lineages revealed by defensin; I. ricinus and I. inopinatus sequences clustered separately. However, a discordant genetic pattern was observed between the phylogenetic position of turkish I. ricinus revealed by nuclear versus mitochondrial genes. All Turkish haplotypes of mt 16SrDNA clustered with I. ricinus samples from Europe, which might be the result of extensive gene flow between populations of Europe and the Middle East. On the other hand, a sample from Thrace Region grouped within I. inopinatus clade. Thus, the occurrence of I. inopinatus in Turkey was demonstrated for the first time using molecular data. Moreover, four individuals were found to be heterozygous for the defensin. The potential evolutionary processes that underlie this observed discrepancy between the phylogenetic trees of two genes have been discussed.

A survey of hard ticks associated with cave dwelling mammals in Turkey.

Members of the subgenus Eschatocephalus Frauenfeld, 1853 are highly specialized cave-dwelling ectoparasites of bats. We conducted a comprehensive field-based survey on bat ticks in Turkey and provided information about the phylogenetic placement of collected species. Sampling was carried out at 26 caves from 18 provinces around Turkey between 2019 and 2021. Eighty-one tick specimens collected from the cave environment or on various cave roosting bats resulted in five species: Ixodes vespertilionis, Ixodes simplex, Ixodes ariadnae, Ixodes kaiseri, and Haemaphysalis erinacei. While I. simplex was the most frequently collected species with a rate of 56.2% mainly from bats (Miniopterus schreibersii), I. vespertilionis was the most prevalent species (65.4%) and found mainly on cave walls. The first record of I. ariadnae was provided for Turkey. Phylogenetic trees were built using mt 16S rDNA and COI markers. Our results demonstrated the presence of two distinct lineages of I. vespertilionis in Turkey; one lineage grouped with European isolates, whereas three sequences clustered separately. The phylogenetic pattern of I. simplex was consistent with previous results; this clade was clustered distantly to other bat tick species. The significance of the surprising records of H. erinacei and I. kaiseri in caves is also discussed.

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.

A cross-sectional screening by next-generation sequencing reveals Rickettsia, Coxiella, Francisella, Borrelia, Babesia, Theileria and Hemolivia species in ticks from Anatolia.

BACKGROUND: Ticks participate as arthropod vectors in the transmission of pathogenic microorganisms to humans. Several tick-borne infections have reemerged, along with newly described agents of unexplored pathogenicity. In an attempt to expand current information on tick-associated bacteria and protozoans, we performed a cross-sectional screening of ticks, using next-generation sequencing. Ticks seeking hosts and infesting domestic animals were collected in four provinces across the Aegean, Mediterranean and Central Anatolia regions of Turkey and analyzed by commonly used procedures and platforms. RESULTS: Two hundred and eighty ticks comprising 10 species were evaluated in 40 pools. Contigs from tick-associated microorganisms were detected in 22 (55%) questing and 4 feeding (10%) tick pools, with multiple microorganisms identified in 12 pools. Rickettsia 16S ribosomal RNA gene, gltA, sca1 and ompA sequences were present in 7 pools (17.5%), comprising feeding Haemaphysalis parva and questing/hunting Rhipicephalus bursa, Rhipicephalus sanguineus (sensu lato) and Hyalomma marginatum specimens. A near-complete genome and conjugative plasmid of a Rickettsia hoogstraalii strain could be characterized in questing Ha. parva. Coxiella-like endosymbionts were identified in pools of questing (12/40) as well as feeding (4/40) ticks of the genera Rhipicephalus, Haemaphysalis and Hyalomma. Francisella-like endosymbionts were also detected in 22.5% (9/40) of the pools that comprise hunting Hyalomma ticks in 8 pools. Coxiella-like and Francisella-like endosymbionts formed phylogenetically distinct clusters associated with their tick hosts. Borrelia turcica was characterized in 5% (2/40) of the pools, comprising hunting Hyalomma aegyptium ticks. Co-infection of Coxiella-like endosymbiont and Babesia was noted in a questing R. sanguineus (s.l.) specimen. Furthermore, protozoan 18S rRNA gene sequences were detected in 4 pools of questing/hunting ticks (10%) and identified as Babesia ovis, Hemolivia mauritanica, Babesia and Theileria spp. CONCLUSIONS: Our metagenomic approach enabled identification of diverse pathogenic and non-pathogenic microorganisms in questing and feeding ticks in Anatolia.

A metagenomic survey identifies Tamdy orthonairovirus as well as divergent phlebo-, rhabdo-, chu- and flavi-like viruses in Anatolia, Turkey.

We employed a direct metagenomic approach via next-generation sequencing for a cross-sectional investigation of viruses in 10 tick pools, collected from Aegean, Mediterranean and central Anatolian locations in Turkey. Sequences from all genome segments of Tamdy orthonairovirus (family Nairoviridae) were characterized in ticks collected from a Meriones tristrami. We further obtained near-complete L and partial S segments of several tick-associated phleboviruses (family Phenuiviridae), including Tacheng tick virus 2 and a novel virus, tentatively named as the tick phlebovirus Anatolia. Partial NS5-coding region of recently-described flavi-like virus (Tacheng tick virus 8) was further detected. Moreover, near-complete and polymerase-coding regions of arthropod-associated rhabdoviruses as well as sequences closely-related to the members of the newly-proposed virus family, the Chuviridae, were characterized. Despite origins of the viral sequences could not be fully elucidated, the findings suggest the circulation of diverse arthropod and tick-associated viruses in Anatolia. Occurrence and outcome of vertebrate exposure and probable health impact of these viruses require further investigation. We also report the initial detection of Tamdy orthonairovirus, an established human pathogen, which should be included in the diagnostic workup of infections with unknown etiology.

Distribution and phylogeny of Hyalomma ticks (Acari: Ixodidae) in Turkey.

The genus Hyalomma includes some of the most medically and veterinarily important tick species in the world. To clarify and identify the current distribution of the species of Hyalomma, field studies were conducted in 65 localities in Turkey and five localities in Cyprus. Additionally, using mitochondrial 12S and 16S ribosomal DNA, specimens of Hyalomma from Turkey, H. excavatum from Cyprus, H. marginatum from Spain and Italy were evaluated together with the available sequences obtained from Genbank. Morphological and molecular analyses demonstrated the presence of four species in Turkey: H. marginatum, H. excavatum, H. aegyptium and H. asiaticum. Hyalomma marginatum is the dominant species in the Central and Northern parts of Turkey, whereas H. excavatum distributes mostly in the Southern parts. Hyalomma asiaticum is restricted to the Southeastern Anatolia. However, some sympatric regions were observed for these species. Phylogenetic trees obtained with Maximum Likelihood method demonstrated five clades. Data supported previous conclusions, but placed H. asiaticum, H. scupense, H. dromedarii and H. aegyptium in different clades with high bootstrap values. Specimens of H. anatolicum group and H. marginatum complex are sister groups. Pairwise distance analyses of these groups showed 2.8 and 3% differences for 12S rDNA and 16S rDNA, respectively. Therefore, additional analyses with the samples from different locations using different markers need to evaluate the exact status of the species of these groups.

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.