Bidirectional tick transport by migratory birds of the African-Western Palearctic flyway over Turkish Thrace: observation of the current situation and future projection.

This study was carried out at a vital stopover site of migrating birds in the Turkish Thrace, European part of Turkey, on the Mediterranean/Black Sea Flyway. Ticks were collected from the birds captured in the four migration periods, i.e., autumn 2020, spring 2021, autumn 2021, and spring 2022, and identified morphologically. Throughout the study, 10,651 birds from 77 species were examined, and 671 belonging to 34 species were found infested. The infestation prevalence in total birds and the mean number of ticks per infested bird were 6.3% and 3.8 (range: 1-142), respectively. A total of 2573 ticks were collected with the following species distribution and numbers: Ixodes spp. 70 larvae, I. frontalis 1829 larvae, 337 nymphs, and 30 adults, I. acuminatus 16 nymphs and 42 adults, I. ricinus 39 larvae, 141 nymphs, and one adult, Hyalomma spp. seven larvae and 60 nymphs, and Haemaphysalis sp. one larva. Prevalence, intensity, and species distribution of the ticks in birds varied depending on the month, season, year, and species-specific migration phenology of the birds. The results show that precise determination of the tick-borne risk associated with migratory birds for a particular region necessarily requires long-term and comprehensive studies and indicates that anthropogenic climate change and habitat degradation can significantly differentiate the risk by influencing the migration phenology in birds and by making new regions suitable for the establishment of different ticks.

Detection of the Crimean-Congo Hemorrhagic Fever Virus Genome in Questing Ixodes spp. and Haemaphysalis spp. in the Periurban Forestry Areas of Istanbul: Has a New Biorisk Emerged?

Background: Istanbul is one of the world's most densely populated metropolitan cities, with various geographical areas that possess distinct characteristics. These areas have different climates, vegetation, and host populations that can support the survival of diverse tick species. Turkey is a region with a high risk of Crimean-Congo hemorrhagic fever (CCHF) disease, making it crucial to screen for this risk, especially in densely populated regions such as Istanbul and its surrounding areas. However, the presence of potential vectors for CCHF virus (CCHFV) in these areas has not been studied in the past 12 years. Materials and Methods: In this study, a total of 676 ticks were collected from 6 centers using the flagging and dragging method. Ticks were identified as Ixodes ricinus (7.85%), Ixodes spp. immature form (73.22%), Haemaphysalis parva (0.89%), Haemaphysalis inermis (0.15%), and Haemaphysalis spp. immature form (17.89%). Pooled samples were screened for the CCHFV genome (S segment) by RT-nested PCR. Results: A total of 14.28% of the samples were found as positive. Phylogenetic analysis revealed that all the CCHFV sequences obtained from the positive samples were clustered in clade V: Europe/Turkey genogroup. Conclusion: This study suggests that ixodes spp. and Haemaphysalis spp. may have the potential to pose a biorisk for Crimean-Congo hemorrhagic fever.

The Investigation of Giardiasis (Foodborne and Waterborne Diseases) in Buffaloes in Van Region, Türkiye: First Molecular Report of Giardia duodenalis Assemblage B from Buffaloes

Giardia duodenalis (G. duodenalis) is an important zoonotic protozoan agent that causes foodborne and waterborne diarrhea in humans and other mammals. Molecular-based tests are critical in diagnosing giardiasis in humans and animals, identifying species, understanding the zoonotic potential and transmission routes, and evaluating taxonomy. Therefore, this study aimed to investigate the molecular characterization of G. duodenalis in buffaloes in the Van region in Türkiye. Buffaloes are a species that has been poorly studied in this regard. For this purpose, 100 fecal samples were collected from buffaloes in the Van region. The DNA extraction was performed using the GeneMATRIX STOOL DNA Purification Kit from stool samples. The nested PCR test was performed with the appropriate primers from the obtained DNA samples. The obtained bands suitable for sequencing were sent for sequence analysis, and the sequence results were aligned bidirectionally and compared with the database of GenBank by BLAST. As a result of the study, an 11% positivity rate for G. duodenalis was found in buffaloes, and assemblage E and assemblage B were isolated. To our knowledge, assemblage B in buffaloes was reported for the first time in this study. As a result, it was concluded that buffaloes are an important reservoir for waterborne and foodborne giardiasis.

Prevalence and genotypes of Giardia duodenalis in shelter dogs of southeastern Türkiye.

Giardia duodenalis is a protozoan parasite found in humans and several mammals. This parasite spreads worldwide and is generally recognized as a zoonotic agent being reported to be one of the most common causes of diarrhea in humans and animals. In this study, it was aimed to determine the prevalence and genotypes of G. duodenalis in shelter dogs in Diyarbakir province being located in the southeastern Anatolia region of Türkiye. Native-Lugol method and nested polymerase chain reaction analyses of 100 fecal samples showed a prevalence of 3.00 and 4.00%, respectively. The prevalence was higher in females and in those younger than 1 year. Sequence analysis revealed the presence of zoonotic assemblage B, assemblage D and assemblage E. The detection of zoonotic assemblage B in this study suggests that dogs may be a reservoir for human giardiasis. Further molecular research is needed to determine the genotype diversity of Giardia as well as its possible role in the transmission of this parasite to humans.

Predation of ant species Lasius alienus on tick eggs: impacts of egg wax coating and tick species.

Several animal species, including ants, have been reported to be capable of predation on ticks. However, determining factors in most interactions between ticks and predators have not yet been fully deciphered. We hypothesized that the ant species Lasius alienus, which is unknown whether it has any impact on ticks, may exhibit predation on the eggs of tick species Hyalomma marginatum, H. excavatum, and Rhipicephalus bursa, and that the tick egg wax can be the main determinant in possible predation. In the study, 6300 tick eggs with the natural wax coating (waxed/untreated) and 2700 dewaxed tick eggs, the wax of which was removed in the laboratory, were repeatedly presented to the foraging workers belonging to three different ant nests in their natural habitat. Depending on the tick species and trials, the rate of the eggs carried by the ants ranged from 12.8 to 52.1% in the waxed and from 59.8 to 78.4% in the dewaxed eggs. It was observed that the dewaxing process both increased the interest of the ants in the eggs and resulted in a reduction in the variation associated with tick species. This study showed that L. alienus has a predatory effect on tick eggs, the severity of this impact is closely associated with the tick species, the tick-associated difference is caused by the species-specific property of the egg wax, and the variety in the protective effects of the wax seems to be an evolutional result of the biological and ecological adaptation process of the species.

Monthly infestation characteristics of ticks on cattle in Thrace, a Crimean Congo hemorrhagic fever-endemic area of Turkey.

The first etiologically confirmed cases of Crimean-Congo hemorrhagic fever (CCHF) in humans were detected in Turkey in 2002. Since then, thousands of cases have been reported from different parts of the country. Hyalomma (Hy.) marginatum is considered the main vector tick of CCHFe in Turkey, and the primary infection route for humans is known to be the tick bite. This study was carried out between January 2013 and December 2014 in Thrace, Turkey, to determine monthly prevalence and intensity of tick infestation in cattle and, ultimately, to predict the related risk of human exposure to ticks and tick-borne diseases. During the study, 1,701 cattle in 24 villages were screened for ticks; 24,012 adult ticks, 1,887 nymphs, and 766 larvae were encountered on 1,228 of these cattle. On the 1,318 cattle that routinely grazed in the daytime, Hy. marginatum was the most predominant species, with an infestation prevalence of 73.6% and an average tick intensity of 16.1 on the infested cattle. In order of prevalence (%) in the grazing cattle, Hy. marginatum was followed by Rhipicephalus (R.) bursa (32.7%), R. turanicus (%29.5), Ixodes (I.) ricinus (15.1%), Haemaphysalis (Ha.) parva (10.4%), Ha. inermis (10%), Ha. punctata (6.5%), and Dermacentor (D.) marginatus (2.3%). The obtained data were discussed in terms of the infestation characteristics of the observed tick species, the drivers that may affect these characteristics, and the features of possible relationship between Hy. marginatum infestation in the cattle and CCHF cases among humans in the area.

Monthly dynamics of the cold-adapted one-host biological north form of Hyalomma scupense under the influence of the warm summer subtype of the Mediterranean climate in Turkey.

The one-host biological north form / ecotype of Hyalomma scupense Schulze, 1919 (Acari, Ixodidae) is reported for the first time in Turkey herein. Following the first detection of the tick, a longitudinal field study was carried out to fill gaps in the data concerning its biological features. This study also aimed to determine how the monthly activation dynamics of this relatively cold climate-adapted species is characterized under the influence of the warm summer subtype of the Mediterranean climate. During this study, which was carried out on a monthly basis in 2014, H. scupense was found on cattle from 5 out of 18 villages screened in Thrace (the European part of Turkey). The field study revealed that i) this north ecotype of H. scupense exhibits winter one-host behavior beginning in October (with larval stages) and ending in April (with engorged adults); ii) engorged females detach from the cattle, drop on the floors of barns during night and accumulate on piled bedding contaminated with slurry manure; iii) in the late spring, engorged females lay eggs, and larvae hatch in the same area; and iv) larvae become active in autumn as the weather grows cooler. The results indicated that although one-host H. scupense is known to be distinctly adapted to cold conditions, it can also be effectively established in relatively temperate regions and complete its life cycle with some modifications in the timing of its monthly activation dynamics.

Phylogenetic and phylogeographic analysis of Myrmeleotettix maculatus (Orthoptera: Acrididae: Gomphocerinae) species group in Anatolia.

Six Anatolian and one European populations of the Myrmeleotettix maculatus species group, which contains M. maculatus and M. ethicus species, have been studied by using molecular genetics methods with mitochondrial COI gene. Myrmeleotettix ethicus is an Anatolian endemic species with local distribution whereas M. maculatus is distributed in western Palearctic. The phylogenetic analysis (ML and BI analyses) of the M. maculatus species group in Anatolia reveals that it consistently recovered two well-supported main clades and four different lineages. Molecular time estimates suggest that the diversification of the M. maculatus species group took place between the Late Tortonian (around 8-9 My) and the Middle of Pliocene-Pleistocene (around 4.3 My-present) periods and the current distribution of the genetic diversity has been affected by the uplifting of the Central Anatolian plateau, the termination of the Messinian salinity crisis, and the Quaternary climatic changes.

High prevalence and different genotypes of Crimean-Congo hemorrhagic fever virus genome in questing unfed adult Hyalomma marginatum in Thrace, Turkey.

Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral disease in the old continents, in many countries south of the 50 °North Parallel. The agent is known to be the most prevalent and major cause of severe and fatal human hemorrhagic diseases among the tick-borne viruses, and is the second most widespread of all medically critical arboviruses following dengue. Members of the Hyalomma genus are mainly involved in the natural transmission of the CCHF virus (CCHFV); of those, H. marginatum is known to be the primary vector of the disease in the Western Palaearctic. In general, epidemiological studies have been based on serological detections in the hosts and/or virus screening of ticks collected from the hosts. To the best of our knowledge, only a few studies have been carried out to screen the virus in unfed, questing field ticks. Nevertheless, detection of the virus in questing ticks is known to be a crucial parameter to determine the possible vector roles of the ticks and to understand the ecological dynamics of related diseases. In this study, 200 (75 males,125 females) questing H. marginatum adults collected from the field in nine villages in Thrace, located in the European part of Turkey, were screened individually for CCHFV using nested PCR. As a result, 103 (51.5 %) ticks were determined as positive with various strains of CCHFV. High positivity in questing vectors in a region where a significantly lower number of human cases have been encountered suggests that there should be some region-specific drivers that are effective in the natural dynamics of the disease. Detailed etiological and epidemiological studies are needed to reveal the possible reason for this unexpected discrepancy.

Crimean-Congo hemorrhagic fever virus in tortoises and Hyalomma aegyptium ticks in East Thrace, Turkey: potential of a cryptic transmission cycle.

BACKGROUND: Recent reports have demonstrated the presence of Crimean-Congo hemorrhagic fever virus (CCHFV) genomic material in Hyalomma aegyptium ticks feeding primarily on tortoises belonging to the genus Testudo. This raises the question if these ticks and their hosts play a role in the natural transmission dynamics of CCHFV. However, the studies are limited, and assessing the relevance of H. aegyptium in perpetuating the virus in nature, and a potential spillover to humans remains unknown. This study aimed to detect CCHFV in H. aegyptium ticks and their tortoise hosts in the East Thrace region of Turkey, where H. aegyptium is the most common human-biting tick and where a high density of tortoises of the genus Testudo can be found. METHODS: During the study period, 21 blood samples from different tortoises (2 T. hermanni and 19 T. graeca), 106 tick pools (containing 448 males, 152 females, 93 nymphs and 60 larvae) collected from 65 tortoises (5 T. hermanni and 60 T. graeca), 38 adult unfed questing ticks (25 males and 13 females, screened individually) and 14 pools (containing 8 nymphs and 266 larvae) of immature unfed questing ticks collected from the ground were screened for CCHFV genome by nested PCR and partial genomes sequenced. RESULTS: As a result of the screening of these 179 samples, 17 (9.5%) were detected as positive as follows: 2 of 21 blood samples (9.52%), 13 (containing 18 nymphs in 3 pools, and 52 males and 8 females in 10 pools) of 106 tick pools from tortoises (12.26%), and 2 of 38 adult questing ticks (5.26%). No positive result was determined in 14 pools of immature questing ticks. CONCLUSIONS: Previous studies have shown that reptiles can participate in the transmission of arthropod-borne viruses, but they may contribute to different aspects of the disease ecology and evolution of tick-borne viral pathogens. Our results indicate the presence of CCHFV in questing and feeding H. aegyptium ticks as well as tortoise hosts. This may indicate that CCHFV circulates in a cryptic transmission cycle in addition to the primary transmission cycle that could play a role in the natural dynamic of the virus and the transmission to humans.

Prevalence and molecular characterization of Giardia duodenalis in dogs in Aydin, Turkey.

The purpose of the present study was to investigate the prevalence and molecular characterization of Giardia duodenalis among dogs in Aydin, Turkey. A total of 473 faecal samples from dogs were collected. The overall prevalence of G. duodenalis was 18.8%. Higher infection rates were observed in dogs younger than three months old, from shelters, and with diarrhoea. Faecal samples of 89 dogs, diagnosed Giardia-positive by microscopy, were found positive by nested PCR. The ß-giardin nested PCR assay revealed assemblage B in all samples (100%), whereas 38 of the samples were mixed with assemblage A (42%). Sequence analysis of isolates indicated sub-genotypes A3 and B4 which have been previously detected in human isolates from Turkey. The results of the present study indicated the relatively high prevalence of giardiasis and the presence of the zoonotic sub-genotypes suggesting the important role of dogs as potential reservoirs of human infections.

External morphological anomalies in ixodid ticks from Thrace, Turkey.

Of 18,667 ticks examined, 33 specimens from species identified as Haemaphysalis parva, Hyalomma marginatum, Hy. scupense, Rhipicephalus bursa and Rh. turanicus were found to have external morphological anomalies. Anomalous Ha. parva, Hy. scupence and Rh. turanicus were reported in this study for the first time. General anomalies manifested as asymmetry and deformations of the idiosoma, whereas local anomalies occurred in legs, exoskeleton, spiracular, adanal, subanal and accessory plates, mouthparts and capitulum. With this study describing a gynandromorphic Hy. marginatum, the number of gynandromorphic tick cases has been raised to two in Turkey.

The morphological and behavioral analysis of geographically separated Rammeihippus turcicus (Orthoptera: Acrididae: Gomphocerinae) populations: data result in taxonomical conflict.

Rammeihippus Woznessenskij, 1996 (Orthoptera: Acrididae: Gomphocerinae) is a genus represented by two species. Rammeihippus turcicus (Ramme, 1939) is the only known species of the genus from Anatolia. As for most of the Gomphocerinae species in Anatolia, all populations of the species are intermittently distributed at high altitudes. In this study, three populations of R. turcicus were studied for the first time to determine the song and mating behavior. Males of the species produce typical calling song for Gomphocerinae and complex courtship songs and mating behavior. Thus, an accurate taxonomy requires extensive material and different character sources. In this study, the Anatolian Rammeihippus was re-examined on the basis of qualitative and morphometric morphology, male songs, and behavioral characteristics. There was no agreement between the results of the song and morphology. Acoustic analysis suggested one species and patchy distribution in the area, whereas morphology pointed out that each population was a different taxonomical unit. The results of the study show that the aberrant morphology does not necessarily indicate a new species in the Gomphocerinae genus.