Dermacentor Reticulatus Research Articles

Analysis of the species composition and infection of ticks with pathogens of Lyme disease in the territory of the Republic of Tatarstan for the period 2012–2022

BACKGROUND: The role of ixodid ticks in the transmission of pathogens of a number of natural focal infections can hardly be overestimated. Analysis of the manifestations of the epizootic process is important for planning preventive and anti-epidemic measures and is an integral component of epidemiological surveillance of natural focal infections. AIM: To analyze the species composition of ticks and the infection of ticks with borreliosis pathogens in the Republic of Tatarstan for the period 2012–2022. MATERIALS AND METHODS: An analysis of the species composition of ticks collected by the zoological group of the Center for Hygiene and Epidemiology in the Republic of Tatarstan from environmental objects during the period 2012–2022 was carried out. Testing of the ticks for Borrelia burgdorferi and Borrelia miyamotoi was carried out using the polymerase chain reaction method with hybridization-fluorescent detection. An analysis of statistical observation form No. 2 “Information on infectious and parasitic diseases” (the section “Tick bites”) for the Republic of Tatarstan from 2012 to 2022 was carried out. Statistical processing was performed using descriptive statistics methods in Microsoft Excel. RESULTS: The total number of people who applied for tick bites in the Republic of Tatarstan in 2012–2022 was 77,703, of which 59,168 (76.2%) were people over 18 years of age. During the study period, 5536 ticks were collected for research on the territory of the Republic of Tatarstan from environmental objects. The species composition of ticks was distributed as follows: Dermacentor reticulatus — 75.4%, Ixodes ricinus — 14.1%, Ixodes persulcatus — 10.5%. Infection of different types of ticks with Borrelia was not the same and was 20.4% for I. persulcatus, 14% for I. ricinus, 4.4% for D. reticulatus. In 4 regions of the republic, from ticks of the species I. ricinus pathogenic genomospecies B. miyamotoi were found (the infestation of this type of tick with B. miyamotoi was 0.4%). Indicators of morbidity of the population with ixodid tick-borne borreliosis and indicators of infection rates for ticks vary in different regions of the republic. CONCLUSIONS: The dominant species of ixodid ticks in natural epitopes of the Republic of Tatarstan in 2012–2022 were D. reticulatus (75.4% of the total number of collected ticks). For ticks of this species, the lowest level of infection with Borrelia was demonstrated (4.4% of ticks). This may be one of the factors responsible for the low, in comparison with other regions, incidence rates of tick-borreliosis in the Republic of Tatarstan. The identification of ticks of the species I. ricinus infected with B. miyamotoi confirms the need for targeted molecular genetic and serological examination for recurrent fever in patients with non-erythematous forms of febrile illness after an episode of tick bite.

Machine learning algorithms for the evaluation of risk by tick-borne pathogens in Europe

Background Tick-borne pathogens pose a major threat to human health worldwide. Understanding the epidemiology of tick-borne diseases to reduce their impact on human health requires models covering large geographic areas and considering both the abiotic traits that affect tick presence, as well as the vertebrates used as hosts, vegetation, and land use. Herein, we integrated the public information available for Europe regarding the variables that may affect habitat suitability for ticks and hosts and tested five machine learning algorithms (MLA) for predicting the distribution of four prominent tick species across Europe. Materials and methods A grid of cells 20 km in diameter was prepared to cover the entire territory, containing data on vegetation, points of water, habitat fragmentation, forest density, grass extension, or imperviousness, with information on temperature and water deficit. The distribution of the hosts (162 species) was modelled and included in the dataset. We used five MLA, namely, Random Forest, Neural Networks, Naive Bayes, Gradient Boosting, and AdaBoost, trained with reliable coordinates for Ixodes ricinus, Dermacentor reticulatus, Dermacentor marginatus, and Hyalomma marginatum in Europe. Results Both Random Forest and Gradient Boosting best predicted ticks and host environmental niches. Our results demonstrate that MLA can identify trait-matching combinations of environmental niches. The inclusion of land cover and land use variables has a superior capacity for predicting areas suitable for ticks, compared to classic methods based on the use of climate data alone. Conclusions Flexible MLA-driven models may offer several advantages over traditional models. We anticipate that these results may be extrapolated to other regions and combinations of tick-vertebrates. These results highlight the potential of MLA for inference in ecology and provide a background for the evolution of a completely automatized tool to calculate the seasonality of ticks for early warning systems aimed at preventing tick-borne diseases.

The prevalence of pathogens in ticks collected from humans in Belgium, 2021, versus 2017

BackgroundTicks carry a variety of microorganisms, some of which are pathogenic to humans. The human risk of tick-borne diseases depends on, among others, the prevalence of pathogens in ticks biting humans. To follow-up on this prevalence over time, a Belgian study from 2017 was repeated in 2021.MethodsDuring the tick season 2021, citizens were invited to have ticks removed from their skin, send them and fill in a short questionnaire on an existing citizen science platform for the notification of tick bites (TekenNet). Ticks were morphologically identified to species and life stage level and screened using multiplex qPCR targeting, among others, Borrelia burgdorferi (sensu lato), Anaplasma phagocytophilum, Borrelia miyamotoi, Neoehrlichia mikurensis, Babesia spp., Rickettsia helvetica and tick-borne encephalitis virus (TBEV). The same methodology as in 2017 was used.ResultsIn 2021, the same tick species as in 2017 were identified in similar proportions; of 1094 ticks, 98.7% were Ixodes ricinus, 0.8% Ixodes hexagonus and 0.5% Dermacentor reticulatus. A total of 928 nymphs and adults could be screened for the presence of pathogens. Borrelia burgdorferi (s.l.) was detected in 9.9% (95% CI 8.2–12.0%), which is significantly lower than the prevalence of 13.9% (95% CI 12.2–15.7%) in 2017 (P = 0.004). The prevalences of A. phagocytophilum (4.7%; 95% CI 3.5–6.3%) and R. helvetica (13.3%; 95% CI 11.2–15.6%) in 2021 were significantly higher compared to 2017 (1.8%; 95% CI 1.3–2.7% and 6.8%; 95% CI 5.6–8.2% respectively) (P < 0.001 for both). For the other pathogens tested, no statistical differences compared to 2017 were found, with prevalences ranging between 1.5 and 2.9% in 2021. Rickettsia raoultii was again found in D. reticulatus ticks (n = 3/5 in 2021). Similar to 2017, no TBEV was detected in the ticks. Co-infections were found in 5.1% of ticks. When combining co-infection occurrence in 2017 and 2021, a positive correlation was observed between B. burgdorferi (s.l.) and N. mikurensis and B. burgdorferi (s.l.) and B. miyamotoi (P < 0.001 for both).ConclusionsAlthough the 2021 prevalences fell within expectations, differences were found compared to 2017. Further research to understand the explanations behind these differences is needed.Graphical

Збудники інфекційних хвороб людини і тварин та їх розповсюдження в Європі іксодовим кліщем Dermacentor reticulatus

Due to climate change, the possibility of human infective diseases caused by pathogens from the genera Borelia, Anaplasma, Rickettsia, Babesia, Bartonella, Coxiella, etc. is increasing in Europe. Generally, these pathogens are directly related to arthropod vectors, in particular, to Ixodes ticks. The aim of the study was to determine the current trends in the spread of tick-borne infections transmitted by Dermacentor reticulatus ticks in some European countries, including Ukraine, based on the analysis of literature. Over the past 10 years, in the vast majority of countries, including Europe, the situation with the defeat of farm and wild animals, as well as humans, by pathogens of severe infectious diseases from the genera Borrelia, Anaplasma, Rickettsia, Babesia, Theileria, Coxiella, Bartonella, Francisella, etc. has been aggravated. Representatives of these genera lead to severe diseases, such as tick-borne borreliosis, which are accompanied by damage to the central nervous system, joints, skin, and cardiovascular system. Members of the Rickettsiales genus cause a variety of feverish conditions in animals and humans. Species of the genus Babesia cause babesiosis, a disease caused by damage to red blood cells and the appearance of anti-erythrocyte antibodies, and representatives of the genus Toxoplasma cause toxoplasmosis, accompanied by damage to the nervous, lymphatic, and cardiovascular systems. Q fever is caused by species of the genus Coxiella. Severe human diseases are caused by tick-borne encephalitis viruses. Various PCR test systems have been developed and used to detect these pathogens. For example, in Poland, PCR for Borrelia burgdorferi s.l. was performed using the B. burgdorferi s.l. PCR kit (GeneProof, Czech Republic) for in vitro diagnosis. Anaplasma PCR kits (Blirt-DNA Gdańsk, Poland) were used to detect Anaplasma phagocytophilum DNA. For the detection of Babesia spp., a fragment of the 18S rDNA gene encoding a small ribosomal subunit localized in the conserved region V4 was used. To detect Bartonella spp., the tests Hum PCR Bartonella, Hum PCR Coxiella burnetii, and Vet PCR Rickettsia kit (BioIngenTech, Chile), Coxiella burnetii and Rickettsia spp. were used. In the Laboratory for the Study of Ticks and Tick-Borne Infections of Ternopil National Medical University (Ukraine), pathogens were identified using several test systems, namely DNA A. phagocytophilum, / DNA E. muris, / DNA E. chaffeensis, DNA B. burgdorferi s.l., DNA B. myamotoi; DNA Babesia sp., and RNA Ticks borne of encephalitis viruses (MolGen, Italy), which allow real-time determination on anamplifier "ROTOR Gene 6000" by Corbett Research. Studies in many European countries have shown infection of the Dermacentor reticulatus tick with the pathogens B. burgdorferi s.l. in the range from 1.6 to 49.8 %; A. phagocytophilum – 0.1–2.2 %; Babesia sрp. – 9.8–43.8 %; Coxiella spр. – 0.1–0.2%; Francisella spp. – 0.7–95%; Toxoplasma spр. – 3.2–16.7 %; and ticks borne of encephalitis viruses 8.5–16.6 %. Some Ukrainian scientists presented data on D. reticulatus infection, in particular B. burgdorferi s. l. – 10.1–22.1 %; A. phagocytophilum – 3–25.36 %; Babesia spр. – 1–41.2 %; Rickettsia sp. – 28 %; Bartonella spp. – 5 %; Coxiella spр. – 5.1 %, and tick-borne f encephalitis viruses – 11.9–71 %. The spread of many infectious agents is largely correlated with the spread of their vectors, including ticks of the D. reticulatus species. According to many scientists, this species is widespread in Europe, in particular, two metapopulations are currently distinguished as the western one, which includes the territories of France, Belgium, Slovakia, the Czech Republic, the Netherlands, and Germany, and the eastern one, which covers Lithuania, Latvia, Belarus, eastern and central Poland, and west of the Vistula River up to the Ural Mountains. In the Czech Republic, this species is found in all regions, more often in the southeastern and northwestern regions. In the UK, ticks have been found in West Wales, Devon, and Essex. In Romania, D. reticulatus was found in 17 counties, and in Moldova – in the Lower Dniester region. In Ukraine, this species is widespread from the Carpathians and Prykarpattia, Polissya, and Forest-Steppe, to the southeastern regions and Crimea.

A New Endemic Locality of Dermacentor reticulatus in Central-Southern Poland and Its Potential Epidemiological Implications.

Dermacentor reticulatus (Acari: Ixodidae) is an important arthropod vector in medical and veterinary contexts. Its geographic range is divided into western and eastern populations separated by a "Dermacentor-free zone" in central Poland. Recent faunistic studies showed a new endemic locality of the species in Upper Silesia to the west of the Vistula River (central-southern Poland) and its co-occurrence with I. ricinus. The prevalence of five tick-borne pathogens (TBPs), e.g., B. burgdorferi s.l., Bartonella spp., Rickettsia spp., and Babesia spp., in the ticks was assessed with polymerase chain reaction (PCR) methods. The molecular studies revealed the presence of Rickettsia spp. in 23.8% of the D. reticulatus specimens. In turn, 94.1% of the I. ricinus adults were infected with B. burgdorferi s.l., 11.7 % with Babesia spp., and 5.8% with Rickettsia spp. Coinfections with two TBPs were noted in 17.6% of the I. ricinus. These findings highlight not only the risk of infestation by both tick species in an area previously considered Dermacentor-free, but also the high prevalence of TBPs in the study area. Increased focus on medical and veterinary services appears necessary to diagnose and prevent tick-borne diseases in this region.

Tick-Borne pathogens and defensin genes expression: A closer look at Ixodes ricinus and Dermacentor reticulatus

The immune system of ticks, along with that of other invertebrates, is comparatively simpler than that of vertebrates, relying solely on innate immune responses. Direct antimicrobial defence is provided by the synthesis of antimicrobial peptides (AMPs), including defensins. The aim of this study was to investigate the differences in defensin genes expression between questing and engorged Ixodes ricinus (def1 and def2) and Dermacentor reticulatus (defDr) ticks, in the presence of selected pathogens: Borrelia spp., Rickettsia spp., Babesia spp., Anaplasma phagocytophilum, and Neoehrlichia mikurensis in the natural environment. After pathogen screening by PCR/qPCR, the expression of defensin genes in pathogen positive ticks and ticks without any of the tested pathogens, was analysed by reverse transcription qPCR. The results showed an increased expression of defensin genes in I. ricinus ticks after blood feeding and I. ricinus and D. reticulatus ticks during in cases of co-infection. In particular, the expression of defensins genes was higher in questing D. reticulatus than in questing and engorged I. ricinus ticks, when borreliae were detected. This study contributes to uncovering the expression patterns of defensin genes in the presence of several known tick pathogens, the occurrence of these pathogens and possible regulatory mechanisms of defensins in tick vector competence.

Ticks and spirochetes of the genus Borrelia in urban areas of Central-Western Poland.

Due to the extensive use of green urban areas as recreation places, city residents are exposed to tick-borne pathogens. The objectives of our study were (i) to determine the occurrence of ticks in urban green areas, focussing on areas used by humans such as parks, schools and kindergartens, and urban forests, and (ii) to assess the prevalence of Borrelia infections in ticks in Zielona Góra, a medium-sized city in western Poland. A total of 161 ticks representing the two species Ixodes ricinus (34 males, 51 females, 30 nymphs) and Dermacentor reticulatus (20 males, 26 females) were collected from 29 of 72 (40.3%) study sites. In total, 26.1% of the ticks (85.7% of I. ricinus and 14.3% of D. reticulatus) yielded DNA of Borrelia. The difference in the infection rate between I. ricinus and D. reticulatus was significant. Among infected ticks, the most frequent spirochete species were B. lusitaniae (50.0%) and B. afzelii (26.2%), followed by B. spielmanii (9.5%), B. valaisiana (7.1%), B. burgdorferi sensu stricto, (4.8%) and B. miyamotoi (2.4%). No co-infections were found. We did not observe a correlation in the occurrence of Borrelia spirochetes in ticks found in individual study sites that differed in terms of habitat type and height of vegetation. Our findings demonstrate that the Borrelia transmission cycles are active within urban habitats, pointing the need for monitoring of tick-borne pathogens in public green areas. They could serve as guidelines for authorities for the proper management of urban green spaces in a way that may limit tick populations and the potential health risks posed by tick-borne pathogens.

Tick hazard in the South Downs National Park (UK): species, distribution, key locations for future interventions, site density, habitats.

South Downs National Park (SDNP) is UK's most visited National Park, and a focus of tick-borne Lyme disease. The first presumed UK autochthonous cases of tick-borne encephalitis and babesiosis were recorded in 2019-20. SDNP aims to conserve wildlife and encourage recreation, so interventions are needed that reduce hazard without negatively affecting ecosystem health. To be successful these require knowledge of site hazards. British Deer Society members submitted ticks removed from deer. Key potential intervention sites were selected and six 50 m2 transects drag-sampled per site (mostly twice yearly for 2 years). Ticks were identified in-lab (sex, life stage, species), hazard measured as tick presence, density of ticks (all life stages, DOT), and density of nymphs (DON). Sites and habitat types were analysed for association with hazard. Distribution was mapped by combining our results with records from five other sources. A total of 87 Ixodes ricinus (all but one adults, 82% F) were removed from 14 deer (10 Dama dama; three Capreolus capreolus; one not recorded; tick burden, 1-35) at 12 locations (commonly woodland). Five key potential intervention sites were identified and drag-sampled 2015-16, collecting 623 ticks (238 on-transects): 53.8% nymphs, 42.5% larvae, 3.7% adults (13 M, 10 F). Ticks were present on-transects at all sites: I. ricinus at three (The Mens (TM); Queen Elizabeth Country Park (QECP); Cowdray Estate (CE)), Haemaphysalis punctata at two (Seven Sisters Country Park (SSCP); Ditchling Beacon Nature Reserve (DBNR)). TM had the highest DOT at 30/300 m2 (DON = 30/300 m2), followed by QECP 22/300 m2 (12/300 m2), CE 8/300 m2 (6/300 m2), and SSCP 1/300 m2 (1/300 m2). For I. ricinus, nymphs predominated in spring, larvae in the second half of summer and early autumn. The overall ranking of site hazard held for DON and DOT from both seasonal sampling periods. DBNR was sampled 2016 only (one adult H. punctata collected). Woodland had significantly greater hazard than downland, but ticks were present at all downland sites. I. ricinus has been identified in 33/37 of SDNPs 10 km2 grid squares, Ixodes hexagonus 10/37, H. punctata 7/37, Dermacentor reticulatus 1/37. Mapping shows tick hazard broadly distributed across SDNP. I. ricinus was most common, but H. punctata's seeming range expansion is concerning. Recommendations: management of small heavily visited high hazard plots (QECP); post-visit precaution signage (all sites); repellent impregnated clothing for deerstalkers; flock trials to control H. punctata (SSCP, DBNR). Further research at TM may contribute to knowledge on ecological dynamics underlying infection density and predator re-introduction/protection as public health interventions. Ecological research on H. punctata would aid control. SDNP Authority is ideally placed to link and champion policies to reduce hazard, whilst avoiding or reducing conflict between public health and ecosystem health.

Study of ixodid ticks in recreational areas of large cities in 2017–2022

Background. Ixodes ticks are generalist ticks, which can easily adapt to new conditions, and therefore are widespread in urban areas. They are central components for understanding tick-borne disease epidemiology in large cities. Materials and methods. The main methods of collecting ticks were flagging, which was most often used; ticks were also collected manually by removing them from human and animal bodies. Ticks were examined in the laboratory by dark-field microscopy using optoelectronic systems IMAGLAV-SEO, Leica DM 500 (204), Olimpus, ZEISS Axio Lab A1, and MIKMED-1. PCR studies were performed in real time on a ROTOR Gene 6000 amplifier. Results. According to the results of field and laboratory studies in the parks of Ternopil during 2018–2019 by the Ternopil Regional Center for Disease Control and Prevention (TRCDCP) of the Ministry of Health of Ukraine, 524 ticks were collected, 515 of which were examined using dark-field microscopy (DFM) and polymerase chain reaction (PCR). Among the collected specimens, 63 (12.23 %) were the carriers of infectious pathogens, including 24 (4.66 %) carriers of Borrelia burgdorferi s.l. (Johnson et al., 1984). Besides, 7654 ticks removed from humans were examined in the laboratory of the TRCDCP and in the laboratory of the I. Horbachevsky Ternopil National Medical University. Of these, 1280 (16.72 %) specimens were carriers of infectious agents. The Lviv CDC collected 1314 ticks in recreational areas of Lviv during 2017-2022, of which 139 (10.58 %) were vectors. Of the 1136 ticks collected from humans, 295 (25.96 %) were carriers of B. burgdorferi s.l. Out of 540 ticks collected in parks by the Chernivtsi Regional Center for Tick Control in Chernivtsi, 164 (30.37 %) were infected with B. burgdorferi s.l. Out of 454 ticks collected from people in the city, 130 (28.63 %) were infected with B. burgdorferi s.l. In the parks of Khmelnytskyi, researchers of the Khmelnytskyi Regional Center for Tick Control collected 5068 tick species, of which 102 (2.01 %) were carriers of B. burgdorferi s.l. Spirochetes of B. burgdorferi s.l. were detected in 694 (34.87 %) species out of 1990 examined ticks removed from humans. Conclusions. The dominant tick species in the recreational areas of Lviv, Ternopil, Chernivtsi and Khmelnytsky regions are Ixodes ricinus and Dermacentor reticulatus. The proportion of ticks collected in city parks that carry infectious agents ranges from 10.58 % to 30.37 %, with the exception of Khmelnytsky, where the tick infestation rate is only 2.01 %. The infection rate of ticks removed from humans with B. burgdorferi s.l. and other pathogens ranges from 16.72 % to 34.87 %.

The efficacy of antiparasitic drug based on Lufenuron, Moxidectin, and Praziquantel against ixodidosis in dogs

The purpose of the research is to study the efficacy of antiparasitic drug based on Lufenuron, Moxidectin, Praziquantel against ixodidosis in dogs under natural conditions and during experimental infection with ixodid ticks.Materials and methods. The experiments used the drug in tablet formulation containing Lufenuron in 320 mg, Moxidectin in 9.6 mg and Praziquantel in 160 mg. To study the preventive and therapeutic acaricidal effect of the drug against ixodid ticks in the country part of the Karachaevsky District of the Karachay-Cherkess Republic, two groups (10 dogs each) were formed of Alabai dogs or outbred dogs with a body weight of 16–76 kg. The dogs of the first experimental group were administered the drug in tablets. The dogs of the second group served as controls and were not treated. The experiment used clinically healthy dogs of both sexes over 6 weeks of age, spontaneously infected with Ixodidae. Animals were selected so that they were not exposed to drugs for at least 1 month prior to the experiment. The animals were kept under normal conditions throughout the experiment and received their usual food. Antiparasitic tablets were administered to the dogs individually, once, orally, mixed with food at a dose of 1 tablet per 16–32 kg of body weight. The ixodid infection was diagnosed in the dogs by clinical examination. The genus of Ixodidae was determined by typical morphological characteristics. The drug efficacy was evaluated based on reduced number or absence of ticks on the treated animals (experimental) versus the untreated animals (control). Before the experiment and after the drug, the animals were examined every hour for 48 hours, and condition, separation, separation time from the start of the drug and death of the ticks were recorded; the dogs were then examined every 5–7 days for two months.Results and discussion. The article presents study data on antiparasitic drug efficacy against ixodidosis in dogs spontaneously infected with Ixodes ricinus, I. persulcatus, and Dermacentor reticulatus. The result of the studies found that the antiparasitic drug (active substances were Lufenuron, Moxidectin, and Praziquantel) had a therapeutic effect that was recorded on day 5 after treatment and was 90%. The preventive efficacy was also above 70%: on day 7 and 14, this value was 90.0 and 86.9%, respectively, which indicated the duration of the preventive effect for 25–28 days. To fully evaluate the acaricidal efficacy against ixodidosis, studies were conducted on experimentally infected animals. Forty percent efficacy was determined at 48 hours after the drug; at 2 weeks after the drug, 90 %, 98.5 and 100 % efficacy were determined against ixodidosis in the dogs at 24 hours, 48 and 72 hours after the transfer of ticks, respectively. At 4 weeks after the drug, 100 % efficacy was determined against ixodidosis in the dogs at 24, 48 and 72 hours after the transfer of ticks, respectively.

Tick-Borne Pathogens in Dermacentor reticulatus Ticks from Bosnia and Herzegovina.

Dermacentor (D.) reticulatus ticks carry and transmit a wide range of pathogens to vertebrate hosts. Limited information is available about the existence of emerging tick-borne pathogens and the distribution of D. reticulatus in Bosnia and Herzegovina. The study aimed to investigate the occurrence and distribution of D. reticulatus and to detect the presence of Anaplasma spp., Borrelia (B.) burgdorferi s.l., Rickettsia spp., and Babesia spp. in samples originating from questing ticks and ticks collected from domestic animals in various regions of Bosnia and Herzegovina. A total of 402 collected D. reticulatus ticks were widely distributed throughout the country. Of the 41 pools consisting of 205 individual D. reticulatus ticks, 21 (51.2%) indicated the presence of Rickettsia spp., 17 (41.4%) of Babesia spp., 2 (4.8%) of Anaplasma spp., and 1 (2.4%) of B. burgdorferi s.l. after real-time PCR screening. Our study indicates that D. reticulatus has significantly expanded its distribution and host range in Bosnia and Herzegovina. Moreover, our results represent the first detection of Babesia spp. in D. reticulatus in Bosnia and Herzegovina. Given the demonstrated presence of emerging pathogens in questing and feeding ticks, there is an urge to establish a surveillance system for ticks and tick-borne pathogens in Bosnia and Herzegovina.

Genotyping of Borrelia, Rickettsia and Anaplasma in Ixodes ricinus and Dermacentor reticulatus ticks in the Kaliningrad region

Background. Tick-borne bacterial and protozoal pathogens pose a significant public health problem. The aim of this study was to detect and genotype Borrelia, Rickettsia and Anaplasma in Ixodes ricinus and Dermacentor reticulatus ticks collected in the Kaliningrad region in 2021–2022. Materials and methods. The study included 862 I. ricinus and 803 D. reticulatus ticks (1665 in total) collected in 33 biotopes of the Kaliningrad region. Detection of the DNA of tick-borne pathogens was carried out in individual ticks by PCR using a set of specific primers, followed by sequencing and phylogenetic analysis. Results. The level of infection of I. ricinus ticks with Borrelia was 15.5%, and genotyping by the p66 gene sequence showed the presence of genetic material from four species: B. afzelii, B. garinii, B. valaisiana, and B. lusitaniae. In D. reticulatus ticks, no Borrelia genetic material was detected. The Rickettsia DNA has been found in both tick species. Moreover, the infection rate of I. ricinus ticks was 2.6%, and D. reticulatus — 21.2%. R. helvetica were found in I. ricinus ticks, and R. raoultii in meadow ticks when genotyping by gltA gene. Genetic markers of Anaplasma phagocytophilum have been found in I. ricinus and D. reticulatus ticks. Cases of co-infection of an individual tick have also been identified. Conclusion. Six different species of tick-borne pathogens were found in the I. ricinus and D. reticulatus ticks collected in the Kaliningrad region and R. helvetica, R. raoultii and A. phagocytophilum were identified for the first time.

Spatial Distribution and Pathogen Profile of Dermacentor reticulatus Ticks in Southeastern Poland: A Genetic and Environmental Analysis

In recent years, significant changes have been observed in the distribution and abundance of local Dermacentor reticulatus populations. However, changes in D. reticulatus dynamics have not been studied in southeastern Poland. Our objective was to enhance our understanding of the environmental factors influencing the occurrence and density of D. reticulatus in this area. Additionally, we sought to investigate the genetic diversity of the tick population and the prevalence of tick-borne pathogens (TBPs). To this end, we established 45 study sites in the Subcarpathian province. Ticks were collected during their peak activity in both spring and autumn. A subset of randomly selected specimens underwent molecular analysis for TBPs screening, using high-throughput microfluidic real-time PCR. Positive amplicons were then sequenced, and phylogenetic analyses were conducted. Our findings confirmed the presence of D. reticulatus ticks in 24 surveyed sites, primarily concentrated in the northern and eastern parts of the region. The mean density of D. reticulatus ticks in their compact range was 5.8 ± 6.4 specimens/100 m2. Notably, air temperature and altitude emerged as significant factors influencing the species’ activity. We also identified a high prevalence of Rickettsia raoultii infections in adult D. reticulatus, reaching up to 84.21%. Additionally, 9.52% of ticks were found to be infected with R. helvetica and 4.76% with Anaplasma phagocytophilum. Furthermore, our genetic analyses confirmed the identity of D. reticulatus in the Subcarpathian region, aligning with haplotypes found in other regions of Poland, Czechia, Croatia, and Portugal. In conclusion, our study suggests that the surveyed region represents the current boundary of the compact range of D. reticulatus in Poland in which this tick species exhibits low genetic diversity and a narrow spectrum of detected TBPs.

Identification of New Microfoci and Genetic Characterization of Tick-Borne Encephalitis Virus Isolates from Eastern Germany and Western Poland.

(1) Background: Tick-borne encephalitis (TBE) is the most important tick-borne viral disease in Eurasia, although effective vaccines are available. Caused by the tick-borne encephalitis virus (TBEV, syn. Orthoflavivirus encephalitidis), in Europe, it is transmitted by ticks like Ixodes ricinus and Dermacentor reticulatus. TBEV circulates in natural foci, making it endemic to specific regions, such as southern Germany and northeastern Poland. Our study aimed to identify new TBEV natural foci and genetically characterize strains in ticks in previously nonendemic areas in Eastern Germany and Western Poland. (2) Methods: Ticks were collected from vegetation in areas reported by TBE patients. After identification, ticks were tested for TBEV in pools of a maximum of 10 specimens using real-time RT-PCR. From the positive TBEV samples, E genes were sequenced. (3) Results: Among 8400 ticks from 19 sites, I. ricinus (n = 4784; 56.9%) was predominant, followed by D. reticulatus (n = 3506; 41.7%), Haemaphysalis concinna (n = 108; 1.3%), and I. frontalis (n = 2; <0.1%). TBEV was detected in 19 pools originating in six sites. The phylogenetic analyses revealed that TBEV strains from Germany and Poland clustered with other German strains, as well as those from Finland and Estonia. (4) Conclusions: Although there are still only a few cases are reported from these areas, people spending much time outdoors should consider TBE vaccination.

Novel clades of tick-borne pathogenic nairoviruses in Europe

Members of the Orthonairovirus genus (family Nairoviridae) include many tick-borne viruses of significant human and animal health impact, with several recently-documented pathogenic viruses lacking sufficient epidemiological information. We screened 215 adult ticks of seven species collected in Bulgaria, Georgia, Latvia and Poland for orthonairoviruses, followed by nanopore sequencing (NS) for genome characterization. Initial generic amplification revealed Sulina virus (SULV, Orthonairovirus sulinaense), for which an updated amplification assay was used, revealing an overall prevalence of 2.7% in Ixodes ricinus ticks from Latvia. Three complete and additional partial SULV genomes were generated, that consistently formed a separate, distinct clade with further intragroup divergence in the maximum likelihood analyses. Comparisons with previously described viruses from Romania exhibited similar genome topologies, albeit with divergent motifs and cleavage sites on the glycoprotein precursor. Preliminary evidence of recombination involving the S segment was documented, in addition to variations in predicted viral glycoproteins. Generic screening further identified Tacheng tick virus 1 (TCTV1, Orthonairovirus tachengense), with documented human infections, in Dermacentor reticulatus ticks from Poland, with a prevalence of 0.9%. Subsequent NS and assembly provided the first complete TCTV1 genome outside of China, where it was originally described. Phylogenetic analysis of virus genome segments revealed TCTV1-Poland as a discrete taxon within the TCTV1 cluster in the Orthonairovirus genus, representing a geographically segregated clade. Comparable genome topology with TCTV1 from China was observed, aside from minor variations in the M segment. Similar to SULV, TCTV1 exhibited several mismatches on previously described screening primer binding sites, likely to prevent amplification. These findings indicate presence of novel TCTV1 and SULV clades in Eastern Europe, confirming the expansion of orthonairoviruses with pathogenic potential.

Investigations of the tick burden on passeriform, water-associated and predatory birds reveal new tick-host associations and habitat-related factors of tick infestation.

Previous studies on the tick infestation of birds in the Carpathian Basin focused on songbirds (Passeriformes). Thus, the primary aim of the present work was to extend the scope of previous studies, i.e. to include aquatic (water-associated) bird species in a similar context, especially considering that these birds are usually long-distance migrants. Between March 2021 and August 2023, 11,919 birds representing 126 species were checked for the presence of ticks. From 352 birds belonging to 40 species, 905 ixodid ticks were collected. Tick species were identified morphologically and/or molecularly. Ticks from avian hosts belonged to seven species: Ixodes ricinus (n = 448), I. frontalis (n = 31), I. festai (n = 2), I. arboricola (n = 36), I. lividus (n = 4), Haemaphysalis concinna (n = 382) and Dermacentor reticulatus (n = 2). Nymphs of I. ricinus occurred with a single activity peak around March-May, whereas its larvae typically infested birds in May, June or July. By contrast, H. concinna usually had its activity maximum during the summer (nymphs in June-July, larvae later in July-August). Interestingly, two ornithophilic species, I. frontalis and I. arboricola, were most active around winter months (between October and April). A significantly lower ratio of aquatic birds was found tick-infested than songbirds. Several new tick-host associations were revealed, including I. ricinus from Greylag Goose (Anser anser) and D. reticulatus from Great Egret (Ardea alba) and Sedge Warbler (Acrocephalus schoenobaenus). Ticks were collected for the first time in Europe from two species of predatory birds as well as from Little Bittern (Ixobrychus minutus). Bird species typically inhabiting reedbeds were most frequently infested with H. concinna, and most ticks localized at their throat, as opposed to forest-dwelling avian hosts, on which I. ricinus predominated and ticks were more evenly distributed. In the evaluated region, aquatic birds appear to be less important in tick dispersal than songbirds. However, newly revealed tick-host associations in this category attest to their hitherto neglected contribution. The results suggest that the habitat type will have significant impact not only on the species composition but also on the feeding location of ticks on birds.

Application of DNA barcodes in the genetic diversity of hard ticks (Acari: Ixodidae) in Kazakhstan.

Forty-five tick species have been recorded in Kazakhstan. However, their genetic diversity and evolutionary relationships, particularly when compared to ticks in neighbouring countries, remain unclear. In the present study, 148 mitochondrial cytochrome c oxidase subunit I (COI) sequence data from our laboratory and NCBI (National Center for Biotechnology Information; https://www.ncbi.nlm.nih.gov/ ) data were used to address this knowledge gap. Phylogenetic analyses showed that i) Hyalomma anatolicum anatolicum (Koch, 1844) ticks from Jambyl Oblast (southeastern Kazakhstan) and Gansu Province (northwestern China) constituted a newly deviated clade; and ii) Dermacentor reticulatus (Fabricius, 1974) ticks from South Kazakhstan Oblast were closer to those in Romania and Turkey. The network diagram of haplotypes showed that i) the H-1 and H-2 haplotypes of Dermacentor marginatus (Sulzer, 1776) ticks from Zhetisu and Almaty were all newly evolved; and ii) the H-3 haplotypes of Haemaphysalis erinacei (Pavesi, 1884) from Almaty Oblast and Xinjiang Uygur Autonomous Region (northwestern China) were evolved from the H-1 haplotype from Italy. In the future, more COI data from different tick species, especially from Kazakhstan and neighbouring countries, should be employed in the field of tick DNA barcoding.

Mixed-sex clusters on grass blades: breeding strategy of the ornate dog tick, Dermacentor reticulatus

BackgroundThe ornate dog tick Dermacentor reticulatus is second only to the hard tick Ixodes ricinus in terms of importance as a vector of infectious organisms, especially of Babesia canis, the agent of canine babesiosis. Both the geographical range and local densities of D. reticulatus are steadily increasing in many regions of Europe. In the present study, we tested the hypothesis that D. reticulatus possesses an efficient breeding strategy that allows for a rapid increase in tick numbers and densities through the formation of mixed-sex clusters/aggregations while questing in the environment.MethodsAn observational study was carried out in the spring of 2023, at three sites in two regions in Central and North-Eastern Poland, both characterised by high tick densities. At each site, a 400-m-long transect was inspected for questing ticks. All noted ticks were collected, and tick numbers and sexes per stem were recorded. Differences in tick distribution by site and sex were analysed statistically.ResultsA total of 371 D. reticulatus (219 females, 152 males) ticks were collected from 270 grass stems over a combined 1200 m of transect. The majority of grass stems (74.4%) were occupied by just a single individual, with two-tick clusters the second most common category. The maximum number of D. reticulatus individuals observed on a single grass stem was six.Mixed-sex clusters were significantly more common than single-sex clusters at all three sites. With study sites combined, mixed-sex clusters accounted for 17.4% (95% confidence limit [95% CL] 13.9–21.6%) of observations, while for multiple males and multiple females, the values were 2.6% (95% CL: 1.4–4.7%) and 5.6% (95% CL: 3.7–8.3%), respectively.ConclusionsMixed-sex clusters of D. reticulatus ticks were significantly more common than single-sex clusters, which we hypothesise reflects an efficient, likely pheromone-mediated breeding strategy of this expansive tick species.Graphical

Phylogenetic characterisation of tick-borne encephalitis virus from Lithuania.

The Baltic states are the region in Europe where tick-borne encephalitis (TBE) is most endemic. The highest notification rate of TBE cases is reported in Lithuania, where the incidence of TBE has significantly increased since 1992. A recent study reported 0.4% prevalence of TBE virus (TBEV) in the two most common tick species distributed in Lithuania, Ixodes ricinus and Dermacentor reticulatus, with the existence of endemic foci confirmed in seven out of Lithuania's ten counties. However, until now, no comprehensive data on molecular characterisation and phylogenetic analysis have been available for the circulating TBEV strains. The aim of this study was to analyse TBEV strains derived from I. ricinus and D. reticulatus ticks collected from Lithuania and provide a genotypic characterisation of viruses based on sequence analysis of partial E protein and NS3 genes. The 54 nucleotide sequences obtained were compared with 81 TBEV strains selected from the NCBI database. Phylogenetic analysis of the partial E and NS3 gene sequences derived from 34 Lithuanian TBEV isolates revealed that these were specific to Lithuania, and all belonged to the European subtype, with a maximum identity to the Neudoerfl reference strain (GenBank accession no. U27495) of 98.7% and 97.4%, respectively. The TBEV strains showed significant regional genetic diversity. The detected TBEV genotypes were not specific to the tick species. However, genetic differences were observed between strains from different locations, while strains from the same location showed a high similarity.

Evaluation of the efficacy of Simparica Trio® in the prevention of the transmission of Babesia canis by infected Dermacentor reticulatus to dogs

BackgroundBabesia canis is a clinically relevant vector-borne pathogen in dogs, and its presence is expanding. The efficacy of Simparica Trio® (Zoetis) in the prevention of B. canis transmission was evaluated at the minimum recommended label dose of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin and 5 mg/kg pyrantel per kg bodyweight.MethodsTwenty-four (24) dogs were randomly allocated to either a placebo-treated group or one of two treatment groups with Simparica Trio. Dogs were infested with B. canis-infected Dermacentor reticulatus ticks 21 or 28 days after treatment administration. Blood samples for antibody and DNA detection were collected from each dog prior to tick infestation until 28 days after infestation. A dog was defined as being B. canis positive if it tested positive by both an indirect immunofluorescence assay (IFA) and PCR at any time during the study.ResultsNo treatment-related adverse reactions were recorded during the study. All placebo-treated animals displayed clinical signs due to babesiosis and tested positive on both IFA and PCR. None of the Simparica Trio-treated animals displayed any clinical symptoms or tested positive, resulting in a 100% efficacy in the prevention of canine babesiosis (P < 0.0001).ConclusionsA single treatment with Simparica Trio at the minimum recommended label dose of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin and 5 mg/kg pyrantel per kg bodyweight prevents the transmission of B. canis by infected D. reticulatus to dogs for at least 28 days.Graphical