A First Look at the Relationship Between Large Herbivore-Induced Landscape Modifications and Ixodes ricinus Tick Abundance in Rewilding Sites.

Background: While the influence of landscape and microclimatic conditions on tick populations is well-documented, there remains a gap in more specific data regarding their relationship to rewilding efforts with large herbivore activity. Objective: This pilot study, spanning from 2019 to 2021, explores the effects of naturalistic grazing by large semi-wild ungulates on tick abundance in the Milovice Reserve, Czechia. Methods: Tick collection was observed using flagging techniques at two distinct sites of rewilding area: one grazed, actively utilized by animals involved in the rewilding project, and one ungrazed, left fallow in neighboring areas utilized only by wild animals. Transects, each measuring 150 m in length and 5 m in width (750 m2), were established at these two sampling locations from March to September between 2019 and 2021. To minimize potential bias resulting from tick movement, a 300 m buffer zone separated the two sites. Data analysis employed a generalized estimating equations (GEE) model with negative binomial regression. The study assessed potential variations in tick abundance between selected transects, considering factors such as plant cover seasonality, temperature, and humidity. Results: During the collection periods, we gathered 586 live ticks, with 20% found in grazed areas and 80% in ungrazed areas. Notably, tick abundance was significantly higher in ungrazed areas. Peaks in tick abundance occurred in both grazed and ungrazed areas during spring, particularly in April. However, tick numbers declined more rapidly in grazed areas. Microclimatic variables like temperature and humidity did not significantly impact tick abundance compared to landscape management and seasonal factors. Conclusion: Rewilding efforts, particularly natural grazing by large ungulates, influence tick abundance and distribution. This study provides empirical data on tick ecology in rewilded areas, highlighting the importance of landscape management and environmental factors in tick management and conservation. Trophic rewilding plays a crucial role in shaping ecosystems and tick population dynamics in transformed landscapes.

Borrelia spirochetes in European exotic farm animals.

Ticks transmit a broad spectrum of pathogens, threatening both animal and human health. Tick survival and proliferation are strongly dependent on host selection and suitability. The hard tick Ixodes ricinus, which is widespread throughout most of Europe, is a host generalist capable of feeding on many different vertebrate species. Pasture-kept exotic farm animals may be at a high risk for tick and tick-borne pathogens infestations but research characterizing this is currently lacking. This study focused on the detection of Borrelia spirochetes (including Borrelia miyamotoi) in exotic farm animals. Using nested-PCR with Borrelia-specific primers, 121 serum samples from 54 exotic farm animals of several species bred in four different farms in Bohemia and Moravia (Czechia) were tested. Positive samples were sequenced for the identification of Borrelia species. The prevalence of Borrelia DNA in the samples ranged from 13 to 67%, depending on the sampling site. The sequencing results confirmed the DNA presence of multiple spirochete species from the Borrelia burgdorferi sensu lato complex. Only one sample from an ostrich (Struthio camelus) was found to be positive for Borrelia myiamotoi. The results show that exotic farm animals can serve as hosts for hard ticks and can be infected by Borrelia spirochetes, transmitted by hard ticks. Therefore, these animals could play a relevant role in maintaining Borrelia spirochetes in nature.

Role of Zoo-Housed Animals in the Ecology of Ticks and Tick-Borne Pathogens-A Review.

Ticks are ubiquitous ectoparasites, feeding on representatives of all classes of terrestrial vertebrates and transmitting numerous pathogens of high human and veterinary medical importance. Exotic animals kept in zoological gardens, ranches, wildlife parks or farms may play an important role in the ecology of ticks and tick-borne pathogens (TBPs), as they may serve as hosts for local tick species. Moreover, they can develop diseases of varying severity after being infected by TBPs, and theoretically, can thus serve as reservoirs, thereby further propagating TBPs in local ecosystems. The definite role of these animals in the tick-host-pathogen network remains poorly investigated. This review provides a summary of the information currently available regarding ticks and TBPs in connection to captive local and exotic wildlife, with an emphasis on zoo-housed species.

Detection of Borrelia burgdorferi sensu stricto and Borrelia garinii DNAs in patient with Hyperkeratosis lenticularis perstans (Flegel disease).

Determination of the causative agent of erythema-like skin lesions in case of nonspecific superficial perivascular dermatitis was supported by histological examination and led to the latter diagnosis of Hyperkeratosis lenticularis perstans (Flegel disease) in patient. The presence of antibodies against Borrelia burgdorferi in patient serum was confirmed by a routine ELISA method and verified by Western blot technique. Skin biopsy and blood specimens were analyzed by PCR and multilocus sequence analysis (MLSA). Western blot method revealed IgG antibody response against two specific antigens, 17 and 83 kDa proteins. The recombinant test detected IgG antibody response against p100 and p41 antigens. The sequence analysis of amplicons from the selected genomic loci obtained from skin biopsy and serum samples revealed the presence of two species from B. burgdorferi sensu lato complex as a co-infection in this patient-B. burgdorferi sensu stricto (s.s.) and Borrelia garinii.

Seroprevalence of Borrelia burgdorferi sensu lato and tick-borne encephalitis virus in zoo animal species in the Czech Republic.

This study was conducted to evaluate the prevalence of antibodies against Borrelia bugdorferi (Bb) s.l. and tick-borne encephalitis virus (TBEV) in zoo animals in the Czech Republic. We collected 133 serum samples from 69 animal species from 5 zoos located in different parts of the country. The samples were obtained from even-toed ungulates (n=78; 42 species), odd-toed ungulates (n=32; 11 species), carnivores (n=13; 9 species), primates (n=2, 2 species), birds (n=3; 2 species), and reptiles (n=5; 3 species). A high antibody prevalence (60%) was observed for Bb s.l. On the other hand, only two animals had TBEV-specific antibodies: a markhor (Capra falconeri) and a reindeer (Rangifer tarandus), both from the same zoo, located in an area endemic for TBEV. Both of these animals were also positive for Bb s.l. antibodies. Our results indicate that a high number of animal species in the Czech zoos were exposed to Bb s.l. and that TBEV infection occurred at least in one of the investigated zoos. Considering the pathogenic potential of these two tick-borne pathogens, clinical and serological monitoring should be continued, and therapeutic and preventive measures should be taken when necessary.

Functional genomics studies of Rhipicephalus (Boophilus) annulatus ticks in response to infection with the cattle protozoan parasite, Babesia bigemina.

Ticks are obligate haematophagous ectoparasites of wild and domestic animals as well as humans, considered to be second worldwide to mosquitoes as vectors of human diseases, but the most important vectors of disease-causing pathogens in domestic and wild animals. Babesia spp. are tick-borne pathogens that cause a disease called babesiosis in a wide range of animals and in humans. In particular, Babesia bovis and Babesia bigemina are transmitted by cattle ticks, Rhipicephalus (Boophilus) annulatus and Rhipicephalus microplus, which are considered the most important cattle ectoparasites with major economic impacts on cattle production. The objectives of this study were to identify R. annulatus genes differentially expressed in response to infection with B. bigemina. Functional analyses were conducted on selected genes by RNA interference in both R. annulatus and R. microplus ticks. Eight hundred randomly selected suppression-subtractive hybridisation library clones were sequenced and analysed. Molecular function Gene Ontology assignments showed that the obtained tick sequences encoded for proteins with different cellular functions. Differentially expressed genes with putative functions in tick-pathogen interactions were selected for validation of SSH results by real-time reverse transcription-PCR. Genes encoding for TROSPA, calreticulin, ricinusin and serum amyloid A were over-expressed in B. bigemina-infected ticks while Kunitz-type protease inhibitor 5 mRNA levels were down-regulated in infected ticks. Functional analysis of differentially expressed genes by double stranded RNA-mediated RNAi showed that under the conditions of the present study knockdown of TROSPA and serum amyloid A significantly reduced B. bigemina infection levels in R. annulatus while in R. microplus, knockdown of TROSPA, serum amyloid A and calreticulin also reduced pathogen infection levels when compared with controls. Several studies have characterised the tick-pathogen interface at the molecular level. However, to our knowledge this is the first report of functional genomics studies in R. annulatus infected with B. bigemina. The results reported here increase our understanding of the role of tick genes in Babesia infection/multiplication.

New defensins from hard and soft ticks: similarities, differences, and phylogenetic analyses.

Despite the importance of ticks as vectors of disease very little is known about their immune system. Antimicrobial peptides, including defensins (phylogenetically ancient antibacterial peptides) are major components of innate immunity in ticks that have been shown to provide protection against gram-negative and gram-positive bacteria, fungi, viruses and protozoan parasites. With the aim of studying the evolution of the genes involved in tick defense, we identified the preprodefensin genes from four Ornithodoros tick species (O. papillipes: isoforms A, B, and D; O. tartakovskyi and O. puertoricensis: isoforms A and B; O. rostratus: isoform A) and from two Dermacentor tick species (D. reticulatus and D. marginatus: one isoform) not previously described. Phylogenetic analyses revealed that Ornithodoros defensin isoforms (A, B, C, and D) form 4 separate clades, while hard tick defensins are divided into several branches based on particular tick species.

Integration of a tick-borne encephalitis virus and Borrelia burgdorferi sensu lato into mountain ecosystems, following a shift in the altitudinal limit of distribution of their vector, Ixodes ricinus (Krkonose mountains, Czech Republic).

The altitudinal shift in the limit of Ixodes ricinus occurrence above the previously established altitude of 750 m above sea level has been monitored over the long-term (2002-2008) in the Krkonose Mts. (Giant Mts.), the highest in the Czech Republic, along two vertical transects in their eastern and central parts (600-1020 and 600-1270 m). Ticks were collected by flagging three times annually, and examined individually by PCR or RT-PCR for the presence of Borrelia burgdorferi sensu lato or tick-borne encephalitis virus (TBEV). A total of 5999 I. ricinus ticks were tested. TBEV RNA was detected in 26 ticks at up to 1140 m. Demonstration of TBEV in two larvae of I. ricinus indicates transovarial transmission. Similar infection rates in larvae and nymphs show vertical transmission in TBEV circulation to be very important under these mountain conditions. B. burgdorferi sensu stricto was found at up to 1040-1065 m, Borrelia garinii and Borrelia afzelii up to 1080-1140 m, and Borrelia valaisiana up to 1270 m. The total infection rates of nymphs and larvae were 7.3% and 2%, respectively. B. garinii was the most prevalent (37%), followed by B. afzelii (29%), B. burgdorferi s.s. (11%), and B. valaisiana (9%). Double to quadruple coinfections were detected in 32% of the infected ticks, most frequently B. garinii/B. afzelii. Predominance of B. garinii and B. valaisiana over B. afzelii suggests that small passerine birds moving on the ground are responsible for permanent local populations of I. ricinus in mountain localities with low numbers of small terrestrial mammals. The detection of B. burgdorferi sensu lato and TBEV in host-seeking larvae indicates an autochthonic infection. Upon analysis of the local climate we consider climate warming to be responsible for the spreading of ticks and tick-transmitted pathogens to higher altitudes.

Ticks (Ixodidae) from passerine birds in the Carpathian region.

Birds have been found to be a reservoir host of borrelia. In order to assess the situation in Slovakia ticks were collected from a total of 3057 mist-netted, ringed and released passerine birds in two locations at 500 m (in 2001) and 1000 m (in 2003) above sea level in the Bukovské Vrchy Hills, part of the Carpathian region in the north-east of Slovakia. A total of 75 birds of 16 species were infested with subadult ticks of Ixodes ricinus species (prevalence of parasitization 5%). Sixty-two larvae from 31 birds of 9 species and 80 nymphs from 52 birds of 15 species were found. The highest intensity of parasitization was observed on blackbirds Turdus merula, song thrushes T. philomelos and dunnocks Prunella modularis. Six Ixodes ricinus adult ticks were found on humans working with birds, and one I. ricinus female tick on their dog. In ticks, the presence of Rickettsia sp., Coxiella burnetii, Borrelia burgdorferi sensu lato and members of the Anaplasmataceae and Piroplasmidae, were investigated by polymerase chain reaction, followed by sequence analysis. Rickettsia sp. was found in 1 nymph from the European robin Erithacus rubecula, in 3 adult ticks (1 male, 2 females) from humans and in the tick from the dog. The closely related Ehrlichia- like species "Schotti variant" was detected in 1 nymph from the song thrush. Borrelia afzelii was identified in 1 male and B. garinii in 1 female tick collected on humans. Ixodes ricinus was found to be the vector of a wide spectrum of tick-borne pathogens in a mountainous area of the Carpathians. Because of the low yield of ticks and pathogens the importance of birds as reservoir hosts is still poorly understood.