Ricinus Larvae Research Articles

Long term evaluation of factors influencing the association of ixodid ticks with birds in Central Europe, Hungary

Birds play a crucial role in disseminating ticks that carry pathogens of high veterinary-medical importance. The aim of this study was to analyze data of a long-term tick collection from birds at a single stop-over site in Central Europe, Hungary. Over eight years (2015–2022) 5833 ticks (ten species) were collected from 2395 tick-infested birds. The most abundant species were Ixodes ricinus (n = 3971) and Haemaphysalis concinna (n = 1706). Ixodes ricinus nymphs and larvae were the most frequently occurring on resident and short-distance migratory birds with forest habitat but Ha. concinna was the most abundant species on reed-associated, long-distance migrants. Haemaphysalis concinna occurred mostly on birds feeding above the ground level, while I. ricinus predominated on ground feeding birds. Infestation with I. ricinus nymphs always peaked in the first half of the year, in contrast to larvae which were more abundant on avian hosts in the autumn. At the same time, Ha. concinna larvae and nymphs had their peak numbers in the summer. This is the first long-term study on the tick infestation of birds in Central Europe. The study shows that, migration distance, habitat type, and typical feeding level of birds, as well as characteristics of tick life cycle are all key factors in the role of birds as tick disseminators. It was revealed that Savi’s Warbler (Locustella luscinioides) is the most frequent hosts of Ha. concinna in Central Hungary.

Molecular Detection of Borrelia burgdorferi s.l. (Borreliella) and Chlamydia-Like Organism DNA in Early Developmental Stages of Arthropod Vector Species

Borrelia burgdorferi sensu lato (Bbsl) is spirochetes transmitted by ticks and known to cause Lyme disease. Chlamydia-like organisms (CLOs) comprise a large group of bacteria that can lead to serious health disorders, including miscarriage. Recently, CLOs have been found in ticks and patient skin biopsies. Due to the involvement of multiple potential vectors in the spread of these pathogens, the objective of this study was to confirm the presence of both organisms in the early developmental stages of selected vectors. Three potential vectors, Ixodes ricinus larvae, Culex pipiens larvae, and winged (unfed) adults of Lipoptena cervi, were collected in the Czech Republic in years 2019–2020. The presence of Bbsl and panchlamydial DNA was detected by PCR and positive samples were further analyzed by Sanger sequencing and phylogenetic tree construction. Bbsl DNA was proved in 1.5% (2/137) of I. ricinus larvae (identified as Borrelia afzelii and Borrelia garinii), in 1.7% (2/119) of C. pipiens larvae (both identified as B. garinii), and in 11% (3/27) of L. cervi (all identified as B. garinii). CLOs were identified in 0.7% (1/137) of I. ricinus larvae (Candidatus Protochlamydia) and in 7.4% (2/27) of L. cervi (unspecified genus), while C. pipiens larvae could not be evaluated (0%). This research represents the first investigation of the presence of CLOs in L. cervi. The detection of pathogen DNA in the early developmental stages of vectors suggests the potential for transgenerational transmission of Bbsl and CLOs in the selected vectors, although at a low rate.

Parasitic Arthropods of Soricinae Shrews in North-Eastern Poland.

The study of the ectoparasite fauna of the insectivores-Sorex araneus, Sorex minutus, Neomys fodiens, and Neomys anomalus (subfamily Soricinae)-was carried out in three locations in Poland: Białowieża National Park, Kosewo Górne in the Masurian Lake District, and in vicinity of Warsaw. Three species of Ixodidae ticks, eleven species of fleas, and four species of mites from the order Mesostigmata were noted. The most numerous ectoparasites are ticks Ixodes ricinus (larvae), Dermacentor reticulatus (nymphs), and fleas Palaeopsylla soricis, Megabothris walkeri, and Hystrichopsylla orientalis. These species show the highest prevalence and show the highest dominance index. The parasitofauna of S. araneus is much richer in species than other shrew species. The structure and dominance of parasite assemblages differ between locations.

Sand lizards (Lacerta agilis) decrease nymphal infection prevalence for tick‐borne pathogens Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum in a coastal dune ecosystem

Abstract Understanding which factors determine tick‐borne disease hazard can contribute to effective disease control. In Europe, the hazard of the pathogens Borrelia burgdorferi s.l. and Anaplasma phagocytophilum is determined by local tick densities (mainly Ixodes ricinus) and the reservoir competence of the host species community. Sand lizards (Lacerta agilis) are common hosts for larvae and nymphs of I. ricinus and non‐competent reservoirs for both pathogens. Consequently, high relative abundance of L. agilis is hypothesized to be associated with lower infection prevalence in nymphs. Here, we aimed to test whether this effectively occurs in natural settings. We sampled different habitat types within a heterogenous dune landscape at the Dutch coast and estimated (1) L. agilis densities, (2) host community competence, (3) the density and infection prevalence of questing I. ricinus ticks and (4) the number and infection prevalence of ticks feeding on L. agilis. Captured L. agilis had high tick burdens and contributed substantially to feeding I. ricinus larvae in their natural habitat. B. burgdorferi s.l. and A. phagocytophilum were virtually absent from feeding larvae and nymphs. The nymphal infection prevalence of both pathogens in questing ticks was lower in habitat types where L. agilis was more abundant. Hence, L. agilis strongly reduced community competence. The density of questing nymphs was higher in habitat types with denser vegetation and also varied more between habitat types than infection prevalence. As a result, nymphal density had a stronger effect on the density of infected ticks than did nymphal infection prevalence. Synthesis and applications. Coastal dune habitats favourable for L. agilis have lower densities of questing nymphs and a lower human infection hazard. These results might be applicable to similar ecosystems where L. agilis is present. From a public health perspective, this underlines the importance of preserving early successional habitat, as encroaching shrubs are associated with higher tick‐borne disease hazard, and vegetation removal might be a solution to reduce hazard in coastal dunes. The high degree of spatial heterogeneity in the abundance of tick‐borne pathogens also poses opportunities to manage recreational activities to limit human exposure to tick‐borne diseases.

Voles, shrews and red squirrels as sources of tick blood meals and tick-borne pathogens on an island in southwestern Finland

Molecular identification of the previous blood meal source of a questing tick (Acari: Ixodidae) from blood meal fragments was proposed a few decades ago. Following this, several blood meal assays have been developed and published, but none of them have been taken into widespread use. Recently, novel retrotransposon-based qPCR assays designed for detecting blood meal fragments of North American host species were published. We wanted to assess their function with host species present in Finland.Questing ticks were collected by cloth dragging in August-September 2021 from an island in southwestern Finland. DNA was extracted from Ixodes ricinus nymphs (n=438) and qPCR assays applied to identify larval blood meal sources (voles, shrews and red squirrels) and screen for several tick-borne human pathogens and other microbes with pathogenic potential [Borrelia spp. (including specific assays for Borrelia afzelii, Borrelia garinii, Borrelia valaisiana), Anaplasma phagocytophilum, Babesia spp., Rickettsia spp., and Neoehrlichia mikurensis].The probability of a nymph having fed as larva on either a vole, shrew or red squirrel was 0.34 (0.30 – 0.38; 95% confidence interval). Bacteria of the genus Borrelia were the most common pathogens detected, with host-specific probabilities of carrying Borrelia of 0.30 (0.18 – 0.44) for nymphs that had fed on voles, 0.23 (0.14 – 0.35) for nymphs that had fed on shrews, and 0.42 (0.28 – 0.58) for nymphs that had fed on red squirrels. Other microbes were rarely acquired from these hosts, apart from N. mikurensis from voles.This study highlights that shrews and red squirrels may equal voles as blood meal sources for I. ricinus larvae. Overall, variation in proportions of blood meals provided by these animals may be high across even proximate study areas. All studied host species appeared to be important sources for particularly Borrelia afzelii, and voles also for N. mikurensis.

Infestation of mice with ixodid ticks in forests of Belgorod region

Rodents are one of the largest groups of existing mammals and number more than 2,270 known species, which is almost 42 % of the total biodiversity of worldwide-distributed mammals (except Antarctica and some islands). They are well adapted to various habitats and are known to live alongside people and livestock complexes. The purpose of the study was to study infestation of mice with ixodid ticks in forests of the Belgorod region adjacent to livestock and poultry farms. The study was carried out in 2019-2022. The extensiveness of mice infestation was observed in Sylvaemus sylvaticus species, which was 13.26 % higher (P 0.05) than in Sylvimus flavicollis , and 24.33 % higher (P 0.01) than in Apodemus agrarius . The intensity of infestation with preimaginal (larvae and nymphs) stages of Ixodes ricinus ticks had wide fluctuations depending on the type of synanthropic rodent. Thus, the highest intensity was noted in Sylvaemus sylvaticus mice, which was 8.73 and 13.56 % higher (P 0.001) compared to Sylvimus flavicollis and Apodemus agrarius mice, respectively. The maximum infestation of male synanthropic mouse species was 31 ticks in Sylvaemus sylvaticus , 19 ticks in Sylvimus flavicollis , and 9 ticks in Apodemus agrarius . The intensity of infestation in synanthropic mice was the highest in the summer months (from July to August) and decreased slightly in autumn. In autumn, 41.2 % of the examined rodents were infested with ticks, the predominant species was Ixodes ricinus (larvae). Larvae and nymphs of Ixodes ricinus ticks were more often attached to rodents in the head area (mainly on the ears) - 82 %, less often on other parts of the body - 1 8 %. A smaller number of ticks were present on neck, body, legs, sometimes parasites were found on tail.

First record of Ixodes ricinus (Acari; Ixodidae) in European glass lizard (Pseudopus apodus; Anguidae) and a review of ectoparasite studies in reptiles in Turkey

Three adult European glass lizards (Pseudopus apodus Klembara) were caught in the garden of Faculty of Veterinary Medicine, Ondokuz Mayis University (Samsun, Turkey) and were observed to have a tick infestation. During macroscopic examination, tick samples were collected from the lateral groove on both sides of the lizard by using blunt-tip tweezers. All the samples were examined under the microscope and belonged to Ixodes ricinus (Linnaeus) larvae and nymphs. This is the first report of a tick species or infestation in European glass lizard. In addition, we provide a detailed review of all studies conducted to date on reptile ectoparasites in Turkey.

Evaluation of Vector Competence of Ixodes Ticks for Kemerovo Virus

Tick-borne viruses are responsible for various symptoms in humans and animals, ranging from simple fever to neurological disorders or haemorrhagic fevers. The Kemerovo virus (KEMV) is a tick-borne orbivirus, and it has been suspected to be responsible for human encephalitis cases in Russia and central Europe. It has been isolated from Ixodes persulcatus and Ixodes ricinus ticks. In a previous study, we assessed the vector competence of I. ricinus larvae from Slovakia for KEMV, using an artificial feeding system. In the current study, we used the same system to infect different tick population/species, including I. ricinus larvae from France and nymphs from Slovakia, and I. persulcatus larvae from Russia. We successfully confirmed the first two criteria of vector competence, namely, virus acquisition and trans-stadial transmission, for both tick species that we tested. The estimated infection rates of engorged and moulted ticks suggest specificities between viral strains and tick species/developmental stages.

Host in reserve: The role of common shrews (Sorex araneus) as a supplementary source of tick hosts in small mammal communities influenced by rodent population cycles.

Rodents often act as important hosts for ticks and as pathogen reservoirs. At northern latitudes, rodents often undergo multi‐annual population cycles, and the periodic absence of certain hosts may inhibit the survival and recruitment of ticks. We investigated the potential role of common shrews (Sorex araneus) to serve as a supplementary host source to immature life stages (larvae and nymphs) of a generalist tick Ixodes ricinus and a small mammal specialist tick I. trianguliceps, during decreasing abundances of bank voles (Myodes glareolus). We used generalized mixed models to test whether ticks would have a propensity to parasitize a certain host species dependent on host population size and host population composition across two high‐latitude gradients in southern Norway, by comparing tick burdens on trapped animals. Host population size was defined as the total number of captured animals and host population composition as the proportion of voles to shrews. We found that a larger proportion of voles in the host population favored the parasitism of voles by I. ricinus larvae (estimate = −1.923, p = .039) but not by nymphs (estimate = −0.307, p = .772). I. trianguliceps larvae did not show a lower propensity to parasitize voles, regardless of host population composition (estimate = 0.875, p = .180), while nymphs parasitized shrews significantly more as vole abundance increased (estimate = 2.106, p = .002). These results indicate that common shrews may have the potential to act as a replacement host during periods of low rodent availability, but long‐term observations encompassing complete rodent cycles may determine whether shrews are able to maintain tick range expansion despite low rodent availability.

Infection with Borrelia afzelii and manipulation of the egg surface microbiota have no effect on the fitness of immature Ixodes ricinus ticks

Arthropod vectors carry vector-borne pathogens that cause infectious disease in vertebrate hosts, and arthropod-associated microbiota, which consists of non-pathogenic microorganisms. Vector-borne pathogens and the microbiota can both influence the fitness of their arthropod vectors, and hence the epidemiology of vector-borne diseases. The bacterium Borrelia afzelii, which causes Lyme borreliosis in Europe, is transmitted among vertebrate reservoir hosts by Ixodes ricinus ticks, which also harbour a diverse microbiota of non-pathogenic bacteria. The purpose of this controlled study was to test whether B. afzelii and the tick-associated microbiota influence the fitness of I. ricinus. Eggs obtained from field-collected adult female ticks were surface sterilized (with bleach and ethanol), which reduced the abundance of the bacterial microbiota in the hatched I. ricinus larvae by 28-fold compared to larvae that hatched from control eggs washed with water. The dysbiosed and control larvae were subsequently fed on B. afzelii-infected or uninfected control mice, and the engorged larvae were left to moult into nymphs under laboratory conditions. I. ricinus larvae that fed on B. afzelii-infected mice had a significantly faster larva-to-nymph moulting time compared to larvae that fed on uninfected control mice, but the effect was small (2.4% reduction) and unlikely to be biologically significant. We found no evidence that B. afzelii infection or reduction of the larval microbiota influenced the four other life history traits of the immature I. ricinus ticks, which included engorged larval weight, unfed nymphal weight, larva-to-nymph moulting success, and immature tick survival. A retrospective power analysis found that our sampling effort had sufficient power (> 80%) to detect small effects (differences of 5% to 10%) of our treatments. Under the environmental conditions of this study, we conclude that B. afzelii and the egg surface microbiota had no meaningful effects on tick fitness and hence on the R0 of Lyme borreliosis.

Three Babesia species in Ixodes ricinus ticks from migratory birds in Sweden

BackgroundMigratory birds can cross geographical and environmental barriers and are thereby able to facilitate transmission of tick-borne pathogens both as carriers of infected ticks and as reservoirs of pathogenic microorganisms. Ixodes ricinus is one of the most abundant tick species in the Northern Hemisphere and a main vector of several Babesia species, some which pose a potential threat to human and animal health. At present only two cases of overt babesiosis in humans have so far been reported in Sweden. To better understand the potential role of birds as disseminators of zoonotic Babesia protozoan parasites, we investigated the presence of Babesia species in ticks removed from migratory birds.MethodsTicks were collected from birds captured at Ottenby Bird Observatory, south-eastern Sweden, from March to November 2009. Ticks were molecularly identified to species, and morphologically to developmental stage, and the presence of Babesia protozoan parasites was determined by real-time PCR.ResultsIn total, 4601 migratory birds of 65 species were examined for tick infestation. Ticks removed from these birds have previously been investigated for the presence of Borrelia bacteria and the tick-borne encephalitis virus. In the present study, a total of 1102 ticks were available for molecular analysis of Babesia protozoan parasites. We found that 2.4% of the ticks examined, all I. ricinus, were positive for mammal-associated Babesia species. Out of all Babesia-positive samples, Babesia venatorum was the most prevalent (58%) species, followed by Babesia microti (38%) and Babesia capreoli (4.0%). B. venatorum and B. capreoli were detected in I. ricinus larvae, whereas B. microti was only present in I. ricinus nymphs. This supports the view that the two first-mentioned species are vertically (transovarially) transmitted in the tick population, in contrast to B. microti. The largest number of Babesia-infected ticks was removed from the common redstart (Phoenicurus phoenicurus) and European robin (Erithacus rubecula).ConclusionsThis study reveals that Babesia protozoan parasites are present in ticks infesting migratory birds in south-eastern Sweden, which could potentially lead to the dissemination of these tick-borne microorganisms into new areas, thus posing a threat to humans and other mammals.Graphical

Red and fallow deer determine the density of Ixodes ricinus nymphs containing Anaplasma phagocytophilum

BackgroundThe density of Ixodes ricinus nymphs infected with Anaplasma phagocytophilum is one of the parameters that determines the risk for humans and domesticated animals to contract anaplasmosis. For this, I. ricinus larvae need to take a bloodmeal from free-ranging ungulates, which are competent hosts for A. phagocytophilum.MethodsHere, we compared the contribution of four free-ranging ungulate species, red deer (Cervus elaphus), fallow deer (Dama dama), roe deer (Capreolus capreolus), and wild boar (Sus scrofa), to A. phagocytophilum infections in nymphs. We used a combination of camera and live trapping to quantify the relative availability of vertebrate hosts to questing ticks in 19 Dutch forest sites. Additionally, we collected questing I. ricinus nymphs and tested these for the presence of A. phagocytophilum. Furthermore, we explored two potential mechanisms that could explain differences between species: (i) differences in larval burden, which we based on data from published studies, and (ii) differences in associations with other, non-competent hosts.ResultsPrincipal component analysis indicated that the density of A. phagocytophilum-infected nymphs (DIN) was higher in forest sites with high availability of red and fallow deer, and to a lesser degree roe deer. Initial results suggest that these differences are not a result of differences in larval burden, but rather differences in associations with other species or other ecological factors.ConclusionsThese results indicate that the risk for contracting anaplasmosis in The Netherlands is likely highest in the few areas where red and fallow deer are present. Future studies are needed to explore the mechanisms behind this association.Graphical abstract

Failed Disruption of Tick Feeding, Viability, and Molting after Immunization of Mice and Sheep with Recombinant Ixodes ricinus Salivary Proteins IrSPI and IrLip1.

To identify potential vaccine candidates against Ixodes ricinus and tick-borne pathogen transmission, we have previously sequenced the salivary gland transcriptomes of female ticks infected or not with Bartonella henselae. The hypothesized potential of both IrSPI (I. ricinus serine protease inhibitor) and IrLip1 (I. ricinus lipocalin 1) as protective antigens decreasing tick feeding and/or the transmission of tick-borne pathogens was based on their presumed involvement in dampening the host immune response to tick feeding. Vaccine endpoints included tick larval and nymphal mortality, feeding, and molting in mice and sheep. Whether the antigens were administered individually or in combination, the vaccination of mice or sheep elicited a potent antigen-specific antibody response. However, and contrary to our expectations, vaccination failed to afford protection against the infestation of mice and sheep by I. ricinus nymphs and larvae, respectively. Rather, vaccination with IrSPI and IrLip1 appeared to enhance tick engorgement and molting and decrease tick mortality. To the best of our knowledge, these observations represent the first report of induction of vaccine-mediated enhancement in relation to anti-tick vaccination.

Tripartite Interactions among Ixodiphagus hookeri, Ixodes ricinus and Deer: Differential Interference with Transmission Cycles of Tick-Borne Pathogens

For the development of sustainable control of tick-borne diseases, insight is needed in biological factors that affect tick populations. Here, the ecological interactions among Ixodiphagus hookeri, Ixodes ricinus, and two vertebrate species groups were investigated in relation to their effects on tick-borne disease risk. In 1129 questing ticks, I. hookeri DNA was detected more often in I. ricinus nymphs (4.4%) than in larvae (0.5%) and not in adults. Therefore, we determined the infestation rate of I. hookeri in nymphs from 19 forest sites, where vertebrate, tick, and tick-borne pathogen communities had been previously quantified. We found higher than expected co-occurrence rates of I. hookeri with deer-associated Anaplasma phagocytophilum, and lower than expected rates with rodent-associated Borrelia afzelii and Neoehrlichia mikurensis. The prevalence of I. hookeri in nymphs varied between 0% and 16% and was positively correlated with the encounter probability of ungulates and the densities of all life stages of I. ricinus. Lastly, we investigated the emergence of I. hookeri from artificially fed, field-collected nymphs. Adult wasps emerged from seven of the 172 fed nymphs. From these observations, we inferred that I. hookeri is parasitizing I. ricinus larvae that are feeding on deer, rather than on rodents or in the vegetation. Since I. hookeri populations depend on deer abundance, the main propagation host of I. ricinus, these wasps have no apparent effect on tick populations. The presence of I. hookeri may directly interfere with the transmission cycle of A. phagocytophilum, but not with that of B. afzelii or N. mikurensis.

Transovarial transmission of Borrelia spp., Rickettsia spp. and Anaplasma phagocytophilum in Ixodes ricinus under field conditions extrapolated from DNA detection in questing larvae

BackgroundIxodes ricinus constitutes the main European vector tick for the Lyme borreliosis pathogen Borrelia burgdorferi (sensu lato), the relapsing fever borrelia Borrelia miyamotoi, as well as Anaplasma phagocytophilum and several Rickettsia species. Under laboratory conditions, a transovarial transmission to the next tick generation is described for Rickettsia spp. and Borrelia spp., especially regarding B. miyamotoi, whereas the efficiency of transovarial transfer under field conditions is largely unstudied.MethodsIn order to better estimate the potential infection risk by tick larvae for humans and animals, 1500 I. ricinus larvae from 50 collected “nests” (larvae adhering to the flag in a clumped manner) were individually examined for Borrelia, Rickettsia and A. phagocytophilum DNA using quantitative real-time PCR (qPCR).ResultsThirty-nine of 50 nests each (78.0%, 95% CI: 64.0–88.5%) were positive for Borrelia spp. and Rickettsia spp. DNA, and in three nests (6.0%, 95% CI: 1.3–16.5%) A. phagocytophilum DNA was detected. Overall, DNA from at least one pathogen could be detected in 90.0% (45/50, 95% CI: 78.2–96.7%) of the nests. Of the 1500 larvae, 137 were positive for Borrelia spp. DNA (9.1%, 95% CI: 7.7–10.7%), 341 for Rickettsia spp. DNA (22.7%, 95% CI: 20.6–24.9%) and three for A. phagocytophilum DNA (0.2%, 95% CI: 0–0.6%). Quantity of Borrelia spp. and Anaplasma spp. DNA in positive larvae was low, with 2.7 × 100Borrelia 5S-23S gene copies and 2.4 × 101A. phagocytophilum msp2/p44 gene copies detected on average, while Rickettsia-positive samples contained on average 5.4 × 102gltA gene copies. Coinfections were found in 66.0% (33/50, 95% CI: 51.2–78.8%) of the nests and 8.6% (38/443, 95% CI: 6.1–11.6%) of positive larvae. In fact, larvae had a significantly higher probability of being infected with Borrelia spp. or Rickettsia spp. when both pathogens were present in the nest.ConclusionsThis study provides evidence for transovarial transmission of Rickettsia spp. and Borrelia spp. in I. ricinus under field conditions, possibly facilitating pathogen persistence in the ecosystem and reducing the dependence on the presence of suitable reservoir hosts. Further studies are needed to prove transovarial transmission and to explain the surprisingly high proportion of nests containing Rickettsia and/or Borrelia DNA-positive larvae compared to infection rates in adult ticks commonly reported in other studies.

Research paper on abiotic factors and their influence on Ixodes ricinus activity\u2014observations over a two-year period at several tick collection sites in Germany

Tick-borne diseases are a public health issue. To predict vector tick abundance and activity, it is necessary to understand the driving factors for these variables. In this study, the activity of Ixodes ricinus was investigated in forest and meadow habitats in Germany with a focus on abiotic factors. Ixodes ricinus adults, nymphs and larvae were caught by flagging over a period of 2 years. Microclimatic and weather conditions were recorded at the collection sites. Statistical models were applied to describe correlations between abiotic factors and tick activity in univariable and multivariable analyses. Tick activity was observed in a broad range of air temperature between 3 and 28 °C, and air humidity varied between 35 and 95%. In general, tick activity of nymphs and larvae was higher in forest habitats than that in meadows. With the exception of a single specimen of Dermacentor reticulatus, all ticks were Ixodes ricinus, most of them nymphs (63.2% in 2009 and 75.2% in 2010). For the latter, a negative binomial mixed-effects model fitted best to the observed parameters. The modelling results showed an activity optimum between 20 and 23 °C for air temperature and between 13 and 15 °C for ground temperature. In univariable analyses, the collection site, month, season, ground and air temperature were significant factors for the number of ticks caught and for all life stages. In the multivariable analysis, temperature, season and habitat turned out to be key drivers. Ixodes ricinus positive for RNA of tick-borne encephalitis virus was only found at a single sampling site. The results of this study can be used in risk assessments and to parameterise predictive models.

Effect of rodent density on tick and tick-borne pathogen populations: consequences for infectious disease risk

BackgroundRodents are considered to contribute strongly to the risk of tick-borne diseases by feeding Ixodes ricinus larvae and by acting as amplifying hosts for pathogens. Here, we tested to what extent these two processes depend on rodent density, and for which pathogen species rodents synergistically contribute to the local disease risk, i.e. the density of infected nymphs (DIN).MethodsIn a natural woodland, we manipulated rodent densities in plots of 2500 m2 by either supplementing a critical food source (acorns) or by removing rodents during two years. Untreated plots were used as controls. Collected nymphs and rodent ear biopsies were tested for the presence of seven tick-borne microorganisms. Linear models were used to capture associations between rodents, nymphs, and pathogens.ResultsInvestigation of data from all plots, irrespective of the treatment, revealed a strong positive association between rodent density and nymphal density, nymphal infection prevalence (NIP) with Borrelia afzelii and Neoehrlichia mikurensis, and hence DIN’s of these pathogens in the following year. The NIP, but not the DIN, of the bird-associated Borrelia garinii, decreased with increasing rodent density. The NIPs of Borrelia miyamotoi and Rickettsia helvetica were independent of rodent density, and increasing rodent density moderately increased the DINs. In addition, NIPs of Babesia microti and Spiroplasma ixodetis decreased with increasing rodent density, which had a non-linear association with DINs of these microorganisms.ConclusionsA positive density dependence for all rodent- and tick-associated tick-borne pathogens was found, despite the observation that some of them decreased in prevalence. The effects on the DINs were variable among microorganisms, more than likely due to contrasts in their biology (including transmission modes, host specificity and transmission efficiency). The strongest associations were found in rodent-associated pathogens that most heavily rely on horizontal transmission. Our results draw attention to the importance of considering transmission mode of a pathogen while developing preventative measures to successfully reduce the burden of disease.

The effect of sublethal concentrations of deltamethrin and alphacypermethrin on the fecundity and development of Ixodes ricinus (Acari: Ixodidae) eggs and larvae

Two pyrethroids, deltamethrin (D) and alphacypermethrin (AC), have been used as commercial products for tick control worldwide. However, the effects of sublethal doses of these compounds on various tick species and their developmental stages have not been fully explored yet, although such knowledge could contribute to a more effective application of both pyrethroids and simultaneous reduction of their costs and undesirable side effects, including the harmful impact on other organisms and environmental contamination. This study investigates the effect of sublethal concentrations of D and AC applied to engorged females on the fecundity, development of eggs and larvae, and the number of offspring in the Ixodes ricinus (L.) tick, which is the most important vector of tick-borne diseases of humans in Europe. After detachment from rabbit’s skin, fully engorged I. ricinus females were treated with 20 μl of pyrethroid solutions at five concentrations from 0.01562 to 0.25% and kept at 28 °C and 75% relative humidity. The impact of the pyrethroids on maturation and development of eggs as well as development of larvae was assessed based on parameters of the preoviposition and oviposition periods and the course of embryogenesis and egg hatch. The investigations have shown that both tested acaricides reduce the reproductive rate in I. ricinus females by inhibition of egg development and disturb embryonic development and larval hatch. Females did not lay eggs at concentrations higher than 0.0625% AC and 0.125% D. The lower concentrations of both pyrethroids disturbed or inhibited the embryogenesis and egg hatch in I. ricinus.

Repeated isolation of tick-borne encephalitis virus from adult Dermacentor reticulatus ticks in an endemic area in Germany

BackgroundTick-borne encephalitis (TBE) virus is transmitted to humans and animals through tick bites and is thought to circulate in very strictly defined natural environments called natural foci. The most common tick serving as a vector for the TBE virus in central Europe is Ixodes ricinus; it is rarely found in other tick species and in Dermacentor reticulatus it has, so far, only been reported in Poland.MethodsBetween autumn 2016 and spring 2018 ticks were collected by the flagging method in a new TBE focus in the district of northern Saxony, Germany, outside the known risk areas as defined by the national Robert Koch Institute. Ticks were morphologically identified and tested in pools for the presence of TBE virus using a real-time RT-PCR. TBE virus from positive pools was isolated in A549 cells, and the E gene sequences were determined after conventional RT-PCR, followed by a phylogenetic comparison.ResultsTBE virus was detected in 11 pools, 9 times in flagged adults D. reticulatus (n = 1534; MIR: 0.59%, CI: 0.29–11.3%) and only twice in I. ricinus nymphs (n = 349; MIR: 0.57%, CI: 0.02–2.2%). All other ticks, I. ricinus males (n = 33), females (n = 30) and larvae (n = 58), as well as 5 I. inopinatus (2 females, 3 males) and 14 Haemaphysalis concinna (3 females, 11 nymphs), tested negative for TBE virus. TBE virus was not detected in I. ricinus during the summer, when D. reticulatus is not active. Sequence comparison of the entire E gene of the isolated virus strains resembled each other with only 3 nucleotide differences. The most closely related viral sequences belong to TBE virus strains from Poland and Neustadt an der Waldnaab (county of Neustadt an der Waldnaab, Bavaria), approximately 200 km east and 200 km south-west of the new focus, respectively.ConclusionsTBE virus was found in northern Saxony, Germany, with similar MIRs in D. reticulatus and I. ricinus, indicating that D. reticulatus plays an equal role to I. ricinus in virus circulation when both tick species are sympatric.

Unravelling the phenology of Ixodes frontalis, a common but understudied tick species in Europe

Tick-borne diseases have a complex epidemiology that depends on different ecological communities, associating several species of vertebrate hosts, vectors and pathogens. While most studies in Europe are focused on Ixodes ricinus, the common sheep tick, other Ixodes species may also be involved in the transmission or maintenance of different pathogens. In this study, we describe for the first time the activity pattern of I. frontalis, an understudied but widespread tick species associated with several common bird species in Europe. Questing ixodid tick stages (larvae, nymphs and adults) of both I. frontalis and I. ricinus were monitored by the drag sampling method over three years at the same locations in Western France. Differential activities were observed depending on I. frontalis life stages: nymphs and adults were present sporadically on the ground throughout the year, while larvae exhibited a marked peak of activity around October-November with tens or even hundreds of individuals per m2, followed by a slow decrease in winter. Larvae were completely absent in summer, which contrasts with the high numbers of I. ricinus larvae at this time of the year. The vegetation and the litter where the two tick species were found also exhibited marked differences, with I. frontalis mostly collected under bamboo bushes.