Tick Stages Research Articles

Description of ticks species from Rattus tiomanicus in Mangrove Forests through scanning electron microscopy

Ticks are ill-famed vectors of many pathogenic organisms which can cause various diseases and life-threatening illnesses to animals and humans. Each tick’s species and its life stages have distinct morphological features that can permit them to be accurately identified. However, the use of conventional stereo microscopes limits the accuracy of species identification. The taxonomy of ticks, in general, is not much understood and existing information is based on sparse morphology data. Thus, this study aims to examine and describe the morphological characteristics of different species of ticks collected on rodents in mangrove forests using scanning electron microscopy (SEM). This method renders high-quality images of body parts of ticks. Five different morphospecies of ticks from one host species (Rattus tiomanicus) were examined under SEM, followed by the PCR technique using mitochondrial 16S rDNA gene for species validation. This study revealed that the ticks belong to five species: Dermacentor auratus, Ixodes granulatus, Haemaphysalis hystricis, D. atrosignatus and Amblyomma cordiferum. The combination of stereomicroscopic and SEM methods has improved our understanding of the morphological characteristics of different tick species, hence establishing up-to-date taxonomic keys for these species. Moreover, due to the lack of taxonomic keys on the immature stage of ticks, the SEM method is essential in characterising the morphological features of these stages in detail, subsequently helpful in revising the taxonomic keys for certain ticks species.

Ectoparasitism of castor bean ticks Ixodes ricinus (Linnaeus, 1758) on sand lizards Lacerta agilis (Linnaeus, 1758) in western Poland

Background. Over 700 species of ticks are characterised by moderate to high host specificity, and a handful by very high host specificity, parasitizing upon a single host species. The rest are not host specific. One of them is the castor bean tick, which parasitizes all available terrestrial vertebrates including humans. Poland lacks tick species that are reptile-specific, since, as reported by K. Siuda (1993), these ectoparasites generally prefer endotherms, namely birds and mammals. However, the literature reports that the prevalence of castor bean tick infestation of local sand lizard populations is as high as 13.2 % – 40.4 %. Observations of different authors also indicate that outside the period of regular tick appearance on lizards, individual lizards can have very high tick loads, varying between several to over 100. Materials and Methods. The research was conducted in three separate study areas in western Poland: Żary, Zielona Góra and Nowa Sól. 606 specimens of sand lizards were captured, analysed and photographed. 222 (36.6 %) of them were infested with ticks. With each capture ticks were checked and removed. The lizards were caught by hand. Inter-group two-way and three-way ANOVA and Pearson correlation coefficients were calculated with the use of IBM SPSS Statistics 23 software. Threshold of statistical significance was set to α = 0.05. Results. In total 1339 ticks were obtained from lizards, all of which were castor bean ticks (Ixodes ricinus). 914 (68.2 %) were larvae, 424 (31.7 %) were nymphs and 1 (0.1 %) was an adult female. The highest number of ticks (778) was collected from lizards at Żary site. The most infested male from Żary was parasitized upon by 55 ticks, from Zielona Góra – by 46 and from Nowa Sól – by 15. In regard to females these numbers were 36, 34 and 15, respectively. The average infestation (defined as number of ticks) was higher for lizards from Żary than for those from Zielona Góra (p < 0.001) and Nowa Sól (p < 0.001), which also differed from each other in a statistically significant way (p = 0.001). Main effect of sex/age group turned out to be statistically significant, F(2.796) = 67.75; p < 0.001; η2 = 0.15. Hence post-hoc analysis was carried out and showed that males carried higher number of ticks than females and juveniles (p < 0.001). Females, on the other hand, carried more ticks than juveniles (p < 0.001). Conclusion. The research confirmed a significant role of sand lizards in propagation of Ixodes ricinus. It was shown that lizards were affected mainly by early develop­mental stages of ticks. The prevalence of tick infestation was 36.6 % – 58.1 % for Żary, 28.1 % for Zielona Góra and 13.8 % for Nowa Sól. The average tick load on sand lizards was 2.5 individuals. The highest number of them was collected from 58.5 % of males, 36.4 % of females and 5.1 % of juveniles. High tick infestation of males can be explained by their higher mobility, especially during mating season.

Acaricidal Efficacy of Biosynthesized Zinc Oxide Nanoparticles Against Hyalomma dromedarii (Acari: Ixodidae) and Their Toxic Effects on Swiss Albino Mice

PurposeThe current study aimed to investigate the efficacy of zinc oxide nanoparticles (ZnO NPs) synthesized by Melia azedarach aqueous extract to control Hyalomma dromedarii tick, and to evaluate their toxic effects on Swiss albino mice.MethodsZnO NPs were synthesized using M. azedarach aqueous extract. UV–visible spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive spectroscopy were used to characterize the synthesized NPs. Egg, nymph, larva, and adult immersion tests were used for bioassay of tick stages with the synthesized ZnO NP. A toxicity study was performed on Swiss albino mice after treatment with 1/10 of the oral LD50 of ZnO NPs (8437 mg/kg) for 5 successive days by oral gavage.ResultsThe LC50 of ZnO NPs on the eggs, larvae, and nymphs was 11.6, 8.03, and 3.9 mg/ml, respectively. The reproductive performance of females treated with ZnO NPs was lower than that of untreated females. The hematological results showed an insignificant increase in the level of white blood cells with normal red blood cells, hemoglobin, in addition to normal platelet count. The biochemical analysis showed an insignificant increased level (P > 0.05) of alkaline phosphatase and alanine aminotransferase. The liver and kidney suffered few histopathological changes after oral administration of ZnO NPs.ConclusionThese results suggest that ZnO NPs have good acaricidal activity against eggs, larvae, and engorged nymphs of H. dromedarii. ZnO NPs minimized the number of eggs laid by engorged females and the hatchability of their eggs. ZnO NPs did not affect unfed adults. The toxicity results of the mice revealed insignificant changes in the hemogram, biochemistry, with liver and kidney suffering few histopathological changes. Future studies are needed to assess application routes (topical vs oral). Based on these findings, ZnO NPs may be incorporated in the control of camel tick H. dromedarii.

Context-dependent host dispersal and habitat fragmentation determine heterogeneity in infected tick burdens: an agent-based modelling study.

As the incidence of tick-borne diseases has sharply increased over the past decade, with serious consequences for human and animal health, there is a need to identify ecological drivers contributing to heterogeneity in tick-borne disease risk. In particular, the relative importance of animal host dispersal behaviour in its three context-dependent phases of emigration, transfer and settlement is relatively unexplored. We built a spatially explicit agent-based model to investigate how the host dispersal process, in concert with the tick and host demographic processes, habitat fragmentation and the pathogen transmission process, affects infected tick distributions among hosts. A sensitivity analysis explored the impacts of different input parameters on infected tick burdens on hosts and infected tick distributions among hosts. Our simulations indicate that ecological predictors of infected tick burdens differed among the post-egg life stages of ticks, with tick attachment and detachment, tick questing activity and pathogen transmission dynamics identified as key processes, in a coherent way. We also found that the type of host settlement strategy and the proportion of habitat suitable for hosts determined super-spreading of infected ticks. We developed a theoretical mechanistic framework that can serve as a first step towards applied studies of on-the-ground public health intervention strategies.

Molecular Characterization and Gene Expression Analysis of Aquaporin in Haemaphysalis qinghaiensis.

Aquaporins (AQPs) are important functional proteins and are widely present in the cell membrane of almost all organisms, mediating transmembrane transport of liquid and other solutes. Much is known about the molecular characterization of AQPs in other tick species; however, nothing is known about them in Haemaphysalis qinghaiensis. In this study, we first sequenced the transcript variants of AQPs in H. qinghaiensis (HqAQPs), analyzed the biological structure features of AQPs, and investigated the pattern of gene expression of the AQP gene of H. qinghaiensis in different tick tissues and stages to predict their biological functions. In conclusion, four AQP transcript variants (i.e., HqAQP1-1, HqAQP1-2, HqAQP1-3, and HqAQP1-4) of H. qinghaiensis were found, and the sequences were comparable with its orthologs from the reported tick species. Gene expression of AQPs in different tick tissues and stages showed the higher expression level in salivary glands and gut of adult female, as well as in the female and nymph than in Malpighian tubules, ovary, male, larvae, and egg. Further studies will be performed to evaluate the function of HqAQPs against H. qinghaiensis infestation on animals.

Current and future distribution of Ixodes scapularis ticks in Québec: Field validation of a predictive model.

The incidence of Lyme disease is increasing in Québec and is closely linked to the distribution of Ixodes scapularis ticks. A time-to-establishment model developed in 2012 by Leighton and colleagues predicted the year of tick population establishment for each municipality in eastern Canada. To validate if this model correctly predicted tick distribution in Québec, predicted tick establishment was compared to field data from active tick surveillance (2010-2018) using two criteria: i) the detection of at least one tick and ii) the detection of the three questing stages of the tick. The speed of tick establishment and the increase in the exposed human population by 2100 were predicted with the time-to-establishment model. Field observations were consistent with model predictions. Ticks were detected on average 3 years after the predicted year. The probability of tick detection is significantly higher after the predicted year than before (61% vs 27% of collections). The trend was similar for the detection of three tick stages (16% vs 9% of collections). The average speed of tick range expansion was estimated by the model to be 18 km/year in Québec, with 90% of the human population exposed by 2027. The validation of the time-to-establishment model using field data confirmed that it could be used to project I. scapularis range expansion in Québec, and consequently the increase in Lyme disease risk over the coming decades. This will help public health authorities anticipate and adapt preventive measures, especially in areas not yet affected by Lyme disease.

The Bacterial Community in Questing Ticks From Khao Yai National Park in Thailand.

Ticks are known vectors for a variety of pathogens including bacteria, viruses, fungi, and parasites. In this study, bacterial communities were investigated in active life stages of three tick genera (Haemaphysalis, Dermacentor, and Amblyomma) collected from Khao Yai National Park in Thailand. Four hundred and thirty-three questing ticks were selected for pathogen detection individually using real-time PCR assays, and 58 of these were subjected to further metagenomics analysis. A total of 62 ticks were found to be infected with pathogenic bacteria, for a 14.3% prevalence rate, with Amblyomma spp. exhibiting the highest infection rate (20.5%), followed by Haemaphysalis spp. (14.5%) and Dermacentor spp. (8.6%). Rickettsia spp. were the most prevalent bacteria (7.9%) found, followed by Ehrlichia spp. (3.2%), and Anaplasma spp. and Borrelia spp. each with a similar prevalence of 1.6%. Co-infection between pathogenic bacteria was only detected in three Haemaphysalis females, and all co-infections were between Rickettsia spp. and Anaplasmataceae (Ehrlichia spp. or Anaplasma spp.), accounting for 4.6% of infected ticks or 0.7% of all examined questing ticks. The prevalence of the Coxiella-like endosymbiont was also investigated. Of ticks tested, 65.8% were positive for the Coxiella-like endosymbiont, with the highest infection rate in nymphs (86.7%), followed by females (83.4%). Among tick genera, Haemaphysalis exhibited the highest prevalence of infection with the Coxiella-like endosymbiont. Ticks harboring the Coxiella-like endosymbiont were more likely to be infected with Ehrlichia spp. or Rickettsia spp. than those without, with statistical significance for Ehrlichia spp. infection in particular (p-values = 0.003 and 0.917 for Ehrlichia spp. and Rickettsia spp., respectively). Profiling the bacterial community in ticks using metagenomics revealed distinct, predominant bacterial taxa in tick genera. Alpha and beta diversities analyses showed that the bacterial community diversity and composition in Haemaphysalis spp. was significantly different from Amblyomma spp. However, when examining bacterial diversity among tick life stages (larva, nymph, and adult) in Haemaphysalis spp., no significant difference among life stages was detected. These results provide valuable information on the bacterial community composition and co-infection rates in questing ticks in Thailand, with implications for animal and human health.

Infection rates, species diversity, and distribution of zoonotic Babesia parasites in ticks: a global systematic review and meta-analysis.

Zoonotic Babesia species are emerging public health threats globally, and are the cause of a mild to severe malaria-like disease which may be life threatening in immunocompromised individuals. In this study, we determine the global infection rate, distribution, and the diversity of zoonotic Babesia species in tick vectors using a systematic review and meta-analysis. We used the random-effects model to pool data and determined quality of individual studies using the Joanna Briggs Institute critical appraisal instrument for prevalence studies, heterogeneity using Cochran's Q test, and across study bias using Egger's regression test. Herein, we reported a 2.16% (3915/175345, 95% CI: 1.76-2.66) global infection rate of zoonotic Babesia species (B. divergens, B. microti, and B. venatorum) in tick vectors across 36 countries and 4 continents. Sub-group infection rates ranged between 0.65% (95% CI: 0.09-4.49) and 3.70% (95% CI: 2.61-5.21). B. microti was the most prevalent (1.79%, 95% CI: 1.38-2.31) species reported in ticks, while Ixodes scapularis recorded the highest infection rate (3.92%, 95% CI: 2.55-5.99). Larvae 4.18% (95% CI: 2.15-7.97) and females 4.08% (95% CI: 2.56-6.43) were the tick stage and sex with the highest infection rates. The presence of B. divergens, B. microti, and B. venatorum in tick vectors as revealed by the present study suggests possible risk of transmission of these pathogens to humans, especially occupationally exposed population. The control of tick vectors through chemical and biological methods as well as the use of repellants and appropriate clothing by occupationally exposed population are suggested to curtail the epidemiologic, economic, and public health threats associated with this emerging public health crisis.

Recent Advances in Molecular Genetic Tools for Babesia.

Development of in vitro culture and completion of genome sequencing of several Babesia parasites promoted the efforts to establish transfection systems for these parasites to dissect the gene functions. It has been more than a decade since the establishment of first transfection for Babesia bovis, the causative agent of bovine babesiosis. However, the number of genes that were targeted by genetic tools in Babesia parasites is limited. This is partially due to the low efficiencies of these methods. The recent adaptation of CRISPR/Cas9 for genome editing of Babesia bovis can accelerate the efforts for dissecting this parasite’s genome and extend the knowledge on biological aspects of erythrocytic and tick stages of Babesia. Additionally, glmS ribozyme as a conditional knockdown system is available that could be used for the characterization of essential genes. The development of high throughput genetic tools is needed to dissect the function of multigene families, targeting several genes in a specific pathway, and finally genome-wide identification of essential genes to find novel drug targets. In this review, we summarized the current tools that are available for Babesia and the genes that are being targeted by these tools. This may draw a perspective for the future development of genetic tools and pave the way for the identification of novel drugs or vaccine targets.

Elevational Changes in Bacterial Microbiota Structure and Diversity in an Arthropod-Disease Vector

Environmental conditions change rapidly along elevational gradients and have been found to affect community composition in macroscopic taxa, with lower diversity typically observed at higher elevations. In contrast, microbial community responses to elevation are still poorly understood. Specifically, the effects of elevation on vector-associated microbiota have not been studied to date, even though the within-vector microbial community is known to influence vector competence for a range of zoonotic pathogens. Here we characterize the structure and diversity of the bacterial microbiota in an important zoonotic disease vector, the sheep tick Ixodes ricinus, along replicated elevational gradient (630–1673 m) in the Swiss Alps. 16S rRNA sequencing of the whole within-tick bacterial microbiota of questing nymphs and adults revealed a decrease in Faith’s phylogenetic microbial alpha diversity with increasing elevation, while beta diversity analyses revealed a lower variation in microbial community composition at higher elevations. We also found a higher microbial diversity later in the season and significant differences in microbial diversity among tick life stages and sexes, with lowest microbial alpha diversity observed in adult females. No associations between tick genetic diversity and bacterial diversity were observed. Our study demonstrates systematic changes in tick bacterial microbiota diversity along elevational gradients. The observed patterns mirror diversity changes along elevational gradients typically observed in macroscopic taxa, and they highlight the key role of environmental factors in shaping within-host microbial communities in ectotherms.

Molecular detection of Cercopithifilaria bainae in brown dog ticks collected from dogs across the United States

Cercopithifilaria bainae is a filarioid nematode of dogs shown to use Rhipicephalus sanguineus sensu lato (s.l.), the brown dog tick, as the vector. Previously in the United States, C. bainae infections have been reported in a dog from Florida, and in dogs and ticks in Oklahoma, but data are lacking from other areas of the country. Here, we tested brown dog ticks from across the United States for C. bainae DNA to assess the geographic distribution of where this novel parasite may be cycling in ticks and dogs. Archival brown dog ticks were available for testing through the national tick survey Show Us Your Ticks. Ticks were morphologically identified, dissected, and tested by PCR to detect filarioid mitochondrial DNA. A total of 1400 brown dog ticks were tested from 321 separate animals from 23 states, with 5.7 % (80/1400) of the ticks testing positive for C. bainae DNA. At least one positive tick was detected in submissions from 9 states in addition to Florida and Oklahoma. Cercopithifilaria bainae DNA was detected in larval, nymphal, and adult stages of brown dog ticks and only in ticks removed from dogs. Of all dogs with brown dog ticks collected from them, 17.6 % (55/312) were infested with at least one tick that harbored C. bainae DNA. Findings from this study demonstrate a wider geographic range of C. bainae than previously known, and that dogs are commonly infested with brown dog ticks with molecular evidence of C. bainae infection.

Fine-scale determinants of the spatiotemporal distribution of Ixodes scapularis in Quebec (Canada)

The tick vector of Lyme disease, Ixodes scapularis, is currently expanding its geographical distribution northward into southern Canada driving emergence of Lyme disease in the region. Despite large-scale studies that attributed different factors such as climate change and changes in land use to the geographical expansion of the tick, a comprehensive understanding of local patterns of tick abundance is still lacking in that region. Using a newly endemic periurban nature park located in Quebec (Canada) as a model, we explored intra-habitat patterns in tick distribution and their relationship with biotic and abiotic factors. We verified the hypotheses that (1) there is spatial heterogeneity in tick densities at the scale of the park and (2) these patterns can be explained by host availability, habitat characteristics and microclimatic conditions. During tick activity season in three consecutive years, tick, deer, rodent and bird abundance, as well as habitat characteristics and microclimatic conditions, were estimated at thirty-two sites. Patterns of tick distribution and abundance were investigated by spatial analysis. Generalised additive mixed models were constructed for each developmental stage of the tick and the relative importance of significant drivers on tick abundance were derived from final models. We found fine-scale spatial heterogeneity in densities of all tick stages across the park, with interannual variability in the location of hotspots. For all stages, the local density was related to the density of the previous stage in the previous season, in keeping with the tick's life cycle. Adult tick density was highest where drainage was moderate (neither waterlogged nor dry). Microclimatic conditions influenced the densities of immature ticks, through the effects of weather at the time of tick sampling (ambient temperature and relative humidity) and of the seasonal microclimate at the site level (degree-days and number of tick adverse moisture events). Seasonal phenology patterns were generally consistent with expected curves for the region, with exceptions in some years that may be attributable to founder events. This study highlights fine scale patterns of tick population dynamics thus providing fundamental knowledge in Lyme disease ecology and information applicable to the development of well-targeted prevention and control strategies for public natural areas affected by this growing problem in southern Canada.

Drop-off rhythms and survival of Hyalomma anatolicum (Acari: Ixodidae) fed on crossbred (Friesians x Zebu) calves in Sudan

Hyalomma anatolicum is one of the most economically important ticks in the Sudan. It is the main vector of tropical theileriosis in cattle and other diseases in different species. The study aims to investigate the drop-off rhythms and survival periods of tick stages fed on crossbred calves. Larvae, nymphs and adults fed on crossbred calves (male, 3–6 months old, Friesians x Zebu) kept under zero grazing system. Drop-off rhythms of engorged stages were studied under field conditions while the survival periods of unfed stages were investigated under field and laboratory conditions (27 °C and 85% R.H.). Significantly high numbers of engorged larvae dropped at night between 20H:00 and 07H:00. (March, April and August). Engorged nymphs dropped late evening and early night 16H:00–20H:00. (September), while insignificant number of engorged females dropped between 10H:00 and 14H:00 (October). Survival durations of unfed H. anatolicum stages were longer under laboratory conditions than in the field. These were 99.36 ± 1.24 and 13.12 ± 0.68 days for larvae, 63 ± 1.33 and 16 ± 0.87 days for nymphs and 90 ± 3.6 and 45 ± 2.7 days for adults, respectively. Under field condition survival studies were conducted in May, August and December 2016 for larvae, March, May, August and December 2016 for nymphs and in October 2016 for Adults. In August, 70% of unfed larvae and nymphs survived for three weeks and 2.5 weeks, respectively, in May, 16% of unfed larvae survived for one week while no unfed nymphs survived for more than 4 days. This finding may indicate that larvae are more resistant to desiccation than nymphs. The abnormal changes on the alloscutum of H. anatolicum desiccated nymphs and females were observed being shrunken and truncated posteriorly, a finding which opens new avenues for further studies. For control strategies more studies are recommended on the dropping to allow ticks drop in an area not suitable for further development as one of the control strategies and the survival studies also is exploited in rotational grazing of animals.

Lessons Learned for Pathogenesis, Immunology, and Disease of Erythrocytic Parasites: Plasmodium and Babesia.

Malaria caused by Plasmodium species and transmitted by Anopheles mosquitoes affects large human populations, while Ixodes ticks transmit Babesia species and cause babesiosis. Babesiosis in animals has been known as an economic drain, and human disease has also emerged as a serious healthcare problem in the last 20–30 years. There is limited literature available regarding pathogenesis, immunity, and disease caused by Babesia spp. with their genomes sequenced only in the last decade. Therefore, using previous studies on Plasmodium as the foundation, we have compared similarities and differences in the pathogenesis of Babesia and host immune responses. Sexual life cycles of these two hemoparasites in their respective vectors are quite similar. An adult Anopheles female can take blood meal several times in its life such that it can both acquire and transmit Plasmodia to hosts. Since each tick stage takes blood meal only once, transstadial horizontal transmission from larva to nymph or nymph to adult is essential for the release of Babesia into the host. The initiation of the asexual cycle of these parasites is different because Plasmodium sporozoites need to infect hepatocytes before egressed merozoites can infect erythrocytes, while Babesia sporozoites are known to enter the erythrocytic cycle directly. Plasmodium metabolism, as determined by its two- to threefold larger genome than different Babesia, is more complex. Plasmodium replication occurs in parasitophorous vacuole (PV) within the host cells, and a relatively large number of merozoites are released from each infected RBC after schizogony. The Babesia erythrocytic cycle lacks both PV and schizogony. Cytoadherence that allows the sequestration of Plasmodia, primarily P. falciparum in different organs facilitated by prominent adhesins, has not been documented for Babesia yet. Inflammatory immune responses contribute to the severity of malaria and babesiosis. Antibodies appear to play only a minor role in the resolution of these diseases; however, cellular and innate immunity are critical for the clearance of both pathogens. Inflammatory immune responses affect the severity of both diseases. Macrophages facilitate the resolution of both infections and also offer cross-protection against related protozoa. Although the immunosuppression of adaptive immune responses by these parasites does not seem to affect their own clearance, it significantly exacerbates diseases caused by coinfecting bacteria during coinfections.

Pathogens detected in the tick Haemaphysalis concinna in Western Poland: known and unknown threats

In recent years, a new focus of the relict tick Haemaphysalis concinna was discovered in Western Poland, near Wolsztyn, Greater Poland voivodeship. This species may play an important role in the circulation of pathogens of medical and veterinary importance. In the present study we tested 880 juvenile ticks collected from rodents, including 427 H. concinna, 443 Ixodes ricinus and 10 Dermacentor reticulatus for three of the most common pathogens vectored by ticks in Poland: Rickettsia and Babesia spp. and Borrelia burgdorferi s.l. Additionally, molecular techniques were applied for accurate identification of tick host species (the voles Microtus and Alexandromys). Our study found differences in the range and prevalence of vectored pathogens between the three tick species. DNA of all three pathogens was found in I. ricinus. In juvenile H. concinna, DNA of Babesia microti, Borrelia afzelii and Rickettsia sp. was identified. Moreover, DNA of a new unnamed Babesia species related to B. crassa, was found in two H. concinna nymphs. This genotype of Babesia was previously identified in H. concinna in the Far East and then in Central Europe. DNA of Rickettsia raoulti and B. afzelii was detected in D. reticulatus nymphs. Among rodent hosts, Alexandromys oeconomus seems to be host of the highest significance for juvenile tick stages and was the only host species with B. afzelii detected in blood samples. Using phylogenetic methods, we confirmed a clear division between rodents from the genera Microtus and Alexandromys. Moreover, we found that A. oeconomus trapped in Western Poland clustered with a Central European A. oeconomus allopatric phylogroup.

A Pilot Study on the Microbiome of Amblyomma hebraeum Tick Stages Infected and Non-Infected with Rickettsia africae.

Variation in tick microbiota may affect pathogen acquisition and transmission but for many vector species, including Amblyomma hebraeum, components and determinants of the microbiome are unidentified. This pilot study aimed to determine baseline microbial community within A. hebraeum nymphs infected- and non-infected with Rickettsia africae from the environment, and within adult ticks infected- and non-infected with R. africae collected from cattle sampled from two locations in the Eastern Cape province of South Africa. Adult A. hebraeum ticks (N = 13) and A. hebraeum nymph (N = 15) preliminary screened for R. africae were randomly selected and subjected to Illumina sequencing targeting the v3–v4 hypervariable regions of the 16S rRNA gene. No significant difference in microbial community composition, as well as rarefied OTU richness and diversity were detected between adults and nymphs. Nymphs showed a higher richness of bacterial taxa indicating blood-feeding could have resulted in loss of microbial diversity during the moulting stage from nymph to adult. Core OTUs that were in at least 50% of nymphs and adults negative and positive for Rickettsia at 1% minimum relative abundance were Rickettsia, Coxiella and Ruminococcaceae UCG-005 with a single genus Arsenophonus occurring only in nymphs negative for Rickettsia. Ehrlichia spp. was present in only four nymphal ticks positive for Rickettsia. Interestingly, Rickettsia aeschlimannii was found in one nymph and one adult, indicating the first ever detection of the species in A. hebraeum. Furthermore, A. hebraeum harboured a Coxiella-like endosymbiont, which should be investigated further as Coxiella may affect the viability and transmission of other organisms.

Distribution of Borrelia burgdorferi s.l. and Borrelia miyamotoi in Ixodes tick populations in Northern Germany, co-infections with Rickettsiales and assessment of potential influencing factors.

To determine Borrelia spp. (Spirochaetales: Spirochaetaceae) prevalence and species distribution in Northern Germany, Ixodes ticks were sampled from April to October in 2018 and 2019 by the flagging method at three locations each in five regions. Analysis by quantitative real-time PCR of 3150 individual ticks revealed an overall prevalence of 30.6%, without significant differences between tick stages (31.7% positive adults, 28.6% positive nymphs). Significant differences were observed in seasonal infection rates, but not between regions, landscape types or sampling years. Analysis of co-infections with Rickettsiales indicated a negative association between Borrelia and Anaplasma phagocytophilum infection. The most frequent Borrelia species differentiated by Reverse Line Blot were B. afzelii and B. garinii/B. bavariensis, followed by B. valaisiana, B. burgdorferi sensu stricto, B. spielmanii and B. lusitaniae. Furthermore, B. miyamotoi was identified in 12.9% of differentiable samples. No effect of region nor landscape type on species composition was found, but significant variations in the distribution at the different sampling sites within a region were observed. The detected monthly fluctuations in prevalence and the differences in intra-regional Borrelia species distribution underline the importance of long-term and multi-location monitoring of Borrelia spp. in ticks as an essential part of public health assessment.

Wild Small Mammals and Ticks in Zoos-Reservoir of Agents with Zoonotic Potential?

Wild small mammals and ticks play an important role in maintaining and spreading zoonoses in nature, as well as in captive animals. The aim of this study was to monitor selected agents with zoonotic potential in their reservoirs and vectors in a zoo, and to draw attention to the risk of possible contact with these pathogens. In total, 117 wild small mammals (rodents) and 166 ticks were collected in the area of Brno Zoo. Antibodies to the bacteria Coxiella burnetii, Francisella tularensis, and Borrelia burgdorferi s.l. were detected by a modified enzyme-linked immunosorbent assay in 19% (19/99), 4% (4/99), and 15% (15/99) of rodents, respectively. Antibodies to Leptospira spp. bacteria were detected by the microscopic agglutination test in 6% (4/63) of rodents. Coinfection (antibodies to more than two agents) were proved in 14.5% (15/97) of animals. The prevalence of C. burnetii statistically differed according to the years of trapping (p = 0.0241). The DNAs of B. burgdorferi s.l., Rickettsia sp., and Anaplasma phagocytophilum were detected by PCR in 16%, 6%, and 1% of ticks, respectively, without coinfection and without effect of life stage and sex of ticks on positivity. Sequencing showed homology with R. helvetica and A. phagocytophilum in four and one positive samples, respectively. The results of our study show that wild small mammals and ticks in a zoo could serve as reservoirs and vectors of infectious agents with zoonotic potential and thus present a risk of infection to zoo animals and also to keepers and visitors to a zoo.

Cytochrome b Drug Resistance Mutation Decreases Babesia Fitness in the Tick Stages But Not the Mammalian Erythrocytic Cycle.

Human babesiosis is an emerging tick-borne malaria-like illness caused by Babesia parasites following their development in erythrocytes. Here, we show that a mutation in the Babesia microti mitochondrial cytochrome b (Cytb) that confers resistance to the antibabesial drug ELQ-502 decreases parasite fitness in the arthropod vector. Interestingly, whereas the mutant allele does not affect B. microti fitness during the mammalian blood phase of the parasite life cycle and is genetically stable as parasite burden increases, ELQ-502-resistant mutant parasites developing in the tick vector are genetically unstable with a high rate of the wild-type allele emerging during the nymphal stage. Furthermore, we show that B. microti parasites with this mutation are transmitted from the tick to the host, raising the possibility that the frequency of Cytb resistance mutations may be decreased by passage through the tick vector, but could persist in the environment if present when ticks feed.

Lake Michigan insights from island studies: the roles of chipmunks and coyotes in maintaining Ixodes scapularis and Borrelia burgdorferi in the absence of white-tailed deer

Deer management (e.g., reduction) has been proposed as a tool to reduce the acarological risk of Lyme disease. There have been few opportunities to investigate Ixodes scapularis (blacklegged tick) and Borrelia burgdorferi sensu stricto dynamics in the absence of white-tailed deer (Odocoileus virginianus) in midwestern North America. A pair of islands in Lake Michigan presented a unique opportunity to study the role of alternative hosts for the adult stage of the blacklegged tick for maintaining a tick population as a deer herd exists on North Manitou Island but not on South Manitou Island, where coyotes (Canis latrans) and hares (Lepus americanus) are the dominant medium mammals. Additionally, we were able to investigate the maintenance of I. scapularis and B. burgdorferi in small mammal communities on both islands, which were dominated by eastern chipmunks (Tamias striatus). From 2011 to 2015, we surveyed both islands for blacklegged ticks by drag cloth sampling, bird mist netting, and small and medium-sized mammal trapping. We assayed questing ticks, on-host ticks, and mammal biopsies for the Lyme disease pathogen, B. burgdorferi. We detected all three life stages of the blacklegged tick on both islands. Of the medium mammals sampled, no snowshoe hares (Lepus americanus, 0/23) were parasitized by adult blacklegged ticks, but 2/2 coyotes (Canis latrans) sampled on South Manitou Island in 2014 were parasitized by adult blacklegged ticks, suggesting that coyotes played a role in maintaining the tick population in the absence of deer. We also detected I. scapularis ticks on passerine birds from both islands, providing support that birds contribute to maintaining as well as introducing blacklegged ticks and B. burgdorferi to the islands. We observed higher questing adult and nymphal tick densities, and higher B. burgdorferi infection prevalence in small mammals and in adult ticks on the island with deer as compared to the deer-free island. On the islands, we also found that 25% more chipmunks were tick-infested than mice, fed more larvae and nymphs relative to their proportional abundance compared to mice, and thus may play a larger role compared to mice in the maintenance of B. burgdorferi. Our investigation demonstrated that alternative hosts could maintain a local population of blacklegged ticks and an enzootic cycle of the Lyme disease bacterium in the absence of white-tailed deer. Thus, alternative adult blacklegged tick hosts should be considered when investigating deer-targeted management tools for reducing tick-borne disease risk, especially when the alternative host community may be abundant and diverse.