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

Background: 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. Methods: 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. Results: 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. Conclusions: 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.

A seven-legged tick - anomalous Ixodes ricinus female (Acari: Ixodidae) from Poland.

Morphological anomalies are considered a rare phenomenon among natural tick populations. New cases of abnormalities in ticks are being described, such as body assymetries, nanism, gynandromorphism and limb malformations. The tick removed from a cat was morphologically identified to species and developmental stage. The time of feeding on the host was determined. The specimen was tested using PCR and Real-Time PCR methods for the presence of the common tick-borne pathogens: Anaplasma phagocytophilum, Babesia spp, Borrelia spp., Neoehrlichia mikurensis, Rickettsia spp. For visualisation of the anomalous structures, scanning electron microscopy (SEM) was performed. The tick was identified as a slightly engorged adult female of I. ricinus exhibiting ectromely of leg I on the left side of the idiosoma. The specimen was tested positive for two medically important pathogens: A. phagocytophilum and N. mikurensis. The case report describes a rare case of a morphological anomaly in an I. ricinus tick from Poland.

Tick abundance and infection with three zoonotic bacteria are heterogeneous in a Belgian peri-urban forest.

Ixodes ricinus is a vector of several pathogens of public health interest. While forests are the primary habitat for I. ricinus, its abundance and infection prevalence are expected to vary within forest stands. This study assesses the spatio-temporal variations in tick abundance and infection prevalence with three pathogens in and around a peri-urban forest where human exposure is high. Ticks were sampled multiple times in 2016 and 2018 in multiple locations with a diversity of undergrowth, using the consecutive drags method. Three zoonotic pathogens were screened for, Borrelia burgdorferi s.l., Coxiella burnetii, and Francisella tularensis. The influence of season, type of site and micro-environmental factors on tick abundance were assessed with negative binomial generalized linear mixed-effects models. We collected 1642 nymphs and 181 adult ticks. Ticks were most abundant in the spring, in warmer temperatures, and where undergrowth was higher. Sites with vegetation unaffected by human presence had higher abundance of ticks. Forest undergrowth type and height were significant predictors of the level of tick abundance in a forest. The consecutive drags method is expected to provide more precise estimates of tick abundance, presumably through more varied contacts with foliage. Borrelia burgdorferi s.l. prevalence was estimated from pooled ticks at 5.33%, C. burnetii was detected in six pools and F. tularensis was not detected. Borrelia afzelii was the dominant B. burgdorferi genospecies. Tick abundance and B. burgdorferi s.l. infection prevalence were lower than other estimates in Belgian forests.

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.

Geographic variation in the distribution of Anaplasma phagocytophilum variants in host-seeking Ixodes scapularis nymphs and adults in the eastern United States elucidated using next generation sequencing.

Human anaplasmosis cases, caused by Anaplasma phagocytophilum, are increasing in the United States. This trend is explained, in part, by expansion in the geographic range of the primary vector, Ixodes scapularis. Multiple variants of A. phagocytophilum have been identified in field collected ticks, but only a single variant (human active, or "Ap-ha," variant) has been shown to be pathogenic in humans. Until recently, laboratory methods used to differentiate variants were cumbersome and seldomly used in large scale assessments of the pathogen's geographic distribution. As a result, many surveys reported A. phagocytophilum without segregating variants. Lack of discrimination among A. phagocytophilum variants could lead to overestimation of anaplasmosis risk to humans. Next Generation Sequencing (NGS) assays were recently developed to efficiently detect multiple Ixodes scapularis-borne human pathogens including Ap-ha. In this study, we utilized NGS to detect and differentiate A. phagocytophilum variants (Ap-ha vs. non ha) in host-seeking I. scapularis nymphs and adults collected across 23 states in the eastern United States from 2012 to 2023 as part of national tick surveillance efforts and research studies. Many of the included ticks were tested previously using a TaqMan PCR assay that could detect A. phagocytophilum but could not differentiate variants. We retested A. phagocytophilum infected ticks with NGS to differentiate variants. Anaplasma phagocytophilum (any variant) was identified in 165 (35 %) of 471 counties from which ticks were tested, whereas Ap-ha was detected in 70 (15 %) of 469 counties where variants were differentiated. Both variants were identified in 32 % (n = 40) of 126 counties with either variant detected. Among states where A. phagocytophilum (any variant) was detected, prevalence ranged from 2 % to 19 % in unfed adults and from 0.2 % to 7.8 % in unfed nymphs; prevalence of Ap-ha variant ranged from 0.0 % to 16 % in adults, and 0.0 % to 4.6 % in nymphs.

Phenology of five tick species in the central Great Plains.

The states of Kansas and Oklahoma, in the central Great Plains, lie at the western periphery of the geographic distributions of several tick species. As the focus of most research on ticks and tick-borne diseases has been on Lyme disease which commonly occurs in areas to the north and east, the ticks of this region have seen little research attention. Here, we report on the phenology and activity patterns shown by tick species observed at 10 sites across the two states and explore factors associated with abundance of all and life specific individuals of the dominant species. Ticks were collected in 2020-2022 using dragging, flagging and carbon-dioxide trapping techniques, designed to detect questing ticks. The dominant species was A. americanum (24098, 97%) followed by Dermacentor variabilis (370, 2%), D. albipictus (271, 1%), Ixodes scapularis (91, <1%) and A. maculatum (38, <1%). Amblyomma americanum, A. maculatum and D. variabilis were active in Spring and Summer, while D. albipictus and I. scapularis were active in Fall and Winter. Factors associated with numbers of individuals of A. americanum included day of year, habitat, and latitude. Similar associations were observed when abundance was examined by life-stage. Overall, the picture is one of broadly distributed tick species that shows seasonal limitations in the timing of their questing activity.

Statewide surveillance of Ixodes scapularis (Acari: Ixodidae) for the presence of the human pathogen Borrelia miyamotoi (Spirochaetales: Spirochaetaceae), a relapsing fever spirochete in Pennsylvania, USA, 2019-2020.

Borrelia miyamotoi disease is an emerging tick-borne human illness in the United States caused by Borrelia miyamotoi (Spirochaetales: Spirochaetaceae) bacterium. With Pennsylvania reporting thousands of tick-borne disease cases annually, determining the minimum infection rate (MIR) of B. miyamotoi in Ixodes scapularis (Say, Acari: Ixodidae) adults within Pennsylvania is of utmost importance. Active surveillance was performed from October 2019 to April 2020 to collect a minimum of 50 I. scapularis ticks from every county within Pennsylvania and then screened for B. miyamotoi via qPCR. Ticks were collected from all 67 counties with the majority of those being adult I. scapularis. Additional ticks collected were Dermacentor albipictus (Packard, Acari: Ixodidae), Haemaphysalis longicornis (Neumann, Acari: Ixodidae), and immature I. scapularis. Adult I. scapularis were pooled and tested for B. miyamotoi. MIR for positive B. miyamotoi pools and density of infected adult I. scapularis varied by county, with positive pools from 38 Pennsylvania counties. This is the first statewide evaluation of B. miyamotoi in Pennsylvania in questing adult I. scapularis. These prevalence and distribution data will aid health care practitioners within the state of Pennsylvania and the northeast United States to understand potential risk and bring awareness to the lesser known human Borrelia illness, Borrelia miyamotoi disease.

Refining Ixodes scapularis (Acari: Ixodidae) distribution models: a comparison of current methods to an established protocol.

Blacklegged ticks (Ixodes scapularis Say) pose an enormous public health risk in eastern North America as the vector responsible for transmitting 7 human pathogens, including those causing the most common vector-borne disease in the United States, Lyme disease. Species distribution modeling is an increasingly popular method for predicting the potential distribution and subsequent risk of blacklegged ticks, however, the development of such models thus far is highly variable and would benefit from the use of standardized protocols. To identify where standardized protocols would most benefit current distribution models, we completed the "Overview, Data, Model, Assessment, and Prediction" (ODMAP) distribution modeling protocol for 21 publications reporting 22 blacklegged tick distribution models. We calculated an average adherence of 73.4% (SD ± 29%). Most prominently, we found that authors could better justify and connect their selection of variables and associated spatial scales to blacklegged tick ecology. In addition, the authors could provide clearer descriptions of model development, including checks for multicollinearity, spatial autocorrelation, and plausibility. Finally, authors could improve their reporting of variable effects to avoid undermining the models' utility in informing species-environment relationships. To enhance future model rigor and reproducibility, we recommend utilizing several resources including the ODMAP protocol, and suggest that journals make protocol compliance a publication prerequisite.

Range of Ixodes laguri, a nidicolous tick that parasitizes critically endangered rodents, with details on its western distribution limit in Austria.

The nidicolous tick Ixodes laguri is a nest-dwelling parasite of small mammals that mainly infest rodents of the families Cricetidae, Gliridae, Muridae and Sciuridae. There is no proven vectorial role for I. laguri, although it is suggested that it is a vector of Francisella tularensis. In this study, a first map depicting the entire geographical distribution of I. laguri based on georeferenced locations is presented. For this purpose, a digital data set of 142 georeferenced locations from 16 countries was compiled. Particular attention is paid to the description of the westernmost record of I. laguri in the city of Vienna, Austria. There, I. laguri is specifically associated with its main hosts, the critically endangered European hamster (Cricetus cricetus) and the European ground squirrel (Spermophilus citellus). These two host species have also been mapped in the present paper to estimate the potential distribution of I. laguri in the Vienna metropolitan region. The range of I. laguri extends between 16-108∘ E and 38-54∘ N, i.e. from Vienna in the east of Austria to Ulaanbaatar, the capital of Mongolia. In contrast to tick species that are expanding their range and are also becoming more abundant as a result of global warming, I. laguri has become increasingly rare throughout its range. However, I. laguri is not threatened by climate change, but by anthropogenic influences on its hosts and their habitats, which are typically open grasslands and steppes. Rural habitats are threatened by the intensification of agriculture and semi-urban habitats are increasingly being destroyed by urban development.

Partial trailside Japanese barberry (Ranunculales: Berberidaceae) removal did not reduce the abundance of questing blacklegged ticks (Acari: Ixodidae).

In a nature reserve in southern Maine, we removed invasive Japanese barberry (Berberis thunbergii de Candolle) along sections of forested recreational trails that ran through dense barberry infestations. Barberry thickets provide questing substrate and a protective microclimate for blacklegged ticks (Ixodes scapularis Say), and trail users could brush up against encroaching barberry and acquire ticks. Trailside barberry removal will reduce or eliminate encroaching tick questing substrate and could reduce trailside questing tick abundance by creating a microclimate more hostile to ticks. The same-day cut-and-spray treatment comprised mechanical cutting of barberry clumps (individual plants with numerous ramets) followed immediately by targeted herbicide application to the resulting root crowns. The treatment created trail shoulders to a lateral width of 1-2 m on both sides of 100-m trail sections, with initial treatment in the fall of 2013 and one retreatment in the summer of 2014. Our aim was to remove 90% of barberry clumps to achieve a 50% or better reduction in questing tick abundance on trail shoulders. However, by the fall of 2015, there were only 41% fewer barberry clumps on treated vs. untreated trail sections and there was no reduction in either adults or nymphs. We concluded that our barberry treatment protocol was not sufficiently aggressive since the resulting ecotone habitat on trail shoulders proved suitable for questing I. scapularis. In principle, cutting back barberry along trails should reduce trail user contact with questing deer ticks, but we were unable to demonstrate a reduction in trailside tick abundance.

Cascading impacts of overstory structure in managed forests on understory structure, microclimate conditions, and Ixodes scapularis (Acari: Ixodidae) densities.

Forest management practices designed to meet varied landowner objectives affect wildlife habitat and may interrupt the life-cycle stages of disease vectors, including the black-legged tick, Ixodes scapularis Say (Acari: Ixodidae). Ixodes scapularis transmits multiple pathogens including Borrelia burgdorferi, the causative agent of Lyme disease, which is the most common tick-borne disease in the United States. There is evidence that a range of active forest management practices (e.g., invasive plant removal, prescribed burning) can alter tick densities and pathogen transmission. However, few studies have investigated relationships between forest stand structural variables commonly manipulated by timber harvesting and tick ecology. Foresters may harvest timber to create certain forest structural conditions like the mean number of trees, or basal area, per hectare. This study used a spatially replicated experiment in a blocked design to compare forest stands with a range of overstory structures and document variations in the midstory, understory, and forest floor, as well as microclimate conditions within tick off-host habitat. Greater numbers of trees or basal area per hectare correlated with greater canopy closure but less understory cover, stabilized microclimate temperature, higher microclimate humidity, and greater I. scapularis nymph densities. A random forest model identified understory forest structure as the strongest predictor of nymph densities. There was no relationship between the number of trees or basal area per hectare and daily deer (Odocoileus virginianus Zimmermann) activity or nymphal infection prevalence. These findings provide a deeper understanding of tick-habitat associations within a forest stand and have the potential to inform forest management decisions.

First record of Ixodes keiransi (Acari: Ixodidae) in New Jersey, USA.

The hard tick, Ixodes keiransi Beati, Nava, Venzal, & Guglielmone, formerly the North American lineage of Ixodes affinis Neumann, is expanding its range northward along the US East Coast. In July 2023, we collected I. keiransi adult female and nymph in a single sampling event, suggesting its range now includes southern New Jersey. In this area, I. keiransi is sympatric with northern populations of Ixodes scapularis Say (Acari: Ixodidae), the primary vector of Lyme disease. Given its status as an enzootic vector of spirochaetes in the Borrelia burgdorferi sensu lato complex, proper differentiation of these 2 species will be critical for accurate estimates of entomological risk. Targeted surveillance should be implemented to monitor further I. keiransi expansion and to elucidate the phenology and enzootic role of this and other understudied Ixodes spp. in the northeastern United States.

A Geographic Information System Approach to Map Tick Exposure Risk at a Scale for Public Health Intervention.

Tick-borne disease control and prevention have been largely ineffective compared to the control of other vector-borne diseases. Although control strategies exist, they are costly or ineffective at large spatial scales. We need tools to target these strategies to places of highest tick exposure risk. Here we present a geographic information system (GIS) method for mapping predicted tick exposure risk at a 200 m by 200 m resolution, appropriate for public health intervention. We followed the approach used to map tick habitat suitability over large areas. We used drag-cloth sampling to measure the density of nymphal blacklegged ticks (Ixodes scapularis, Say (Acari: Ixodidae)) at 24 sites in Addison and Rutland Counties, VT, United States. We used a GIS to average habitat, climatological, land-use/land-cover, and abiotic characteristics over 100 m, 400 m, 1,000 m, and 2,000 m buffers around each site to evaluate which characteristic at which buffer size best predicted density of nymphal ticks (DON). The relationships between predictor variables and DON were determined with random forest models. The 100 m buffer model performed best and explained 37.7% of the variation in DON, although was highly accurate at classifying sites as having below or above average DON. This model was applied to Addison County, VT, to predict tick exposure risk at a 200 m resolution. This GIS approach to map predicted DON over a small area with fine resolution, could be used to target public health campaigns and land management practices to reduce human exposure to ticks.

Beech tree masting explains the inter-annual variation in the fall and spring peaks of Ixodes ricinus ticks with different time lags.

BACKGROUND: The tick Ixodes ricinus is an important vector of tick-borne diseases including Lyme borreliosis. In continental Europe, the nymphal stage of I. ricinus often has a bimodal phenology with a large spring peak and a smaller fall peak. There is consensus about the origin of the spring nymphal peak, but there are two alternative hypotheses for the fall nymphal peak. In the direct development hypothesis, larvae quest as nymphs in the fall of the same year that they obtained their larval blood meal. In the developmental diapause hypothesis, larvae overwinter in the engorged state and quest as nymphs one year after they obtained their larval blood meal. These two hypotheses make different predictions about the time lags that separate the larval blood meal and the density of questing nymphs (DON) in the spring and fall. METHODS: Inter-annual variation in seed production (masting) by deciduous trees is a time-lagged index for the density of vertebrate hosts (e.g., rodents) which provide blood meals for larval ticks. We used a long-term data set on the masting of the European beech tree and a 15-year study on the DON at 4 different elevation sites in western Switzerland to differentiate between the two alternative hypotheses for the origin of the fall nymphal peak. RESULTS: Questing I. ricinus nymphs had a bimodal phenology at the three lower elevation sites, but a unimodal phenology at the top elevation site. At the lower elevation sites, the DON in the fall was strongly correlated with the DON in the spring of the following year. The inter-annual variation in the densities of I. ricinus nymphs in the fall and spring was best explained by a 1-year versus a 2-year time lag with the beech tree masting index. Fall nymphs had higher fat content than spring nymphs indicating that they were younger. All these observations are consistent with the direct development hypothesis for the fall peak of I. ricinus nymphs at our study site. Our study provides new insight into the complex bimodal phenology of this important disease vector. CONCLUSIONS: Public health officials in Europe should be aware that following a strong mast year, the DON will increase 1 year later in the fall and 2 years later in the spring. Studies of I. ricinus populations with a bimodal phenology should consider that the spring and fall peak in the same calendar year represent different generations of ticks.

Duration of tick attachment necessary for transmission of Anaplasma phagocytophilum by Ixodes scapularis (Acari: Ixodidae) nymphs.

This study assessed the duration of tick attachment necessary for a successful transmission of Anaplasma phagocytophilum by an infected I. scapularis nymph. Individual nymphs were placed upon BALB/c mice and allowed to feed for predetermined time intervals of 4 to 72 h. Ticks removed from mice at predetermined intervals were tested by PCR for verification of infection and evaluation of the bacterial load. The success of pathogen transmission to mice was assessed by blood-PCR at 7, 14 and 21 days postinfestation, and IFA at 21 days postinfestation. Anaplasma phagocytophilum infection was documented in 10-30 % of mice, from which ticks were removed within the first 20 h of feeding. However, transmission success was ≥70% if ticks remained attached for 36 h or longer. Notably, none of the PCR-positive mice that were exposed to infected ticks for 4 to 8 h and only half of PCR-positive mice exposed for 24 h developed antibodies within 3 weeks postinfestation. On the other hand, all mice with detectable bacteremia after being infested for 36 h seroconverted. This suggests that although some of the ticks removed prior to 24 h of attachment succeed in injecting a small amount of A. phagocytophilum, this amount is insufficient for stimulating humoral immunity and perhaps for establishing disseminated infection in BALB/c mice. Although A. phagocytophilum may be present in salivary glands of unfed I. scapularis nymphs, the amount of A. phagocytophilum initially contained in saliva appears insufficient to cause sustainable infection in a host. Replication and, maybe, reactivation of the agent for 12-24 h in a feeding tick is required before a mouse can be consistently infected.

Absence of Francisella tularensis in Finnish Ixodes ricinus and Ixodes persulcatus ticks.

Francisella tularensis subsp. holarctica is the causative agent of tularaemia in Europe. Finland is a high-incidence region for tularaemia, with mosquito bites as the most common sources of infection. However, in Central and Western Europe, ticks (Acari: Ixodidae) have been suggested as the main vectors. Indeed, several studies have reported the pathogen from the locally most common human-biting tick species, Ixodes ricinus. In Finland, the occurrence of the pathogen in ticks has started receiving attention only recently. Here, we collate previous tick screening data from Finland regarding F. tularensis as well as present the results from a novel screening of roughly 15 000 I. ricinus and I. persulcatus collected from across the country. In total, 14 878 ticks collected between 2015 and 2020 were screened for F. tularensis using a TaqMan-based qPCR assay targeting the 23 KDa gene. The combined screening efforts of the current and previous studies, encompassing roughly 20 000 ticks, did not find any positive ticks. Given the negative results despite the considerable sample size, it appears that the pathogen is not circulating in local tick populations in Finland. We discuss some possible reasons for the lack of the bacterium in ticks in this high-incidence region of tularaemia.

Integrated Tick Management in South Central Wisconsin: Impact of Invasive Vegetation Removal and Host-Targeted Acaricides on the Density of Questing Ixodes scapularis (Acari: Ixodidae) Nymphs.

As tick-borne disease incidence increases and pathogens expand into new areas, the need for effective tick management strategies is paramount. In this 5-yr study (2014-2018) conducted in south central Wisconsin, we assessed whether an integrated tick management approach, deployed during peak tick activity (May-August), was more effective at reducing black-legged ticks (Ixodes scapularis Say (Ixodida: Ixodidae)), than individual interventions. Using a factorial design, invasive vegetation removal (Amur honeysuckle, Lonicera maackii Ruprecht (Dipsacales: Caprifoliaceae) and common buckthorn, Rhamnus cathartica Linnaeus (Rosales: Rhamnaceae)) was coupled with deployments of permethrin-treated cotton nesting materials (tick tubes) that target the white-footed mouse (Peromyscus leucopus Rafinesque (Rodentia: Cricetidae)). Results show that the probability of encountering a larval tick by drag sampling was unaffected by treatments at the cumulative 5-yr level. However, vegetation removal significantly reduced larval encounters in 2014, 2015, and 2018, by 33%, 57%, and 61% respectively, and reduced the density of questing nymphal (DON) ticks by 45% in 2015 compared to controls. Despite the limited effect on DON, vegetation removal significantly reduced the cumulative 5-yr density of Borrelia burgdorferi sensu stricto infected nymphs (DIN) (70%) compared to controls as a result of decreased nymphal infection prevalence. Sites treated with tick tubes had lower DIN (66%) and DON (54%) across the study and nymphs were reduced every year following the initial year of deployment compared to controls. Combining treatments did not further reduce DIN or DONs. We conclude that long-term integration of tick tubes with invasive vegetation removal does not provide additional benefit over individual treatments alone.

Free-living hard ticks (Ixodida: Ixodidae) from three different natural environments of Costa Rica.

This paper presents data on free-living ticks collected by flagging and using CO2 traps in three natural areas in Costa Rica: Carara National Park (CNP), Palo Verde National Park (PVNP), and a Private Forest Reserve in Sarapiquí (SPR). Data were analyzed calculating aspects of alpha diversity (species richness, entropy; dominance index, and evenness); and for beta diversity, compositional similarity between communities of ticks was also calculated. We collected 12,795 ticks belonging to 10 species: Amblyomma coelebs, Amblyomma dissimile, Amblyomma mixtum, Amblyomma naponense, Amblyomma cf. oblongoguttatum, Amblyomma cf. parvum, Amblyomma sabanerae, Amblyomma tapirellum, Haemaphysalis juxtakochi and Ixodes affinis. The number of species and individuals varied between sites: 5970 ticks were collected in CNP, 4443 in PVNP, and 2382 in SPR. Amblyomma cf. oblongoguttatum and A. cf. parvum were collected at all three sites, but A. mixtum was the most abundant species, even though it was not collected in SPR. Values of alpha diversity were calculated for CNP and SPR, while diversity in PVNP was the lowest of the three locations. Evenness was highest in SPR and lowest in CNP. The only community that presented dominance was PVNP. Beta diversity showed low similarity between the three locations with the lowest being CNP and SPR. For the three localities, estimates of the number of tick species based on presence/absence data was higher using flagging than CO2; and considering the stage of the ticks collected. More larvae were captured using CO2 traps than by flagging, while flagging was better for collecting adults. To our knowledge this is the first study in Costa Rica that compares these two sampling methods in three different environmental areas.

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.

Double anus in an Ixodes scapularis nymph, a medically important tick vector.

BACKGROUND: Ixodes scapularis ticks are medically important arthropod vectors that transmit several pathogens to humans. The observations of morphological abnormalities, including nanism, missing leg, extra leg, and gynandromorphism, have been reported in these ticks. In this study, we report the presence of two anuses in a laboratory-reared I. scapularis nymph. RESULTS: Larval ticks were allowed to feed on mice and to molt to nymphs. Two anuses were observed in one of the freshly molted nymphs. Stereo and scanning electron microscopy confirmed the presence of two anuses in one nymph within a single anal groove. CONCLUSIONS: This report confirms the rare occurrence of double anus in I. scapularis.