Epidemiology of ticks submitted from human hosts in Alberta, Canada (2000-2019).

ABSTRACTThe geographic range and occurrence of tick species is dynamic. This has important public health implications due to important tick species that can transmit pathogens. This study presents a retrospective review of tick genera recovered from humans and submitted for identification in Alberta, Canada, over a 19-year period. The total number of ticks and proportion of genera were analyzed over time. Molecular testing for a number of pathogens associated with Ixodes scapularis and I. pacificus was conducted. A total of 2,358 ticks were submitted between 2000 and 2019, with 98.6% being acquired in Alberta. The number of ticks submitted increased significantly over time (p < 0.0001). Dermacentor ticks were the most abundant genus, followed by Ixodes and Amblyomma. There was a significant decrease in the proportion of Dermacentor ticks between 2013 and 2019 (p = 0.02), with a corresponding increase in the proportion of Ixodes ticks over the same time (p = 0.04). No statistically significant change in seasonality was identified. Borrelia burgdorferi was detected in 8/76 (10.5%; 95% CI 5.4-19.4%) of all I. scapularis and I. pacificus ticks submitted. This translated to a B. burgdorferi positivity of 0.35% (95% CI 0.15-0.68%) among all ticks received. Dermacentor species (especially D. andersoni) remains the most common tick feeding on humans in Alberta. Small numbers of vector species (including I. scapularis/pacificus) are encountered annually over widely separated geographic areas in the province. The risk of exposure to tick-borne pathogens (e.g. Lyme disease) in Alberta remains low.

Identification of Hard Ticks in the United States: A Practical Guide for Clinicians and Pathologists.

ABSTRACT: According to guidelines published by the Infectious Disease Society of America, Lyme disease prophylaxis is possible if a tick can be identified as Ixodes scapularis (nymphal or adult) within 72 hours of tick removal. However, a recent survey of medical practitioners indicates generally poor proficiency in tick identification. In this study, we provide a simple, practical guide to aid medical practitioners in identifying the most commonly encountered human biting ticks of North America.

Proficiency at Tick Identification by Pathologists and Clinicians Is Poor.

OBJECTIVES: Prompt accurate identification of tick species is required for appropriate administration of single dose antimicrobial prophylaxis for Lyme disease in selected patients. To determine the proficiency of clinicians at tick identification in the northeastern United States where Lyme disease has its highest incidence, we undertook a survey. METHODS: We analyzed the results of a voluntary survey testing proficiency in identifying tick species using high-resolution photographs of ticks. RESULTS: Only 35% of ticks were correctly identified. Although 60% of respondents could identify a nonengorged adult blacklegged tick, only 34% could correctly identify a partially engorged blacklegged tick. Participants performed even worse at classifying brown dog, American dog, and Lone Star ticks. CONCLUSIONS: Proficiency of tick identification by pathologists and clinicians is poor.

Occurrence of tick-borne pathogens in questing Ixodes ricinus ticks from Wester Ross, Northwest Scotland.

BACKGROUND: Lyme borreliosis and other tick-borne diseases emerge from increased interactions between humans, other animals, and infected ticks. The risk of acquiring a tick-borne infection varies across space and time, so knowledge of the occurrence and prevalence of pathogens in ticks can facilitate disease diagnosis in a specific area and the implementation of mitigation measures and awareness campaigns. Here we identify the occurrence and prevalence of several pathogens in Ixodes ricinus ticks in Wester Ross, Northwest Scotland, a region of high tourism and tick exposure, yet data-poor in terms of tick-borne pathogens. METHODS: Questing I. ricinus nymphs (n = 2828) were collected from 26 sites in 2018 and 2019 and tested for the presence of tick-borne pathogens using PCR-based methods. Prevalence was compared with other regions of Scotland, England, Wales, and the Netherlands. RESULTS: Anaplasma phagocytophilum (4.7% prevalence), Borrelia burgdorferi sensu lato (s.l.) (2.2%), Babesia from clade X (0.2%), Rickettsia helvetica (0.04%), and Spiroplasma ixodetis (0.4%) were detected, but no Neoehrlichia mikurensis, Borrelia miyamotoi, or Babesia microti. Typing of A. phagocytophilum using a fragment of the GroEL gene identified the presence of both ecotype I and ecotype II. Genospecies identification of Borrelia burgdorferi s.l. revealed B. afzelii (53% of infected nymphs), B. garinii (9%), B. burgdorferi sensu stricto (7%), and B. valaisiana (31%). We found similar prevalence of A. phagocytophilum in Wester Ross as in the Netherlands, but higher than in other parts of Great Britain. We found lower B. burgdorferi s.l. prevalence than in England or the Netherlands, and similar to some other Scottish studies. We found higher prevalence of B. valaisiana and lower prevalence of B. garinii than in other Scottish studies. We found S. ixodetis at much lower prevalence than in the Netherlands, and R. helvetica at much lower prevalence than in England and the Netherlands. CONCLUSIONS: As far as we know, this is the first description of S. ixodetis in Great Britain. The results are relevant for disease surveillance and management for public and veterinary health. The findings can also aid in designing targeted public health campaigns and in raising awareness among outdoor recreationists and professionals.

Vectra 3D (dinotefuran, pyriproxyfen and permethrin) prevents acquisition of Borrelia burgdorferi sensu stricto by Ixodes ricinus and Ixodes scapularis ticks in an ex vivo feeding model.

BACKGROUND: We evaluated the efficiency of an ex vivo feeding technique using a silicone membrane-based feeding chamber to (i) assess the anti-feeding and acaricidal efficacy of a spot-on combination of dinotefuran, pyriproxyfen and permethrin (DPP, Vectra® 3D) against adult Ixodes scapularis and Ixodes ricinus ticks, and to (ii) explore its effect on blocking the acquisition of Borrelia burgdorferi sensu stricto. METHODS: Eight purpose-bred dogs were randomly allocated to two equal-size groups based on body weight assessed on day 2. DPP was administered topically, as spot-on, to four dogs on day 0. Hair from the eight dogs was collected individually by brushing the whole body on days 2, 7, 14, 21, 28 and 35. On each day of hair collection, 0.05 g of sampled hair was applied on the membrane corresponding to each feeding unit (FU). Seventy-two FU were each seeded with 30 adults of I. scapularis (n = 24 FU) or I. ricinus ticks (n = 48 FU). Bovine blood spiked with B. burgdorferi sensu stricto (strain B31) was added into each unit and changed every 12 h for 4 days. Tick mortality was assessed 1 h after seeding. One additional hour of incubation was added for live/moribund specimens and reassessed for viability. All remaining live/moribund ticks were left in the feeders and tick engorgement status was recorded at 96 h after seeding, and the uptake of B. burgdorferi s.s. was examined in the collected ticks by applying quantitative real-time PCR. RESULTS: Exposure to DPP-treated hair was 100% effective in blocking B. burgdorferi s.s. acquisition. The anti-feeding efficacy remained stable (100%) against both Ixodes species throughout the study. The acaricidal efficacy of DPP evaluated at 1 and 2 h after exposure was 100% throughout the study for I. ricinus, except the 1-h assessment on day 28 (95.9%) and day 35 (95.3%). The 1-h assessment of acaricidal efficacy was 100% at all time points for I. scapularis. CONCLUSIONS: The ex vivo feeding system developed here demonstrated a protective effect of DPP against the acquisition of B. burgdorferi without exposing the animals to the vectors or to the pathogen.

Identification of tick-borne pathogens by metagenomic next-generation sequencing in Dermacentor nuttalli and Ixodes persulcatus in Inner Mongolia, China.

BACKGROUND: Hard ticks act as arthropod vectors in the transmission of human and animal pathogens and are widely distributed in northern China. The aim of this study is to screen the important tick-borne pathogens (TBPs) carried by hard ticks in Inner Mongolia using metagenomic next-generation sequencing (mNGS) and to estimate the risk of human infection imposed by tick bites. METHODS: The adult Dermacentor nuttalli (n = 203) and Ixodes persulcatus (n = 36) ticks feeding on cattle were collected. The pooled DNA samples prepared from these ticks were sequenced as the templates for mNGS to survey the presence of TBPs at the genus level. Individual tick DNA samples were detected by genus--specific or group-specific nested polymerase chain reaction (PCR) of these TBPs and combined with DNA sequencing assay to confirm the results of mNGS. RESULTS: R. raoultii (45.32%, 92/203), Candidatus R. tarasevichiae (5.42%, 11/203), Anaplasma sp. Mongolia (26.60%, 54/203), Coxiella-like endosymbiont (CLE) (53.69%, 109/203), and Babesia venatorum (7.88%, 16/203) were detected in D. nuttalli, while R. raoultii (30.56%, 11/36), Anaplasma sp. Mongolia (27.80%, 10/36), and CLE (27.80%, 10/36) were detected in I. persulcatus. The double- and triple-pathogen/endosymbiont co-infections were detected in 40.39% of D. nuttalli and 13.89% of I. persulcatus, respectively. The dual co-infection with R. raoultii and CLE (14.29%, 29/203) and triple co-infection with R. raoultii, Anaplasma sp. Mongolia, and CLE (13.79%, 28/203) were most frequent in D. nuttalli. CONCLUSIONS: This study provides insight into the microbial diversity of D. nuttalli and I. persulcatus in Inner Mongolia, China, reporting for the first time that Candidatus R. tarasevichiae had been found in D. nuttalli in China, and for the first time in the world that Anaplasma sp. Mongolia has been detected in I. persulcatus. This study proves that various vertically transmitted pathogens co-inhabit D. nuttalli and I. persulcatus, and indicates that cattle in Inner Mongolia are exposed to several TBPs.

Focus Stacking Images of Morphological Character States for Differentiating the Adults of Ixodes affinis and Ixodes scapularis (Acari: Ixodidae) in Areas of Sympatry.

Adult females and males of Ixodes affinis and Ixodes scapularis are illustrated by focus stacking image photography, and morphological character states are described that reliably differentiate the two species. In conjunction with other environmental cues, such as the questing phenology of adults, these characteristics will enable the rapid identification of adults of either sex along the southern Coastal Plain of the United States, where these species are sympatric.

Ticks infesting humans and associated pathogens: a cross-sectional study in a 3-year period (2017-2019) in northwest Italy.

BACKGROUND: Tick-borne diseases are common throughout Europe. Ticks transmit pathogens to the host while feeding and together with mosquitoes, they are major vectors of infectious agents worldwide. In recent years, there has been a marked increase in the incidence of tick-bite events and tick-borne disease in northwest Italy, but information on the prevalence of tick-borne pathogens in ticks removed from humans remains scarce. To fill this gap, we report here the prevalence of tick bites and tick-borne pathogens documented for humans in Piedmont, northwest Italy, in the 3-year period 2017-2019. METHODS: Ticks attached to humans during 2017-2019 were collected from residents of urban and rural area by physicians and veterinarians working with local veterinary agencies. All ticks (n = 1290) were morphologically identified to the species level. A subset of ticks removed from children (age 0-18 years) and the elderly (> 70 years), both age groups considered to be at-risk populations, was screened by biomolecular analysis to detect pathogens (e.g. Rickettsia spp., Borrelia spp., Anaplasma spp.). Pathogen identity was confirmed by Sanger sequencing. RESULTS: Ticks were taxonomically assigned to ten species of six genera (Amblyomma, Dermacentor, Haemaphysalis, Hyalomma, Ixodes and Rhipicephalus). Most belonged to the genus Ixodes: 1009 ticks (78.22%) were classified as Ixodes ricinus. A subset of 500 ticks collected from the two at-risk populations were subjected to PCR assay to determine the presence of Rickettsia spp., Borrelia spp., and Anaplasma spp. The overall prevalence of infection was 22.8% (n = 114; 95% confidence interval [CI]: 19.19-26.73%), meaning that at least one pathogen was detected: Rickettsia spp. (prevalence 15%, n = 76; 95% CI 12.17-18.65%); Borrelia spp. (prevalence 6.4%, n = 32; 95% CI 4.42-8.92%); and Anaplasma spp. (prevalence 1.2%, n = 6; 95% CI 0.44-2.6%). CONCLUSIONS: Our data underline the importance of surveillance in the epidemiology of tick-borne diseases and the implementation of strategies to control tick infestation and associated pathogens.

High-elevational occurrence of two tick species, Ixodes ricinus and I. trianguliceps, at their northern distribution range.

BACKGROUND: During the last decades a northward and upward range shift has been observed among many organisms across different taxa. In the northern hemisphere, ticks have been observed to have increased their latitudinal and altitudinal range limit. However, the elevational expansion at its northern distribution range remains largely unstudied. In this study we investigated the altitudinal distribution of the exophilic Ixodes ricinus and endophilic I. trianguliceps on two mountain slopes in Norway by assessing larval infestation rates on bank voles (Myodes glareolus). METHODS: During 2017 and 2018, 1325 bank voles were captured during the spring, summer and autumn at ten trapping stations ranging from 100 m to 1000 m.a.s.l. in two study areas in southern Norway. We used generalized logistic regression models to estimate the prevalence of infestation of both tick species along gradients of altitude, considering study area, collection year and season, temperature, humidity and altitude interactions as extrinsic variables, and host body mass and sex as intrinsic predictor variables. RESULTS: We found that both I. ricinus and I. trianguliceps infested bank voles at altitudes up to 1000 m.a.s.l., which is a substantial increase in altitude compared to previous findings for I. ricinus in this region. The infestation rates declined more rapidly with increasing altitude for I. ricinus compared to I. trianguliceps, indicating that the endophilic ecology of I. trianguliceps may provide shelter from limiting factors tied to altitude. Seasonal effects limited the occurrence of I. ricinus during autumn, but I. trianguliceps was found to infest rodents at all altitudes during all seasons of both years. CONCLUSIONS: This study provides new insights into the altitudinal distribution of two tick species at their northern distribution range, one with the potential to transmit zoonotic pathogens to both humans and livestock. With warming temperatures predicted to increase, and especially so in the northern regions, the risk of tick-borne infections is likely to become a concern at increasingly higher altitudes in the future.

Benefits and Drawbacks of Citizen Science to Complement Traditional Data Gathering Approaches for Medically Important Hard Ticks (Acari: Ixodidae) in the United States.

Tick-borne diseases are increasing in North America. Knowledge of which tick species and associated human pathogens are present locally can inform the public and medical community about the acarological risk for tick bites and tick-borne infections. Citizen science (also called community-based monitoring, volunteer monitoring, or participatory science) is emerging as a potential approach to complement traditional tick record data gathering where all aspects of the work is done by researchers or public health professionals. One key question is how citizen science can best be used to generate high-quality data to fill knowledge gaps that are difficult to address using traditional data gathering approaches. Citizen science is particularly useful to generate information on human-tick encounters and may also contribute to geographical tick records to help define species distributions across large areas. Previous citizen science projects have utilized three distinct tick record data gathering methods including submission of: 1) physical tick specimens for identification by professional entomologists, 2) digital images of ticks for identification by professional entomologists, and 3) data where the tick species and life stage were identified by the citizen scientist. We explore the benefits and drawbacks of citizen science, relative to the traditional scientific approach, to generate data on tick records, with special emphasis on data quality for species identification and tick encounter locations. We recognize the value of citizen science to tick research but caution that the generated information must be interpreted cautiously with data quality limitations firmly in mind to avoid misleading conclusions.

Ixodes silvanus n. sp. (Acari: Ixodidae), a new member of the subgenus Trichotoixodes Reznik, 1961 from northwestern Argentina.

Females, nymphs, and larvae of Ixodes silvanus n. sp. collected from birds and from the vegetation in northwestern Argentina (Yungas Phytogeographic Province) are described herein. The new species belongs to the subgenus Trichotoixodes (Acari: Ixodidae). The female is diagnosed by a combination of the following characters: scutum with setae moderately long and more numerous in central field, fewer and moderately long setae on lateral fields, and inconspicuous setae in anterior field; basis capituli subtriangular dorsally; porose areas large and irregular in shape, lacking distinct margins; auriculae with straight edges diverging posterolaterally and ending with small blunt processes; hypostome narrow and pointed with dental formula 4/4 in the anterior third, then 3/3 and 2/2 near the base; coxae I with two spurs, sub-equal in size, internal slightly slimmer than external. The nymph is diagnosed by notum with numerous and long setae, ventral surface covered by numerous whitish setae, scutum with short scapulae and few and shallow punctations, setae on scutum few, short and irregularly distributed, basis capituli sub-triangular dorsally with posterior margin straight, cornua large and directed postero-laterally, auriculae large and projected laterally, lateral margin of basis capituli above auriculae with a lateral and triangular projection, hypostome pointed with dental formula 3/3 in the anterior third and then 2/2, and coxa I with two short, sub-equal, triangular spurs. The diagnostic characters of the larva are: basis capituli dorsally sub-triangular with lateral angles acute and posterior margin straight, auriculae as large triangular lateral projections, hypostome with apex bluntly pointed and dental formula 3/3 in the anterior third and then 2/2, coxa I with two short, sub-equal, triangular spurs, and pattern of dorsal and ventral body setae. This new species is phylogenetically related to Ixodes brunneus, Ixodes turdus and Ixodes frontalis, and the principal hosts for all its parasitic stages are birds.

Ixodes spp. from Dogs and Cats in the United States: Diversity, Seasonality, and Prevalence of Borrelia burgdorferi and Anaplasma phagocytophilum.

Ixodes spp. are commonly found on dogs and cats throughout the world. In the eastern United States, 16S rDNA sequence of Ixodes scapularis, the predominant species, reveals two clades-American and Southern. To confirm the species and clades of Ixodes spp. ticks submitted from pets, we examined ticks morphologically and evaluated 16S rDNA sequence from 500 ticks submitted from 253 dogs, 99 cats, 1 rabbit, and 1 ferret from 41 states. To estimate pathogen prevalence, flaB of Borrelia burgdorferi (Bb) sensu stricto and 16S rDNA of Anaplasma phagocytophilum (Ap) were amplified and sequenced. Most Ixodes spp. from the Northeast (n = 115/115; 100%) and the Midwest (n = 77/80; 96.3%) were I. scapularis, American clade. Borrelia spp. were identified in 34 of 192 (17.8%) and Ap in 5 of 192 (2.6%) I. scapularis. Two Ixodes cookei and one Ixodes texanus were identified from Ohio, Illinois, and Michigan. In contrast, 156 of 261 (59.8%) Ixodes spp. from the Southeast were I. scapularis, American clade; 86 of 261 (33.0%) were I. scapularis, Southern clade; 9 of 261 (3.4%) were Ixodes affinis; and 10 of 261 (3.8%) were I. cookei. Southern clade was significantly more common in Florida and less common in the upper South (p < 0.0001). One I. scapularis (1/242; 0.4%) from the Southeast (Kentucky) tested positive for Bb and 6 of 242 (2.5%) were positive for Ap. In the West, most (34/44; 77.3%) Ixodes spp. were Ixodes pacificus, with Ixodes angustus (n = 6) submitted from dogs in Alaska, Washington, and Oregon and Ixodes haerlei (n = 4) preliminarily identified from a dog in Montana. Pathogens were not detected in any ticks from the West. Although I. scapularis, American clade, predominated in the Northeast and Midwest, additional Ixodes spp. were found on dogs and cats in other regions and pathogens were less commonly detected. The role of less common Ixodes spp. as disease vectors, if any, warrants continued investigation.

Migratory birds as disseminators of ticks and the tick-borne pathogens Borrelia bacteria and tick-borne encephalitis (TBE) virus: a seasonal study at Ottenby Bird Observatory in South-eastern Sweden.

BACKGROUND: Birds can act as reservoirs of tick-borne pathogens and can also disperse pathogen-containing ticks to both nearby and remote localities. The aims of this study were to estimate tick infestation patterns on migratory birds and the prevalence of different Borrelia species and tick-borne encephalitis virus (TBEV) in ticks removed from birds in south-eastern Sweden. METHODS: Ticks were collected from resident and migratory birds captured at the Ottenby Bird Observatory, Öland, Sweden, from March to November 2009. Ticks were molecularly identified to species, and morphologically to developmental stage, and the presence of Borrelia bacteria and TBEV was determined by quantitative real-time PCR. RESULTS: A total of 1339 ticks in the genera Haemaphysalis, Hyalomma, and Ixodes was recorded of which I. ricinus was the most abundant species. Important tick hosts were the European robin (Erithacus rubecula), Blackbird (Turdus merula), Tree pipit (Anthus trivialis), Eurasian wren (Troglodytes troglodytes), Common redstart (Phoenicurus phoenicurus), Willow warbler (Phylloscopus trochilus), and Common whitethroat (Sylvia communis). Borrelia bacteria were detected in 25% (285/1,124) of the detached ticks available for analysis. Seven Borrelia species (B. afzelii, B. burgdorferi (s.s.), B. garinii, B. lusitaniae, B. turdi, B. valaisiana, and B. miyamotoi) were identified. B. turdi was recorded for the first time in ticks in Sweden. The number of Borrelia cells per tick ranged from 2.0 × 100 to 7.0 × 105. B. miyamotoi-containing ticks contained a significantly higher median number of Borrelia cells than B. burgdorferi (s.l.)-containing ticks. B. garinii and B. miyamotoi were the most prevalent Borrelia species in tick larvae. Larvae of I. ricinus with B. garinii were removed from seven bird species, particularly S. communis and A. trivialis, which may suggest that the larvae had contracted the Borrelia bacteria from or via these birds. Also, a high percentage of tick larvae containing B. miyamotoi was removed from E. rubecula. All ticks were negative for TBEV. CONCLUSIONS: The results corroborate the view that the contributions of birds to human disease are substantial, particularly as blood hosts for ticks and for their short-, medium-, and long-distance dispersal. Moreover, several ground-foraging bird species appear to be important for the maintenance and dispersal of Borrelia species. The absence of TBEV in the ticks conforms to other similar studies.

Description of three new species of Ixodes Latreille, 1795 (Acari: Ixodidae), parasites of tenrecs (Afrotheria: Tenrecidae) on Madagascar.

Ixodes soarimalalae n. sp., Ixodes uilenbergi n. sp. and Ixodes uncus n. sp. (Acari: Ixodidae), are described based on females ex various species of tenrecs (Afrosoricida: Tenrecidae) from Madagascar. Females of all of these new species are similar to those of other species of the subgenus Afrixodes Morel, 1966 known from Madagascar, from which they can be distinguished and from one other by the size of scutum, size of scutal setae, shape of alloscutal setae, shape of genital aperture, development of genital apron, size of auriculae, size of anterior angle of basis capituli, size of palpi, dental formula on hypostome and size and development of spurs on coxa I.

Description of five new species of Ixodes Latreille, 1795 (Acari: Ixodidae) and redescription of I. luxuriosus Schulze, 1935, I. steini Schulze, 1935 and I. zaglossi Kohls, 1960, parasites of marsupials, rodents and echidnas in New Guinea Island.

Ixodes acer n. sp., Ixodes giluwensis n. sp., Ixodes mirzai n. sp., Ixodes planiscutatus n. sp. and Ixodes stellae n. sp. (Acari: Ixodidae), are described based on females ex various marsupials (Dasyuromorphia: Dasyuridae, Diprotodontia: Petauridae, Peramelemorphia: Peramelidae), rodents (Rodentia: Muridae) and a bird (Galliformes: Megapodiidae) from Indonesia (Papua Province) and Papua New Guinea. Females of all new species are similar to those of I. luxuriosus Schulze, 1935, I. steini Schulze, 1935 and I. zaglossi Kohls, 1960, but can be distinguished from them and each other by the distinctness of lateral carinae and cervical grooves on the scutum, length of setae on the idiosoma, sculpture of the basis capituli and palpal segment I dorsally, size of the dorsal cornua, shape and sculpture of the palpal segment I ventrally, size of ventral spur on the palpal segment I, dental formula on the hypostome and measurements of various structures and their proportions. For comparative purposes the female of I. luxuriosus, I. steini and I. zaglossi is redescribed and lectotypes for I. luxuriosus and I. steini have been designated. Studied females of I. luxuriosus, I. steini and I. zaglossi were found on marsupials (Dasyuromorphia: Dasyuridae; Diprotodontia: Macropodidae, Petauridae, Phalangeridae, Pseudocheiridae; Peramelemorphia: Peramelidae), rodents (Rodentia: Muridae) and echidnas (Monotremata: Tachyglossidae) in Indonesia (Papua Province) and Papua New Guinea.

Invasive rabbits host immature Ixodes ticks at the urban-forest interface.

Introduced wildlife may be important alternative hosts for generalist ticks that cause health issues for humans and companion animals in urban areas, but to date are rarely considered as part of the tick-host community compared to native wildlife. In Australia, European rabbits, Oryctolagus cuniculus, are a widespread and abundant invasive species common to a range of human-modified ecosystems. To understand the potential role of rabbits in the life cycle of Australian ticks, we investigated the seasonal abundance of all tick life stages (larva, nymph, and adult) on rabbits collected from pest control programs in two urban forest remnants in Sydney, Australia. We also recorded whether larvae, nymphs, and adults were attached to the head, body, or limbs of rabbits to reveal patterns of tick attachment. Of the 2426 Ixodes ticks collected from 42 rabbits, larvae were by far the most abundant life stage (2360), peaking in abundance in autumn, while small numbers of nymphs (62) and adults (4) were present in winter and summer respectively. Larvae were found all over the body, whereas adults and nymphs were predominantly attached to the head, suggesting that the mature life stages use the host landscape differently, or that adults or nymphs may be groomed off the body. The most abundant tick species, as determined by morphology and DNA sequencing, was Ixodes holocyclus, a generalist tick responsible for significant human and companion animal health concerns in Australia. Our findings highlight the importance of understanding the role of introduced wildlife in tick dynamics particularly in novel ecosystems where non-native hosts may be more abundant than native hosts.

Description of a new species of Ixodes (Acari: Ixodidae) and first report of Ixodes lasallei and Ixodes bocatorensis in Brazil.

In a recent study, we relegated the taxon Ixodes aragaoi Fonseca, 1935 to a junior synonym of Ixodes fuscipes Koch, 1844, and reinstated Ixodes spinosusNeumann, 1899 (a former synonym of I. fuscipes) to a valid species. We examined all lots of ticks formerly identified as I. fuscipes or I. aragaoi in three tick collections of Brazil. Through morphological analysis, some of the examined specimens could not be assigned to either I. fuscipes or I. spinosus based on the examination of the type specimens of these two species. Herein, we report these ticks to represent three different species: Ixodes catarinensis n. sp. Onofrio & Labruna, Ixodes lasalleiMéndez Arocha and Ortiz, 1958, and Ixodes bocatorensisApanaskevich and Bermúdez, 2017. The latter two species are reported for the first time in Brazil. Phylogenetic analysis inferred from the tick 16S rRNA gene partial sequences corroborated our morphological analysis, indicating that I. spinosus, I. lasallei, I. bocatorensis, and I. catarinensis n. sp. form a natural group of neotropical ticks. With the present study, the number of Ixodes species in Brazil increases from 9 to 12. We propose a new identification key for females and males of Ixodes species currently recognized in Brazil.

Tick burden on European roe deer (Capreolus capreolus) from Saxony, Germany, and detection of tick-borne encephalitis virus in attached ticks.

Southern Germany is known as tick-borne encephalitis (TBE) risk area; however, the north of the country is almost free of human TBE cases. Due to its location in the transition zone between TBE risk areas and areas with only sporadic cases, Saxony is of importance in the surveillance of TBE. Roe deer (Capreolus capreolus), showing high seroprevalence of TBE virus (TBEV) antibodies, are considered to be sentinels for TBE risk assessment. Thus, roe deer could be used as indicators helping to better understand the focality of the TBEV in nature and as a possible source to isolate TBEV. Therefore, the aims of this study were to examine roe deer coats for the presence of ticks to establish the tick burden and to detect the TBEV in attached ticks. One hundred thirty-four roe deer coats were provided by hunters from the Hunting Association in Saxony (August 2017-January 2019). The coats were frozen at - 80 °C and after de-freezing examined on both sides-inside and outside. Attached and nonattached ticks were collected, morphologically identified and tested using real-time RT-PCR for the presence of TBEV. In total, 1279 ticks were found on 48 coats. The predominant species was Ixodes ricinus (99.76%; n = 1276). Three remaining specimens were Ixodes spp. (0.16%, 1 female and 1 nymph) and Dermacentor reticulatus (0.08%, 1 male). The average infestation rate was 26.7 (SD = 69.5), with maximum of 439 ticks per animal. Females were the dominant life stage of ticks (n = 536; 42%), followed by nymphs (n = 397; n = 31.1%), males (n = 175; 13.7%), and larvae (n = 168; 13.2%). Only half of collected ticks were attached (n = 662; 51.8%). TBEV was detected only in one tick out of 1279 tested ticks. It was a female infesting a roe deer from Saxon Switzerland-East Ore Mountain. The results show that the method used in this study is not sufficient as a sentinel marker to predict TBEV spreading in nature. Although previous studies demonstrated the usefulness of serological testing of roe deer in order to trace TBE-endemic regions, using ticks attached to them to get virus isolates is not productive.

Synonymy of Ixodes aragaoi with Ixodes fuscipes, and reinstatement of Ixodes spinosus (Acari: Ixodidae).

Ixodes fuscipes Koch, 1844, described from Brazil, has been considered a valid species with records from Brazil and Peru. Ixodes spinosus Neumann, 1899, also described from Brazil, has been considered a synonym of I. fuscipes. In 2014, Ixodes aragaoi Fonseca, 1935 was redescribed as a valid species for Brazil and Uruguay. Until the present study, one major difference between the females of I. fuscipes and I. aragaoi was the horn-like auriculae in the former versus the ridge-like auriculae in the later, but this morphological difference was not supported after examination of the holotype of I. fuscipes and a syntype female of I. spinosus. Surprisingly, we found the type of I. fuscipes to have ridge-like auriculae, in contrast to the horn-like auriculae of I. spinosus. Comparisons of the I. fuscipes holotype with the syntypes of I. aragaoi revealed that they correspond to the same species. Therefore, we redescribe I. fuscipes, relegate I. aragaoi to a junior synonym of I. fuscipes, and reinstate I. spinosus as a valid species. After examining all lots of I. fuscipes in four tick collections, no specimen was recognized as I. fuscipes when compared with the type specimen of this taxon. On the other hand, specimens previously identified as I. aragaoi are now confirmed as I. fuscipes, with bona fide records for Brazil and Uruguay. Some of the specimens previously reported as I. fuscipes are now confirmed as I. spinosus, with records in two Brazilian biomes, Amazon and Atlantic rainforest. We present lists of the ticks examined in this study, with their current taxonomic status.

The Ixodes ricinus complex (Acari: Ixodidae) in the Southern Cone of America: Ixodes pararicinus, Ixodes aragaoi, and Ixodes sp. cf. I. affinis.

The goal of this study was to clarify the taxonomic status of the Ixodes ricinus complex in the Southern Cone of America, by using morphological characters and molecular markers (mitochondrial 16SrDNA and cox1 genes). The morphological analysis indicates that three different taxa of the I. ricinus complex occur in this region: Ixodes pararicinus, Ixodes aragaoi, and Ixodes sp. cf. I. affinis. The most prominent diagnostic character among them is the size of scutal punctations in both male and female ticks. In the males of Ixodes sp. cf. I. affinis, the punctations on the central field and along the median marginal groove of the scutum are clearly larger than in the males of I. aragaoi and I. pararicinus, while the punctations of I. aragaoi are larger but less numerous than in I. pararicinus. The punctations in Ixodes sp. cf. I. affinis females are larger and deeper than in females of I. aragaoi and I. pararicinus, and those of I. aragaoi are slightly larger than in I. pararicinus. The length of the lateral posterior denticles of the male hypostome is comparatively longer in I. aragaoi than in the other two species, and longer in Ixodes sp. cf. I. affinis than in I. pararicinus. In the 16S analysis, I. pararicinus and I. aragaoi are monophyletic (99% and 98% bootstrap support, respectively), while Ixodes cf. I. affinis does not represent a single lineage. In the cox1 analysis, both I. pararicinus and I. aragaoi are well-defined taxa, but the bootstrap support for Ixodes sp. cf. I. affinis is low (67%). In general, there are considerable 16SrRNA differences among lineages of Ixodes sp. cf. I. affinis from different geographical areas. These results may be indicative of the existence of different species. The populations morphologically compatible with I. affinis from Argentina, Colombia, Panama, Belize, and USA should be provisionally named as Ixodes sp. cf. I. affinis until an integrative taxonomic work with further evidence redefines whether or not this taxon actually represents a species complex.