Introduction of the ectoparasite Rhipicephalus pulchellus (Ixodida: Ixodidae) into Connecticut with a human traveler from Tanzania, and a review of its importation records into the United States.

Globalization, increased frequency of travel, and a rise in legal and illegal animal trades can introduce exotic ticks into the United States. We herein report the importation of a male Rhipicephalus pulchellus (Gerstäcker) on a human traveler returning to Connecticut from Tanzania, Africa, and review historical importation records of this species into the United States. This common tick is broadly distributed throughout East Africa, from Eritrea to Tanzania, has a wide host range on domestic animals and wildlife, and has been most frequently introduced into the United States on tick-infested wild animal hosts and animal trophies, but documentation of importation on humans has been rare. Archival records at the United States Department of Agriculture's National Veterinary Services Laboratories show R. pulchellus has been introduced into the United States at least 40 times over the last 62 yr. Rhipicephalus pulchellus has been linked to Rickettsia conorii, the agent of boutonneuse fever in humans, Crimean-Congo hemorrhagic fever orthonairovirus, and Nairobi sheep disease orthonairovirus. Given the potential for this exotic tick to introduce animal or human pathogens, proper surveillance, interception, identification, and reporting of these ticks are vital in protecting human and animal health.

Orally delivered fipronil-laced bait reduces juvenile blacklegged tick (Ixodes scapularis) burdens on wild white-footed mice (Peromyscus leucopus).

While the topical acaricidal treatment of rodent pathogen reservoirs has been readily explored over the past several decades, oral systemic acaricidal treatment is only recently gaining traction as an alternative approach to the management of ticks and tick-borne pathogens. Recent laboratory tests have shown promise in the effectiveness of this systemic strategy against the blacklegged tick (Ixodes scapularis) and a Canadian field evaluation was recently published, but no recent field data from the United States yet exist. With this research, we sought to field deploy a commercially available fipronil-laced bait (Kaput® Flea Control Bait, Scimetrics LLC., Wellington, CO, USA; 0.005% fipronil; Environmental Protection Agency Reg. No. 72500-28), in an alternate use targeting white-footed mice (Peromyscus leucopus) to determine bait acceptance and potential impacts to juvenile I. scapularis burdens. Bait was readily accepted by wild P. leucopus and other rodent reservoirs. An ad libitum distribution strategy as well as placing smaller volumes of fipronil-laced bait within individual Sherman traps both resulted in significant reductions (57-94%) in juvenile I. scapularis burdens as compared to control over two years. The oral delivery of systemic acaricides shows promise in reduction of I. scapularis burdens on P. leucopus and should be further explored to determine effectiveness on host-seeking tick abundances, associated pathogen infection, and potentially incorporated into integrated tick management programs.

Anomalous morphologies in Ixodes scapularis feeding on human hosts.

Cases of anomalous morphologies in the blacklegged tick, Ixodes scapularis, have been reported in both field-collected and human-biting specimen in the Northeastern and Midwestern United States, complicating the identification of this medically important tick species. We herein describe four cases of morphological anomalies in I. scapularis females exhibiting nanism and abnormally small genital apertures. We also report a female I. scapularis displaying slight asymmetry in the lower abdomen oriented toward the right side and an abnormal anal groove completely enclosing the anus. The identity of each specimen was confirmed using taxonomic keys, high resolution light and scanning electron microscopy imaging, and DNA sequencing of the 18S rRNA gene. All specimens described in this study were found parasitizing human hosts and were submitted to the Connecticut Agricultural Experiment Station-Tick Testing Laboratory in 2021 for species identification and pathogen screening. Here, we also discuss recent reports of teratological abnormalities in I. scapularis as well as likely causes for such deformities and potential implications.

Rhipicephalus capensis (Acari: Ixodidae), A geographically restricted South African tick, returning with a human traveler to the United States.

Accelerated frequency of recreational travel, globalization of business, and legal and illegal plant and animal trades have contributed to enduring introductions of exotic ticks into the United States. We herein report a new incursion of a female Rhipicephalus capensis on a human traveler returning to Connecticut from South Africa. Natural distribution of R. capensis is restricted to the Western Cape Province and southwestern portion of the Northern Cape Province of South Africa, an area called the Fynbos Biome, and adults of this species primarily parasitize large, wild ungulate hosts. Only one previous international introduction of this tick is documented on imported South African plant material into the United States in 1985. The specimen described here was identified initially by morphological means and subsequently, a partial DNA sequence for the mitochondrial ribosomal RNA gene was generated in a PCR assay, which showed 94.86% identity to an R. capensis sequence in GenBank. We also provide information on several other previously unreported or under-reported incursions by South African ticks into the United States in association with imported Fynbos floricultural materials and speckled Cape tortoises, Chersobius signatus. Documentation of these additional exotic tick species incursions highlights ongoing challenges of the international movement of humans, animals, and other goods carrying ticks of human and veterinary importance.

Integrated Tick Management in Guilford, CT: Fipronil-Based Rodent-Targeted Bait Box Deployment Configuration and Peromyscus leucopus (Rodentia: Cricetidae) Abundance Drive Reduction in Tick Burdens.

Integrated tick management (ITM) is a comprehensive strategy used to reduce presence of ticks and their associated pathogens. Such strategies typically employ a combination of host and non-host targeted treatments which often include fipronil-based, rodent-targeted bait boxes. Bait boxes target small-bodied rodents, specifically white-footed mice (Peromyscus leucopus Rafinesque) that not only play a crucial role in the blacklegged tick (Ixodes scapularis Say (Ixodida: Ixodidae)) life cycle, but also in the transmission of numerous pathogens, primarily Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner (Spirochaetales: Spirochaetaceae), the causal agent of Lyme disease. This study aimed to determine the effect of bait box deployment configuration on tick burden reduction while also further exploring bait consumption and P. leucopus abundances as measures of bait box usage and effectiveness. Boxes were deployed on nine properties within each of six neighborhoods (n = 54) in two different configurations: grid and perimeter. Multiple factors were analyzed as potential predictors for reduction in tick burdens using a backward stepwise selection procedure. Results confirmed the perimeter configuration was a more effective deployment strategy. In addition, overall P. leucopus abundance was a significant predictor of tick burden reduction while bait consumption was not. These findings not only further support the recommended perimeter deployment configuration but provide insight into effective utilization in areas of high P. leucopus abundance. The identification of this significant relationship, in addition to configuration, can be utilized by vector control professionals and homeowners to make informed decisions on bait box placement to make sustained impacts on the I. scapularis vector and associated pathogens within an ITM framework.

First Record of Established Populations of the Invasive Pathogen Vector and Ectoparasite Haemaphysalis longicornis (Acari: Ixodidae) in Connecticut, United States.

A number of invasive tick species capable of transmitting pathogens have been accidentally introduced into the U.S. in recent years. The invasion and further range expansion of these exotic ticks have been greatly facilitated by frequent global travel and trade as well as increases in legal and illegal importation of animals. We describe the discovery of the first established populations of Haemaphysalis longicornis Neumann and the first fully engorged human parasitizing specimen documented through passive tick surveillance in Fairfield County, Connecticut, U.S. We also report several individual specimens of this invasive arthropod and vector of multiple pathogens of medical and veterinary importance collected through active tick surveillance from three counties (Fairfield, New Haven, and New London). Considering the potential for invasive ticks to transmit numerous native and emerging pathogens, the implementation of comprehensive surveillance programs will aid in prompt interception of these ticks and reduce the risk of infection in humans and wildlife.

Effective control of the motile stages of Amblyomma americanum and reduced Ehrlichia spp. prevalence in adults via permethrin treatment of white-tailed deer in coastal Connecticut, USA.

The lone star tick, Amblyomma americanum, is a common human-biting species whose range has been largely restricted to the southeastern United States, until recent detections of established populations on Long Island, New York and throughout coastal southern New England. We evaluated the effectiveness of topical treatment of 10 % permethrin delivered via 4-poster devices to white-tailed deer, Odocoileus virginianus, in the management of a newly discovered A. americanum population in Norwalk, Connecticut. Using a high-density deployment of one 4-poster device/12.7 ha, we were successful in significantly reducing densities of host-seeking adults (93 % reduction), nymphs (92 %), and larvae (96 %) from 2018 to 2020. We also documented a significant reduction (87 %) in parasitizing adults and nymphs on white-tailed deer from 2018 to 2019. The prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii combined in host-seeking adults declined significantly from 47 % at the time the A. americanum population was discovered in 2017 to 7% in 2020. However, the prevalence in nymphs remained static (∼9%) throughout the study period. These data demonstrate that, when properly deployed in a density-dependent manner in terms of deer abundance, 4-poster devices can effectively manage parasitizing and host-seeking A. americanum populations and reduce the prevalence of two ehrlichial species of public health importance.

Rabbit Bot Fly Furuncular, Tracheopulmonary, and Human Bot Fly Infestations in Connecticut (Oestridae: Cuterebrinae).

Endemic and tropical human bot infestations are relatively uncommon or unreported in the United States. We report two cases in Connecticut: an unusual furuncular and respiratory myiasis by the rabbit bot Cuterebra buccata (Fab.) (Diptera: Oestridae) in a 74-yr-old male and a case of human bot fly, Dermatobia hominis (L.) (Diptera: Oestridae), myiasis in a 4-yr-old female with a tropical travel history with her family. Identification of C. buccata was based morphologically, in part, on spinal armature and further corroborated by DNA sequencing of the mitochondrial COI gene and comparison to the National Center for Biotechnology Information GenBank DNA sequence database. The resulting annotated sequence data were deposited into the National Center for Biotechnology Information GenBank. The unique medical aspects, and limitations and specifics on bot fly larval habits and identification are discussed.

Barriers to Effective Tick Management and Tick-Bite Prevention in the United States (Acari: Ixodidae).

Lyme and other tick-borne diseases are increasing in the United States. Development of tick control tools have focused primarily on the blacklegged tick, Ixodes scapularis Say. Application of acaricides or entomopathogenic fungal agents to kill host-seeking ticks or ticks on rodents can suppress I. scapularis abundance in residential landscapes, but evidence is lacking for impact on human tick bites or tick-borne disease. Similar studies remain limited for the lone star tick, Amblyomma americanum (L.). Other knowledge gaps include how well homeowners and pest control companies perform in the broadcast application of tick-killing products, relative to high efficacy reported in research studies, and the tick-killing potential of natural product formulations exempt from Environmental Protection Agency registration. Area-wide control based on preventing ticks from feeding on their main reproductive host, the white-tailed deer, can suppress populations of both I. scapularis and A. americanum. Some studies also suggest an impact on Lyme disease cases, but this needs to be further validated in larger-scale intervention studies. The effectiveness, scale, cost, and implementation of various tick management strategies are important considerations in efforts to reduce human tick encounters and tick-borne disease. Additional barriers include weak incentives for industry and academia to develop, test, and register new tick and pathogen control technologies, including vaccines targeting humans, tick reproductive hosts, or wildlife pathogen reservoirs. Solutions will need to be 'two-pronged': improving the tick and pathogen control toolbox and strengthening the public health workforce engaging in tick control at local and state levels.

Enduring Challenge of Invasive Ticks: Introduction of Amblyomma oblongoguttatum (Acari: Ixodidae) into the United States on a Human Traveler Returning from Central America.

Introduction of exotic tick vectors of bacteria, protozoa, viruses, and filarial parasites into the United States has accelerated in recent years, primarily because of globalization, increased frequency of travel, and a rise in legal and illegal animal trades. We herein report introduction of a live specimen of Amblyomma oblongoguttatum on a human into the United States from Central America, and we review 4 previous similar incidents. This tick species occurs widely in the neotropics, from western and southern Mexico, southwards through Central America, to the northern half of South America. It is a potential vector of bacterial agents of spotted fever group rickettsioses, raising concern that if A. oblongoguttatum ticks become established in this country, they might also be able to carry pathogens of human and veterinary concern. Given the potential for exotic ticks as vectors of numerous pathogens, proper surveillance, interception, and identification of these ticks are vital to protecting human and veterinary health. Rigorous governmental inspections of imported livestock and pet animals at ports of entry and educating human travelers and medical practitioners about the risks should be part of an overall national tick program.

Humane Use of Cardiac Puncture for Non-Terminal Phlebotomy of Wild-Caught and Released Peromyscus spp.

The cardiac puncture technique for obtaining relatively large volume (50-150 µL) blood samples from sedated rodents has been used in research for nearly a century. Historically, its use to phlebotomize and then release live rodents was more common. However, recently its use in a non-terminal capacity frequently imparts negative connotations in part because exsanguination of sedated animals via cardiac puncture is now an American Veterinary Medical Association-approved euthanasia technique. This association has resulted in ethical concerns by manuscript reviewers and in a few instances, outright refusal by some peer-reviewed journals to publish research that utilized the technique. To counter the perceived negative associations with its non-terminal use, we summarized nearly two decades (2001-2019) of capture and handling data throughout Connecticut, resulting in over 7000 cardiac punctures performed on nearly 5000 sedated, live-captured and released Peromyscus spp. We show that our total handling mortality rate (3.7%) was comparable, if not lower, than similar field studies that utilized other phlebotomy techniques. Many public health, integrated tick management, and vector-borne disease ecology studies require samples from individual wild-caught Peromyscus spp. over time to determine intervention efficacy and pathogen infection monitoring, and in such field studies, post-operative care is not an option. Proper execution of cardiac puncture does not increase susceptibility of individuals to predation upon release as can potential ocular abnormalities or infections that can occur as the result of use of other techniques. We posit that neither exsanguination nor resulting euthanasia are requirements of cardiac puncture and that its use is entirely appropriate for obtaining blood samples from live-captured and released Peromyscus spp. Properly performed cardiac puncture is an excellent technique to obtain blood samples from sedated, individual Peromyscus spp. on multiple appropriately-spaced occasions over single trapping seasons while keeping animal welfare a top priority.

Administration of an Orally Delivered Substrate Targeting a Mammalian Zoonotic Pathogen Reservoir Population: Novel Application and Biomarker Analysis.

Reservoir-targeted vaccines (RTVs) have the potential to be effective at breaking the transmission cycle of many tick-borne pathogens including, but not limited to, Borrelia burgdorferi, B. miyamotoi, B. mayonii, Babesia microti, and Anaplasma phagocytophilum. To determine what proportion of a wild reservoir species we could effectively target, we distributed an experimental non-RTV Rhodamine B (RhB)-coated pellet formulation devoid of nutrient supplementation using bait boxes with ad libitum access, in battery-operated time-release bait stations, and by hand broadcast. Regardless of distribution method, a total of 208 of 242 (86%) white-footed mouse (Peromyscus leucopus) captures were positive for RhB by either pelage staining or by detecting fluorescent expression in vibrissae under a microscope. In bait box locations, 91% of captured mice were RhB-positive, 89% in hand broadcast locations, and 80% in time-release station locations. Based on results, we are confident that the bait formulation was readily accepted regardless of distribution technique, reached a substantial proportion of the reservoir population, and provides an effective vehicle to deliver a range of RTVs to targeted, wild, pathogen reservoir populations.

Field evaluation of a novel oral reservoir-targeted vaccine against Borrelia burgdorferi utilizing an inactivated whole-cell bacterial antigen expression vehicle.

Blacklegged ticks (Ixodes scapularis) are the principal vector for Borrelia burgdorferi, among other infectious agents, in the northeastern, mid-Atlantic, and upper midwestern USA. White-footed mice (Peromyscus leucopus) are the primary and most competent reservoir host of B. burgdorferi in the Northeast. Live reservoir-targeted vaccines (RTVs) to limit enzootic transmission of B. burgdorferi were previously developed and successfully evaluated in laboratory and controlled field trials. A novel, inactivated RTV was developed to minimize regulatory and market challenges facing previous RTVs based on live bacterial or viral vehicles. Thirty-two residential properties in Redding, Connecticut, participated in a field trial of an orally delivered, inactivated RTV efficacy study (2015-2016). During the two-year vaccination period, a significant decrease in the percentage of B. burgdorferi-infected I. scapularis larvae parasitizing P. leucopus was observed, as was a significant reduction in the percentage of infected P. leucopus on RTV-treated properties when compared to control properties. This novel inactivated RTV was effective in reducing numbers of B. burgdorferi-infected I. scapularis and B. burgdorferi-infected P. leucopus on properties where it was distributed.

Evaluating the effectiveness of an integrated tick management approach on multiple pathogen infection in Ixodes scapularis questing nymphs and larvae parasitizing white-footed mice.

We investigated the effectiveness of integrated tick management (ITM) approaches in reducing the burden of infection with Borrelia burgdorferi, Babesia microti, and Anaplasma phagocytophilum in Ixodes scapularis. We found a 52% reduction in encountering a questing nymph in the Metarhizium anisopliae (Met52) and fipronil rodent bait box treatment combination as well as a 51% reduction in the combined white-tailed deer (Odocoileus virginianus) removal, Met52, and fipronil rodent bait box treatment compared to the control treatment. The Met52 and fipronil rodent bait box treatment combination reduced the encounter potential with a questing nymph infected with any pathogen by 53%. Compared to the control treatment, the odds of collecting a parasitizing I. scapularis infected with any pathogen from a white-footed mouse (Peromyscus leucopus) was reduced by 90% in the combined deer removal, Met52, and fipronil rodent bait box treatment and by 93% in the Met52 and fipronil rodent bait box treatment combination. Our study highlights the utility of these ITM measures in reducing both the abundance of juvenile I. scapularis and infection with the aforementioned pathogens.

A seven-legged tick: Report of a morphological anomaly in Ixodes scapularis (Acari: Ixodidae) biting a human host from the Northeastern United States.

Cases of morphological anomalies in the blacklegged tick, Ixodes scapularis (Acari: Ixodidae), have recently been reported from the Northeastern and upper Midwestern United States, potentially complicating identification of this important vector of human disease-causing pathogens. We hereby report a case of a morphological anomaly in I. scapularis, biting a human host residing in Norwich, Connecticut. Using a dichotomous morphological key, high-resolution and scanning electron microscopy images, as well as DNA sequencing, the tick was identified as an adult female I. scapularis with three legs on the left side of the abdomen versus four on the right side, which we believe is the first case of ectromely in an adult I. scapularis. Using diagnostic genes in polymerase chain reaction, the specimen tested positive for Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum, the causative agents for Lyme disease and anaplasmosis, respectively, and also showed evidence of a rickettsial endosymbiont. Here we discuss recent reports of morphological anomalies in I. scapularis, and emphasize the significance of additional studies of teratology in this important tick species and its potential implications.

Determining Effects of Winter Weather Conditions on Adult Amblyomma americanum (Acari: Ixodidae) Survival in Connecticut and Maine, USA.

The lone star tick (Amblyomma americanum L.) is native to the United States, with its primary range encompassing the Southeast and portions of the Midwest. It is an aggressive ectoparasite that actively seeks out hosts through detection of carbon dioxide and vibrations and can transfer ehrlichiosis-causing bacteria as well as a carbohydrate that causes alpha-gal syndrome (red meat allergy) in humans. It has become of increasing concern as its range has recently expanded into coastal regions of the Northeast. Historically, harsh northeastern winter weather conditions made these areas inhospitable for A. americanum survival, but a warming climate coupled with increased host availability seem to have facilitated their range expansion. We developed a study to observe the effects of weather conditions on adult A. americanum overwintering survival. The study was conducted over three years in Connecticut and Maine. Ground-level conditions were manipulated to determine the effects of differing combinations of natural insulative barriers (leaf litter and snow accumulation) on adult A. americanum survival. We determined that there was a significant difference in survival between the two states, between years in Maine, and between sexes within Connecticut. However, presence or absence of snow and/or leaf litter had no impact on survival. Overall, we found a positive correlation between mean hourly temperature and adult survival in Maine, where temperatures were consistently below freezing. The results of this study can be included in an adaptive, predictive analytic model to accommodate the expected fluctuations and range expansion of A. americanum that will most likely accompany an increase in temperatures throughout the Northeast.

Impacts of Deciduous Leaf Litter and Snow Presence on Nymphal Ixodes scapularis (Acari: Ixodidae) Overwintering Survival in Coastal New England, USA.

Blacklegged ticks (Ixodes scapularis Say) are the vector for pathogens that cause more cases of human disease than any other arthropod. Lyme disease is the most common, caused by the bacterial spirochete Borrelia burgdorferi (Johnson, Schmid, Hyde, Steigerwalt, and Brenner) in the northeastern United States. Further knowledge of seasonal effects on survival is important for management and modeling of both blacklegged ticks and tick-borne diseases. The focus of our study was on the impact of environmental factors on overwintering success of nymphal blacklegged ticks. In a three-year field study conducted in Connecticut and Maine, we determined that ground-level conditions play an important role in unfed nymphal overwintering survival. Ticks in plots where leaf litter and snow accumulation were unmanipulated had significantly greater survival compared to those where leaf litter was removed (p = 0.045) and where both leaf litter and snow were removed (p = 0.008). Additionally, we determined that the key overwintering predictors for nymphal blacklegged tick survival were the mean and mean minimum temperatures within a year. The findings of this research can be utilized in both small- and large-scale management of blacklegged ticks to potentially reduce the risk and occurrence of tick-borne diseases.

Predicting spatiotemporal patterns of Lyme disease incidence from passively collected surveillance data for Borrelia burgdorferi sensu lato-infected Ixodes scapularis ticks.

Lyme disease is the most prevalent vector-borne disease in the United States. Ixodes scapularis, commonly referred to as the blacklegged tick, is the primary vector of Lyme disease spirochetes, Borrelia burgdorferi sensu lato (s.l.), in the eastern United States. Connecticut has pervasive populations of I. scapularis and remains a hotspot for Lyme disease. A primary aim of this study was to determine if passively collected data on human-biting I. scapularis ticks in Connecticut could serve as a useful proxy for Lyme disease incidence based on the cases reported by the Connecticut Department of Public Health (CDPH). Data for human-biting I. scapularis ticks submitted to the Tick Testing Laboratory at the Connecticut Agricultural Experiment Station (CAES-TTL), and tested for infection with B. burgdorferi s.l., were used to estimate the rate of submitted nymphs, nymphal infection prevalence, and the rate of submitted infected nymphs. We assessed spatiotemporal patterns in tick-based measures and Lyme disease incidence with generalized linear and spatial models. In conjunction with land cover and household income data, we used generalized linear mixed effects models to examine the association between tick-based risk estimates and Lyme disease incidence. Between 2007 and 2017, the CAES-TTL received 26,116 I. scapularis tick submissions and the CDPH reported 23,423 Lyme disease cases. The rate of submitted nymphs, nymphal infection prevalence, the rate of submitted infected nymphs, and Lyme disease incidence all decreased over time during this eleven-year period. The rate of submitted nymphs, the rate of submitted infected nymphs, and Lyme disease incidence were spatially correlated, but nymphal infection prevalence was not. Using a mixed modeling approach to predict Lyme disease incidence and account for spatiotemporal structuring of the data, we found the best fitting tested model included a strong, positive association with the rate of submitted infected nymphs and a negative association with the percent of developed land for each county. We show that within counties, submissions of B. burgdorferi s.l. infected nymphs were strongly and positively associated with inter-annual variation in reported Lyme disease cases. Tick-based passive surveillance programs may be useful in providing independent measures of entomological risk, particularly in settings where Lyme disease case reporting practices change substantially over time.

Distribution and Establishment of the Lone Star Tick in Connecticut and Implications for Range Expansion and Public Health.

In the United States, the lone star tick, Amblyomma americanum (L.) (Acari: Ixodidae), is an aggressive southeastern species whose range has reportedly been steadily expanding northward. The number of A. americanum specimens submitted to the Tick Testing Laboratory (TTL) at the Connecticut Agricultural Experiment Station (CAES) increased by 58% from the period of 1996-2006 (n = 488) to 2007-2017 (n = 773), mainly from Fairfield County in the southwestern corner of the state. The greatest numbers of A. americanum submissions to the CAES-TTL were from the City of Norwalk and a few adjacent municipalities. We also report the discovery of a large infestation of adult and nymphal lone star ticks detected on a dead male white-tailed deer, Odocoileus virginianus (Zimmerman) (Artiodactyla: Cervidae), on Manresa Island, Norwalk, in June 2017, indicating a long established, undetected population along the southwestern coast. A sample of nymphal and adult host-seeking A. americanum collected July 2017 from Manresa Island were tested and a proportion were positive for Ehrlichia chaffeensis, Ehrlichia ewingii, and Anaplasma phagocytophilum. The A. americanum tick and its associated disease pathogens are expected to become an increasing public health concern in southern New England.