Range Expansion of Native and Invasive Ticks: A Looming Public Health Threat.

Native and invasive tick species pose a serious public health concern in the United States. Range expansion of several medically important tick species has resulted in an increasing number of communities at risk for exposure to ticks and tick-borne pathogens.

A case of morphological anomalies in Amblyomma americanum (Acari: Ixodidae) collected from nature.

We describe a case of morphological anomalies in Amblyomma americanum, a medically important species associated with several human diseases and medical conditions. Based on morphological characters using dichotomous morphological keys, high-resolution light microscopy, and scanning electron microscopy imaging, the tick was identified as Am. americanum nymph exhibiting various morphological anomalies including ectromely associate with asymmetry, olygomely (lack) of the fourth left leg, and schizomely (bifurcation of palpus) on the right side. We believe this is the first report of the presence of several spontaneous anomalies in one Am. americanum specimen. Morphological identity of the specimen was corroborated by DNA sequencing of the mitochondrial 16S region. We discuss recent reports of morphological anomalies in ixodid ticks and emphasize the significance of additional studies of teratology in medically important tick species and its potential implications.

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.

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.

Enhancement of Risk for Lyme Disease by Landscape Connectivity, New York, New York, USA.

Most tickborne disease studies in the United States are conducted in low-intensity residential development and forested areas, leaving much unknown about urban infection risks. To understand Lyme disease risk in New York, New York, USA, we conducted tick surveys in 24 parks throughout all 5 boroughs and assessed how park connectivity and landscape composition contribute to Ixodes scapularis tick nymphal densities and Borrelia burgdorferi infection. We used circuit theory models to determine how parks differentially maintain landscape connectivity for white-tailed deer, the reproductive host for I. scapularis ticks. We found forested parks with vegetated buffers and increased connectivity had higher nymph densities, and the degree of park connectivity strongly determined B. burgdorferi nymphal infection prevalence. Our study challenges the perspective that tickborne disease risk is restricted to suburban and natural settings and emphasizes the need to understand how green space design affects vector and host communities in areas of emerging urban tickborne disease.

Host-Feeding Behavior of Mosquitoes in the Florida Everglades.

Background: West Nile virus (WNV), Everglades virus (EVEV), and five species of Orthobunyavirus were isolated from mosquitoes collected in the Everglades in 2016-2017. Prior studies of blood meals of mosquitoes in southern Florida have related findings to acquisition and transmission of EVEV, St. Louis encephalitis virus, and WNV, but not the Orthobunyavirus viruses associated with the subgenus Melanoconion of the genus Culex. Materials and Methods: In the present study, blood-fed mosquitoes were collected in the Everglades in 2016, 2017, 2021, and 2022, and from an industrial site in Naples, FL in 2017. Blood meals were identified to host species by PCR assays using mitochondrial cytochrome b gene. Results: Blood meals were identified from Anopheles crucians complex and 11 mosquito species captured in the Florida Everglades and from 3 species collected from an industrial site. The largest numbers of blood-fed specimens were from Culex nigripalpus, Culex erraticus, Culex cedecei, and Aedes taeniorhynchus. Cx. erraticus fed on mammals, birds, and reptiles, particularly American alligator. This mosquito species could transmit WNV to American alligator in the wild. Cx. nigripalpus acquired blood meals primarily from birds and mammals and frequently fed on medium-sized mammals and white-tailed deer. Water and wading birds were the primary avian hosts for Cx. nigripalpus and Cx. erraticus in the Everglades. Wading birds are susceptible to WNV and could serve as reservoir hosts. Cx. cedecei fed on five species of rodents, particularly black and hispid cotton rats. EVEV and three different species of Orthobunyavirus have been isolated from the hispid cotton rat and Cx. cedecei in the Everglades. Cx. cedecei is likely acquiring and transmitting these viruses among hispid cotton rats and other rodents. The marsh rabbit was a frequent host for An. crucians complex. An. crucians complex, and other species could acquire Tensaw virus from rabbits. Conclusions: Our study contributes to a better understanding of the host and viral associations of mosquito species in southwestern Florida.

Prevalence of Dirofilaria immitis (Spirurida: Onchocercidae) infection in Aedes, Culex, and Culiseta mosquitoes from north San Joaquin Valley, CA.

Canine heartworm is one of the most serious infections primarily affecting domestic dogs but will also infect cats and wild canids. To evaluate the potential of mosquitoes as vectors of dog heartworm, Dirofilaria immitis (Leidy) in San Joaquin County, CA, we collected mosquitoes in 2011 and analyzed for infection with heartworm by using polymerase chain reaction. Of 3,000 mosquito pools (total number of specimens = 36,554), D. immitis DNA was detected in 97 pools of seven species, and the overall minimum infection rate (MIR) for all mosquito species was 2.69: Culex pipiens L. (n = 40; MIR = 3.66), Culex tarsalis Coquillett (n = 25; MIR = 1.89), Culiseta incidens (Thomson) (n = 11; MIR = 2.81), Aedes vexans (Meigen) (n = 7; MIR = 2.18), Aedes melanimon Dyar (n = 5; MIR = 4.64), Culex erythrothorax Dyar (n = 5; MIR = 3.96), and Culiseta inornata (Williston) (n = 4; MIR = 2.65). Cx. pipiens and Cx. tarsalis had the highest number of D. immitis infections and collectively accounted for 67% of all positive pools. Ae. melanimon, Ae. vexans, and Cx. erythrothorax were found to be infected with D. immitis only in rural and agriculture areas, whereas infections in other species were identified in rural and agriculture areas, and urban and residential settings. The majority of positive pools were identified from June through November and peaked during August through October. This is the first report of D. immitis infection in Ae. melanimon, Cx. erythrothorax, Cx. tarsalis, Cs. incidens, and Cs. inornata. The frequent detection of D. immitis in field-collected Cx. pipiens and Cx. tarsalis in concert with their seasonal abundance and widespread distribution suggest a central role for these species in dog heartworm transmission. Other species, including Ae. vexans, Ae. melanimon, Cs. incidens, Cs. inornata, and Cx. erythrothorax, may play a secondary role in transmission.

Evidence of Uranotaenia sapphirina (Diptera: Culicidae) feeding on annelid worms in the Northeastern United States.

Mosquito host-feeding behavior is an important parameter for determining the vector potential of mosquito species in a given locale. Despite the recent discovery of Uranotaenia sapphirina Osten Sacken feeding on annelid hosts in Florida, host association studies for this mosquito species in the United States remain limited. To investigate the blood-feeding pattern of Ur. sapphirina in the northeastern United States, mosquitoes were collected from Massachusetts, Connecticut, and New Jersey using CDC miniature light traps, peat fiber resting boxes, gravid traps, and backpack aspirators. Vertebrate and invertebrate hosts of this mosquito species were identified through PCR amplification and nucleotide sequencing of portions of the mitochondrial cytochrome b gene and the 28S ribosomal RNA gene, respectively. Of 21 (24.7%) specimens successfully identified to host species, 47.6% contained solely annelid blood, 14.3% mammalian blood, 14.3% avian blood, and 23.8% with mixed blood of annelid and avian origin. The mud earthworm, Sparganophilus tennesseensis Reynolds (Haplotaxida: Sparganophilidae), was identified as the most common host (n = 14, including mixed bloods), followed by American robin, Turdus migratorius (n = 7, including mixed bloods). Testing of these blood engorged mosquitoes for West Nile virus and eastern equine encephalitis virus did not result in any positive specimens. This is the first report of Ur. sapphirina feeding on annelids and on both vertebrate and invertebrate hosts in mixed bloodmeals in the northeastern United States. Our findings support the recent report of Ur. sapphirina feeding on invertebrates and further emphasizes the inclination of some mosquito species to feed on a wider range of hosts spanning nontraditional taxonomic groups.

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.

EVIDENCE OF PROTOZOAN AND BACTERIAL INFECTION AND CO-INFECTION AND PARTIAL BLOOD FEEDING IN THE INVASIVE TICK HAEMAPHYSALIS LONGICORNIS IN PENNSYLVANIA.

The Asian longhorned tick, Haemaphysalis longicornis, an invasive tick species in the United States, has been found actively host-seeking while infected with several human pathogens. Recent work has recovered large numbers of partially engorged, host-seeking H. longicornis, which together with infection findings raises the question of whether such ticks can reattach to a host and transmit pathogens while taking additional bloodmeals. Here we conducted molecular blood meal analysis in tandem with pathogen screening of partially engorged, host-seeking H. longicornis to identify feeding sources and more inclusively characterize acarological risk. Active, statewide surveillance in Pennsylvania from 2020 to 2021 resulted in the recovery of 22/1,425 (1.5%) partially engorged, host-seeking nymphal and 5/163 (3.1%) female H. longicornis. Pathogen testing of engorged nymphs detected 2 specimens positive for Borrelia burgdorferi sensu lato, 2 for Babesia microti, and 1 co-infected with Bo. burgdorferi s.l. and Ba. microti. No female specimens tested positive for pathogens. Conventional PCR blood meal analysis of H. longicornis nymphs detected avian and mammalian hosts in 3 and 18 specimens, respectively. Mammalian blood was detected in all H. longicornis female specimens. Only 2 H. longicornis nymphs produced viable sequencing results and were determined to have fed on black-crowned night heron, Nycticorax nycticorax. These data are the first to molecularly confirm H. longicornis partial blood meals from vertebrate hosts and Ba. microti infection and co-infection with Bo. burgdorferi s.l. in host-seeking specimens in the United States, and the data help characterize important determinants indirectly affecting vectorial capacity. Repeated blood meals within a life stage by pathogen-infected ticks suggest that an understanding of the vector potential of invasive H. longicornis populations may be incomplete without data on their natural host-seeking behaviors and blood-feeding patterns in nature.

Comparison of acarological risk metrics derived from active and passive surveillance and their concordance with tick-borne disease incidence.

Tick-borne diseases continue to threaten human health across the United States. Both active and passive tick surveillance can complement human case surveillance, providing spatio-temporal information on when and where humans are at risk for encounters with ticks and tick-borne pathogens. However, little work has been done to assess the concordance of the acarological risk metrics from each surveillance method. We used data on Ixodes scapularis and its associated human pathogens from Connecticut (2019-2021) collected through active collections (drag sampling) or passive submissions from the public to compare county estimates of tick and pathogen presence, infection prevalence, and tick abundance by life stage. Between the surveillance strategies, we found complete agreement in estimates of tick and pathogen presence, high concordance in infection prevalence estimates for Anaplasma phagocytophilum, Borrelia burgdorferi sensu stricto, and Babesia microti, but no consistent relationships between actively and passively derived estimates of tick abundance or abundance of infected ticks by life stage. We also compared nymphal metrics (i.e., pathogen prevalence in nymphs, nymphal abundance, and abundance of infected nymphs) with reported incidence of Lyme disease, anaplasmosis, and babesiosis, but did not find any consistent relationships with any of these metrics. The small spatial and temporal scale for which we had consistently collected active and passive data limited our ability to find significant relationships. Findings are likely to differ if examined across a broader spatial or temporal coverage with greater variation in acarological and epidemiological outcomes. Our results indicate similar outcomes between some actively and passively derived tick surveillance metrics (tick and pathogen presence, pathogen prevalence), but comparisons were variable for abundance estimates.

Vector host-feeding preferences drive transmission of multi-host pathogens: West Nile virus as a model system.

Seasonal epizootics of vector-borne pathogens infecting multiple species are ecologically complex and difficult to forecast. Pathogen transmission potential within the host community is determined by the relative abilities of host species to maintain and transmit the pathogen and by ecological factors influencing contact rates between hosts and vectors. Increasing evidence of strong feeding preferences by a number of vectors suggests that the host community experienced by the pathogen may be very different from the local host community. We developed an empirically informed transmission model for West Nile virus (WNV) in four sites using one vector species (Culex pipiens) and preferred and non-preferred avian hosts. We measured strong feeding preferences for American robins (Turdus migratorius) by Cx. pipiens, quantified as the proportion of Cx. pipiens blood meals from robins in relation to their abundance (feeding index). The model accurately predicted WNV prevalence in Cx. pipiens at three of four sites. Sensitivity analysis revealed feeding preference was the most influential parameter on intensity and timing of peak WNV infection in Cx. pipiens and a threshold feeding index for transmission was identified. Our findings indicate host preference-induced contact heterogeneity is a key mediator of vector-borne pathogen epizootics in multi-species host communities, and should be incorporated into multi-host transmission models.

Vector-host interactions of Culex pipiens complex in northeastern and southwestern USA.

Studies on the vector-host interactions of Culex pipiens complex mosquitoes by sequencing portions of mitochondrial cytochrome b gene indicate that Cx. p. pipiens f. pipiens predominantly feed on avian hosts (93.1%), and focus feeding activity on several key bird species, in particular the American robin, the gray catbird, and the house sparrow in Connecticut. However, Cx. p. quinquefasciatus indiscriminately feed on both birds and mammals. Culex p. quinquefasciatus in Harris County - Texas and southern California acquired 39.1% and 88.2% of bloodmeals from birds, respectively. Mammalian-derived bloodmeals constituted 52.5% and 9.6% in the two regions, respectively. The most frequent avian hosts for this mosquito species in the southwestern U.S. were the mourning dove, the white-winged dove, the house sparrow and the house finch. Humans infrequently served as the source of bloodmeals for Cx. p. pipiens and Cx. p. quinquefasciatus. Microsatellite analysis of mosquitoes from Chicago, Illinois showed that Cx. p. pipiens f. pipiens with mammalian- derived bloodmeals had significantly higher ancestry and proportion of hybrids from Cx. p. pipiens f. molestus than did those with avian-derived bloodmeals.

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.

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.

Spatiotemporal distribution, abundance, and host interactions of two invasive vectors of arboviruses, Aedes albopictus and Aedes japonicus, in Pennsylvania, USA.

BACKGROUND: Aedes albopictus and Aedes japonicus, two invasive mosquito species in the United States, are implicated in the transmission of arboviruses. Studies have shown interactions of these two mosquito species with a variety of vertebrate hosts; however, regional differences exist and may influence their contribution to arbovirus transmission. METHODS: We investigated the distribution, abundance, host interactions, and West Nile virus infection prevalence of Ae. albopictus and Ae. japonicus by examining Pennsylvania mosquito and arbovirus surveillance data for the period between 2010 and 2018. Mosquitoes were primarily collected using gravid traps and BG-Sentinel traps, and sources of blood meals were determined by analyzing mitochondrial cytochrome b gene sequences amplified in PCR assays. RESULTS: A total of 10,878,727 female mosquitoes representing 51 species were collected in Pennsylvania over the 9-year study period, with Ae. albopictus and Ae. japonicus representing 4.06% and 3.02% of all collected mosquitoes, respectively. Aedes albopictus was distributed in 39 counties and Ae. japonicus in all 67 counties, and the abundance of these species increased between 2010 and 2018. Models suggested an increase in the spatial extent of Ae. albopictus during the study period, while that of Ae. japonicus remained unchanged. We found a differential association between the abundance of the two mosquito species and environmental conditions, percent development, and median household income. Of 110 Ae. albopictus and 97 Ae. japonicus blood meals successfully identified to species level, 98% and 100% were derived from mammalian hosts, respectively. Among 12 mammalian species, domestic cats, humans, and white-tailed deer served as the most frequent hosts for the two mosquito species. A limited number of Ae. albopictus acquired blood meals from avian hosts solely or in mixed blood meals. West Nile virus was detected in 31 pools (n = 3582 total number of pools) of Ae. albopictus and 12 pools (n = 977 total pools) of Ae. japonicus. CONCLUSIONS: Extensive distribution, high abundance, and frequent interactions with mammalian hosts suggest potential involvement of Ae. albopictus and Ae. japonicus in the transmission of human arboviruses including Cache Valley, Jamestown Canyon, La Crosse, dengue, chikungunya, and Zika should any of these viruses become prevalent in Pennsylvania. Limited interaction with avian hosts suggests that Ae. albopictus might occasionally be involved in transmission of arboviruses such as West Nile in the region.

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.

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.

Established Population of the Gulf Coast Tick, Amblyomma maculatum (Acari: Ixodidae), Infected with Rickettsia parkeri (Rickettsiales: Rickettsiaceae), in Connecticut.

We identified an established population of the Gulf Coast tick (Amblyomma maculatum Koch) infected with Rickettsia parkeri in Connecticut, representing the northernmost range limit of this medically relevant tick species. Our finding highlights the importance of tick surveillance and public health challenges posed by geographic expansion of tick vectors and their pathogens.