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.

Host Associations of Culex pipiens: A Two-Year Analysis of Bloodmeal Sources and Implications for Arboviral Transmission in Southeastern Virginia.

Understanding vector-host interactions is crucial for evaluating the role of mosquito species in enzootic cycling and epidemic/epizootic transmission of arboviruses, as well as assessing vertebrate host contributions to maintenance and amplification in different virus foci. To investigate blood-feeding pattern of Culex pipiens, engorged mosquitoes were collected on a weekly basis at 50 sites throughout Suffolk, Virginia, using Centers for Disease Control and Prevention miniature light traps, BG-Sentinel traps, and modified Reiter gravid traps. Vertebrate hosts of mosquitoes were identified by amplifying and sequencing portions of the mitochondrial cytochrome b gene. Of 281 Cx. pipiens bloodmeals successfully identified to species, 255 (90.7%) contained solely avian blood, 13 (4.6%) mammalian, 1 (0.4%) reptilian, and 12 (4.3%) both avian and mammalian blood. Nineteen avian species were identified as hosts for Cx. pipiens with American robin (n = 141, 55.3% of avian hosts) and northern cardinal (n = 57, 22.4%) as the most common hosts. More American robin feedings took place in areas of higher development. Three mammalian species were also identified as hosts for Cx. pipiens with Virginia opossum and domestic cat as the most common hosts in this class (each n = 6, 46.2% of mammalian hosts). There was no significant seasonal difference in the proportion of bloodmeals obtained from avian hosts, but there was a decrease in the proportion of bloodmeals from mammalian hosts from spring to fall. One engorged specimen of Cx. pipiens with Virginia opossum-derived bloodmeal tested positive for West Nile virus (WNV), and another with black-and-white warbler-derived bloodmeal tested positive for eastern equine encephalitis virus. Our findings, in conjunction with the results of vector competence studies and virus isolation from field-collected mosquitoes, lend additional support that Cx. pipiens serves as the principal enzootic vector and potential epizootic/epidemic vector of WNV in southeastern Virginia.

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.

Host interactions of Aedes albopictus, an invasive vector of arboviruses, in Virginia, USA.

BACKGROUND: As an invasive mosquito species in the United States, Aedes albopictus is a potential vector of arboviruses including dengue, chikungunya, and Zika, and may also be involved in occasional transmission of other arboviruses such as West Nile, Saint Louis encephalitis, eastern equine encephalitis, and La Crosse viruses. Aedes albopictus feeds on a wide variety of vertebrate hosts, wild and domestic, as well as humans. METHODOLOGY/PRINCIPAL FINDINGS: In order to investigate blood feeding patterns of Ae. albopictus, engorged specimens were collected from a variety of habitat types using the Centers for Disease Control and Prevention light traps, Biogents Sentinel 2 traps, and modified Reiter gravid traps in southeast Virginia. Sources of blood meals were determined by the analysis of mitochondrial cytochrome b gene sequences amplified in PCR assays. Our aims were to quantify degrees of Ae. albopictus interactions with vertebrate hosts as sources of blood meals, investigate arboviral infection status, assess the influence of key socioecological conditions on spatial variability in blood feeding, and investigate temporal differences in blood feeding by season. Analysis of 961 engorged specimens of Ae. albopictus sampled between 2017-2019 indicated that 96%, 4%, and less than 1% obtained blood meals from mammalian, reptilian, and avian hosts, respectively. Domestic cats were the most frequently identified (50.5%) hosts followed by Virginia opossums (17.1%), white-tailed deer (12.2%), and humans (7.3%), together representing 87.1% of all identified blood hosts. We found spatial patterns in blood feeding linked to socioecological conditions and seasonal shifts in Ae. albopictus blood feeding with implications for understanding human biting and disease risk. In Suffolk Virginia in areas of lower human development, the likelihood of human blood feeding increased as median household income increased and human blood feeding was more likely early in the season (May-June) compared to later (July-October). Screening of the head and thorax of engorged Ae. albopictus mosquitoes by cell culture and RT-PCR resulted in a single isolate of Potosi virus. CONCLUSION AND SIGNIFICANCE: Understanding mosquito-host interactions in nature is vital for evaluating vectorial capacity of mosquitoes. These interactions with competent reservoir hosts support transmission, maintenance, and amplification of zoonotic agents of human diseases. Results of our study in conjunction with abundance in urban/suburban settings, virus isolation from field-collected mosquitoes, and vector competence of Ae. albopictus, highlight the potential involvement of this species in the transmission of a number of arboviruses such as dengue, chikungunya, and Zika to humans. Limited interaction with avian hosts suggests that Ae. albopictus is unlikely to serve as a bridge vector of arboviruses such as West Nile and eastern equine encephalitis in the study region, but that possibility cannot be entirely ruled out.

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.

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.

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.

Passive Tick Surveillance: Exploring Spatiotemporal Associations of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae), Babesia microti (Piroplasmida: Babesiidae), and Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae) Infection in Ixodes scapularis (Acari: Ixodidae).

Ixodes scapularis transmits a group of pathogens, including Borrelia burgdorferi, Babesia microti, and Anaplasma phagocytophilum, the causative agents for Lyme disease, babesiosis, and anaplasmosis, respectively. I. scapularis ticks submitted by state residents to the Connecticut Agricultural Experiment Station-Tick Testing Laboratory between 2015 and 2018 were screened using standard PCR and pathogen-specific primers. Infection and coinfection prevalence in I. scapularis was estimated to assess differences in infection status by life stage (nymph or adult female), county, and year, as well as whether infection with B. burgdorferi changes the likelihood of infection with either B. microti or A. phagocytophilum. Of the 11,254 I. scapularis acquired in Connecticut, 40.7% tested positive for at least one pathogen and the remaining 59.3% were negative. Most I. scapularis ticks tested positive for a single pathogen (33.6%), and only 7.2% were infected with more than one pathogen, of which 93.2% were identified with dual infection and 6.8% tested positive for all three pathogens. Adults were more likely than nymphs to be infected or coinfected with these pathogens. Furthermore, we found that ticks were 74% more likely to be infected with B. microti and 98% more likely to be infected with A. phagocytophilum if infected with B. burgdorferi compared with those not infected. We did not find spatial differences in infection or coinfection prevalence, but between 2015 and 2018, the likelihood that a tick was coinfected increased with time. These results from Connecticut, an endemic state for Lyme disease with long-established populations of I. scapularis, suggest that the increased likelihood of coinfection prevalence over time may have significant implications for clinical diagnosis, course, severity, and treatment of human disease cases.

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.

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.

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.

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.

Integrated control of juvenile Ixodes scapularis parasitizing Peromyscus leucopus in residential settings in Connecticut, United States.

Lyme disease continues to be the most common vector-borne disease in the United States with an estimated 330,000 human cases annually. In the eastern United States, the blacklegged tick, Ixodes scapularis, is the primary vector of the Lyme disease spirochete, Borrelia burgdorferi, and the white-footed mouse, Peromyscus leucopus, is a primary reservoir host. In four residential neighborhoods in Connecticut over three years, we tested the effectiveness of different low-toxicity integrated tick management approaches to control larval and nymphal I. scapularis parasitizing P. leucopus. Combinations of white-tailed deer, Odocoileus virginianus, reduction, broadcast application of the entomopathogenic fungus Metarhizium anisopliae, and distribution of fipronil-based rodent-targeted bait boxes were evaluated against an experimental control. Deer reduction with no other intervention likely forced juvenile I. scapularis to obtain blood meals from available reservoir hosts, resulting in increased exposure of P. leucopus to B. burgdorferi compared to control sites. The M. anisopliae/bait box and the deer reduction/M. anisopliae/bait box treatment combinations resulted in 94% and 85% reductions in larvae parasitizing P. leucopus that tested positive for B. burgdorferi, respectively, compared to control. Deer reduction alone resulted in only a 3% reduction, likely because parasitizing juvenile I. scapularis were not targeted by bait box-delivered fipronil. Unless there is community support to reduce and maintain deer at very low densities (<5 deer/km2), it is clear that a combination of M. anisopliae/fipronil-based bait boxes offers an effective, localized, low-toxicity option for reducing I. scapularis parasitizing P. leucopus without complications from host switching.

A nine-legged tick: Report of a morphological anomaly in the blacklegged tick, Ixodes scapularis (Acari: Ixodidae) from the northeastern United States.

Morphological anomalies have been reported in the blacklegged tick, Ixodes scapularis (Acari: Ixodidae), from the northeastern and upper Midwestern United States, complicating identification of this important vector of human pathogens. Here, we report a case of a local morphological anomaly in I. scapularis, parasitizing a human living in Connecticut. We used a dichotomous morphological key, high-resolution and scanning electron microscopy images, as well as DNA sequencing for identification. The specimen was identified as an adult female I. scapularis with the left leg II split at the trochanter, resulting in two complete legs from the femur to pretarsus giving the appearance of five legs on the left side versus four on the right side. Here we discuss recent reports of morphological anomalies in I. scapularis, and highlight the need for further studies of teratology in this important tick species and its potential implications.