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

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

Jamestown Canyon Virus in Collected Mosquitoes, Maine, United States, 2017-2019.

Jamestown Canyon virus (JCV) is a mosquito-borne arbovirus that circulates in North America. We detected JCV in 4 pools of mosquitoes collected from midcoastal Maine, USA, during 2017-2019. Phylogenetic analysis of a JCV sequence obtained from Aedes cantator mosquitoes clustered within clade A, which also circulates in Connecticut, USA.

Emergence of Ixodes scapularis (Acari: Ixodidae) in a Small Mammal Population in a Coastal Oak-Pine Forest, Maine, USA.

In the United States, surveillance has been key to tracking spatiotemporal emergence of blacklegged ticks [Ixodes scapularis Say (Ixodida:Ixodidae)] and their pathogens such as Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner (Spirochaetales: Spirochaetaceae), the agent of Lyme disease. On the Holt Research Forest in midcoastal Maine, collection of feeding ticks from live-trapped small mammal hosts allowed us to track the emergence and establishment of I. scapularis, 1989-2019. From 1989-1995, we collected only I. angustus Neumann (Ixodida: Ixodidae)(vole tick), Dermacentor variabilis Say (Ixodida: Ixodidae) (American dog tick), and I. marxi Banks (Ixodida: Ixodidae) (squirrel tick) from seven species of small mammals. The most abundant tick host was the white-footed mouse [Peromyscus leucopus Rafinesque (Rodentia:Cricetidae)] followed by the red-backed vole (Myodes gapperi Vigors (Rodentia: Cricetidae)). Emergence of I. scapularis was signaled via the appearance of subadult I. scapularis in 1996. Emergence of B. burgdorferi was signaled through its appearance in I. scapularis feeding on mice in 2005. There was a substantial increase in I. scapularis prevalence (proportion of hosts parasitized) and burdens (ticks/host) on white-footed mice and red-backed voles in 2007. The ~11-yr time-to-establishment for I. scapularis was consistent with that seen in other studies. White-footed mice comprised 65.9% of all captures and hosted 94.1% of the total I. scapularis burden. The white-footed mouse population fluctuated interannually, but did not trend up as did I. scapularis prevalence and burdens. There were concurrent declines in I. angustus and D. variabilis. We discuss these results in the broader context of regional I. scapularis range expansion.

Use of Cervid Serosurveys to Monitor Eastern Equine Encephalitis Virus Activity in Northern New England, United States, 2009-2017.

Vertebrate surveillance for eastern equine encephalitis virus (EEEV) activity usually focuses on three types of vertebrates: horses, passerine birds, and sentinel chicken flocks. However, there is a variety of wild vertebrates that are exposed to EEEV infections and can be used to track EEEV activity. In 2009, we initiated a pilot study in northern New England, United States, to evaluate the effectiveness of using wild cervids (free-ranging white-tailed deer and moose) as spatial sentinels for EEEV activity. In Maine, New Hampshire, and Vermont during 2009-2017, we collected blood samples from hunter-harvested cervids at tagging stations and obtained harvest location information from hunters. U.S. Centers for Disease Control and Prevention processed the samples for EEEV antibodies using plaque reduction neutralization tests (PRNTs). We detected EEEV antibodies in 6 to 17% of cervid samples in the different states and mapped cervid EEEV seropositivity in northern New England. EEEV antibody-positive cervids were the first detections of EEEV activity in the state of Vermont, in northern Maine, and northern New Hampshire. Our key result was the detection of the antibodies in areas far outside the extent of documented wild bird, mosquito, human case, or veterinary case reports of EEEV activity in Maine, New Hampshire, and Vermont. These findings showed that cervid (deer and moose) serosurveys can be used to characterize the geographic extent of EEEV activity, especially in areas with low EEEV activity or with little or no EEEV surveillance. Cervid EEEV serosurveys can be a useful tool for mapping EEEV activity in areas of North America in addition to northern New England.

Support for deer herd reduction on offshore Islands of Maine, U.S.A.

Over the past three decades, citizens of Maine in the northeastern United States have experienced increasing blacklegged tick (Ixodes scapularis) abundance and rising incidence of Lyme and other tick-borne diseases. White-tailed deer (Odocoileus virginianus) overabundance has been considered one cause of the high incidence of tick-borne diseases on offshore islands of New England. Most of Maine's 15 offshore, unbridged island communities have a history of concern about ticks, Lyme disease, and white-tailed deer overabundance, but have been challenged to keep deer numbers down through hunting or culls. This history has led to perennial, often divisive community debates about whether and how to reduce the size of their deer herds. In 2016 we conducted a convenience sample survey of year-round and summer residents of Maine's offshore islands to quantify the level of concern about Lyme disease, and assess the motivations and level of support for deer herd reduction. Among respondents, 84 % agreed Lyme disease was a problem on their island and 61 % supported deer herd reduction. Agreement that Lyme disease was a problem was associated with having acquired tick-borne disease as well as with tick bites without disease. Respondents ranked deer overabundance as a top cause of tick abundance and tick-borne disease and supported deer herd reduction as an approach to reduce the risk of Lyme disease. Other problems associated with deer overabundance (vehicle collisions, damage to landscaping, and damage to forests) also motivated support for deer reduction. Approval of doe permits, an expanded archery season, and sharpshooting as reduction methods was greater than an expanded firearms season. Respondents felt responsibility for tick control fell to the town for the most part, and recognized that multiple factors have contributed to the tick problem in Maine, not just deer.

A Generalized Additive Model Correlating Blacklegged Ticks With White-Tailed Deer Density, Temperature, and Humidity in Maine, USA, 1990-2013.

Geographical range expansions of blacklegged tick [Ixodes scapularis Say (Acari: Ixodidae)] populations over time in the United States have been attributed to a mosaic of factors including 20th century reforestation followed by suburbanization, burgeoning populations of the white-tailed deer [Odocoileus virginianus Zimmerman (Artiodactyla: Cervidae)], and, at the northern edge of I. scapularis' range, climate change. Maine, a high Lyme disease incidence state, has been experiencing warmer and shorter winter seasons, and relatively more so in its northern tier. Maine served as a case study to investigate the interacting impacts of deer and seasonal climatology on the spatial and temporal distribution of I. scapularis. A passive tick surveillance dataset indexed abundance of I. scapularis nymphs for the state, 1990-2013. With Maine's wildlife management districts as the spatial unit, we used a generalized additive model to assess linear and nonlinear relationships between I. scapularis nymph abundance and predictors. Nymph submission rate increased with increasing deer densities up to ~5 deer/km2 (13 deer/mi2), but beyond this threshold did not vary with deer density. This corroborated the idea of a saturating relationship between I. scapularis and deer density. Nymphs also were associated with warmer minimum winter temperatures, earlier degree-day accumulation, and higher relative humidity. However, nymph abundance only increased with warmer winters and degree-day accumulation where deer density exceeded ~2 deer/km2 (~6/mi2). Anticipated increases in I. scapularis in the northern tier could be partially mitigated through deer herd management.

Eastern Equine Encephalitis Virus Seroprevalence in Maine Cervids, 2012-2017.

Eastern equine encephalitis virus (EEEV) first emerged in Maine in the early 2000s and resulted in an epizootic outbreak in 2009. Since 2009, serum samples from cervids throughout Maine have been collected and assessed for the presence of neutralizing antibodies to EEEV to assess EEEV activity throughout the state. We tested 1,119 Odocoileus virginianus (white-tailed deer) and 982 Alces americanus (moose) serum samples collected at tagging stations during the hunting seasons from 2012 to 2017 throughout the state of Maine. Odocoileus virginianus from all 16 counties were EEEV seropositive, whereas A. americanus were seropositive in the northwestern counties of Aroostook, Somerset, Piscataquis, and Franklin counties. Seroprevalence in O. virginianus ranged from 6.6% to 21.2% and in A. americanus from 6.6% to 10.1%. Data from this report in conjunction with findings previously reported from 2009 to 2011 indicate that EEEV is endemic throughout Maine.

Surge in Anaplasmosis Cases in Maine, USA, 2013-2017.

Incidence of human granulocytic anaplasmosis is rising in Maine, USA. This increase may be explained in part by adoption of tick panels as a frequent diagnostic test in persons with febrile illness and in part by range expansion of Ixodes scapularis ticks and zoonotic amplification of Anaplasma phagocytophilum.

Decoupling of Blacklegged Tick Abundance and Lyme Disease Incidence in Southern Maine, USA.

Lyme disease is caused by the bacterial spirochete Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt, and Brenner (Spirocheatales: Spirochaetaceae) which is transmitted through the bite of an infected blacklegged tick Ixodes scapularis Say (Ixodida: Ixodidae). Maine, USA, is a high Lyme disease incidence state, with rising incidence of Lyme disease and other tick-borne illnesses associated with increasing I. scapularis abundance and northward range expansion. Members of the public submitted ticks to a tick identification program (1990-2013). From these passive surveillance data, we characterized temporal trends in I. scapularis submission rate (an index of abundance), comparing Maine's northern tier (seven counties) versus southern tier (nine counties). In the northern tier, the I. scapularis submission rate increased throughout the duration of the time series, suggesting I. scapularis was emergent but not established. By contrast, in the southern tier, submission rate increased initially but leveled off after 10-14 yr, suggesting I. scapularis was established by the mid-2000s. Active (field) surveillance data from a site in the southern tier-bird tick burdens and questing adult tick collections-corroborated this leveling pattern. Lyme disease incidence and I. scapularis submission rate were temporally correlated in the northern but not southern tier. This suggested a decoupling of reported disease incidence and entomological risk.

Prevalence and Genetic Characterization of Deer Tick Virus (Powassan Virus, Lineage II) in Ixodes scapularis Ticks Collected in Maine.

Deer tick virus (DTV) is a genetic variant of Powassan virus (POWV) that circulates in North America in an enzootic cycle involving the blacklegged or "deer tick," Ixodes scapularis, and small rodents such as the white-footed mouse. The number of reported human cases with neuroinvasive disease has increased substantially over the past few years, indicating that POWV may be of increasing public health importance. To this end, we sought to estimate POWV infection rates in questing I. scapularis collected from four health districts in Maine (York, Cumberland, Midcoast, and Central Maine). Infection rates were 1.6%, 1.7%, 0.7%, and 0%, respectively, for adults collected from April to November in 2016. Adults collected in October and November in 2017 from York and Cumberland counties had slightly higher rates of 2.3% and 3.5%, respectively. There was no difference in the number of males verses the number of females infected. All positive samples were of the DTV (lineage II) variant. Phylogenetic analysis was performed on 8 of the 15 DTV sequences obtained in 2016. Deer tick virus from the coastal regions were genetically similar and clustered with virus strains isolated from I. scapularis from New York State and Bridgeport, CT. The two inland viruses were genetically nearly identical and grouped with viruses from Massachusetts, Connecticut, and New York. These results are the first reported infection rates and sequences for POWV in questing ticks collected in Maine and will provide a reference point for future POWV studies.

Seroprevalence of Borrelia burgdorferi, B. miyamotoi, and Powassan Virus in Residents Bitten by Ixodes Ticks, Maine, USA.

We conducted a serosurvey of 230 persons in Maine, USA, who had been bitten by Ixodes scapularis or I. cookei ticks. We documented seropositivity for Borrelia burgdorferi (13.9%) and B. miyamotoi (2.6%), as well as a single equivocal result (0.4%) for Powassan encephalitis virus.

Fatal Deer Tick Virus Infection in Maine.

Deer tick virus (DTV), a genetic variant (lineage II) of Powassan virus, is a rare cause of encephalitis in North America. We report a fatal case of DTV encephalitis following a documented bite from an Ixodes scapularis tick and the erythema migrans rash associated with Lyme disease.

Update on the Distribution of Aedes japonicus in Maine, 2001-15.

The first confirmed collection of Aedes japonicus in Maine was in Cumberland County in 2001. Since that initial collection, it has been found in 7 of Maine's 16 counties between 2001 and 2015. These collections include the northernmost collection of Ae. japonicus in the eastern USA, from Madawaska in Aroostook County. Though mosquito surveillance is limited in the state, it is evident that Ae. japonicus is more widely distributed in Maine than previously believed although its role in arboviral transmission in the region is unknown.

Seasonal Patterns in Eastern Equine Encephalitis Virus Antibody in Songbirds in Southern Maine.

The intent of this study was to assess passerine eastern equine encephalitis virus (EEEv) seroprevalence during the breeding season in southern Maine by testing songbird species identified in the literature as amplifying hosts of this virus. In 2013 and 2014, we collected serum samples from songbirds at a mainland site and an offshore island migratory stopover site, and screened samples for EEEv antibodies using plaque reduction neutralization tests. We compared seasonal changes in EEEv antibody seroprevalence in young (hatched in year of capture) and adult birds at the mainland site, and also compared early season seroprevalence in mainland versus offshore adult birds. EEEv seroprevalence did not differ significantly between years at either site. During the early season (May), EEEv antibody seroprevalence was substantially lower (9.6%) in the island migrant adults than in mainland adults (42.9%), 2013-2014. On the mainland, EEEv antibody seroprevalence in young birds increased from 12.9% in midseason (June-August) to 45.6% in late season (September/October), 2013-2014. Seroprevalence in adult birds did not differ between seasons (48.8% vs. 53.3%). EEEv activity in Maine has increased in the past decade as measured by increased virus detection in mosquitoes and veterinary cases. High EEEv seroprevalence in young birds-as compared to that of young birds in other studies-corresponded with two consecutive active EEEv years in Maine. We suggest that young, locally hatched songbirds be sampled as a part of long-term EEEv surveillance, and provide a list of suggested species to sample, including EEEv "superspreaders."

Human babesiosis, Maine, USA, 1995-2011.

We observed an increase in the ratio of pathogenic Babesia microti to B. odocoilei in adult Ixodes scapularis ticks in Maine. Risk for babesiosis was associated with adult tick abundance, Borrelia burgdorferi infection prevalence, and Lyme disease incidence. Our findings may help track risk and increase the focus on blood supply screening.

Parasitism of mustelids by ixodid ticks (Acari: Ixodidae), Maine and New Hampshire, U.S.A.

Ticks collected from mustelids from four counties in Maine and one in New Hampshire were identified after harvest. Of the 18 fishers Martes pennanti Erxleben, two mink Neovison vison Schreber, and one long-tailed weasel Mustela frenata Lichtenstein, 589 ticks were collected and identified. They were identified as, in order of abundance, Ixodes gregsoni Lindquist, Wu, and Redner (158 larvae, 189 nymphs, four adults), Ixodes cookei Packard (99 larvae, 77 nymphs, six adults), Ixodes scapularis Say (53 adults), Dermacentor variabilis Say (two nymphs), and Ixodes angustus Neumann (one nymph). Seasonally, all but the D. variabilis were collected in winter. This study reports the first record of adult I. scapularis from a M. pennanti in the northeastern United States.

Detection of eastern equine encephalitis virus antibodies in moose (Alces americana), Maine, 2010.

Moose sera were collected from harvested animals during the 2010 hunting season in Maine. Of the 145 serum samples screened by plaque reduction neutralization test (PRNT), 16 (11%) had antibodies to eastern equine encephalitis virus (EEEV). Positive samples were collected from Aroostook County (n=13), Somerset County (n=2), and Piscataquis County (n=1) in northern and central Maine. Preliminary mosquito surveillance revealed the presence of enzootic and bridge vectors mosquitoes, including Culiseta (Climacura) melanura (Coquillett), Aedes (Aedimorphus) vexans (Meigen), and Coquillettidia (Coquillettidia) perturbans (Walker). Select mosquito species were tested by RT-PCR for the presence of EEEV. None were positive. This is the first report of EEEV in moose from Maine.

Effect of a botanical acaricide on Ixodes scapularis (Acari: Ixodidae) and nontarget arthropods.

We tested the effectiveness of the rosemary oil-based insecticide, Eco-Exempt IC2, to control all stages of Ixodes scapularis (Say) in southern Maine. We selected plots in oak-pine forest where I. scapularis is endemic and recorded the abundance of ticks and nontarget arthropods before and after applications of IC2, bifenthrin (a synthetic pyrethroid), and water (reference treatment). Licensed applicators applied high-pressure spray treatments during the summer nymphal and fall adult seasonal peaks. Both acaricides sprayed during the summer nymphal season reduced nymphal I. scapularis/hour to zero. IC2 was as effective as bifenthrin in controlling nymphs through the rest of the nymphal season and also controlled adult ticks 9 mo postspray compared with 16 mo for bifenthrin, and both acaricides reduced larvae through 14 mo postspray. Both acaricides sprayed during the fall adult season reduced adult I. scapularis/hour to zero; IC2 controlled adult ticks 6 mo postspray compared with 1 yr for bifenthrin. Both fall-applied acaricides controlled nymphs 9 mo postspray and reduced larvae up to 10 mo postspray. Impacts on some nontarget arthropods was assessed. Colleoptera, Hymenoptera, and Collembola declined 1 wk postspray in acaricide-treated plots, and in IC2 plots all numbers rebounded by 20 d postspray. For bees and other flower-visiting insects there were no detectable reductions in nests produced, number emerged from nests, or number of foraging visits to flowering plants in IC2 or bifenthrin plots. IC2 was phytotoxic to the leafy portions of select understory plants that appeared to recover by the next growing season.

An epizootic of eastern equine encephalitis virus, Maine, USA in 2009: outbreak description and entomological studies.

From July to September, 2009, an outbreak of eastern equine encephalitis virus (EEEv) occurred in five counties in Maine. The virus was isolated from 15 horses, 1 llama, and pheasants in three separate captive flocks. One wild turkey, screened before translocation, also showed exposure to the virus in January 2010. Two pools of Culiseta melanura (Coquillett) tested positive for EEEv during routine seasonal surveillance in York County in September, but none of the mosquitoes collected during rapid response surveys tested positive. There were more Cs. melanura in July, August, and September 2009 than in preceding (2006-08) and subsequent (2010-11) years. August and September Cs. melanura abundances were correlated with July rainfall, and abundance of all species combined was correlated with total rainfall for the meteorological summer. This outbreak represents a substantial expansion of the range of EEEv activity in northern New England.

Genotypic diversity of an emergent population of Borrelia burgdorferi at a coastal Maine island recently colonized by Ixodes scapularis.

The recent range expansion of Ixodes scapularis has been accompanied by the emergence of Borrelia burgdorferi. The development of genetic diversity in B. burgdorferi at these sites of emergence and its relationship to range expansion is poorly understood. We followed colonization of I. scapularis on a coastal Maine island over a 17-year period. B. burgdorferi's emergence was documented, as was expansion of ospC strain diversity. Ticks collected from rodents and vegetation were examined for the presence of B. burgdorferi. Sequencing and reverse line blot were used to detect B. burgdorferi ospC major groups (oMG). No I. scapularis were found until year four of the study, after which time they increased in abundance. No B. burgdorferi was detected by darkfield microscopy in I. scapularis until 10 years into the study, when 4% of adult ticks were infected. Seven years later, 43% of adult ticks were infected. In 2003, one oMG accounted for 91% of B. burgdorferi strains. This "founder" strain persisted in 2005, but by 2007 was a minority of the 7 oMGs present. Given the island's isolation, gene flow by avian introduction of multiple strains is suggested in the development of B. burgdorferi oMG diversity.