Population dynamics of the Lyme disease bacterium, Borrelia burgdorferi, during rapid range expansion in New York State.

Recent changes in climate and human land-use have resulted in alterations of the geographic range of many species, including human pathogens. Geographic range expansion and population growth of human pathogens increase human disease risk. Relatively little empirical work has investigated the impact of range changes on within-population variability, a contributor to both colonization success and adaptive potential, during the precise time in which populations are colonized. This is likely due to the difficulties of collecting appropriate natural samples during the dynamic phase of migration and colonization. We systematically collected blacklegged ticks (Ixodes scapularis) across New York State (NY), USA, between 2006 and 2019, a time period coinciding with a rapid range expansion of ticks and their associated pathogens including Borrelia burgdorferi, the etiological agent of Lyme disease. These samples provide a unique opportunity to investigate the population dynamics of human pathogens as they expand into novel territory. We observed that founder effects were short-lived, as gene flow from long-established populations brought almost all B. burgdorferi lineages to newly colonized populations within just a few years of colonization. By 7 years post-colonization, B. burgdorferi lineage frequency distributions were indistinguishable from long-established sites, indicating that local B. burgdorferi populations experience similar selective pressures despite geographic separation. The B. burgdorferi lineage dynamics elucidate the processes underlying the range expansion and demonstrate that migration into, and selection within, newly colonized sites operate on different time scales.

Bourbon Virus Transmission, New York, USA.

In July 2019, Bourbon virus RNA was detected in an Amblyomma americanum tick removed from a resident of Long Island, New York, USA. Tick infection and white-tailed deer (Odocoileus virginianus) serosurvey results demonstrate active transmission in New York, especially Suffolk County, emphasizing a need for surveillance anywhere A. americanum ticks are reported.

Borrelia miyamotoi in Human-Biting Ticks, United States, 2013-2019.

During 2013-2019, Borrelia miyamotoi infection was detected in 19 US states. Infection rate was 0.5%-3.2%; of B. miyamotoi-positive ticks, 59.09% had concurrent infections. B. miyamotoi is homogeneous with 1 genotype from Ixodes scapularis ticks in northeastern and midwestern states and 1 from I. pacificus in western states.

Heartland Virus Transmission, Suffolk County, New York, USA.

During 2018, Heartland virus RNA was detected in an Amblyomma americanum tick removed from a resident of Suffolk County, New York, USA. The person showed seroconversion. Tick surveillance and white-tailed deer (Odocoileus virginianus) serosurveys showed widespread distribution in Suffolk County, emphasizing a need for disease surveillance anywhere A. americanum ticks are established or emerging.

Ecology and Epidemiology of Tickborne Pathogens, Washington, USA, 2011-2016.

Tickborne diseases are rare in Washington, USA, and the ecology of these pathogens is poorly understood. We integrated surveillance data from humans and ticks to better describe their epidemiology and ecology. During 2011-2016, a total of 202 tickborne disease cases were reported in Washington residents. Of these, 68 (34%) were autochthonous, including cases of Lyme disease, Rocky Mountain spotted fever, tickborne relapsing fever, and tularemia. During May 2011-December 2016, we collected 977 host-seeking ticks, including Ixodes pacificus, I. angustus, I. spinipalpis, I. auritulus, Dermacentor andersoni, and D. variabilis ticks. The prevalence of Borrelia burgdorferi sensu stricto in I. pacificus ticks was 4.0%; of B. burgdorferi sensu lato, 3.8%; of B. miyamotoi, 4.4%; and of Anaplasma phagocytophilum, 1.9%. We did not detect Rickettsia rickettsii in either Dermacentor species. Case-patient histories and detection of pathogens in field-collected ticks indicate that several tickborne pathogens are endemic to Washington.

Ehrlichia muris in Ixodes cookei Ticks, Northeastern United States, 2016-2017.

Ehrlichia muris is an agent of human ehrlichiosis. To determine its geographic spread in the United States, during 2016-2017, we tested 8,760 ticks from 45 states. A distinct clade of E. muris found in 3 Ixodes cookei ticks from the northeastern United States suggests transmission by these ticks in this region.

Dried whole-plant Artemisia annua slows evolution of malaria drug resistance and overcomes resistance to artemisinin.

Pharmaceutical monotherapies against human malaria have proven effective, although ephemeral, owing to the inevitable evolution of resistant parasites. Resistance to two or more drugs delivered in combination will evolve more slowly; hence combination therapies have become the preferred norm in the fight against malaria. At the forefront of these efforts has been the promotion of Artemisinin Combination Therapy, but despite these efforts, resistance to artemisinin has begun to emerge. In 2012, we demonstrated the efficacy of the whole plant (WP)--not a tea, not an infusion--as a malaria therapy and found it to be more effective than a comparable dose of pure artemisinin in a rodent malaria model. Here we show that WP overcomes existing resistance to pure artemisinin in the rodent malaria Plasmodium yoelii. Moreover, in a long-term artificial selection for resistance in Plasmodium chabaudi, we tested resilience of WP against drug resistance in comparison with pure artemisinin (AN). Stable resistance to WP was achieved three times more slowly than stable resistance to AN. WP treatment proved even more resilient than the double dose of AN. The resilience of WP may be attributable to the evolutionary refinement of the plant's secondary metabolic products into a redundant, multicomponent defense system. Efficacy and resilience of WP treatment against rodent malaria provides compelling reasons to further explore the role of nonpharmaceutical forms of AN to treat human malaria.

A silicone membrane for in vitro feeding of Ixodes scapularis (Ixodida: Ixodidae).

We describe a simple silicone membrane for in vitro feeding of Ixodes scapularis (Say), the main tick vector of Lyme borreliosis in the United States. Membranes are prepared using regenerated cellulose lens paper, which is coated with a controlled layer of silicone by submerging and removing the paper in a defined solution of silicone glue dissolved in toluene. Median engorged weight obtained by I. scapularis fed on defibrinated bovine blood using these membranes was 109 mg (32-198 mg). These ticks laid eggs in the laboratory and larvae hatched 6 wk later. This method simplifies artificial membranes for in vitro feeding of ixodid ticks, which have long required a considerable amount of skill and experience to prepare.

The Spread of Lone Star Ticks (Amblyomma americanum) and Persistence of Blacklegged Ticks (Ixodes scapularis) on a Coastal Island in Massachusetts, USA.

In the northeastern USA, the distribution of lone star ticks (Amblyomma americanum) has expanded northward in recent decades, overlapping with the range of blacklegged ticks (Ixodes scapularis). Blacklegged ticks carry pathogens for diseases such as Lyme, babesiosis, and anaplasmosis, while bites from lone star ticks cause other diseases and the alpha-gal syndrome allergy. Lone star ticks can become so abundant that they are perceived as more of a public health threat than blacklegged ticks. Using the island of Martha's Vineyard, Massachusetts, as a case study, we analyzed data from a total of 1265 yard surveys from 2011 to 2024 to document lone star tick presence and subsequent expansion from two peripheral areas, Chappaquiddick and Aquinnah, to all six towns. The timing of lone star tick expansion on Martha's Vineyard closely matched an increase in tick submissions to a pathogen testing center. At Chappaquiddick, drag sampling carried out in June 2023 and 2024 showed that both tick species were most common at wooded sites, where blacklegged nymphs were somewhat more abundant than lone star nymphs. However, lone star ticks occurred in a wider range of natural and peridomestic habitats than blacklegged nymphs, making them far more challenging for people to avoid and manage.

Effects of Inosine-5'-monophosphate Dehydrogenase (IMPDH/GuaB) Inhibitors on Borrelia burgdorferi Growth in Standard and Modified Culture Conditions.

Borrelia burgdorferi's inosine-5'-monophosphate dehydrogenase (IMPDH, GuaB encoded by the guaB gene) is a potential therapeutic target. GuaB is necessary for B. burgdorferi replication in mammalian hosts but not in standard laboratory culture conditions. Therefore, we cannot test novel GuaB inhibitors against B. burgdorferi without utilizing mammalian infection models. This study aimed to evaluate modifications to a standard growth medium that may mimic mammalian conditions and induce the requirement of GuaB usage for replication. The effects of two GuaB inhibitors (mycophenolic acid, 6-chloropurine riboside at 125 µM and 250 µM) were assessed against B. burgdorferi (guaB+) grown in standard Barbour-Stoenner-Kelly-II (BSK-II) medium (6% rabbit serum) and BSK-II modified to 60% concentration rabbit serum (BSK-II/60% serum). BSK-II directly supplemented with adenine, hypoxanthine, and nicotinamide (75 µM each, BSK-II/AHN) was also considered as a comparison group. In standard BSK-II, neither mycophenolic acid nor 6-chloropurine riboside affected B. burgdorferi growth. Based on an ANOVA, a dose-dependent increase in drug effects was observed in the modified growth conditions (F = 4.471, p = 0.001). Considering higher drug concentrations at exponential growth, mycophenolic acid at 250 µM reduced spirochete replication by 48% in BSK-II/60% serum and by 50% in BSK-II/AHN (p < 0.001 each). 6-chloropurine riboside was more effective in both mediums than mycophenolic acid, reducing replication by 64% in BSK-II/60% serum and 65% in BSK-II/AHN (p < 0.001 each). These results demonstrate that modifying BSK-II medium with physiologically relevant levels of mammalian serum supports replication and induces the effects of GuaB inhibitors. This represents the first use of GuaB inhibitors against Borrelia burgdorferi, building on tests against purified B. burgdorferi GuaB. The strong effects of 6-chloropurine riboside indicate that B. burgdorferi can salvage and phosphorylate these purine derivative analogs. Therefore, this type of molecule may be considered for future drug development. Optimization of this culture system will allow for better assessment of novel Borrelia-specific GuaB inhibitor molecules for Lyme disease interventions. The use of GuaB inhibitors as broadcast sprays or feed baits should also be evaluated to reduce spirochete load in competent reservoir hosts.

Active Surveillance of Powassan Virus in Massachusetts Ixodes scapularis Ticks, Comparing Detection Using a New Triplex Real-Time PCR Assay with a Luminex Vector-Borne Panel.

Powassan virus is an emerging tick-borne pathogen capable of causing severe neuroinvasive disease. As the incidence of human Powassan virus grows both in magnitude and geographical range, the development of sensitive detection methods for diagnostics and surveillance is critical. In this study, a Taqman-based triplex real-time PCR assay was developed for the simultaneous and quantitative detection of Powassan virus and Powassan virus lineage II (deer tick virus) in Ixodes scapularis ticks. An exon-exon junction internal control was built-in to allow for accurate detection of RNA quality and the failure of RNA extraction. The newly developed assay was also applied to survey deer tick virus in tick populations at 13 sites on Cape Cod and Martha's Vineyard Island in Massachusetts. The assay's performance was compared with the Luminex xMAP MultiFLEX Vector-borne Panel 2. The results suggested that the real-time PCR method was more sensitive. Powassan virus infection rates among ticks collected from these highly endemic tick areas ranged from 0.0 to 10.4%, highlighting the fine-scale geographic variations in deer tick virus presence in this region. Looking forward, our PCR assay could be adopted in other Powassan virus surveillance systems.

Detection of heterogeneity of Borrelia burgdorferi in ixodes ticks by culture-dependent and culture-independent methods.

Abilities to detect heterogeneity of ospC genotypes of the Lyme disease spirochete Borrelia burgdorferi in the tick vector by in vitro culture (IVC) and direct PCR (dPCR) were compared. IVC failed to detect one-third of the ospC genotypes detected by dPCR. Among IVC results, common ospC genotypes were overrepresented while occurrence of rare genotypes was underestimated.

What a Tick Can Tell a Doctor: Using the Human-Biting Tick in the Clinical Management of Tick-Borne Disease.

With expanding concern about ticks, there is a general sense of uncertainty about the diagnosis and treatment of tick-borne diseases. The diagnosis process is often based on clinical judgment in conjunction with laboratory testing and can be pathogen specific. Treatments may require disease-dependent approaches, and co-infections complicate or increase the severity of the clinical picture. Measuring exposure indices in the tick has become popular among providers and their patients, though this practice is not universally understood, and certain public health agencies have voiced concerns regarding interpretation and rigor of testing. As many providers subscribe to or recommend these services to aid in pretest risk and exposure assessments, this work sought to clarify the role of pathogen testing human-biting ticks as a complement to the diagnostic pipeline and raises points that must be addressed through future research and interdisciplinary conversation. Future work is needed to develop quality control oversight for tick testing laboratories. Studies on the integration of tick testing with human cases to see how these services affect health outcomes are also needed. Alongside these, improvements in the quality and availability of diagnostics are of critical importance.

Differential Resistance of Borrelia burgdorferi Clones to Human Serum-Mediated Killing Does Not Correspond to Their Predicted Invasiveness.

Reservoir host associations have been observed among and within Borrelia genospecies, and host complement-mediated killing is a major determinant in these interactions. In North America, only a subset of Borrelia burgdorferi lineages cause the majority of disseminated infections in humans. We hypothesize that differential resistance to human complement-mediated killing may be a major phenotypic determinant of whether a lineage can establish systemic infection. As a corollary, we hypothesize that borreliacidal action may differ among human subjects. To test these hypotheses, we isolated primary B. burgdorferi clones from field-collected ticks and determined whether the killing effects of human serum differed among those clones in vitro and/or whether these effects were consistent among human sera. Clones associated with human invasiveness did not show higher survival in human serum compared to noninvasive clones. These results indicate that differential complement-mediated killing of B. burgdorferi lineages is not a determinant of invasiveness in humans. Only one significant difference in the survivorship of individual clones incubated in different human sera was detected, suggesting that complement-mediated killing of B. burgdorferi is usually similar among humans. Mechanisms other than differential human complement-mediated killing of B. burgdorferi lineages likely explain why only certain lineages cause the majority of disseminated human infections.

Passive Surveillance of Human-Biting Ixodes scapularis Ticks in Massachusetts from 2015-2019.

This study aimed to analyze human-biting Ixodes scapularis ticks submitted to TickReport tick testing service from 2015-2019 in Massachusetts to (1) examine possible patterns of pathogen-positive adult and nymphal ticks over time and (2) explore how socioeconomic factors can influence tick submissions. A passive surveillance data set of ticks and tick-borne pathogens was conducted over 5 years (2015-2019) in Massachusetts. The percentages of four tick-borne pathogens: Borrelia burgdorferi, Anaplasma phagocytophilum, Babesia microti, and Borrelia miyamotoi were determined by Massachusetts county and by month and year. Regression models were used to examine the association between zip-code-level socioeconomic factors and submissions. A total of 13,598 I. scapularis ticks were submitted to TickReport from Massachusetts residents. The infection rate of B. burgdorferi, A. phagocytophilum, and B. microti was 39%, 8%, and 7% in adult ticks; 23%, 6%, and 5% in nymphal ticks, respectively. A relatively higher level of education was associated with high tick submission. Passive surveillance of human-biting ticks and associated pathogens is important for monitoring tick-borne diseases, detecting areas with potentially high risks, and providing public information. Socioeconomic factors should be considered to produce more generalizable passive surveillance data and to target potentially underserved areas.

White-Tailed Deer Serum Kills the Lyme Disease Spirochete, Borrelia burgdorferi.

Borrelia burgdorferi is a human pathogen vectored by Ixodes ticks and maintained in nature by a suite of competent vertebrate reservoirs. White-tailed deer (WTD) are considered to be noncompetent reservoirs for B. burgdorferi. Sera from other deer species have been found to be borreliacidal, and similar mechanisms could explain the lack of reservoir competence of WTD. Therefore, we determined whether WTD serum can kill B. burgdorferi. Using an in vitro serum sensitivity assay and subculturing of spirochetes, we demonstrated that WTD serum effectively kills B. burgdorferi. The borreliacidal activity of WTD serum likely contributes to the inability of WTD to efficiently harbor and transmit B. burgdorferi.

Tick Densities and Infection Prevalence on Coastal Islands in Massachusetts, USA: Establishing a Baseline.

Tick-borne diseases and a tick-induced red meat allergy have become increasingly common in the northeastern USA and elsewhere. At the scale of local communities, few studies have documented tick densities or infection levels to characterize current conditions and provide a baseline for further monitoring. Using the town of Nantucket, MA, as a case study, we recorded tick densities by drag sampling along hiking trails in nature preserves on two islands. Nymphal blacklegged ticks (Ixodes scapularis Say) were most abundant at shadier sites and least common in grasslands and scrub oak thickets (Quercus ilicifolia). Lone star ticks (Amblyomma americanum L.) were common on Tuckernuck Island and rare on Nantucket Island, while both tick species were more numerous in 2021 compared to 2020 and 2022. We tested for pathogens in blacklegged nymphs at five sites over two years. In 2020 and 2021, infection levels among the four Nantucket Island sites averaged 10% vs. 19% for Borrelia burgdorferi, 11% vs. 15% for Babesia microti, and 17% (both years) for Anaplasma phagocytophilum, while corresponding levels were significantly greater on Tuckernuck in 2021. Our site-specific, quantitative approach represents a practical example of how potential exposure to tick-borne diseases can be monitored on a local scale.

Lactate Dehydrogenase Inhibitors Suppress Borrelia burgdorferi Growth In Vitro.

Borrelia burgdorferi, the causative agent of Lyme disease, has a highly reduced genome and relies heavily on glycolysis for carbon metabolism. As such, established inhibitors of lactate dehydrogenase (LDH) were evaluated in cultures to determine the extent of their impacts on B. burgdorferi growth. Both racemic and enantiopure (AT-101) gossypol, as well as oxamate, galloflavin, and stiripentol, caused the dose-dependent suppression of B. burgdorferi growth in vitro. Racemic gossypol and AT-101 were shown to fully inhibit spirochetal growth at concentrations of 70.5 and 187.5 µM, respectively. Differences between racemic gossypol and AT-101 efficacy may indicate that the dextrorotatory enantiomer of gossypol is a more effective inhibitor of B. burgdorferi growth than the levorotatory enantiomer. As a whole, LDH inhibition appears to be a promising mechanism for suppressing Borrelia growth, particularly with bulky LDH inhibitors like gossypol.

Human-Biting Ixodes scapularis Submissions to a Crowd-Funded Tick Testing Program Correlate with the Incidence of Rare Tick-Borne Disease: A Seven-Year Retrospective Study of Anaplasmosis and Babesiosis in Massachusetts.

Tick-borne zoonoses pose a serious burden to global public health. To understand the distribution and determinants of these diseases, the many entangled environment-vector-host interactions which influence risk must be considered. Previous studies have evaluated how passive tick testing surveillance measures connect with the incidence of human Lyme disease. The present study sought to extend this to babesiosis and anaplasmosis, two rare tick-borne diseases. Human cases reported to the Massachusetts Department of Health and submissions to TickReport tick testing services between 2015 and 2021 were retrospectively analyzed. Moderate-to-strong town-level correlations using Spearman's Rho (ρ) were established between Ixodes scapularis submissions (total, infected, adult, and nymphal) and human disease. Aggregated ρ values ranged from 0.708 to 0.830 for anaplasmosis and 0.552 to 0.684 for babesiosis. Point observations maintained similar patterns but were slightly weaker, with mild year-to-year variation. The seasonality of tick submissions and demographics of bite victims also correlated well with reported disease. Future studies should assess how this information may best complement human disease reporting and entomological surveys as proxies for Lyme disease incidence in intervention studies, and how it may be used to better understand the dynamics of human-tick encounters.

Detection of Ehrlichia muris eauclairensis in Blacklegged Ticks (Ixodes scapularis) and White-Footed Mice (Peromyscus leucopus) in Massachusetts.

In 2011, Ehrlichia muris eauclairensis (EME) was described as a human pathogen spread by the blacklegged tick, Ixodes scapularis. Until very recently, its reported distribution was limited to the upper midwestern United States, mainly in Minnesota and Wisconsin. In this study, we report the detection of EME DNA in 4 of 16,146 human biting I. scapularis ticks submitted from Massachusetts to a passive tick surveillance program. Active tick surveillance yielded evidence of EME local transmission in the northeastern United States through detection of EME DNA in 2 of 461 host-seeking I. scapularis nymphs, and in 2 white-footed mice (Peromyscus leucopus) of 491 rodent samples collected in the National Ecological Observatory Network (NEON) Harvard Forest site in Massachusetts.