Analysis of Weather Effects on Variability in Lyme Disease Incidence in the Northeastern United States

In this study, variability in reported Lyme disease incidence between 1993 and 2001 was analyzed in seven states in the northeastern part of the USA. Positive significant correlations at p < 0.05 were found in all states between early summer disease incidence and the June moisture index in the region two years prior. These correlations may reflect an enhanced nymphal Ixodes scapularis survival under wetter conditions. In some states, significant correlations were observed related to warmer winter weather a year and a half prior to disease incidence, which may have been due to higher survival and activity levels of the white-footed mouse, an important host for I. scapularis ticks. This revised version was published online in July 2006 with corrections to the Cover Date.     

Abundance and Borrelia burgdorferi-infection Prevalence of Nymphal Ixodes scapularis Ticks along Forest–Field Edges

More than 19,000 human cases of Lyme disease (LD) are reported each year in the United States. Lyme disease cases occur when humans are exposed to the bacterium Borrelia burgdorferi through the bite of an infected ixodid tick. The probability of human exposure to infected ticks results from a combination of human behaviors and entomological risk. Human behaviors include use of tick habitats, use of protective clothing, and grooming for tick removal. Entomological risks include the density of ticks in a habitat and the proportion of these that are infected with B. burgdorferi. Recent studies have suggested that humans are at higher risk of exposure to B. burgdorferi near edges between forests and herbaceous communities, including lawns and old fields, but whether this increased risk is a function of human behaviors, entomological risk, or both, is unknown. We assessed entomological risk across forest–old field edges in Dutchess County, NY. Densities of ticks and of infected ticks were considerably higher within forests than at forest–field edges, and were lowest within fields. Thus, edges between forests and fields do not pose a higher entomological risk than do the forests themselves, although risk at the edge is higher than in herbaceous habitat. Landscapes with abundant edges between forested and herbaceous habitat, and roughly even proportions of both, might attract both heavy human use and pose moderately high entomological risk, and thus could be targeted for mitigation. We suggest that determining appropriate methods for reducing human exposure to LD requires differentiating entomological risk from human behaviors.     

What Ticks Do Under Your Skin: Two-Photon Intravital Imaging of Ixodes Scapularis Feeding in the Presence of the Lyme Disease Spirochete

Lyme disease, due to infection with the Ixodes-tick transmitted spirochete Borrelia burgdorferi, is the most common tick-transmitted disease in the northern hemisphere. Our understanding of the tick-pathogen-vertebrate host interactions that sustain an enzootic cycle for B. burgdorferi is incomplete. In this article, we describe a method for imaging the feeding of Ixodes scapularis nymphs in real-time using two-photon intravital microscopy and show how this technology can be applied to view the response of Lyme borrelia in the skin of an infected host to tick feeding.     

Effect of Climate Change on Lyme Disease Risk in North America

An understanding of the influence of climate change on Ixodes scapularis, the main vector of Lyme disease in North America, is a fundamental component in assessing changes in the spatial distribution of human risk for the disease. We used a climate suitability model of I. scapularis to examine the potential effects of global climate change on future Lyme disease risk in North America. A climate-based logistic model was first used to explain the current distribution of I. scapularis in North America. Climate-change scenarios were then applied to extrapolate the model in time and to forecast vector establishment. The spatially modeled relationship between I. scapularis presence and large-scale environmental data generated the current pattern of I. scapularis across North America with an accuracy of 89% (P < 0.0001). Extrapolation of the model revealed a significant expansion of I. scapularis north into Canada with an increase in suitable habitat of 213% by the 2080s. Climate change will also result in a retraction of the vector from the southern U.S. and movement into the central U.S. This report predicts the effect of climate change on Lyme disease risk and specifically forecasts the emergence of a tickborne infectious disease in Canada. Our modeling approach could thus be used to outline where future control strategies and prevention efforts need to be applied.     

Acquired resistance in dogs to repeated infestation with Ixodes scapularis (Acari: Ixodidae) reduces tick viability and reproductive success

The purpose of this study was to determine whether dogs develop acquired resistance to adult Ixodes scapularis infestation in an experimental model. Five dogs were each infested with ten mating pairs of ticks every week for 7 consecutive weeks, another five dogs were each infested with ten mating pairs once every 2 weeks for 10 weeks and four dogs served as controls not exposed to ticks. All ticks were allowed to feed to repletion and were collected only after dropping from the host. Several variables were measured to determine the extent of blood feeding success. Regression analysis indicated that the engorgement success, survival and mean tick engorgement weight declined with repeated infestation in both groups of dogs (p<0.05). Tick oviposition as well as the F1 viability declined with each successive infestation in both groups. These results suggest that repeated infestation with I. scapularis elicits a protective immune response against tick feeding and could serve as a limiting factor in the spread and transmission of Borrelia burgdorferi.     

Field and laboratory studies on the timing of oviposition and hatching of the western black-legged tick, Ixodes pacificus (Acari: Ixodidae)

The timing of oviposition and hatching of Ixodes pacificus was investigated in the field and at constant temperatures in the laboratory. Replete females held at temperatures between 9 and 29°C began depositing eggs a mean of 9–70 days after drop off. Egg masses held between 12 and 25°C commenced hatching 25–178 days after the onset of oviposition. Eggs held at 9 or 29°C did not hatch. The lower temperature thresholds for development (LTD) for oviposition and hatching were 6.5 and 9°C, respectively. The number of degree days required for oviposition and hatching was 173 and 588, respectively. Replete females placed in the field on 2 December through to 8 March deposited eggs from 2 February through to 24 April; the eggs commenced hatching between 2 July and 21 August. Unfed larvae from two of 20 egg masses survived through the winter and fed readily when exposed to deer mice (Peromyscus maniculatus) on 22 April. Replete larvae were returned to the field and moulted between 9 and 21 August. Larvae exposed to deer mice in August, 4 weeks after hatching, also fed readily. Although further studies are needed to clarify the timing of nymphal development, the present study suggests that I. pacificus requires more than 1 year to complete its life cycle.     

Pathogenicity of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> (Deuteromycetes) and permethrin to <Emphasis Type="Italic">Ixodes scapularis</Emphasis> (Acari: Ixodidae) nymphs

Effectiveness of the entomopathogenic fungus Metarhizium anisopliae, for controlling nymphal Ixodes scapularis, was tested in laboratory and field trials. In the laboratory, M. anisopliae (Metschnikoff) Sorokin strain ESC1 was moderately pathogenic, with an LC₅₀ of 10⁷ spores/ml and induced 70% mortality at 10⁹ spores/ml. In a field study, however, 10⁹ spores/ml M. anisopliae did not effectively control questing I. scapularis nymphs, and significant differences were not detected in pre- and post-treatment densities. For nymphs collected and returned to the laboratory for observation, mortality was low in treatment groups, ranging from 20 to 36%. To assess whether a chemical acaricide would synergistically enhance pathogenicity of the fungus, we challenged unfed nymphal I. scapularis with combinations of M. anisopliae and permethrin, a relatively safe pyrethroid acaricide, in two separate bioassays. Significant interactions between M. anisopliae and permethrin were not observed, supporting neither synergism nor antagonism.     

Temporal Distribution of the Annual Nymphal Stock of Ixodes Ricinus Ticks

The human risk of contracting Lyme disease or other tick borne diseases transmitted by the tick species Ixodes ricinus is broadly linked to the tick nymph density. The study was performed in Rambouillet forest (Yvelines, France), a known focus of Lyme borreliosis, from January 1997 to December 1999. We used a nymph sampling methodology which permitted us to obtain a monthly nymph density index (from 0 to 5). Studying the seasonal nymph and larval activity patterns and estimating the larval developmental duration, we demonstrate the existence of an annual nymphal stock. Secondly, we elucidate how this stock is distributed throughout the year, month by month. Its distribution is principally dependent on two factors: the monthly mean ambient temperature and the proportion of active nymphs which find a host each month. Expected monthly nymph densities derived from a theoretical model describing the temperature-dependent stock distribution gave a good fit to the observed densities, accounting for between 76–86% of the monthly variation in observed nymph densities. Predicting the temporal distribution of nymph activity within a stable Lyme borreliosis focus enables more precise identification of risk periods.     

Accelerated phenology of blacklegged ticks under climate warming

The phenology of tick emergence has important implications for the transmission of tick-borne pathogens. A long lag between the emergence of tick nymphs in spring and larvae in summer should increase transmission of persistent pathogens by allowing infected nymphs to inoculate the population of naive hosts that can subsequently transmit the pathogen to larvae to complete the transmission cycle. In contrast, greater synchrony between nymphs and larvae should facilitate transmission of pathogens that do not produce long-lasting infections in hosts. Here, we use 19 years of data on blacklegged ticks attached to small-mammal hosts to quantify the relationship between climate warming and tick phenology. Warmer years through May and August were associated with a nearly three-week advance in the phenology of nymphal and larval ticks relative to colder years, with little evidence of increased synchrony. Warmer Octobers were associated with fewer larvae feeding concurrently with nymphs during the following spring. Projected warming by the 2050s is expected to advance the timing of average nymph and larva activity by 8–11 and 10–14 days, respectively. If these trends continue, climate warming should maintain or increase transmission of persistent pathogens, while it might inhibit pathogens that do not produce long-lasting infections.     

Densities of Ixodes ricinus ticks (Acari: Ixodidae) on moorland vegetation communities in the UK

Unfed (questing) Ixodes ricinus ticks were collected by blanket dragging on a monthly basis from heather-dominated, Vaccinium-dominated and bracken-dominated vegetation communities from two different biogeographical regions of the UK (the Quantock Hills in Somerset, south west England and the North York Moors, north east England) throughout the spring and summer months of 1991 and 1992. Eighteen sites were monitored across the two regions and a total of 1920 blanket drags were carried out. Vaccinium sites showed high tick densities at all life stages, as did bracken sites. Significantly lower numbers of larval and nymphal ticks per drag were collected on heather sites than were collected on either Vaccinium (bilberry/whortleberry) or bracken sites, while similar numbers of adult ticks per drag were collected from each of the three vegetation communities. There was no significant difference between the mean numbers of any tick life stage collected on the Quantock Hills and those collected on the North York Moors on these vegetation communities or between the mean numbers of any tick life stage collected in 1991 and those collected in 1992 on these vegetation communities.     

Molecular and pharmacological characterization of two D(1)-like dopamine receptors in the Lyme disease vector, Ixodes scapularis

Advancements in tick neurobiology may impact the development of acaricides to control those species that transmit human and animal diseases. Here, we report the first cloning and pharmacological characterization of two neurotransmitter binding G protein-coupled receptors in the Lyme disease (blacklegged) tick, Ixodes scapularis. The genes IscaGPRdop1 and IscaGPRdop2 were identified in the I. scapularis genome assembly and predicted as orthologs of previously characterized D₁-like dopamine receptors in the fruit fly Drosophila melanogaster and honeybee Apis mellifera. Heterologous expression in HEK 293 cells demonstrated that each receptor functioned as a D₁-like dopamine receptor because significant increases in levels of intracellular cyclic adenosine monophosphate (cAMP) were detected following dopamine treatment. Importantly, the receptors were distinct in their pharmacological properties regarding concentration-dependent response to dopamine, constitutive activity, and response to other biogenic amines. Exposure to a variety of dopamine receptor agonists and antagonists further demonstrated a D₁-like pharmacology of these dopamine receptors and highlighted their differential activities in vitro.     

A Preliminary Linkage Map of the Tick, Ixodes scapularis

A linkage map of the Ixodes scapularis genome was constructed based upon segregation amongst 127 loci. These included 84 random amplified polymorphic DNA (RAPD) markers, 32 Sequence-Tagged RAPD (STAR) markers, 5 cDNAs, and 5 microsatellites in 232 F₁ intercross progeny from a single, field-collected P₁ female. A preliminary linkage map of 616 cM was generated across 14 linkage groups with one marker every 10.8 cM. Assuming a genome size of ∼10⁹ bp, the relationship of physical to genetic distance is ∼300 kb/cM in the I. scapularis genome. This revised version was published online in July 2006 with corrections to the Cover Date.     

Close to home: a history of Yale and Lyme disease

Yale scientists played a pivotal role in the discovery of Lyme disease and are credited as the first to recognize, name, characterize, and treat the affliction. Today, Lyme disease is the most commonly reported vector-borne illness in the United States, affecting approximately 20,000 people each year, with the incidence having doubled in the past 10 years [1]. Lyme disease is the result of a bacterial infection transmitted to humans through the bite of an infected deer tick, which typically results in a skin rash at the site of attack. While most cases, when caught early, are easily treated by antibiotic therapy, delayed treatment can lead to serious systemic side effects involving the joints, heart, and central nervous system. Here we review Yale's role in the discovery and initial characterization of Lyme disease and how those early discoveries are crucial to our current understanding of the disease.     

Use of Refrigeration as a Practical means to Preserve Viability of in Vitro-Cultured IDE8 Tick Cells

In vitro cultivation of the IDE8 cell line, derived from embryonic Ixodes scapularis ticks, constitutes an important system for the study of tick-borne pathogens, as these cells support growth of rickettsial species which are not normally transmitted by this tick. However, since cryopreservation of IDE8 cells is not always successful, there is a need to develop alternative ways to preserve these cells. In the present study, a suspension of IDE8 cells in culture medium was kept under refrigeration at 4°C for up to 60 days. Every 15 days, the suspension was mixed and aliquots were re-cultured in 2-ml tubes, under standardized conditions. In addition, three techniques for cryopreservation, using two different cryoprotectants (DMSO and glycerol), were evaluated. Medium changes were carried out every week and subculturing every 2 weeks. The development of cultures and their respective subcultures, after returning to standard culture temperature, was evaluated by percentage viability and by cellular morphology evaluated in Giemsa-stained cytocentrifuge smears. All cultures and subcultures appeared healthy, showing growth rates comparable to cultures that had not been kept under refrigeration. The results demonstrated that storage under refrigeration at 4°C is an efficient method for preservation of IDE8 cells for up to 60 days and that refrigeration may be preferable to cryopreservation for short-term preservation of IDE8 cells.     

The influence of landscape diversity and heterogeneity on spatial dynamics of agrobiont linyphiid spiders: An individual-based model

Spiders are important generalist predators in natural pest control. However, agricultural fields are highly disturbed and ephemeral habitats, which present a number of challenges to the organisms living there; likewise landscape diversity and heterogeneity are also thought to be important factors in determining spider spatial dynamics. To investigate the interactions between these factors, we present an individual-based simulation model, which integrates life history characteristics of a typical agrobiont linyphiid spider with a dynamic spatially explicit landscape representation. The landscape contains several habitat types of varying quality and varies in time and space. Simulations showed that spatial landscape diversity (number of habitat types available for the spiders) is crucial for the persistence of spiders, but that spatial heterogeneity (spatial arrangement of patches) only had little impact on spider abundance. The necessary landscape diversity could either be provided by a diverse crop rotation or by including refuges in the form of less frequently managed habitats in the landscape. The presence of refuges greatly boosted numbers of spiders in the landscape as a whole. The most important characteristics of refuge were sanctuary from pesticides and extra prey availability, whereas tillage frequency mattered less. The simulations indicated that agrobiont linyphiids combination of high dispersal abilities and high reproductive rate enables it to exploit the transient resources of the different habitats in the agricultural landscape.     

A Basis for Spatial and Social Patterns in Ant Species: Dynamics and Mechanisms of Aggregation

Aggregation is usually studied in functional terms, forgetting mechanisms. In this paper, experimental results on the ant Lasius niger, complemented by a model, allow us to understand the mechanisms responsible for aggregation and to study the influence of the population density on this phenomenon. The results show (1) a high level of aggregation and the emergence of a large cluster; (2) that aggregation results from an amplification mechanism—the greater the number of ants inside a cluster, the greater the time spent by one ant in this cluster; and (3) that population density has only a weak influence on the aggregation process. This method of analysis and these results can certainly be extended not only in social insects but also in other species, like subsocial arthropods.     

Detection of Lyme disease spirochete,Borrelia burgdorferi sensu lato,including three novel genotypes in ticks (Acari: Ixodidae) collected from songbirds (Passeriformes) across Canada

Lyme disease is reported across Canada, but pinpointing the source of infection has been problematic. In this three‐year, bird‐tick‐pathogen study (2004–2006), 366 ticks representing 12 species were collected from 151 songbirds (31 passerine species/subspecies) at 16 locations Canada‐wide. Of the 167 ticks/pools tested, 19 (11.4%) were infected with Borrelia burgdorferi sensu lato (s.l.). Sequencing of the rrf‐rrl intergenic spacer gene revealed four Borrelia genotypes: B. burgdorferi sensu stricto (s.s.) and three novel genotypes (BC genotype 1, BC genotype 2, BC genotype 3). All four genotypes were detected in spirochete‐infected Ixodes auritulus (females, nymphs, larvae) suggesting this tick species is a vector for B. burgdorferi s.l. We provide first‐time records for: ticks in the Yukon (north of 60° latitude), northernmost collection of Amblyomma americanum in North America, and Amblyomma imitator in Canada. First reports of bird‐derived ticks infected with B. burgdorferi s.l. include: live culture of spirochetes from Ixodes pacificus (nymph) plus detection in I. auritulus nymphs, Ixodes scapularis in New Brunswick, and an I. scapularis larva in Canada. We provide the first account of B. burgdorferi s. l. in an Ixodes muris tick collected from a songbird anywhere. Congruent with previous data for the American Robin, we suggest that the Common Yellowthroat, Golden‐crowned Sparrow, Song Sparrow, and Swainson's Thrush are reservoir‐competent hosts. Song Sparrows, the predominant hosts, were parasitized by I. auritulus harboring all four Borrelia genotypes. Our results show that songbirds import B. burgdorferi s.l.‐infected ticks into Canada. Bird‐feeding I. scapularis subadults were infected with Lyme spirochetes during both spring and fall migration in eastern Canada. Because songbirds disperse millions of infected ticks across Canada, people and domestic animals contract Lyme disease outside of the known and expected range.     

Problems of isolating Borrelia burgdorferi from ticks collected in United Kingdom foci of Lyme disease

Abstract. Many isolates of Borrelia burgdorferi have been obtained from ticks and vertebrate tissues collected in North America and continental Europe but only one established culture of United Kingdom Borrelia burgdorferi has been recorded. In this paper we report the isolation of B.burgdorferi from one of 108 tick pools representing 733 ticks and eighty-four tissue samples from twenty-six rodents collected in the U.K., and the subsequent failure to establish the isolate (from ticks collected in Fordingbridge) in culture. In contrast, using identical techniques and culture medium, B.burgdorferi was isolated from one of seven tick pools collected in Switzerland, and from a single pool of ticks collected in Slovakia, and both isolates were successfully passaged. Analysis of questing I.ricinus collected from Fordingbridge by direct immunofluorescence showed 6/32 (19%) of adults and 8/108 (7%) of nymphs were positive for B. burgdorferi , although only one nymph contained ≥ 1000 spirochaetes. To examine further the problem of isolating U.K. B.burgdorferi , twelve Ixodes ricinus tick samples from Fordingbridge, a recognized focus of Lyme disease, were subjected to isolation and culturing techniques, and the procedures monitored by use of the polymerase chain reaction (PCR). Whereas 11/12 samples were PCR positive after 2 weeks in culture, only one was PCR positive after 4 weeks. Motile spirochaetes were not visible by dark-field microscopy in any of the cultures. The results indicate that the standard BSK II medium routinely used to isolate and culture B. burgdorferi does not readily support the replication of the Borrelia species endemic to the U.K.     

Mating,male Ixodes scapularis express several genes including those with sequence similarity to immunoglobulin-binding proteins and metalloproteases

We created a cDNA library from mating, male Ixodes scapularis ticks and screened the library with a subtracted probe to eliminate genes common to feeding female and mating male I. scapularis ticks. A total of seven unique cDNAs were identified in this screen. One cDNA had sequence similarity to the IGBP-MC gene from Rhipicephalus appendiculatus, and another cDNA potentially encodes a protein with similarity to metalloproteases. RT-PCR, using RNA isolated from male and female I. scapularis ticks, confirmed that these genes are expressed in male, but not female ticks. The remaining five cDNAs did not match any sequences in the GenBank database. This revised version was published online in July 2006 with corrections to the Cover Date.     

Climate change and the potential for range expansion of the Lyme disease vector Ixodes scapularis in Canada

We used an Ixodes scapularis population model to investigate potential northward spread of the tick associated with climate change. Annual degree-days >0 degrees C limits for I. scapularis establishment, obtained from tick population model simulations, were mapped using temperatures projected for the 2020s, 2050s and 2080s by two Global Climate Models (the Canadian CGCM2 and the UK HadCM3) for two greenhouse gas emission scenario enforcings 'A2'and 'B2' of the Intergovernmental Panel on Climate Change. Under scenario 'A2' using either climate model, the theoretical range for I. scapularis establishment moved northwards by approximately 200 km by the 2020s and 1000 km by the 2080s. Reductions in emissions (scenario 'B2') had little effect on projected range expansion up to the 2050s, but the range expansion projected to occur between the 2050s and 2080s was less than that under scenario 'A2'. When the tick population model was driven by projected annual temperature cycles (obtained using CGCM2 under scenario 'A2'), tick abundance almost doubled by the 2020s at the current northern limit of I. scapularis, suggesting that the threshold numbers of immigrating ticks needed to establish new populations will fall during the coming decades. The projected degrees of theoretical range expansion and increased tick survival by the 2020s, suggest that actual range expansion of I. scapularis may be detectable within the next two decades. Seasonal tick activity under climate change scenarios was consistent with maintenance of endemic cycles of the Lyme disease agent in newly established tick populations. The geographic range of I. scapularis-borne zoonoses may, therefore, expand significantly northwards as a consequence of climate change this century.