Salinity stress increases the severity of ranavirus epidemics in amphibian populations.

The stress-induced susceptibility hypothesis, which predicts chronic stress weakens immune defences, was proposed to explain increasing infectious disease-related mass mortality and population declines. Previous work characterized wetland salinization as a chronic stressor to larval amphibian populations. Thus, we combined field observations with experimental exposures quantifying epidemiological parameters to test the role of salinity stress in the occurrence of ranavirus-associated mass mortality events. Despite ubiquitous pathogen presence (94%), populations exposed to salt runoff had slightly more frequent ranavirus related mass mortality events, more lethal infections, and 117-times greater pathogen environmental DNA. Experimental exposure to chronic elevated salinity (0.8-1.6 g l-1 Cl-) reduced tolerance to infection, causing greater mortality at lower doses. We found a strong negative relationship between splenocyte proliferation and corticosterone in ranavirus-infected larvae at a moderate elevation of salinity, supporting glucocorticoid-medicated immunosuppression, but not at high salinity. Salinity alone reduced proliferation further at similar corticosterone levels and infection intensities. Finally, larvae raised in elevated salinity had 10 times more intense infections and shed five times as much virus with similar viral decay rates, suggesting increased transmission. Our findings illustrate how a small change in habitat quality leads to more lethal infections and potentially greater transmission efficiency, increasing the severity of ranavirus epidemics.

Ranavirus infection dynamics and shedding in American bullfrogs: consequences for spread and detection in trade.

American bullfrogs Lithobates catesbeianus are thought to be important in the global spread of ranaviruses-often lethal viruses of cold-blooded vertebrates-because they are commonly farmed, dominate international trade, and may be 'carriers' of ranavirus infections. However, whether American bullfrogs are easily infected and maintain long-lasting ranavirus infections, or are refractory to or rapidly clear infections, remains unknown. We tracked the dynamics of ranavirus in American bullfrogs through time and with temperature in multiple types of samples and also screened shipments from commercial suppliers to determine whether we could detect subclinical infections. Collectively, we found that tadpoles and juveniles were commonly infected at moderate doses, and while some died, others controlled and appeared to clear their infections. Some individuals, however, harbored subclinical infections for up to 49 d, suggesting that American bullfrogs may be important carriers. Indeed, tadpoles and metamorphosed frogs from 2 of 5 commercial suppliers harbored subclinicial infections. Juveniles at warmer temperatures had less intense but still persistent infections. Because diagnostic performance was strongly related to infection intensity, non-lethal samples (i.e. tail or toe clips, swabs, and environmental DNA) had only a moderate chance of detecting subclinical infections. Even internal tissues may fail to detect subclinical infections. However, viral shedding was correlated with the intensity of infection, so while subclinically infected tadpoles shed virus for 35-49 d, the low levels might lead to little transmission. We suggest that a quantitative focus on virus dynamics within hosts can provide a more nuanced view of ranavirus infections and the risk presented by American bullfrogs in trade.

Heterogeneities in the infection process drive ranavirus transmission.

Transmission is central to our understanding and efforts to control the spread of infectious diseases. Because transmission generally requires close contact, host movements and behaviors can shape transmission dynamics: random and complete mixing leads to the classic density-dependent model, but if hosts primarily interact locally (e.g., aggregate) or within groups, transmission may saturate. Manipulating host behavior may thus change both the rate and functional form of transmission. We used the ranavirus-wood frog (Lithobates sylvaticus) tadpole system to test whether transmission rates reflect contacts, and whether the functional form of transmission can be influenced by the distribution of food in mesocosms (widely dispersed, promoting random movement and mixing vs. a central pile, promoting aggregations). Contact rates increased with density, as expected, but transmission rapidly saturated. Observed rates of transmission were not explained by observed contact rates or the density-dependent model, but instead transmission in both treatments followed models allowing for heterogeneities in the transmission process. We argue that contacts were not generally limiting, but instead that our results are better explained by heterogeneities in host susceptibility. Moreover, manipulating host behavior to manage the spread of infectious disease may prove difficult to implement.

Detection of the emerging amphibian pathogens Batrachochytrium dendrobatidis and ranavirus in Russia.

In a population of the European common toad Bufo bufo from a rural pond in the region of Lake Glubokoe Regional Reserve in Moscow province, Russia, unexplained mass mortality events involving larvae and metamorphs have been observed over a monitoring period of >20 yr. We tested toads from this and a nearby site for the emerging amphibian pathogens Batrachochytrium dendrobatidis (Bd) and ranavirus (Rv). Both pathogens were detected, and at the rural pond site, with the above-noted losses and decline in toad breeding success, 40% of B. bufo metamorphs were Bd positive, 46% were Rv positive and 20% were co-infected with both pathogens. Toad metamorphs from a neighbouring water body were also Bd and Rv positive (25 and 55%, respectively). This is the first confirmation of these pathogens in Russia. Questions remain as to the origins of these pathogens in Russia and their roles in documented mass mortality events.

Responses of juvenile blacklegged ticks (Acari: Ixodidae) to hosts of varying quality.

Blacklegged ticks (Ixodes scapularis) are the most medically and economically important vectors in North America. Each of their 3 life stages requires a blood meal from one of many potential host species, during which they can acquire or transmit pathogens. Host species, however, vary tremendously in their quality for ticks, as measured by differences in feeding and molting success. There should be clear fitness benefits for ticks that preferentially feed upon high-quality hosts (e.g., white-footed mice, Peromyscus leucopus), or at least avoid feeding on very low-quality hosts (e.g., Virginia opossums, Didelphis virginiana). Indeed, laboratory experiments have found some evidence of host preferences in I. scapularis; but these involve presenting ticks with hosts simultaneously and measuring movement towards hosts on a horizontal plane. In nature, however, host-seeking ticks encounter hosts sequentially and their movements are principally in a vertical plane. Here, we present the results of a study in which we measured the vertical movements of host-seeking juvenile blacklegged ticks before and after a host (P. leucopus, Tamias striatus, Sciurus carolinensis, or D. virginiana) was present, and whether the strength of their responses varies with host quality. We found ticks did not measurably alter the speed of their vertical movement in the presence of any hosts, regardless of host quality. Both larvae and nymphs quested slightly higher in the presence of hosts, but this did not vary by host species. These results call into question the existence of active host preferences, at least in this stage of the host-seeking process.

Environmental DNA concentrations vary greatly across productive and degradative conditions, with implications for the precision of population estimates.

Population size is an important metric to inform the conservation and management of species. For aquatic species, environmental DNA (eDNA) concentration has been suggested for non-invasively estimating population size. However, many biotic and abiotic factors simultaneously influence the production and degradation of eDNA which can alter the relationship between population size and eDNA concentration. We investigated the influence of temperature, salinity, and ranavirus infection on eDNA concentrations using tadpole mesocosms. Using linear regression models, we tested the influence of each experimental treatment on eDNA concentrations at three time points before and during epidemics. Prior to infection, elevated temperatures lowered eDNA concentrations, indicating that degradation was the driving force influencing eDNA concentrations. During early epidemics, no treatments strongly influenced eDNA concentrations and in late epidemics, productive forces dominated as ranavirus intensity and dead organisms increased eDNA concentrations. Finally, population size was only an important predictor of eDNA concentration in late epidemics and we observed high levels of variation between samples of replicate mesocosms. We demonstrate the complexities of several interacting factors influencing productive and degradative forces, variation in influences on eDNA concentration over short time spans, and examine the limitations of estimating population sizes from eDNA with precision in semi-natural conditions.

Attitudes and Behavioral Intentions of Pet Amphibian Owners About Biosecurity Practices.

Global trade has been linked with the emergence of novel pathogens and declines in amphibian populations worldwide. The potential for pathogen transmission within and between collections of captive amphibians and spillover to wild populations makes it important to understand the motivations, knowledge, attitudes and behaviors of pet amphibian owners. We surveyed US pet amphibian owners to understand their characteristics and evaluated whether and how they were associated with behavioral intentions to adopt biosecurity practices. We found that the majority of pet amphibian owners are aware of the threat of emerging pathogens, concerned about potential spillover of pathogens from captive to wild populations and willing to adopt biosecurity practices to mitigate pathogen threats. Intentions to adopt such practices were driven more by psychosocial constructs such as attitudes, perceptions and beliefs than demographic characteristics. Pet amphibian owners also expressed a strong interest in acquiring, and willingness to pay a price premium for, certified disease-free animals. These findings advance our understanding of the characteristics, motivations and behaviors of pet owners, a key stakeholder in global amphibian trade, which could help to inform new policies and outreach strategies to engage them in mitigating pathogen threats. Moreover, our results imply the economic viability of a market-based program to promote pathogen-free, sustainable trade of amphibians.

Overwintering survival of nymphal Ixodes scapularis (Acari: Ixodidae) under natural conditions.

Blacklegged ticks (Ixodes scapularis Say) are exquisitely sensitive to very cold and dry conditions. For this reason it has long been assumed that climatic differences among locations and within microhabitats have a strong influence on variation in their survival over winter. This assumption, however, rests largely on laboratory exposures and on broad-scale associations between climatic variables and the observed distributions of ticks. We present the results of a study of the overwintering survival of I. scapularis nymphs in their natural environment from October through May in two locations in New York State using a repeated sampling strategy to determine when mortality occurred, and whether those events coincide with extreme conditions. We then fit these data to a simple, flexible statistical model in which the hazard of mortality varies with measurable conditions, here minimum daily temperature and mean daily relative humidity. Regardless of winter conditions, > 80% of ticks survived at both sites. A model with constant hazard (i.e., independent of temperature and humidity) was best supported by the data. Although models with hazard increasing at temperatures below 0 deg C and at > 90% relative humidity provided slightly better fits to the data, these models were less parsimonious. These results weaken the expectation that cold-related overwintering mortality necessarily plays a major role in restricting populations of these ticks and thus, risk of tick-borne zoonoses.

Molting success of Ixodes scapularis varies among individual blood meal hosts and species.

The blacklegged tick (Ixodes scapularis) is an important vector of emerging human pathogens. It has three blood-feeding stages, as follows: larva, nymph, and adult. Owing to inefficient transovarial transmission, at least for the Lyme disease agent (Borrelia burgdorferi), larval ticks rarely hatch infected, but they can acquire infection during their larval blood meal. Nymphal ticks are primarily responsible for transmitting pathogens to hosts, including humans. The transition from uninfected host-seeking larva to infectious host-seeking nymph is therefore a key aspect of human risk of infection. It can be divided into a series of steps, as follows: finding a host, taking a blood meal, becoming infected, molting, and overwintering. The chance of succeeding in each of these steps may depend on the species identity of the blood meal host. We used a Bayesian method to estimate the molting success of larval I. scapularis collected from four commonly parasitized species of birds and eight commonly parasitized small and mid-sized mammals found in the forests of Dutchess County, New York. We show that molting success varies substantially among host species; white-footed mice, veeries, and gray catbirds support particularly high molting success, whereas ticks feeding on short-tailed shrews, robins, and wood thrushes were less successful. We also show that larval molting success varies substantially between individual blood meal hosts, and that this intraspecific variability is much higher in some species than in others. The causes of both inter- and intraspecific variation in molting success remain to be determined.

Pooled samples and eDNA-based detection can facilitate the "clean trade" of aquatic animals.

The regional and international trade of live animals facilitates the movement, spillover, and emergence of zoonotic and epizootic pathogens around the world. Detecting pathogens in trade is critical for preventing their continued movement and introduction, but screening a sufficient fraction to ensure rare infections are detected is simply infeasible for many taxa and settings because of the vast numbers of animals involved-hundreds of millions of live animals are imported into the U.S.A. alone every year. Batch processing pools of individual samples or using environmental DNA (eDNA)-the genetic material shed into an organism's environment-collected from whole consignments of animals may substantially reduce the time and cost associated with pathogen surveillance. Both approaches, however, lack a framework with which to determine sampling requirements and interpret results. Here I present formulae for pooled individual samples (e.g,. swabs) and eDNA samples collected from finite populations and discuss key assumptions and considerations for their use with a focus on detecting Batrachochytrium salamandrivorans, an emerging pathogen that threatens global salamander diversity. While empirical validation is key, these formulae illustrate the potential for eDNA-based detection in particular to reduce sample sizes and help bring clean trade into reach for a greater number of taxa, places, and contexts.

Environmental Drivers of Questing Activity of Juvenile Black-Legged Ticks (Acari: Ixodidae): Temperature, Desiccation Risk, and Diel Cycles.

Vector feeding behavior can have a profound influence on the transmission of vector-borne diseases. In the case of black-legged ticks, Ixodes scapularis Say, which vectors the agents of Lyme disease, babesiosis, and other pathogens, the timing and propensity of questing can determine which hosts are fed upon as well as the risk of contact with humans. Yet we know little about the controls and constraints on tick host-finding behavior under natural conditions. Ticks must balance the need to quest for blood meal hosts with the risk of desiccation, all on a fixed energy budget. Prior research, primarily in the laboratory, has shown that questing activity varies with conditions (e.g., temperature, relative humidity), light-dark cycles, and energy reserves, but the findings have been idiosyncratic and the dominant factor(s) in nature remains unknown. We measured questing activity of nymphs and larvae throughout the day and night and over several weeks in enclosures across a range of suitable tick habitats within a site in the Northeast. Activity of nymphs increased slightly during dawn and dusk, opposite of larvae, and declined slightly with air temperature and rain, but these patterns were weak and inconsistent among replicate sites. Rather it appears a fraction of ticks were questing most of the time, regardless of conditions. Our study suggests neither climatic conditions or light-dark cycles have appreciable influence on tick questing behavior.

Spatial and temporal patterns of Ambystoma tigrinum virus (ATV) prevalence in tiger salamanders Ambystoma tigrinum nebulosum.

Amphibian ranaviruses have been documented as causes of mass mortality in amphibian populations throughout the world. The temporal and spatial dynamics of ranavirus infections when epidemics are not apparent remains unclear. To address this question, we collected tissue samples from 2003 to 2006 in 4 geographically separated tiger salamander Ambystoma tigrinum nebulosum host populations on the Kaibab Plateau in northern Arizona. We tested for Ambystoma tigrinum virus (ATV), a lethal ranavirus of tiger salamanders, calculated ATV prevalence for each sampling date, and examined temporal and spatial patterns by quantifying the annual level of ATV synchrony among populations using the intraclass correlation coefficient. Salamander populations were commonly infected with ATV. We observed no morbidity or mortality in these populations even as ATV prevalence values varied from 0 to 57%. Infection prevalence across the landscape was more similar within a given year than between years. There was no statistically significant spatial pattern in prevalence across the landscape. Our findings highlight the need to explore new hypotheses regarding the population level impact of these pathogens on amphibian communities.

Evaluating the Within-Host Dynamics of Ranavirus Infection with Mechanistic Disease Models and Experimental Data.

Mechanistic models are critical for our understanding of both within-host dynamics (i.e., pathogen replication and immune system processes) and among-host dynamics (i.e., transmission). Within-host models, however, are not often fit to experimental data, which can serve as a robust method of hypothesis testing and hypothesis generation. In this study, we use mechanistic models and empirical, time-series data of viral titer to better understand the replication of ranaviruses within their amphibian hosts and the immune dynamics that limit viral replication. Specifically, we fit a suite of potential models to our data, where each model represents a hypothesis about the interactions between viral replication and immune defense. Through formal model comparison, we find a parsimonious model that captures key features of our time-series data: The viral titer rises and falls through time, likely due to an immune system response, and that the initial viral dosage affects both the peak viral titer and the timing of the peak. Importantly, our model makes several predictions, including the existence of long-term viral infections, which can be validated in future studies.

Multiple causes of variable tick burdens on small-mammal hosts.

Blood meals by blacklegged ticks (Ixodes scapularis) on vertebrate hosts serve to transmit the agents of several zoonotic diseases, including Lyme disease, human babesiosis, and human granulocytic anaplasmosis, between host and tick. If ticks are aggregated on hosts, a small proportion of hosts may be responsible for most transmission events. Therefore, a key element in understanding and controlling the transmission of these pathogens is identifying the group(s) or individuals feeding a disproportionate number of ticks. Previous studies of tick burdens, however, have focused on differences in mean annual burdens between one or a few groups of hosts, ignoring both the strong seasonal dynamics of I. scapularis and their aggregation on hosts. We present a statistical modeling framework that predicts burdens on individual hosts throughout the year as a function of temporal-, site-, and individual-specific attributes, as well as the degree of aggregation in a negative binomial distribution. We then fit alternate versions of this model to an 11-year data set of I. scapularis burdens on white-footed mice (Peromyscus leucopus) and eastern chipmunks (Tamias striatus) to explore which factors are important to predicting tick burdens. We found that tick burdens are a complex function of many extrinsic and intrinsic factors, including seasonality. Specifically: (1) burdens on mice and chipmunks increased with densities of host-seeking ticks in a manner that suggests hosts become saturated. (2) Chipmunks draw larval ticks away from mice, which are efficient reservoirs of the Lyme disease bacterium, and mice draw nymphs away from chipmunks, which are key nymphal hosts. (3) While individual correlates were statistically important, the relationships were complex, and no group or correlate (sex, age, mass) could explain which hosts fed a disproportionate number of ticks. (4) Ticks were strongly aggregated on hosts within and across groups suggesting that some undiscovered quality of individual hosts was responsible for the aggregation. (5) Those individuals that fed more nymphs than expected, and are thus more likely to be infected with the Lyme disease agent, also tend to feed and infect more larvae than expected. Predicting which individuals those are is not yet possible.

Estimating reservoir competence of Borrelia burgdorferi hosts: prevalence and infectivity, sensitivity, and specificity.

Most vector-borne zoonotic pathogens are transmitted among several host species, but different species vary considerably in their importance to pathogen transmission, at least partially because they vary in their propensity to infect feeding vectors. This propensity is often called realized reservoir competence. Realized reservoir competence is the product of 1) the probability the individual host is infected, i.e., infection prevalence, and 2) the probability that if the host is infected, it will transmit the infection to a feeding vector, or infectivity. Prevalence varies in space and time, whereas infectivity may be a property of the host species. Both prevalence and infectivity are ecologically and epidemiologically important, but measuring them simultaneously is difficult. We present a probabilistic model that separately estimates host infection prevalence and infectivity from data on the infection status of vectors collected from individual hosts, data generally used to measure realized reservoir competence. We then consider how imperfect diagnostic tests (i.e., false negatives and positives) influence these probabilities-estimates of prevalence and infectivity are fairly robust to false negatives, but not to false positives. We thus extend the model to estimate the rate of false positives in order to improve estimates of prevalence and infectivity. We illustrate these methods by reanalyzing data from LoGiudice et al. (2003; Proc. Natl. Acad. Sci. U.S.A. 100: 567-571) on the reservoir competence of ten vertebrate hosts of Borrelia burgdorferi, the agent of Lyme disease. We find that these vertebrate hosts vary both in prevalence and infectivity and that both values are highly, positively correlated among species.

Evaluating the Importance of Environmental Persistence for Ranavirus Transmission and Epidemiology.

Viruses persist outside their hosts in a variety of forms, from naked virions to virus protected in sloughed tissues or carcasses, and for a range of times, all of which affect the likelihood and importance of transmission from the environment. This review synthesizes the literature on environmental persistence of viruses in the genus Ranavirus (family Iridoviridae), which are large double-stranded DNA viruses of ectothermic, often aquatic or semiaquatic vertebrates. Ranaviruses have been associated with mass mortality events in natural and captive settings around the world, and with population and community-wide declines in Europe. Early work suggested ranaviruses are environmentally robust and transmission from the environment should be common. More recent work has shown a large effect of temperature and microbial action on persistence times, although other aspects of the environment (e.g., water chemistry) and aquatic communities (e.g., zooplankton) may also be important. Ranaviruses may persist in the carcasses of animals that have died of infection, and so decomposing organisms and invertebrate scavengers may reduce these persistence times. The question is, do persistence times vary enough to promote or preclude substantial transmission from the environment. We built an epidemiological model with transmission from contacts, free virus in water, and carcasses, to explore the conditions in which environmental persistence could be important for ranavirus epidemiology. Based on prior work, we expected a substantial amount of transmission from the water and that longer persistence times would make this route of transmission dominant. However, neither water-borne nor transmission from carcasses played an important role in the simulated epidemics except under fairly restrictive conditions, such as when there were high rates of virus shedding or high rates of scavenging on highly infectious carcasses. While many aspects of environmental persistence of ranaviruses are being resolved by experiments, key parameters such as viral shedding rates are virtually unknown and will need to be empirically constrained if we are to determine whether environmental persistence and transmission from the environment are essential or insignificant features of Ranavirus epidemiology. We conclude by emphasizing the need to place environmental persistence research in an epidemiological framework.

Susceptibility of the endangered California tiger salamander, Ambystoma californiense, to ranavirus infection.

Emerging infectious diseases are implicated in the declines and extinctions of amphibians worldwide. Ranaviruses in the family Iridoviridae are a global concern and have caused amphibian die-offs in wild populations in North America, Europe, South America, and in commercial populations in Asia and South America. The movement of amphibians for bait, food, pets, and research provides a route for the introduction of ranaviruses into naive and potentially endangered species. In this report, we demonstrate that the California tiger salamander, Ambystoma californiense, is susceptible to Ambystoma tigrinum virus (ATV). This virus has not been previously reported in California tiger salamander, but observed mortality in experimentally infected animals suggests that California tiger salamander populations could be adversely affected by an ATV introduction.

From fish to frogs and beyond: Impact and host range of emergent ranaviruses.

Ranaviruses are pathogens of ectothermic vertebrates, including amphibians. We reviewed patterns of host range and virulence of ranaviruses in the context of virus genotype and postulate that patterns reflect significant variation in the historical and current host range of three groups of Ranavirus: FV3-like, CMTV-like and ATV-like ranaviruses. Our synthesis supports previous hypotheses about host range and jumps: FV3s are amphibian specialists, while ATVs are predominantly fish specialists that switched once to caudate amphibians. The most recent common ancestor of CMTV-like ranaviruses and FV3-like forms appears to have infected amphibians but CMTV-like ranaviruses may circulate in both amphibian and fish communities independently. While these hypotheses are speculative, we hope that ongoing efforts to describe ranavirus genetics, increased surveillance of host species and targeted experimental assays of susceptibility to infection and/or disease will facilitate better tests of the importance of hypothetical evolutionary drivers of ranavirus virulence and host range.

Evaluating environmental DNA-based quantification of ranavirus infection in wood frog populations.

A variety of challenges arise when monitoring wildlife populations for disease. Sampling tissues can be invasive to hosts, and obtaining sufficient sample sizes can be expensive and time-consuming, particularly for rare species and when pathogen prevalence is low. Environmental DNA (eDNA)-based detection of pathogens is an alternative approach to surveillance for aquatic communities that circumvents many of these issues. Ranaviruses are emerging pathogens of ectothermic vertebrates linked to die-offs of amphibian populations. Detecting ranavirus infections is critical, but nonlethal methods have the above issues and are prone to false negatives. We report on the feasibility and effectiveness of eDNA-based ranavirus detection in the field. We compared ranavirus titres in eDNA samples collected from pond water to titres in wood frog (Lithobates sylvaticus; n = 5) tadpoles in sites dominated by this one species (n = 20 pond visits). We examined whether ranavirus DNA can be detected in eDNA from pond water when infections are present in the pond and if viral titres detected in eDNA samples correlate with the prevalence or intensity of ranavirus infections in tadpoles. With three 250 mL water samples, we were able to detect the virus in all visits with infected larvae (0.92 diagnostic sensitivity). Also, we found a strong relationship between the viral eDNA titres and titres in larval tissues. eDNA titres increased prior to observed die-offs and declined afterwards, and were two orders of magnitude higher in ponds with a die-off. Our results suggest that eDNA is useful for detecting ranavirus infections in wildlife and aquaculture.

Climate change and Ixodes tick-borne diseases of humans.

The evidence that climate warming is changing the distribution of Ixodes ticks and the pathogens they transmit is reviewed and evaluated. The primary approaches are either phenomenological, which typically assume that climate alone limits current and future distributions, or mechanistic, asking which tick-demographic parameters are affected by specific abiotic conditions. Both approaches have promise but are severely limited when applied separately. For instance, phenomenological approaches (e.g. climate envelope models) often select abiotic variables arbitrarily and produce results that can be hard to interpret biologically. On the other hand, although laboratory studies demonstrate strict temperature and humidity thresholds for tick survival, these limits rarely apply to field situations. Similarly, no studies address the influence of abiotic conditions on more than a few life stages, transitions or demographic processes, preventing comprehensive assessments. Nevertheless, despite their divergent approaches, both mechanistic and phenomenological models suggest dramatic range expansions of Ixodes ticks and tick-borne disease as the climate warms. The predicted distributions, however, vary strongly with the models' assumptions, which are rarely tested against reasonable alternatives. These inconsistencies, limited data about key tick-demographic and climatic processes and only limited incorporation of non-climatic processes have weakened the application of this rich area of research to public health policy or actions. We urge further investigation of the influence of climate on vertebrate hosts and tick-borne pathogen dynamics. In addition, testing model assumptions and mechanisms in a range of natural contexts and comparing their relative importance as competing models in a rigorous statistical framework will significantly advance our understanding of how climate change will alter the distribution, dynamics and risk of tick-borne disease.