Rapid adaptation to a novel pathogen through disease tolerance in a wild songbird.

Animal hosts can adapt to emerging infectious disease through both disease resistance, which decreases pathogen numbers, and disease tolerance, which limits damage during infection without limiting pathogen replication. Both resistance and tolerance mechanisms can drive pathogen transmission dynamics. However, it is not well understood how quickly host tolerance evolves in response to novel pathogens or what physiological mechanisms underlie this defense. Using natural populations of house finches (Haemorhous mexicanus) across the temporal invasion gradient of a recently emerged bacterial pathogen (Mycoplasma gallisepticum), we find rapid evolution of tolerance (<25 years). In particular, populations with a longer history of MG endemism have less pathology but similar pathogen loads compared with populations with a shorter history of MG endemism. Further, gene expression data reveal that more-targeted immune responses early in infection are associated with tolerance. These results suggest an important role for tolerance in host adaptation to emerging infectious diseases, a phenomenon with broad implications for pathogen spread and evolution.

Monarch Butterfly Ecology, Behavior, and Vulnerabilities in North Central United States Agricultural Landscapes.

The North American monarch butterfly (Danaus plexippus) is a candidate species for listing under the Endangered Species Act. Multiple factors are associated with the decline in the eastern population, including the loss of breeding and foraging habitat and pesticide use. Establishing habitat in agricultural landscapes of the North Central region of the United States is critical to increasing reproduction during the summer. We integrated spatially explicit modeling with empirical movement ecology and pesticide toxicology studies to simulate population outcomes for different habitat establishment scenarios. Because of their mobility, we conclude that breeding monarchs in the North Central states should be resilient to pesticide use and habitat fragmentation. Consequently, we predict that adult monarch recruitment can be enhanced even if new habitat is established near pesticide-treated crop fields. Our research has improved the understanding of monarch population dynamics at the landscape scale by examining the interactions among monarch movement ecology, habitat fragmentation, and pesticide use.

Repeated letters increase the ambiguity of strings: Evidence from identification, priming and same-different tasks.

Letters are often repeated in words in many languages. The present work explored the mechanisms underlying processing of repeated and unique letters in strings across three experimental paradigms. In a 2AFC perceptual identification task, the insertion but not the deletion of a letter was harder to detect when it was repeated than when it was unique (Exp. 1). In a masked primed same-different task, deletion primes produced the same priming effect regardless of deletion type (repeated, unique; Exp. 2), but insertion primes were more effective when the additional inserted letter created a repetition than when it did not (Exp. 3). In a same-different perceptual identification task, foils created by modifying a repetition, by either repeating the wrong letter or substituting a repeated letter, were harder to reject than foils created by modifying unique letters (Exp. 4). Thus, repetition effects were task-dependent. Since considering representations alone would suggest repetition effects would always occur or never occur, this indicates the importance of modelling task-specific processes. The similarity calculations embedded in the Overlap Model (Gomez et al., 2008) appeared to always predict a repetition effect, but its decision rule for the task of Experiment 1 allowed it to predict the asymmetry between insertions and deletions. In the Letters in Time and Retinotopic Space (LTRS; Adelman, 2011) model, repetition effects arise only from briefly presented stimuli as their perception is incomplete. It was therefore consistent with Experiments 2-4 but required a task-specific response bias to account for the insertion-deletion asymmetry of Experiment 1.

Disease tolerance alters host competence in a wild songbird.

Individuals can express a range of disease phenotypes during infection, with important implications for epidemics. Tolerance, in particular, is a host response that minimizes the per-pathogen fitness costs of infection. Because tolerant hosts show milder clinical signs and higher survival, despite similar pathogen burdens, their potential for prolonged pathogen shedding may facilitate the spread of pathogens. To test this, we simulated outbreaks of mycoplasmal conjunctivitis in house finches, asking how the speed of transmission varied with tissue-specific and behavioural components of tolerance, milder conjunctivitis and anorexia for a given pathogen load, respectively. Because tissue-specific tolerance hinders pathogen deposition onto bird feeders, important transmission hubs, we predicted it would slow transmission. Because behavioural tolerance should increase interactions with bird feeders, we predicted it would speed transmission. Our findings supported these predictions, suggesting that variation in tolerance could help identify individuals most likely to transmit pathogens.

Exploratory behavior is linked to stress physiology and social network centrality in free-living house finches (Haemorhous mexicanus).

Animal personality has been linked to individual variation in both stress physiology and social behaviors, but few studies have simultaneously examined covariation between personality traits, stress hormone levels, and behaviors in free-living animals. We investigated relationships between exploratory behavior (one aspect of animal personality), stress physiology, and social and foraging behaviors in wild house finches (Haemorhous mexicanus). We conducted novel environment assays after collecting samples of baseline and stress-induced plasma corticosterone concentrations from a subset of house finches. We then fitted individuals with Passive Integrated Transponder tags and monitored feeder use and social interactions at radio-frequency identification equipped bird feeders. First, we found that individuals with higher baseline corticosterone concentrations exhibit more exploratory behaviors in a novel environment. Second, more exploratory individuals interacted with more unique conspecifics in the wild, though this result was stronger for female than for male house finches. Third, individuals that were quick to begin exploring interacted more frequently with conspecifics than slow-exploring individuals. Finally, exploratory behaviors were unrelated to foraging behaviors, including the amount of time spent on bird feeders, a behavior previously shown to be predictive of acquiring a bacterial disease that causes annual epidemics in house finches. Overall, our results indicate that individual differences in exploratory behavior are linked to variation in both stress physiology and social network traits in free-living house finches. Such covariation has important implications for house finch ecology, as both traits can contribute to fitness in the wild.

Characterization of unilateral conjunctival inoculation with Mycoplasma gallisepticum in house finches.

House finches in much of the continental United States experience annual epidemics of mycoplasmal conjunctivitis, caused by the bacterial pathogen Mycoplasma gallisepticum (MG). Although evidence suggests that natural infections typically begin unilaterally, experimental inoculations of songbirds with MG to date have all been administered bilaterally. Furthermore, studies of free-living finches find more severe clinical signs of mycoplasmal conjunctivitis in left versus right eyes, but the mechanisms underlying this side bias remain unknown. Here, we characterized unilateral inoculation of house finches with MG, and tested whether differential susceptibility of left versus right conjunctiva explains the side bias in disease severity of free-living finches. We directly inoculated house finches in either the left or right conjunctiva and characterized resulting disease severity and pathogen load throughout the course of infection. As expected, unilateral inoculation resulted in significantly more severe conjunctivitis, as well as higher conjunctival bacterial loads, on whichever side (left or right) birds were directly inoculated. However, in 55% of cases, unilateral inoculations resulted in bilateral disease, and in 85% cases there was evidence of bilateral infection. The overall severity of disease did not differ for birds inoculated in the left versus right conjunctiva, suggesting that physiological differences between the conjunctivae cannot explain the side bias in disease severity of free-living birds. Instead, laterality in exposure, perhaps due to feeding handedness, likely explains the detected field patterns. RESEARCH HIGHLIGHTS House finches show more severe disease in the directly inoculated conjunctiva. Unilateral inoculations lead to high rates of bilateral infection and disease. Overall disease severity does not differ for the left- or right-inoculated conjunctiva. Laterality in exposure likely explains the left-side bias in natural infections.

Tolerance of infection: A role for animal behavior, potential immune mechanisms, and consequences for parasite transmission.

Infected organisms can resist or tolerate infection, with tolerance of infection defined as minimizing per-parasite reductions in fitness. Although tolerance is well studied in plants, researchers have only begun to probe the mechanisms and transmission consequences of tolerance in animals. Here we suggest that research on tolerance in animals would benefit from explicitly incorporating behavior as a component of tolerance, given the importance of behavior for host fitness and parasite transmission. We propose two distinct manifestations of tolerance in animals: tissue-specific tolerance, which minimizes fitness losses due to tissue damage during infection, and behavioral tolerance, which minimizes fitness losses by maintaining normal, fitness-enhancing behaviors during infection. Here we briefly review one set of potential immune mechanisms underlying both responses in vertebrate animals: inflammation and its associated signaling molecules. Inflammatory responses, including broadly effective resistance mechanisms like the production of reactive oxygen species, can incur severe costs in terms of damage to a host's own tissues, thereby reducing tissue-specific tolerance. In addition, signaling molecules involved in these responses facilitate stereotypical behavioral changes during infection, which include lethargy and anorexia, reducing normal behaviors and behavioral tolerance. We consider how tissue-specific and behavioral tolerance may vary independently or in conjunction and outline potential consequences of such covariation for the transmission of infectious diseases. We put forward the distinction between tissue-specific and behavioral tolerance not as a definitive framework, but to help stimulate and broaden future research by considering animal behavior as intimately linked to the mechanisms and consequences of tolerance in animals.

Development and validation of a house finch interleukin-1ß (HfIL-1ß) ELISA system.

BACKGROUND: A unique clade of the bacterium Mycoplasma gallisepticum (MG), which causes chronic respiratory disease in poultry, has resulted in annual epidemics of conjunctivitis in North American house finches since the 1990s. Currently, few immunological tools have been validated for this songbird species. Interleukin-1ß (IL-1ß) is a prototypic multifunctional cytokine and can affect almost every cell type during Mycoplasma infection. The overall goal of this study was to develop and validate a direct ELISA assay for house finch IL-1ß (HfIL-1ß) using a cross-reactive chicken antibody. METHODS: A direct ELISA approach was used to develop this system using two different coating methods, carbonate and dehydration. In both methods, antigens (recombinant HfIL-1b or house finch plasma) were serially diluted in carbonate-bicarbonate coating buffer and either incubated at 4 °C overnight or at 60 °C on a heating block for 2 hr. To generate the standard curve, rHfIL-1b protein was serially diluted at 0, 3, 6, 9, 12, 15, 18, 21, and 24 ng/mL. Following blocking and washing, anti-chicken IL-1b polyclonal antibody was added, plates were later incubated with detecting antibodies, and reactions developed with tetramethylbenzidine solution. RESULTS: A commercially available anti-chicken IL-1ß (ChIL-1ß) polyclonal antibody (pAb) cross-reacted with house finch plasma IL-1ß as well as bacterially expressed recombinant house finch IL-1ß (rHfIL-1ß) in immunoblotting assays. In a direct ELISA system, rHfIL-1ß could not be detected by an anti-ChIL-1ß pAb when the antigen was coated with carbonate-bicarbonate buffer at 4°C overnight. However, rHfIL-1ß was detected by the anti-ChIL-1ß pAb when the antigen was coated using a dehydration method by heat (60°C). Using the developed direct ELISA for HfIL-1ß with commercial anti-ChIL-1ß pAb, we were able to measure plasma IL-1ß levels from house finches. CONCLUSIONS: Based on high amino acid sequence homology, we hypothesized and demonstrated cross-reactivity of anti-ChIL-1ß pAb and HfIL-1ß. Then, we developed and validated a direct ELISA system for HfIL-1ß using a commercial anti-ChIL-1ß pAb by measuring plasma HfIL-1ß in house finches.

A robust preference for cheap-and-easy strategies over reliable strategies when verifying personal memories.

People depend on various sources of information when trying to verify their autobiographical memories. Yet recent research shows that people prefer to use cheap-and-easy verification strategies, even when these strategies are not reliable. We examined the robustness of this cheap strategy bias, with scenarios designed to encourage greater emphasis on source reliability. In three experiments, subjects described real (Experiments 1 and 2) or hypothetical (Experiment 3) autobiographical events, and proposed strategies they might use to verify their memories of those events. Subjects also rated the reliability, cost, and the likelihood that they would use each strategy. In line with previous work, we found that the preference for cheap information held when people described how they would verify childhood or recent memories (Experiment 1), personally important or trivial memories (Experiment 2), and even when the consequences of relying on incorrect information could be significant (Experiment 3). Taken together, our findings fit with an account of source monitoring in which the tendency to trust one's own autobiographical memories can discourage people from systematically testing or accepting strong disconfirmatory evidence.

Changes in corticosterone concentrations and behavior during Mycoplasma gallisepticum infection in house finches (Haemorhous mexicanus).

Glucocorticoid stress hormones are important for energy mobilization as well as regulation of the immune system, and thus these hormones are particularly likely to both influence and respond to pathogen infection in vertebrates. In this study, we examined how the glucocorticoid stress response in house finches (Haemorhous mexicanus) interacts with experimental infection of the naturally-occurring bacterial pathogen, Mycoplasma gallisepticum (MG). We also investigated whether infection-induced concentrations of corticosterone (CORT), the primary glucocorticoid in birds, were associated with the expression of sickness behavior, the lethargy typically observed in vertebrates early in infection. We found that experimental infection with MG resulted in significantly higher CORT levels on day 5 post-infection, but this effect appeared to be limited to female house finches only. Regardless of sex, infected individuals with greater disease severity had the highest CORT concentrations on day 5 post-infection. House finches exposed to MG exhibited behavioral changes, with infected birds having significantly lower activity levels than sham-inoculated individuals. However, CORT concentrations and the extent of sickness behaviors exhibited among infected birds were not associated. Finally, pre-infection CORT concentrations were associated with reduced inflammation and pathogen load in inoculated males, but not females. Our results suggest that the house finch glucocorticoid stress response may both influence and respond to MG infection in sex-specific ways, but because we had a relatively low sample size of males, future work should confirm these patterns. Finally, manipulative experiments should be performed to test whether the glucocorticoid stress response acts as a brake on the inflammatory response associated with MG infection in house finches.

Feeder use predicts both acquisition and transmission of a contagious pathogen in a North American songbird.

Individual heterogeneity can influence the dynamics of infectious diseases in wildlife and humans alike. Thus, recent work has sought to identify behavioural characteristics that contribute disproportionately to individual variation in pathogen acquisition (super-receiving) or transmission (super-spreading). However, it remains unknown whether the same behaviours enhance both acquisition and transmission, a scenario likely to result in explosive epidemics. Here, we examined this possibility in an ecologically relevant host-pathogen system: house finches and their bacterial pathogen, Mycoplasma gallisepticum, which causes severe conjunctivitis. We examined behaviours likely to influence disease acquisition (feeder use, aggression, social network affiliations) in an observational field study, finding that the time an individual spends on bird feeders best predicted the risk of conjunctivitis. To test whether this behaviour also influences the likelihood of transmitting M. gallisepticum, we experimentally inoculated individuals based on feeding behaviour and tracked epidemics within captive flocks. As predicted, transmission was fastest when birds that spent the most time on feeders initiated the epidemic. Our results suggest that the same behaviour underlies both pathogen acquisition and transmission in this system and potentially others. Identifying individuals that exhibit such behaviours is critical for disease management.

Repeated stressors in adulthood increase the rate of biological ageing.

BACKGROUND: Individuals of the same age can differ substantially in the degree to which they have accumulated tissue damage, akin to bodily wear and tear, from past experiences. This accumulated tissue damage reflects the individual's biological age and may better predict physiological and behavioural performance than the individual's chronological age. However, at present it remains unclear how to reliably assess biological age in individual wild vertebrates. METHODS: We exposed hand-raised adult Eurasian blackbirds (Turdus merula) to a combination of repeated immune and disturbance stressors for over one year to determine the effects of chronic stress on potential biomarkers of biological ageing including telomere shortening, oxidative stress load, and glucocorticoid hormones. We also assessed general measures of individual condition including body mass and locomotor activity. RESULTS: By the end of the experiment, stress-exposed birds showed greater decreases in telomere lengths. Stress-exposed birds also maintained higher circulating levels of oxidative damage compared with control birds. Other potential biomarkers such as concentrations of antioxidants and glucocorticoid hormone traits showed greater resilience and did not differ significantly between treatment groups. CONCLUSIONS: The current data demonstrate that repeated exposure to experimental stressors affects the rate of biological ageing in adult Eurasian blackbirds. Both telomeres and oxidative damage were affected by repeated stress exposure and thus can serve as blood-derived biomarkers of biological ageing.

Individual differences in reading aloud: a mega-study, item effects, and some models.

Normal individual differences are rarely considered in the modelling of visual word recognition--with item response time effects and neuropsychological disorders being given more emphasis--but such individual differences can inform and test accounts of the processes of reading. We thus had 100 participants read aloud words selected to assess theoretically important item response time effects on an individual basis. Using two major models of reading aloud--DRC and CDP+--we estimated numerical parameters to best model each individual's response times to see if this would allow the models to capture the effects, individual differences in them and the correlations among these individual differences. It did not. We therefore created an alternative model, the DRC-FC, which successfully captured more of the correlations among individual differences, by modifying the locus of the frequency effect. Overall, our analyses indicate that (i) even after accounting for individual differences in general speed, several other individual difference in reading remain significant; and (ii) these individual differences provide critical tests of models of reading aloud. The database thus offers a set of important constraints for future modelling of visual word recognition, and is a step towards integrating such models with other knowledge about individual differences in reading.

Linking social environment and stress physiology in feral mares (Equus caballus): group transfers elevate fecal cortisol levels.

Feral horses (Equus caballus) have a complex social structure, the stability of which is important to their overall health. Behavioral and demographic research has shown that decreases in group (or band) stability reduce female fitness, but the potential effects on the physiological stress response have not been demonstrated. To fully understand how band stability affects group-member fitness, we need to understand not only behavioral and demographic, but also physiological consequences of decreases to that stability. We studied group changes in feral mares (an activity that induces instability, including both male and female aggression) on Shackleford Banks, NC. We found that mares in the midst of changing groups exhibit increased fecal cortisol levels. In addition, mares making more group transfers show higher levels of cortisol two weeks post-behavior. These results offer insights into how social instability is integrated into an animal's physiological phenotype. In addition, our results have important implications for feral horse management. On Shackleford Banks, mares contracepted with porcine zona pellucida (PZP) make approximately 10 times as many group changes as do untreated mares. Such animals may therefore be at higher risk of chronic stress. These results support the growing consensus that links between behavior and physiological stress must be taken into account when managing for healthy, functional populations.

A behavioral database for masked form priming.

Reading involves a process of matching an orthographic input with stored representations in lexical memory. The masked priming paradigm has become a standard tool for investigating this process. Use of existing results from this paradigm can be limited by the precision of the data and the need for cross-experiment comparisons that lack normal experimental controls. Here, we present a single, large, high-precision, multicondition experiment to address these problems. Over 1,000 participants from 14 sites responded to 840 trials involving 28 different types of orthographically related primes (e.g., castfe-CASTLE) in a lexical decision task, as well as completing measures of spelling and vocabulary. The data were indeed highly sensitive to differences between conditions: After correction for multiple comparisons, prime type condition differences of 2.90 ms and above reached significance at the 5% level. This article presents the method of data collection and preliminary findings from these data, which included replications of the most widely agreed-upon differences between prime types, further evidence for systematic individual differences in susceptibility to priming, and new evidence regarding lexical properties associated with a target word's susceptibility to priming. These analyses will form a basis for the use of these data in quantitative model fitting and evaluation and for future exploration of these data that will inform and motivate new experiments.

Varying conjunctival immune response adaptations of house finch populations to a rapidly evolving bacterial pathogen.

Pathogen adaptations during host-pathogen co-evolution can cause the host balance between immunity and immunopathology to rapidly shift. However, little is known in natural disease systems about the immunological pathways optimised through the trade-off between immunity and self-damage. The evolutionary interaction between the conjunctival bacterial infection Mycoplasma gallisepticum (MG) and its avian host, the house finch (Haemorhous mexicanus), can provide insights into such adaptations in immune regulation. Here we use experimental infections to reveal immune variation in conjunctival tissue for house finches captured from four distinct populations differing in the length of their co-evolutionary histories with MG and their disease tolerance (defined as disease severity per pathogen load) in controlled infection studies. To differentiate contributions of host versus pathogen evolution, we compared house finch responses to one of two MG isolates: the original VA1994 isolate and a more evolutionarily derived one, VA2013. To identify differential gene expression involved in initiation of the immune response to MG, we performed 3'-end transcriptomic sequencing (QuantSeq) of samples from the infection site, conjunctiva, collected 3-days post-infection. In response to MG, we observed an increase in general pro-inflammatory signalling, as well as T-cell activation and IL17 pathway differentiation, associated with a decrease in the IL12/IL23 pathway signalling. The immune response was stronger in response to the evolutionarily derived MG isolate compared to the original one, consistent with known increases in MG virulence over time. The host populations differed namely in pre-activation immune gene expression, suggesting population-specific adaptations. Compared to other populations, finches from Virginia, which have the longest co-evolutionary history with MG, showed significantly higher expression of anti-inflammatory genes and Th1 mediators. This may explain the evolution of disease tolerance to MG infection in VA birds. We also show a potential modulating role of BCL10, a positive B- and T-cell regulator activating the NFKB signalling. Our results illuminate potential mechanisms of house finch adaptation to MG-induced immunopathology, contributing to understanding of the host evolutionary responses to pathogen-driven shifts in immunity-immunopathology trade-offs.

Prior exposure to pathogens augments host heterogeneity in susceptibility and has key epidemiological consequences.

Pathogen epidemics are key threats to human and wildlife health. Across systems, host protection from pathogens following initial exposure is often incomplete, resulting in recurrent epidemics through partially-immune hosts. Variation in population-level protection has important consequences for epidemic dynamics, but whether acquired protection influences host heterogeneity in susceptibility and its epidemiological consequences remains unexplored. We experimentally investigated whether prior exposure (none, low-dose, or high-dose) to a bacterial pathogen alters host heterogeneity in susceptibility among songbirds. Hosts with no prior pathogen exposure had little variation in protection, but heterogeneity in susceptibility was significantly augmented by prior pathogen exposure, with the highest variability detected in hosts given high-dose prior exposure. An epidemiological model parameterized with experimental data found that heterogeneity in susceptibility from prior exposure more than halved epidemic sizes compared with a homogeneous population with identical mean protection. However, because infection-induced mortality was also greatly reduced in hosts with prior pathogen exposure, reductions in epidemic size were smaller than expected in hosts with prior exposure. These results highlight the importance of variable protection from prior exposure and/or vaccination in driving host heterogeneity and epidemiological dynamics.

House finch populations differ in early inflammatory signaling and pathogen tolerance at the peak of Mycoplasma gallisepticum infection.

Host individuals and populations often vary in their responses to infection, with direct consequences for pathogen spread and evolution. While considerable work has focused on the mechanisms underlying differences in resistance-the ability to kill pathogens-we know little about the mechanisms underlying tolerance-the ability to minimize fitness losses per unit pathogen. Here, we examine patterns and mechanisms of tolerance between two populations of house finches (Haemorhous [formerly Carpodacus] mexicanus) with different histories with the bacterial pathogen Mycoplasma gallisepticum (MG). After infection in a common environment, we assessed two metrics of pathology, mass loss and eye lesion severity, as proxies for fitness. We calculated tolerance using two methods, one based on pathology and pathogen load at the peak of infection (point tolerance) and the other based on the integrals of these metrics over time (range tolerance). Alabama birds, which have a significantly longer history of exposure to MG, showed more pronounced point tolerance than Arizona birds, while range tolerance did not differ between populations. Alabama birds also displayed lower inflammatory cytokine signaling and lower fever early in infection. These results suggest that differences in inflammatory processes, which can significantly damage host tissues, may contribute to variation in tolerance among house finch individuals and populations. Such variation can affect pathogen spread and evolution in ways not predictable by resistance alone and sheds light on the costs and benefits of inflammation in wild animals.

A free-ranging, feral mare equus caballus affords similar maternal care to her genetic and adopted offspring.

Adoption of nongenetic offspring occurs in a variety of species but is rare in equids. We report a case of adoption by a free-ranging, feral mare Equus caballus and compare the maternal care received by her genetic offspring (born 1995) to that of her adopted offspring (born 1996) for the first 30 weeks of development. We compare five measures of care: (1) total time spent suckling, (2) mare aggression during suckling, (3) number of mare-terminated suckling bouts, (4) contact maintenance, and (5) mare-foal distance. For most behaviors, we detected no difference in the mare's treatment of the two foals; however, mare-foal distance was greater for the genetic offspring. We compare hypotheses regarding the reasons for adoption, offering postpartum physiological state as a potential driver.

Deposition of pathogenic Mycoplasma gallisepticum onto bird feeders: host pathology is more important than temperature-driven increases in food intake.

Although ambient temperature has diverse effects on disease dynamics, few studies have examined how temperature alters pathogen transmission by changing host physiology or behaviour. Here, we test whether reducing ambient temperature alters host foraging, pathology and the potential for fomite transmission of the bacterial pathogen Mycoplasma gallisepticum (MG), which causes seasonal outbreaks of severe conjunctivitis in house finches (Haemorhous mexicanus). We housed finches at temperatures within or below the thermoneutral zone to manipulate food intake by altering energetic requirements of thermoregulation. We predicted that pathogen deposition on bird feeders would increase with temperature-driven increases in food intake and with conjunctival pathology. As expected, housing birds below the thermoneutral zone increased food consumption. Despite this difference, pathogen deposition on feeders did not vary across temperature treatments. However, pathogen deposition increased with conjunctival pathology, independently of temperature and pathogen load, suggesting that MG could enhance its transmission by increasing virulence. Our results suggest that in this system, host physiological responses are more important for transmission potential than temperature-dependent alterations in feeding. Understanding such behavioural and physiological contributions to disease transmission is critical to linking individual responses to climate with population-level disease dynamics.