Shifts in population density centers of a hibernating mammal driven by conflicting effects of climate change and disease.

Populations wax and wane over time in response to an organism's interactions with abiotic and biotic forces. Numerous studies demonstrate that fluctuations in local populations can lead to shifts in relative population densities across the geographic range of a species over time. Fewer studies attempt to disentangle the causes of such shifts. Over four decades (1983-2022), we monitored populations of hibernating Indiana bats (Myotis sodalis) in two areas separated by ~110 km. The number of bats hibernating in the northern area increased from 1983 to 2011, while populations in the southern area remained relatively constant. We used simulation models and long-term weather data to demonstrate the duration of time bats must rely on stored fat during hibernation has decreased in both areas over that period, but at a faster rate in the northern area. Likewise, increasing autumn and spring temperatures shortened the periods of sporadic prey (flying insect) availability at the beginning and end of hibernation. Climate change thus increased the viability of northern hibernacula for an increasing number of bats by decreasing energetic costs of hibernation. Then in 2011, white-nose syndrome (WNS), a disease of hibernating bats that increases energetic costs of hibernation, was detected in the area. From 2011 to 2022, the population rapidly decreased in the northern area and increased in the southern area, completely reversing the northerly shift in population densities associated with climate change. Energy balance during hibernation is the singular link explaining the northerly shift under a changing climate and the southerly shift in response to a novel disease. Continued population persistence suggests that bats may mitigate many impacts of WNS by hibernating farther south, where insects are available longer each year.

Plant pathogens provide clues to the potential origin of bat white-nose syndrome Pseudogymnoascus destructans.

White-nose syndrome has killed millions of bats, yet both the origins and infection strategy of the causative fungus, Pseudogymnoascus destructans, remain elusive. We provide evidence for a novel hypothesis that P. destructans emerged from plant-associated fungi and retained invasion strategies affiliated with fungal pathogens of plants. We demonstrate that P. destructans invades bat skin in successive biotrophic and necrotrophic stages (hemibiotrophic infection), a mechanism previously only described in plant fungal pathogens. Further, the convergence of hyphae at hair follicles suggests nutrient tropism. Tropism, biotrophy, and necrotrophy are often associated with structures termed appressoria in plant fungal pathogens; the penetrating hyphae produced by P. destructans resemble appressoria. Finally, we conducted a phylogenomic analysis of a taxonomically diverse collection of fungi. Despite gaps in genetic sampling of prehistoric and contemporary fungal species, we estimate an 88% probability the ancestral state of the clade containing P. destructans was a plant-associated fungus.

Experimental inoculation trial to determine the effects of temperature and humidity on White-nose Syndrome in hibernating bats.

Disease results from interactions among the host, pathogen, and environment. Inoculation trials can quantify interactions among these players and explain aspects of disease ecology to inform management in variable and dynamic natural environments. White-nose Syndrome, a disease caused by the fungal pathogen, Pseudogymnoascus destructans (Pd), has caused severe population declines of several bat species in North America. We conducted the first experimental infection trial on the tri-colored bat, Perimyotis subflavus, to test the effect of temperature and humidity on disease severity. We also tested the effects of temperature and humidity on fungal growth and persistence on substrates. Unexpectedly, only 37% (35/95) of bats experimentally inoculated with Pd at the start of the experiment showed any infection response or disease symptoms after 83 days of captive hibernation. There was no evidence that temperature or humidity influenced infection response. Temperature had a strong effect on fungal growth on media plates, but the influence of humidity was more variable and uncertain. Designing laboratory studies to maximize research outcomes would be beneficial given the high costs of such efforts and potential for unexpected outcomes. Understanding the influence of microclimates on host-pathogen interactions remains an important consideration for managing wildlife diseases, particularly in variable environments.

Heterothermy as a mechanism to offset energetic costs of environmental and homeostatic perturbations.

Environmental and biotic pressures impose homeostatic costs on all organisms. The energetic costs of maintaining high body temperatures (Tb) render endotherms sensitive to pressures that increase foraging costs. In response, some mammals become more heterothermic to conserve energy. We measured Tb in banner-tailed kangaroo rats (Dipodomys spectabilis) to test and disentangle the effects of air temperature and moonlight (a proxy for predation risk) on thermoregulatory homeostasis. We further perturbed homeostasis in some animals with chronic corticosterone (CORT) via silastic implants. Heterothermy increased across summer, consistent with the predicted effect of lunar illumination (and predation), and in the direction opposite to the predicted effect of environmental temperatures. The effect of lunar illumination was also evident within nights as animals maintained low Tb when the moon was above the horizon. The pattern was accentuated in CORT-treated animals, suggesting they adopted an even further heightened risk-avoidance strategy that might impose reduced foraging and energy intake. Still, CORT-treatment did not affect body condition over the entire study, indicating kangaroo rats offset decreases in energy intake through energy savings associated with heterothermy. Environmental conditions receive the most attention in studies of thermoregulatory homeostasis, but we demonstrated here that biotic factors can be more important and should be considered in future studies.

Temperature alone is insufficient to understand hibernation energetics.

Energy conservation has long been a focal point in hibernation research. A long-standing assumption is that ambient temperature (Ta) largely defines the rate of energy expenditure because of well-known relationships between Ta, metabolic rate and frequency of arousal from torpor. Body condition and humidity also affect energy expenditure but are usually considered secondary factors. We held tricolored bats (Perimyotis subflavus) in captivity under multiple environmental conditions to directly compare the importance of Ta, fat mass and humidity for hibernation energy expenditure. Fat mass was the best predictor of female mass loss, followed by Ta and humidity. However, males had less fat and adopted a more energetically conservative hibernation strategy. Our results demonstrate that understanding the evolution of behavior, physiology and ecology of hibernation requires disentangling the relative contributions of multiple drivers of hibernation energetics, and that Ta is not always the most important factor driving energy expenditure.

High Body Temperature is an Unlikely Cause of High Viral Tolerance in Bats.

The global SARS-CoV-2 pandemic and the role of bats in zoonotic spillover have renewed interest in the flight-as-fever hypothesis, which posits that high body temperatures experienced by bats during flight contribute to their high viral tolerance. We argue that flight-as-fever is unlikely to explain why bats harbor more viruses than other mammals on the basis of two lines of reasoning. First, flight temperatures reported in the literature overestimate true flight temperatures because of methodologic limitations. Second, body temperatures in bats are only high relative to humans, and not relative to many other mammals. We provide examples of mammals from diverse habitats to show that temperatures in excess of 40 C during activity are quite common in species with lower viral diversity than bats. We caution scientists against stating the flight-as-fever hypothesis as unquestioned truth, as has repeatedly occurred in the popular media in the wake of the SARS-CoV-2 pandemic.

An oversimplification of physiological principles leads to flawed macroecological analyses.

Macrophysiological analyses are useful to predict current and future range limits and improve our understanding of endotherm macroecology, but such analyses too often rely on oversimplifications of endothermic thermoregulatory and energetic physiology, which lessens their applicability. We detail some of the major issues with macrophysiological analyses based on the classic Scholander-Irving model of endotherm energetics in the hope that it will encourage other research teams to more appropriately integrate physiology into macroecological analyses.

A Brief Introduction to Methods for Describing Body Temperature in Endotherms.

Researchers commonly measure body, orifice, or skin temperature (collectively referred to as body temperature [Tb] herein) of endothermic animals in biomedical, physiological, evolutionary, and ecological studies. However, comparing Tb among species or placing a single study in context is challenging because there is no single, standard method to describe and synthesize Tb data of endotherms. A variety of metrics are available, and each has strengths and weaknesses appropriate for answering different types of questions. Importantly, choosing the wrong metric to address the question posed can lead to misinterpretations and misleading presentation of Tb data. Here I review standard metrics used to describe central tendencies and variation in Tb of endothermic species, focusing on important strengths and weaknesses and suggested questions to be addressed using each metric. One of the most common mistakes in analyzing Tb data is mismatching the analytical metric and the question being asked, so, ultimately, individual researchers need to determine which is most appropriate for addressing their question based on the implications of using each metric.

An experimental test of the allotonic frequency hypothesis to isolate the effects of light pollution on bat prey selection.

Artificial lights may be altering interactions between bats and moth prey. According to the allotonic frequency hypothesis (AFH), eared moths are generally unavailable as prey for syntonic bats (i.e., bats that use echolocation frequencies between 20 and 50 kHz within the hearing range of eared moths) due to the moths' ability to detect syntonic bat echolocation. Syntonic bats therefore feed mainly on beetles, flies, true bugs, and non-eared moths. The AFH is expected to be violated around lights where eared moths are susceptible to exploitation by syntonic bats because moths' evasive strategies become less effective. The hypothesis has been tested to date almost exclusively in areas with permanent lighting, where the effects of lights on bat diets are confounded with other aspects of human habitat alteration. We undertook diet analysis in areas with short-term, localized artificial lighting to isolate the effects of artificial lighting and determine if syntonic and allotonic bats (i.e., bats that use echolocation frequencies outside the hearing range of eared moths) consumed more moths under conditions of artificial lights than in natural darkness. We found that syntonic bats increased their consumption of moth prey under experimentally lit conditions, likely owing to a reduction in the ability of eared moths to evade the bats. Eared moths may increase in diets of generalist syntonic bats foraging around artificial light sources, as opposed to allotonic species and syntonic species with a more specialized diet.

Testing the "Fasting While Foraging" Hypothesis: Effects of Recent Feeding on Plasma Metabolite Concentrations in Little Brown Bats (Myotis lucifugus).

Plasma metabolite concentrations can be used to understand nutritional status and foraging behavior across ecological contexts including prehibernation fattening, migration refueling, and variation in foraging habitat quality. Generally, high plasma concentrations of the ketone ß-hydroxybutyrate, a product of fat catabolism, indicate fasting, while triglycerides indicate recent feeding and fat accumulation. In recent studies of insectivorous bats, triglyceride concentration increased after feeding as expected, but ß-hydroxybutyrate also unexpectedly increased rather than decreased. An aerial-hawking foraging strategy is energetically demanding, and thus it has been hypothesized that foraging by insectivorous bats requires catabolism of stored fat. We tested this hypothesis by quantifying plasma ß-hydroxybutyrate and triglyceride concentration following feeding in little brown bats (Myotis lucifugus) that were temporarily housed in individual cages to prevent flight. We provided a fixed amount of food and collected blood samples at different intervals after feeding to produce variation in plasma metabolite concentrations. Plasma triglyceride concentration responded as predicted, but similar to previous studies and contrary to our prediction, when flight was eliminated plasma ß-hydroxybutyrate concentration responded similarly to triglyceride. Thus, it is unlikely that the unexpected plasma ß-hydroxybutyrate patterns observed in previous studies were related to flight. The mechanism underlying this unexpected pattern remains unknown, but the response has been consistent in all studies to date. Thus, plasma metabolite analysis provides an effective tool for studies of nutritional status, although more work is needed to understand why insectivorous bats respond differently than other taxa.

Community Physiological Ecology.

The effects of animal homeostatic function on ecological interactions have not been well-integrated into community ecology. Animals mediate environmental change and stressors through homeostatic shifts in physiology and behavior, which likely shape ecological interactions and plant communities. Animal responses to stressors can alter their habitat use, selective foraging, and stoichiometry, which can in turn affect trophic interactions, plant growth, reproduction, and dispersal. Here, we describe a community physiological ecology framework that integrates classical ecological theory and emerging empirical approaches to test how animal homeostatic responses to environmental change mediate ecological interactions and shape communities. Interdisciplinary approaches could provide essential data to characterize and forecast community responses to rapid global environmental change.

Illuminating the physiological implications of artificial light on an insectivorous bat community.

Global light pollution threatens to disturb numerous wildlife species, but impacts of artificial light will likely vary among species within a community. Thus, artificial lights may change the environment in such a way as to create winners and losers as some species benefit while others do not. Insectivorous bats are nocturnal and a good model to test for differential effects of light pollution on a single community. We used a physiological technique to address this community-level question by measuring plasma ß-hydroxybutyrate (a blood metabolite) concentrations from six species of insectivorous bats in lit and unlit conditions. We also recorded bat calls acoustically to measure activity levels between experimental conditions. Blood metabolite level and acoustic activity data suggest species-specific changes in foraging around lights. In red bats (Lasiurus borealis), ß-hydroxybutyrate levels at lit sites were highest early in the night before decreasing. Acoustic data indicate pronounced peaks in activity at lit sites early in the night. In red bats on dark nights and in the other species in this community, which seem to avoid lights, ß-hydroxybutyrate remained relatively constant. Our results suggest red bats are more willing to expend energy to actively forage around lights despite potential negative impacts, while other, generally rarer species avoid lit areas. Artificial light appears to have a bifurcating effect on bat communities, whereby some species take advantage of concentrated prey resources, yet most do not. Further, this may concentrate light-intolerant species into limited dark refugia, thereby increasing competition for depauperate, phototactic insect communities.

Emerging threat of the 21st century lightscape to global biodiversity.

Over the last century the temporal and spatial distribution of light on Earth has been drastically altered by human activity. Despite mounting evidence of detrimental effects of light pollution on organisms and their trophic interactions, the extent to which light pollution threatens biodiversity on a global scale remains unclear. We assessed the spatial extent and magnitude of light encroachment by measuring change in the extent of light using satellite imagery from 1992 to 2012 relative to species richness for terrestrial and freshwater mammals, birds, reptiles, and amphibians. The encroachment of light into previously dark areas was consistently high, often doubling, in areas of high species richness for all four groups. This pattern persisted for nocturnal groups (e.g., bats, owls, and geckos) and species considered vulnerable to extinction. Areas with high species richness and large increases in light extent were clustered within newly industrialized regions where expansion of light is likely to continue unabated unless we act to conserve remaining darkness. Implementing change at a global scale requires global public, and therefore scientific, support. Here, we offer substantial evidence that light extent is increasing where biodiversity is high, representing an emerging threat to global biodiversity requiring immediate attention.

Exogenous stress hormones alter energetic and nutrient costs of development and metamorphosis.

Variation in environmental conditions during larval life stages can shape development during critical windows and have lasting effects on the adult organism. Changes in larval developmental rates in response to environmental conditions, for example, can trade off with growth to determine body size and condition at metamorphosis, which can affect adult survival and fecundity. However, it is unclear how use of energy and nutrients shape trade-offs across life-stage transitions because no studies have quantified these costs of larval development and metamorphosis. We used an experimental approach to manipulate physiological stress in larval amphibians, along with respirometry and 13C-breath testing to quantify the energetic and nutritional costs of development and metamorphosis. Central to larval developmental responses to environmental conditions is the hypothalamic-pituitary-adrenal/interrenal (HPA/I) axis, which regulates development, as well as energy homeostasis and stress responses across many taxa. Given these pleiotropic effects of HPA/I activity, manipulation of the HPA/I axis may provide insight into costs of metamorphosis. We measured the energetic and nutritional costs across the entire larval period and metamorphosis in a larval amphibian exposed to exogenous glucocorticoid (GC) hormones - the primary hormone secreted by the HPA/I axis. We measured metabolic rates and dry mass across larval ontogeny, and quantified lipid stores and nutrient oxidation via 13C-breath testing during metamorphosis, under control and GC-exposed conditions. Changes in dry mass match metamorphic states previously reported in the literature, but dynamics of metabolism were influenced by the transition from aquatic to terrestrial respiration. GC-treated larvae had lower dry mass, decreased fat stores and higher oxygen consumption during stages where controls were conserving energy. GC-treated larvae also oxidized greater amounts of 13C-labelled protein stores. These results provide evidence for a proximate cause of the physiological trade-off between larval growth and development, and provide insight into the energetic and nutrient costs that shape fitness trade-offs across life stages.

Torpor Patterns in Desert Hedgehogs (Paraechinus aethiopicus) Represent Another New Point along a Thermoregulatory Continuum.

Documenting variation in thermoregulatory patterns across phylogenetically and geographically diverse taxa is key to understanding the evolution of endothermy and heterothermy in birds and mammals. We recorded body temperature (Tb) in free-ranging desert hedgehogs (Paraechinus aethiopicus) across three seasons in the deserts of Saudi Arabia. Modal Tb's (35°-36.5°C) were slightly below normal for mammals but still warmer than those of other hedgehogs. The single maximum Tb recorded was 39.2°C, which is cooler than maximum Tb's recorded in most desert mammals. Desert hedgehogs commonly used torpor during winter and spring but never during summer. Torpor bouts occurred frequently but irregularly, and most lasted less than 24 h. Unlike daily heterotherms, desert hedgehogs did occasionally remain torpid for more than 24 h, including one bout of 101 h. Body temperatures during torpor were often within 2°-3°C of ambient temperature; however, we never recorded repeated bouts of long, predictable torpor punctuated by brief arousal periods similar to those common among seasonal hibernators. Thus, desert hedgehogs can be included on the ever-growing list of species that display torpor patterns intermediate to traditionally defined hibernators and daily heterotherms. Extant hedgehogs are a recent radiation within an ancient family, and the intermediate thermoregulatory pattern displayed by desert hedgehogs is unlike the deeper and more regular torpor seen in other hedgehogs, suggesting that this may be a derived-as opposed to ancestral-trait in this subfamily. We suggest that this family (Erinaceidae) and order (Eulipotyphla) may be important for understanding the evolution of thermoregulatory patterns among Laurasiatheria and mammals in general.

Physiological and behavioral adaptations in bats living at high latitudes.

Widespread animals at the extremes of the species' distribution experience ecological constraints different than individuals in the core of the distribution. For example, small endotherms at very high latitudes face short summers with cool temperatures and a lack of true darkness. In particular, insectivorous bats at high latitudes may experience constraints because of their unique life history traits, and may have different energy requirements than bats at lower latitudes. To evaluate the extent of these differences, we estimated an energy budget and refueling rates for reproductively active female little brown bats (Myotis lucifugus) roosting in buildings in eastern Alaska (~63°N). Physiological parameters (torpor use and metabolic rates) and daily energy expenditures (25.7±5.3kJd(-1)) were similar to, or slightly lower than, conspecifics at lower latitudes. Northern little brown bats foraged for less time than southerly conspecifics, but measurements of plasma ß-hydroxybutyrate concentrations suggest that northern bats refuel at a rate considerably higher than those to the south. It appears that high refueling rates (and therefore foraging intensity) involve a dietary shift to orb-weaver spiders, which are abundant and likely offer higher energetic benefit than the small, flying insects consumed by individuals in other parts of the distribution. Environmental factors may limit species' distributions, but our results provide an example of a population at the limit of their geographic range that has compensated for environmental challenges by adopting unique behavioral strategies while the underlying physiology (including daily energy expenditure) remains similar to populations at the core of the species' range.

Bats initiate vital agroecological interactions in corn.

In agroecosystems worldwide, bats are voracious predators of crop pests and may provide services to farmers worth billions of U.S. dollars. However, such valuations make untested assumptions about the ecological effect of bats in agroecosystems. Specifically, estimates of the value of pest suppression services assume bats consume sufficient numbers of crop pests to affect impact pest reproduction and subsequent damage to crops. Corn is an essential crop for farmers, and is grown on more than 150 million hectares worldwide. Using large exclosures in corn fields, we show that bats exert sufficient pressure on crop pests to suppress larval densities and damage in this cosmopolitan crop. In addition, we show that bats suppress pest-associated fungal growth and mycotoxin in corn. We estimate the suppression of herbivory by insectivorous bats is worth more than 1 billion USD globally on this crop alone, and bats may further benefit farmers by indirectly suppressing pest-associated fungal growth and toxic compounds on corn. Bats face a variety of threats globally, but their relevance as predators of insects in ubiquitous corn-dominated landscapes underlines the economic and ecological importance of conserving biodiversity.

Interruption to cutaneous gas exchange is not a likely mechanism of WNS-associated death in bats.

Pseudogymnoascus destructans is the causative fungal agent of white-nose syndrome (WNS), an emerging fungal-borne epizootic. WNS is responsible for a catastrophic decline of hibernating bats in North America, yet we have limited understanding of the physiological interactions between pathogen and host. Pseudogymnoascus destructans severely damages wings and tail membranes, by causing dryness that leads to whole sections crumbling off. Four possible mechanisms have been proposed by which infection could lead to dehydration; in this study, we tested one: P. destructans infection could cause disruption to passive gas-exchange pathways across the wing membranes, thereby causing a compensatory increase in water-intensive pulmonary respiration. We hypothesized that total evaporative water loss would be greater when passive gas exchange was inhibited. We found that bats did not lose more water when passive pathways were blocked. This study provides evidence against the proposed proximal mechanism that disruption to passive gas exchange causes dehydration and death to WNS-infected bats.