Non-Torpid Heterothermy in Mammals: Another Category along the Homeothermy-Hibernation Continuum.

Variability in body temperature is now recognized to be widespread among whole-body endotherms with homeothermy being the exception rather than the norm. A wide range of body temperature patterns exists in extant endotherms, spanning from strict homeothermy, to occasional use of torpor, to deep seasonal hibernation with many points in between. What is often lost in discussions of heterothermy in endotherms are the benefits of variations in body temperature outside of torpor. Endotherms that do not use torpor can still obtain extensive energy and water savings from varying levels of flexibility in normothermic body temperature regulation. Flexibility at higher temperatures (heat storage or facultative hyperthermia) can provide significant water savings, while decreases at cooler temperatures, even outside of torpor, can lower the energetic costs of thermoregulation during rest. We discuss the varying uses of the terms heterothermy, thermolability, and torpor to describe differences in the amplitude of body temperature cycles and advocate for a broader use of the term "heterothermy" to include non-torpid variations in body temperature.

Daily Torpor in Birds and Mammals: Past, Present, and Future of the Field.

Torpor is an incredibly efficient energy-saving strategy that many endothermic birds and mammals use to save energy, by lowering their metabolic rates, heart rates, and typically body temperatures. Over the last few decades, the study of daily torpor-in which torpor is used for less than 24 hours per bout-has advanced rapidly. The papers in this issue cover the ecological and evolutionary drivers of torpor, as well as some of the mechanisms governing torpor use. We identified broad focus areas that need special attention: clearly defining the various parameters that indicate torpor use and identifying the genetic and neurological mechanisms regulating torpor. Recent studies on daily torpor and heterothermy, including the ones in this issue, have furthered the field immensely. We look forward to a period of immense growth in this field.

Rare and Opportunistic Use of Torpor in Mammals-An Echo from the Past?

Torpor was traditionally seen as a winter survival mechanism employed by animals living in cold and highly seasonal habitats. Although we now know that torpor is also used by tropical and subtropical species, and in response to a variety of triggers, torpor is still largely viewed as a highly controlled, seasonal mechanism shown by Northern hemisphere species. To scrutinize this view, we report data from a macroanalysis in which we characterized the type and seasonality of torpor use from mammal species currently known to use torpor. Our findings suggest that predictable, seasonal torpor patterns reported for Northern temperate and polar species are highly derived forms of torpor expression, whereas the more opportunistic and variable forms of torpor that we see in tropical and subtropical species are likely closer to the patterns expressed by ancestral mammals. Our data emphasize that the torpor patterns observed in the tropics and subtropics should be considered the norm and not the exception.

Microclimate conditions alter Ixodes scapularis (Acari: Ixodidae) overwinter survival across climate gradients in Maine, United States.

The incidence and geographic range of vector-borne diseases have been expanding in recent decades, attributed in part to global climate change. Blacklegged ticks (Ixodes scapularis), the primary vector for multiple tick-borne pathogens in North America, are spreading rapidly beyond their historic post-colonial range and are thought to be constrained mainly by winter temperature at northern latitudes. Our research explored whether winter climate currently limits the distribution of blacklegged ticks and the pathogens they transmit in Maine, U.S.A., by contributing to overwinter mortality of nymphs. We experimentally tested tick overwinter survival across large-scale temperature and snowfall gradients and assessed factors contributing to winter mortality in locations where blacklegged tick populations are currently established and locations where the blacklegged tick has not yet been detected. We also tested the hypothesis that insulation in the tick microhabitat (i.e., by leaf litter and snowpack) can facilitate winter survival of blacklegged tick nymphs despite inhospitable ambient conditions. Overwinter survival was not significantly different in coastal southern compared to coastal and inland northern Maine, most likely due to sufficient snowpack that protected against low ambient temperatures at high latitudes. Snow cover and leaf litter contributed significantly to overwinter survival at sites in both southern and northern Maine. To further assess whether the current distribution of blacklegged ticks in Maine aligns with patterns of overwinter survival, we systematically searched for and collected ticks at seven sites along latitudinal and coastal-inland climate gradients across the state. We found higher densities of blacklegged ticks in coastal southern Maine (90.2 ticks/1000 m2) than inland central Maine (17.8 ticks/1000 m2) and no blacklegged ticks in inland northern Maine. Our results suggest that overwinter survival is not the sole constraint on the blacklegged tick distribution even under extremely cold ambient conditions and additional mechanisms may limit the continued northward expansion of ticks.

Understanding Drivers of Variation and Predicting Variability Across Levels of Biological Organization.

Differences within a biological system are ubiquitous, creating variation in nature. Variation underlies all evolutionary processes and allows persistence and resilience in changing environments; thus, uncovering the drivers of variation is critical. The growing recognition that variation is central to biology presents a timely opportunity for determining unifying principles that drive variation across biological levels of organization. Currently, most studies that consider variation are focused at a single biological level and not integrated into a broader perspective. Here we explain what variation is and how it can be measured. We then discuss the importance of variation in natural systems, and briefly describe the biological research that has focused on variation. We outline some of the barriers and solutions to studying variation and its drivers in biological systems. Finally, we detail the challenges and opportunities that may arise when studying the drivers of variation due to the multi-level nature of biological systems. Examining the drivers of variation will lead to a reintegration of biology. It will further forge interdisciplinary collaborations and open opportunities for training diverse quantitative biologists. We anticipate that these insights will inspire new questions and new analytic tools to study the fundamental questions of what drives variation in biological systems and how variation has shaped life.

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.

Do endotherms have thermal performance curves?

Temperature is an important environmental factor governing the ability of organisms to grow, survive and reproduce. Thermal performance curves (TPCs), with some caveats, are useful for charting the relationship between body temperature and some measure of performance in ectotherms, and provide a standardized set of characteristics for interspecific comparisons. Endotherms, however, have a more complicated relationship with environmental temperature, as endothermy leads to a decoupling of body temperature from external temperature through use of metabolic heat production, large changes in insulation and variable rates of evaporative heat loss. This has impeded our ability to model endothermic performance in relation to environmental temperature as well as to readily compare performance between species. In this Commentary, we compare the strengths and weaknesses of potential TPC analogues (including other useful proxies for linking performance to temperature) in endotherms and suggest several ways forward in the comparative ecophysiology of endotherms. Our goal is to provide a common language with which ecologists and physiologists can evaluate the effects of temperature on performance. Key directions for improving our understanding of endotherm thermoregulatory physiology include a comparative approach to the study of the level and precision of body temperature, measuring performance directly over a range of body temperatures and building comprehensive mechanistic models of endotherm responses to environmental temperatures. We believe the answer to the question posed in the title could be 'yes', but only if 'performance' is well defined and understood in relation to body temperature variation, and the costs and benefits of endothermy are specifically modelled.

Activity analysis of thermal imaging videos using a difference imaging approach.

Infrared thermal imaging is a passive imaging technique that captures the emitted radiation from an object to estimate surface temperature, often for inference of heat transfer. Infrared thermal imaging offers the potential to detect movement without the challenges of glare, shadows, or changes in lighting associated with visual digital imaging or active infrared imaging. In this paper, we employ a frame subtraction algorithm for extracting the pixel-by-pixel relative change in signal from a fixed focus video file, tailored for use with thermal imaging videos. By summing the absolute differences across an entire video, we are able to assign quantitative activity assessments to thermal imaging data for comparison with simultaneous recordings of metabolic rates. We tested the accuracy and limits of this approach by analyzing movement of a metronome and provide an example application of the approach to a study of Darwin's finches. In principle, this "Difference Imaging Thermography" (DIT) would allow for activity data to be standardized to energetic measurements and could be applied to any radiometric imaging system.

Ambient Temperature Cycles Affect Daily Torpor and Hibernation Patterns in Malagasy Tenrecs.

Hibernation and daily torpor (heterothermy) allow endotherms to cope with demanding environmental conditions. The depth and duration of torpor bouts vary considerably between tropical and temperate climates, and tropical hibernators manage to cope with a wider spectrum of ambient temperature (T a) regimes during heterothermy. As cycles in T a can have profound effects on activity and torpor patterns as well as energy expenditure, we examined how these characteristics are affected by daily fluctuating versus constant T a in a tropical hibernator, the lesser hedgehog tenrec (Echinops telfairi). Throughout the study, regardless of season, the tenrecs became torpid every day. In summer, E. telfairi used daily fluctuations in T a to passively rewarm from daily torpor, which led to synchrony in the activity phases and torpor bouts between individuals and generally decreased energy expenditure. In contrast, animals housed at constant T a showed considerable variation in timing and they had to invest more energy through endogenous heat production. During the hibernation season (winter) E. telfairi hibernated for several months in constant, as well as in fluctuating T a and, as in summer, under fluctuating T a arousals were much more uniform and showed less variation in timing compared to constant temperature regimes. The timing of torpor is not only important for its effective use, but synchronization of activity patterns could also be essential for social interactions, and successful foraging bouts. Our results highlight that T a cycles can be an effective zeitgeber for activity and thermoregulatory rhythms throughout the year and that consideration should be given to the choice of temperature regime when studying heterothermy under laboratory conditions.

Small Tropical Mammals Can Take the Heat: High Upper Limits of Thermoneutrality in a Bornean Treeshrew.

Tropical ectotherms are generally believed to be more vulnerable to global heating than temperate species. Currently, however, we have insufficient knowledge of the thermoregulatory physiology of equatorial tropical mammals, particularly of small diurnal mammals, to enable similar predictions. In this study, we measured the resting metabolic rates (via oxygen consumption) of wild-caught lesser treeshrews (Tupaia minor, order Scandentia) over a range of ambient temperatures. We predicted that, similar to other treeshrews, T. minor would exhibit more flexibility in body temperature regulation and a wider thermoneutral zone compared with other small mammals because these thermoregulatory traits provide both energy and water savings at high ambient temperatures. Basal metabolic rate was on average 1.03±0.10 mL O2 h-1 g-1, which is within the range predicted for a 65-g mammal. We calculated the lower critical temperature of the thermoneutral zone at 31.0°C (95% confidence interval: 29.3°-32.7°C), but using metabolic rates alone, we could not determine the upper critical temperature at ambient temperatures as high as 36°C. The thermoregulatory characteristics of lesser treeshrews provide a means of saving energy and water at temperatures well in excess of their current environmental temperatures. Our research highlights the knowledge gaps in our understanding of the energetics of mammals living in high-temperature environments, specifically in the equatorial tropics, and questions the purported lack of variance in the upper critical temperatures of the thermoneutral zone in mammals, emphasizing the importance of further research in the tropics.

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.

Staying hot to fight the heat-high body temperatures accompany a diurnal endothermic lifestyle in the tropics.

Much of our knowledge of the thermoregulation of endotherms has been obtained from species inhabiting cold and temperate climates, our knowledge of the thermoregulatory physiology of tropical endotherms is scarce. We studied the thermoregulatory physiology of a small, tropical mammal, the large treeshrew (Tupaia tana, Order Scandentia) by recording the body temperatures of free-ranging individuals, and by measuring the resting metabolic rates of wild individuals held temporarily in captivity. The amplitude of daily body temperature (~ 4 °C) was higher in treeshrews than in many homeothermic eutherian mammals; a consequence of high active-phase body temperatures (~ 40 °C), and relatively low rest-phase body temperatures (~ 36 °C). We hypothesized that high body temperatures enable T. tana to maintain a suitable gradient between ambient and body temperature to allow for passive heat dissipation, important in high-humidity environments where opportunities for evaporative cooling are rare. Whether this thermoregulatory phenotype is unique to Scandentians, or whether other warm-climate diurnal small mammals share similar thermoregulatory characteristics, is currently unknown.

Embracing heterothermic diversity: non-stationary waveform analysis of temperature variation in endotherms.

Recent research is revealing incredible diversity in the thermoregulatory patterns of wild and captive endotherms. As a result of these findings, classic thermoregulatory categories of 'homeothermy', 'daily heterothermy', and 'hibernation' are becoming harder to delineate, impeding our understanding of the physiological and evolutionary significance of variation within and around these categories. However, we lack a generalized analytical approach for evaluating and comparing the complex and diversified nature of the full breadth of heterothermy expressed by individuals, populations, and species. Here we propose a new approach that decomposes body temperature time series into three inherent properties-waveform, amplitude, and period-using a non-stationary technique that accommodates the temporal variability of body temperature patterns. This approach quantifies circadian and seasonal variation in thermoregulatory patterns, and uses the distribution of observed thermoregulatory patterns as a basis for intra- and inter-specific comparisons. We analyse body temperature time series from multiple species, including classical hibernators, tropical heterotherms, and homeotherms, to highlight the approach's general usefulness and the major axes of thermoregulatory variation that it reveals.

Can we predict ectotherm responses to climate change using thermal performance curves and body temperatures?

Thermal performance curves (TPCs), which quantify how an ectotherm's body temperature (Tb ) affects its performance or fitness, are often used in an attempt to predict organismal responses to climate change. Here, we examine the key - but often biologically unreasonable - assumptions underlying this approach; for example, that physiology and thermal regimes are invariant over ontogeny, space and time, and also that TPCs are independent of previously experienced Tb. We show how a critical consideration of these assumptions can lead to biologically useful hypotheses and experimental designs. For example, rather than assuming that TPCs are fixed during ontogeny, one can measure TPCs for each major life stage and incorporate these into stage-specific ecological models to reveal the life stage most likely to be vulnerable to climate change. Our overall goal is to explicitly examine the assumptions underlying the integration of TPCs with Tb , to develop a framework within which empiricists can place their work within these limitations, and to facilitate the application of thermal physiology to understanding the biological implications of climate change.

Mammal survival at the Cretaceous-Palaeogene boundary: metabolic homeostasis in prolonged tropical hibernation in tenrecs.

Free-ranging common tenrecs, Tenrec ecaudatus, from sub-tropical Madagascar, displayed long-term (nine months) hibernation which lacked any evidence of periodic interbout arousals (IBAs). IBAs are the dominant feature of the mammalian hibernation phenotype and are thought to periodically restore long-term ischaemia damage and/or metabolic imbalances (depletions and accumulations). However, the lack of IBAs in tenrecs suggests no such pathology at hibernation Tbs > 22°C. The long period of tropical hibernation that we report might explain how the ancestral placental mammal survived the global devastation that drove the dinosaurs and many other vertebrates to extinction at the Cretaceous-Palaeogene boundary following a meteorite impact. The genetics and biochemistry of IBAs are of immense interest to biomedical researchers and space exploration scientists, in the latter case, those envisioning a hibernating state in astronauts for deep space travel. Unravelling the physiological thresholds and temperature dependence of IBAs will provide new impetus to these research quests.

Effects of reproductive status and high ambient temperatures on the body temperature of a free-ranging basoendotherm.

Tenrecs (Order Afrosoricida) exhibit some of the lowest body temperatures (T b) of any eutherian mammal. They also have a high level of variability in both active and resting T bs and, at least in cool temperatures in captivity, frequently employ both short- and long-term torpor. The use of heterothermy by captive animals is, however, generally reduced during gestation and lactation. We present data long-term T b recordings collected from free-ranging S. setosus over the course of two reproductive seasons. In general, reproductive females had slightly higher (~32 °C) and less variable T b, whereas non-reproductive females and males showed both a higher propensity for torpor as well as lower (~30.5 °C) and more variable rest-phase T bs. Torpor expression defined using traditional means (using a threshold or cut-off T b) was much lower than predicted based on the high degree of heterothermy in captive tenrecs. However, torpor defined in this manner is likely to be underestimated in habitats where ambient temperature is close to T b. Our results caution against inferring metabolic states from T b alone and lend support to the recent call to define torpor in free-ranging animals based on mechanistic and not descriptive variables. In addition, lower variability in T b observed during gestation and lactation confirms that homeothermy is essential for reproduction in this species and probably for basoendothermic mammals in general. The relatively low costs of maintaining homeothermy in a sub-tropical environment might help shed light on how homeothermy could have evolved incrementally from an ancestral heterothermic condition.

Increased homeothermy during reproduction in a basal placental mammal.

Homeothermic endothermy, the maintenance of a high and stable body temperature (Tb) using heat produced by elevated metabolism, is energetically expensive. There is increasing evidence that the earliest endotherms were heterotherms that, rather than maintaining strict homeothermy, allowed Tb to fluctuate with large variations between active and rest-phase Tb. The high level of homeothermy observed in modern mammals is therefore likely to have evolved from an ancestral heterothermic state. One of the hypotheses for the evolution of endothermy is that homeothermy allows for greater energetic output during reproduction (parental care model). We tested this hypothesis by measuring metabolic rates over a range of ambient temperatures in both reproductive and non-reproductive greater hedgehog tenrecs (Setifer setosus), a physiologically primitive mammal from Madagascar. Tenrecs have some of the lowest metabolic rates and highest levels of Tb variability of any mammal and are therefore good models of the ancestral eutherian state. During pregnancy and lactation, there was an increase in metabolism and Tb below the thermoneutral zone, accompanied by a decrease in Tb variability. The lower critical limit of the thermoneutral zone was estimated at ~25°C. However, whereas increases in resting metabolism were substantial below 20°C (up to 150% higher during reproduction), daytime rest-phase ambient temperatures at the study site rarely reached equivalent low levels. Thus, S. setosus provide an example for how relatively low-cost increases in homeothermy could have led to substantial increases in fitness by allowing for the faster production of young. The mechanisms necessary for increases in thermogenesis during reproduction would have further benefited the development of homeothermy in mammals.

Seasonal torpor and normothermic energy metabolism in the Eastern chipmunk (Tamias striatus).

To assess the changes in thermoregulatory characteristics that accompany the seasonal expression of torpor we measured seasonal differences in body mass adjustments, body temperature (T (b)) and metabolic rate (MR) in both summer- and winter-acclimated individuals from a species of food-storing hibernator, the Eastern chipmunk (Tamias striatus). Torpor occurred only in the winter and was associated with lower normothermic T (b), during inter-bout arousal periods than in the summer. Chipmunks increased body mass before the initiation of torpor in winter, and steadily lost mass as the hibernation season progressed. Torpor expression was correlated to initial mass gain, with the individuals who showed the largest mass increase in the fall showing the highest degree of torpor. Acclimation to winter-like conditions produced a decline in normothermic MR at all ambient temperatures examined. The findings indicate that torpor expression is accompanied by a decrease in T (b) and MR during normothermy, indicating that a conservation of energy metabolism occurs, not only in torpor, but also during the inter-bout arousal periods.

Seasonal changes in thermoregulatory responses to hypoxia in the Eastern chipmunk (Tamias striatus).

Mammalian heterotherms are known to be more tolerant of low oxygen levels than homeotherms. However, heterotherms demonstrate extreme seasonality in daily heterothermy and torpor expression. Because hypoxia depresses body temperature (T(b)) and metabolism in mammals, it was of interest to see if seasonal comparisons of normothermic animals of a species capable of hibernation produce changes in their responses to hypoxia that would reflect a seasonal change in hypoxia tolerance. The species studied, the Eastern chipmunk (Tamias striatus, Linnaeus 1758), is known to enter into torpor exclusively in the winter. To test for seasonal differences in the metabolic and thermoregulatory responses to hypoxia (9.9 kPa), flow-through respirometry was used to compare oxygen consumption, minimum thermal conductance and T(b) under fixed ambient temperature (T(a)) conditions whereas a thermal gradient was used to assess selected T(a) and T(b) in response to hypoxia, in both summer- and winter-acclimated animals. No differences were observed between seasons in resting metabolism or thermal conductance in normoxic, normothermic animals. Providing the animals with a choice of T(a) in hypoxia attenuated the hypoxic drop in T(b) in both seasons, suggesting that the reported fall in T(b) in hypoxia is not fully manifested in the behavioural pathways responsible for thermoregulation in chipmunks. Instead, T(b) in hypoxia tends to be more variable and dependent on both T(a) and season. Although T(b) dropped in hypoxia in both seasons, the decrease was less in the winter with no corresponding decrease in metabolism, indicating that winter chipmunks are more tolerant to hypoxia than summer animals.