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.

Saving energy via short and shallow torpor bouts.

Maintaining a high and stable body temperature as observed in most endothermic mammals and birds is energetically costly and many heterothermic species reduce their metabolic demands during energetic bottlenecks through the use of torpor. With the increasing number of heterotherms revealed in a diversity of habitats, it becomes apparent that triggers and patterns of torpor use are more variable than previously thought. Here, we report the previously overlooked use of, shallow rest-time torpor (body temperature >30 °C) in African lesser bushbabies, Galago moholi. Body core temperature of three adult male bushbabies recorded over five months showed a clear bimodal distribution with an average active modal temperature of 39.2 °C and a resting modal body temperature of 36.7 °C. Shallow torpor was observed in two out of three males (n = 29 torpor bouts) between June and August (austral winter), with body temperatures dropping to an overall minimum of 30.7 °C and calculated energy savings of up to 10%. We suggest that shallow torpor may be an ecologically important, yet mostly overlooked energy-saving strategy employed by heterothermic mammals. Our data emphasise that torpor threshold temperatures need to be used with care if we aim to fully understand the level of physiological plasticity displayed by heterothermic species.

Food availability positively affects the survival and somatic maintenance of hibernating garden dormice (Eliomys quercinus).

BACKGROUND: Torpor is an energy saving strategy achieved by substantial reductions of metabolic rate and body temperature that enables animals to survive periods of low resource availability. During hibernation (multiday torpor), the frequency of periodic rewarming-characterised by high levels of oxidative stress-is associated with shortening of telomeres, a marker of somatic maintenance. OBJECTIVES: In this study, we determined the impact of ambient temperature on feeding behaviour and telomere dynamics in hibernating garden dormice (Eliomys quercinus) over winter. This obligate hibernator prepares for hibernation by accumulating fat stores but can also feed during hibernation. METHODOLOGY: Food intake, torpor pattern, changes in telomere length, and body mass change were assessed in animals housed at experimentally controlled temperatures of either 14 °C (i.e., a mild winter) or 3 °C (i.e., a cold winter) over 6 months. RESULTS: When hibernating at 14 °C, dormice experienced 1.7-fold more frequent and 2.4-fold longer inter-bout euthermia, and spent significantly less time torpid, compared to animals hibernating at 3 °C. Higher food intake enabled individuals to compensate for increased energetic costs when hibernating at milder temperatures (14 °C vs. 3 °C), to buffer body mass loss and thus increase winter survival. Interestingly, we observed a significant increase of telomere length over the entire hibernation period, irrespective of temperature treatment. CONCLUSION: We conclude that higher temperatures during winter, if associated with sufficient food availability, can have a positive effect on the individual's energy balance and somatic maintenance. These results suggest that winter food availability might be a crucial determinant for the survival of the garden dormouse in the context of ever-increasing environmental temperatures.

Whole-body endothermy: ancient, homologous and widespread among the ancestors of mammals, birds and crocodylians.

The whole-body (tachymetabolic) endothermy seen in modern birds and mammals is long held to have evolved independently in each group, a reasonable assumption when it was believed that its earliest appearances in birds and mammals arose many millions of years apart. That assumption is consistent with current acceptance that the non-shivering thermogenesis (NST) component of regulatory body heat originates differently in each group: from skeletal muscle in birds and from brown adipose tissue (BAT) in mammals. However, BAT is absent in monotremes, marsupials, and many eutherians, all whole-body endotherms. Indeed, recent research implies that BAT-driven NST originated more recently and that the biochemical processes driving muscle NST in birds, many modern mammals and the ancestors of both may be similar, deriving from controlled 'slippage' of Ca2+ from the sarcoplasmic reticulum Ca2+ -ATPase (SERCA) in skeletal muscle, similar to a process seen in some fishes. This similarity prompted our realisation that the capacity for whole-body endothermy could even have pre-dated the divergence of Amniota into Synapsida and Sauropsida, leading us to hypothesise the homology of whole-body endothermy in birds and mammals, in contrast to the current assumption of their independent (convergent) evolution. To explore the extent of similarity between muscle NST in mammals and birds we undertook a detailed review of these processes and their control in each group. We found considerable but not complete similarity between them: in extant mammals the 'slippage' is controlled by the protein sarcolipin (SLN), in birds the SLN is slightly different structurally and its role in NST is not yet proved. However, considering the multi-millions of years since the separation of synapsids and diapsids, we consider that the similarity between NST production in birds and mammals is consistent with their whole-body endothermy being homologous. If so, we should expect to find evidence for it much earlier and more widespread among extinct amniotes than is currently recognised. Accordingly, we conducted an extensive survey of the palaeontological literature using established proxies. Fossil bone histology reveals evidence of sustained rapid growth rates indicating tachymetabolism. Large body size and erect stature indicate high systemic arterial blood pressures and four-chambered hearts, characteristic of tachymetabolism. Large nutrient foramina in long bones are indicative of high bone perfusion for rapid somatic growth and for repair of microfractures caused by intense locomotion. Obligate bipedality appeared early and only in whole-body endotherms. Isotopic profiles of fossil material indicate endothermic levels of body temperature. These proxies led us to compelling evidence for the widespread occurrence of whole-body endothermy among numerous extinct synapsids and sauropsids, and very early in each clade's family tree. These results are consistent with and support our hypothesis that tachymetabolic endothermy is plesiomorphic in Amniota. A hypothetical structure for the heart of the earliest endothermic amniotes is proposed. We conclude that there is strong evidence for whole-body endothermy being ancient and widespread among amniotes and that the similarity of biochemical processes driving muscle NST in extant birds and mammals strengthens the case for its plesiomorphy.

Survivable hypothermia or torpor in a wild-living rat: rare insights broaden our understanding of endothermic physiology.

Maintaining a high and stable body temperature as observed in endothermic mammals and birds is energetically costly. Thus, it is not surprising that we discover more and more heterothermic species that can reduce their energetic needs during energetic bottlenecks through the use of torpor. However, not all heterothermic animals use torpor on a regular basis. Torpor may also be important to an individual's probability of survival, and hence fitness, when used infrequently. We here report the observation of a single, ~ 5.5 h long hypothermic bout with a decrease in body temperature by 12 °C in the native Australian bush rat (Rattus fuscipes). Our data suggest that bush rats are able to rewarm from a body temperature of 24 °C, albeit with a rewarming rate lower than that expected on the basis of their body mass. Heterothermy, i.e. the ability to withstand and overcome periods of reduced body temperature, is assumed to be an evolutionarily ancestral (plesiomorphic) trait. We thus argue that such rare hypothermic events in species that otherwise appear to be strictly homeothermic could be heterothermic rudiments, i.e. a less derived form of torpor with limited capacity for rewarming. Importantly, observations of rare and extreme thermoregulatory responses by wild animals are more likely to be discovered with long-term data sets and may not only provide valuable insight about the physiological capability of a population, but can also help us to understand the constraints and evolutionary pathways of different phenologies.

The tarnished silver spoon? Trade-off between prenatal growth and telomere length in wild boar.

Life-history theory predicts a trade-off between growth rates and lifespan, which is reflected by telomere length, a biomarker of somatic state. We investigated the correlation between telomere length and early-life growth of wild boar piglets, Sus scrofa, kept under semi-natural conditions with high food availability to examine our hypothesis that increased pre- and postnatal growth will lead to telomere length attrition, but that a high supply of nutrient may provide the possibility to compensate telomere loss via telomere repair mechanisms. As predicted, our data showed a clear negative correlation between birth body mass and initial telomere length: heavier neonates had shorter telomeres at birth, and we did not find an influence of the mother on initial telomere length. Body mass at birth correlated with body mass later in life and postnatal growth rate did not affect telomere length. We observed an increase in telomere length during postnatal development, suggesting that high food availability allowed piglets to invest into both, growth and telomere restoration. The increase in telomere length over the duration of the study was not accompanied by telomerase activity; thus, telomere elongation was caused either by alternative mechanisms or by short pulses of telomerase activity that we missed. Taken together, this study suggests a trade-off between investment into growth and telomere maintenance even before birth and the possibility to compensate telomere attrition during growth under high amounts of available energy.

Cardiovascular effects of intravenous vatinoxan in wild boars (Sus scrofa) anaesthetised with intramuscular medetomidine-tiletamine-zolazepam.

BACKGROUND: The potent sedative medetomidine is a commonly used adjunct for the immobilisation of non-domestic mammals. However, its use is associated with pronounced cardiovascular side effects, such as bradycardia, vasoconstriction and decreased cardiac output. We investigated the effects of the peripherally-acting alpha-2-adrenoceptor antagonist vatinoxan on cardiovascular properties in medetomidine-tiletamine-zolazepam anaesthetised wild boar (Sus scrofa). METHODS: Twelve wild boars, anaesthetised twice with medetomidine (0.1 mg/kg) and tiletamine/zolazepam (2.5 mg/kg) IM in a randomised, crossover study, were administered (0.1 mg/kg) vatinoxan or an equivalent volume of saline IV (control). Cardiovascular variables, including heart rate (HR), mean arterial blood pressure (MAP), pulmonary artery pressure (PAP), pulmonary artery occlusion pressure (PAOP) and cardiac output (CO), were assessed 5 min prior to vatinoxan/saline administration until the end of anaesthesia 30 min later. RESULTS: MAP (p < 0.0001), MPAP (p < 0.001) and MPAOP (p < 0.0001) significantly decreased from baseline after vatinoxan until the end of anaesthesia. HR increased significantly (p < 0.0001) from baseline after vatinoxan administration. However, the effect on HR subsided 3 min after vatinoxan. All variables remained constant after saline injection. There was no significant effect of vatinoxan or saline on CO. CONCLUSION: Vatinoxan significantly reduced systemic and pulmonary artery hypertension, induced by medetomidine in wild boar.

Open-flow respirometry under field conditions: How does the airflow through the nest influence our results?

Open-flow respirometry is a common method to measure oxygen-uptake as a proxy of energy expenditure of organisms in real-time. Although most often used in the laboratory it has seen increasing application under field conditions. Air is drawn or pushed through a metabolic chamber or the nest with the animal, and the O2 depletion and/or CO2 accumulation in the air is analysed to calculate metabolic rate and energy expenditure. Under field conditions, animals are often measured within the microclimate of their nest and in contrast to laboratory work, the temperature of the air entering the nest cannot be controlled. Thus, the aim of our study was to determine the explanatory power of respirometry in a set-up mimicking field conditions. We measured O2 consumption of 14 laboratory mice (Mus musculus) using three different flow rates [50 L*h-1 (834 mL*min-1), 60 L*h-1 (1000 mL*min-1) and 70 L*h-1 (1167 mL*min-1)] and two different temperatures of the inflowing air; either the same as the temperature inside the metabolic chamber (no temperature differential; 20 °C), or cooler (temperature differential of 10 °C). Our results show that the energy expenditure of the mice did not change significantly in relation to a cooler airflow, nor was it affected by different flow rates, despite a slight, but significant decrease of about 1.5 °C in chamber temperature with the cooler airflow. Our study emphasises the validity of the results obtained by open-flow respirometry when investigating energy budgets and physiological responses of animals to ambient conditions. Nevertheless, subtle changes in chamber temperature in response to changes in the temperature and flow rate of the air pulled or pushed through the system were detectable. Thus, constant airflow during open-flow respirometry and consequent changes in nest/chamber temperature should be measured.

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.

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.

Always a price to pay: hibernation at low temperatures comes with a trade-off between energy savings and telomere damage.

We experimentally tested the costs of deep torpor at low temperatures by comparing telomere dynamics in two species of rodents hibernating at either 3 or 14°C. Our data show that hibernators kept at the warmer temperature had higher arousal frequencies, but maintained longer telomeres than individuals hibernating at the colder temperature. We suggest that the high-energy demand of frequent arousals is counteracted by a lower temperature differential between torpid and euthermic body temperature and that telomere length is restored during arousals when the body temperature is returned to normothermic values. Taken together, our study shows that hibernation at low body temperatures comes with costs on a cellular level and that hibernators need to actively counterbalance the shortening of telomeres.

Muscle nonshivering thermogenesis in a feral mammal.

Muscle nonshivering thermogenesis (NST) was recently suggested to play an important role in thermoregulation of species lacking brown adipose tissue (BAT). The mechanism, which is independent of muscle contractions, produces heat based on the activity of an ATPase pump in the sarcoplasmic reticulum (SERCA1a) and is controlled by the protein sarcolipin. To evaluate whether muscle NST could indeed play an important role in thermoregulation in species lacking BAT, we investigated the thermogenic capacities of newborn wild boar piglets. During cold exposure over the first 5 days of life, total heat production was improved while shivering intensity decreased, indicating an increasing contribution of NST. Sampling skeletal muscle tissue for analyses of SERCA activity as well as gene expression of SERCA1a and sarcolipin, we found an age-related increase in all three variables as well as in body temperature. Hence, the improved thermogenesis during the development of wild boars is not due to shivering but explained by the observed increase in SERCA activity. Our results suggest that muscle NST may be the primary mechanism of heat production during cold stress in large mammals lacking BAT, strengthening the hypothesis that muscle NST has likely played an important role in the evolution of endothermy.

A burning question: what are the risks and benefits of mammalian torpor during and after fires?

Although wildfires are increasing globally, available information on how mammals respond behaviourally and physiologically to fires is scant. Despite a large number of ecological studies, often examining animal diversity and abundance before and after fires, the reasons as to why some species perform better than others remain obscure. We examine how especially small mammals, which generally have high rates of energy expenditure and food requirements, deal with fires and post-fire conditions. We evaluate whether mammalian torpor, characterised by substantial reductions in body temperature, metabolic rate and water loss, plays a functional role in survival of mammals impacted by fires. Importantly, torpor permits small mammals to reduce their activity and foraging, and to survive on limited food. Torpid small mammals (marsupials and bats) can respond to smoke and arouse from torpor, which provides them with the possibility to evade direct exposure to fire, although their response is often slowed when ambient temperature is low. Post-fire conditions increase expression of torpor with a concomitant decrease in activity for free-ranging echidnas and small forest-dwelling marsupials, in response to reduced cover and reduced availability of terrestrial insects. Presence of charcoal and ash increases torpor use by captive small marsupials beyond food restriction alone, likely in anticipation of detrimental post-fire conditions. Interestingly, although volant bats use torpor on every day after fires, they respond by decreasing torpor duration, and increasing activity, perhaps because of the decrease in clutter and increase in foraging opportunities due to an increase in aerial insects. Our summary shows that torpor is an important tool for post-fire survival and, although the physiological and behavioural responses of small mammals to fire are complex, they seem to reflect energetic requirements and mode of foraging. We make recommendations on the conditions during management burns that are least likely to impact heterothermic mammals.

An in Vitro Biosynthesis of Sesquiterpenes Starting from Acetic Acid.

The enzymatic synthesis of terpenes was investigated by using a cascade based on the mevalonic acid pathway. Suitable enzymes from all kingdoms of life were identified and combined to give rise to geosmin and patchoulol as representative compounds. The pathway was studied in three separate segments, which were subsequently combined in a ten-step cascade plus added cofactor regeneration systems. The cascade delivers farnesyl pyrophosphate with >40 % conversion and cyclises it to sesquiterpenes with >90 % conversion.

Physiological and behavioral responses of an arboreal mammal to smoke and charcoal-ash substrate.

The recent observation that torpor plays a key role in post-fire survival has been mainly attributed to the reduced food resources after fires. However, some of these adjustments can be facilitated or amplified by environmental changes associated with fires, such as the presence of a charcoal-ash substrate. In a previous experiment on a small terrestrial mammal the presence of charcoal and ash linked to food restriction intensified torpor use. However, whether fire cues also act as a trigger of torpor use when food is available and whether they affect other species including arboreal mammals remains elusive. To evaluate whether smoke, charcoal and ash can act as proximate triggers for an impending period of food shortage requiring torpor for mammals, we conducted an experiment on captive sugar gliders (Petaurus breviceps), a small, arboreal marsupial, housed in outside aviaries under different food regimes and natural ambient conditions. When food was available, fire simulation via exposure to smoke and charcoal-ash substrate caused a significant earlier start of activity and a significant decrease in resting body temperature. In contrast, only when food was withheld, did smoke and charcoal-ash exposure significantly enhance torpor depth and duration. Thus, our study not only provides evidence that fire simulation does affect arboreal and terrestrial species similarly, but also suggests that smoke and ash were presumably selected as cues for torpor induction because they indicate an impending lack of food.

A Multiperspective Approach to Solvent Regulation of Enzymatic Activity: HMG-CoA Reductase.

3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase was investigated in different organic cosolvents by means of kinetic and calorimetric measurements, molecular dynamics simulations, and small-angle X-ray scattering. The combined experimental and theoretical techniques were essential to complement each other's limitations in the investigation of the complex interaction pattern between the enzyme, different solvent types, and concentrations. In this way, the underlying mechanisms for the loss of enzyme activity in different water-miscible solvents could be elucidated. These include direct inhibitory effects onto the active center and structural distortions.