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.

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.

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.

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.

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.

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.

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.

A new cue for torpor induction: charcoal, ash and smoke.

Recent work has shown that the use of torpor for energy conservation increases after forest fires in heterothermic mammals, probably in response to the reduction of food. However, the specific environmental cues for this increased torpor expression remain unknown. It is possible that smoke and the novel substrate of charcoal and ash act as signals for an impending period of starvation requiring torpor. We therefore tested the hypothesis that the combined cues of smoke, a charcoal/ash substrate and food shortage will enhance torpor expression in a small forest-dwelling marsupial, the yellow-footed antechinus (Antechinus flavipes), because like other animals that live in fire-prone habitats they must effectively respond to fires to ensure survival. Activity and body temperature patterns of individuals in outdoor aviaries were measured under natural environmental conditions. All individuals were strictly nocturnal, but diurnal activity was observed shortly after smoke exposure. Overall, torpor in females was longer and deeper than that in males. Interestingly, while both males and females increased daily torpor duration during food restriction by >2-fold as anticipated, a combination of food restriction and smoke exposure on a charcoal/ash substrate further increased daily torpor duration by ∼2-fold in both sexes. These data show that this combination of cues for torpor induction is stronger than food shortage on its own. Our study provides significant new information on how a small forest-dwelling mammal responds to fire cues during and immediately after a fire and identifies a new, not previously recognised, regulatory mechanism for thermal biology in mammals.

Post-fire recovery of torpor and activity patterns of a small mammal.

To cope with the post-fire challenges of decreased availability of food and shelter, brown antechinus (Antechinus stuartii), a small marsupial mammal, increase the use of energy-conserving torpor and reduce activity. However, it is not known how long it takes for animals to resume pre-fire torpor and activity patterns during the recovery of burnt habitat. Therefore, we tested the hypothesis that antechinus will adjust torpor use and activity after a fire depending on vegetation recovery. We simultaneously quantified torpor and activity patterns for female antechinus from three adjacent areas: (i) the area of a management burn 1 year post-fire, (ii) an area that was burned 2 years prior, and (iii) a control area. In comparison to shortly after the management burn, antechinus in all three groups displayed less frequent and less pronounced torpor while being more active. We provide the first evidence that only 1 year post-fire antechinus resume pre-fire torpor and activity patterns, probably in response to the return of herbaceous ground cover and foraging opportunities.

Copper-Induced Topology Switching and Thrombin Inhibition with Telomeric DNA G-Quadruplexes.

The topological diversity of DNA G-quadruplexes may play a crucial role in its biological function. Reversible control over a specific folding topology was achieved by the synthesis of a chiral, glycol-based pyridine ligand and its fourfold incorporation into human telomeric DNA by solid-phase synthesis. Square-planar coordination to a CuII ion led to the formation of a highly stabilizing intramolecular metal-base tetrad, substituting one G-tetrad in the parent unimolecular G-quadruplex. For the Tetrahymena telomeric repeat, CuII -triggered switching from a hybrid-dominated conformer mixture to an antiparallel topology was observed. CuII -dependent control over a protein-G-quadruplex interaction was shown for the thrombin-tba pair (tba=thrombin-binding aptamer).

Cool echidnas survive the fire.

Fires have occurred throughout history, including those associated with the meteoroid impact at the Cretaceous-Palaeogene (K-Pg) boundary that eliminated many vertebrate species. To evaluate the recent hypothesis that the survival of the K-Pg fires by ancestral mammals was dependent on their ability to use energy-conserving torpor, we studied body temperature fluctuations and activity of an egg-laying mammal, the echidna (Tachyglossus aculeatus), often considered to be a 'living fossil', before, during and after a prescribed burn. All but one study animal survived the fire in the prescribed burn area and echidnas remained inactive during the day(s) following the fire and substantially reduced body temperature during bouts of torpor. For weeks after the fire, all individuals remained in their original territories and compensated for changes in their habitat with a decrease in mean body temperature and activity. Our data suggest that heterothermy enables mammals to outlast the conditions during and after a fire by reducing energy expenditure, permitting periods of extended inactivity. Therefore, torpor facilitates survival in a fire-scorched landscape and consequently may have been of functional significance for mammalian survival at the K-Pg boundary.

Friends with benefits: the role of huddling in mixed groups of torpid and normothermic animals.

Huddling and torpor are widely used for minimizing heat loss by mammals. Despite the questionable energetic benefits from social heterothermy of mixed groups of warm normothermic and cold torpid individuals, the heterothermic Australian sugar glider (Petaurus breviceps) rests in such groups during the cold season. To unravel why they might do so, we examined torpor expression of two sugar glider groups of four individuals each in outside enclosures during winter. We observed 79 torpor bouts during 50 days of observation and found that torpor bouts were longer and deeper when all individuals of a group entered torpor together, and therefore infer that they would have saved more energy in comparison to short and shallow solitary torpor bouts. However, all gliders of either group only expressed torpor uniformly in response to food restriction, whereas on most occasions at least one individual per group remained normothermic. Nevertheless, the presence of warm gliders in mixed groups also appears to be of energetic advantage for torpid individuals, because nest box temperature was negatively correlated with the number of torpid gliders, and normothermic individuals kept the nest temperature at a value closer to the threshold for thermoregulatory heat production during torpor. Our study suggests that mixed groups of torpid and normothermic individuals are observed when environmental conditions are adverse but food is available, leading to intermediate energy savings from torpor. However, under especially challenging conditions and when animals are starving, energy savings are maximized by uniform and pronounced expression of torpor.

Can hibernators sense and evade fires? Olfactory acuity and locomotor performance during deep torpor.

Increased habitat fragmentation, global warming and other human activities have caused a rise in the frequency of wildfires worldwide. To reduce the risks of uncontrollable fires, prescribed burns are generally conducted during the colder months of the year, a time when in many mammals torpor is expressed regularly. Torpor is crucial for energy conservation, but the low body temperatures (T b) are associated with a decreased responsiveness and torpid animals might therefore face an increased mortality risk during fires. We tested whether hibernators in deep torpor (a) can respond to the smell of smoke and (b) can climb to avoid fires at T bs below normothermic levels. Our data show that torpid eastern pygmy-possums (Cercartetus nanus) are able to detect smoke and also can climb. All males aroused from torpor when the smoke stimulus was presented at an ambient temperature (T a) of 15 °C (T b ∼18 °C), whereas females only raised their heads. The responses were less pronounced at T a 10 °C. The first coordinated movement of possums along a branch was observed at a mean T b of 15.6 °C, and animals were even able to climb their prehensile tail when they reached a mean T b of 24.4 °C. Our study shows that hibernators can sense smoke and move at low T b. However, our data also illustrate that at T b ≤13 °C, C. nanus show decreased responsiveness and locomotor performance and highlight that prescribed burns during winter should be avoided on very cold days to allow torpid animals enough time to respond.

The importance of mammalian torpor for survival in a post-fire landscape.

Wildfires have increased in frequency and intensity worldwide with climate change as a main driving factor. While a number of studies have focused on population changes in regard to fires, there are essentially no quantitative data on behavioural and physiological adjustments that are vital for the persistence of individuals during and after fires. Here we show that brown antechinus, a small insectivorous marsupial mammal, (i) endured a prescribed fire in situ, (ii) remained in their scorched home range despite unburned areas nearby, and (iii) substantially increased post-fire torpor use and thus reduced foraging requirements and exposure to predators. Hence, torpor is a physiological adaptation that, although not quantified in this context previously, appears to play a key role in post-fire survival for this and other heterothermic species.

The hustle and bustle of city life: monitoring the effects of urbanisation in the African lesser bushbaby.

Urbanisation has become a severe threat to pristine natural areas, causing habitat loss and affecting indigenous animals. Species occurring within an urban fragmented landscape must cope with changes in vegetation type as well as high degrees of anthropogenic disturbance, both of which are possible key mechanisms contributing to behavioural changes and perceived stressors. We attempted to elucidate the effects of urbanisation on the African lesser bushbaby, Galago moholi, by (1) recording activity budgets and body condition (body mass index, BMI) of individuals of urban and rural populations and (2) further determining adrenocortical activity in both populations as a measure of stress via faecal glucocorticoid metabolite (fGCM) levels, following successful validation of an appropriate enzyme immunoassay test system (adrenocorticotropic hormone (ACTH) challenge test). We found that both sexes of the urban population had significantly higher BMIs than their rural counterparts, while urban females had significantly higher fGCM concentrations than rural females. While individuals in the urban population fed mainly on provisioned anthropogenic food sources and spent comparatively more time resting and engaging in aggressive interactions, rural individuals fed almost exclusively on tree exudates and spent more time moving between food sources. Although interactions with humans are likely to be lower in nocturnal than in diurnal species, our findings show that the impact of urbanisation on nocturnal species is still considerable, affecting a range of ecological and physiological aspects.

Nonshivering thermogenesis in the African lesser bushbaby, Galago moholi.

The capacity for nonshivering thermogenesis (NST) plays an important role during arousal from torpid states. Recent data on heterotherms inhabiting warmer regions, however, suggest that passive rewarming reduces the need of metabolic heat production during arousal significantly, leading to the question: to what extent do subtropical or tropical heterotherms depend on NST? The African lesser bushbaby, Galago moholi, enters torpid states as an emergency response only, but otherwise stays normothermic throughout the cold and dry winter season. In addition, this species shows unusual rewarming difficulties during arousal from torpor on cold days. We therefore examined the seasonal adjustments of the capacity for NST of naturally acclimatized G. moholi by stimulation with noradrenaline (NA) injection. Dissection of two adult female bushbabies revealed that G. moholi possesses brown adipose tissue, and NA treatment (0.5 mg kg(-1), s.c.) induced a significant elevation in oxygen consumption compared with control (saline) injection. However, the increase in oxygen consumption following injection of NA was not significantly different between winter and summer. Our results show that the ability to produce heat via NST seems to be available throughout the year and that G. moholi is able to change NST capacity within a very short time frame in response to cold spells. Together with results from studies on other (Afro-)tropical heterotherms, which also indicate low or even absent seasonal difference in NST capacity, this raises the question of whether the definition of NST needs to be refined for (Afro-)tropical mammals.

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.

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.