Thermal biology in the Upper Galili Mountain blind mole rat (Nannospalax galili) and an overview of spalacine energetics.

Several hundred mammalian species thrive in complex burrow systems, which protect them from climatic extremes and predation. At the same time, it is also a stressful environment due to low food supply, high humidity, and, in some cases, a hypoxic and hypercapnic atmosphere. To face such conditions, subterranean rodents have convergently evolved low basal metabolic rate, high minimal thermal conductance and low body temperature. Although these parameters have been intensively studied in the last decades, such information is far from being well-known in one of the most studied groups of subterranean rodents, the blind mole rats of the genus Nannospalax. The lack of information is particularly noticeable for parameters such as the upper critical temperature and the width of the thermoneutral zone. In our study, we analysed the energetics of the Upper Galilee Mountain blind mole rat Nannospalax galili and found its basal metabolic rate of 0.84 ± 0.10 mL O2×g-1 × h-1, thermoneutral zone between 28 and 35 °C, mean Tb within the zone of 36.3 ± 0.6 °C, and minimal thermal conductance equal to 0.082 mL O2×g-1 × h-1 × C-1. Nannospalax galili is a truly homeothermic rodent well adapted to face lower ambient temperatures, because its Tb was stable down to the lowest temperature measured (10 °C). At the same time, a relatively high basal metabolic rate and relatively low minimal thermal conductance for a subterranean rodent of such body mass, and the difficulty of surviving ambient temperatures slightly above upper critical temperature, indicates problems with sufficient heat dissipation at higher temperatures. This can easily lead to overheating, that is relevant mainly during the hot-dry season. These findings suggest that N. galili can be threatened by ongoing global climate change.

Mammalian maxilloturbinal evolution does not reflect thermal biology.

The evolution of endothermy in vertebrates is a major research topic in recent decades that has been tackled by a myriad of research disciplines including paleontology, anatomy, physiology, evolutionary and developmental biology. The ability of most mammals to maintain a relatively constant and high body temperature is considered a key adaptation, enabling them to successfully colonize new habitats and harsh environments. It has been proposed that in mammals the anterior nasal cavity, which houses the maxilloturbinal, plays a pivotal role in body temperature maintenance, via a bony system supporting an epithelium involved in heat and moisture conservation. The presence and the relative size of the maxilloturbinal has been proposed to reflect the endothermic conditions and basal metabolic rate in extinct vertebrates. We show that there is no evidence to relate the origin of endothermy and the development of some turbinal bones by using a comprehensive dataset of µCT-derived maxilloturbinals spanning most mammalian orders. Indeed, we demonstrate that neither corrected basal metabolic rate nor body temperature significantly correlate with the relative surface area of the maxilloturbinal. Instead, we identify important variations in the relative surface area, morpho-anatomy, and complexity of the maxilloturbinal across the mammalian phylogeny and species ecology.

Body size, not species identity, drives body heating in alpine Erebia butterflies.

Efficient thermoregulation is crucial for animals living under fluctuating climatic and weather conditions. We studied the body heating of six butterfly species of the genus Erebia (Lepidoptera: Nymphalidae) that co-occur in the European Alps. We tested whether butterfly physical characteristics (body size, wing loading) are responsible for the inter-specific differences in body temperatures recorded previously under natural conditions. We used a thermal camera to measure body heating of wild butterfly individuals in a laboratory experiment with artificial light and heating sources. We revealed that physical characteristics had a small effect on explaining inter-specific differences in mean body temperatures recorded in the field. Our results show that larger butterflies, with higher weight and wing loading, heated up more slowly but reached the same asymptotic body temperature as smaller butterflies. Altogether, our results suggest that differences in body temperatures among Erebia species observed in the field might be caused mainly by species-specific microhabitat use and point towards an important role of active behavioural thermoregulation in adult butterflies. We speculate that microclimate heterogeneity in mountain habitats facilitates behavioural thermoregulation of adults. Similarly, microclimate structuring might also increase survival of less mobile butterfly life stages, i.e., eggs, larvae and pupae. Thus, landscape heterogeneity in management practices may facilitate long term survival of montane invertebrates under increased anthropogenic pressures.

Together or alone? Huddling energetic savings in three social mole-rat species of genus Fukomys. A dispersal perspective.

African mole-rats (Bathyergidae) are strictly subterranean rodents distributed in sub-Saharan Africa. Although the soil layer provides a temperature buffer, the temperature in their burrows is usually below their thermoneutral zone and thermogenesis is necessary to maintain a stable body temperature. In social bathyergids, an important mechanism for decreasing the thermoregulatory cost is social thermoregulation in the form of huddling. The effect of huddling may be of special importance during forming of a new family as only two adults are present and social species are known for higher heat losses from their bodies compared to solitary mole-rats. In our study, we measured the resting metabolic rate and energetic saving in three social bathyergid species which differ in body size. We compared animals that were housed individually and in pairs at two different ambient temperatures (Ta). At a temperature within their TNZ (Ta = 30 °C), no energetic savings were expected, whereas in Ta = 20 °C we expected energetic savings due to huddling. We found no energetic savings at 30 °C in any of the species, but almost 20% in the two small bodied Fukomys species F. micklemi and F. anselli at 20 °C. In the largest species, F. mechowii, no significant energetic savings were observed. Our results confirm the importance of huddling for the energetic balance of social mole-rats and show that huddling with one partner can bring substantial energetic savings, which can be allocated to other activities such as extension of established burrow systems or reproduction to increase the workforce and fulfill the purpose of dispersal.

MEDETOMIDINE-KETAMINE-BUPRENORPHINE ANESTHESIA OF THE SOLITARY SUBTERRANEAN CAPE DUNE MOLE-RAT (BATHYERGUS SUILLUS) AND THE CAPE MOLE-RAT (GEORYCHUS CAPENSIS).

A prospective, descriptive study was conducted to evaluate the safety and efficacy of a field-ready anesthetic drug combination of medetomidine-ketamine-buprenorphine for data logger implantation surgery or recheck in free-ranging Cape dune (Bathyergus suillus: n = 41) and Cape (Georychus capensis: n = 37) mole-rats. All anesthesia data were reported as mean (±standard deviation). Medetomidine-ketamine-buprenorphine doses were 0.1 (±0.03), 10.6 (±2.8), and 0.06 (±0.03) mg/kg, respectively, for Cape dune mole-rats; and 0.2 (±0.03), 19.4 (±4.0), and 0.14 (±0.03) mg/kg, respectively, for Cape mole-rats. Induction was calm and took 2.00 (range: 1.00-6.00) min for the Cape dune and 1.75 (range 1.25 to 8.16) min for Cape mole-rats. A surgical plane of anesthesia was achieved in most Cape dune mole-rats (92%) and Cape mole-rats (90%). The remainder required supplementation with a single intramuscular injection of ketamine (3-9 mg/kg) during surgery. Heart and respiratory rates were 149 (±37) beats and 24 (±8) breaths per minute, respectively, for Cape dune mole-rats and 179 (±40) beats and 25 (±10) and breaths per minute, respectively for Cape mole-rats. Surgical time for mole-rats ranged from 25 to 38 min. Recovery was calm and took 8.50 (range: 2.00-19.00) min for Cape dune mole-rats and 9.75 (range: 2.00-34.00) min for Cape mole-rats to recover. For recovery, atipamezole was administered intramuscularly at 0.5 (±0.15) mg/kg for Cape dune mole-rats and 1 (±0.15) mg/kg for Cape mole-rats. All mole-rats were returned to their original burrows within 48 h of recovery. The medetomidine-ketamine-buprenorphine combination induced a predictable, safe anesthesia in Cape dune and Cape mole-rats suitable for short intraabdominal surgery. This combination is suited to in situ studies where the use of a formal surgery or laboratory is not feasible.

Are southern African solitary mole-rats homeothermic or heterothermic under natural field conditions?

Abandoning of a stable body temperature (Tb), a phenomenon known as heterothermy, is an adaptation to cope mainly with a lack of food and water, especially in species inhabiting daily or seasonally variable environments. There is increasing evidence that African mammals avoid adverse conditions by heterothermy and eventually by entering torpor. Members of subterranean rodent family, the African mole-rats (Bathyergidae), are suitable candidates to study both phenomena, because of the diversity of their strategies in respect of maintaining stable Tb ranging from homeothermic species to a mammal with the most labile Tb, the naked mole-rat. Currently, there are field data on daily and seasonal Tb in one social species only and such information are lacking for any solitary mole-rat. In our study, we recorded yearly Tb in two solitary bathyergids, the Cape mole-rat Georychus capensis and the Cape dune mole-rat Bathyergus suillus from South Africa using intraperitoneally implanted dataloggers. Since this region is characterised by changing ecological characteristics, we expected either decreases of Tb within 24 h indicating daily torpor and/or longer-term decreases of Tb, which would indicate multiday torpor. Although we found seasonally phase shifted low amplitude daily Tb cycles, we did not find any remarkable and regular daily and/or seasonal Tb deviations, likely showing an absence of torpor in both species. Due to absence of this energy saving mechanism, we may speculate that both species could be vulnerable to ongoing global climatic change.

Heat dissipation in subterranean rodents: the role of body region and social organisation.

The relatively warm and very humid environment of burrows presents a challenge for thermoregulation of its mammalian inhabitants. It was found that African mole-rats dissipate body heat mainly through their venter, and social mole-rats dissipate more body heat compared to solitary species at lower temperatures. In addition, the pattern of the ventral surface temperature was suggested to be homogeneous in social mole-rats compared to a heterogeneous pattern in solitary mole-rats. To investigate this for subterranean rodents generally, we measured the surface temperatures of seven species with different degrees of sociality, phylogeny, and climate using infrared thermography. In all species, heat dissipation occurred mainly through the venter and the feet. Whereas the feet dissipated body heat at higher ambient temperatures and conserved it at lower ambient temperatures, the ventral surface temperature was relatively high in all temperatures indicating that heat dissipation to the environment through this body region is regulated mainly by behavioural means. Solitary species dissipated less heat through their dorsum than social species, and a tendency for this pattern was observed for the venter. The pattern of heterogeneity of surface temperature through the venter was not related to sociality of the various species. Our results demonstrate a general pattern of body heat exchange through the three studied body regions in subterranean rodents. Besides, isolated individuals of social species are less able to defend themselves against low ambient temperatures, which may handicap them if staying alone for a longer period, such as during and after dispersal events.

Evaporative water loss in seven species of fossorial rodents: Does effect of degree of fossoriality and sociality exist?

In terrestrial endotherms, evaporation is a significant mechanism of water loss in hot environments. Although water is passively lost by evaporation, individuals can regulate it at different levels. Inhabiting a relatively stable environment characterized by mild ambient temperature (Ta) and high humidity can ensure a balanced water budget. Many fossorial rodents are well adapted to live in such conditions. In this study, evaporative water loss (EWL) of fossorial rodent species with different degree of adaptations to underground life (from strictly subterranean to those with regular surface activity) was evaluated. By measuring EWL, the specific contribution of either evaporative or non-evaporative components of heat loss can be determined. With the exception of the silvery mole-rat (Heliophobius argenteocinereus), in all tested rodents EWL is relatively stable below and within the thermoneutral zone (TNZ). As Tas increase above TNZ, EWL increases as does total thermal conductance, but conductance increases several times more than EWL. In addition, non-evaporative routes seem to be more important than evaporative heat loss in the analyzed species. No clear pattern of EWL in relation to a species degree of fossoriality or sociality was detected. In this context, atmosphere of burrows could affect EWL, since the high humidity found inside tunnels can establish limits on evaporation to favor water rather than thermal balance.

Functional histology of the skin in the subterranean African giant mole-rat: thermal windows are determined solely by pelage characteristics.

Excavation of burrows is an extremely physically demanding activity producing a large amount of metabolic heat. Dissipation of its surplus is crucial to avoid the risk of overheating, but in subterranean mammals it is complicated due to the absence of notable body extremities and high humidity in their burrows. IR-thermography in a previous study on two species of African mole-rats revealed that body heat was dissipated mainly through the ventral body part, which is notably less furred. Here, we analyzed the dorsal and ventral skin morphology, to test if dermal characteristics could contribute to higher heat dissipation through the ventral body part. The thickness of the epidermis and dermis and the presence, extent and connectivity of fat tissue in the dermis were examined using routine histological methods, while vascular density was evaluated using fluorescent dye and confocal microscopy in the giant mole-rat Fukomys mechowii. As in other hitherto studied subterranean mammals, no subcutaneous adipose tissue was found. All examined skin characteristics were very similar for both dorsal and ventral regions: relative content of adipose tissue in the dermis (14.4 ± 3.7% dorsally and 11.0 ± 4.0% ventrally), connectivity of dermal fat (98.5 ± 2.8% and 95.5 ± 6.8%), vascular density (26.5 ± 3.3% and 22.7 ± 2.3%). Absence of large differences in measured characteristics between particular body regions indicates that the thermal windows are determined mainly by the pelage characteristics.

Higher resting metabolic rate in long-lived breeding Ansell's mole-rats (Fukomys anselli).

BACKGROUND: Reproduction is an energetically expensive process that supposedly impairs somatic integrity in the long term, because resources are limited and have to be allocated between reproduction and somatic maintenance, as predicted by the life history trade-off model. The consequence of reduced investment in somatic maintenance is a gradual deterioration of function, i.e. senescence. However, this classical trade-off model gets challenged by an increasing number of contradicting studies. Here we report about an animal model, which adds more complexity to the ongoing debate. Ansell's mole-rats are long-lived social subterranean rodents with only the founder pair reproducing, while most of their offspring remain in the parental burrow system and do not breed. Despite of a clear reproductive trade-off, breeders live up to twice as long as non-breeders, a unique feature amongst mammals. METHODS: We investigated mass-specific resting metabolic rates (msRMR) of breeders and non-breeders to gain information about the physiological basis underlying the reproduction-associated longevity in Ansell's mole-rats. We assessed the thermoneutral zone (TNZ) for breeders and non-breeders separately by means of indirect calorimetry. We applied generalized linear mixed-effects models for repeated measurements using the msRMR in the respective TNZs. RESULTS: TNZ differed between reproductive and non-reproductive Ansell's mole-rats. Contrary to classical aging models, the shorter-lived non-breeders had significantly lower msRMR within the thermoneutral zone compared to breeders. CONCLUSION: This is the first study reporting a positive correlation between msRMR and lifespan based on reproductive status. Our finding contradicts common aging theories, but supports recently introduced models which do not necessarily link reproductive trade-offs to lifespan reduction.

Asymmetric competition for nectar between a large nectar thief and a small pollinator: an energetic point of view.

There are two alternative hypotheses related to body size and competition for restricted food sources. The first one supposes that larger animals are superior competitors because of their increased feeding abilities, whereas the second one assumes superiority of smaller animals because of their lower food requirements. We examined the relationship between two unrelated species of different size, drinking technique, energy requirements and roles in plant pollination system, to reveal the features of their competitive interaction and mechanisms enabling their co-existence while utilising the same nectar source. We observed diurnal feeding behaviour of the main pollinator, the carpenter bee Xylocopa caffra and a nectar thief, the northern double-collared sunbird Cinnyris reichenowi on 19 clumps of Hypoestes aristata (Acanthaceae) in Bamenda Highlands, Cameroon. For comparative purpose, we established a simplistic model of daily energy expenditure and daily energy intake by both visitor species assuming that they spend all available daytime feeding on H. aristata. We revealed the energetic gain-expenditure balance of the studied visitor species in relation to diurnal changes in nectar quality and quantity. In general, smaller energy requirements and related ability to utilise smaller resources made the main pollinator X. caffra competitively superior to the larger nectar thief C. reichenowi. Nevertheless, sunbirds are endowed with several mechanisms to reduce asymmetry in exploitative competition, such as the use of nectar resources in times of the day when rivals are inactive, aggressive attacks on carpenter bees while defending the nectar plants, and higher speed of nectar consumption.

Extracellular adenosine mediates a systemic metabolic switch during immune response.

Immune defense is energetically costly, and thus an effective response requires metabolic adaptation of the organism to reallocate energy from storage, growth, and development towards the immune system. We employ the natural infection of Drosophila with a parasitoid wasp to study energy regulation during immune response. To combat the invasion, the host must produce specialized immune cells (lamellocytes) that destroy the parasitoid egg. We show that a significant portion of nutrients are allocated to differentiating lamellocytes when they would otherwise be used for development. This systemic metabolic switch is mediated by extracellular adenosine released from immune cells. The switch is crucial for an effective immune response. Preventing adenosine transport from immune cells or blocking adenosine receptor precludes the metabolic switch and the deceleration of development, dramatically reducing host resistance. Adenosine thus serves as a signal that the "selfish" immune cells send during infection to secure more energy at the expense of other tissues.

Surprisingly low risk of overheating during digging in two subterranean rodents.

Capacities for and constraints of heat dissipation are considered to be important factors governing maximum intensity and duration of physical activity. Subterranean mammals are endurance diggers, but because of lack of air currents in their burrows, high relative humidity and other physical constraints, the capacity of common mammalian cooling mechanisms underground is very limited. We analyzed surface and body core temperature changes after digging in soft and hard substrates in two species of African mole-rats (Bathyergidae, Rodentia); the social giant mole-rat Fukomys mechowii and the solitary silvery mole-rat Heliophobius argenteocinereus. As expected, we observed an increase of body core temperature in both species after digging in both substrates. Surprisingly, and contrary to our expectations, we observed remarkable decrease of mole-rats' surface temperature immediately after the end of the digging trials. This decrease was greater in soft and moister soil than that in hard and drier soil. Our results suggest that mole-rats may effectively avoid overheating in burrows by effective cooling while digging, especially in wet soil. This indicates that burrowing in soils moistened by rains could be easier than previously thought contributing thus to mole-rats success in challenging environment of subterranean burrows.

Social and Environmental Influences on Daily Activity Pattern in Free-Living Subterranean Rodents: The Case of a Eusocial Bathyergid.

Predictable daily activity patterns have been detected repeatedly even in mammals living in stable environments, as is the case for subterranean rodents. Whereas studies on activity of these rodents under laboratory conditions almost exclusively have concerned themselves with the influence of light, many field studies have revealed signs of an association between the activity pattern and daily fluctuations of temperature under the ground. This would assume that behavioral thermoregulation is probably involved. The only exceptions to the relationship between temperature and activity are 2 eusocial mole-rats of the genus Fukomys (Bathyergidae, Rodentia), which indicates that activity patterns could be affected also by social cues. To better understand how social and environmental factors influence the activity pattern in a eusocial mole-rat, we monitored the outside-nest activity in another species of this genus, the Ansell's mole-rat (Fukomys anselli), which has a relatively small body mass, high conductance, and more superficially situated burrows. Its daily activity had 1 prominent peak (around 1400 h), and it was tightly correlated with the temperature measured at depth of foraging burrows. Since F. anselli has high thermoregulatory requirements to maintain stable body temperature below the lower critical temperature, we conclude that the observed pattern is probably the result of minimizing the cost of thermoregulation. There were no significant differences in the daily activity patterns of breeding males and females and nonbreeders. Members of the same family group tended to have more similar activity patterns, but consistent activity synchronization between individuals was not proven. From the comparison of available data on all subterranean rodents, we assume that social cues in communally nesting mole-rats may disrupt (mask) temperature-related daily activity rhythms but probably only if the additional cost of thermoregulation is not too high, as it likely is in the Ansell's mole-rat.

Cost effective, robust, and reliable coupled separation techniques for the identification and quantification of phospholipids in complex biological matrices: application to insects.

The quantification of phospholipid classes and the determination of their molecular structures are crucial in physiological and medical studies. This paper's target analytes are cell membrane phospholipids, which play an important role in the seasonal acclimation processes of poikilothermic organisms. We introduce a set of simple and cost-effective analytical methods that enable efficient characterization and quantification of particular phospholipid classes and the identification and relative distribution of the individual phospholipid species. The analytical approach involves solid-phase extraction and high-performance thin-layer chromatography, which facilitate the separation of particular lipid classes. The obtained fractions are further transesterified to fatty acid methyl esters and subjected to gas chromatography coupled to flame ionization detection, which enables the determination of the position of double bonds. Phospholipid species separation is achieved by high-performance liquid chromatography with mass spectrometry, which gives information about the headgroup moiety and attached fatty acids. The total content of each phospholipids class is assessed by phosphorus determination by UV spectrophotometry. The simultaneous analysis of phosphorus, fatty acid residues, and phospholipid species provides detailed information about phospholipid composition. Evaluation of these coupled methods was achieved by application to an insect model, Pyrrhocoris apterus. High correlation was observed between fatty acid compositions as determined by gas chromatography and high-performance liquid chromatography analysis.

A seasonal difference of daily energy expenditure in a free-living subterranean rodent, the silvery mole-rat (Heliophobius argenteocinereus; Bathyergidae).

In seasonal climatic regimes, animals have to deal with changing environmental conditions. It is reasonable to expect that seasonal changes are reflected in animal overall energetics. The relation between daily energy expenditure (DEE) and seasonally variable ecological determinants has been studied in many free-living small mammals; however with inconsistent results. Subterranean mammals, i.e. fossorial (burrowing) mammals which live and forage underground, live in a seasonally and diurnally thermally stable environment and represent a suitable model to test seasonality in DEE in respect to seasonal changes, particularly those in soil characteristics and access to food supply. Both factors are affected by seasonal rainfall and are supposed to fundamentally determine activity of belowground dwellers. These ecological constraints are pronounced in some tropical regions, where two distinct periods, dry and rainy seasons, regularly alternate. To explore how a tropical mammal responds to an abrupt environmental change, we determined DEE, resting metabolic rate (RMR) and sustained metabolic scope (SusMS) in a solitary subterranean rodent, the silvery mole-rat, at the end of dry season and the onset of rainy season. Whereas RMR did not differ between both periods, mole-rats had 1.4 times higher DEE and SusMS after the first heavy rains. These findings suggest that rainfall is an important environmental factor responsible for higher energy expenditure in mole-rats, probably due to increased burrowing activity. SusMS in the silvery mole-rat is comparable to values in other bathyergids and all bathyergid values rank among the lowest SusMS found in endothermic vertebrates.

Cost of digging is determined by intrinsic factors rather than by substrate quality in two subterranean rodent species.

Searching for food by extensive digging is one of the most important aspects of life of subterranean rodents. We studied the effect of extrinsic (substrate quality) and intrinsic factors (sex and body mass) upon the cost of burrowing, expressed as digging metabolic rate (DMR) in two African mole-rat species (Bathyergidae, Rodentia) with distinct social structures. The sexually dimorphic giant mole-rat (Fukomys mechowii) is a highly social species, whereas the almost monomorphic silvery mole-rat (Heliophobius argenteocinereus) is a solitary bathyergid. Burrowing in F. mechowii was more costly (DMR was greater) than in H. argenteocinereus, but there was no difference in burrowing speed between both species. DMR within a particular species was dependent upon body mass, but independent of sex. Different substrate quality had no effect upon DMR in either species, yet it affected burrowing speed. We conclude that less effective digging in F. mechowii can be compensated by the joint workforce of other family members. Alternatively, H. argenteocinereus, being a more effective digger, can afford a solitary way of life.

Modulation of adrenergic receptors and adrenergic functions in cold adapted humans.

To specify the role of adrenoceptors in mediating adrenergic functions after adaptation of humans to cold, effect of administration of increasing concentrations of beta1 and beta2 adrenomimetics (Dobutamine, Bricanyl) on resting metabolic rate, heart rate, systolic blood pressure, rectal and skin temperatures of control humans and of cold adapted winter swimmers was studied. Increase in metabolic rate, mediated by beta1 and beta2 adrenomimetics, was attenuated after cold adaptation, indicating downregulation of beta1 and beta2 adrenoceptors. Since cold adapted humans have greater capacity of nonshivering thermogenesis, than that mediated by both beta1 and beta2 adrenoceptors, the role of other subtypes of adrenoceptors in mediating nonshivering thermogenesis is anticipated. Heart rate increased after administration of the beta2 agonist, but was not influenced by the beta1 agonist. The significance of beta2 adrenoceptors in mediating heart rate was depressed after cold adaptation. Data indicate that modifications of activity of beta adrenoceptors play crucial role in mechanisms responsible for adaptation of humans to cold.