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.

Different regulation of cortisol and corticosterone in the subterranean rodent Ctenomys talarum: Responses to dexamethasone, angiotensin II, potassium, and diet.

When harmful environmental stimuli occur, glucocorticoids (GCs), cortisol and corticosterone are currently used to evaluate stress status in vertebrates, since their secretions are primarily associated to an increased activity of the hypothalamic-pituitaryadrenal (HPA) axis. To advance in our comprehension about GCs regulation, we evaluated the subterranean rodent Ctenomys talarum to assess cortisol and corticosterone response to (1) the negative feedback of the HPA axis using the dexamethasone (DEX) suppression test, (2) angiotensin II (Ang II), (3) potassium (K+) intake, and (4) different diets (vegetables, grasses, acute fasting). Concomitantly, several indicators of individual condition (body mass, neutrophil to lymphocyte ratio, blood glucose, triglycerides and hematocrit) were measured for diet treatments. Results confirm the effect of DEX on cortisol and corticosterone in recently captured animals in the field but not on corticosterone in captive animals. Data suggest that Ang II is capable of stimulating corticosterone, but not cortisol, secretion. Neither cortisol nor corticosterone were responsive to K+ intake. Cortisol levels increased in animals fed with grasses in comparison to those fed with vegetables while corticosterone levels were unaffected by diet type. Moreover, only cortisol responded to fasting. Overall, these results confirm that cortisol and corticosterone are not interchangeable hormones in C. talarum.

Diet effect on osmoregulation in the subterranean rodent Ctenomys talarum.

Water conservation requires osmoregulatory skills, sometimes limited by the environment and/or physiological and behavioral characteristics acquired along the evolutionary history of the species. Fossoriality had probably emerged as a survival mechanism to face increasing aridity, as suggested for Ctenomys, a genus that radiated to different environments. Ctenomys talarum (tuco-tuco) is an herbivorous subterranean rodent that lives in coastal grasslands inside humid burrows that reduce evaporation. However, their osmoregulatory mechanisms may be challenged by atmospheric variations when foraging aboveground and by the annual variability in dietary water and salt content. Then, it is of great interest to identify how much of this flexibility of C .talarum is attributed to physiological regulation. We analyzed the effect of water and salt content of diet on urinary, plasmatic, fecal and respiratory parameters. Tuco-tucos were not able to maintain their body weight under the offered monodiet, especially under the low hydrated diet, which explains its generalist and opportunistic foraging behavior. C. talarum mainly obtained water through food, whereas water metabolic production was negligible. Evaporative water loss did not vary between diets, but individuals under water restriction showed decreased fecal water loss and urine volume, high urine concentration but stable plasmatic osmolality and ionic concentration values. Under salt stress, urinary parameters remained relatively stable and high plasmatic osmolality was detected. Despite C. talarum produced more diluted urine than rodents from xeric environments, it is able to concentrate it 4 times above than the required at field even under the lowest water availability. This may be a characteristic associated with the evolutionary history of the species, which evolved in an arid context.

Thermogenic capacity in subterranean Ctenomys: Species-specific role of thermogenic mechanisms.

One way to understand ecological patterns of species is to determine their physiological diversity on a large geographic and/or temporal scales, in a context of hierarchical biodiversity framework. In particular, macrophysiological studies analyze how environmental factors affect the physiology and therefore the distribution of species. Subterranean species are an excellent model for evaluating the large-scale effects of ambient temperature (Ta) conditions on thermal physiology and distribution, due to their extensive use of burrows that provide a relatively thermal stable environment. Species belonging to the genus Ctenomys are all subterranean and endemic of South America. Cold induced maximum metabolic rate (MMR), basal metabolic rate (BMR) and non shivering thermogenesis (NST) were analyzed, as well as the expression of uncoupled proteins (UCP) in brown adipose tissue (BAT). Biogeographical variables appear to have no effect MMR experimentally induced by cold condition within Ctenomys. Also, mechanisms of heat production are species-specific, varying from a combination of ST and NST to a complete use of shivering mechanisms. This pattern is correlated at tissue level, since species that use only ST show a smaller interscapular BAT patch, not detectable presence of UCP1 and low COX activity. Thus, other factors, including body mass, that constrain cold induced MMR could affect thermogenic variability among Ctenomys. In the evolutionary timescale, if low O2 levels of burrows impose a ceiling in cold induced MMR, and ST is enhanced due to species-specific life history traits, such as digging effort, then the observed differences among Ctenomys species might be explained.

Expanding the actions of cortisol and corticosterone in wild vertebrates: A necessary step to overcome the emerging challenges.

We conducted a review of scientific articles published between 2000 and 2014 and evaluated how frequently various aspects of cortisol and corticosterone (CORT) actions have been considered in studies on wild vertebrates. Results show that (1) the notion that CORT are stress-responsive hormones is central in our theoretical frameworks and it is reflected by the fact that several articles refer to CORT as "stress hormones". (2) The large majority of studies do not contemplate the possibility of decrease and no change in CORT levels in response to chronic stressors. (3) Our ideas about CORT actions on energy balance are slanted towards the mobilization of energy, though there are several studies considering -and empirically addressing- CORT's orexigenic actions, particularly in birds. (4) The roles of CORT in mineral-water balance, though widely documented in the biomedical area, are virtually ignored in the literature about wild vertebrates, with the exception of studies in fish. (5) Adrenocorticotropic hormone (ACTH) independent regulation of CORT secretion is also very scarcely considered. (6) The preparative, permissive, suppressive and stimulatory actions of CORT, as described by Sapolsky et al. (2000), are not currently considered by the large majority of authors. We include an extension of the Preparative Hypothesis, proposing that the priming effects of baseline and stress-induced CORT levels increase the threshold of severity necessary for subsequent stimuli to become stressors. Studies on animal ecology and conservation require integration with novel aspects of CORT actions and perspectives developed in other research areas.

Effect of ambient temperature on evaporative water loss in the subterranean rodent Ctenomys talarum.

Subterranean rodents face unique thermoregulatory challenges. Evaporative water loss (EWL) is a crucial mechanism for maintaining heat balance in endotherms subjected to heat stress but also leads to potential dehydration. EWL depends on gradients of temperature and humidity between the surface of the individual and the surrounding environment. Underground burrows generally provide a stable water vapor saturated atmosphere which may impede evaporative heat loss (EHL). This will mainly occur when ambient temperature exceeds the upper limit of individual's thermoneutral zone, or when body temperature rises as result of digging activities. Here we evaluate the effect of ambient temperature on EWL and energy metabolism in the subterranean rodent Ctenomys talarum (tuco-tucos), which inhabits sealed burrows, but makes an extensive use of the aboveground environment. We observed that EWL is increased when ambient temperature rises above thermoneutrality; below this point, evaporation remains stable. Though EWL contributes to total heat loss by increasing ∼1.3 times at 35°C, dry thermal conductance is raised four times. In tuco-tucos' burrows both non-evaporative and, to some extent, evaporative and behavioral mechanisms are essential for body temperature regulation, preventing overheating at high ambient temperatures in a water vapor-saturated atmosphere.

Thermoregulatory development and behavior of Ctenomys talarum pups during brief repeated postnatal isolation.

In altricial mammals, the role of the mother and siblings throughout pup's early ontogeny is critical to determine "normal" development in neonates. It has been reported that variations in parental investment during pups' development affect thermoregulatory capacity, growth patterns, brain development and behavior during lifetime, such as spatial learning and memory in adults. Ctenomys talarum (tuco-tuco) is a solitary subterranean rodent, who inhabits complex burrows and exhibits developed spatial orientation abilities. Tuco-tuco's pups display an altricial development, spending more than 80% of the time in contact with the mother. Throughout weaning period, pups display active exploratory behavior and improvements in their spatial capabilities. Then, we determined the effect of repeated brief postnatal isolations on the acquisition of physiological thermoregulation and the development of spatial learning capabilities in tuco-tuco's pups. As it occurs in wild animals, daily brief isolations (30min) did not affect the acquisition of adult's body temperature nor resting metabolic rate's development pattern. Moreover, behavioral response and adult spatial abilities of isolated pups were similar to that observed in non-isolated ones. Then, during periods of mother's absence, minor physiological and behavioral adjustments, such as shivering and postural changes, are required to keep C. talarum pups within allostasis.

Cortisol and corticosterone exhibit different seasonal variation and responses to acute stress and captivity in tuco-tucos (Ctenomys talarum).

In this work we aimed to evaluate variations in plasma glucocorticoids (GCs, cortisol and corticosterone) levels throughout an annual cycle in free-living male tuco-tucos (Ctenomys talarum) and compare their responses to acute and chronic stressors (trapping, manipulation, immobilization, confinement in a novel environment, transference to captivity). In addition, we used leukocyte profiles to allow discrimination between basal and stress-induced seasonal changes in GC concentrations. Our results showed that cortisol and corticosterone are differently affected by environmental stimuli in C. talarum. Both hormones showed different patterns of variation in the field and responses to captivity. Moreover, only cortisol was responsive to acute stressors. Leukocyte profiles indicated that animals were unstressed in the field and therefore, that we were able to measure basal, stress-independent, fluctuations in GC levels. GC concentrations were low in comparison to values frequently reported for other mammals. Our results suggest differentiated physiological roles for cortisol and corticosterone in our study species and further emphasize the complexity of GC physiology in wild mammals.

Effect of diet quality and soil hardness on metabolic rate in the subterranean rodent Ctenomys talarum.

The present work is aimed to establish, in Ctenomys talarum, the physiological and behavioral adjustments undergone by individuals when they are allowed to dig burrows in soils with different hardness and fed with diets of different quality. For each soil-diet combination, we estimated: resting metabolic rate (RMR), body temperature (T(b)), body mass, digestibility, food consumption rate, transit time, reingestion rate, feces production and time devoted to feeding, resting, locomotor activity and coprophagy. Soil type and diet quality affected RMR, but response to soil hardness was verified later. Animals fed with high quality (HQ) diet showed similar body temperature irrespective of soil condition, while animals fed with low quality (LQ) diet showed lower T(b) under soft soil (SS). Individuals fed with LQ diet showed lower RMR and both, lower digestibility and high transit time of food than those fed with HQ diet. Moreover, increments in feeding and defecation rates were observed in the former group. Number of reingested feces did not differ between animals fed with diets of different quality. However, when incidence of reingestion was considered, animals fed with HQ diet showed higher values of feces ingestion. Either feeding, resting and activity patterns were arrhythmic. However, for animals fed with LQ diet a tendency to rhythmic coprophagy was observed and it could be considered as a way to optimize feeding. This study shows that RMR is limited by digestive efficiency which is influenced by diet quality, but also thermal stress may limit the conversion of assimilated energy into work and heat.

Decreased glucose tolerance but normal blood glucose levels in the field in the caviomorph rodent Ctenomys talarum: the role of stress and physical activity.

Hystricomorph rodents have a divergent insulin molecule with only 1-10% of the biological activity in comparison to other mammalian species. In this study, we used the subterranean rodent Ctenomys talarum as a model and performed blood glucose tolerance tests (GTTs) with trained and untrained individuals to evaluate blood glucose regulation and the possible role of physical activity as a compensatory mechanism. Additionally, we evaluated the variations in blood glucose during acute and chronic stress and gathered data in the field to evaluate natural-occurring variations in blood glucose levels. The GTTs showed that C. talarum have a diminished capacity of regulating blood glucose levels in comparison to other mammals and suggest that unexplored differences in the compensatory mechanisms, insulin structure and/or glucose transporters exist within species of hystricomorph rodents. However, blood glucose levels in the field stayed within the normal mammalian range. Physical activity did not prove to be a compensatory mechanism for blood glucose regulation. The individuals did not display important increases in blood glucose after acute stressors and managed to adequately regulate blood glucose during chronic stress. We suggest that the species may not face a selective pressure favoring a more tightly, mammalian like, capacity of regulating blood glucose levels.

Do subterranean mammals use the Earth's magnetic field as a heading indicator to dig straight tunnels?

Subterranean rodents are able to dig long straight tunnels. Keeping the course of such "runways" is important in the context of optimal foraging strategies and natal or mating dispersal. These tunnels are built in the course of a long time, and in social species, by several animals. Although the ability to keep the course of digging has already been described in the 1950s, its proximate mechanism could still not be satisfactorily explained. Here, we analyzed the directional orientation of 68 burrow systems in five subterranean rodent species (Fukomys anselli, F. mechowii, Heliophobius argenteocinereus, Spalax galili, and Ctenomys talarum) on the base of detailed maps of burrow systems charted within the framework of other studies and provided to us. The directional orientation of the vast majority of all evaluated burrow systems on the individual level (94%) showed a significant deviation from a random distribution. The second order statistics (averaging mean vectors of all the studied burrow systems of a respective species) revealed significant deviations from random distribution with a prevalence of north-south (H. argenteocinereus), NNW-SSE (C. talarum), and NE-SW (Fukomys mole-rats) oriented tunnels. Burrow systems of S. galili were randomly oriented. We suggest that the Earth's magnetic field acts as a common heading indicator, facilitating to keep the course of digging. This study provides a field test and further evidence for magnetoreception and its biological meaning in subterranean mammals. Furthermore, it lays the foundation for future field experiments.

Maximal thermogenic capacity and non-shivering thermogenesis in the South American subterranean rodent Ctenomys talarum.

Subterranean rodents inhabit closed tunnel systems that are hypoxic and hypercapnic and buffer aboveground ambient temperature. In contrast to other strictly subterranean rodents, Ctenomys talarum exhibits activity on the surface during foraging and dispersion and hence, is exposed also to the aboveground environment. In this context, this species is a valuable model to explore how the interplay between underground and aboveground use affects the relationship among basal metabolic rate (BMR), cold-induced maximum metabolic rate (MMR), shivering (ST), and non-shivering thermogenesis (NST). In this work, we provide the first evidence of the presence of NST, including the expression of uncoupling proteins in brown adipose tissue (BAT), and shivering thermogenesis in Ctenomys talarum, a species belonging to the most numerous subterranean genus, endemic to South America. Our results show no differences in BMR, cold-induced MMR, and NST between cold- (15 °C) and warm- (25 °C) acclimated individuals. Furthermore, thermal acclimation had no effect on the expression of mitochondrial uncoupling protein 1 (UCP1) in BAT. Only cytochrome c oxidase (COX) content and activity increased during cold acclimation. When interscapular BAT was removed, NST decreased more than 30%, whereas cold-induced MMR remained unchanged. All together, these data suggest that cold-induced MMR reaches a maximum in warm-acclimated individuals and so a probable ceiling in NST and UCP1 expression in BAT. Possible thermogenic mechanisms explaining the increase in the oxidative capacity, mediated by COX in BAT of cold-acclimated individuals and the role of ST in subterranean life habits are proposed.

Comparative energetics of the subterranean Ctenomys rodents: breaking patterns.

Subterranean mammals show lower mass-independent basal metabolic rates (BMRs). Several competing hypotheses were suggested to explain how microenvironmental conditions and underground life affect subterranean mammalian energetics. Two of these are the thermal stress and the cost-of-burrowing hypotheses. The thermal stress hypothesis posits that a lower mass-independent BMR reduces overheating in burrows where convective and evaporative heat loss is low, whereas the cost-of-burrowing hypothesis states that a lower mass-independent BMR may compensate for the high energy expenditure of digging. In this article, we assessed the relationship between BMR of Ctenomys and environmental variables through conventional statistics as well as independent contrasts. Moreover, we tested both the thermal stress and the cost-of-burrowing hypotheses at an interspecific level in a very homogeneous genus of subterranean rodents, the South American genus Ctenomys. We compared species from different geographic localities that have contrasting habitat conditions. We measured BMR through open-flow respirometry. After conventional as well as independent contrast analyses, our results support neither the thermal stress nor the cost-of-burrowing hypotheses. We observed that only body mass affects the variability in BMR. Contrasting climatic and soil conditions, habitat productivity, and net primary productivity were not correlated with BMR variability. We suggested that, because BMR and maximum metabolic rates (MMRs) are correlated, low BMRs among Ctenomys species could also be determined by factors that affect MMR rather than BMR.