Varying intensity of simulated infection partially affects the magnitude of the acute-phase immune response in the subterranean rodent Ctenomys talarum.

The acute phase response (APR), coordinated by a complex network of components of the immune and neuroendocrine systems, plays a key role in early immune defense. This response can be elicited by a wide variety of pathogens at different intensities (frequencies and doses), hence experimental immune challenges with antigen gradients makes it possible to evaluate sickness progression with a better representation of what occurs in natural systems. However, how infection intensity could shape the APR magnitude in wild species is still poorly understood. Here, the immune response was activated in the subterranean rodent Ctenomys talarum with a gradient of lipopolysaccharide (LPS) doses (0.5, 1, 1.5, and 2 mg/kg of body mass). Changes in body temperature, body mass, and energetic costs were evaluated over time. We also assessed cortisol levels, white blood cells counts and neutrophil: lymphocyte ratios, before and after injection. Results indicated that during the APR, C. talarum shows a hyperthermic response, which is maintained for 6 h, with slight differences among antigen doses in the pattern of thermal response and body mass change. A maximum increase in body temperature of 0.83°C to 1.63°C was observed during the first hour, associated with a metabolic cost that ranged from 1.25 to 1.41 ml O2 /gh. Although no clear effects of treatment were detected on leukocyte abundance, we found increments in neutrophil: lymphocyte ratios and gradual increases in cortisol levels corresponding to the intensity of simulated infection, which may indicate redistribution of immune cells and enhancement of immune function. An evident sickness syndrome was observed even at the lowest LPS dose that was characterized by an increase in body temperature, energy expenditure, and N: L ratio, as well as a dose-dependent increase in cortisol levels. Although in nature, other constraints and challenges could affect the magnitude and costs of immune responses, C. talarum mounts an effective APR with a low increase in their daily energy expenditure, regardless of LPS dose.

Personality underground: evidence of behavioral types in the solitary subterranean rodent Ctenomys talarum.

BACKGROUND: Animal personalities have been studied in a wide variety of taxa, but among rodents, available studies are relatively scarce and have focused mainly on social species. In this study, we evaluated the existence of personality in the solitary subterranean rodent Ctenomys talarum. Specifically, we aimed to test individual differences in behavior that are stable over time and context in males of C. talarum captured in the wild. METHODS: Our experimental design included two series of three behavioral tests each, carried out with a 35 day time interval. Each series included an Open Field test, a Social Encounter test, and an Open Field test with a predator stimulus. RESULTS: Of the total recorded behaviors, 55.55% showed temporal consistency. Principal component analysis of consistent behaviors grouped them into four dimensions that explain inter individual behavioral variability, in order of importance: activity, socioaversion, boldness and exploration. Therefore, our results suggest that the concept of animal personality is applicable to C. talarum and the dimensions found are in accordance with the ecological and behavioral characteristics of this species.

Differential antipredatory responses in the tuco-tuco (Ctenomys talarum) in relation to endogenous and exogenous changes in glucocorticoids.

Glucocorticoids participate in the behavioral and physiological responses generated under stressful circumstances coming from different sources-physical and/or psychological. In mammals, the increases of these hormones are mediated by the activation of the hypothalamic-pituitary-adrenal axis. This response occurs after exposure to novel and unpredictable situations that lead to the loss of homeostasis, for example, a direct encounter with predators or their cues. However, the relationship between the physiological and behavioral responses is still a complex issue in vertebrates. We evaluate the effects of an experimental manipulation of glucocorticoid levels on the generation of the behavioral and physiological response to stress by predation in the subterranean rodent C. talarum. We found that when tuco-tucos encountered predator cues-fur odor, and largely, immobilization-they responded physiologically by secreting cortisol. This response was accompanied by an associated behavioral response. However, when the increase in plasma cortisol originated exogenously by the injection of cortisol, a behavioral response was not observed. Finally, inhibition of glucocorticoids' synthesis was effective in weakening the behavioral effects produced by immobilization. In conclusion, in tuco-tucos, predator cues act as stress factors that trigger differential increases in plasma cortisol and a behavioral response associated with the appearance of anxiety states.

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.

Prenatal predatory stress in a wild species of subterranean rodent: Do ecological stressors always have a negative effect on the offspring?

In pregnant females, a failed predatory event not only induces individual responses but also represents a significant change in the developmental environment of the offspring, which may lead to modifications in their phenotype that may persist at different stages of life. We evaluate whether prenatal exposure to predatory cues affects anxiety behavior, behavioral response to predator cues, stress response to immobilization, and immune response to sheep red blood cells (SRBC) and phytohemagglutinin (PHA) in juveniles of the subterranean rodent Ctenomys talarum. We found that prenatal predator stress (PPS) (1) increased juvenile anxiety-like behaviors and the appearance of antipredator behaviors, (2) did not affect the response of offspring to predatory stressors, and (3) did not influence the physiological response of juveniles to stressors (immobilization) nor the immunological responses to SRBC and PHA challenges. This work shows the influence of PPS on the development of behavioral responses in the offspring, whom displayed a state of anxiety and behavioral changes associated with decreased locomotor activity and avoidance behaviors. Thus, these individuals prenatally exposed to predatory cues show behavioral adaptations that may contribute to avoid predators in the adult life.

Parasite infection negatively affects PHA-triggered inflammation in the subterranean rodent Ctenomys talarum.

Magnitude and effectiveness of immune responses vary greatly between and within species. Among factors reported to determine this variation, parasitism is a critical one, although controversial effects of parasites over immunological indices have been reported. Information regarding immune strategies in species with different life histories is crucial to better understand the role of immune defenses in an ecological and evolutionary context. Here, we examine the influence of the parasite community on immune responsiveness of a solitary subterranean rodent, Ctenomys talarum. To do this, we assessed the impact of the natural parasite community and the experimental infection with Eimeria sp. on the phytohemagglutinin (PHA)-response, as well as other immune, condition, nutrition, and stress parameters. PHA-triggered inflammation was similarly impaired by Eimeria sp. infection alone or co-occurring with a number of gastrointestinal nematodes. None of the other physiological parameters studied were affected by parasitism. This indicates that parasitism is a general key factor modulating immune responsiveness of the host, and in particular for C. talarum, it could explain the great inter-individual variation previously observed in the PHA-response. Thus, our results highlight the importance of taking the parasite community into account in ecoimmunological studies, particularly when using immunological indices.

Immune challenge but not dietary restriction affects spatial learning in the wild subterranean rodent Ctenomys talarum.

Several lines of evidence suggest that learning and triggering an immune response are both metabolically expensive and thus likely to be subject to nutritional trade-offs between them and other competing demands. Therefore, we evaluated if an immune challenge with a novel antigen affects spatial learning in the subterranean rodent Ctenomys talarum under two different dietary conditions. The results showed that immune-challenged animals were affected in their spatial learning capabilities, increasing the number of errors and marginally the time required to reach the goal of a complex labyrinth. No effect of the dietary restriction nor interaction between factors were observed. This work provides support for the existence of a trade-off between the costs of the immune defense and learning abilities, indicating that when investment is required to fight infection, fewer resources are available for learning. The absence of effect of nutritional condition on this trade-off suggests that other physiological processes, besides cognition, may be limited by the energetic resources necessary to the more immediately critical immune response.

PHA-induced inflammation is not energetically costly in the subterranean rodent Ctenomys talarum (tuco-tucos).

Immune activity has been proposed to be associated with substantial costs, due to trade-offs with other functions or activities that share common resources and contribute to an animal's fitness. However, direct estimates of the cost of mounting an immune response are few and have been performed mainly in birds. Thus, further work is needed to clarify the relative costs of different components of the immune system and the role of environmental and life-history traits in modulating the costs of resistance. Within the components of immunity, inflammation is considered to be associated with a larger energetic expenditure. Here, we evaluated the energetic cost of the inflammatory response to phytohemagglutinin (PHA) in a wild population of a subterranean rodent, Ctenomys talarum, and the trade-offs between immune activity and reproduction. C. talarum develops an inflammatory response to PHA, but contrary to our predictions, this response was not associated with an increase in oxygen consumption regardless of reproductive status or sex. Our study shows that an immune challenge may not always result in a detectable energetic cost. We discuss the possibility that other currencies could be underlying the cost, such as micro-or macronutrients requirements, autoimmunity or oxidative stress.

Assessing the energetic costs and trade-offs of a PHA-induced inflammation in the subterranean rodent Ctenomys talarum: immune response in growing tuco-tucos.

A traditional approach used to assess whether immune defense is costly is to explore the existence of trade-offs between immunity and other functions; however, quantitative studies of the energetic costs associated with the activation of the immune system are scarce. We assessed the magnitude of a PHA-triggered immune response and the associated energetic costs in 60-day old Ctenomys talarum. We expected that the magnitude of the macroscopic inflammatory response to PHA is lower in young tuco-tucos compared with that of adults, given the allocation of substantial energy to growth, and that the magnitude of the inflammation is lower in male pups compared to females, due to the higher investment in growth of the larger sex. Concomitantly, we expected that the pups challenged with PHA show an increase in oxygen consumption compared to control animals and that a positive association exists between magnitude of the PHA-induced inflammation and oxygen consumption. Contrary to what was expected, young tuco-tucos mounted a higher inflammatory response compared with adults and there were no differences in the magnitude of this response between sexes. The inflammatory response induced by a PHA injection did not represent a significant energetic cost for young tuco-tucos. There were no differences in oxygen consumption between PHA-injected and control animals, and tuco-tucos that mounted a higher inflammatory response to PHA did not show higher oxygen consumption. Energy expenditure, however, is not the only physiological cost involved in trade-offs between immune response and various functions of the organism, and other currencies are discussed.

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.

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.

Nutritional responses to different diet quality in the subterranean rodent Ctenomys talarum (tuco-tucos).

Nutritional response to different diet quality was examined in the subterranean rodent Ctenomys talarum (tuco-tuco). Animals maintained in captive conditions were fed with three plant species that differed in their fibre content. Tuco-tucos showed the ability to perform adjusts in short time lapse in response to diet quality; food ingestion, egestion and feces ingestion changed in animals under different plant species diets. Time budget, mainly time devoted to feeding and activity accompanied such changes. Coprophagy was practiced along the day and night following the arrhythmic activity pattern found for this species. Feces reingestion was not associated to resting. Furthermore, it was observed during fresh food ingestion, being pellets chewed. Soft and hard feces differed in morphological and nutritional characteristics.

Use of chemical communication by the subterranean rodent Ctenomys talarum (tuco-tuco) during the breeding season.

Solitary subterranean rodents with a low frequency of direct contact between conspecifics are expected to use chemical communication to coordinate social and reproductive behavior. We examined whether reproductive tuco-tucos (Ctenomys talarum) were able to discriminate the reproductive condition, sex, and source population of conspecifics by means of chemical cues contained in urine, feces, soiled shavings, or anogenital secretions. During preference tests in which animals had direct contact with these chemical cues, tuco-tucos were able to determine the reproductive condition of opposite sex conspecifics independent of the source of odor. When only olfactory cues were available, both sexes discriminated reproductive condition of opposite sex individuals using urine. Females were also able to discriminate the reproductive condition of males using soiled shavings. Females spent more time investigating male odors than female odors; except in the case of feces, breeding males spent similar amounts of time investigating male and female odors. No preferences were detected for opposite sex urine from members of an animal's own versus another population. The role of chemical cues in territory defense and breeding performance by this highly territorial subterranean rodent is discussed.

Bioenergetics of reproduction and pup development in a subterranean rodent (Ctenomys talarum).

The objective of this study was to evaluate the maternal costs of reproduction and pup development in the subterranean rodent Ctenomys talarum (Thomas 1898). Statistical differences were detected in whole-animal metabolic rates between nonreproductive and pregnant or lactating females. Whole-animal metabolic rates during pregnancy and lactation were 128% and 151% of the resting metabolic rate (RMR) observed in nonreproductive females. The total additional energy cost of reproduction (above the nonreproductive level) was similar for both the gestation and lactation periods. Mass-specific RMR revealed an upregulation of cell or tissue metabolism during lactation but not during gestation. The mass-specific metabolic rate of pups was 237% of the adults' metabolic rates. No differences were observed in body temperature among nonreproductive, pregnant, or lactating females. No differences were detected in body mass at birth among pups from litters with different numbers of nestlings. Pups increased their body temperature, reaching adult temperature at 30 d of age, when they were near weaning. Milk constituted the exclusive food for pups until they started eating solid food at 10 d old. Suckling time decreased with age of pups, and at the same time, mother chases directed toward their pups increased. These reproductive characteristics may contribute to successful existence in a subterranean habitat.