Differential locomotor activity responses to day-time light intensity in juvenile and adult solitary Cape mole-rats, Georychus capensis (Rodentia: Bathyergidae).

The Cape mole-rat (Georychus capensis) is a solitary, strictly subterranean rodent that is responsive to light and entrains to photic cues despite having a reduced visual system. Circadian entrainment is maintained throughout life, but age can alter the amplitude of the response and re-entrainment time. Mole-rats are long-lived for their size which raises questions regarding the robustness of their circadian rhythms and how impacts their locomotor activity rhythms. The locomotor activity rhythms of juvenile and adult Cape mole-rats were investigated. They were exposed to pre-experimental and post-experimental control cycles under fluorescent lights, six 12 h light:12 h dark cycles of decreasing intensities and a constant dark cycle (DD). All animals exhibited more activity during the dark phases of all light regimes. Juveniles were more active than adults and displayed more variable activity during both the light and dark phases. Adults exhibited relatively stable levels of activity under all experimental conditions, whereas juvenile activity decreased as the light intensity was reduced. The amplitude of Cape mole-rat rhythms was consistently low, but similar across light regimes and between adults and juveniles. Cape mole-rats have functional circadian systems, are primarily nocturnal and respond differentially to light intensity depending on their age. Light intensity does not affect the locomotor activity responses of Cape mole-rats in a predictable manner, and could indicate more complex interactions with light wavelengths. The circadian systems of juveniles appear to be more sensitive than those of adults, although the mechanism of the light response remains unclear.

Dim light at night affects the locomotor activity of nocturnal African pygmy mice (Mus minutoides) in an intensity-dependent manner.

Rodents are integral components of ecosystems as they provide several important ecosystem services. Despite their importance as prey, pollinators and seed distributors, African rodents are largely understudied. The effect of anthropogenic changes such as artificial light at night extends past urban areas to peri-urban and rural habitats, and can have profound effects on entire ecosystems. We investigated the effect of dim light at night (dLAN) on the locomotor activity rhythms of the African pygmy mouse (Mus minutoides). Pygmy mice showed a dramatic, intensity-dependent reduction in their locomotor activity when subjected to dLAN, which was accompanied by a delay in the activity onset. We also considered masking responses with a dark pulse (DP) during the day and a light pulse at night. All animals became inactive in response to a light pulse during the night, whereas approximately half of the animals showed activity during a DP in the day. Our results suggest that the African pygmy mouse is highly sensitive to light and that their activity is strongly masked by light. In their natural environment, vegetation could shield pygmy mice against high light levels; however, other anthropogenic disturbances can alter the behaviour of these animals and could affect their survival.

Flexibility in body temperature rhythms of free-living natal mole-rats (Cryptomys hottentotus natalensis).

The subterranean niche is a specialised and particularly challenging environment to obtain direct physiological and behavioural measurements from free-living animals. Rhythmicity has been examined in many mole-rat species in the laboratory, but field reports are relatively scarce. We implanted Natal mole-rats with temperature loggers in summer and winter to record core body temperature (Tb), before releasing the animals again. Animals were recaptured after two months to recover the loggers. Natal mole-rats can maintain rhythmicity of their Tb in their natural habitat and display seasonal differences in their Tb rhythms. During winter mole-rats have unimodal Tb rhythms, whereas in summer many animals have bimodal Tb patterns, which may be related to temperatures in the foraging burrows close to the soil surface. Individuals from the same colonies frequently exhibited similar rhythms, especially in the larger colonies, which may indicate social entrainment of rhythms. Males and females differ in their seasonal variation of Tb, with males having more variation in winter, while the opposite was true for females. Entire colonies may undergo sporadic torpor bouts, presumably to conserve energy, but the trigger for these events is unknown. This is the first report of torpor occurring in an African mole-rat species. The ability to respond to environmental and social cues, while rhythms remain flexible, can provide an adaptive advantage to animals living in challenging and energetically demanding habitats.

Oxidative stress in response to heat stress in wild caught Namaqua rock mice, Micaelamys namaquensis.

Modelling of anthropogenic induced climate suggests more frequent and severe heatwaves in the future, which are likely to result in the mass die-off of several species of organisms. Oxidative stress induced by severe heat stress has previously been associated with a reduction in animal cognitive performance, depressed reproduction and lower life expectancy. Little is known about the non-lethal consequences of species should they survive extreme heat exposure. We investigated the oxidative stress experienced by the Namaqua rock mouse, a nocturnal rodent, using two experimental heat stress protocols, a 6 hour acute heat stress protocol without access to water and a 3-day heatwave simulation with ad libitum water. Oxidative stress was determined in the liver, kidney and brain using malondialdehyde (MDA) and protein carbonyl (PC) as markers of oxidative damage, and superoxide dismutase (SOD) and total antioxidant capacity (TAC) as markers of antioxidant defence. Incubator heat stress (heat and dehydration stress) was brought about by increasing the body temperatures of animals to 39-40.8 °C for 6 hours. Following incubator heat stress, significantly higher levels of MDA were observed in the liver. Dehydration did not explain the variation in oxidative markers and is likely a combined effect of thermal and dehydration stress. Individual body mass was significantly negatively correlated to kidney SOD and lipid peroxidation. A heatwave was simulated using a temperature cycle that would naturally occur during a heatwave in the species' local habitat, with a maximal ambient temperature of 38 °C. Following the simulated heatwave, SOD activity of the kidney demonstrated significantly lowered activity suggesting oxidative stress. Current heat waves in this species have the potential of causing oxidative stress. Heat and dehydration stress following exacerbated temperatures are likely to incur significant oxidative stress in multiple tissues demonstrating the importance of water availability to allow for rehydration to prevent oxidative stress.

Laterality in the Cape mole-rat, Georychus capensis.

Behavioural lateralization, the differential use one side of the body, and/or the bilateral use of sensory organs or limbs, is common in many vertebrates. One way in which behavioural lateralization can be detected in animals is through turning biases, which is an inherent preference to either turn left or right. Mole-rats are a unique group of mammals that demonstrate a wide range of social organizations ranging from solitary to eusociality. Behavioural asymmetry has not previously been investigated in mole-rats. In this study, captive and wild solitary Cape-mole rats (Georychus capensis) were investigated for individual (relative laterality (LR)) and population-level (absolute laterality (LA)) laterality. Mole-rats in the captive group were in the laboratory for at least one year, whereas the wild group were captured and experimented on within 2 weeks of capture. Animals were placed in a Y-maze facing away from the centre of the maze, and the turn towards the centre of the maze was evaluated to determine individual turning biases. Lateralized individual turning biases were more apparent in wild (7/9), compared to captive (3/10) individuals. Both captive and wild populations demonstrated a left bias, which was higher in wild animals, but not significantly so. Cape mole-rats are extremely xenophobic and aggressive, and this aggressive behaviour may underlie the turning biases in these animals, as aggression is primarily a right hemisphere dominant process. The reduced lateralization observed in captive animals may be due to a reduced need for these behaviours as a result of different environments in captivity.

Heat and dehydration induced oxidative damage and antioxidant defenses following incubator heat stress and a simulated heat wave in wild caught four-striped field mice Rhabdomys dilectus.

Heat waves are known for their disastrous mass die-off effects due to dehydration and cell damage, but little is known about the non-lethal consequences of surviving severe heat exposure. Severe heat exposure can cause oxidative stress which can have negative consequences on animal cognition, reproduction and life expectancy. We investigated the current oxidative stress experienced by a mesic mouse species, the four striped field mouse, Rhabdomys dilectus through a heat wave simulation with ad lib water and a more severe temperature exposure with minimal water. Wild four striped field mice were caught between 2017 and 2019. We predicted that wild four striped field mice in the heat wave simulation would show less susceptibility to oxidative stress as compared to a more severe heat stress which is likely to occur in the future. Oxidative stress was determined in the liver, kidney and brain using malondialdehyde (MDA) and protein carbonyl (PC) as markers for oxidative damage, and superoxide dismutase (SOD) and total antioxidant capacity (TAC) as markers of antioxidant defense. Incubator heat stress was brought about by increasing the body temperatures of animals to 39-40.8°C for 6 hours. A heat wave (one hot day, followed by a 3-day heatwave) was simulated by using temperature cycle that wild four striped field mice would experience in their local habitat (determined through weather station data using temperature and humidity), with maximal ambient temperature of 39°C. The liver and kidney demonstrated no changes in the simulated heat wave, but the liver had significantly higher SOD activity and the kidney had significantly higher lipid peroxidation in the incubator experiment. Dehydration significantly contributed to the increase of these markers, as is evident from the decrease in body mass after the experiment. The brain only showed significantly higher lipid peroxidation following the simulated heat wave with no significant changes following the incubator experiment. The significant increase in lipid peroxidation was not correlated to body mass after the experiment. The magnitude and duration of heat stress, in conjunction with dehydration, played a critical role in the oxidative stress experienced by each tissue, with the results demonstrating the importance of measuring multiple tissues to determine the physiological state of an animal. Current heat waves in this species have the potential of causing oxidative stress in the brain with future heat waves to possibly stress the kidney and liver depending on the hydration state of animals.

Temporal flexibility in activity rhythms of a diurnal rodent, the ice rat (Otomys sloggetti).

Diurnality in rodents is relatively rare and occurs primarily in areas with low nighttime temperatures such as at high altitudes and desert areas. However, many factors can influence temporal activity rhythms of animals, both in the field and the laboratory. The temporal activity patterns of the diurnal ice rat were investigated in the laboratory with, and without, access to running wheels, and in constant conditions with running wheels. Ice rats appeared to be fundamentally diurnal but used their running wheels during the night. In constant conditions, general activity remained predominantly diurnal while wheel running was either nocturnal or diurnal. In some animals, entrainment of the wheel running rhythm was evident, as demonstrated by free-running periods that were different from 24 h. In other animals, the wheel running activity abruptly switched from nocturnal to subjective day as soon as the animals entered DD, and reverted back to nocturnal once returned to LD, suggesting the rhythms were masked by light. Wheel running rhythms appears to be less robust and more affected by light compared to general activity rhythms. In view of present and future environmental changes, the existence of more unstable activity rhythms that can readily switch between temporal niches might be crucial for the survival of the species.

Locomotor activity in field captured crepuscular four-striped field mice, Rhabdomys dilectus and nocturnal Namaqua rock mice, Micaelamys namaquensis during a simulated heat wave.

Activity of animals is influenced by ambient temperature and increasing temperatures brought about by climate change may impose a heat stress risk. Previous studies investigating the effect of heat waves on activity usually measure animals at different, but constant temperatures, however, rarely are they studied under a natural temperature cycle. General activity, behavioural flexibility and frequency of water drinking counts during a normal day, hot day and a simulated heat wave temperature cycle were studied in the crepuscular four-striped field mouse, Rhabdomys dilectus, and the nocturnal Namaqua rock mouse, Micaelamys namaquensis. Both R. dilectus and M. namaquensis showed typical daily locomotor activity under control conditions. During the heat wave, peak activity times changed for R. dilectus, but both species exhibited higher bouts of activity for the heat wave during the day compared to the control, which was accompanied by an increased amount of time spent drinking water. The increased activity during the heat wave is likely due to enhanced water requirements and potentially a form of behavioural thermoregulation as animals may be uncomfortable and try to move to cooler areas. Thus, in the absence of a typical microclimate, access to water may allow rodents to overcome heat stress from extreme temperatures without having to shift their temporal active times.

Trading new neurons for status: Adult hippocampal neurogenesis in eusocial Damaraland mole-rats.

Diversity in social structures, from solitary to eusocial, is a prominent feature of subterranean African mole-rat species. Damaraland mole-rats are eusocial, they live in colonies that are characterized by a reproductive division of labor and a subdivision into castes based on physiology and behavior. Damaraland mole-rats are exceptionally long lived and reproductive animals show delayed aging compared to non-reproductive animals. In the present study, we described the hippocampal architecture and the rate of hippocampal neurogenesis of wild-derived, adult Damaraland mole-rats in relation to sex, relative age and social status or caste. Overall, Damaraland mole-rats were found to have a small hippocampus and low rates of neurogenesis. We found no correlation between neurogenesis and sex or relative age. Social status or caste was the most prominent modulator of neurogenesis. An inverse relationship between neurogenesis and social status was apparent, with queens displaying the lowest neurogenesis while the worker mole-rats had the most. As there is no natural progression from one caste to another, social status within a colony was relatively stable and is reflected in the level of neurogenesis. Our results correspond to those found in the naked mole-rat, and may reflect an evolutionary and environmentally conserved trait within social mole-rat species.

Orexinergic neuron numbers in three species of African mole rats with rhythmic and arrhythmic chronotypes.

In the present study, orexinergic cell bodies within the brains of rhythmic and arrhythmic circadian chronotypes from three species of African mole rat (Highveld mole rat-Cryptomys hottentotus pretoriae, Ansell's mole rat--Fukomys anselli and the Damaraland mole rat--Fukomys damarensis) were identified using immunohistochemistry for orexin-A. Immunopositive orexinergic (Orx+) cell bodies were stereologically assessed and absolute numbers of orexinergic cell bodies were determined for the distinct circadian chronotypes of each species of mole rat examined. The aim of the study was to investigate whether the absolute numbers of identified orexinergic neurons differs between distinct circadian chronotypes with the hypothesis of elevated hypothalamic orexinergic neurons in the arrhythmic chronotypes compared with the rhythmic chronotypes. We found statistically significant differences between the circadian chronotypes ofF. anselli, where the arrhythmic group had higher mean numbers of hypothalamic orexin neurons compared with the rhythmic group. These differences were observed when the raw data was compared and when the raw data was corrected for body mass (M(b)) and brain mass (M(br)). For the two other species investigated, no significant differences were noted between the chronotypes, although a statistically significant difference was noted between all rhythmic and arrhythmic individuals of the current study when the counts of orexin neurons were corrected for M(b)--the arrhythmic individuals had larger numbers of orexin cells.

Reproductive suppression and the seasonality of reproduction in the social Natal mole-rat (Cryptomys hottentotus natalensis).

Natal mole-rats are social subterranean rodents which exhibit a reproductive division of labour. Reproduction is confined to a single breeding female and one or more males; the remainder of the colony members is reproductively suppressed by the presence of the breeding animals. Apart from the discovery that female Natal mole-rats are induced ovulators, little is known about the reproductive biology of this species. Natal mole-rats are closely related to common and highveld mole-rats, both of which are induced ovulators and seasonal breeders. We therefore postulated that reproduction in Natal mole-rats is seasonally regulated. However, the results indicate that dominant Natal mole-rats are able to reproduce in the winter as well as in the summer. Furthermore, the increment in plasma LH in response to GnRH does not show marked seasonal differences in reproductive or non-reproductive mole-rats of either sex. Nevertheless, in all reproductive categories the level of plasma LH is significantly higher in the winter than in the summer. Seasonality in plasma LH levels dissociated from seasonality in breeding seems paradoxical. Further investigations will be required to elucidate this finding. We also investigated the processes underlying socially regulated reproduction in this species by determining basal and GnRH-evoked plasma LH in reproductive and non-reproductive animals of each sex in both seasons. The results failed to identify neuroendocrine differences consistent with an inhibited reproductive state in subordinates of either sex. Thus, the present findings suggest that behavioural interactions and/or inbreeding avoidance are the principal factors underlying suppression of reproduction in subordinate Natal mole-rats.

An immunohistochemical study of the gonadotrophin-releasing hormone 1 system in solitary Cape mole-rats, Georychus capensis, and social Natal mole-rats, Cryptomys hottentotus natalensis.

Mole-rat species within the family Bathyergidae exhibit a wide range of reproductive strategies and social systems. Various forms of reproductive suppression are displayed within this family: in the solitary species, breeding is suspended for part of the year and in the social species, reproduction is suppressed in subordinate animals. This study investigated the gonadotrophin-releasing hormone 1 (GnHR-1) systems of breeding and non-breeding solitary Cape mole-rats and social Natal mole-rats for possible inter- and/or intra-species differences. In both species, GnRH-1 cell bodies are predominantly in the medial septum region of the diagonal band or the preoptic area, with relatively few in the mediobasal hypothalamus; a dense concentration of GnRH-1-immunoreactive (ir) processes is present in the region of the organum vasculosum of the lamina terminalis. In Cape mole-rats, GnRH-1-ir processes are particularly dense within the lateral margins of the median eminence, which is enfolded by a large pars tuberalis of the pituitary gland. Natal mole-rats display GnRH-1-ir processes across the breadth of the median eminence, which is abutted by a relatively small pars tuberalis. There are more GnRH-1-ir cell bodies in Natal mole-rats than in Cape mole-rats ( approximately 720 vs. approximately 420). No significant differences were found in the number, distribution or size of GnRH-1-ir cell bodies according to season in Cape mole-rats or according to reproductive status or sex in Natal mole-rats. In female and male Natal mole-rats, GnRH-1-immunoreactivity in the median eminence is less dense in the reproductive animals; no such difference was found in Cape mole-rats between the breeding and non-breeding seasons. These immunohistochemical results are discussed in the light of earlier studies which identified no functional neuroendocrine impediments underlying regulated reproduction in either Cape or Natal mole-rats. The cumulative findings suggest that the principal factors determining seasonal or socially induced suppression of reproduction in these species are behavioral rather than neuroendocrine.

Energetics reveals physiologically distinct castes in a eusocial mammal.

Eusociality, which occurs among mammals only in two species of African mole-rat, is characterized by division of labour between morphologically distinct 'castes'. In Damaraland mole-rats (Cryptomys damarensis), colony labour is divided between 'infrequent worker' and 'frequent worker' castes. Frequent workers are active year-round and together perform more than 95% of the total work of the colony, whereas infrequent workers typically perform less than 5% of the total work. Anecdotal evidence suggests that infrequent workers may act as dispersers, with dispersal being limited to comparatively rare periods when the soil is softened by moisture. Here we show that infrequent workers and queens increase their daily energy expenditure after rainfall whereas frequent workers do not. Infrequent workers are also fatter than frequent workers. We suggest that infrequent workers constitute a physiologically distinct dispersing caste, the members of which, instead of contributing to the work of the colony and helping the queen to reproduce, build up their own body reserves in preparation for dispersal and reproduction when environmental conditions are suitable.

Seasonal energetics of the Hottentot golden mole at 1500 m altitude.

Winter is an energetically stressful period for small mammals as increasing demands for thermoregulation are often coupled with shortages of food supply. In sub-tropical savannah, Hottentot golden moles (Ambysomus hottentottus longiceps) forage throughout the year and for long periods of each day. This may enable them to acquire sufficient resources from an insectivorous prey base that is both widely dispersed and energetically costly to obtain. However, they also inhabit much cooler regions; how their energy budgets are managed in these areas is unknown. We measured the daily energy expenditure (DEE), resting metabolic rate (RMR) and water turnover (WTO) of free-living golden moles during both winter and summer at high altitude (1500 m). We used measurements of deuterium dilution to estimate body fat during these two periods. DEE, WTO and body mass did not differ significantly between seasons. However, RMR values were higher during the winter than the summer and, in the latter case were also lower than allometric predictions. Body fat was also higher during the winter. Calculations show that during the winter they may restrict activity to shorter, more intense periods. This, together with an increase in thermal insulation, might enable them to survive the cold.

Fos expression in the suprachiasmatic nucleus in response to light stimulation in a solitary and social species of African mole-rat (family Bathyergidae).

Mole-rats are strictly subterranean rodents that are rarely exposed to environmental light. They are well adapted to their environment and have reduced eyes and a severely regressed visual system. It has been shown, however, that mole-rats do exhibit endogenous circadian rhythms that can be entrained, suggesting an intact and functional circadian system. To determine whether light is the entraining agent in these animals, Fos expression in response to light pulses at different circadian times was investigated to obtain phase response curves. Light is integrated effectively in the suprachiasmatic nucleus of the Cape mole-rat (Georychus capensis), and Fos expression is gated according to the phase of the circadian clock. The Fos response in the Cape mole-rat was comparable to that of aboveground rodents. In contrast, the highveld mole-rat (Cryptomys hottentotus pretoriae) was less sensitive to light and did not show a selective Fos response according to the phase of the circadian cycle. Social species appear to be less sensitive to light than their solitary counterparts, which compares well with results from locomotor activity studies.

The chronobiology of the Natal mole-rat, Cryptomys hottentotus natalensis.

The Natal mole-rat, Cryptomys hottentotus natalensis, rarely, if ever, is exposed to external light cues because it occurs in completely sealed tunnel systems. As a result, their classical visual system is regressed, and therefore, their circadian system is expected proportionally to be expanded. Locomotor activity was investigated under a number of different photic regimes. Nine of the 12 mole-rats exhibited endogenous circadian rhythms of locomotor activity under constant darkness, with a mean free run period of 24.13 h (range 23.93-24.13 h), with these animals entrained to a light-dark cycle (12 L:12 D). Because C. hottentotus natalensis are able to entrain their locomotor activity to an external light source, light must reach the suprachiasmatic nucleus (SCN), suggesting a functional circadian clock. A clear day-night rhythm of melatonin secretion in animals housed under a neutral photoperiod (12 L:12 D) was observed, with higher melatonin concentrations in the dark compared with the light phase. The rhythm was maintained after the animals were transferred to either continuous light (LL) or dark (DD), suggesting that the endogenous rhythm was maintained under acute exposure to light and dark. However, under DD, the rhythm appeared to shift slightly, potentially as a result of the rhythm free running. These results show that C. hottentotus natalensis has endogenous rhythms of both locomotor activity and melatonin secretion, which are modulated by light.