Reproduction is a fundamental biological process, but it is also vulnerable to oxidative stress, which is a consequence of an imbalance between reactive oxygen species and antioxidant defences [...]

This study explores the metabolic factors that may aid in the division of labour in cooperatively breeding Damaraland mole-rat (Fukomys damarensis) colonies, particularly during digging activities. In these group-living mammals, both breeders and non-breeders participate in digging, among other cooperative tasks; however, non-breeding males and females (NBFs), often referred to as 'helpers', undertake this task at a higher frequency and engage in other energetically demanding activities more often than their breeding counterparts. We investigated how variation in glucose and lactate levels, two key energy substrates, relates to different levels of digging activity between breeders and non-breeders and how these metabolic patterns might underpin reproductive differences in activity and energy budgets. While both breeding females (BFs) and NBFs exhibited similar decreases in glucose levels after digging, lactate dynamics revealed a key distinction, NBFs experienced a significant drop in plasma lactate, suggesting lactate utilization. In contrast, BFs showed an increase in lactate, indicating its accumulation rather than utilization, potentially contributing to their reduced involvement in digging activity. These findings suggest that lactate recycling and metabolism may play a crucial role in sustaining prolonged physical exertion in NBFs, providing a potential physiological explanation for the division of labour in mole-rat colonies.

Enteropathogenic bacteria are a major cause of morbidity and mortality globally. Mouse models have been indispensable in advancing our understanding of infectious diseases caused by intestinal pathogens and in identifying physical, chemical and immunological barriers that limit these infections. However, there are significant differences between laboratory mice and human intestinal microbiota and immunobiology that underscore the need to develop other models that recapitulate the disease pathology and mucosal immune responses of human enteric diseases. Here we report how the pathogenic expansion of Citrobacter braakii in naked mole rats (NMRs) leads to colonic inflammation and epithelial injury that mimics pathological features of human hemorrhagic colitis. We observe mucosal erosions, ulcerations, depletion of goblet cells, extension of proliferative compartments to the surface of the glands, and active inflammation in the colonic lamina propria of infected NMRs. Without intervention, systemic inflammation associated with sepsis ensues in infected NMRs and results in high mortality. Interestingly, we demonstrate a strong therapeutic effect of probiotics comprising Lactobacillus, Bifidobacterium, Streptococcus salvarius subsp. thermophilus and Enterococcus faecium strains. Treatment with probiotics induces mucosal healing and restores intestinal homeostasis, including suppression of excessive proliferation of epithelial cells, replenishment of goblet cells and also has an anti-inflammatory effect. Taken together, we demonstrate that NMRs, beyond their use as an anti-ageing and disease-resistance model, can also be used to address disease mechanisms underlying infectious colitis, including disruptions in the mucosal barrier permeability, gut microbial ecology and in local and systemic immune regulation; and in testing functional probiotics strains as potential therapeutics. ### Competing Interest Statement The authors have declared no competing interest. South African Research Chair of Mammal Behavioural Ecology and Physiology, DST-NRF, GUN 64756 National Institute for Health Research (NIHR) Oxford Biomedical Research Centre South African Young Academy of Science Agency for Science, Technology and Research

In mammals, olfaction plays a key role in social behaviour, for example, in identifying mating opportunities and potential rivals. However, we still have a limited understanding of how social information is encoded in animal odours, including the social determinants of chemical similarity and diversity. Here, we used gas chromatography to analyse the chemical composition of swabs taken from the facial and anogenital regions of Damaraland mole-rats, Fukomys damarensis, a highly social subterranean mammal that relies almost exclusively on olfactory and tactile social cues. We found no sign of individual identity across the two body areas sampled; samples from the facial region and samples of the anogenital region from the same individual were not similar to each other, suggesting that these regions carry different information. However, chemical profiles varied significantly by sex and breeding status; female breeders differed from nonbreeders in their anogenital profiles and had higher chemical diversity in their facial profiles compared with both males and nonbreeders. Interestingly, we found no signals of social group identity. Instead, individual identity may be conveyed through signature mixes that are learned through frequent contact, rather than through specific odours associated with genetic kinship or social group membership. Our results highlight the complexity of chemical communication systems in social species and suggest that signals of group level identity are not necessary for behavioural responses based on group membership. (c) 2024 The Author(s). Published by Elsevier Ltd on behalf of The Association for the Study of Animal Behaviour. This is an open access article under the CC BY license (http://creativecommons.org/licenses/ by/4.0/).

Longevity is influenced by various factors, including fatty acid composition and free radical stress, which relate to the membrane pacemaker and rate of living hypotheses. While these aspects are well-documented in some long-lived species, they remain largely unexplored in tree squirrels. This study aimed to compare oxidative stress, antioxidant activity, nitrosative stress, and lipid composition between the long-lived Persian squirrel (Sciurus anomalus) and the short-lived Wistar rat across age cohorts (younger and older). Tissue homogenates from skin, liver, skeletal muscle, spleen, lung, and kidney were analysed for lipid composition (monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), arachidonic to linoleic acid ratio, peroxidation index, and unsaturation index. Oxidative, nitrosative, and antioxidant markers were assessed, including NADPH oxidase, superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase (GST), nitric oxide synthase, superoxide, hydrogen peroxide, nitric oxide, malondialdehyde, 4-hydroxynonenal, and total antioxidant capacity (TAC). Squirrels demonstrated higher GST activity, lower free radical stress, lower PUFA, and higher MUFA compared to rats. Antioxidant activities, except for TAC were negatively correlated with longevity. Older squirrels exhibited similar oxidative, nitrosative, and antioxidant profiles to younger squirrels, whereas younger rats displayed highly susceptible fatty acids, similar to older rats. The Persian squirrel's longevity appears closely linked to fatty acid composition and free radical resistance, likely due to increased GST activity. We propose GST's multifunctional role in reducing inflammation, enhancing immune response, providing disease resistance, and antioxidant activity contributes significantly to the longevity of the Persian squirrel.

Despite extensive studies on the physiology of subterranean rodents, there is comparatively little work documenting the energetics specifically associated with their locomotory energetics. The energetic cost associated with burrowing is great and, in part, explains why subterranean species often maintain their burrows and tunnels across generations. Indeed, the digging metabolic rate of five African mole-rats has been determined to be between three and five times higher than their respective resting metabolic rate, yet the energetic cost of non-digging locomotion (i.e. walking) has not been recorded. Digging in most subterranean species tends to lead to specialisation of the forelimbs and teeth, which may significantly affect the energetic cost associated with non-digging locomotion. Unlike many comparably sized burrowing and tunnelling mammals, African mole-rats appear, superficially at least, to have almost identical forelimbs and hindlimbs. This study explored the locomotory energetics associated with sustained submaximal locomotion (i.e. fast walking; 10 cm s-1) in two African mole-rat species (Fukomys damarensis and Heterocephalus glaber), utilising open-flow respirometry and a small animal treadmill. The mean locomotory energetic rate for F. damarensis was a near 1-fold increase (91.4%) above resting metabolic rate and a 2-fold increase (203.2%) for H. glaber. Net cost of transport was higher overall for H. glaber (2.9±0.6 ml O2 kg-1 m-1) than for F. damarensis (2.4±0.5 ml O2 kg-1 m-1). A trade-off likely exists between limb specialisation for digging and economic locomotion, and thus for most obligately subterranean species, locomotion represents an energetic investment.

Cooperation and group living have been suggested to facilitate survival in varying environments and under challenging conditions. However, group living may also be associated with costs, particularly in species where individuals within groups may compete for limited resources. The costs and benefits of cooperative group living on cooperatively breeding mammals in varying environments remain unclear. Here, we use data collected from wild, cooperatively breeding Natal mole-rats (Cryptomys hottentotus natalensis) inhabiting a seasonally varying environment to assess whether body condition changes between seasons and whether these changes are contingent upon group size. We demonstrate that the body condition of reproductive females improved with increasing helper number during both the benign summer and the harsher winter seasons. However, the body condition of other group members showed little dependency on group size or season. Only when including one extremely large group did the body condition of non-breeders weakly, albeit significantly, change depending on group size and seasonality. These results suggest that larger group sizes may yield some benefits for non-breeding group members during winter and may invoke some costs during summer. Group living in Natal mole-rats is likely promoted by a combination of collective foraging, thermoregulatory benefits, and potentially some indirect fitness benefits through improved body condition of the breeding female.

Animal movement paths display substantial complexity and variability, leading researchers to seek underlying rules that govern these patterns and mathematical models that best describe them. Using high-resolution (≥ 10 Hz) movement from 43 vertebrate species across diverse taxa, mass, and lifestyles, we show that movement paths are universally composed of straight-line steps interspersed with sharp turns, echoing a pattern documented for lower taxa such as bacteria. We report how these vertebrate ‘fundamental step lengths’ and ‘fundamental turn angles’, which are intrinsically different from the straight-line paths detailed in studies using low resolution position data, vary with species’ mass, lifestyle, behaviour, and environmental context. To explain these, we posit that animals inherently move in a straight line until sensory information signals a perceived better heading, which instigates a turn. The constellation of fundamental step lengths and turn angles over varying time intervals affects how well different models of animal movement (such as random walk or Lévy flight) fit lower resolution data. By examining turns as decision points, we can seek drivers of animal movement patterns and thereby work to predict future paths under varying conditions.

Selection pressures underpinning the evolution of mammalian sociality and body mass variation have spurred great interest for several decades. Because they inhabit a wide range of geographic localities and habitats, African mole-rat subspecies of Cryptomys hottentotus present a unique opportunity to further our understanding of intra- and interspecific variation of sociality and body mass in mammalian species. We compared the demographics and body masses among 4 C. hottentotus subspecies: the Mahali mole-rat, C. h. mahali; highveld mole-rat, C. h. pretoriae; Natal mole-rat, C. h. natalensis; and common mole-rat, C. h. hottentotus within the context of their respective microclimates and evolutionary history. We propose that all ancestral C. hottentotus subspecies were arid-adapted and thus formed large colonies (as found today in the Mahali mole-rat). However, as the subspecies dispersed to occupy habitats of varying aridity and temperature across South Africa, selection for a particular colony size range in each subspecies arose to provide an adaptive fitness benefit to survive in its habitat. Consequently, the Mahali mole-rat-which remained in a warm and arid environment-retained the largest mean colony size, followed by the Natal mole-rat, which-even though inhabiting a hyper-mesic environment-selected for increased colony sizes to offset the energy requirement of thermoregulation (through huddling) in the cooler montane environments they prefer. The common mole-rat and the highveld mole-rat possessed the smallest mean colony sizes, likely the result of inhabiting a mesic and warm environment. At the same time, body mass variation in the C. hottentotus complex is likely linked to colony size variation, with subspecies having the largest colonies possessing the lowest individual body mass. African mole-rats from the genus Cryptomys may hold the key to understanding proximate and ultimate factors behind mammalian social organization and body mass variation. Aridity and ambient temperature are considered strong selection pressures for colony-living and body mass variation in various subspecies of Cryptomys.

Whether the forelimb-digging apparatus of tooth-digging subterranean mammals has similar levels of specialization as compared to scratch-diggers is still unknown. We assessed the scapular morphology and forelimb musculature of all four solitary African mole rats (Bathyergidae): two scratch-diggers, Bathyergus suillus and Bathyergus janetta, and two chisel-tooth diggers, Heliophobius argenteocinereus and Georychus capensis. Remarkable differences were detected: Bathyergus have more robust neck, shoulder, and forearm muscles as compared to the other genera. Some muscles in Bathyergus were also fused and often showing wider attachment areas to bones, which correlate well with its more robust and larger scapula, and its wider and medially oriented olecranon. This suggests that shoulder, elbow, and wrist work in synergy in Bathyergus for generating greater out-forces and that the scapula and proximal ulna play fundamental roles as pivots to maximize and accommodate specialized muscles for better (i) glenohumeral and scapular stabilization, (ii) powerful shoulder flexion, (iii) extension of the elbow and (iv) flexion of the manus and digits. Moreover, although all bathyergids showed a similar set of muscles, Heliophobius lacked the m. tensor fasciae antebrachii (aiding with elbow extension and humeral retraction), and Heliophobius and Georychus lacked the m. articularis humeri (aiding with humeral adduction), indicating deeper morphogenetic differences among digging groups and suggesting a relatively less specialized scratch-digging ability. Nevertheless, Heliophobius and Bathyergus shared some similar adaptations allowing scratch-digging. Our results provide new information about the morphological divergence within this family associated with the specialization to distinct functions and digging behaviors, thus contributing to understand the mosaic of adaptations emerging in phylogenetically and ecologically closer subterranean taxa. This and previous anatomical studies on the Bathyergidae will provide researchers with a substantial basis on the form and function of the musculoskeletal system for future kinematic investigations of digging behavior, as well as to define potential indicators of scratch-digging ability. The muscles of scratch-diggers (Bathyergus spp.) show a higher mechanical advantage for forelimb flexion and shoulder stabilization than tooth-diggers. The glenohumeral and olecranon joints play fundamental roles as robust pivots to resist the action of larger muscles. Tooth-diggers lacked some muscles aiding with elbow extension and humeral retraction, suggesting deeper morphogenetic differences and comparatively diminished scratch-digging abilities.image

Oxidative stress plays a crucial role in mediating life-history processes, where it can compromise survival and reproduction through harmful alterations to DNA, lipids, and proteins. In this study, we investigated oxidative stress in Cape ground squirrels (Xerus inauris), a longer-lived African ground squirrel species with a high reproductive skew and unique life history strategies. We measured oxidative stress as total antioxidant capacity (TAC), total oxidant status (TOS), and an oxidative stress index (OSI) in blood plasma from individuals of approximately known ages. Our results reveal a distinct pattern of decreasing oxidative stress with age, consistent across both sexes. Females exhibited lower OSI and TOS levels than males. Males employing different life-history strategies, namely natal (staying at home), had significantly lower oxidative stress compared to the band (roaming male groups), likely due to variations in metabolic rate, activity, and feeding rates. However, both strategies exhibited reduced oxidative stress with age, though the underlying mechanisms require further investigation. We propose that selection pressures favoring survival contributed to the observed reduction in oxidative stress with age, potentially maximizing lifetime reproductive success in this species.

Arid-dwelling mammals are exposed to extreme daily temperatures, which puts these mammals at risk of overheating and dehydration, particularly while active, these risks are increased as the body size is reduced. Consequently, these small mammals often show strict nocturnality, confining their locomotor activity to the dark period of the day. One such animal is the Lesser Egyptian Jerboa (Jaculus jaculus), which occurs in the hyper-arid central regions of the Arabian Peninsula. In this study, twelve male J. jaculus were subjected to different light regimes. All the animals exhibited entrainment of their activity to the dark phase of each respective light regime. However, unlike other small arid dwelling mammals, J. jaculus confined most of its activity during the last few hours of the night rather than the initial hours of the night, a pattern which is consistent even under constant DD conditions. The last few hours of the night correlate to the coolest experienced temperatures in summer in the wild and, therefore, the ideal time for a very small mammal to be active. Larger small mammals, which may be less at risk of overheating, can afford to be active during the warmer periods of the night, namely the initial hours.

In the current study we employed immunohistochemical techniques to investigate the localisation of three calcium-binding proteins (parvalbumin, calbindin, and calretinin) in 10 neuronal structures of the cerebellar cortex (stellate cells, basket cells, parallel fibres, climbing fibres, Purkinje cells, granule cells, Golgi type II cells, Lugaro neurons, unipolar brush neurons, and mossy fibres) in 143 species from across the mammalian radiation. Most often, parvalbumin was localised in the neural structures of the molecular and Purkinje cell layers but was absent in the granule cell layer. Calbindin was most often immunolocalised in the neural structures of the Purkinje cell layer and mossy fibres, whereas calretinin was most often immunolocalised in the climbing fibres of the molecular layer and all neural structures of the granule cell layer. Despite this general consistency, variations in the localisation of these three calcium-binding proteins were found in every lineage, and almost every species, the one exception being the western tree hyrax that showed the full suite of most often observed calcium-binding protein chemoarchitecture for the mammalian cerebellar cortex. These consistencies and variances in the calcium-binding protein chemoarchitecture of the cerebellar cortex of mammals may play significant roles in the species-specific learning and refining of motor, perceptual, and cognitive skills and capacities required to survive in the environments they inhabit.

Subterranean mammals representing a single subspecies occurring along an aridity gradient provide an appropriate model for investigating adaptive variation in thermal physiology with varying levels of precipitation and air temperature. This study examined the thermal physiological adaptations of common mole-rats (Cryptomys hottentotus hottentotus) across five populations along an aridity gradient, challenging the expectation that increased aridity would lead to reduced metabolic rate, lower body temperatures and broader thermoneutral zones. No significant, consistent differences in metabolic rate, body temperature or thermal conductance were observed between populations, suggesting uniform thermoregulatory mechanisms across habitats. Instead, behavioural strategies such as huddling and torpor may play a more prominent role than physiological adaptations in managing temperature regulation and water balance. The study also observed osmoregulatory differences, with populations employing distinct behavioural cooling strategies in response to water availability. These results underscore the need for further research into the responses of subterranean species to climate change, particularly in understanding how increasing global temperatures and aridification might influence species distribution if they lack the physiological capacity to adapt to future climatic conditions.

Light is considered the primary entrainer for mammalian biological rhythms, including locomotor activity (LA). However, mammals experience different environmental and light conditions, which include those predominantly devoid of light stimuli, such as those experienced in subterranean environments. In this study, we investigated what environmental cue (light or ambient temperature (Ta)) is the strongest modulator of circadian rhythms, by using LA as a proxy, in mammals that experience a lifestyle devoid of light stimuli. To address this question, this study exposed a subterranean African mole-rat species, the Damaraland mole-rat (Fukomys damarensis), to six light and Ta cycles in different combinations. Contrary to previous literature, when provided with a reliable light cue, Damaraland mole rats exhibited nocturnal, diurnal, or arrhythmic LA patterns under constant Ta. While under constant darkness and a 24-hour Ta cycle mimicking the burrow environment, all mole-rats were most active during the coolest 12-hour period. This finding suggests that in a subterranean environment, which receives no reliable photic cue, the limited heat dissipation and energy constraints during digging activity experienced by Damaraland mole-rats make Ta a reliable and consistent "time-keeping" variable. More so, when providing a reliable light cue (12 light: 12 dark) to Damaraland mole-rats under a 24-hour Ta cycle, this study presents the first evidence that cycles of Ta affect the LA rhythm of a subterranean mammal more strongly than cycles of light and darkness. Once again, Damaraland mole-rats were more active during the coolest 12-hour period regardless of whether this fell during the light or dark phase. However, conclusive differentiation of entrainment to Ta from that of masking was not achieved in this study, and as such, we have recommended future research avenues to do so.

ABSTRACTAlthough the mammalian cerebral cortex is most often described as a hexalaminar structure, there are cortical areas (primary motor cortex) and species (elephants, cetaceans, and hippopotami), where a cytoarchitecturally indistinct, or absent, layer 4 is noted. Thalamocortical projections from the core, or first order, thalamic system terminate primarily in layers 4/inner 3. We explored the termination sites of core thalamocortical projections in cortical areas and in species where there is no cytoarchitecturally distinct layer 4 using the immunolocalization of vesicular glutamate transporter 2, a known marker of core thalamocortical axon terminals, in 31 mammal species spanning the eutherian radiation. Several variations from the canonical cortical column outline of layer 4 and core thalamocortical inputs were noted. In shrews/microchiropterans, layer 4 was present, but many core thalamocortical projections terminated in layer 1 in addition to layers 4 and inner 3. In primate primary visual cortex, the sublaminated layer 4 was associated with a specialized core thalamocortical projection pattern. In primate primary motor cortex, no cytoarchitecturally distinct layer 4 was evident and the core thalamocortical projections terminated throughout layer 3. In the African elephant, cetaceans, and river hippopotamus, no cytoarchitecturally distinct layer 4 was observed and core thalamocortical projections terminated primarily in inner layer 3 and less densely in outer layer 3. These findings are contextualized in terms of cortical processing, perception, and the evolutionary trajectory leading to an indistinct or absent cortical layer 4.

Abstract The cardiac conduction system in large carnivores, such as the African lion (Panthera leo), represents a significant knowledge gap in both veterinary science and in cardiac electrophysiology. Short QT intervals have been reported from zoo‐kept, anaesthetized lions, and our goal was to record the first ECGs from wild, conscious lions roaming freely, and compare them to zoo‐kept lions under the hypothesis that short QT is unique to zoo‐kept lions. Macroscopic and histological examinations were performed on heart tissue removed from nine healthy zoo lions. ECGs were recorded from the nine anaesthetized zoo‐kept lions, and from 15 anaesthetized and conscious wild lions in Africa. Our histological and topographical description of the lion's heart matched what has previously been published. In conscious lions, the ECG recordings revealed a mean heart rate of 70 ± 4 beats/min, with faster heart rates during the night. PQ and QT intervals were heart rate dependent in the conscious lions. Interestingly, QT intervals recorded in wild lions were markedly longer than QT intervals from zoo lions (398 ± 40 vs. 297 ± 9 ms, respectively; P < 0.0001). Anaesthesia or heart rate did not account for this difference. We provide a comprehensive description of the cardiac anatomy and electrophysiology of wild and zoo‐kept lions. QT intervals were significantly shorter in zoo lions, suggesting functional disparities in cardiac electrophysiology between wild and zoo‐kept lions, potentially related to physical fitness. These findings underscore the plasticity of cardiac electrophysiology and may be of value when reintroducing endangered species into the wild and when managing lions in human care.

Our research represents the first extensive study of the breeding behaviour and related environmental triggers of reproduction in the African giant pouch rat (Cricetomys gambianus Waterhouse, 1840) in and around the equator throughout a whole year. We measured the gross morphology and detailed histology of both female and male rats, along with plasma steroid hormone levels. Contrary to other tropical-dwelling small mammals, the African giant pouch rat is a seasonal breeder; however, rainfall is not to be the primary cue of reproduction. Our study suggests that ambient temperature and photoperiod are the primary environmental cues of reproduction, with breeding occurring during the cooler months of the year, namely in the dry season. During the wet and hot season, which succeeds the dry season, there is an increase in the availability of quality food, which results in nursing mothers and weaned pups achieving a positive energy balance and increased body condition. This, in turn, increases pregnancy success and offspring survival. Climate change, particularly global warming, could harm the reproduction of African giant pouched rats as rising temperatures in and around the equator, including Tanzania, may impact their circannual reproductive cycle.