Stress in an underground empire.

The naked mole-rat (Heterocephalus glaber) and the Damaraland mole-rat (Fukomys damarensis) live in large colonies in underground tunnel systems in sub-Saharan Africa. Most members of the colonies are suppressed from reproduction and they are unlikely to reproduce during their lifetime. Only one female and a small number of males reproduce. This extreme cooperative social system has fascinated researchers since the naked mole-rat was first described as eusocial. Despite much research into the mechanisms of social suppression, the exact mechanisms are still unclear. Much evidence points towards high glucocorticoid concentrations caused by agonistic behaviour by the breeding female suppressing reproduction of non-breeders, but laboratory studies have not found any differences in glucocorticoids between breeders and non-breeders. There is, however, considerable evidence from field studies and other social mole-rats that social stress may indeed be an important factor of social suppression in social mole-rats and that those mechanisms are affected by the stability of the colony and environmental conditions. This review aims to provide a summary of the current knowledge of the relationship between environmental conditions, colony stability, glucocorticoids and reproductive suppression in social mole-rat species and suggests some avenues for future research.

Repeatability of daily profiles of baseline glucocorticoid metabolites measured in the urine and faeces of eastern rock sengis (Elephantulus myurus).

Repeatability of hormone concentrations is of great interest for studies investigating the evolution of hormonal traits. Particularly the repeatability of glucocorticoids (GCs) in response to a stressor is frequently investigated, but often only point (initial and/or response value), or single measures are used. A new method takes into account the entire individual hormone profile and generates an individual profile repeatability (PR) score. The method was developed for response profiles, but it may also be valuable for baseline values in species with diurnal changes in hormone concentrations. GCs are determined in a variety of matrices, and repeatability can vary considerably depending on the matrix. We investigated the repeatability of baseline GC metabolite (GCM) concentrations measured in urine (uGCM) and faeces (fGCM) of captive eastern rock sengis (Elephantulus myurus) using the more traditional linear mixed model approach and the PR method. GCMs were assessed over 24 h and measurements were repeated twice with two weeks between replicates. A diurnal rhythm in GCM concentrations associated with the activity period of the sengis was found in urine, but not in the faeces. Urinary GCM concentrations exhibited a moderate repeatability, whereas the repeatability of fGCM concentrations was low. Urinary GCM concentrations and their repeatability differed between the sexes; with higher concentrations and lower PR scores in females. No such sex differences were apparent for fGCM concentrations and the PR score was not able to characterise repeatability of fGCM concentrations, which were lacking a distinct profile. The PR score enabled a successful quantification of the repeatability of the diurnal uGCM profiles. Hormone profile, sex and sample matrix can affect hormonal traits considerably and the results may be obscured if these factors are not carefully considered.

Effect of colony disruption and social isolation on naked mole-rat endocrine correlates.

The social environment of animals can have profound implications on their behaviour and physiology. Naked mole-rats (Heterocephalus glaber) are highly social with complex dominance hierarchies that influence both stress- and reproduction-related hormones. Homeostasis may be affected by aggressive interactions, colony instability and social isolation. Furthermore, naked mole-rat colonies are characterised by a marked reproductive skew; a single female and few males are reproductively active while other colony members are reproductively suppressed. Thus, there are distinct differences in related hormone concentrations between reproductively active and non-active animals; however, this changes when non-reproductive individuals are removed from the colony. We investigated the effects of social isolation and colony disruption on plasma cortisol and progesterone concentrations in non-breeding naked mole-rats. During colony disruption, we found a significant increase in cortisol concentrations in females removed from the colony for social isolation (experimental) as well as in females that remained in the colony (control). Cortisol concentrations were reduced in both groups after experimental animals were paired up. No changes in cortisol concentrations were observed in control or experimental males after removal from the colony or pairing. This suggests that the females, but not the males, found colony disruption and social isolation stressful. Upon removal from the colony, both control and experimental females showed a small increase in progesterone, which returned to basal levels again in the control animals. Experimental females showed a dramatic spike in progesterone when they were paired with males, indicating reproductive activation. The sex difference in the stress responses may be due to the stronger reproductive suppression imposed on females, or the increased likelihood of dispersal for males. It is clear that the social environment reflects on the endocrine correlates of animals living in a colony, and that the colony structure may affect the sensitivity of the animals to changes in their environment.

Sex and dose-dependent responses of urinary and faecal glucocorticoid metabolite concentrations following an ACTH challenge in eastern rock sengis (Elephantulus myurus).

Non-invasive methods for measuring glucocorticoids and their metabolites are frequently used in ecological, behavioural and physiological studies of mammals. Using faeces, urine and other matrices for such a measurement has considerable advantages in comparison to more traditional methods, but also requires thorough validation of the methods used. Eastern rock sengis (Elephantulus myurus) are fascinating African mammals and the non-invasive monitoring of the adrenocortical activity opens up new opportunities to study their biology. We were able to validate two assays for measuring urinary (uGCM) and faecal glucocorticoid metabolite (fGCM) concentrations in this species using a dose-dependent challenge with adrenocorticotropic hormone (ACTH). A higher concentration of ACTH elicited higher uGCM and fGCM concentrations in both males and females. Interestingly, uGCM and fGCM concentrations and the responses to ACTH were higher in females than in males and small changes in faecal glucocorticoid metabolites could not be reliably detected in males. In contrast to ACTH, a saline injection did not result in an increase in uGCM or fGCM concentrations. The study also provided insight into when responses to a stressor are likely to be detected in the urine and faeces of sengis and opens up new opportunities to study the stress physiology of this and other sengi species. It further emphasises the importance of thoroughly validating non-invasive methods for measuring hormones in both sexes of a species and for incorporating dose-dependent approaches.

Changes in prolactin, cortisol and testosterone concentrations during queen succession in a colony of naked mole-rats (Heterocephalus glaber): a case study.

Colonies of naked mole-rats (Heterocephalus glaber, NMRs) are characterised by an extreme skew in lifetime reproductive success with only one female and one to three male consorts in a colony. The rest of the individuals in a colony are reproductively suppressed and much research has been focussed on elucidating that mechanism. The dopamine system and prolactin have recently been implicated in the suppression of reproduction of subordinate NMRs. To investigate the changes in prolactin during the removal of an aged reproductive female (queen) and succession of a new queen, blood samples were collected during different stages of queen removal: before queen removal, after separation, but in olfactory contact with the queen and after the total removal of the queen. Further, plasma cortisol and testosterone concentrations were determined. The colony appeared unstable prior to queen removal as indicated by high concentrations of cortisol and testosterone and lack of successful breeding. A new queen succeeded the old queen whilst she was still in olfactory contact. The time preceding queen succession was characterised by high levels of aggression, the death of a number of individuals, high cortisol and testosterone and low prolactin concentrations. Once the older queen was removed entirely and the new queen had given birth, prolactin concentrations increased and cortisol and testosterone concentrations decreased in subordinate NMRs. The results suggest that low prolactin levels are associated with low reproductive suppression during times of colony instability due to the removal or death of a queen.

Validating the use of dermal secretion as a matrix for monitoring glucocorticoid concentrations in African amphibian species.

The complex interaction between factors leading to amphibian declines is responsible for the inability to develop robust, standardized conservation action plans. Monitoring physiological stress responses in amphibians may provide an ideal tool to assist conservationists in this regard. This study aimed to validate dermal secretions as a robust matrix for monitoring glucocorticoid alterations in an African amphibian, the edible bullfrog (Pyxicephalus edulis). Both a biological (handling) and physiological (ACTH challenge) validation were conducted to determine which of five available enzyme immunoassays (EIAs) tested is most suited for monitoring alterations in dermal glucocorticoid (dGC) concentrations. Additionally, the most optimal body region for monitoring dGC concentrations in P. edulis was identified. To confirm the activation of the hypothalamic-pituitary-interrenal (HPI) axis following ACTH administration, urinary glucocorticoid metabolites (uGCM) were also quantified. The tested corticosterone EIA was the only assay able to monitor alterations in dGC concentrations following the handling event in P. edulis. Further validation during the ACTH challenge supported the corticosterone EIA in this regard. Dermal secretions collected from both the ventral and dorsal regions were suitable for monitoring dGC concentrations in both sexes. A considerable increase in uGCM concentrations following ACTH administration was found, suggesting that the dGC concentrations observed were derived from the HPI axis. This study offers further support for the use of dermal secretions as an important matrix for monitoring physiological stress, and thus general health, in amphibian species.

Analysis of gonadotrophin-releasing hormone-1 and kisspeptin neuronal systems in the nonphotoregulated seasonally breeding eastern rock elephant-shrew (Elephantulus myurus).

Of the 18 sub-Saharan elephant-shrew species, only eastern rock elephant-shrews reproduce seasonally throughout their distribution, a process seemingly independent of photoperiod. The present study characterizes gonadal status and location/intensity of gonadotrophin-releasing hormone-1 (GnRH-1) and kisspeptin immunoreactivities in this polyovulating species in the breeding and nonbreeding seasons. GnRH-1-immunoreactive (ir) cell bodies are predominantly in the medial septum, diagonal band, and medial preoptic area; processes are generally sparse except in the external median eminence. Kisspeptin-ir cell bodies are detected only within the arcuate nucleus; the density of processes is generally low, except in the septohypothalamic nucleus, ventromedial bed nucleus of the stria terminalis, arcuate nucleus, and internal and external median eminence. Kisspeptin-ir processes are negligible at locations containing GnRH-1-ir cell bodies. The external median eminence is the only site with conspicuously overlapping distributions of the respective immunoreactivities and, accordingly, a putative site for kisspeptin's regulation of GnRH-1 release in this species. In the nonbreeding season in males, there is an increase in the rostral population of GnRH-1-ir cell bodies and density of GnRH-1-ir processes in the median eminence. In both sexes, the breeding season is associated with increased kisspeptin-ir process density in the rostral periventricular area of the third ventricle and arcuate nucleus; at the latter site, this is positively correlated with gonadal mass. Cross-species comparisons lead us to hypothesize differential mechanisms within these peptidergic systems: that increased GnRH-1 immunoreactivity during the nonbreeding season reflects increased accumulation with reduced release; that increased kisspeptin immunoreactivity during the breeding season reflects increased synthesis with increased release.

Bottlenose dolphin (Tursiops truncatus) sperm revisited: Motility, morphology and ultrastructure of fresh sperm of consecutive ejaculates.

Computer aided sperm analysis systems allow detailed examination of sperm motility and morphology variables, which are important for the understanding of the spermatology of a species and the development of assisted reproductive techniques. Cetacean biology is too complex to study in the wild and data from captive individuals provide an important alternative for the conservation of these charismatic animals. The present study evaluates ejaculate and sperm characteristics, including sperm motility, kinematic variables and quantitative sperm morphology and ultrastructure, of consecutive ejaculates from Atlantic bottlenose dolphins (Tursiops truncatus). Sperm concentrations and total and progressive motilities were greater in the second than the first ejaculate, with all ejaculates being of very high quality (6.9-1127 × 106/ml sperm concentration, 75% to 91% total motility and 89% to 96% normal sperm). Most sperm in an ejaculate (≥84%) were highly (VCL>150 µm/s) and progressively motile with very few abnormal sperm. The sperm have small heads, a short but very bulky midpiece and a long tail. Detailed sperm morphometrics using CASA indicated there were similarities from one ejaculate to the next. The large mitochondria with extensive cristae mitochondriales are tightly packed in the midpiece resulting in a large midpiece volume. All the semen and sperm characteristics indicate high quality sperm and support the assumption that a multimale mating system is present in T. truncatus.

Non-invasive assessment of glucocorticoid and androgen metabolite levels in cooperatively breeding Damaraland mole-rats (Fukomys damarensis).

Dominant females of cooperative breeding species often use aggression to suppress reproduction of subordinate females, resulting in subordinates experiencing stress-related increases in glucocorticoid levels, which may cause reproductive down-regulation. This would suggest a general pattern with higher glucocorticoid levels in subordinate compared to dominant individuals; however, the opposite was found in a number of cooperatively breeding species. Furthermore, breeding females of the cooperatively breeding Damaraland mole-rats (Fukomys damarensis) exhibit very high androgen concentrations during the wet season, presumably to support their breeding monopoly. Hormone analysis in Damaraland mole-rats have typically been measured using plasma and urine, but faecal analysis offers additional advantages especially for field studies on this species. The present study examines the suitability of Damaraland mole-rat faecal samples for determining glucocorticoid metabolite (fGCM) and androgen metabolite (fAM) concentrations using enzyme immunoassays. Using these assays, we further evaluated the effects of breeding status on fGCM and fAM concentrations in wild-caught and captive Damaraland mole-rats. Wild-caught breeding and non-breeding males and females exhibited no differences in fAM concentrations. Immunoreactive fGCM concentrations were only high in male breeders and comparatively low in non-breeders and breeding females. Concentrations of fAMs and fGCMs were similar in captive males and females, but fAM concentrations were elevated in captive compared to wild-caught individuals, which may be related to a higher reproductive activity due to removal from the breeding female. The relatively uniform fAM and fGCM concentrations found in wild-caught mole-rats may be explained by a stable colony structure during the dry season during which this study was conducted. Limited dispersal opportunities result in lower aggression and stress levels within a colony and as a result lower fAM and fGCM concentrations.

Endoparasites of the Eastern Rock Sengi (Elephantulus myurus) from South Africa.

The endoparasite fauna of the eastern rock sengi ( Elephantulus myurus Thomas and Schwann) was studied for the first time for any sengi species from September 2007 until August 2008 in the Limpopo Province of South Africa. From the 121 sengis examined, we recovered 11 endoparasite taxa, including 9 nematodes, 1 cestode family (Hymenolepididae), and 1 pentastomid species (Armillifer armillatus (Wyman, 1834)). The overall endoparasite prevalence was high, at 100%, and largely attributable to the nematode Maupasina weissi Seurat, 1913 , with only a single individual being parasite free. Despite the high diversity, species richness was low (1.58 ± 0.06) and only M. weissi and spiruroid larvae occurred at a prevalence exceeding 8%. The abundance of M. weissi varied significantly between seasons and was lowest in summer and autumn. In contrast, the abundance of spiruroid larvae remained relatively constant across seasons in males, but was significantly higher in spring and summer compared to winter in females. These patterns may be generated by an accumulation of M. weissi with age as well as sex-specific seasonal shifts in diet. An updated list on the hosts and geographic range of parasites of sengis is provided.

The distribution of doublecortin-immunopositive cells in the brains of four afrotherian mammals: the Hottentot golden mole (Amblysomus hottentotus), the rock hyrax (Procavia capensis), the eastern rock sengi (Elephantulus myurus) and the four-toed sengi (Petrodromus tetradactylus).

Adult neurogenesis in the mammalian brain is now a widely accepted phenomenon, typically occurring in two forebrain structures: the subgranular zone (SGZ) of the hippocampal dentate gyrus and the subventricular zone (SVZ). Until recently, the majority of studies have focused on laboratory rodents, and it is under debate whether the process of adult neurogenesis occurs outside of the SGZ and the SVZ in other mammalian species. In the present study, we investigated potential adult neurogenetic sites in the brains of two elephant shrews/sengis, a golden mole and a rock hyrax, all members of the superorder Afrotheria. Doublecortin (DCX) immunoreactivity was used as a proxy to visualise adult neurogenesis, which is expressed in neuronal precursor cells and immature neurons. In all four species, densely packed DCX-positive cells were present in the SVZ, from where cells appear to migrate along the rostral migratory stream towards the olfactory bulb (OB). DCX-immunopositive cells were present in the granular cell layer and the glomerular layer of the OB. In the hippocampus, DCX-immunopositive cells were observed in the SGZ and in the granular layer of the dentate gyrus, with DCX-immunopositive processes extending into the molecular layer. In addition to these well-established adult neurogenic regions, DCX-immunopositive cells were also observed in layer II of the neocortex and the piriform cortex. While the present study reveals a similar pattern of adult neurogenesis to that reported previously in other mammals, further studies are needed to clarify if the cortical DCX-immunopositive cells are newly generated neurons or cells undergoing cortical remodelling.

Endoparasites of the spiny mouse (Acomys spinosissimus) from South Africa.

The endoparasite fauna of the spiny mouse (Acomys spinosissimus) was studied for the first time from April 2007 until April 2009 in a population from the Limpopo Province of South Africa. In a total of 129 mice examined, only 6 endoparasite taxa were found, 2 nematode species (Syphacia minuta, Monanema joopi), 1 genus of cestodes ( Rodentolepis spp.), and unidentified hymenolepidid fragments. In addition, 1 pentastomid species (Armillifer grandis) as well as unidentified porocephalid specimens were recovered. The overall prevalence was low, at 15.5%, and only 1 individual harbored more than 1 parasite species. With 12.4% prevalence, S. minuta was the most prevalent parasite. Its prevalence and abundance were significantly higher during the dry and cooler season than during the wet and warm season, while a female-biased burden was observed during the wet season only. For the remaining parasite species, low prevalence prevented meaningful statistical analyses. The observed parasite species richness, prevalence, and abundance for A. spinosissimus were low compared to values reported for other Acomys spp. This may be linked to the lack of anthropogenic influences in the study population as well as the small size of A. spinosissimus .

Eastern rock sengis as reservoir hosts of Anaplasma bovis in South Africa.

Recent work has identified the presence of Anaplasma bovis, a tick-borne pathogen of cattle, in a previously undescribed species of tick collected from eastern rock sengis, Elephantulus myurus, in Limpopo Province, South Africa. These small insectivores are endemic to Africa and are important hosts of immature ticks, however, their role as reservoirs of tick-borne pathogens has not been investigated. In order to elucidate the role of sengis in the epidemiology of A. bovis, we screened the blood of 105 sengis from Limpopo Province, South Africa, for the presence of members of the genera Anaplasma and Ehrlichia by PCR. A total of 30/105 (28.6%) of individuals were infected with A. bovis, and nucleotide sequencing revealed the presence of a novel genetic variant of this pathogen. This represents the first evidence that sengis may be natural reservoir hosts of A. bovis.

Natural hosts of the larvae of Nuttalliella sp. (N. namaqua?) (Acari: Nuttalliellidae).

The first collection of unengorged and fully engorged larvae of Nuttalliella sp. (N. namaqua?) from the murid rodents Micaelamys namaquensis, Aethomys chrysophilus and Acomys spinosissimus in Limpopo Province and from M. namaquensis in the Northern Cape Province, South Africa, is documented. A total of nine larvae were collected from two M. namaquensis in the Soutpansberg mountain range in the Limpopo Province during April 2009. During the last week of September 2011, 221 larvae were collected from rodents at the same locality and 10 of 48 M. namaquensis, 6 of 12 Ae. chrysophilus and 3 of 14 Ac. spinosissimus were infested. One of the M. namaquensis harboured 53 larvae. Five larvae were collected from two M. namaquensis in the Northern Cape Province. Total genomic DNA was extracted from two larvae and a region of the 18S rRNA gene was sequenced for these. BLASTn searches revealed similarity between these specimens and the Nuttalliella sequences published on GenBank.

Effects of life-history traits on parasitism in a monogamous mammal, the eastern rock sengi (Elephantulus myurus).

The distribution of parasites is often characterised by substantial aggregation with a small proportion of hosts harbouring the majority of parasites. This pattern can be generated by abiotic and biotic factors that affect hosts and determine host exposure and susceptibility to parasites. Climate factors can change a host's investment in life-history traits (e.g. growth, reproduction) generating temporal patterns of parasite aggregation. Similarly, host age may affect such investment. Furthermore, sex-biased parasitism is common among vertebrates and has been linked to sexual dimorphism in morphology, behaviour and physiology. Studies exploring sex-biased parasitism have been almost exclusively conducted on polygynous species where dimorphic traits are often correlated. We investigated the effects of season and life-history traits on tick loads of the monogamous eastern rock sengi (Elephantulus myurus). We found larger tick burdens during the non-breeding season possibly as a result of energetic constraints and/or climate effects on the tick. Reproductive investment resulted in increased larval abundance for females but not males and may be linked to sex-specific life-history strategies. The costs of reproduction could also explain the observed age effect with yearling individuals harbouring lower larval burdens than adults. Although adult males had the greatest larval tick loads, host sex appears to play a minor role in generating the observed parasite heterogeneities. Our study suggests that reproductive investment plays a major role for parasite patterns in the study species.

Jettisoning ballast or fuel? Caudal autotomy and locomotory energetics of the Cape dwarf gecko Lygodactylus capensis (Gekkonidae).

Many lizard species will shed their tail as a defensive response (e.g., to escape a putative predator or aggressive conspecific). This caudal autotomy incurs a number of costs as a result of loss of the tail itself, loss of resources (i.e., stored in the tail or due to the cost of regeneration), and altered behavior. Few studies have examined the metabolic costs of caudal autotomy. A previous study demonstrated that geckos can move faster after tail loss as a result of reduced weight or friction with the substrate; however, there are no data for the effects of caudal autotomy on locomotory energetics. We examined the effect of tail loss on locomotory costs in the Cape dwarf gecko Lygodactylus capensis ( approximately 0.9 g) using a novel method for collecting data on small lizards, a method previously used for arthropods. We measured CO(2) production during 5-10 min of exhaustive exercise (in response to stimulus) and during a 45-min recovery period. During exercise, we measured speed (for each meter moved) as well as total distance traveled. Contrary to our expectations, tailless geckos overall expended less effort in escape running, moving both slower and for a shorter distance, compared with when they were intact. Tailless geckos also exhibited lower excess CO(2) production (CO(2) production in excess of normal resting metabolic rate) during exercising. This may be due to reduced metabolically active tissue (tails represent 8.7% of their initial body mass). An alternative suggestion is that a change in energy substrate use may take place after tail loss. This is an intriguing finding that warrants future biochemical investigation before we can predict the relative costs of tail loss that lizards might experience under natural conditions.