Thermal biology in the Upper Galili Mountain blind mole rat (Nannospalax galili) and an overview of spalacine energetics.

Several hundred mammalian species thrive in complex burrow systems, which protect them from climatic extremes and predation. At the same time, it is also a stressful environment due to low food supply, high humidity, and, in some cases, a hypoxic and hypercapnic atmosphere. To face such conditions, subterranean rodents have convergently evolved low basal metabolic rate, high minimal thermal conductance and low body temperature. Although these parameters have been intensively studied in the last decades, such information is far from being well-known in one of the most studied groups of subterranean rodents, the blind mole rats of the genus Nannospalax. The lack of information is particularly noticeable for parameters such as the upper critical temperature and the width of the thermoneutral zone. In our study, we analysed the energetics of the Upper Galilee Mountain blind mole rat Nannospalax galili and found its basal metabolic rate of 0.84 ± 0.10 mL O2×g-1 × h-1, thermoneutral zone between 28 and 35 °C, mean Tb within the zone of 36.3 ± 0.6 °C, and minimal thermal conductance equal to 0.082 mL O2×g-1 × h-1 × C-1. Nannospalax galili is a truly homeothermic rodent well adapted to face lower ambient temperatures, because its Tb was stable down to the lowest temperature measured (10 °C). At the same time, a relatively high basal metabolic rate and relatively low minimal thermal conductance for a subterranean rodent of such body mass, and the difficulty of surviving ambient temperatures slightly above upper critical temperature, indicates problems with sufficient heat dissipation at higher temperatures. This can easily lead to overheating, that is relevant mainly during the hot-dry season. These findings suggest that N. galili can be threatened by ongoing global climate change.

Local Adaptation of Bitter Taste and Ecological Speciation in a Wild Mammal.

Sensory systems are attractive evolutionary models to address how organisms adapt to local environments that can cause ecological speciation. However, tests of these evolutionary models have focused on visual, auditory, and olfactory senses. Here, we show local adaptation of bitter taste receptor genes in two neighboring populations of a wild mammal-the blind mole rat Spalax galili-that show ecological speciation in divergent soil environments. We found that basalt-type bitter receptors showed higher response intensity and sensitivity compared with chalk-type ones using both genetic and cell-based functional analyses. Such functional changes could help animals adapted to basalt soil select plants with less bitterness from diverse local foods, whereas a weaker reception to bitter taste may allow consumption of a greater range of plants for animals inhabiting chalk soil with a scarcity of food supply. Our study shows divergent selection on food resources through local adaptation of bitter receptors, and suggests that taste plays an important yet underappreciated role in speciation.

Are southern African solitary mole-rats homeothermic or heterothermic under natural field conditions?

Abandoning of a stable body temperature (Tb), a phenomenon known as heterothermy, is an adaptation to cope mainly with a lack of food and water, especially in species inhabiting daily or seasonally variable environments. There is increasing evidence that African mammals avoid adverse conditions by heterothermy and eventually by entering torpor. Members of subterranean rodent family, the African mole-rats (Bathyergidae), are suitable candidates to study both phenomena, because of the diversity of their strategies in respect of maintaining stable Tb ranging from homeothermic species to a mammal with the most labile Tb, the naked mole-rat. Currently, there are field data on daily and seasonal Tb in one social species only and such information are lacking for any solitary mole-rat. In our study, we recorded yearly Tb in two solitary bathyergids, the Cape mole-rat Georychus capensis and the Cape dune mole-rat Bathyergus suillus from South Africa using intraperitoneally implanted dataloggers. Since this region is characterised by changing ecological characteristics, we expected either decreases of Tb within 24 h indicating daily torpor and/or longer-term decreases of Tb, which would indicate multiday torpor. Although we found seasonally phase shifted low amplitude daily Tb cycles, we did not find any remarkable and regular daily and/or seasonal Tb deviations, likely showing an absence of torpor in both species. Due to absence of this energy saving mechanism, we may speculate that both species could be vulnerable to ongoing global climatic change.

Heat dissipation in subterranean rodents: the role of body region and social organisation.

The relatively warm and very humid environment of burrows presents a challenge for thermoregulation of its mammalian inhabitants. It was found that African mole-rats dissipate body heat mainly through their venter, and social mole-rats dissipate more body heat compared to solitary species at lower temperatures. In addition, the pattern of the ventral surface temperature was suggested to be homogeneous in social mole-rats compared to a heterogeneous pattern in solitary mole-rats. To investigate this for subterranean rodents generally, we measured the surface temperatures of seven species with different degrees of sociality, phylogeny, and climate using infrared thermography. In all species, heat dissipation occurred mainly through the venter and the feet. Whereas the feet dissipated body heat at higher ambient temperatures and conserved it at lower ambient temperatures, the ventral surface temperature was relatively high in all temperatures indicating that heat dissipation to the environment through this body region is regulated mainly by behavioural means. Solitary species dissipated less heat through their dorsum than social species, and a tendency for this pattern was observed for the venter. The pattern of heterogeneity of surface temperature through the venter was not related to sociality of the various species. Our results demonstrate a general pattern of body heat exchange through the three studied body regions in subterranean rodents. Besides, isolated individuals of social species are less able to defend themselves against low ambient temperatures, which may handicap them if staying alone for a longer period, such as during and after dispersal events.

Genome evolution of blind subterranean mole rats: Adaptive peripatric versus sympatric speciation.

Speciation mechanisms remain controversial. Two speciation models occur in Israeli subterranean mole rats, genus Spalax: a regional speciation cline southward of four peripatric climatic chromosomal species and a local, geologic-edaphic, genic, and sympatric speciation. Here we highlight their genome evolution. The five species were separated into five genetic clusters by single nucleotide polymorphisms, copy number variations (CNVs), repeatome, and methylome in sympatry. The regional interspecific divergence correspond to Pleistocene climatic cycles. Climate warmings caused chromosomal speciation. Triple effective population size, Ne , declines match glacial cold cycles. Adaptive genes evolved under positive selection to underground stresses and to divergent climates, involving interspecies reproductive isolation. Genomic islands evolved mainly due to adaptive evolution involving ancient polymorphisms. Repeatome, including both CNV and LINE1 repetitive elements, separated the five species. Methylation in sympatry identified geologically chalk-basalt species that differentially affect thermoregulation, hypoxia, DNA repair, P53, and other pathways. Genome adaptive evolution highlights climatic and geologic-edaphic stress evolution and the two speciation models, peripatric and sympatric.

Gas composition and its daily changes within burrows and nests of an Afroalpine fossorial rodent, the giant root-rat Tachyoryctes macrocephalus.

Fossorial mammals are supposed to face hypoxic and hypercapnic conditions, but such conditions have been rarely encountered in their natural burrow systems. Gas composition in burrows after heavy rains, deeper burrows and especially nest chambers, where animals usually spend most of the day, could be even more challenging than in shallow burrows. Such situations, however, have been rarely surveyed in the wild. In our study, we determined concentrations of O2, CO2 and CH4 in active burrows and nests of the giant root-rat Tachyoryctes macrocephalus, a large fossorial rodent endemic to the Afroalpine zone of the Bale Mountains in Ethiopia. We were able to determine the precise location of nests by tracking individuals equipped with radio-collars. To the best of our knowledge, this is the first study that analyses air samples taken directly from the nests of actually occupied burrow systems in any free-living fossorial mammal. We found no evidence for environmental hypoxia in the examined burrows and nests (range 19.7-21.6% O2). Concentrations of CO2 in the burrows increased after the burrows were plugged in the evening, but did not reach physiologically problematic levels. The highest CO2 concentrations in burrows were detected in the evening during a wet period (up to 0.44%). In root-rat nest chambers, the highest (but still harmless) CO2 concentrations (up to 1.31%) were detected in the morning (measured in the late dry season only) together with an elevated concentration (up to 13.5ppm) of CH4. Regular surface activity of giant root-rats, combined with the relatively large dimensions of their nest chambers and tunnels, and the absence of heavy soils, may contribute to harmless atmospheres within their burrow systems.

Ecological role of the giant root-rat (Tachyoryctes macrocephalus) in the Afroalpine ecosystem.

Rodents with prevailing subterranean activity usually play an important role in the ecosystems of which they are a part due to the combined effect of herbivory and soil perturbation. This is the case for the giant root-rat Tachyoryctes macrocephalus endemic to the Afroalpine ecosystem of the Bale Mountains, Ethiopia. We studied the impact of root-rats on various ecosystem features within a 3.5-ha study locality dominated by Alchemilla pasture, which represents an optimal habitat for this species, in 2 periods of a year. The root-rats altered plant species composition, reducing the dominant forb, Alchemilla abyssinica, while enhancing Salvia merjame and a few other species, and reduced vegetation cover, but not the fresh plant biomass. Where burrows were abandoned by root-rats, other rodents took them over and A. abyssinica increased again. Root-rat burrowing created small-scale heterogeneity in soil compactness due to the backfilling of some unused burrow segments. Less compacted soil tended to be rich in nutrients, including carbon, nitrogen and phosphorus, which likely affected the plant growth on sites where the vegetation has been reduced as a result of root-rat foraging and burrowing.

Variability of space-use patterns in a free living eusocial rodent, Ansell's mole-rat indicates age-based rather than caste polyethism.

Eusocial species of African mole-rats live in groups cooperating on multiple tasks and employing division of labour. In captivity, individuals of the same group differ in cooperative contribution as well as in preference for a particular task. Both can be viewed as polyethism. However, little information is available from free-ranging mole-rats, which live in large burrow systems. We made an attempt to detect polyethism in the free-living Ansell's mole-rat (Fukomys anselli) as differences in individuals' space-use patterns. We radio-tracked 17 adults from five groups. Large individuals, including breeding males, spent more time inside the nest than smaller individuals. Breeding females were more often located <10 m from the nest in comparison to non-breeding females, who were relatively more often located 30-90 m and exclusively >90 m from the nest. One non-breeding female even conducted a brief intrusion into a neighbouring group's territory via an open tunnel connection. A significant part of the variability in mole-rat space-use patterns was explained by body mass which is probably related to age in this species. This result can therefore be attributed to age polyethism. There was no apparent discontinuity in the space-use patterns of non-breeders that would indicate existence of castes.

Sympatric speciation of spiny mice, Acomys, unfolded transcriptomically at Evolution Canyon, Israel.

Spiny mice, Acomys cahirinus, colonized Israel 30,000 y ago from dry tropical Africa and inhabited rocky habitats across Israel. Earlier, we had shown by mtDNA that A. cahirinus incipiently sympatrically speciates at Evolution Canyon I (EC I) in Mount Carmel, Israel because of microclimatic interslope divergence. The EC I microsite consists of a dry and hot savannoid "African" slope (AS) and an abutting humid and cool-forested "European" slope (ES). Here, we substantiate incipient SS in A. cahirinus at EC I based on the entire transcriptome, showing that multiple slope-specific adaptive complexes across the transcriptome result in two divergent clusters. Tajima's D distribution of the abutting Acomys interslope populations shows that the ES population is under stronger positive selection, whereas the AS population is under balancing selection, harboring higher genetic polymorphisms. Considerable sites of the two populations were differentiated with a coefficient of FST = 0.25-0.75. Remarkably, 24 and 37 putatively adaptively selected genes were detected in the AS and ES populations, respectively. The AS genes involved DNA repair, growth arrest, neural cell differentiation, and heat-shock proteins adapting to the local AS stresses of high solar radiation, drought, and high temperature. In contrast, the ES genes involved high ATP associated with energetics stress. The sharp ecological interslope divergence led to strong slope-specific selection overruling the interslope gene flow. Earlier tests suggested slope-specific mate choice. Habitat interslope-adaptive selection across the transcriptome and mate choice substantiate sympatric speciation (SS), suggesting its prevalence at EC I and commonality in nature.

Transcriptome, genetic editing, and microRNA divergence substantiate sympatric speciation of blind mole rat, Spalax.

Incipient sympatric speciation in blind mole rat, Spalax galili, in Israel, caused by sharp ecological divergence of abutting chalk-basalt ecologies, has been proposed previously based on mitochondrial and whole-genome nuclear DNA. Here, we present new evidence, including transcriptome, DNA editing, microRNA, and codon usage, substantiating earlier evidence for adaptive divergence in the abutting chalk and basalt populations. Genetic divergence, based on the previous and new evidence, is ongoing despite restricted gene flow between the two populations. The principal component analysis, neighbor-joining tree, and genetic structure analysis of the transcriptome clearly show the clustered divergent two mole rat populations. Gene-expression level analysis indicates that the population transcriptome divergence is displayed not only by soil divergence but also by sex. Gene ontology enrichment of the differentially expressed genes from the two abutting soil populations highlights reproductive isolation. Alternative splicing variation of the two abutting soil populations displays two distinct splicing patterns. L-shaped FST distribution indicates that the two populations have undergone divergence with gene flow. Transcriptome divergent genes highlight neurogenetics and nutrition characterizing the chalk population, and energetics, metabolism, musculature, and sensory perception characterizing the abutting basalt population. Remarkably, microRNAs also display divergence between the two populations. The GC content is significantly higher in chalk than in basalt, and stress-response genes mostly prefer nonoptimal codons. The multiple lines of evidence of ecological-genomic and genetic divergence highlight that natural selection overrules the gene flow between the two abutting populations, substantiating the sharp ecological chalk-basalt divergence driving sympatric speciation.

Habitat and Burrow System Characteristics of the Blind Mole Rat Spalax galili in an Area of Supposed Sympatric Speciation.

A costly search for food in subterranean rodents resulted in various adaptations improving their foraging success under given ecological conditions. In Spalax ehrenbergi superspecies, adaptations to local ecological conditions can promote speciation, which was recently supposed to occur even in sympatry at sites where two soil types of contrasting characteristics abut each other. Quantitative description of ecological conditions in such a site has been, nevertheless, missing. We measured characteristics of food supply and soil within 16 home ranges of blind mole rats Spalax galili in an area subdivided into two parts formed by basaltic soil and pale rendzina. We also mapped nine complete mole rat burrow systems to compare burrowing patterns between the soil types. Basaltic soil had a higher food supply and was harder than rendzina even under higher moisture content and lower bulk density. Population density of mole rats was five-times lower in rendzina, possibly due to the lower food supply and higher cover of Sarcopoterium shrubs which seem to be avoided by mole rats. A combination of food supply and soil parameters probably influences burrowing patterns resulting in shorter and more complex burrow systems in basaltic soil.

Social and Environmental Influences on Daily Activity Pattern in Free-Living Subterranean Rodents: The Case of a Eusocial Bathyergid.

Predictable daily activity patterns have been detected repeatedly even in mammals living in stable environments, as is the case for subterranean rodents. Whereas studies on activity of these rodents under laboratory conditions almost exclusively have concerned themselves with the influence of light, many field studies have revealed signs of an association between the activity pattern and daily fluctuations of temperature under the ground. This would assume that behavioral thermoregulation is probably involved. The only exceptions to the relationship between temperature and activity are 2 eusocial mole-rats of the genus Fukomys (Bathyergidae, Rodentia), which indicates that activity patterns could be affected also by social cues. To better understand how social and environmental factors influence the activity pattern in a eusocial mole-rat, we monitored the outside-nest activity in another species of this genus, the Ansell's mole-rat (Fukomys anselli), which has a relatively small body mass, high conductance, and more superficially situated burrows. Its daily activity had 1 prominent peak (around 1400 h), and it was tightly correlated with the temperature measured at depth of foraging burrows. Since F. anselli has high thermoregulatory requirements to maintain stable body temperature below the lower critical temperature, we conclude that the observed pattern is probably the result of minimizing the cost of thermoregulation. There were no significant differences in the daily activity patterns of breeding males and females and nonbreeders. Members of the same family group tended to have more similar activity patterns, but consistent activity synchronization between individuals was not proven. From the comparison of available data on all subterranean rodents, we assume that social cues in communally nesting mole-rats may disrupt (mask) temperature-related daily activity rhythms but probably only if the additional cost of thermoregulation is not too high, as it likely is in the Ansell's mole-rat.

Spatial and temporal activity patterns of the free-living giant mole-rat (Fukomys mechowii), the largest social bathyergid.

Despite the considerable attention devoted to the biology of social species of African mole-rats (Bathyergidae, Rodentia), knowledge is lacking about their behaviour under natural conditions. We studied activity of the largest social bathyergid, the giant mole-rat Fukomys mechowii, in its natural habitat in Zambia using radio-telemetry. We radio-tracked six individuals during three continuous 72-h sessions. Five of these individuals, including a breeding male, belonged to a single family group; the remaining female was probably a solitary disperser. The non-breeders of the family were active (i.e. outside the nest) 5.8 hours per 24h-day with the activity split into 6.5 short bouts. The activity was more concentrated in the night hours, when the animals also travelled longer distances from the nest. The breeding male spent only 3.2 hours per day outside the nest, utilizing less than 20% of the whole family home range. The dispersing female displayed a much different activity pattern than the family members. Her 8.0 hours of outside-nest activity per day were split into 4.6 bouts which were twice as long as in the family non-breeders. Her activity peak in the late afternoon coincided with the temperature maximum in the depth of 10 cm (roughly the depth of the foraging tunnels). Our results suggest that the breeding individuals (at least males) contribute very little to the work of the family group. Nevertheless, the amount of an individual's activity and its daily pattern are probably flexible in this species and can be modified in response to actual environmental and social conditions.

Possible incipient sympatric ecological speciation in blind mole rats (Spalax).

Sympatric speciation has been controversial since it was first proposed as a mode of speciation. Subterranean blind mole rats (Spalacidae) are considered to speciate allopatrically or peripatrically. Here, we report a possible incipient sympatric adaptive ecological speciation in Spalax galili (2n = 52). The study microsite (0.04 km(2)) is sharply subdivided geologically, edaphically, and ecologically into abutting barrier-free ecologies divergent in rock, soil, and vegetation types. The Pleistocene Alma basalt abuts the Cretaceous Senonian Kerem Ben Zimra chalk. Only 28% of 112 plant species were shared between the soils. We examined mitochondrial DNA in the control region and ATP6 in 28 mole rats from basalt and in 14 from chalk habitats. We also sequenced the complete mtDNA (16,423 bp) of four animals, two from each soil type. Remarkably, the frequency of all major haplotype clusters (HC) was highly soil-biased. HCI and HCII are chalk biased. HC-III was abundant in basalt (36%) but absent in chalk; HC-IV was prevalent in basalt (46.5%) but was low (20%) in chalk. Up to 40% of the mtDNA diversity was edaphically dependent, suggesting constrained gene flow. We identified a homologous recombinant mtDNA in the basalt/chalk studied area. Phenotypically significant divergences differentiate the two populations, inhabiting different soils, in adaptive oxygen consumption and in the amount of outside-nest activity. This identification of a possible incipient sympatric adaptive ecological speciation caused by natural selection indirectly refutes the allopatric alternative. Sympatric ecological speciation may be more prevalent in nature because of abundant and sharply abutting divergent ecologies.

A seasonal difference of daily energy expenditure in a free-living subterranean rodent, the silvery mole-rat (Heliophobius argenteocinereus; Bathyergidae).

In seasonal climatic regimes, animals have to deal with changing environmental conditions. It is reasonable to expect that seasonal changes are reflected in animal overall energetics. The relation between daily energy expenditure (DEE) and seasonally variable ecological determinants has been studied in many free-living small mammals; however with inconsistent results. Subterranean mammals, i.e. fossorial (burrowing) mammals which live and forage underground, live in a seasonally and diurnally thermally stable environment and represent a suitable model to test seasonality in DEE in respect to seasonal changes, particularly those in soil characteristics and access to food supply. Both factors are affected by seasonal rainfall and are supposed to fundamentally determine activity of belowground dwellers. These ecological constraints are pronounced in some tropical regions, where two distinct periods, dry and rainy seasons, regularly alternate. To explore how a tropical mammal responds to an abrupt environmental change, we determined DEE, resting metabolic rate (RMR) and sustained metabolic scope (SusMS) in a solitary subterranean rodent, the silvery mole-rat, at the end of dry season and the onset of rainy season. Whereas RMR did not differ between both periods, mole-rats had 1.4 times higher DEE and SusMS after the first heavy rains. These findings suggest that rainfall is an important environmental factor responsible for higher energy expenditure in mole-rats, probably due to increased burrowing activity. SusMS in the silvery mole-rat is comparable to values in other bathyergids and all bathyergid values rank among the lowest SusMS found in endothermic vertebrates.