Regional Intraspecific Differences of Thermal Biology in a Marsupial Hibernator.

AbstractDuring periods of torpor, hibernators can reduce metabolic rate (MR) and body temperature (Tb) substantially. However, to avoid physiological dysfunction at low temperatures, they defend Tb at a critical minimum, often between ~0°C and 10°C via an increase in MR. Because thermoregulation during torpor requires extra energy, individuals with lower Tb's and thus minimal MR during torpor should be selected in colder climates. Such inter- and intraspecific variations occur in some placental mammals, but for the evolutionary separate marsupials, available information is scarce. Marsupial eastern pygmy possums (Cercartetus nanus; ~22 g body mass), widely distributed along the Australian southeastern coast including subtropical to alpine areas, were used to test the hypothesis that the defended Tb of torpid individuals is related to the climate of their habitat. Possums were captured from five regions, 1,515 km apart, with midwinter (July) minimum environmental temperatures (min Tenv's) ranging from -3.9°C to 6.6°C. Captive possums in deep torpor were slowly cooled with ambient temperature (Ta), while their MR was measured to determine the minimum torpor metabolic rate (TMR), the Ta at which their MR increased for thermoregulation (min Ta), and the corresponding minimum Tb (min Tb). Partial least squares regression analysis revealed that Ta and Tenv were the strongest explanatory variables for the min Tb. The min Tb and Ta were also correlated with latitude but not elevation of the capture sites. However, the best correlations were observed between the min Tenv and the min Tb and Ta for individuals experiencing min Tenv>0°C; these individuals thermoconformed to min Ta's between -0.8°C and 3.7°C, and their min Tb ranged from 0.5°C to 6.0°C and was 0.5°C-2.6°C below the min Tenv at the capture site. In contrast, individuals experiencing a min Tenv of -3.9°C regulated Tb at 0.6°C±0.2°C or 4.5°C above the Tenv. The minimum TMR of all possums did not differ with Ta and thus did not differ among populations and was 2.6% of the basal MR. These data provide new evidence that thermal variables of marsupials are subject to regional intraspecific variation. It suggests that min Tb is a function of the min Tenv but only above 0°C, perhaps because the Tb-Ta differential for torpid possums in the wild, at a min Tenv of -3.9°C, remains small enough to be compensated by a small increase in MR and does not require the physiological capability for a reduction of Tb below 0°C.

Two-stage evolution of mammalian adipose tissue thermogenesis.

Brown adipose tissue (BAT) is a heater organ that expresses thermogenic uncoupling protein 1 (UCP1) to maintain high body temperatures during cold stress. BAT thermogenesis is considered an overarching mammalian trait, but its evolutionary origin is unknown. We show that adipose tissue of marsupials, which diverged from eutherian mammals ~150 million years ago, expresses a nonthermogenic UCP1 variant governed by a partial transcriptomic BAT signature similar to that found in eutherian beige adipose tissue. We found that the reconstructed UCP1 sequence of the common eutherian ancestor displayed typical thermogenic activity, whereas therian ancestor UCP1 is nonthermogenic. Thus, mammalian adipose tissue thermogenesis may have evolved in two distinct stages, with a prethermogenic stage in the common therian ancestor linking UCP1 expression to adipose tissue and thermal stress. We propose that in a second stage, UCP1 acquired its thermogenic function specifically in eutherians, such that the onset of mammalian BAT thermogenesis occurred only after the divergence from marsupials.

Climate change and population persistence in a hibernating marsupial.

Climate change has physiological consequences on organisms, ecosystems and human societies, surpassing the pace of organismal adaptation. Hibernating mammals are particularly vulnerable as winter survival is determined by short-term physiological changes triggered by temperature. In these animals, winter temperatures cannot surpass a certain threshold, above which hibernators arouse from torpor, increasing several fold their energy needs when food is unavailable. Here, we parameterized a numerical model predicting energy consumption in heterothermic species and modelled winter survival at different climate change scenarios. As a model species, we used the arboreal marsupial monito del monte (genus Dromiciops), which is recognized as one of the few South American hibernators. We modelled four climate change scenarios (from optimistic to pessimistic) based on IPCC projections, predicting that northern and coastal populations (Dromiciops bozinovici) will decline because the minimum number of cold days needed to survive the winter will not be attained. These populations are also the most affected by habitat fragmentation and changes in land use. Conversely, Andean and other highland populations, in cooler environments, are predicted to persist and thrive. Given the widespread presence of hibernating mammals around the world, models based on simple physiological parameters, such as this one, are becoming essential for predicting species responses to warming in the short term.

Fear of the human 'super predator' in native marsupials and introduced deer in Australia.

Recent experiments have demonstrated that carnivores and ungulates in Africa, Asia, Europe and North America fear the human 'super predator' far more than other predators. Australian mammals have been a focus of research on predator naiveté because it is suspected they show atypical antipredator responses. To experimentally test if mammals in Australia also most fear humans, we quantified the responses of four native marsupials (eastern grey kangaroo, Bennett's wallaby, Tasmanian pademelon, common brushtail possum) and introduced fallow deer to playbacks of predator (human, dog, Tasmanian devil, wolf) or non-predator control (sheep) vocalizations. Native marsupials most feared the human 'super predator', fleeing humans 2.4 times more often than the next most frightening predator (dogs), and being most, and significantly, vigilant to humans. These results demonstrate that native marsupials are not naïve to the peril humans pose, substantially expanding the taxonomic and geographic scope of the growing experimental evidence that wildlife worldwide generally perceive humans as the planet's most frightening predator. Introduced fallow deer fled humans, but not more than other predators, which we suggest may result from their being introduced. Our results point to both challenges concerning marsupial conservation and opportunities for exploiting fear of humans as a wildlife management tool.

Semelparous marsupials reduce sleep for sex.

Sleep is a prominent, seemingly universal animal behavior. Although sleep maintains optimal waking performance, the biological drive to sleep may be incompatible with the life history of some species. In a multi-year study on semelparous marsupials in Australia, we provide the first direct evidence of ecological sleep restriction in a terrestrial mammal. Dusky (Antechinus swainsonii) and agile (A. agilis) antechinus have an unusual reproductive strategy characterized by the synchronous death of all males at the end of their only breeding season. Using accelerometry, electrophysiology, and metabolomics, we show that males, but not females, increase their activity during the breeding season by reducing sleep. In a trade-off between the neurophysiological requirements for sleep and evolutionary necessity for reproduction, strong sexual selection might drive males to sacrifice sleep to increase access to fertile females and ultimately maximize their fitness.

Sugar Glider Pediatrics.

Sugar gliders (Petaurus breviceps) are small nocturnal marsupials with a unique metabolism and reproductive strategy compared to other common exotic companion mammals. Although there are few reports on the pediatric diseases of sugar gliders, clinicians should be aware of the normal reproductive anatomy, physiology, and rearing of sugar gliders. By recognizing the signs of normal reproduction and joey development, clinicians can identify abnormalities quickly and provide appropriate intervention, as needed.

Over-eruption in marsupial carnivore teeth: compensation for a constraint.

Pronounced over-eruption of the canine teeth, causing the cervical enamel margin to extend beyond the alveolar bone and exposing the root, occurs with age and growth in Australian marsupial carnivores, much more than in eco-morphologically equivalent placental carnivores. Suppression of functional tooth replacement is characteristic of marsupials, where most placentals have the primitive diphyodont pattern of two generations of incisor, canine and premolar teeth. Canine and molar tooth dimensions of four species of marsupial carnivores (thylacine Thylacinus cynocephalus, Tasmanian devil Sarcophilus harrisii and two quolls Dasyurus spp.) and canine dimensions of seven eco-morphologically equivalent placental carnivore species were measured from museum specimens. Canine dimensions were measured in a time series on live wild-living individual devils and quolls. The canine teeth and to a lesser extent the molar teeth of marsupial carnivores continue to erupt through life, resulting in a net increase in tooth height and diameter, a phenomenon not evident in placental carnivores. Potential mechanisms causing over-eruption include tooth wear and gradual release of occlusal pressure as the individual grows. Over-eruption in marsupial carnivores may be a compensatory response for tooth size limits imposed by monophyodont tooth replacement, ensuring that animal's teeth are scaled to jaw size from juvenile to adulthood.

Long-term survival, temperature, and torpor patterns.

Mammalian and avian torpor is highly effective in reducing energy expenditure. However, the extent of energy savings achieved and thus long-term survival appear to differ between species capable of multiday hibernation and species restricted to daily heterothermy, which could, however, be due to thermal effects. We tested how long-term survival on stored body fat (i.e. time to lean body mass), crucial for overcoming adverse periods, is related to the pattern of torpor expressed under different ambient temperatures (Ta: 7 °C typical of hibernation, 15 and 22 °C typical of daily torpor) in the small marsupial hibernator the pygmy-possum (Cercartetus nanus). Possums expressed torpor at all Tas and survived without food for 310 days on average at Ta 7 °C, 195 days at Ta 15 °C, and 127 days at Ta 22 °C. At Ta 7 and 15 °C, torpor bout duration (TBD) increased from < 1-3 to ~ 5-16 days over 2 months, whereas at Ta 22 °C, TBD remained at < 1 to ~ 2 days. At all Tas daily energy use was substantially lower and TBD and survival times of possums much longer (3-12 months) than in daily heterotherms (~ 10 days). Such pronounced differences in torpor patterns and survival times even under similar thermal conditions provide strong support for the concept that torpor in hibernators and daily heterotherms are physiologically distinct and have evolved for different ecological purposes.

Flexible Employment of Torpor in Squirrel Gliders (Petaurus norfolcensis): An Adaptation to Unpredictable Climate?

AbstractTorpor is a highly effective response to counter various ecological and physiological bottlenecks in endotherms. In this study, we examined interrelations between thermoregulatory responses and key environmental variables in free-living squirrel gliders (Petaurus norfolcensis) in a habitat with drastic climatic and ecological changes across seasons. To this end, we measured body temperature (Tb) and heart rate (fH) simultaneously throughout the year using implanted data loggers. Squirrel gliders in our study experienced fluctuations in ambient temperature (Ta) between -4.0°C and 44.1°C and expressed torpor at different times during the year. In contrast to our expectations, torpor seemed to be employed flexibly, on demand, and most frequently in spring rather than during the coldest and/or hottest periods. Torpor bouts lasted, on average, about 5 h, and Tb during torpor dropped as low as 17.9°C. The fH during torpor decreased below 50 bpm, which is about one-third of the basal level. The ability to record fH alongside Tb enabled us to also report periods of low fH during thermoconforming hyperthermia at Ta's above 35°C that likely occurred to conserve energy and water. Our findings double the body size of Australian gliders for which data on torpor are available and advance our ecological understanding of the dynamics of torpor expression in wild mammals and of how animals cope with varying conditions. Moreover, they highlight that the flexibility of physiology and thermoregulatory responses are clearly more complex than previously thought.

Communal nesting is the optimal strategy for heat conservation in a social marsupial: lessons from biophysical models.

Endothermy, understood as the maintenance of continuous and high body temperatures owing to the combination of metabolic heat production and an insulative cover, is severely challenged in small endotherms inhabiting cold environments. As a response, social clustering combined with nest use (=communal nesting) is a common strategy for heat conservation. To quantify the actual amount of energy that is saved by this strategy, we studied the social marsupial Dromiciops gliroides (monito del monte), an endemic species of the cold forests of southern South America. It is hypothesized that sociability in this marsupial was driven by cold conditions, but evidence supporting this hypothesis is unclear. Here, we used taxidermic models ('mannequins') to experimentally test the energetic benefits of clustering combined with nest use. To do this, we fitted and compared cooling curves of solitary and grouped mannequins, within and outside of a nest, at the typical winter ambient temperatures of their habitat (5°C). We found that the strategy that minimized euthermic cost of maintenance was the combination of nest use and clustering, thus supporting communal nesting as a social adaptation to cope with the cold. Considering the basal metabolic rate of monitos, our estimates suggest that the savings represents almost half of energy consumption per day (in resting conditions). This study shows how simple biophysical models could help to evaluate bioenergetic hypotheses for social behavior in cold-adapted endotherms.

Sensitivity of the stripe-faced dunnart, Sminthopsis macroura (Gould 1845), to the insecticide, fipronil; implications for pesticide risk assessments in Australia.

A lack of toxicity data quantifying responses of Australian native mammals to agricultural pesticides prompted an investigation into the sensitivity of the stripe-faced dunnart, Sminthopsis macroura (Gould 1845) to the insecticide, fipronil (5-amino-3-cyano-1-(2,6-dichloro-4-trifluoromethylphenyl)-4-trifluoromethylsulfinyl pyrazole, CAS No. 120068-37-3). Using the Up-And-Down method for determining acute oral toxicity in mammals (OECD) median lethal dose estimates of 990 mg kg-1 (95% confidence interval (CI) = 580.7-4770.0 mg kg-1) and 270.4 mg kg-1 (95% CI = 0.0->20,000.0 mg kg-1) were resolved for male and female S. macroura, respectively. The difference between median lethal dose estimates for males and females may have been influenced by the older ages of two female dunnarts. Consequently, further modelling of female responses to fipronil doses used the following assumptions: (a) death at 2000 mg kg-1, (b) survival at 500 mg kg-1 and (c) a differential response (both survival and death) at 990 mg kg-1. This modelling revealed median lethal dose estimates for female S. macroura of 669.1 mg kg-1 (95% CI = 550-990 mg kg-1; assuming death at 990 mg kg-1) and 990 mg kg-1 (95% CI = 544.7-1470 mg kg-1; assuming survival at 990 mg kg-1). These median lethal dose estimates are 3-10-fold higher than available LD50 values of 94 mg kg-1 for a similarly sized eutherian mammal, Mus musculus (L. 1758) and 97 mg kg-1 for Rattus norvegicus (Birkenhout 1769). Implications for pesticide risk assessments in Australia are discussed.

Small Alpine Marsupials Regulate Evaporative Water Loss, Suggesting a Thermoregulatory Role Rather than a Water Conservation Role.

AbstractWe show here that evaporative water loss (EWL) is constant over a wide range of ambient relative humidity for two species of small, mesic habitat dasyurid marsupials (Antechinus agilis and Antechinus swainsonii) below thermoneutrality (20°C) and within thermoneutrality (30°C). This independence of EWL from the water vapor pressure deficit between the animal and its environment indicates that EWL is physiologically controlled by both species. The magnitude of this control of EWL was similar to that of two other small marsupials from more arid habitats, which combined with the observation that there were no effects of relative humidity on body temperature or metabolic rate, suggests that control of EWL is a consequence of precise thermoregulation to maintain heat balance rather than a water-conserving strategy at low relative humidities. The antechinus appear to manipulate cutaneous EWL rather than respiratory EWL to control their total EWL by modifying their cutaneous resistance and/or skin temperature. We propose that there is a continuum between enhanced thermoregulatory EWL at high ambient temperature and so-called insensible EWL at and below thermoneutrality.

Running of the feathertail glider (Acrobates pygmaeus) on level ground: Kinematics.

A complete kinematic analysis of the trunk and limbs was performed for the smallest gliding mammal, the feathertail glider (Acrobates pygmaeus). To compare the running technique of the glider with that of terrestrial mammals, only movement on horizontal substrate was studied. Kinematic analysis was performed separately for bound with extended suspension and bound with both suspensions. Terrestrial locomotion of the feathertail glider was classified as primitive ricochet, which is distinguished from true gallop by the absence of a delay in protraction of the hindlimbs after their take-off. The kinematic analysis showed significant impacts of gliding adaptation on the quadrupedal running, the most striking of which are the extremely sprawling position of the limbs, in particular, the wide stance of the forelimbs, and consequently, the absence of crossing of the fore- and hindlimbs in gathered stage of the cycle. The degree of limbs' sprawling in the feathertail glider is closer to the reptilian condition than to the mammalian parasagittal state. The sprawling condition of the feathertail glider is secondary and therefore is designated as "deparasagittalization."

Running of the feathertail glider (Acrobates pygmaeus) on level ground: Gaits.

Gliding is a crucial adaptation to arboreal habitats in several groups of mammals. Along with certain advantages it imposes limitations on the quadrupedal running since it affects the locomotor apparatus. To estimate the impact on quadrupedal running in gliders, the feathertail glider (Acrobates pygmaeus) was chosen considering that the small size allows minor morphological modifications for aerial locomotion. The gaits were studied on horizontal flat substrate which made it possible to compare running technique of the glider with that of terrestrial mammals. In all analyzed plots the footfall sequence was found to be asymmetrical; in most cases the bound was used, in contrast, the gallop occurred only occasionally. The half-bound with the fore lead, the most usual asymmetrical gait in quadrupedal marsupials, was much less common in A. pygmaeus than the bound; the rare among mammals half-bound with the hind lead was also found. The bound was not only the most common gait but also the steadiest one; therefore we can conclude that A. pygmaeus uses all other asymmetrical gaits as transitional forms associated with changes in speed, direction, etc. The bound with extended suspension is probably preferred by A. pygmaeus because it most closely resembles gliding by posture.

Passive acoustic monitoring for detecting the Yellow-bellied Glider, a highly vocal arboreal marsupial.

Passive acoustic monitoring (PAM) is increasingly being used for the survey of vocalising wildlife species that are otherwise cryptic and difficult to survey. Our study aimed to develop PAM guidelines for detecting the Yellow-bellied Glider, a highly vocal arboreal marsupial that occurs in native Eucalyptus forests in eastern and south-eastern Australia. To achieve this, we considered the influence of background noise, weather conditions, lunar illumination, time since sunset and season on the probability of detecting vocalisations. We deployed Autonomous Recording Units (ARUs) at 43 sites in the Central Highlands of Victoria during two periods: spring/summer (October 2018 to January 2019), and autumn/winter (May to August 2019). ARUs were programmed to record for 11 hours from sunset for 14 consecutive days during each period. Background noise resulted from inclement weather (wind and rain) and masked vocalisations in spectrograms of the recordings, thus having the greatest influence on detection probability. Vocalisations were most common in the four hours after sunset. Rainfall negatively influenced detection probability, especially during the autumn/winter sampling period. Detection of Yellow-bellied Gliders with PAM requires deploying ARUs programmed to record for four hours after sunset, for a minimum of six nights with minimal inclement weather (light or no wind or rain). The survey period should be extended to 12 nights when rain or wind are forecast. Because PAM is less labour intensive than active surveys (i.e., spotlighting and call playbacks with multiple observers and several nights' survey per site), its use will facilitate broad-scale surveys for Yellow-bellied Gliders.

Pros and cons for the evidence of adaptive non-shivering thermogenesis in marsupials.

The thermogenic mechanisms supporting endothermy are still not fully understood in all major mammalian subgroups. In placental mammals, brown adipose tissue currently represents the most accepted source of adaptive non-shivering thermogenesis. Its mitochondrial protein UCP1 (uncoupling protein 1) catalyzes heat production, but the conservation of this mechanism is unclear in non-placental mammals and lost in some placentals. Here, we review the evidence for and against adaptive non-shivering thermogenesis in marsupials, which diverged from placentals about 120-160 million years ago. We critically discuss potential mechanisms that may be involved in the heat-generating process among marsupials.

Red-tailed phascogales: A review of their biology and importance as model marsupial species.

There are many limitations when using traditional laboratory species. Limits on variation, may result in limited outcomes, at both the species and individual level, due to different individuals/species having diverse physiological processes, or differing molecular and genetic mechanisms. By using a variety of model species, we will be able to develop creative solutions to biological problems and identify differences of which we were not previously aware. The laboratory mouse has been a suitable model species for various mammalian studies, however most are bred specifically for laboratory research with limited variability due to selective breeding. Marsupial models offer unique research opportunities compared to eutherian models. We believe that there should be an expansion in marsupial model species, and the introduction of the red-tailed phascogale (Phascogale calura), a dasyurid marsupial, should be one of them. Phascogales are easily managed in captivity, and there are now multiple studies involving their development, reproduction, nutrition, behavior and immune system, which can serve as a baseline for future studies. The addition of the phascogale as a model species will improve future mammalian studies by introducing variability and offer alternate solutions to biological problems, particularly in the areas of genetics, nutrition, immunology, the neuro-endocrine system, and ageing, due to their semelparous reproductive strategy and hence, subsequent predictive physiology. In this review, we provide information based on existing research on red-tailed phascogales to support their inclusion as a model species.

Clinical ophthalmological diagnostic description of 10 healthy sugar gliders (Petaurus breviceps) and prevalence of ocular-related presentations in a larger hospital population.

OBJECTIVE: To provide reference values for ocular examination and diagnostics in ophthalmologically normal sugar gliders (Petaurus breviceps). To retrospectively determine the prevalence of ocular diseases in sugar gliders presenting to a single institution. ANIMALS: Ten client owned and 106 previously evaluated sugar gliders. PROCEDURE: A descriptive study evaluated sugar gliders presented to Colorado State University's Avian, Exotics, and Zoological Medicine Service (CSU-AEZ) from August-2019 to January-2020. A complete ophthalmic examination including Schirmer tear test II (STT II), phenol red threat test (PRTT), intraocular pressure (IOP) via rebound tonometry, fluorescein, and rose bengal stain was performed under anesthesia. Conjunctival aerobic culture swabs and cytology were collected prior to ophthalmic evaluation. A retrospective review of medical records of sugar gliders presented to CSU-AEZ from 2008 to 2018 for ocular disease was performed. RESULTS: Mean values ± standard deviation for selected diagnostics included the following: STT II: 2.2 ± 6.7 mm/min; PRTT: 0 ± 0 mm/15 s; IOP: 12 ± 2.6 mm Hg. Fluorescein and rose bengal staining highlighted corneal abrasions secondary to tear testing. The three most common conjunctival bacterial isolates cultured were Staphylococcus spp. (3/20, 15%), Coryneform spp. (3/20, 15%), and unidentified Gram-positive cocci (3/20, 15%). Retrospective analysis revealed ocular diseases to be the third most common abnormality resulting in sugar glider presentations (13/106, 12.3%). CONCLUSION: This descriptive study gives reference values for IOP, conjunctival microbiology, and cytology for sugar gliders. STT II and PRTT provide little clinical value in sugar gliders. The retrospective study revealed that ocular abnormalities, often secondary to dental disease, are a common reason for presentation.

Finding food in a novel environment: The diet of a reintroduced endangered meso-predator to mainland Australia, with notes on foraging behaviour.

Translocated captive-bred predators are less skilled at hunting than wild-born predators and more prone to starvation post-release. Foraging in an unfamiliar environment presents many further risks to translocated animals. Knowledge of the diet and foraging behaviour of translocated animals is therefore an important consideration of reintroductions. We investigated the diet of the endangered meso-predator, the eastern quoll Dasyurus viverrinus. We also opportunistically observed foraging behaviour, enabling us to examine risks associated with foraging. Sixty captive-bred eastern quolls were reintroduced to an unfenced reserve on mainland Australia (where introduced predators are managed) over a two year period (2018, 2019). Quolls were supplementary fed macropod meat but were also able to forage freely. Dietary analysis of scats (n = 56) revealed that quolls ate macropods, small mammals, birds, invertebrates, fish, reptiles and frogs, with some between-year differences in the frequency of different diet categories. We also observed quolls hunting live prey. Quolls utilised supplementary feeding stations, indicating that this may be an important strategy during the establishment phase. Our study demonstrated that, in a novel environment, captive-bred quolls were able to locate food and hunt live prey. However, foraging was not without risks; with the ingestion of toxic substances and foraging in dangerous environments found to be potentially harmful. Knowledge of the diet of reintroduced fauna in natural landscapes is important for understanding foraging behaviour and evaluating habitat suitability for future translocations and management.

Infectious disease and sickness behaviour: tumour progression affects interaction patterns and social network structure in wild Tasmanian devils.

Infectious diseases, including transmissible cancers, can have a broad range of impacts on host behaviour, particularly in the latter stages of disease progression. However, the difficulty of early diagnoses makes the study of behavioural influences of disease in wild animals a challenging task. Tasmanian devils (Sarcophilus harrisii) are affected by a transmissible cancer, devil facial tumour disease (DFTD), in which tumours are externally visible as they progress. Using telemetry and mark-recapture datasets, we quantify the impacts of cancer progression on the behaviour of wild devils by assessing how interaction patterns within the social network of a population change with increasing tumour load. The progression of DFTD negatively influences devils' likelihood of interaction within their network. Infected devils were more active within their network late in the mating season, a pattern with repercussions for DFTD transmission. Our study provides a rare opportunity to quantify and understand the behavioural feedbacks of disease in wildlife and how they may affect transmission and population dynamics in general.