It takes two to tango: Phagocyte and lymphocyte numbers in a small mammalian hibernator.

Immunity is energetically costly and competes for resources with other physiological body functions, which may result in trade-offs that impair fitness during demanding situations. Endocrine mediators, particularly stress hormones, play a central role in these relationships and directly impact leukocyte differentials. To determine the effects of external stressors, energetic restraints and competing physiological functions on immune parameters and their relevance for fitness, we investigated leukocyte profiles during the active season of a small obligate hibernator, the edible dormouse (Glis glis), in five different study sites in south-western Germany. The highly synchronized yearly cycle of this species and the close adaptation of its life history to the irregular abundance of food resources provide a natural experiment to elucidate mechanisms underlying variations in fitness parameters. In contrast to previous studies on hibernators, that showed an immediate recovery of all leukocyte subtypes upon emergence, our study revealed that hibernation results in depleted phagocyte (neutrophils and monocytes) stores that recovered only slowly. As the phenomenon of low phagocyte counts was even more pronounced at the beginning of a low food year and primarily immature neutrophils were present in the blood upon emergence, preparatory mechanisms seem to determine the regeneration of phagocytes before hibernation is terminated. Surprisingly, the recovery of phagocytes thereafter took several weeks, presumably due to energetic restrictions. This impaired first line of defense coincides with lowest survival probabilities during the annual cycle of our study species. Reduced survival could furthermore be linked to drastic increases in the P/L ratio (phagocytes/lymphocytes), an indicator of physiological stress, during reproduction. On the other hand, moderate augmentations in the P/L ratio occurred during periods of low food availability and were associated with increased survival, but reproductive failure. In this case, the stress response probably represents an adaptive reaction that contributes to survival by activating energy resources. In contrast to our expectation, we could not detect an amplification of stress through high population densities. Summarized, results of our study clearly reveal that the leukocyte picture of active edible dormice responds sensitively to physiological conditions associated with hibernation, reproductive activity and food availability and can be linked to fitness parameters such as survival. Thus edible dormice represent an excellent model organism to investigate regulatory mechanisms of the immune system under natural conditions.

The vector tick Ixodes ricinus feeding on an arboreal rodent-the edible dormouse Glis glis.

The reservoir competence and long life expectancy of edible dormice, Glis glis, suggest that they serve as efficient reservoir hosts for Lyme disease (LD) spirochetes. Their arboreality, however, may reduce the probability to encounter sufficient questing Ixodes ricinus ticks to acquire and perpetuate LD spirochetes. To define the potential role of this small arboreal hibernator in the transmission cycle of LD spirochetes, we examined their rate and density of infestation with subadult ticks throughout the season of activity. Of the 1081 edible dormice that we captured at five study sites in Southern Germany and inspected for ticks at 2946 capture occasions, 26 % were infested with at least one and as many as 26 subadult ticks on their ear pinnae. The distribution of ticks feeding on edible dormice was highly aggregated. Although only few individuals harbored nymphal ticks soon after their emergence from hibernation, the rate of nymphal infestation increased steadily throughout the season and reached about 35 % in September. Dormice inhabiting a site with few conspecifics seemed more likely to be infested by numerous ticks, particularly nymphs, than those individuals living in densely populated sites. Male dormice were more likely to be parasitized by numerous nymphs than were females, independent of their age and body mass. Our observation that season, population density, and sex affect the rates of ticks feeding on edible dormice suggests that the contribution of edible dormice to the transmission cycle of LD spirochetes depends mainly on their ranging behavior and level of activity.

Ways to measure body temperature in the field.

Body temperature (Tb) represents one of the key parameters in ecophysiological studies with focus on energy saving strategies. In this study we therefore comparatively evaluated the usefulness of two types of temperature-sensitive passive transponders (LifeChips and IPTT-300) and one data logger (iButton, DS1922L) mounted onto a collar to measure Tb in the field. First we tested the accuracy of all three devices in a water bath with water temperature ranging from 0 to 40°C. Second, we evaluated the usefulness of the LifeChips and the modified iButtons for measuring Tb of small heterothermic mammals under field conditions. For this work we subcutaneously implanted 14 male edible dormice (Glis glis) with transponders, and equipped another 14 males with data loggers to simultaneously record Tb and oxygen consumption with a portable oxygen analyzer (Oxbox). In one individual we recorded Tb with both devices and analyzed recorded Tb patterns. LifeChips are able to measure temperature within the smallest range from 25 to 40°C with an accuracy of 0.07±0.12°C. IPTT-300 transponders measured temperature between 10 and 40°C, but accuracy decreased considerably at values below 30°C, with maximal deviations of nearly 7°C. An individual calibration of each transponder is therefore needed, before using it at low Tbs. The accuracy of the data logger was comparatively good (0.12±0.25°C) and stable over the whole temperature range tested (0-40°C). In all three devices, the repeatability of measurements was high. LifeChip transponders as well as modified iButtons measured Tb reliably under field conditions. Simultaneous Tb-recordings in one edible dormouse with an implanted LifeChip and a collar-mounted iButton revealed that values of both measurements were closely correlated. Taken together, we conclude that implanted temperature-sensitive transponders represent an appropriate and largely non-invasive method to measure Tb also under field conditions.

Functional morphology and bite performance of raptorial chelicerae of camel spiders (Solifugae).

Solifugae are an understudied group of relatively large arachnids with well over 1000 species distributed on almost all major continents. These highly active predators utilize their large chelicerae for feeding, defense, burrowing and mating. We investigated the differences in cheliceral morphology and performance of two ecologically divergent species from North Africa; the cursorial Galeodes sp. and the burrowing Rhagodes melanus. Morphological data show differences in aspect ratio between the two species. Bite force measurements show Rhagodes (N=11) to be a much stronger biter than Galeodes (N=8), in terms of both absolute maximum force (Rhagodes 5.63 N, Galeodes 2.12 N) and force relative to cheliceral size. Synchrotron microtomographs of one specimen for each species reveal large differences in physiological cross-sectional area (PCSA) and estimated muscle stress, resulting in a much higher muscle stress in Rhagodes. This species also showed a longer muscle fiber length. Muscle volume and PCSA were found to differ between the two chelicerae in the two scanned specimens. Whereas Rhagodes reflects this morphological asymmetry in having a higher bite force in the right chelicera, Galeodes shows no such bias.

Bridging environment, physiology and life history: Stress hormones in a small hibernator.

Our knowledge of the perception of stress and its implications for animals in the wild is limited, especially in regard to mammals. The aim of this study was therefore to identify sex specific effects of reproductive activity, body mass, food availability and hibernation on stress hormone levels in the edible dormouse (Glis glis), a small mammalian hibernator. Results of our study reveal that reproductive activity and pre-hibernation fattening were associated with high cortisol levels in both sexes. During the mating season, in particular individuals with low body masses had higher stress levels. Elevated levels of cortisol during pre-hibernation fattening were even higher in females that had formerly invested into reproduction compared to non-reproductive females. Previously observed impairments on health parameters and reduced survival rates associated with reproduction emphasize the functional relevance of high stress hormone levels for fitness. Prolonged food limitation, however, did not affect stress levels demonstrating the ability of dormice to predict and cope with food restriction.

Life history written in blood: erythrocyte parameters in a small hibernator, the edible dormouse.

The oxygen delivery system is one major determinant of the performance of vertebrates and responds sensitively to a variety of internal and environmental factors. To understand physiological mechanisms underlying variations of fitness, we investigated effects of demanding conditions associated with certain life-history events, food availability, and population density on the oxygen delivery system in free-ranging edible dormice (Glis glis). We sampled blood (n = 248) and urine (n = 319), performed an erythrocyte haemogram and visually determined the presence of haemoglobinuria. Reproduction leads to increased mortality in edible dormice and our study now reveals severe haematological impairments during reproduction that were associated with nutrient and energy deficits and stress. These effects were even more pronounced in subsequent reproductive years, indicating prolonged physiological impairment. Under limited food availability, the rate of erythrocyte generation was reduced. This seems to be part of an energy saving strategy instead of representing a poor body condition as survival probability in this species is high in years of low food availability. A high prevalence ratio of haemoglobinuria (up to 85%) at the end of the active season indicated amplified erythrocyte destruction through haemolysis. This may be the result of a preparative mechanism to avoid massive oxidative damage during the long hibernation period. Most ecophysiological studies so far focus on single erythrocyte parameters on a short time scale, which could be misleading. Our results clearly highlight that a wide-array RBC approach is a powerful tool for investigating mechanisms underlying physiological performance and fitness, also for other vertebrate taxa.

Flexibility is the key: metabolic and thermoregulatory behaviour in a small endotherm.

Hibernation is the most effective way to reduce thermoregulatory costs during periods of unfavourable environmental conditions. In preparation to hibernation, fat-storing hibernators accumulate large quantities of body fat, which increases their locomotor costs and also the risk of predation. As a consequence, there should be a strong selective pressure to restrict pre-hibernation fattening to a short-time period before the onset of hibernation. The edible dormouse (Glis glis) is characterized by having adapted its whole life history to the irregularly occurring mast-seeding pattern of the European beech (Fagus sylvaticus). Thus, the question arises how this small endotherm copes with huge differences in food availability between years. Therefore, we investigated body mass and thermal energetics of edible dormice during high and low food years. Our results demonstrate that during periods of low food availability, edible dormice enter an energy-saving mode with reduced body temperature (Tb) and resting metabolic rate (RMR), and high torpor frequencies. During irregularly occurring short events of high food availability in mast years, however, Tb was higher, torpor did not occur, and RMR was drastically elevated possibly due to an enlarged digestive tract and the heat increment of feeding associated with a dietary switch to high-quality food and an increase in the amount of food ingested. This physiological flexibility allows edible dormice to efficiently accumulate body fat reserves under extremely different situations of food availability.