Late Hibernation Research Articles

Disparities in Perimyotis subflavus Body Mass Between Cave and Culvert Hibernacula in Georgia, USA.

The tricolored bat (Perimyotis subflavus), once common in the eastern United States, has experienced significant mortality due to white-nose syndrome (WNS), a fungal disease that primarily affects bats hibernating in caves and mines. In coastal regions of the southeastern United States, where caves and mines are scarce, tricolored bats often use roadway culverts as hibernacula. However, WNS infection dynamics in culverts are poorly understood. Previous research indicated that bats with higher body mass at the onset of hibernation have a higher probability of surviving repeated arousal events from WNS. Therefore, we compared tricolored bat winter body mass between cave and culvert hibernacula and identified culvert characteristics influencing body mass during hibernation in Georgia, USA. From 2018 to 2022, we measured body mass of 754 individuals in early and late hibernation across 32 culverts (n = 497) and four caves (n = 257). Our study revealed a southward spread of the fungus over multiple years, with the first confirmed case of WNS in a Georgia culvert in 2022. Overall, tricolored bats in caves weighed more in early hibernation than those in culverts, but bats in culverts weighed more in late hibernation. Across all sites, female tricolored bats entering and leaving hibernation had greater mass than males but lost more mass during hibernation, possibly due to differences in torpor-arousal patterns and WNS infection rates. Additionally, all bats lost more mass in longer culverts. Understanding culvert characteristics affecting bat body mass will inform management strategies to mitigate WNS effects. Identifying risk factors for specific tricolored bat hibernacula can guide managers on where to focus winter WNS monitoring efforts and potential treatments.

Microbiota diversity and anti-Pseudogymnoascus destructans bacteria isolated from Myotis pilosus skin during late hibernation.

Symbiotic microorganisms that reside on the host skin serve as the primary defense against pathogens in vertebrates. Specifically, the skin microbiome of bats may play a crucial role in providing resistance against Pseudogymnoascus destructans (Pd), the pathogen causing white-nose syndrome. However, the epidermis symbiotic microbiome and its specific role in resisting Pd in highly resistant bats in Asia are still not well understood. In this study, we collected and characterized skin microbiota samples of 19 Myotis pilosus in China and explored the differences between Pd-positive and negative individuals. We identified inhibitory effects of these bacteria through cultivation methods. Our results revealed that the Simpson diversity index of the skin microbiota for positive individuals was significantly lower than that of negative individuals, and the relative abundance of Pseudomonas was significantly higher in positive bats. Regardless of whether individuals were positive or negative for Pd, the relative abundance of potentially antifungal genera in skin microbiota was high. Moreover, we successfully isolated 165 microbes from bat skin and 41 isolates from positive individuals able to inhibit Pd growth compared to only 12 isolates from negative individuals. A total of 10 genera of Pd-inhibiting bacteria were screened, among which the genera Algoriella, Glutamicibacter, and Psychrobacter were newly discovered as Pd-inhibiting genera. These Pd-inhibiting bacteria metabolized a variety of volatile compounds, including dimethyl trisulfide, dimethyl disulfide, propylene sulfide, 2-undecanone, and 2-nonanone, which were able to completely inhibit Pd growth at low concentrations.IMPORTANCERecently, white-nose syndrome has caused the deaths of millions of hibernating bats, even threatening some with regional extinction. Bats in China with high resistance to Pseudogymnoascus destructans can provide a powerful reference for studying the management of white-nose syndrome and understanding the bats against the pathogen's intrinsic mechanisms. This study sheds light on the crucial role of host symbiotic skin microorganisms in resistance to pathogenic fungi and highlights the potential for harnessing natural defense mechanisms for the prevention and treatment of white-nose syndrome. In addition, this may also provide promising candidates for the development of bioinsecticides and fungicides that offer new avenues for addressing fungal diseases in wildlife and agricultural environments.

Non-shivering thermogenesis is differentially regulated during the hibernation season in Arctic ground squirrels.

Arctic ground squirrels are small mammals that experience physiological extremes during the hibernation season. Body temperature rises from 1°C to 40°C during interbout arousal and requires tight thermoregulation to maintain rheostasis. Tissues from wild-caught Arctic ground squirrels were sampled over 9months to assess the expression of proteins key to thermogenic regulation. Animals were sacrificed while aroused, and the extensor digitorum longus, diaphragm, brown adipose tissue, and white adipose tissue were probed using Western blots to assess protein expression and blood was sampled for metabolite analysis. Significant seasonal expression patterns emerged showing differential regulation. Contrary to our prediction, white adipose tissue showed no expression of uncoupling protein 1, but utilization of uncoupling protein 1 peaked in brown adipose tissue during the winter months and began to taper after terminal arousal in the spring. The opposite was true for muscular non-shivering thermogenesis. Sarco/endoplasmic reticulum calcium ATPase 1a and 2a expressions were depressed during the late hibernation season and rebounded after terminal arousal in diaphragm tissues, but only SERCA2a was differentially expressed in the extensor digitorum longus. The uncoupler, sarcolipin, was only detected in diaphragm samples and had a decreased expression during hibernation. The differential timing of these non-shivering pathways indicated distinct functions in maintaining thermogenesis which may depend on burrow temperature, availability of endogenous resources, and other seasonal activity demands on these tissues. These results could be impacted by fiber type makeup of the muscles collected, the body weight of the animal, and the date of entrance or exit from hibernation.

Effects of Spring Warming on Seasonal Neuroendocrinology and Activation of the Reproductive Axis in Hibernating Arctic Ground Squirrels.

Many animals adjust the timing of seasonal events, such as reproduction, molt, migration, and hibernation, in response to interannual variation and directional climate-driven changes in temperature. However, the mechanisms by which temperature influences seasonal timing are relatively under-explored. Seasonal timing involves retrograde signaling in which thyrotropin (TSH) in the pars tuberalis (PT) alters expression of thyroid hormone (TH) deiodinases (Dio2/Dio3) in tanycyte cells lining the third ventricle of the hypothalamus. This, in turn, affects the availability of triiodothyronine (T3) within the mediobasal hypothalamus-increased hypothalamic T3 restores a summer phenotype and activates the reproductive axis in long-day breeders. Recently, we showed that retrograde TH signaling is activated during late hibernation in arctic ground squirrels (Urocitellus parryii) held in constant darkness and constant ambient temperature. Sensitivity of seasonal pathways to nonphotic cues, such as temperature, is likely particularly important to hibernating species that are sequestered in hibernacula during spring. To address this issue, we exposed captive arctic ground squirrels of both sexes to an ecologically relevant increase in ambient temperature (from -6 to -1°C) late in hibernation and examined the effects of warming on the seasonal retrograde TSH/Dio/T3 signaling pathway, as well as downstream elements of the reproductive axis. We found that warmed males tended to have higher PT TSHβ expression and significantly heavier testis mass whereas the TSH/Dio/T3 signaling pathway was unaffected by warming in females, although warmed females exhibited a slight decrease in ovarian mass. Our findings suggest that temperature could have different effects on gonadal growth in male and female arctic ground squirrels, which could lead to mismatched timing in response to rapid climate change.

Emergence activity at hibernacula differs among four bat species affected by white-nose syndrome.

Prior to the introduction of white‐nose syndrome (WNS) to North America, temperate bats were thought to remain within hibernacula throughout most of the winter. However, recent research has shown that bats in the southeastern United States emerge regularly from hibernation and are active on the landscape, regardless of their WNS status. The relationship between winter activity and susceptibility to WNS has yet to be explored but warrants attention, as it may enable managers to implement targeted management for WNS‐affected species. We investigated this relationship by implanting 1346 passive integrated transponder (PIT) tags in four species that vary in their susceptibility to WNS. Based on PIT‐tag detections, three species entered hibernation from late October to early November. Bats were active at hibernacula entrances on days when midpoint temperatures ranged from −1.94 to 22.78°C (mean midpoint temperature = 8.70 ± 0.33°C). Eastern small‐footed bats (Myotis leibii), a species with low susceptibility to WNS, were active throughout winter, with a significant decrease in activity in mid‐hibernation (December 16 to February 15). Tricolored bats (Perimyotis subflavus), a species that is highly susceptible to WNS, exhibited an increase in activity beginning in mid‐hibernation and extending through late hibernation (February 16 to March 31). Indiana bats (M. sodalis), a species determined to have a medium–high susceptibility to WNS, remained on the landscape into early hibernation (November 1 to December 15), after which we did not record any again until the latter portion of mid‐hibernation. Finally, gray bats (M. grisescens), another species with low susceptibility to WNS, maintained low but regular levels of activity throughout winter. Given these results, we determined that emergence activity from hibernacula during winter is highly variable among bat species and our data will assist wildlife managers to make informed decisions regarding the timing of implementation of species‐specific conservation actions.

Phenotypic changes in the metabolic profile and adiponectin activity during seasonal fattening and hibernation in female Daurian ground squirrels (Spermophilus dauricus).

Seasonal hibernation has provided an opportunity to study animals' phenotypic plasticity in adaptation to changing environment. In the present study focusing on the female Daurian ground squirrel (Spermophilus dauricus)-a well demonstrated seasonal hibernator-we examined their behavioral, morphological, and metabolic changes during fattening, hibernation, and emergence. Our data indicated high levels of food intake, fat deposition, and body mass increases during fattening compared to hibernation. The levels of serum glucose and triglycerides were also higher during fattening than during hibernation and emergence. Interestingly, although squirrels showed signs of obesity and elevated triglycerides in serum during fattening, triglyceride levels in the liver and skeletal muscles remained unchanged. Our data also indicated that adiponectin levels in serum and cerebrospinal fluid were different between fattening and hibernation. Levels of adiponectin receptor 1 in the skeletal muscle remained low during fattening but peaked in late hibernation. In contrast, adiponectin receptor 2 in the liver showed a steady increase during fattening, which was followed by a significant decrease at early hibernation. Our data indicate that adiponectin may play an important role in preventing heterotopic fat accumulation in a receptor- and organ-specific manner, as well as in facilitating the switch from glucose metabolism to lipid metabolism during fattening and hibernation in female Daurian ground squirrels.

MicroRNA dynamics during hibernation of the Australian central bearded dragon (Pogona vitticeps)

Hibernation is a physiological state employed by many animals that are exposed to limited food and adverse winter conditions. Controlling tissue-specific and organism wide changes in metabolism and cellular function requires precise regulation of gene expression, including by microRNAs (miRNAs). Here we profile miRNA expression in the central bearded dragon (Pogona vitticeps) using small RNA sequencing of brain, heart, and skeletal muscle from individuals in late hibernation and four days post-arousal. A total of 1295 miRNAs were identified in the central bearded dragon genome; 664 of which were novel to central bearded dragon. We identified differentially expressed miRNAs (DEmiRs) in all tissues and correlated mRNA expression with known and predicted target mRNAs. Functional analysis of DEmiR targets revealed an enrichment of differentially expressed mRNA targets involved in metabolic processes. However, we failed to reveal biologically relevant tissue-specific processes subjected to miRNA-mediated regulation in heart and skeletal muscle. In brain, neuroprotective pathways were identified as potential targets regulated by miRNAs. Our data suggests that miRNAs are necessary for modulating the shift in cellular metabolism during hibernation and regulating neuroprotection in the brain. This study is the first of its kind in a hibernating reptile and provides key insight into this ephemeral phenotype.

Torpor expression is associated with differential spermatogenesis in hibernating eastern chipmunks.

Hibernators suppress physiological processes when expressing torpor, yet little is known about the effects of torpor on male reproductive physiology. Studies of hibernating mammals suggest that deep torpor negatively impacts spermatogenesis and that transitions between torpor and euthermic arousals increase cellular oxidative stress, with potentially damaging effects on sperm. Here, we hypothesize that variation in torpor expression affects the reproductive readiness of hibernators by impacting their sperm production. To test this, we examined the relationship between torpor expression and spermatogenesis in captive eastern chipmunks (Tamias striatus). We determined torpor depth with temperature data loggers and assessed its relationship with spermatogenesis by examining spermatogenic progression, cell division, sperm counts, sperm maturity, and DNA damage. We show that deep hibernators (high levels of torpor) largely halted spermatogenesis in late hibernation in comparison with shallow hibernators (low levels of torpor), where ongoing spermatogenesis was observed. Despite these differences in spermatogenic state during hibernation, spermatogenic progression, sperm numbers, and maturity did not differ in spring, potentially reflecting similar degrees of reproductive readiness. Interestingly, shallow hibernators exhibited higher rates of DNA damage in spermatogenic cells during hibernation, with this trend reversing in spring. Our results thus indicate that once heterothermy is terminated, deep hibernators resume spermatogenesis but are characterized by higher rates of DNA damage in spermatogenic cells at the seasonal stage when spring mating commences. Therefore, our study confirmed posthibernation recovery of sperm production but also a potential impact of deep torpor expression during winter on DNA damage in spring.

Cardiac adaptation in hibernating, free-ranging Scandinavian Brown Bears (Ursus arctos)

During six months of annual hibernation, the brown bear undergoes unique physiological changes to adapt to decreased metabolic rate. We compared cardiac structural and functional measures of hibernating and active bears using comprehensive echocardiography. We performed echocardiography on 13 subadult free-ranging, anaesthetised Scandinavian brown bears (Ursus arctos) during late hibernation and in early summer. Mean heart rate was 26 beats per minute (standard deviation (SD): 8) during hibernation vs 71 (SD: 14) during active state. All left ventricular (LV) systolic and diastolic measures were decreased during hibernation: mean ejection fraction: 44.2% (SD: 6.0) active state vs 34.0 (SD: 8.1) hibernation, P = 0.001; global longitudinal strain: −11.2% (SD: 2.0) vs −8.8 (SD: 3.3), P = 0.03; global longitudinal strain rate: −0.82 (SD: 0.15) vs −0.41 (SD: 0.18), P < 0.001; septal e’: 9.8 cm/s (SD: 1.8) vs 5.2 (SD: 2.7), P < 0.001. In general, measures of total myocardial motion (ejection fraction and global longitudinal strain) were decreased to a lesser extent than measures of myocardial velocities. In the hibernating brown bear, cardiac adaptation included decreased functional measures, primarily measures of myocardial velocities, but was not associated with cardiac atrophy. Understanding the mechanisms of these adaptations could provide pathophysiological insight of human pathological conditions such as heart failure.

Waking the sleeping dragon: gene expression profiling reveals adaptive strategies of the hibernating reptile Pogona vitticeps

BackgroundHibernation is a physiological state exploited by many animals exposed to prolonged adverse environmental conditions associated with winter. Large changes in metabolism and cellular function occur, with many stress response pathways modulated to tolerate physiological challenges that might otherwise be lethal. Many studies have sought to elucidate the molecular mechanisms of mammalian hibernation, but detailed analyses are lacking in reptiles. Here we examine gene expression in the Australian central bearded dragon (Pogona vitticeps) using mRNA-seq and label-free quantitative mass spectrometry in matched brain, heart and skeletal muscle samples from animals at late hibernation, 2 days post-arousal and 2 months post-arousal.ResultsWe identified differentially expressed genes in all tissues between hibernation and post-arousal time points; with 4264 differentially expressed genes in brain, 5340 differentially expressed genes in heart, and 5587 differentially expressed genes in skeletal muscle. Furthermore, we identified 2482 differentially expressed genes across all tissues. Proteomic analysis identified 743 proteins (58 differentially expressed) in brain, 535 (57 differentially expressed) in heart, and 337 (36 differentially expressed) in skeletal muscle. Tissue-specific analyses revealed enrichment of protective mechanisms in all tissues, including neuroprotective pathways in brain, cardiac hypertrophic processes in heart, and atrophy protective pathways in skeletal muscle. In all tissues stress response pathways were induced during hibernation, as well as evidence for gene expression regulation at transcription, translation and post-translation.ConclusionsThese results reveal critical stress response pathways and protective mechanisms that allow for maintenance of both tissue-specific function, and survival during hibernation in the central bearded dragon. Furthermore, we provide evidence for multiple levels of gene expression regulation during hibernation, particularly enrichment of miRNA-mediated translational repression machinery; a process that would allow for rapid and energy efficient reactivation of translation from mature mRNA molecules at arousal. This study is the first molecular investigation of its kind in a hibernating reptile, and identifies strategies not yet observed in other hibernators to cope stress associated with this remarkable state of metabolic depression.

Numerous cold arousals and rare arousal cascades as a hibernation strategy in European Myotis bats.

Hibernating bats optimise the duration of torpor bouts and arousals in relation to hibernaculum microclimatic conditions and fat reserves. Clustering has significant physiological and ecological benefits, promoting successful hibernation of individuals. Such aggregations may help maintain optimal temperatures, allowing better energy utilisation than in solitarily bats. However, aroused bats in a cluster could conceivably disturb those still hibernating, starting an energy-demanding arousal process. Our study was conducted over two winters in two different hibernacula (cave and mine) in the Czech Republic, where Greater mouse-eared bats (Myotis myotis) have previously been diagnosed with white-nose syndrome. In 118 arousal episodes we recorded 193 individual arousals in which a warming phase was observed, 135 (69.9%) being cold arousals, where bats ceased increasing their body temperatures at ≤ 10 °C. The remaining arousals were standard normothermic arousals, where body (fur) surface temperatures reached > 20 °C. Cold arousals occurred during the mid- and late hibernation periods, suggesting they were a response to disturbance by a neighbour in the same cluster. Arousal cascades, where bats aroused in series, were rare (12.7%) and reached a maximum in mid-January. Our data suggest that Myotis bats prolong their torpor bouts using numerous cold arousals but few arousal cascades. Upon arrival of a bat, the clustered bats show tolerance to disturbing by conspecifics.

Seasonal Shifts in Gut Microbiota‐Mediated Nitrogen Recycling in 13‐Lined Ground Squirrels

Hibernation is a complex phenotype that enables animals to fast for several months during periods of food scarcity. During this time, less of the hibernator's protein is degraded than would be predicted and, in the 13‐lined ground squirrel (Ictidomys tridecemlineatus), muscle volume and protein synthesis rise near the end of the hibernation season despite continued fasting and loss of body mass. With no exogenous nitrogen intake, one nitrogen source to support protein synthesis in late winter could be from urea nitrogen salvage (UNS), a nitrogen recycling process mediated by the ureolytic activity of the hibernator's gut microbiota. We hypothesized that the contribution of UNS to skeletal muscle growth in 13‐lined ground squirrels increases over the hibernation season, and tested this by comparing UNS processes in summer, early hibernation (1 month), and late hibernation (&gt;3 months) squirrels. Data collected thus far suggest that expression of the urea transporter (UT‐B) in cecal tissue is 3‐fold greater in late winter than in summer squirrels (P&lt;0.001; n=12 per group), indicating a greater capacity for transport of urea into the cecum during winter fasting. Real‐time measurement of microbial ureolytic activity using cavity ring‐down spectrometry on squirrels I.P.‐injected with 13C,15N‐labeled urea confirmed the bacterial origin of UNS, because δ13C in breath is substantially reduced in antibiotic‐depleted microbiotas. Isotope ratio mass spectrometry showed that tissues of squirrels treated with 13C,15N‐urea accumulated up to 200x more 15N as saline‐treated control squirrels, confirming that urea‐derived nitrogen is recycled and incorporated into host tissues. Together, these results corroborate UNS in active and hibernating 13‐lined ground squirrels and suggest that this mechanism for nitrogen recycling is a key aspect of the host‐microbe symbiosis in hibernating mammals that helps prepare the squirrels for spring emergence.Support or Funding InformationThis work is supported by NSF award IOS1558044This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Tissue-specific telomere dynamics in hibernating arctic ground squirrels (Urocitellus parryii).

Hibernation is used by a variety of mammals to survive seasonal periods of resource scarcity. Reactive oxygen species (ROS) released during periodic rewarming throughout hibernation, however, may induce oxidative damage in some tissues. Telomeres, which are the terminal sequences of linear chromosomes, may shorten in the presence of ROS, and thus the telomere length of an individual reflects the degree of accrued oxidative damage. This study quantified telomere length dynamics throughout hibernation in arctic ground squirrels (Urocitellus parryii). We hypothesized that telomere dynamics are tissue specific and predicted that telomere shortening would be most pronounced in brown adipose tissue (BAT), the organ that directly supports non-shivering thermogenesis during arousals. We used qPCR to determine relative telomere length (RTL) in DNA extracted from liver, heart, skeletal muscle (SM) and BAT of 45 juvenile and adult animals sampled either at mid- or late hibernation. Age did not have a significant effect on RTL in any tissue. At mid-hibernation, RTL of juvenile females was longer in BAT and SM than in liver and heart. In juvenile females, RTL in BAT and SM, but not in liver and heart, was shorter at late hibernation than at mid-hibernation. At late hibernation, juvenile males had longer RTL in BAT than did juvenile females, perhaps due to the naturally shorter hibernation duration of male arctic ground squirrels. Finally, BAT RTL at late hibernation negatively correlated with arousal frequency. Overall, our results suggest that, in a hibernating mammal, telomere shortening is tissue specific and that metabolically active tissues might incur higher levels of molecular damage.

The other white-nose syndrome transcriptome: Tolerant and susceptible hosts respond differently to the pathogen Pseudogymnoascus destructans.

Mitigation of emerging infectious diseases that threaten global biodiversity requires an understanding of critical host and pathogen responses to infection. For multihost pathogens where pathogen virulence or host susceptibility is variable, host–pathogen interactions in tolerant species may identify potential avenues for adaptive evolution in recently exposed, susceptible hosts. For example, the fungus Pseudogymnoascus destructans causes white‐nose syndrome (WNS) in hibernating bats and is responsible for catastrophic declines in some species in North America, where it was recently introduced. Bats in Europe and Asia, where the pathogen is endemic, are only mildly affected. Different environmental conditions among Nearctic and Palearctic hibernacula have been proposed as an explanation for variable disease outcomes, but this hypothesis has not been experimentally tested. We report the first controlled, experimental investigation of response to P. destructans in a tolerant, European species of bat (the greater mouse‐eared bat, Myotis myotis). We compared body condition, disease outcomes and gene expression in control (sham‐exposed) and exposed M. myotis that hibernated under controlled environmental conditions following treatment. Tolerant M. myotis experienced extremely limited fungal growth and did not exhibit symptoms of WNS. However, we detected no differential expression of genes associated with immune response in exposed bats, indicating that immune response does not drive tolerance of P. destructans in late hibernation. Variable responses to P. destructans among bat species cannot be attributed solely to environmental or ecological factors. Instead, our results implicate coevolution with the pathogen, and highlight the dynamic nature of the “white‐nose syndrome transcriptome.” Interspecific variation in response to exposure by the host (and possibly pathogen) emphasizes the importance of context in studies of the bat‐WNS system, and robust characterization of genetic responses to exposure in various hosts and the pathogen should precede any attempts to use particular bat species as generalizable “model hosts.”

Oxidation of linoleic and palmitic acid in pre-hibernating and hibernating common noctule bats revealed by 13C breath testing.

Mammals fuel hibernation by oxidizing saturated and unsaturated fatty acids from triacylglycerols in adipocytes, yet the relative importance of these two categories as an oxidative fuel may change during hibernation. We studied the selective use of fatty acids as an oxidative fuel in noctule bats (Nyctalus noctula). Pre-hibernating noctule bats that were fed 13C-enriched linoleic acid (LA) showed 12 times higher tracer oxidation rates compared with conspecifics fed 13C-enriched palmitic acid (PA). After this experiment, we supplemented the diet of bats with the same fatty acids on five subsequent days to enrich their fat depots with the respective tracer. We then compared the excess 13C enrichment (excess atom percentage, APE) in breath of bats for torpor and arousal events during early and late hibernation. We observed higher APE values in breath of bats fed 13C-enriched LA than in bats fed 13C-enriched PA for both states (torpor and arousal), and also for both periods. Thus, hibernating bats selectively oxidized endogenous LA instead of PA, probably because of faster transportation rates of polyunsaturated fatty acids compared with saturated fatty acids. We did not observe changes in APE values in the breath of torpid animals between early and late hibernation. Skin temperature of torpid animals increased by 0.7°C between early and late hibernation in bats fed PA, whereas it decreased by -0.8°C in bats fed LA, highlighting that endogenous LA may fulfil two functions when available in excess: serving as an oxidative fuel and supporting cell membrane functionality.

Serum Biochemical Reference Values for Adult and Non-adult Chinese Alligators during the Deep and Late Hibernation Periods

Background: The Chinese alligator (Alligator sinensis) is a critically endangered species. Due to the rapid growth of the captive population, the susceptibility to disease during the recovery period after winter hibernation, especially in young alligators, have detrimentally affected Chinese alligator populations. Serum biochemistry, which relates to metabolism, nutritional status and disease, is enormously helpful in evaluating physical conditions in reptile. Many studies have reported the serum biochemical reference values of various reptilian species, including several crocodilians. However, reference values for Chinese alligators have not yet been reported. For captive Chinese alligators, hibernation is a crucial period because winter management has a direct influence on the survival rate of juveniles and the reproduction rate of adults. The main object of the present study refore was to measure the serum biochemical values of captive Chinese alligators during hibernation.Materials, Methods &amp; Results: As such, this study investigates the serum biochemistry as a factor of age and hibernation stage. During the deep and late hibernation periods blood samples were drawn from 30 healthy captive Chinese alligators (adults, sub-adults, and juveniles) at the Anhui Research Center of Chinese Alligator Reproduction (ARCCAR). Serum biochemical measurements were performed using an automated biochemical analyzer and compared based on the age group and hibernation stage via two-way ANOVA. During late hibernation, serum lactate dehydrogenase, alkaline phosphatase, and aspartate aminotransferase activity increased in all age groups in comparison to that in deep hibernation, while the concentration of calcium decreased. Meanwhile, the concentration of serum phosphorus, uric acid, total protein, and globulin in sub-adults and juveniles considerably increased in comparison to that in deep hibernation, while cholesterol and albumin declined. However, in adults only slight changes were noted. Based on comprehensive statistical analysis, our results indicate that sub-adults and juveniles are at risk of developing renal disease during artificial hibernation.Discussion: Chinese alligators, especially sub-adults and juveniles, are particularly vulnerable to disease when they wake from hibernation. They often display symptoms such as depression, anorexia, lethargy, sluggish movement, slow, incremental weight gain, progressive muscle wasting, and even death. The high rate of morbidity in non-adult Chinese alligators may be associated with the high density of UA and other changes in multiple biochemical markers that occur during late hibernation. These altered serum biochemical profiles may indicate kidney damage. One of the most common diseases among reptiles is nephropathy, the symptoms of which are non-specific and tend to agree with those observed post-hibernation. In summary, this study has reported the serum biochemical values of Chinese alligators of varying ages in the deep and late hibernation phases. Based on statistical analyses, interesting differences between the serum biochemical values of adults and non-adults during the deep and late hibernation have been found. The observed changes suggest that, under an artificial hibernation environment, the kidneys of sub-adults and juveniles may become impaired. We believe that the data reported in this study will provide clinical guidance to facilitate more appropriate artificial wintering conditions for Chinese alligators, and assist the breeding and management of these reptiles, as well as disease prevention, during hibernation and recovery.

How to spend the summer? Free-living dormice (Glis glis) can hibernate for 11 months in non-reproductive years.

Edible dormice are arboreal rodents adapted to yearly fluctuations in seed production of European beech, a major food source for this species. In years of low beech seed abundance, dormice skip reproduction and non-reproductive dormice fed ad libitum in captivity can display summer dormancy in addition to winter hibernation. To test whether summer dormancy, that is, a very early onset of hibernation, actually occurs in free-living dormice, we monitored core body temperature (Tb) over ~12 months in 17 animals during a year of beech seeding failure in the Vienna Woods. We found that 8 out of 17 dormice indeed re-entered hibernation as early as in June/July, with five of them having extreme hibernation durations of 11 months or more (total range: 7.8–11.4 months). Thus, we show for the first time that a free-living mammal relying on natural food resources can continuously hibernate for >11 months. Early onset of hibernation was associated with high body mass in the spring, but the distribution of hibernation onset was bimodal with prolonged hibernation starting either early (prior to July 28) or late (after August 30). This could not be explained by differences in body mass alone. Animals with a late hibernation onset continued to maintain high nocturnal Tb’s throughout summer but used short, shallow torpor bouts (mean duration 7.44 ± 0.9 h), as well as occasional multiday torpor for up to 161 h.Electronic supplementary materialThe online version of this article (doi:10.1007/s00360-015-0929-1) contains supplementary material, which is available to authorized users.

English

Since its 2009 detection in Virginia hibernacula, the fungal pathogen Pseudogymnoascus destructans causing White-nose Syndrome (WNS) has had a marked impact on cave bats locally. From 2008-2013, we documented numeric and physiologic changes in cave bats through fall swarm (FS), early hibernation (EH), and late hibernation (LH) capture and banding surveys at 18 hibernacula in western Virginia. We coupled active surveys with passive biennial winter counts in 2009, 2011, and 2013. We compared individual body mass index (BMI) across years for FS, EH, and LH hibernation to determine if WNS impacts on extant bats would be manifested by changes in body condition (as anecdotally observed elsewhere for WNS-impacted bats) as well as a population reduction. To estimate percent declines in bat presence or relative activity, we used FS capture per-unit-effort data, and the winter hibernacula absolute counts. We captured 4,524 bats of eight species, with species-specific capture success declining by 75-100% post-WNS. Little brown bats (Myotis lucifugus) exhibited the greatest declines in winter hibernacula counts (AVG. = 99.0% decline), followed by tri-colored bats (Perimyotis subflavus; 89.5% decline) and Indiana bats (M. sodalis; 33.5% decline). Graphical analyses of captures-per-trap-hour in FS showed declines for little brown bats, tri-colored bats, and northern long-eared bats (M. septentrionalis), but suggest a modest rebound of Indiana bat numbers. Fall swarm trends in BMI suggested some drops post-WNS exposure, but these trends were not consistent across sexes or seasonal time blocks. Our inconclusive BMI metrics and little brown bat band recapture data suggest little competitive advantage or selection for surviving bats. Lesser (but apparent) declines in Indiana bat numbers mirrors trends seen elsewhere regionally, and band recoveries do show that some individuals are persisting. Additional surveys will determine if bats in Virginia will persist or face extirpation due to presumed low recruitment and survivorship. Key words: Bat, hibernaculum, Pseudogymnoascus destructans, Virginia, white-nose syndrome.

Hematology of three age groups of captive Chinese alligator during deep and late hibernation

As the numbers and density of the farmed Chinese alligator (Alligator sinensis) increase, diseases are more common. Death often occurs during the recovery period from hibernation and this phenomenon is particularly prominent in juveniles. A practical way is therefore needed to monitor the health of the alligator. Research on hematology can provide an early warning and a rapid diagnosis of a disease and will help decrease mortality during recovery from hibernation. The aim of this study was to determine the hematologic values of the Chinese alligator at different stages of hibernation. We collected 30 blood samples from each of three age groups (adults, sub-adults, juveniles) during deep and late hibernation. We obtained counts of red blood cells, thrombocytes, and total and different white blood cells using manual methods. The thrombocyte numbers in the three groups decreased significantly during late compared with deep hibernation. The white blood cell counts for the sub-adults and juveniles increased significantly during late hibernation, whereas this did not occur in the adults. The immunity of the sub-adults and juveniles clearly increased during late hibernation. The results suggest that, for breeding programs, environmental stress may be the cause of the higher mortality in the juveniles during recovery from hibernation.

Identification and coordinated expression of perilipin genes in the biological cycle of sunn pest, Eurygaster maura (Hemiptera: Scutelleridae): Implications for lipolysis and lipogenesis

The sunn pest, Eurygaster spp., is one of the most destructive pests of grains in Asia, Europe and Africa. The nymphs and adults feed voraciously in the field by late-spring, followed by migration of adults into mountains for diapause, which includes estivation by late summer and hibernation during winter. Adults migrate back to the field by the end of diapause in mid-spring, where they mate and lay eggs. To understand how sunn pest survives and maintains basic metabolic functions without feeding for 7 months during diapause, this study focused on lipid metabolism as the major source of energy production, and the primary organ of lipid metabolism, the fat body. Studies on lipid metabolism revealed two major factors referred to perilipin protein family, Lipid Storage Droplet Protein 1 (LSD1) and Lipid Storage Droplet Protein 2 (LSD2), which are involved in hydrolysis and accumulation of lipids, respectively. In this study, two LSD (EmLSD1-2) orthologues in the hemimetabolous Eurygaster maura were identified. EmLSD1 and EmLSD2 genes were expressed in multiple tissues, but primarily in fat body. Both genes were continuously expressed throughout the insect's life cycle but peaked in the 4th nymphal stage. Their expression patterns were in accordance with the biological roles of LSDs. EmLSD1 expression peaked in non-feeding stages supporting its lipolytic role, while the highest level of EmLSD2 expression was in feeding stages supporting its lipogenetic role. Expression patterns of both genes differed in females and males. Overall, expression patterns of EmLSDs provide clues to understanding the interesting life cycle of sunn pest.