Disease recovery in bats affected by white-nose syndrome.

Processes associated with recovery of survivors are understudied components of wildlife infectious diseases. White-nose syndrome (WNS) in bats provides an opportunity to study recovery of disease survivors, understand implications of recovery for individual energetics, and assess the role of survivors in pathogen transmission. We documented temporal patterns of recovery from WNS in little brown bats (Myotis lucifugus) following hibernation to test the hypotheses that: (1) recovery of wing structure from WNS matches a rapid time scale (i.e. approximately 30 days) suggested by data from free-ranging bats; (2) torpor expression plays a role in recovery; (3) wing physiological function returns to normal alongside structural recovery; and (4) pathogen loads decline quickly during recovery. We collected naturally infected bats at the end of hibernation, brought them into captivity, and quantified recovery over 40 days by monitoring body mass, wing damage, thermoregulation, histopathology of wing biopsies, skin surface lipids and fungal load. Most metrics returned to normal within 30 days, although wing damage was still detectable at the end of the study. Torpor expression declined overall throughout the study, but bats expressed relatively shallow torpor bouts - with a plateau in minimum skin temperature - during intensive healing between approximately days 8 and 15. Pathogen loads were nearly undetectable after the first week of the study, but some bats were still detectably infected at day 40. Our results suggest that healing bats face a severe energetic imbalance during early recovery from direct costs of healing and reduced foraging efficiency. Management of WNS should not rely solely on actions during winter, but should also aim to support energy balance of recovering bats during spring and summer.

Sonar strobe groups and buzzes are produced before powered flight is achieved in the juvenile big brown bat, Eptesicus fuscus.

Laryngeally echolocating bats produce a rapid succession of echolocation calls just before landing. These landing buzzes exhibit an increase in call rate and a decrease in call peak frequency and duration relative to pre-buzz calls, and resemble the terminal buzz phase calls of an aerial hawking bat's echolocation attack sequence. Sonar strobe groups (SSGs) are clustered sequences of non-buzz calls whose pulse intervals (PIs) are fairly regular and shorter than the PIs both before and after the cluster, but longer than the PIs of buzz calls. Like buzzes, SSGs are thought to indicate increased auditory attention. We recorded the echolocation calls emitted by juvenile big brown bats (Eptesicus fuscus) over postnatal development from birth to 32 days old, when full flight has normally been achieved, and tested the following hypotheses: (i) buzz production precedes the onset of controlled, powered flight; (ii) the emission of SSGs precedes buzzes and coincides with the onset of fluttering behaviour; and (iii) the onset of flight is attained first by young bats with adult-like wing loadings. We found that E. fuscus pups emitted landing buzzes before they achieved powered flight and produced SSGs several days before emitting landing buzzes. Both observations indicate that the onset of adult-like echolocation behaviour occurs prior to adult-like flight behaviour. Pups that achieved flight first were typically those that also first achieved low, adult-like wing loadings. Our results demonstrate that echolocation and flight develop in parallel but may be temporally offset, such that the sensory system precedes the locomotory system during postnatal ontogeny.

An experimental test of energy and electrolyte supplementation as a mitigation strategy for white-nose syndrome.

Fungi are increasingly recognised as harmful pathogens of wildlife. White-nose syndrome (WNS) is a fungal disease that has killed millions of hibernating bats in North America. High mortality has driven research to identify management strategies for the disease. Increased energy expenditure and fat depletion, as well as fluid loss, hypotonic dehydration and electrolyte depletion appear to be key aspects of WNS pathophysiology. Bats with WNS spend energy too quickly and also lose fluids containing water and electrolytes from lesions on exposed skin surfaces. During periodic arousals, bats often drink water but, in most of the WNS-affected area, food is not available during winter and, therefore, they cannot maintain energy balance or replace lost electrolytes. Therefore, providing a liquid caloric/electrolyte/nutrient supplement could be useful for treating WNS. We studied captive, hibernating little brown bats (Myotis lucifugus) to test whether providing supplemental energy and electrolytes (a 1:1 dilution of unflavoured Pedialyte) to hibernating bats could reduce severity of WNS symptoms and increase survival. Infected bats in the Pedialyte-supplemented group generally avoided the Pedialyte and preferentially drank plain water. We did not observe any differences in survival, arousal frequency or blood chemistry, but bats in the Pedialyte-supplemented group had higher fungal load and more UV fluorescence than the control group that was only provided with water. Supplemental electrolytes would be an attractive management strategy because of their low cost and logistic feasibility but our results suggest this approach would be ineffective. However, it could be useful to conduct preference experiments with multiple dilutions and/or flavours of electrolyte solution. Although they did not prefer Pedialyte in our experiment, bats in the hand readily drink it and electrolyte supplementation remains an important tool for rehabilitation of captive bats recovering from WNS and other causes of dehydration.

Body temperatures of hibernating little brown bats reveal pronounced behavioural activity during deep torpor and suggest a fever response during white-nose syndrome.

Hibernating animals use torpor [reduced body temperature (T b) and metabolic rate] to reduce energy expenditure during winter. Periodic arousals to normal T b are energetically expensive, so hibernators trade off arousal benefits against energetic costs. This is especially important for bats with white-nose syndrome (WNS), a fungal disease causing increased arousal frequency. Little brown bats (Myotis lucifugus) with WNS show upregulation of endogenous pyrogens and sickness behaviour. Therefore, we hypothesized that WNS should cause a fever response characterized by elevated T b. Hibernators could also accrue some benefits of arousals with minimal T b increase, thus avoiding full arousal costs. We compared skin temperature (T sk) of captive Myotis lucifugus inoculated with the WNS-causing fungus to T sk of sham-inoculated controls. Infected bats re-warmed to higher T sk during arousals which is consistent with a fever response. Torpid T sk did not differ. During what we term "cold arousals", bats exhibited movement following T sk increases of only 2.2 ± 0.3 °C, compared to >20 °C increases during normal arousals. Cold arousals occurred in both infected and control bats, suggesting they are not a pathophysiological consequence of WNS. Fever responses are energetically costly and could exacerbate energy limitation and premature fat depletion for bats with WNS. Cold arousals could represent an energy-saving mechanism for both healthy and WNS-affected bats when complete arousals are unnecessary or too costly. A few cold arousals were observed mid-hibernation, typically in response to disturbances. Cold arousals may, therefore, represent a voluntary restriction of arousal temperature instead of loss of thermoregulatory control.

White-nose syndrome increases torpid metabolic rate and evaporative water loss in hibernating bats.

Fungal diseases of wildlife typically manifest as superficial skin infections but can have devastating consequences for host physiology and survival. White-nose syndrome (WNS) is a fungal skin disease that has killed millions of hibernating bats in North America since 2007. Infection with the fungus Pseudogymnoascus destructans causes bats to rewarm too often during hibernation, but the cause of increased arousal rates remains unknown. On the basis of data from studies of captive and free-living bats, two mechanistic models have been proposed to explain disease processes in WNS. Key predictions of both models are that WNS-affected bats will show 1) higher metabolic rates during torpor (TMR) and 2) higher rates of evaporative water loss (EWL). We collected bats from a WNS-negative hibernaculum, inoculated one group with P. destructans, and sham-inoculated a second group as controls. After 4 mo of hibernation, TMR and EWL were measured using respirometry. Both predictions were supported, and our data suggest that infected bats were more affected by variation in ambient humidity than controls. Furthermore, disease severity, as indicated by the area of the wing with UV fluorescence, was positively correlated with EWL, but not TMR. Our results provide the first direct evidence that heightened energy expenditure during torpor and higher EWL independently contribute to WNS pathophysiology, with implications for the design of potential treatments for the disease.

Morphological, olfactory, and vocal development in big brown bats.

Using a within subjects design, we documented morphological, bioacoustical and behavioral developmental changes in big brown bats. Eptesicus fuscus pups are born naked and blind but assume an adult-like appearance by post-natal day (PND) 45 and flight by PND 30. Adult females use spatial memory, acoustic and olfactory cues to reunite with offspring, but it is unclear if pups can recognize maternal scents. We tested the olfactory discrimination abilities of young E. fuscus pups and found they exhibited no odor preferences. Pups also emit distinct vocalizations called isolation calls (i-calls) that facilitate mother-offspring reunions, but how pups shift their vocalizations from i-calls to downward frequency modulated (FM) sweeps used in echolocation remains unclear. Between PND 0-9, pups emitted mainly long duration, tonal i-calls rich in harmonics, but after they switched to short duration, downward FM sweeps with fewer harmonics. Call maximum frequency and repetition rate showed minor changes across development. Signal duration, bandwidth, and number of harmonics decreased, whereas the maximum, minimum and bandwidth of the fundamental, and peak spectral frequency all increased. We recorded vocalizations during prolonged maternal separation and found that isolated pups called longer and at a faster rate, presumably to signal for maternal assistance. To assess how PND 13 pups alter their signals during interactions with humans we compared spontaneous and provoked vocalizations and found that provoked calls were spectrally and temporally more similar to those of younger bats suggesting that pups in distress emit signals that sound like younger bats to promote maternal assistance.