Biomarker Metabolites Discriminate between Physiological States of Field, Cave and White-nose Syndrome Diseased Bats.

Analysis of volatile organic compound (VOC) emissions using electronic-nose (e-nose) devices has shown promise for early detection of white-nose syndrome (WNS) in bats. Tricolored bats, Perimyotis subflavus, from three separate sampling groups defined by environmental conditions, levels of physical activity, and WNS-disease status were captured temporarily for collection of VOC emissions to determine relationships between these combinations of factors and physiological states, Pseudogymnoascus destructans (Pd)-infection status, and metabolic conditions. Physiologically active (non-torpid) healthy individuals were captured outside of caves in Arkansas and Louisiana. In addition, healthy and WNS-diseased torpid bats were sampled within caves in Arkansas. Whole-body VOC emissions from bats were collected using portable air-collection and sampling-chamber devices in tandem. Electronic aroma-detection data using three-dimensional Principal Component Analysis provided strong evidence that the three groups of bats had significantly different e-nose aroma signatures, indicative of different VOC profiles. This was confirmed by differences in peak numbers, peak areas, and tentative chemical identities indicated by chromatograms from dual-column GC-analyses. The numbers and quantities of VOCs present in whole-body emissions from physiologically active healthy field bats were significantly greater than those of torpid healthy and diseased cave bats. Specific VOCs were identified as chemical biomarkers of healthy and diseased states, environmental conditions (outside and inside of caves), and levels of physiological activity. These results suggest that GC/E-nose dual-technologies based on VOC-detection and analyses of physiological states, provide noninvasive alternative means for early assessments of Pd-infection, WNS-disease status, and other physiological states.

Assessment of the Portable C-320 Electronic Nose for Discrimination of Nine Insectivorous Bat Species: Implications for Monitoring White-Nose Syndrome.

The development of new C-320 electronic-nose (e-nose) methods for pre-symptomatic detection of White-Nose Syndrome (WNS) in bats has required efficacy studies of instrument capabilities to discriminate between major sources of volatile organic compounds (VOCs) derived from clinical samples. In this phase-2 study, we further tested this e-nose for capabilities to distinguish between bat species based on differences in whole-body VOC emissions. Live healthy individuals of nine bat species were temporarily captured outside of caves in Arkansas and Louisiana. VOC emissions from bats were collected using newly developed portable air collection and sampling-chamber devices in tandem. Sensor-array output responses to bat VOC emissions were compared to those of 22 pure VOC analytical standards from five chemical classes. Distinct smellprint signatures were produced from e-nose analyses of VOC metabolites derived from individual bat species. Smellprint patterns were analyzed using 2-dimensional and 3-dimensional Principal Component Analysis (PCA) to produce aroma map plots showing effective discrimination between bat species with high statistical significance. These results demonstrate potential instrument efficacy for distinguishing between species-specific, bat-derived VOC metabolite emissions as major components of clinical samples collected from bats in caves for disease detection prior to symptom development. This study provided additional information required to fully test the efficacy of a portable e-nose instrument for diagnostic applications in subsequent phase-3 testing of noninvasive, early WNS disease detection in intra-cave hibernating bats.

A physiological understanding of organismal responses to fire.

Devastation of both natural and human habitats due to wildfires is becoming an increasingly prevalent global issue. Fire-adapted and fire-prone regions, such as California and parts of Australia, are experiencing more frequent and increasingly destructive wildfires, accompanied by longer wildfire seasons. Further, wildfires are becoming more commonplace in areas that historically do not regularly experience fire, causing an increased risk of habitat loss in less resilient ecosystems. The escalation of fire outbreaks is a result of several factors; however, at the forefront of these outbreaks is an increase in highly flammable dry vegetation due to sustained drought, a trend we will see growing in our changing climate. To mitigate the potentially detrimental outcomes of wildfires, it is imperative that we understand the response of ecosystems to fire not only from an ecological perspective, but also from a physiological perspective. Research focused on the physiological adaptations of organisms to environmental constraints caused by fire can give insight into how plants and animals respond to fire, on both short- and long-term scales. Importantly, this information needs to be adapted effectively into fire management plans to improve the recovery success of organisms after fire.

A burning question: what are the risks and benefits of mammalian torpor during and after fires?

Although wildfires are increasing globally, available information on how mammals respond behaviourally and physiologically to fires is scant. Despite a large number of ecological studies, often examining animal diversity and abundance before and after fires, the reasons as to why some species perform better than others remain obscure. We examine how especially small mammals, which generally have high rates of energy expenditure and food requirements, deal with fires and post-fire conditions. We evaluate whether mammalian torpor, characterised by substantial reductions in body temperature, metabolic rate and water loss, plays a functional role in survival of mammals impacted by fires. Importantly, torpor permits small mammals to reduce their activity and foraging, and to survive on limited food. Torpid small mammals (marsupials and bats) can respond to smoke and arouse from torpor, which provides them with the possibility to evade direct exposure to fire, although their response is often slowed when ambient temperature is low. Post-fire conditions increase expression of torpor with a concomitant decrease in activity for free-ranging echidnas and small forest-dwelling marsupials, in response to reduced cover and reduced availability of terrestrial insects. Presence of charcoal and ash increases torpor use by captive small marsupials beyond food restriction alone, likely in anticipation of detrimental post-fire conditions. Interestingly, although volant bats use torpor on every day after fires, they respond by decreasing torpor duration, and increasing activity, perhaps because of the decrease in clutter and increase in foraging opportunities due to an increase in aerial insects. Our summary shows that torpor is an important tool for post-fire survival and, although the physiological and behavioural responses of small mammals to fire are complex, they seem to reflect energetic requirements and mode of foraging. We make recommendations on the conditions during management burns that are least likely to impact heterothermic mammals.

Can bats sense smoke during deep torpor?

While torpor is a beneficial energy-saving strategy, it may incur costs if an animal is unable to respond appropriately to external stimuli, which is particularly true when it is necessary to escape from threats such as fire. We aimed to determine whether torpid bats, which are potentially threatened because they must fly to escape, can sense smoke and whether respiration rate (RR), heart rate (HR) and reaction time of torpid bats prior to and following smoke introduction is temperature-dependent. To test this we quantified RR and HR of captive Australian tree-roosting bats, Nyctophilus gouldi (n=5, ~10g), in steady-state torpor in response to short-term exposure to smoke from Eucalyptus spp. leaves between ambient temperatures (Ta) of 11 and 23°C. Bats at lower Ta took significantly longer (28-fold) to respond to smoke, indicated by a cessation of episodic breathing and a rapid increase in RR. Bats at lower Ta returned to torpor more swiftly following smoke exposure than bats at higher Ta. Interestingly, bats at Ta<15°C never returned to thermoconforming steady-state torpor prior to the end of the experimental day, whereas all bats at Ta≥15°C did, as indicated by apnoeic HR. This shows that although bats at lower Ta took longer to respond, they appear to maintain vigilance and prevent deep torpor after the first smoke exposure, likely to enable fast escape. Our study reveals that bats can respond to smoke stimuli while in deep torpor. These results are particularly vital within the framework of fire management conducted at Ta<15°C, as most management burns are undertaken during winter when bats will likely respond more slowly to fire cues such as smoke, delaying the time to escape from the fire.

Black or white? Physiological implications of roost colour and choice in a microbat.

Although roost choice in bats has been studied previously, little is known about how opposing roost colours affect the expression of torpor quantitatively. We quantified roost selection and thermoregulation in a captive Australian insectivorous bat, Nyctophilus gouldi (n=12) in winter when roosting in black and white coloured boxes using temperature-telemetry. We quantified how roost choice influences torpor expression when food was provided ad libitum or restricted in bats housed together in an outdoor aviary exposed to natural fluctuations of ambient temperature. Black box temperatures averaged 5.1°C (maximum 7.5°C) warmer than white boxes at their maximum daytime temperature. Bats fed ad libitum chose black boxes on most nights (92.9%) and on 100% of nights when food-restricted. All bats used torpor on all study days. However, bats fed ad libitum and roosting in black boxes used shorter torpor and spent more time normothermic/active at night than food-restricted bats and bats roosting in white boxes. Bats roosting in black boxes also rewarmed passively more often and to a higher skin temperature than those in white boxes. Our study suggests that N. gouldi fed ad libitum select warmer roosts in order to passively rewarm to a higher skin temperature and thus save energy required for active midday rewarming as well as to maintain a normothermic body temperature for longer periods at night. This study shows that colour should be considered when deploying bat boxes; black boxes are preferable for those bats that use passive rewarming, even in winter when food availability is reduced.

Post-wildfire physiological ecology of an Australian microbat.

Historical patterns of wildfires are being altered as a result of changing climate and therefore are becoming an increasingly pressing global issue. How small mammals deal physiologically with changes in landscape and food availability due to fire remains largely unknown, although recent studies on small heterothermic terrestrial mammals have shown an increase in post-fire torpor use to reduce energy and foraging requirements. However, data on the behavioural and physiological responses of bats after fires are scarce, although potentially these volant species may differ from terrestrial mammals. Therefore, we investigated the post-fire thermal biology and activity of lesser long-eared bats (Nyctophilus geoffroyi) using temperature-telemetry in Warrumbungle National Park, NSW, which experienced a devastating wildfire in 2013. The study comprised two field seasons, one in 2013 within 4 months after the fire, and one in 2015 two years after the fire to identify potential changes in behaviour and physiology. Interestingly, soon after the fire, bats showed significantly shorter torpor bout duration (11.8 ± 12.5 h) and longer normothermia duration (8.7 ± 4.6 h) in comparison to those in 2015 (torpor bout duration: 24.1 ± 23.5 h; normothermia duration: 2.5 ± 1.5 h). Insect availability was significantly (20-fold) higher in 2013 than in 2015, which was likely an important factor resulting in the short average torpor bout duration by N. geoffroyi after the fire. Our data indicate that volant bats appear to show the opposite post-fire behavioural and physiological responses to small terrestrial mammals, showing longer normothermic and active periods and shorter torpor bouts to capitalise on an increase in available post-fire resources.