A CUC1/auxin genetic module links cell polarity to patterned tissue growth and leaf shape diversity in crucifer plants.

How tissue-level information encoded by fields of regulatory gene activity is translated into the patterns of cell polarity and growth that generate the diverse shapes of different species remains poorly understood. Here, we investigate this problem in the case of leaf shape differences between Arabidopsis thaliana, which has simple leaves, and its relative Cardamine hirsuta that has complex leaves divided into leaflets. We show that patterned expression of the transcription factor CUP-SHAPED COTYLEDON1 in C. hirsuta (ChCUC1) is a key determinant of leaf shape differences between the two species. Through inducible genetic perturbations, time-lapse imaging of growth, and computational modeling, we find that ChCUC1 provides instructive input into auxin-based leaf margin patterning. This input arises via transcriptional regulation of multiple auxin homeostasis components, including direct activation of WAG kinases that are known to regulate the polarity of PIN-FORMED auxin transporters. Thus, we have uncovered a mechanism that bridges biological scales by linking spatially distributed and species-specific transcription factor expression to cell-level polarity and growth, to shape diverse leaf forms.

Echoes through time: amazing inferences from a fossil bat.

Sister to the Chiroptera crown-clade, the 50 million year old Vielasia sigei is suggested to have used laryngeal echolocation based on morphometric analyses. We discuss how Vielasia's discovery influences our understanding of the evolution of echolocation in bats and the insights fossils provide to the lives of extinct species.

Heterothermic Migration Strategies in Flying Vertebrates.

Migration is a widespread and highly variable trait among animals. Population-level patterns arise from individual-level decisions, including physiological and energetic constraints. Many aspects of migration are influenced by behaviors and strategies employed during periods of stopover, where migrants may encounter variable or unpredictable conditions. Thermoregulation can be a major cost for homeotherms which largely encounter ambient temperatures below the lower critical temperature during migration, especially during the rest phase of the daily cycle. In this review we describe the empirical evidence, theoretical models, and potential implications of bats and birds that use heterothermy to reduce thermoregulatory costs during migration. Torpor-assisted migration is a strategy described for migrating temperate insectivorous bats, whereby torpor can be used during periods of inactivity to drastically reduce thermoregulatory costs and increase net refueling rate, leading to shorter stopover duration, reduced fuel load requirement, and potential consequences for broad-scale movement patterns and survival. Hummingbirds can adopt a similar strategy, but most birds are not capable of torpor. However, there is an increasing recognition of the use of more shallow heterothermic strategies by diverse bird species during migration, with similarly important implications for migration energetics. A growing body of published literature and preliminary data from ongoing research indicate that heterothermic migration strategies in birds may be more common than traditionally appreciated. We further take a broad evolutionary perspective to consider heterothermy as an alternative to migration in some species, or as a conceptual link to consider alternatives to seasonal resource limitations. There is a growing body of evidence related to heterothermic migration strategies in bats and birds, but many important questions related to the broader implications of this strategy remain.

Lean Mass Dynamics in Hibernating Bats and Implications for Energy and Water Budgets.

AbstractHibernation requires balancing energy and water demands over several months. Many studies have noted the importance of fat for hibernation energy budgets, but protein catabolism in hibernation has received less attention, and whole-animal changes in lean mass have not previously been considered. We used quantitative magnetic resonance body composition analysis to measure deposition of fat and lean mass of cave myotis (Myotis velifer) during the prehibernation period and decreases in fat and lean mass of Townsend's big-eared bats (Corynorhinus townsendii) during hibernation. For cave myotis, lean mass represented 25% and 38% (female and male, respectively) of prehibernation mass gain. In hibernating Townsend's big-eared bats, lean mass decrease was similar for females and males. We used values for Townsend's big-eared bats to explore the functional implications of lean mass change for water and energy budgets. Lean mass accounted for a substantial proportion of mass change during hibernation (female: 18%, male: 35%), and although not accounting for a large proportion of the energy budget (female: 3%, male: 7%), lean mass catabolism represented an important contribution to water production (female: 14%, male: 29%). Although most mammals cannot rely on protein catabolism for metabolic water production because of the water cost of excreting urea, we propose a variation of the protein-for-water strategy whereby hibernators could temporally compartmentalize the benefits of protein catabolism to periods of torpor and the water cost to periodic arousals when free drinking water is typically available. Combined, our analyses demonstrate the importance of considering changes in lean mass during hibernation.

Relating wing morphology and immune function to patterns of partial and differential bat migration using stable isotopes.

Migration is energetically expensive and is predicted to drive similar morphological adaptations and physiological trade-offs in migratory bats and birds. Previous studies suggest that fixed traits like wing morphology vary among species and individuals according to selective pressures on flight, while immune defences can vary flexibly within individuals as energy is variably reallocated throughout the year. We assessed intraspecific variation in wing morphology and immune function in silver-haired bats Lasionycteris noctivagans, a species that follows both partial and differential migration patterns. We hypothesized that if bats experience energy constraints associated with migration, then wing morphology and immune function should vary based on migratory tendency (sedentary or migratory) and migration distance. We predicted that long-distance migrants would have reduced immune function and more migration-adapted wing shapes compared to resident or short-distance migrating bats. We estimated breeding latitude of spring migrants using stable hydrogen isotope techniques. Our sample consisted primarily of male bats, which we categorized as residents, long-distance northern migrants, short-distance northern migrants and southern migrants (apparent breeding location south of capture site). Controlling for individual condition and capture date, we related wing characteristics and immune indices among groups. Some, but not all, aspects of wing form and immune function varied between migrants and residents. Long-distance northern migrants had larger wings than short-distance northern migrants and lower wing loading than southern migrants. Compared with resident bats, short-distance northern migrants had reduced IgG while southern migrants had heightened neutrophils and neutrophil-to-lymphocyte ratios. Body fat, aspect ratio, wing tip shape and bacteria killing ability did not vary with migration status or distance. In general, male silver-haired bats do not appear to mediate migration costs by substantially downregulating immune defences or to be under stronger selection for wing forms adapted for fast, energy-efficient flight. Such phenotypic changes may be more adaptive for female silver-haired bats, which migrate farther and are more constrained by time in spring than males. Adaptations for aerial hawking and the use of heterothermy by migrating bats may also reduce the energetic cost of migration and the need for more substantial morphological and physiological trade-offs.

Experimental inoculation trial to determine the effects of temperature and humidity on White-nose Syndrome in hibernating bats.

Disease results from interactions among the host, pathogen, and environment. Inoculation trials can quantify interactions among these players and explain aspects of disease ecology to inform management in variable and dynamic natural environments. White-nose Syndrome, a disease caused by the fungal pathogen, Pseudogymnoascus destructans (Pd), has caused severe population declines of several bat species in North America. We conducted the first experimental infection trial on the tri-colored bat, Perimyotis subflavus, to test the effect of temperature and humidity on disease severity. We also tested the effects of temperature and humidity on fungal growth and persistence on substrates. Unexpectedly, only 37% (35/95) of bats experimentally inoculated with Pd at the start of the experiment showed any infection response or disease symptoms after 83 days of captive hibernation. There was no evidence that temperature or humidity influenced infection response. Temperature had a strong effect on fungal growth on media plates, but the influence of humidity was more variable and uncertain. Designing laboratory studies to maximize research outcomes would be beneficial given the high costs of such efforts and potential for unexpected outcomes. Understanding the influence of microclimates on host-pathogen interactions remains an important consideration for managing wildlife diseases, particularly in variable environments.

Increasing understanding of environmental modifications using the Virtual Dementia Experience for professional carers of people living with dementia.

Modifications to the environment can have a positive impact on the person living with dementia. The purpose of this study was to examine the effects of a virtual reality (VR) educational workshop on improving understanding of the impact of environments for people living with dementia for carers in residential aged care facilities in Australia. Participants were nonrandomly allocated to one of two conditions: a VR condition which enabled them to take the perspective of a person living with dementia as they navigate a home environment (n = 40), or to a non-VR condition, in which participants received equivalent information in a classroom without the use of VR (n = 56). Participants completed a purpose-built pictorial measure of their understanding of environmental factors that could impact on people with dementia before and after the workshop. Following education, an analysis of covariance revealed that compared to non-VR education, those who received VR education identified a significantly greater number of appropriate environmental modifications, after adjusting for practice test scores. The results from this study demonstrate the utility of VR as a promising tool to improve carers' understanding of the ways in which the environment can be modified to assist people living with dementia.

Similar hibernation physiology in bats across broad geographic ranges.

Species with broad geographic ranges may experience varied environmental conditions throughout their range leading to local adaptation. Variation among populations reflects potential adaptability or plasticity, with implications for populations impacted by disease, climate change, and other anthropogenic influences. However, behavior may counteract divergent selection among populations. We studied intraspecific variation in hibernation physiology of Myotis lucifugus (little brown myotis) and Corynorhinus townsendii (Townsend's big-eared bat), two species of bats with large geographic ranges. We studied M. lucifugus at three hibernacula which spanned a latitudinal gradient of 1500 km, and C. townsendii from 6 hibernacula spread across 1200 km latitude and 1200 km longitude. We found no difference in torpid metabolic rate among populations of either species, nor was there a difference in the effect of ambient temperature among sites. Evaporative water loss was similar among populations of both species, with the exception of one C. townsendii pairwise site difference and one M. lucifugus site that differed from the others. We suggest the general lack of geographic variation is a consequence of behavioral microhabitat selection. As volant animals, bats can travel relatively long distances in search of preferred microclimates for hibernation. Despite dramatic macroclimate differences among populations, hibernating bats are able to find preferred microclimate conditions within their range, resulting in similar selection pressures among populations spread across wide geographic ranges.

Interspecific variation in evaporative water loss and temperature response, but not metabolic rate, among hibernating bats.

Hibernation is widespread among mammals in a variety of environmental contexts. However, few experimental studies consider interspecific comparisons, which may provide insight into general patterns of hibernation strategies. We studied 13 species of free-living bats, including populations spread over thousands of kilometers and diverse habitats. We measured torpid metabolic rate (TMR) and evaporative water loss (two key parameters for understanding hibernation energetics) across a range of temperatures. There was no difference in minimum TMR among species (i.e., all species achieved similarly low torpid metabolic rate) but the temperature associated with minimum TMR varied among species. The minimum defended temperature (temperature below which TMR increased) varied from 8 °C to < 2 °C among species. Conversely, evaporative water loss varied among species, with species clustered in two groups representing high and low evaporative water loss. Notably, species that have suffered population declines due to white-nose syndrome fall in the high evaporative water loss group and less affected species in the low evaporative water loss group. Documenting general patterns of physiological diversity, and associated ecological implications, contributes to broader understanding of biodiversity, and may help predict which species are at greater risk of environmental and anthropogenic stressors.

What is winter? Modeling spatial variation in bat host traits and hibernation and their implications for overwintering energetics.

White-nose syndrome (WNS) has decimated hibernating bat populations across eastern and central North America for over a decade. Disease severity is driven by the interaction between bat characteristics, the cold-loving fungal agent, and the hibernation environment. While we further improve hibernation energetics models, we have yet to examine how spatial heterogeneity in host traits is linked to survival in this disease system. Here, we develop predictive spatial models of body mass for the little brown myotis (Myotis lucifugus) and reassess previous definitions of the duration of hibernation of this species. Using data from published literature, public databases, local experts, and our own fieldwork, we fit a series of generalized linear models with hypothesized abiotic drivers to create distribution-wide predictions of prehibernation body fat and hibernation duration. Our results provide improved estimations of hibernation duration and identify a scaling relationship between body mass and body fat; this relationship allows for the first continuous estimates of prehibernation body mass and fat across the species' distribution. We used these results to inform a hibernation energetic model to create spatially varying fat use estimates for M. lucifugus. These results predict WNS mortality of M. lucifugus populations in western North America may be comparable to the substantial die-off observed in eastern and central populations.

Temperature alone is insufficient to understand hibernation energetics.

Energy conservation has long been a focal point in hibernation research. A long-standing assumption is that ambient temperature (Ta) largely defines the rate of energy expenditure because of well-known relationships between Ta, metabolic rate and frequency of arousal from torpor. Body condition and humidity also affect energy expenditure but are usually considered secondary factors. We held tricolored bats (Perimyotis subflavus) in captivity under multiple environmental conditions to directly compare the importance of Ta, fat mass and humidity for hibernation energy expenditure. Fat mass was the best predictor of female mass loss, followed by Ta and humidity. However, males had less fat and adopted a more energetically conservative hibernation strategy. Our results demonstrate that understanding the evolution of behavior, physiology and ecology of hibernation requires disentangling the relative contributions of multiple drivers of hibernation energetics, and that Ta is not always the most important factor driving energy expenditure.

Exploiting common senses: sensory ecology meets wildlife conservation and management.

Multidisciplinary approaches to conservation and wildlife management are often effective in addressing complex, multi-factor problems. Emerging fields such as conservation physiology and conservation behaviour can provide innovative solutions and management strategies for target species and systems. Sensory ecology combines the study of 'how animals acquire' and process sensory stimuli from their environments, and the ecological and evolutionary significance of 'how animals respond' to this information. We review the benefits that sensory ecology can bring to wildlife conservation and management by discussing case studies across major taxa and sensory modalities. Conservation practices informed by a sensory ecology approach include the amelioration of sensory traps, control of invasive species, reduction of human-wildlife conflicts and relocation and establishment of new populations of endangered species. We illustrate that sensory ecology can facilitate the understanding of mechanistic ecological and physiological explanations underlying particular conservation issues and also can help develop innovative solutions to ameliorate conservation problems.

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.

Incorporating evaporative water loss into bioenergetic models of hibernation to test for relative influence of host and pathogen traits on white-nose syndrome.

Hibernation consists of extended durations of torpor interrupted by periodic arousals. The 'dehydration hypothesis' proposes that hibernating mammals arouse to replenish water lost through evaporation during torpor. Arousals are energetically expensive, and increased arousal frequency can alter survival throughout hibernation. Yet we lack a means to assess the effect of evaporative water loss (EWL), determined by animal physiology and hibernation microclimate, on torpor bout duration and subsequent survival. White-nose syndrome (WNS), a devastating disease impacting hibernating bats, causes increased frequency of arousals during hibernation and EWL has been hypothesized to contribute to this increased arousal frequency. WNS is caused by a fungus, which grows well in humid hibernaculum environments and damages wing tissue important for water conservation. Here, we integrated the effect of EWL on torpor expression in a hibernation energetics model, including the effects of fungal infection, to determine the link between EWL and survival. We collected field data for Myotis lucifugus, a species that experiences high mortality from WNS, to gather parameters for the model. In saturating conditions, we predicted healthy bats experience minimal mortality. Infected bats, however, suffer high fungal growth in highly saturated environments, leading to exhaustion of fat stores before spring. Our results suggest that host adaptation to humid environments leads to increased arousal frequency from infection, which drives mortality across hibernaculum conditions. Our modified hibernation model provides a tool to assess the interplay between host physiology, hibernaculum microclimate, and diseases such as WNS on winter survival.

Migration and reproduction are associated with similar degrees of phenotypic flexibility in an insectivorous bat.

As organisms face variation in energetic challenges and physiological demands, they often respond with reversible changes in behavior, physiology, and morphology, described as phenotypic flexibility. From the magnitude of phenotypic change, we can infer the energetic challenges of different life stages. We studied phenotypic flexibility in a population of reproductive and pre-migratory female insectivorous bats (Tadarida brasiliensis). While female reproductive demands are well described in insectivorous bats, there are questions regarding the demands of migration. Our objective was to measure phenotypic flexibility to assess the cost of autumn migration compared to reproduction in an insectivorous bat. We measured plasma triglycerides to quantify foraging rate, and body composition (body mass and individual organ mass) of T. brasiliensis throughout the summer season (from arrival in spring through pre-migration/migration departure in autumn) according to the female reproductive cycle. We found phenotypic changes during pre-migration/migration similar to periods of high-energy demand during reproduction (e.g., late pregnancy and lactation). Most notably, bats weighed as much during peak pregnancy, as they did during migration, and the rapid mass gain from post-lactation through the migratory period was due to a combination of hyperphagia and hypertrophy of digestive organs. Our results indicate that energetic demands incurred during migration are similar to those during reproduction and emphasize the energetic challenges of migration.

Seasonal Dynamics of Lipid Metabolism and Energy Storage in the Brazilian Free-Tailed Bat.

As small, flying, mammalian endotherms, insectivorous bats are adapted to operate at high levels of energy expenditure. In response to seasonally variable challenges, we predicted that bats should balance energy budgets by flexibly adjusting aspects of their physiology or behavior in ways that elevate metabolic capacity. We examined variation in energy storage and pathways for oxidative metabolism in Brazilian free-tailed bats (Tadarida brasiliensis) related to estimated costs associated with reproduction and migration. We collected pectoral muscle and liver from female T. brasiliensis at six time points during the summer and fall and measured changes in the activity of four enzymes involved with lipid metabolism. Body mass varied substantially with life-cycle stage, suggesting that rapid accumulation and use of fat stores occurs in response to current and anticipated energy demands. Catabolic enzyme activity (carnitine palmitoyl transferase [CPT], 3-hydroxyacyl-CoA dehydrogenase [HOAD], and citrate synthase [CS]) in the muscle was increased during lactation compared with early pregnancy but exhibited no change before fall migration. While there was no temporal change in lipid biosynthetic capacity in the liver, fatty acid synthase activity was negatively correlated with body mass. Variation in body mass and enzyme activity in T. brasiliensis during the summer suggests that stored energy is mobilized and lipid oxidative capacity is increased during periods of increased demand and that lipid biosynthetic capacity is increased with depletion of fat stores. These results suggest that bats are able to flexibly adjust metabolic capacity based on energy requirement to maintain energy balance despite high levels of expenditure.

Testing the "Fasting While Foraging" Hypothesis: Effects of Recent Feeding on Plasma Metabolite Concentrations in Little Brown Bats (Myotis lucifugus).

Plasma metabolite concentrations can be used to understand nutritional status and foraging behavior across ecological contexts including prehibernation fattening, migration refueling, and variation in foraging habitat quality. Generally, high plasma concentrations of the ketone ß-hydroxybutyrate, a product of fat catabolism, indicate fasting, while triglycerides indicate recent feeding and fat accumulation. In recent studies of insectivorous bats, triglyceride concentration increased after feeding as expected, but ß-hydroxybutyrate also unexpectedly increased rather than decreased. An aerial-hawking foraging strategy is energetically demanding, and thus it has been hypothesized that foraging by insectivorous bats requires catabolism of stored fat. We tested this hypothesis by quantifying plasma ß-hydroxybutyrate and triglyceride concentration following feeding in little brown bats (Myotis lucifugus) that were temporarily housed in individual cages to prevent flight. We provided a fixed amount of food and collected blood samples at different intervals after feeding to produce variation in plasma metabolite concentrations. Plasma triglyceride concentration responded as predicted, but similar to previous studies and contrary to our prediction, when flight was eliminated plasma ß-hydroxybutyrate concentration responded similarly to triglyceride. Thus, it is unlikely that the unexpected plasma ß-hydroxybutyrate patterns observed in previous studies were related to flight. The mechanism underlying this unexpected pattern remains unknown, but the response has been consistent in all studies to date. Thus, plasma metabolite analysis provides an effective tool for studies of nutritional status, although more work is needed to understand why insectivorous bats respond differently than other taxa.

Bats are not squirrels: Revisiting the cost of cooling in hibernating mammals.

Many species use stored energy to hibernate through periods of resource limitation. Hibernation, a physiological state characterized by depressed metabolism and body temperature, is critical to winter survival and reproduction, and therefore has been extensively quantified and modeled. Hibernation consists of alternating phases of extended periods of torpor (low body temperature, low metabolic rate), and energetically costly periodic arousals to normal body temperature. Arousals consist of multiple phases: warming, euthermia, and cooling. Warming and euthermic costs are regularly included in energetic models, but although cooling to torpid body temperature is an important phase of the torpor-arousal cycle, it is often overlooked in energetic models. When included, cooling cost is assumed to be 67% of warming cost, an assumption originally derived from a single study that measured cooling cost in ground squirrels. Since this study, the same proportional value has been assumed across a variety of hibernating species. However, no additional values have been derived. We derived a model of cooling cost from first principles and validated the model with empirical energetic measurements. We compared the assumed 67% proportional cooling cost with our model-predicted cooling cost for 53 hibernating mammals. Our results indicate that using 67% of warming cost only adequately represents cooling cost in ground squirrel-sized mammals. In smaller species, this value overestimates cooling cost and in larger species, the value underestimates cooling cost. Our model allows for the generalization of energetic costs for multiple species using species-specific physiological and morphometric parameters, and for predictions over variable environmental conditions.

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.

Environmentally persistent pathogens present unique challenges for studies of host-pathogen interactions: Reply to Field (2018).

Linked article: https://doi.org/10.1002/ece3.4034.