Jonathan D. Reichard

Grand Challenges in Migration Biology

Abstract Billions of animals migrate each year. To successfully reach their destination, migrants must have evolved an appropriate genetic program and suitable developmental, morphological, physiological, biomechanical, behavioral, and life-history traits. Moreover, they must interact successfully with biotic and abiotic factors in their environment. Migration therefore provides an excellent model system in which to address several of the “grand challenges” in organismal biology. Previous research on migration, however, has often focused on a single aspect of the phenomenon, largely due to methodological, geographical, or financial constraints. Integrative migration biology asks ‘big questions’ such as how, when, where, and why animals migrate, which can be answered by examining the process from multiple ecological and evolutionary perspectives, incorporating multifaceted knowledge from various other scientific disciplines, and using new technologies and modeling approaches, all within the context of an annual cycle. Adopting an integrative research strategy will provide a better understanding of the interactions between biological levels of organization, of what role migrants play in disease transmission, and of how to conserve migrants and the habitats upon which they depend.

THERMAL IMAGING REVEALS SIGNIFICANTLY SMALLER BRAZILIAN FREE-TAILED BAT COLONIES THAN PREVIOUSLY ESTIMATED

Abstract Using data collected with thermal imaging technology, we found a major reduction in population estimates of colony size in the Brazilian free-tailed bat (Tadarida brasiliensis) from 54 million, obtained in 1957 without this technology, to 4 million in 6 major cave colonies in the southwestern United States. The 1957 census was based on human visual observations of cave emergence flights that were subject to potentially high errors. The recent census was produced using an accurate, reproducible counting method and based on complete temporal records of colony emergences. Analysis of emergence flights from dusk through darkness also revealed patterns in group behavior that would be difficult to capture without thermal infrared technology. Flow patterns of bats during emergence flights exhibited characteristic single, double, or triple episodes, with the peak flow during the 1st episode. A consistent rhythmic pattern of flow episodes and pauses was revealed across colonies and was independent of emergence tempo.

White-Nose Syndrome Inflicts Lasting Injuries to the Wings of Little Brown Myotis (Myotis lucifugus)

White-nose syndrome (WNS) is an emerging disease causing massive mortality of hibernating bats in the northeastern United States. At hibernacula, bats affected with WNS typically exhibit growth of a white psychrophylic fungus (Geomyces destructans) on the nose, wings and ears; many individuals seem to prematurely die of starvation owing to depleted fat reserves. Conspicuous scarring and necrosis of the wings on WNS-affected bats that survive hibernation may have lasting consequences for survival and reproductive success during the active season. We monitored two maternity colonies of little brown myotis, Myotis lucifugus, in Massachusetts and New Hampshire from 14 May to 8 August 2008 to assess body conditions after expected exposure to WNS over the previous winter. We developed a 4-point wing damage index (WDI = 0 to 3) to assess the incidence and severity of wing damage in the months following emergence from hibernation. Severe wing damage was observed up to 4 June and moderate damage was observed through 9 July. Light wing damage was observed on both adult and juvenile bats throughout the study period, but was not exclusively attributed to WNS. The most severe wing damage was associated with a lower body mass index which may reflect reduced foraging success. Overall, reproductive rate was 85.1% in 2008; slightly lower than reported in previous studies. The incidence, timing, and geographic range of wing damage observed on little brown myotis in 2008 correspond to the occurrence of WNS at hibernacula. Monitoring wing conditions of affected and healthy bats will be important tool for assessing the spread of this disease and for establishing baseline data for unaffected bats. The simple scale we propose should be useful for monitoring wing conditions in any bat species.

Specific alterations in complement protein activity of little brown myotis (Myotis lucifugus) hibernating in White-nose syndrome affected sites

White-nose syndrome (WNS) is the most devastating condition ever reported for hibernating bats, causing widespread mortality in the northeastern United States. The syndrome is characterized by cutaneous lesions caused by a recently identified psychrophilic and keratinophylic fungus (Geomyces destructans), depleted fat reserves, atypical behavior, and damage to wings; however, the proximate cause of mortality is still uncertain. To assess relative levels of immunocompetence in bats hibernating in WNS-affected sites compared with levels in unaffected bats, we describe blood plasma complement protein activity in hibernating little brown myotis (Myotis lucifugus) based on microbicidal competence assays using Escherichia coli, Staphylococcus aureus and Candida albicans. Blood plasma from bats collected during mid-hibernation at WNS-affected sites had higher bactericidal ability against E. coli and S. aureus, but lower fungicidal ability against C. albicans when compared with blood plasma from bats collected at unaffected sites. Within affected sites during mid-hibernation, we observed no difference in microbicidal ability between bats displaying obvious fungal infections compared to those without. Bactericidal ability against E. coli decreased significantly as hibernation progressed in bats collected from an affected site. Bactericidal ability against E. coli and fungicidal ability against C. albicans were positively correlated with body mass index (BMI) during late hibernation. We also compared complement activity against the three microbes within individuals and found that the ability of blood plasma from hibernating M. lucifugus to lyse microbial cells differed as follows: E. coli>S. aureus>C. albicans. Overall, bats affected by WNS experience both relatively elevated and reduced innate immune responses depending on the microbe tested, although the cause of observed immunological changes remains unknown. Additionally, considerable trade-offs may exist between energy conservation and immunological responses. Relationships between immune activity and torpor, including associated energy expenditure, are likely critical components in the development of WNS.

Hibernating Little Brown Myotis (Myotis lucifugus) Show Variable Immunological Responses to White-Nose Syndrome

White-nose syndrome (WNS) is an emerging infectious disease devastating hibernating North American bat populations that is caused by the psychrophilic fungus Geomyces destructans. Previous histopathological analysis demonstrated little evidence of inflammatory responses in infected bats, however few studies have compared other aspects of immune function between WNS-affected and unaffected bats. We collected bats from confirmed WNS-affected and unaffected sites during the winter of 2008–2009 and compared estimates of their circulating levels of total leukocytes, total immunoglobulins, cytokines and total antioxidants. Bats from affected and unaffected sites did not differ in their total circulating immunoglobulin levels, but significantly higher leukocyte counts were observed in bats from affected sites and particularly in affected bats with elevated body temperatures (above 20°C). Bats from WNS-affected sites exhibited significantly lower antioxidant activity and levels of interleukin-4 (IL-4), a cytokine that induces T cell differentiation. Within affected sites only, bats exhibiting visible fungal infections had significantly lower antioxidant activity and levels of IL-4 compared to bats without visible fungal infections. Overall, bats hibernating in WNS-affected sites showed immunological changes that may be evident of attempted defense against G. destructans. Observed changes, specifically elevated circulating leukocytes, may also be related to the documented changes in thermoregulatory behaviors of affected bats (i.e. increased frequencies in arousal from torpor). Alterations in immune function may reflect expensive energetic costs associated with these processes and intrinsic qualities of the immunocapability of hibernating bats to clear fungal infections. Additionally, lowered antioxidant activity indicates a possible imbalance in the pro- versus antioxidant system, may reflect oxidative tissue damage, and should be investigated as a contributor to WNS-associated morbidity and mortality.

Evening Emergence Behavior and Seasonal Dynamics in Large Colonies of Brazilian Free-Tailed Bats

Abstract Flexible behaviors permit gregarious animals to exploit spatially and temporally favorable conditions for reproduction. Evening emergences of Brazilian free-tailed bats (Tadarida brasiliensis) in south-central Texas were recorded to determine changes in colony dynamics and to evaluate hypotheses associated with predator avoidance and seasonal energy demands. In 2007, median parturition date was 18 June and lactation lasted about 54 days. Juveniles commenced flight after 41 days, then continued to suckle for 2 weeks before being weaned. Onset of evening emergence was correlated with sunset, beginning, on average, 11.8 min after sunset (95% confidence interval  =  7.1, 16.6), which is later than reported in previous studies. Pregnant females emerged later than their roost mates, consistent with a predator avoidance hypothesis. Lactating females emerged after sunset, but earlier than other bats, consistent with the hypothesis that this is a period of high energy demand. Juveniles emerged later than adult females while they continued to be suckled, but emerged earlier as they matured and were weaned, suggesting a trade-off between predator avoidance and energy demands. Our observations indicate that evening emergence behavior of T. brasiliensis varies with environmental cues but is mediated by the energetic demands of reproduction.

Thermoregulation during Flight: Body Temperature and Sensible Heat Transfer in Free-Ranging Brazilian Free-Tailed Bats (Tadarida brasiliensis)

Bat wings are important for thermoregulation, but their role in heat balance during flight is largely unknown. More than 80% of the energy consumed during flight generates heat as a by-product, and thus it is expected that bat wings should dissipate large amounts of heat to prevent hyperthermia. We measured rectal (Tr) and surface (Ts) temperatures of Brazilian free-tailed bats (Tadarida brasiliensis) as they emerged from and returned to their daytime roosts and calculated sensible heat transfer for different body regions (head, body, wings, and tail membrane). Bats’ Tr decreased from 36.8°C during emergence flights to 34.4°C during returns, and Ts scaled positively with ambient temperature (Ta). Total radiative heat loss from bats was significantly greater for a radiative sink to the night sky than for a sink with temperature equal to Ta. We found that free-ranging Brazilian free-tailed bats, on average, do not dissipate heat from their wings by convection but instead dissipate radiative heat (L) to the cloudless night sky during flight ( W). However, within the range of Ta measured in this study, T. brasiliensis experienced net heat loss between evening emergence and return flights. Regional hypothermia reduces heat loss from wings that are exposed to potentially high convective fluxes. Additional research is needed to establish the role of wings in evaporative cooling during flight in bats.

Cutaneous water loss and lipids of the stratum corneum in two syntopic species of bats

The lipid matrix of the stratum corneum (SC), the outer layer of the epidermis of mammals and birds, constitutes the barrier to diffusion of water vapor through the skin. The lipids of the SC are structured in the intercellular spaces of the mammalian epidermis in ordered layers, called lamellae, which have been postulated to prevent water loss. Lipids in the mammalian SC are mainly cholesterol, free fatty acids and ceramides, the latter forming the structural support for the lamellae. However, knowledge on how the lipid composition of the SC alters cutaneous water loss (CWL) in mammals is rudimentary, and is largely derived from studies on laboratory animals and humans. We measured CWL of individuals of two species of syntopic bats, Tadarida brasiliensis and Myotis velifer. In the first study of its kind on wild mammals, we correlated CWL with the lipid composition of the SC, measured using thin layer chromatography and high performance liquid chromatography coupled with atmospheric pressure photoionization mass spectrometry. Surface-specific CWL was 20.6% higher in M. velifer than in T. brasiliensis, although differences were not significant. Compared with individuals of M. velifer, individuals of T. brasiliensis had more classes, and a higher proportion, of polar ceramides in the SC, a feature associated with lower CWL. Individuals of T. brasiliensis also had a class of non-polar ceramides that presumably spans the lamellae and gives more cohesiveness to the lipid matrix of the SC. We conclude that qualitative and quantitative modifications of the lipid composition of the SC contribute to regulate CWL of these two species of bats.

Condition of Wings is an Important Criterion of Bat Health: A Response to Francl et al.

Wing damage is not used to confirm WNS: In response to elevated prevalence of wing damage in bats near WNS-affected hibernacula in 2008, we proposed a wing damage index (WDI) to objectively monitor wing conditions of bats in the field (Reichard and Kunz, 2009). Francl et al. suggested WDI is used ‘‘...as a proxy for measuring population-level infection’’ with WNS (Francl et al., 2011), but wing damage is only one field sign associated with WNS and does not confirm WNSpositive bats. Instead, WNS is diagnosed by histopathologic confirmation of cutaneous infections attributed to a recently introduced fungus, Geomyces destructans (Meteyer et al., 2009; USGS, 2011). Thus, we argue that suggesting use of WDI as a proxy for infection is inappropriate and misleading.

Vascular Contrast Enhanced Micro‐CT Imaging of “Radiators” in the Brazilian Free‐Tailed Bat (Tadarida brasiliensis)

The Brazilian free‐tailed bat (Tadarida brasiliensis) exhibits a highly vascularized, hairless thermal window (or “radiator”) on the proximal ventral surfaces of extended wings and body. We identified this character using thermal infrared imaging and investigated the vasculature using barium sulfate enhanced microcomputed tomography (micro‐CT). Micro‐CT images revealed unique arrangements of arteries and veins in the region of the radiator positioned perpendicular to the axis of the body. Coupling micro‐CT imaging with analysis of surface temperature profiles, we concluded that radiators aid in thermoregulation during flight in variable environments. This study represents the first application of contrast enhanced micro‐CT to visualize vasculature of bats and thus exhibits a promising technique for further investigations of cardiovascular function and anatomy in bats. Anat Rec, 2012.