Glutamatergic Receptors Modulate Normoxic but Not Hypoxic Ventilation and Metabolism in Naked Mole Rats

Naked mole rats (Heterocephalus glaber) are among the most hypoxia-tolerant mammals, but their physiological responses to acute and chronic sustained hypoxia (CSH), and the molecular underpinnings of these responses, are poorly understood. In the present study we evaluated the acute hypoxic ventilatory response and the occurrence of ventilatory acclimatization to hypoxia following CSH exposure (8–10 days in 8% O2) of naked mole rats. We also investigated the role of excitatory glutamatergic signaling in the control of ventilation and metabolism in these conditions. Animals acclimated to normoxia (control) or CSH and then exposed to acute hypoxia (7% O2 for 1 h) exhibited elevated tidal volume (VT), but decreased breathing frequency (fR). As a result, total ventilation (V.E) remained unchanged. Conversely, VT was lower in CSH animals relative to controls, suggesting that there is ventilatory plasticity following acclimatization to chronic hypoxia. Both control and CSH-acclimated naked mole rats exhibited similar 60–65% decreases in O2 consumption rate during acute hypoxia, and as a result their air convection requirement (ACR) increased ∼2.4 to 3-fold. Glutamatergic receptor inhibition decreased fR, V.E, and the rate of O2 consumption in normoxia but did not alter these ventilatory or metabolic responses to acute hypoxia in either the control or CSH groups. Taken together, these findings indicate that ventilatory acclimatization to hypoxia is atypical in naked mole rats, and glutamatergic signaling is not involved in their hypoxic ventilatory or metabolic responses to acute or chronic hypoxia

Similar hibernation physiology in bats across broad geographic ranges

This is a post-peer-review, pre-copyedit version of an article published in Journal of Comparative Physiology B. The final authenticated version is available online at: http://dx.doi.org/10.1007/s00360-021-01400-xSpecies 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.Department of Defense Strategic Environmental Research and Development Program || United States Fish and Wildlife Service, Grant F17AP00593 || Texas Tech University || Alberta Conservation Association

Data from: Adenosine receptors mediate the hypoxic ventilatory response but not the hypoxic metabolic response in the naked mole rat during acute hypoxia

Naked mole rats are the most hypoxia-tolerant mammals identified; however, the mechanisms underlying this tolerance are poorly understood. Using whole-animal plethysmography and open-flow respirometry, we examined the hypoxic metabolic response (HMR), hypoxic ventilatory response (HVR) and hypoxic thermal response in awake, freely behaving naked mole rats exposed to 7% O2 for 1 h. Metabolic rate and ventilation each reversibly decreased 70% in hypoxia (from 39.6 ± 2.9 to 12.1 ± 0.3 ml O2 min−1 kg−1, and 1412 ± 244 to 417 ± 62 ml min−1 kg−1, respectively; p < 0.05), whereas body temperature was unchanged and animals remained awake and active. Subcutaneous injection of the general adenosine receptor antagonist aminophylline (AMP; 100 mg kg−1, in saline), but not control saline injections, prevented the HVR but had no effect on the HMR. As a result, AMP-treated naked mole rats exhibited extreme hyperventilation in hypoxia. These animals were also less tolerant to hypoxia, and in some cases hypoxia was lethal following AMP injection. We conclude that in naked mole rats (i) hypoxia tolerance is partially dependent on profound hypoxic metabolic and ventilatory responses, which are equal in magnitude but occur independently of thermal changes in hypoxia, and (ii) adenosine receptors mediate the HVR but not the HMR

Bats increased foraging activity at experimental prey patches near hibernacula

Abstract: Emerging infectious diseases in wildlife can threaten vulnerable host populations. Actions targeting habitat improvements to aid population resilience and recovery may be beneficial long‐term strategies, yet testing the efficacy of such strategies before major conservation investments are made can be challenging. The disease white‐nose syndrome (WNS) has caused severe declines in several species of North American hibernating bats. We tested a novel conservation approach targeted at improving foraging conditions near bat hibernacula by experimentally manipulating insect density in the pre‐hibernation fattening period and spring emergence recovery period. We measured foraging (feeding buzzes) and echolocation activity of little brown bats Myotis lucifugus at ultraviolet (UV) light lures to determine behavioural response to augmented foraging conditions and characterized insect availability at UV light lures. In the fall, bat foraging activity was three times greater (95% CI: 1.5–5.8; p&nbsp;=&nbsp;0.002) when UV lights were on, but there was no statistical support for differences in echolocation activity response when our experimental design alternated between nights with lights on and off. In the spring, we allowed UV light lures to run consistently each night and compared with a control location in similar habitat. Bat foraging activity was 8.5 times greater (95% CI: 4.5–16.0; p &lt; 0.0001) and echolocation activity was 4.4 times higher (95% CI: 3.0–6.5; p &lt; 0.0001) at UV light lures in the spring experiment. In both the fall and spring, UV light lures resulted in concentrated insect availability, attracting primarily moths (Order: Lepidoptera). In both seasons, nightly temperature had a strong influence on bat foraging, echolocation and insect activity. We show that a bat species threatened by WNS used enhanced foraging habitats near hibernacula during the critical pre‐ and post‐hibernation phases of their annual cycle. While light lures are unlikely to be a long‐term management strategy, our experiment provides initial evidence that bats behaviourally respond with increased foraging activity in areas with augmented insect prey availability. Our experimental results support developing management strategies focused on habitat protection, including restoration and enhancement of foraging habitats, in the immediate vicinity of bat hibernacula

Bats increased foraging activity at experimental prey patches near hibernacula

Emerging infectious diseases in wildlife can threaten vulnerable host populations. Actions targeting habitat improvements to aid population resilience and recovery may be beneficial long-term strategies, yet testing the efficacy of such strategies before major conservation investments are made can be challenging. The disease white-nose syndrome (WNS) has caused severe declines in several species of North American hibernating bats. We tested a novel conservation approach targeted at improving foraging conditions near bat hibernacula by experimentally manipulating insect density in the pre-hibernation fattening period and spring emergence recovery period. We measured foraging (feeding buzzes) and echolocation activity of little brown bats Myotis lucifugus at ultraviolet (UV) light lures to determine behavioural response to augmented foraging conditions and characterized insect availability at UV light lures. In the fall, bat foraging activity was three times greater (95% CI: 1.5–5.8; p = 0.002) when UV lights were on, but there was no statistical support for differences in echolocation activity response when our experimental design alternated between nights with lights on and off. In the spring, we allowed UV light lures to run consistently each night and compared with a control location in similar habitat. Bat foraging activity was 8.5 times greater (95% CI: 4.5–16.0; p \u3c 0.0001) and echolocation activity was 4.4 times higher (95% CI: 3.0–6.5; p \u3c 0.0001) at UV light lures in the spring experiment. In both the fall and spring, UV light lures resulted in concentrated insect availability, attracting primarily moths (Order: Lepidoptera). In both seasons, nightly temperature had a strong influence on bat foraging, echolocation and insect activity. We show that a bat species threatened by WNS used enhanced foraging habitats near hibernacula during the critical pre-and post-hibernation phases of their annual cycle. While light lures are unlikely to be a long-term management strategy, our experiment provides initial evidence that bats behaviourally respond with increased foraging activity in areas with augmented insect prey availability. Our experimental results support developing management strategies focused on habitat protection, including restoration and enhancement of foraging habitats, in the immediate vicinity of bat hibernacula

White-Nose Syndrome Disrupts the Splenic Lipidome of Little Brown Bats (Myotis lucifugus) at Early Disease Stages

White-nose syndrome (WNS)-positive little brown bats (Myotis lucifugus) may exhibit immune responses including increased cytokine and pro-inflammatory mediator gene levels. Bioactive lipid mediators (oxylipins) formed by enzymatic oxidation of polyunsaturated fatty acids can contribute to these immune responses, but have not been investigated in WNS pathophysiology. Nonenzymatic conversion of polyunsaturated fatty acids can also occur due to reactive oxygen species, however, these enantiomeric isomers will lack the same signaling properties. In this study, we performed a series of targeted lipidomic approaches on laboratory Pseudogymnoascus destructans-inoculated bats to assess changes in their splenic lipidome, including the formation of lipid mediators at early stages of WNS. Hepatic lipids previously identified were also resolved to a higher structural detail. We compared WNS-susceptible M. lucifugus to a WNS-resistant species, the big brown bat (Eptesicus fuscus). Altered splenic lipid levels were only observed in M. lucifugus. Differences in splenic free fatty acids included both omega-3 and omega-6 compounds. Increased levels of an enantiomeric monohydroxy DHA mixture were found, suggesting nonenzymatic formation. Changes in previously identified hepatic lipids were confined to omega-3 constituents. Together, these results suggest that increased oxidative stress, but not an inflammatory response, is occurring in bats at early stages of WNS that precedes fat depletion. These data have been submitted to metabolomics workbench and assigned a study number ST002304

Adipose Tissues from Human and Bat-Derived Cell Lines Support Ebola Virus Infection

Ebola virus is a zoonotic pathogen with a geographic range covering diverse ecosystems that are home to many potential reservoir species. Although researchers have detected Ebola virus RNA and serological evidence of previous infection in different rodents and bats, the infectious virus has not been isolated. The field is missing critical knowledge about where the virus is maintained between outbreaks, either because the virus is rarely encountered, overlooked during sampling, and/or requires specific unknown conditions that regulate viral expression. This study assessed adipose tissue as a previously overlooked tissue capable of supporting Ebola virus infection. Adipose tissue is a dynamic endocrine organ helping to regulate and coordinate homeostasis, energy metabolism, and neuroendocrine and immune functions. Through in vitro infection of human and bat (Eptesicus fuscus) brown adipose tissue cultures using wild-type Ebola virus, this study showed high levels of viral replication for 28 days with no qualitative indicators of cytopathic effects. In addition, alterations in adipocyte metabolism following long-term infection were qualitatively observed through an increase in lipid droplet number while decreasing in size, a harbinger of lipolysis or adipocyte browning. The finding that bat and human adipocytes are susceptible to Ebola virus infection has important implications for potential tissue tropisms that have not yet been investigated. Additionally, the findings suggest how the metabolism of this tissue may play a role in pathogenesis, viral transmission, and/or zoonotic spillover events

White-Nose Syndrome Disrupts the Splenic Lipidome of Little Brown Bats (Myotis lucifugus) at Early Disease Stages

White-nose syndrome (WNS)-positive little brown bats (Myotis lucifugus) may exhibit immune responses including increased cytokine and pro-inflammatory mediator gene levels. Bioactive lipid mediators (oxylipins) formed by enzymatic oxidation of polyunsaturated fatty acids can contribute to these immune responses, but have not been investigated in WNS pathophysiology. Nonenzymatic conversion of polyunsaturated fatty acids can also occur due to reactive oxygen species, however, these enantiomeric isomers will lack the same signaling properties. In this study, we performed a series of targeted lipidomic approaches on laboratory Pseudogymnoascus destructans-inoculated bats to assess changes in their splenic lipidome, including the formation of lipid mediators at early stages of WNS. Hepatic lipids previously identified were also resolved to a higher structural detail. We compared WNS-susceptible M. lucifugus to a WNS-resistant species, the big brown bat (Eptesicus fuscus). Altered splenic lipid levels were only observed in M. lucifugus. Differences in splenic free fatty acids included both omega-3 and omega-6 compounds. Increased levels of an enantiomeric monohydroxy DHA mixture were found, suggesting nonenzymatic formation. Changes in previously identified hepatic lipids were confined to omega-3 constituents. Together, these results suggest that increased oxidative stress, but not an inflammatory response, is occurring in bats at early stages of WNS that precedes fat depletion. These data have been submitted to metabolomics workbench and assigned a study number ST002304

White-nose syndrome is associated with increased replication of a naturally persisting coronaviruses in bats

Spillover of viruses from bats to other animals may be associated with increased contact between them, as well as increased shedding of viruses by bats. Here, we tested the prediction that little brown bats (Myotis lucifugus) co-infected with the M. lucifugus coronavirus (Myl-CoV) and with Pseudogymnoascus destructans (Pd), the fungus that causes bat white-nose syndrome (WNS), exhibit different disease severity, viral shedding and molecular responses than bats infected with only Myl-CoV or only P. destructans. We took advantage of the natural persistence of Myl-CoV in bats that were experimentally inoculated with P. destructans in a previous study. Here, we show that the intestines of virus-infected bats that were also infected with fungus contained on average 60-fold more viral RNA than bats with virus alone. Increased viral RNA in the intestines correlated with the severity of fungus-related pathology. Additionally, the intestines of bats infected with fungus exhibited different expression of mitogen-activated protein kinase pathway and cytokine related transcripts, irrespective of viral presence. Levels of coronavirus antibodies were also higher in fungal-infected bats. Our results suggest that the systemic effects of WNS may down-regulate anti-viral responses in bats persistently infected with M. lucifugus coronavirus and increase the potential of virus shedding