Atlantic salmon Salmo salar do not prioritize digestion when energetic budgets are constrained by warming and hypoxia.

During summer, farmed Atlantic salmon (Salmo salar) can experience prolonged periods of warming and low aquatic oxygen levels due to climate change. This often results in a drop in feed intake; however, the physiological mechanism behind this behaviour is unclear. Digestion is a metabolically expensive process that can demand a high proportion of an animal's energy budget and might not be sustainable under future warming scenarios. We investigated the effects of elevated temperature and acute hypoxia on specific dynamic action (SDA; the energetic cost of digestion), and how much of the energy budget (i.e. aerobic scope, AS) was occupied by SDA in juvenile Atlantic salmon. AS was 9% lower in 21°C-acclimated fish compared to fish reared at their optimum temperature (15°C) and was reduced by ~50% by acute hypoxia (50% air saturation) at both temperatures. Furthermore, we observed an increase in peak oxygen uptake rate during digestion which occupied ~13% of the AS at 15°C and ~20% of AS at 21°C, and increased the total cost of digestion at 21°C. The minimum oxygen tolerance threshold in digesting fish was ~42% and ~53% at 15 and 21°C, respectively, and when digesting fish were exposed to acute hypoxia, gut transit was delayed. Thus, these stressors result in a greater proportion of the available energy budget being directed away from digestion. Moderate environmental hypoxia under both optimal and high temperatures severely impedes digestion and should be avoided to limit exacerbating temperature effects on fish growth.

Dramatic genome-wide reprogramming of mRNA in hypometabolic muscle.

In response to seasonal droughts, the green striped burrowing frog Cyclorana alboguttata enters a reversible hypometabolic state called aestivation where heart rate and oxygen consumption can be reduced despite warm (>25C°) ambient temperatures. With a view to understanding molecular mechanisms we profiled aestivating versus control gastrocnemius muscle using mRNA sequencing. This indicated an extensive metabolic reprogramming, with nearly a quarter of the entire transcriptome (3996 of 16,960 mRNA) exhibiting a nominal >2-fold change. Consistent with a physiological adaptation to spare carbohydrate reserves, carbohydrate catabolism was systemically downregulated. A 630-fold downregulation of ENO3 encoding the enolase enzyme was most striking. The 590 frog orthologs of mRNA encoding the mitoproteome were, viewed as a population, significantly downregulated during aestivation, although not to the same extent as mRNA encoding carbohydrate catabolism. Prominent examples include members of the TCA cycle (IDH2), electron transport chain (NDUFA6), the ATP synthase complex (ATP5F1B) and ADP/ATP intracellular transport (SLC25A4). Moreover, mRNA derived from the mt genome itself (e.g. mt-ND1) were also downregulated. Most prominent among the upregulated mRNA are those encoding aspects of regulated proteolysis including the proteosome (e.g. PSME4L), peptidases (USP25), atrogins (FBXO32) and ubiquitination (VCP). Finally, we note the ∼5-fold upregulation of the mRNA EIFG3 that encodes part of the EIF4F complex. This possesses global control of protein synthesis. Given protein synthesis is repressed in aestivating frogs this indicates the skeletal musculature is poised for accelerated translation of mRNA upon emergence, supporting a strategy to rapidly restore function when the summer rains come.

Cold-induced skin darkening does not protect amphibian larvae from UV-associated DNA damage.

Amphibian declines are sometimes correlated with increasing levels of ultraviolet radiation (UVR). While disease is often implicated in declines, environmental factors such as temperature and UVR play an important role in disease epidemiology. The mutagenic effects of UVR exposure on amphibians are worse at low temperatures. Amphibians from cold environments may be more susceptible to increasing UVR. However, larvae of some species demonstrate cold acclimation, reducing UV-induced DNA damage at low temperatures. Understanding of the mechanisms underpinning this response is lacking. We reared Limnodynastes peronii larvae in cool (15°C) or warm (25°C) waters before acutely exposing them to 1.5 h of high intensity (80 µW cm-2 ) UVBR. We measured the color of larvae and mRNA levels of a DNA repair enzyme. We reared larvae at 25°C in black or white containers to elicit a skin color response, and then measured DNA damage levels in the skin and remaining carcass following UVBR exposure. Cold-acclimated larvae were darker and displayed lower levels of DNA damage than warm-acclimated larvae. There was no difference in CPD-photolyase mRNA levels between cold- and warm-acclimated larvae. Skin darkening in larvae did not reduce their accumulation of DNA damage following UVR exposure. Our results showed that skin darkening does not explain cold-induced reductions in UV-associated DNA damage in L. peronii larvae. Beneficial cold-acclimation is more likely underpinned by increased CPD-photolyase abundance and/or increased photolyase activity at low temperatures.

Thermal compensation reduces DNA damage from UV radiation.

Increases in ultraviolet radiation (UVR) correlate spatially and temporally with global amphibian population declines and interact with other stressors such as disease and temperature. Declines have largely occurred in high-altitude areas associated with greater UVR and cooler temperatures. UVR is a powerful mutagenic harming organisms largely by damaging DNA. When acutely exposed to UVR at cool temperatures, amphibian larvae have increased levels of DNA damage. Amphibians may compensate for the depressive effects of temperature on DNA damage through acclimatisation, but it is unknown whether they have this capacity. We reared striped marsh frog larvae (Limnodynastes peronii) in warm (25 °C) and cool (15 °C) temperatures under a low or moderate daily dose of UVR (10 and 40 µW cm-2 UV-B for 1 h at midday, respectively) for 18-20 days and then measured DNA damage resulting from an acute high UVR dose (80 µW cm-2 UV-B for 1.5 h) at a range of temperatures (10, 15, 20, 25, and 30 °C). Larvae acclimated to 15 °C and exposed to UVR at 15 °C completely compensated UVR-induced DNA damage compared with 25 °C acclimated larvae exposed to UVR at 25 °C. Additionally, warm-acclimated larvae had higher DNA damage than cold-acclimated larvae across test temperatures, which indicated a cost of living in warmer temperatures. Larvae reared under elevated UVR levels showed no evidence of UVR acclimation resulting in lower DNA damage following high UVR exposure. Our finding that thermal acclimation in L. peronii larvae compensated UVR-induced DNA damage at low temperatures suggested that aquatic ectotherms living in cool temperatures may be more resilient to high UVR than previously realised. We suggested individuals or species with less capacity for thermal acclimation of DNA repair mechanisms may be more at risk if exposed to changing thermal and UVR exposure regimes.

Carryover effects from environmental change in early life: An overlooked driver of the amphibian extinction crisis?

Ecological carryover effects, or delayed effects of the environment on an organism's phenotype, are central predictors of individual fitness and a key issue in conservation biology. Climate change imposes increasingly variable environmental conditions that may be challenging to early life-history stages in animals with complex life histories, leading to detrimental physiological and fitness effects in later life. Yet, the latent nature of carryover effects, combined with the long temporal scales over which they can manifest, means that this phenomenon remains understudied and is often overlooked in short-term studies limited to single life-history stages. Herein, we review evidence for the physiological carryover effects induced by elevated ultraviolet radiation (UVR; 280-400 nm) as a potential contributor to recent amphibian population declines. UVR exposure causes a suite of molecular, cellular and physiological consequences known to underpin carryover effects in other taxa, but there is a lack of research linking embryonic and larval UVR exposures to fitness consequences post-metamorphosis in amphibians. We propose that the key impacts of UVR on disease-related amphibian declines are facilitated through carryover effects that bridge embryonic and larval UVR exposure with potential increased disease susceptibility post-metamorphosis. We conclude by identifying a practical direction for the study of ecological carryover effects in amphibians that could guide future ecological research in the broader field of conservation physiology. Only by addressing carryover effects can many of the mechanistic links between environmental change and population declines be elucidated.

Sublethal consequences of ultraviolet radiation exposure on vertebrates: Synthesis through meta-analysis.

Ultraviolet radiation (UVR) from the sun is a natural daytime stressor for vertebrates in both terrestrial and aquatic ecosystems. UVR effects on the physiology of vertebrates manifest at the cellular level, but have bottom-up effects at the tissue level and on whole-animal performance and behaviours. Climate change and habitat loss (i.e. loss of shelter from UVR) could interact with and exacerbate the genotoxic and cytotoxic impacts of UVR on vertebrates. Therefore, it is important to understand the range and magnitude of effects that UVR can have on a diversity of physiological metrics, and how these may be shaped by taxa, life stage or geographical range in the major vertebrate groups. Using a meta-analytical approach, we used 895 observations from 47 different vertebrate species (fish, amphibian, reptile and bird), and 51 physiological metrics (i.e. cellular, tissue and whole-animal metrics), across 73 independent studies, to elucidate the general patterns of UVR effects on vertebrate physiology. We found that while UVR's impacts on vertebrates are generally negative, fish and amphibians were the most susceptible taxa, adult and larvae were the most susceptible life stages, and animals inhabiting temperate and tropical latitudes were the most susceptible to UVR stress. This information is critical to further our understanding of the adaptive capacity of vulnerable taxon to UVR stress, and the wide-spread sublethal physiological effects of UVR on vertebrates, such as DNA damage and cellular stress, which may translate up to impaired growth and locomotor performance. These impairments to individual fitness highlighted by our study may potentially cause disruptions at the ecosystem scale, especially if the effects of this pervasive diurnal stressor are exacerbated by climate change and reduced refuge due to habitat loss and degradation. Therefore, conservation of habitats that provide refuge to UVR stress will be critical to mitigate stress from this pervasive daytime stressor.

Biological mechanisms matter in contemporary wildlife conservation.

Given limited resources for wildlife conservation paired with an urgency to halt declines and rebuild populations, it is imperative that management actions are tactical and effective. Mechanisms are about how a system works and can inform threat identification and mitigation such that conservation actions that work can be identified. Here, we call for a more mechanistic approach to wildlife conservation and management where behavioral and physiological tools and knowledge are used to characterize drivers of decline, identify environmental thresholds, reveal strategies that would restore populations, and prioritize conservation actions. With a growing toolbox for doing mechanistic conservation research as well as a suite of decision-support tools (e.g., mechanistic models), the time is now to fully embrace the concept that mechanisms matter in conservation ensuring that management actions are tactical and focus on actions that have the potential to directly benefit and restore wildlife populations.

Body size dictates physiological and behavioural responses to hypoxia and elevated water temperatures in Murray cod (Maccullochella peelii).

Increasing drought frequency and duration pose a significant threat to fish species in dryland river systems. As ectotherms, fish thermal and hypoxia tolerances directly determine the capacity of species to persist in these environments during low flow periods when water temperatures are high and waterbodies become highly stratified. Chronic thermal stress can compound the impacts of acute hypoxic events on fish resulting in significant fish mortality; however, it is not known if all size classes are equally susceptible, or if the allometric scaling of physiological processes means some size classes are disproportionately affected. We investigated the physiological responses of Murray cod (Maccullochella peelii) over a four-fold body size range (0.2-3000 g) to acute changes in water temperature and oxygen concentration following 4 weeks of acclimation to representative spring (20°C) and summer (28°C) water temperatures. We recorded maximum thermal tolerance (CT max), oxygen limited thermal tolerance (PCTmax ), lowest tolerable oxygen level (as the oxygen level at which lose equilibrium; O2,LOE), gill ventilation rates and aerial surface respiration threshold, blood oxygen transport capacity and lactate accumulation. Acclimation to elevated water temperatures improved thermal and hypoxia tolerance metrics across all size classes. However, body size significantly affected thermal and hypoxia responses. Small M. peelii were significantly less hypoxia tolerant than larger individuals, while larger fish were significantly less thermal tolerant than smaller fish. Hypoxia constrained thermal tolerance in M. peelii, with both small and large fish disproportionally compromised relative to mid-sized fish. Our findings indicate that both very small/young (larvae, fry, fingerlings) and very large/older M. peelii in dryland rivers are at significant risk from the combined impacts of a warming and drying climate and water extraction. These data will inform policy decisions that serve to balance competing demands on precious freshwater resources.

Temperature causes species-specific responses to UV-induced DNA damage in amphibian larvae.

Anthropogenic ozone depletion has led to a 2-5% increase in ultraviolet B radiation (UVBR) levels reaching the earth's surface. Exposure to UVBR causes harmful DNA damage in amphibians, but this is minimized by DNA repair enzymes such as thermally sensitive cyclobutane pyrimidine dimer (CPD)-photolyase, with cool temperatures slowing repair rates. It is unknown whether amphibian species differ in the repair response to a given dose of UVBR across temperatures. We reared larvae of three species (Limnodynastes peronii, Limnodynastes tasmaniensis and Platyplectrum ornatum) at 25°C and acutely exposed them to 80 µW cm-2 UVBR for 2 h at either 20°C or 30°C. UVBR-mediated DNA damage was measured as larvae repaired damage in photoreactive light at their exposure temperatures. Cool temperatures increased DNA damage in two species and slowed DNA repair rate in P. ornatum. The magnitude of DNA damage incurred from UVBR was species-specific. Platyplectrum ornatum had the lowest CPDs and DNA repair rates, and the depressive effects of low temperature on photorepair were greater in L. tasmaniensis. Considering the susceptibility of most aquatic organisms to UVBR, this research highlighted a need to consider the complexity of species-specific physiology when forecasting the influence of changing UVBR and temperature in aquatic ecosystems.

Early exposure to UV radiation causes telomere shortening and poorer condition later in life.

Determining the contribution of elevated ultraviolet-B radiation (UVBR; 280-315 nm) to amphibian population declines is being hindered by a lack of knowledge about how different acute UVBR exposure regimes during early life-history stages might affect post-metamorphic stages via long-term carryover effects. We acutely exposed tadpoles of the Australian green tree frog (Litoria caerulea) to a combination of different UVBR irradiances and doses in a multi-factorial laboratory experiment, and then reared them to metamorphosis in the absence of UVBR to assess carryover effects in subsequent juvenile frogs. Dose and irradiance of acute UVBR exposure influenced carryover effects into metamorphosis in somewhat opposing manners. Higher doses of UVBR exposure in larvae yielded improved rates of metamorphosis. However, exposure at a high irradiance resulted in frogs metamorphosing smaller in size and in poorer condition than frogs exposed to low and medium irradiance UVBR as larvae. We also demonstrate some of the first empirical evidence of UVBR-induced telomere shortening in vivo, which is one possible mechanism for life-history trade-offs impacting condition post-metamorphosis. These findings contribute to our understanding of how acute UVBR exposure regimes in early life affect later life-history stages, which has implications for how this stressor may shape population dynamics.

UV exposure causes energy trade-offs leading to increased chytrid fungus susceptibility in green tree frog larvae.

Levels of ultraviolet (UV) radiation have increased in many parts of the world due to the anthropogenic destruction of the ozone layer. UV radiation is a potent immunosuppressant and can increase the susceptibility of animal hosts to pathogens. UV radiation can directly alter immune function via immunosuppression and photoimmunotolerance; however, UV may also influence pathogen defences by affecting the distribution of energy resources among competing physiological processes. Both defence against UV damage and repair of incurred damage, as well as the maintenance of immune defences and responding to an immune challenge, are energetically expensive. These competing demands for finite energy resources could trade off against one another, resulting in sub-optimal performance in one or both processes. We examined the potential for a disease-related energy trade-off in green tree frog (Litoria caerulea) larvae. Larvae were reared under high- or low-UV conditions for 12 weeks during which time we measured growth rates, metabolic rate and susceptibility to the amphibian fungal pathogen, Batrachochytrium dendrobatidis (Bd). We found that larvae exposed to high levels of UV radiation had higher rates of energy expenditure than those exposed to low UV levels; however, UV exposure did not affect growth rates or developmental timings. Larvae exposed to high UV radiation also experienced greater Bd infection rates and carried a higher infection burden than those not exposed to elevated UV radiation. We propose that the increased energetic costs of responding to UV radiation were traded off against immune defences to protect larval growth rates. These findings have important implications for the aetiology of some Bd-associated amphibian declines, particularly in montane environments where Bd infections are most severe and where UV levels are highest.

The role of environmental calcium in the extreme acid tolerance of northern banjo frog (Limnodynastes terraereginae) larvae.

Many aquatically respiring animals acutely exposed to low pH waters suffer inhibition of ion uptake and loss of branchial (gill) epithelial integrity, culminating in a fatal loss of body Na+. Environmental calcium levels ([Ca2+]e) are pivotal in maintaining branchial junction integrity, with supplemental Ca2+ reversing the negative effects of low pH in some animals. Tolerance of some naturally acidic environments by aquatic animals is further complicated by low [Ca2+]e, yet many of these environments are surprisingly biodiverse. How animals overcome the damaging actions of low pH and low environmental Ca2+ remains unknown. We examined the effects of [Ca2+]e on the response to low pH in larvae of the highly acid-tolerant frog Limnodynastes terraereginae. Acute exposure to low pH water in the presence of low (5 µmol l-1) [Ca2+]e increased net Na+ efflux. Provision of additional [Ca2+]e reduced net Na+ efflux, but the effect was saturable. Acclimation to both low and high (250 µmol l-1) [Ca2+]e improved the resistance of larvae to Na+ efflux at low pH. Exposure to the Ca2+ channel inhibitor ruthenium red resulted in an abrupt loss of tolerance in low pH-acclimated larvae. Acclimation to acidic water increased branchial gene expression of the intracellular Ca2+ transport protein calbindin, consistent with a role for increased transcellular Ca2+ trafficking in the tolerance of acidic water. This study supports a role for [Ca2+]e in promoting branchial integrity and highlights a potential mechanism via the maintenance of transcellular Ca2+ uptake in the acid tolerance of L. terraereginae larvae.

Fire and rain: A systematic review of the impacts of wildfire and associated runoff on aquatic fauna.

Climate and land-use changes are expected to increase the future occurrence of wildfires, with potentially devastating consequences for freshwater species and ecosystems. Wildfires that burn in close proximity to freshwater systems can significantly alter the physicochemical properties of water. Following wildfires and heavy rain, freshwater species must contend with complex combinations of wildfire ash components (nutrients, polycyclic aromatic hydrocarbons, and metals), altered light and thermal regimes, and periods of low oxygen that together can lead to mass mortality events. However, the responses of aquatic fauna to wildfire disturbances are poorly understood. Here we provide a systematic review of available evidence on how aquatic animals respond to and recover from wildfire disturbance. Two databases (Web of Science and Scopus) were used to identify key literature. A total of 83 studies from across 11 countries were identified to have assessed the risk of wildfires on aquatic animals. We provide a summary of the main ecosystem-level changes associated with wildfires and the main responses of aquatic fauna to such disturbances. We pay special focus to physiological tools and biomarkers used to assess how wildfires impact aquatic animals. We conclude by providing an overview of how physiological biomarkers can further our understanding of wildfire-related impacts on aquatic fauna, and how different physiological tools can be incorporated into management and conservation plans and serve as early warning signs of wildfire disturbances.

Se espera que el cambio climático y el cambio en el uso de suelo aumentaran la ocurrencia de incendios forestales, con consecuencias potencialmente devastadoras para las especies de agua dulce y los ecosistemas. Los incendios forestales que arden cerca de los sistemas de agua dulce pueden alterar significativamente las propiedades fisicoquímicas del agua. Después de los incendios forestales y llueves fuertes, las especies de agua dulce lidian con combinaciones complejas de componentes de cenizas de incendios forestales (nutrientes, sedimentos, hidrocarburos aromáticos policíclicos y metales), regímenes de luz y térmicos alterados y períodos de bajo oxígeno que, en conjunto, pueden conducir a eventos de mortalidad masiva. Sin embargo, las respuestas de la fauna acuática a las perturbaciones de los incendios forestales son poco conocidas. Aquí proporcionamos una revisión sistemática de la evidencia disponible sobre cómo los animales acuáticos responden y se recuperan de la perturbación de los incendios forestales. Se utilizaron dos bases de datos (Web of Science y Scopus) para identificar la literatura clave. Se identificaron un total de 83 estudios de 11 países que habían evaluado el riesgo de incendios forestales en animales acuáticos. Proporcionamos un resumen de los principales cambios a nivel de ecosistema asociados con los incendios forestales y las principales respuestas de la fauna acuática a tales perturbaciones. Prestamos especial atención a las herramientas fisiológicas y los biomarcadores que se utilizan para evaluar cómo los incendios forestales afectan a los animales acuáticos. Concluimos proporcionando una descripción general de cómo los biomarcadores fisiológicos pueden mejorar nuestra comprensión de los impactos relacionados con los incendios forestales en la fauna acuática, y cómo se pueden incorporar diferentes herramientas fisiológicas en los planes de gestión y conservación y servir como señales de alerta temprana de las perturbaciones de los incendios forestales.

Can slowing the rate of water temperature decline be utilized to reduce the impacts of cold water pollution from dam releases on fish physiology and performance?

Cold water pollution (CWP) is caused by releases of unseasonably cold water from large, thermally stratified dams. Rapid and prolonged decreases in water temperature can have depressive effects on the metabolism, growth and swimming performance of fish. However, it is unknown if reducing the rate of temperature decrease could mitigate these negative effects by allowing thermal acclimation/acclimatization to occur. This study investigated the rate of temperature decrease as a potential CWP mitigation strategy in juvenile Murray cod Maccullochella peelii. M. peelii were exposed to a gradual, intermediate or rapid temperature decrease from 24 to 14°C. Energetic costs, locomotor performance, growth and survival were measured to determine if the initial thermal regime affected the thermal acclimation capacity of M. peelii. Cold exposure had significant acute and lasting depressive effects regardless of the rate of temperature decrease, although M. peelii showed varying degrees of thermal compensation in swimming performance and metabolism after 8 weeks of exposure to low temperatures. The short-term effects of CWP-like reductions in temperature are significant, but over time M. peelii can offset some of the depressive effects of CWP through thermal plasticity. This study highlights the importance of understanding physiological responses of fish to inform management and conservation. We conclude that rate of water temperature decline cannot be used to mitigate the sublethal effects of CWP on juvenile M. peelii but may still be useful for managing the negative effects in other native Australian fish species.

Applied ecoimmunology: using immunological tools to improve conservation efforts in a changing world.

Ecoimmunology is a rapidly developing field that explores how the environment shapes immune function, which in turn influences host-parasite relationships and disease outcomes. Host immune defence is a key fitness determinant because it underlies the capacity of animals to resist or tolerate potential infections. Importantly, immune function can be suppressed, depressed, reconfigured or stimulated by exposure to rapidly changing environmental drivers like temperature, pollutants and food availability. Thus, hosts may experience trade-offs resulting from altered investment in immune function under environmental stressors. As such, approaches in ecoimmunology can provide powerful tools to assist in the conservation of wildlife. Here, we provide case studies that explore the diverse ways that ecoimmunology can inform and advance conservation efforts, from understanding how Galapagos finches will fare with introduced parasites, to using methods from human oncology to design vaccines against a transmissible cancer in Tasmanian devils. In addition, we discuss the future of ecoimmunology and present 10 questions that can help guide this emerging field to better inform conservation decisions and biodiversity protection. From better linking changes in immune function to disease outcomes under different environmental conditions, to understanding how individual variation contributes to disease dynamics in wild populations, there is immense potential for ecoimmunology to inform the conservation of imperilled hosts in the face of new and re-emerging pathogens, in addition to improving the detection and management of emerging potential zoonoses.

Ultraviolet-B irradiance and cumulative dose combine to determine performance and survival.

Despite decades of research, the role of elevated solar ultraviolet-B radiation (UVBR; 280-315 nm) in shaping amphibian populations remains ambiguous. These difficulties stem partly from a poor understanding of which parameters of UVBR exposure - dose, irradiance, and time interval - determine UVBR exposure health risk, and the potentially erroneous assumption that effects are proportional to the dose of exposure, irrespective of the administered regime (Bunsen-Roscoe Law of Reciprocity; BRL). We tested if the BRL holds with respect to UVBR-induced physiological effects in amphibians by acutely exposing tadpoles of the Australian green tree frog (Litoria caerulea) to a combination of different UVBR irradiances and doses in a fully factorial experiment. The BRL was invalid across all metrics assessed, with UVBR irradiance influencing the effects of a given dose on growth, coloration and burst swimming performance of larvae. We demonstrated some of the first empirical evidence for beneficial physiological effects of UVBR exposure in a larval amphibian, with improvements in growth, burst swimming performance and survival at the highest UVBR doses, contrary to hypotheses. Our findings demonstrate the species-specific nature of amphibian responses to UVBR, and the importance of UVBR irradiance in influencing the long-term physiological effects of a given dose of radiation. This work enhances our understanding of which parameters of complex UVBR exposures determine amphibian health risk.

One hundred research questions in conservation physiology for generating actionable evidence to inform conservation policy and practice.

Environmental change and biodiversity loss are but two of the complex challenges facing conservation practitioners and policy makers. Relevant and robust scientific knowledge is critical for providing decision-makers with the actionable evidence needed to inform conservation decisions. In the Anthropocene, science that leads to meaningful improvements in biodiversity conservation, restoration and management is desperately needed. Conservation Physiology has emerged as a discipline that is well-positioned to identify the mechanisms underpinning population declines, predict responses to environmental change and test different in situ and ex situ conservation interventions for diverse taxa and ecosystems. Here we present a consensus list of 10 priority research themes. Within each theme we identify specific research questions (100 in total), answers to which will address conservation problems and should improve the management of biological resources. The themes frame a set of research questions related to the following: (i) adaptation and phenotypic plasticity; (ii) human-induced environmental change; (iii) human-wildlife interactions; (iv) invasive species; (v) methods, biomarkers and monitoring; (vi) policy, engagement and communication; (vii) pollution; (viii) restoration actions; (ix) threatened species; and (x) urban systems. The themes and questions will hopefully guide and inspire researchers while also helping to demonstrate to practitioners and policy makers the many ways in which physiology can help to support their decisions.

Exposure to Nitrate Increases Susceptibility to Hypoxia in Fish.

AbstractAquatic hypoxic events are increasing in frequency and intensity as concentrations of nutrients, such as nitrate, continue to rise from human activities. Many fish species can alter their behavior and physiology to cope with drops in oxygen, but these compensatory strategies may be compromised under high levels of nitrate pollution. Hence, we investigated whether exposure to elevated nitrate concentrations affects key behavioral (avoidance and aquatic surface respiration [ASR]) and physiological (hemoglobin and hematocrit levels, ventilation frequency, and burst and prolonged swimming performance) responses of fish to mitigate the impacts of acute hypoxia. Juvenile silver perch (Bidyanus bidyanus) were exposed to one of three nitrate concentrations (0, 50, or 100 mg NO3- L-1) for 3 wk, after which behavioral and physiological responses of fish to progressive hypoxia were assessed. Fish exposed to nitrate utilized ASR at a higher threshold of partial pressure of oxygen during progressive hypoxia compared with control animals but did not alter behavioral avoidance of low oxygen levels. In these nitrate-exposed fish, the early onset of ASR behaviors is likely a behavioral, compensatory strategy to cope with nitrate-induced physiological disruptions, namely, increases in ventilation frequency and lower levels of hemoglobin and hematocrit. The physiological constraints placed by nitrate and acute hypoxia exposures manifested to lower the swimming performance of silver perch. Collectively, these data suggest that exposure to elevated nitrate is likely to disrupt key behavioral and physiological strategies used by fish to cope with short-term hypoxia.

Conservation physiology and the COVID-19 pandemic.

The COVID-19 pandemic and associated public health measures have had unanticipated effects on ecosystems and biodiversity. Conservation physiology and its mechanistic underpinnings are well positioned to generate robust data to inform the extent to which the Anthropause has benefited biodiversity through alterations in disturbance-, pollution- and climate change-related emissions. The conservation physiology toolbox includes sensitive biomarkers and tools that can be used both retroactively (e.g. to reconstruct stress in wildlife before, during and after lockdown measures) and proactively (e.g. future viral waves) to understand the physiological consequences of the pandemic. The pandemic has also created new risks to ecosystems and biodiversity through extensive use of various antimicrobial products (e.g. hand cleansers, sprays) and plastic medical waste. Conservation physiology can be used to identify regulatory thresholds for those products. Moreover, given that COVID-19 is zoonotic, there is also opportunity for conservation physiologists to work closely with experts in conservation medicine and human health on strategies that will reduce the likelihood of future pandemics (e.g. what conditions enable disease development and pathogen transfer) while embracing the One Health concept. The conservation physiology community has also been impacted directly by COVID-19 with interruptions in research, training and networking (e.g. conferences). Because this is a nascent discipline, it will be particularly important to support early career researchers and ensure that there are recruitment pathways for the next generation of conservation physiologists while creating a diverse and inclusive community. We remain hopeful for the future and in particular the ability of the conservation physiology community to deliver relevant, solutions-oriented science to guide decision makers particularly during the important post-COVID transition and economic recovery.

Physiological and morphological correlates of extreme acid tolerance in larvae of the acidophilic amphibian Litoria cooloolensis.

The Cooloola sedgefrog (Litoria cooloolensis) is one of a number of frog species endemic to the coastal sandy lowlands of east Australia exhibiting remarkable tolerance to dilute waters of low pH (< pH 3.5). To investigate the physiological and morphological underpinnings of acid tolerance in L. cooloolensis larvae, we compared Na+ balance, uptake and efflux rates, and gill and skin morphology in larvae reared in circum-neutral (pH 6.5) and pH 3.5 water. We hypothesised that L. cooloolensis larvae would be more resistant to ionregulatory disturbance and epithelial damage at low pH relative to acid-sensitive species. Net Na+ flux rates were not significantly different from zero in L. cooloolensis larvae reared at pH 3.5 and in acid-naïve animals maintained in pH 6.5 water. Animals reared at pH 6.5 and acutely exposed to pH 3.5, however, exhibited a net loss of Na+ due to inhibition of Na+ uptake. In contrast, L. cooloolensis larvae reared at pH 3.5 maintained Na+ balance at pH 3.5 and did not exhibit inhibition of Na+ uptake at this pH. Investigation of Na+ transport kinetics and the morphology of the gills and integument suggests tolerance of L. cooloolensis larvae to low pH may be attributed to a high capacity for branchial Na+ uptake, increased tight junction length and elevated mucus production at the gills and integument. These factors confer resistance to acid damage and disruption of ionic homeostasis which would otherwise result in the death of amphibian larvae exposed to waters of pH 4.0 and less.