Upper thermal limits are 'hard-wired' across body mass but not populations of an estuarine fish.

Climate warming is seeing temperatures breach exceptional thresholds as the frequency and intensity of heat waves increase. Efforts to forecast species vulnerability to climate warming often focus on upper thermal limits threatening survival, overlooking the role of intraspecific variation in determining vulnerability. Using an estuarine fish (black bream, Acanthopagrus butcheri) as a model, we explore how intraspecific variation in body mass and among populations affects upper thermal tolerance. Upper thermal limits were quantified using critical thermal maxima (CTmax) of wild fish. We used a ∼500 g (mean = 52.4 g, range = 0.57-541 g) mass range to test the relationship between body mass and thermal tolerance. Four distinct black bream populations were chosen along a 5° latitudinal cline to explore population differences in thermal limits. Contrary to expectations, there was no effect of body mass on upper thermal limits. However, significant population differences in thermal tolerance were observed that correlate with mean habitat temperatures. Specifically, the southern population had a significantly lower CTmax (35.57 ± 0.43 °C) compared to northern (36.32 ± 0.70 °C) and mid-latitude (36.36 ± 1.15 °C) populations. These data underscore the importance of observing intraspecific variation in thermal limits to reveal the capabilities of individuals within a species to cope with climate warming and improve the management of at-risk life stages and populations.

Deteriorating waterways: The effect of nitrate pollution on the development and physiology of the endangered southern bell frog (Litoria raniformis).

Nitrogen-based fertilizers can increase agricultural yields and crop quality, but this comes at the risk of contaminating nearby waterways. Nitrate is the most stable and abundant form of inorganic nitrogen in the environment and chronic exposure can impair performance and fitness in aquatically respiring species. But it remains unknown if these impairments are linked to disruptions in energy homeostasis. Here, we investigated the energetic cost of living in nitrate contaminated waters during early, energy-limited, larval life stages in the endangered southern bell frog (Litoria raniformis). We hypothesised that chronic nitrate exposure during development would increase energetic costs, evidenced by reductions in growth rates and body sizes, and elevations in routine heart rate (RHR) and routine metabolic rate (RMR). Following hatching, larvae were exposed to one of three nitrate treatments (0, 25 and 50 mg NO3-L-1) for 12 weeks, and survival, growth, RHR, and RMR were measured. Survival rates were similar across all treatments. Nitrate exposure caused a reduction in growth rates, resulting in larvae with significantly smaller body sizes. Compared to controls, nitrate-exposed larvae were 12% and 18% smaller in total length in the 25 and 50 mgNO3-L-1 treatments, respectively. However, RHR and RMR were independent of nitrate exposure, indicating that the 'cost of living' was similar across treatments. Observed growth reductions were therefore independent of RHR and RMR, suggesting other mechanisms were involved. Taken together, these results highlight the vulnerability of L. raniformis to nitrate during early life and suggest that the application of nitrogen-based fertilizers near critical aquatic habitats will be harmful.

The mechanistic basis and adaptive significance of cross-tolerance: a 'pre-adaptation' to a changing world?

Protective responses are pivotal in aiding organismal persistence in complex, multi-stressor environments. Multiple-stressor research has traditionally focused on the deleterious effects of exposure to concurrent stressors. However, encountering one stressor can sometimes confer heightened tolerance to a second stressor, a phenomenon termed 'cross-protection'. Cross-protection has been documented in a wide diversity of taxa (spanning the bacteria, fungi, plant and animal kingdoms) and habitats (intertidal, freshwater, rainforests and polar zones) in response to many stressors (e.g. hypoxia, predation, desiccation, pathogens, crowding, salinity, food limitation). Remarkably, cross-protection benefits have also been shown among emerging, anthropogenic stressors, such as heatwaves and microplastics. In this Commentary, we discuss the mechanistic basis and adaptive significance of cross-protection, and put forth the idea that cross-protection will act as a 'pre-adaptation' to a changing world. We highlight the critical role that experimental biology has played in disentangling stressor interactions and provide advice for enhancing the ecological realism of laboratory studies. Moving forward, research will benefit from a greater focus on quantifying the longevity of cross-protection responses and the costs associated with this protective response. This approach will enable us to make robust predictions of species' responses to complex environments, without making the erroneous assumption that all stress is deleterious.

Metabolic consequences of sex reversal in two lizard species: a test of the like-genotype and like-phenotype hypotheses.

Vertebrate sex is typically determined genetically, but in many ectotherms sex can be determined by genes (genetic sex determination, GSD), temperature (temperature-dependent sex determination, TSD), or interactions between genes and temperature during development. TSD may involve GSD systems with either male or female heterogamety (XX/XY or ZZ/ZW) where temperature overrides chromosomal sex determination to cause a mismatch between genetic sex and phenotypic sex (sex reversal). In these temperature-sensitive lineages, phylogenetic investigations point to recurrent evolutionary shifts between genotypic and temperature-dependent sex determination. These evolutionary transitions in sex determination can occur rapidly if selection favours the reversed sex over the concordant phenotypic sex. To investigate the consequences of sex reversal on offspring phenotypes, we measured two energy-driven traits (metabolism and growth) and 6 month survival in two species of reptile with different patterns of temperature-induced sex reversal. Male sex reversal occurs in Bassiana duperreyi when chromosomal females (female XX) develop male phenotypes (maleSR XX), while female sex reversal occurs in Pogona vitticeps when chromosomal males (male ZZ) develop female phenotypes (femaleSR ZZ). We show metabolism in maleSR XX was like that of male XY; that is, reflective of phenotypic sex and lower than genotypic sex. In contrast, for Pogona vitticeps, femaleSR ZZ metabolism was intermediate between male ZZ and female ZW metabolic rate. For both species, our data indicate that differences in metabolism become more apparent as individuals become larger. Our findings provide some evidence for an energetic advantage from sex reversal in both species but do not exclude energetic processes as a constraint on the distribution of sex reversal in nature.

Stress history affects heat tolerance in an aquatic ectotherm (Chinook salmon, Oncorhynchus tshawytscha).

The stress history of an ectotherm may be a pivotal predictor of how they cope with rapid spikes in environmental temperature. An understanding of how stressors in habitats and commercial operations affect ectotherm heat tolerance is urgently required so that management actions can be informed by thermal physiology. We hypothesised that brief exposure to mild stress would heighten tolerance to subsequent heat stress, indicative of a cross-tolerance interaction, whereas exposure to severe stress would reduce heat tolerance, reflecting a cross-susceptibility interaction. To test this hypothesis, we assessed how three acute stressors (salinity shock [10 or 33 ppt for 2 h]), air exposure (1 or 5 min) and crowding [95.6 kg m-3 for 2 h]), commonly experienced by fish, affected the heat tolerance (measured as critical thermal maximum, CTMAX) in juvenile Chinook salmon (Oncorhynchus tshawytscha). Fish were exposed to one of the three stressors and left for 24 h of recovery prior to measuring CTMAX. Heat tolerance was improved by ∼0.6 °C in fish exposed to salinity shock (10 ppt) and air exposure (5 min) compared to unstressed controls, demonstrating cross-tolerance. However, the development of cross-tolerance was non-linear with stressor severity, and crowding stress had no effect on CTMAX. Together these results show that some forms of stress can heighten acute heat tolerance in ectotherms, but the development of cross-tolerance is highly specific to both stressor type and stressor severity.

Exercise training does not affect heat tolerance in Chinook salmon (Oncorhynchus tshawytscha).

The progression of climate warming will expose ectotherms to transient heatwave events and temperatures above their tolerance range at increased frequencies. It is therefore pivotal that we understand species' physiological limits and the capacity for various controls to plastically alter these thresholds. Exercise training could have beneficial impacts on organismal heat tolerance through improvements in cardio-respiratory capacity, but this remains unexplored. Using juvenile Chinook salmon (Oncorhynchus tshawytscha), we tested the hypothesis that exercise training improves heat tolerance through enhancements in oxygen-carrying capacity. Fish were trained once daily at 60% of their maximum sustainable swim speed, UCRIT, for 60 min. Tolerance to acute warming was assessed following three weeks of exercise training, measured as the critical thermal maximum (CTMAX). CTMAX measurements were coupled with examinations of the oxygen carrying capacity (haematocrit, haemoglobin concentration, relative ventricle size, and relative splenic mass) as critical components of the oxygen transport cascade in fish. Contrary to our hypothesis, we found that exercise training did not raise the CTMAX of juvenile Chinook salmon with a mean CTMAX increase of just 0.35 °C compared to unexercised control fish. Training also failed to improve the oxygen carrying capacity of fish. Exercise training remains a novel strategy against acute warming that requires substantial fine-tuning before it can be applied to the management of commercial and wild fishes.

Adding climate change to the mix: responses of aquatic ectotherms to the combined effects of eutrophication and warming.

The threat of excessive nutrient enrichment, or eutrophication, is intensifying across the globe as climate change progresses, presenting a major management challenge. Alterations in precipitation patterns and increases in temperature are increasing nutrient loadings in aquatic habitats and creating conditions that promote the proliferation of cyanobacterial blooms. The exacerbating effects of climate warming on eutrophication are well established, but we lack an in-depth understanding of how aquatic ectotherms respond to eutrophication and warming in tandem. Here, I provide a brief overview and critique of studies exploring the cumulative impacts of eutrophication and warming on aquatic ectotherms, and provide forward direction using mechanistically focused, multi-threat experiments to disentangle complex interactions. Evidence to date suggests that rapid warming will exacerbate the negative effects of eutrophication on aquatic ectotherms, but gradual warming will induce physiological remodelling that provides protection against nutrients and hypoxia. Moving forward, research will benefit from a greater focus on unveiling cause and effect mechanisms behind interactions and designing treatments that better mimic threat dynamics in nature. This approach will enable robust predictions of species responses to ongoing eutrophication and climate warming and enable the integration of climate warming into eutrophication management policies.

Diving in hot water: a meta-analytic review of how diving vertebrate ectotherms will fare in a warmer world.

Diving ectothermic vertebrates are an important component of many aquatic ecosystems, but the threat of climate warming is particularly salient to this group. Dive durations typically decrease as water temperatures rise; yet, we lack an understanding of whether this trend is apparent in all diving ectotherms and how this group will fare under climate warming. We compiled data from 27 studies on 20 ectothermic vertebrate species to quantify the effect of temperature on dive durations. Using meta-analytic approaches, we show that, on average, dive durations decreased by 11% with every 1°C increase in water temperature. Larger increases in temperature (e.g. +3°C versus +8-9°C) exerted stronger effects on dive durations. Although species that respire bimodally are projected to be more resilient to the effects of temperature on dive durations than purely aerial breathers, we found no significant difference between these groups. Body mass had a weak impact on mean dive durations, with smaller divers being impacted by temperature more strongly. Few studies have examined thermal phenotypic plasticity (N=4) in diving ectotherms, and all report limited plasticity. Average water temperatures in marine and freshwater habitats are projected to increase between 1.5 and 4°C in the next century, and our data suggest that this magnitude of warming could translate to substantial decreases in dive durations, by approximately 16-44%. Together, these data shed light on an overlooked threat to diving ectothermic vertebrates and suggest that time available for underwater activities, such as predator avoidance and foraging, may be shortened under future warming.

Harnessing the potential of cross-protection stressor interactions for conservation: a review.

Conservation becomes increasingly complex as climate change exacerbates the multitude of stressors that organisms face. To meet this challenge, multiple stressor research is rapidly expanding, and the majority of this work has highlighted the deleterious effects of stressor interactions. However, there is a growing body of research documenting cross-protection between stressors, whereby exposure to a priming stressor heightens resilience to a second stressor of a different nature. Understanding cross-protection interactions is key to avoiding unrealistic 'blanket' conservation approaches, which aim to eliminate all forms of stress. But, a lack of synthesis of cross-protection interactions presents a barrier to integrating these protective benefits into conservation actions. To remedy this, we performed a review of cross-protection interactions among biotic and abiotic stressors within a conservation framework. A total of 66 publications were identified, spanning a diverse array of stressor combinations and taxonomic groups. We found that cross-protection occurs in response to naturally co-occurring stressors, as well as novel, anthropogenic stressors, suggesting that cross-protection may act as a 'pre-adaptation' to a changing world. Cross-protection interactions occurred in response to both biotic and abiotic stressors, but abiotic stressors have received far more investigation. Similarly, cross-protection interactions were present in a diverse array of taxa, but several taxonomic groups (e.g. mammals, birds and amphibians) were underrepresented. We conclude by providing an overview of how cross-protection interactions can be integrated into conservation and management actions and discuss how future research in this field may be directed to improve our understanding of how cross-protection may shield animals from global change.

Double whammy: Nitrate pollution heightens susceptibility to both hypoxia and heat in a freshwater salmonid.

Species persistence in a changing world will depend on how they cope with co-occurring stressors. Stressors can interact in unanticipated ways, where exposure to one stressor may heighten or reduce resilience to another stressor. We examined how a leading threat to aquatic species, nitrate pollution, affects susceptibility to hypoxia and heat stress in a salmonid, the European grayling (Thymallus thymallus). Fish were exposed to nitrate pollution (0, 50 or 200 mg NO3- L-1) at two acclimation temperatures (18 °C or 22 °C) for eight weeks. Hypoxia- and heat-tolerance were subsequently assessed, and the gills of a subset of fish were sampled for histological analyses. Nitrate-exposed fish were significantly more susceptible to acute hypoxia at both acclimation temperatures. Similarly, in 18 °C- acclimated fish, exposure to 200 mg NO3- L- 1 caused a 1 °C decrease in heat tolerance (critical thermal maxima, CTMax). However, the opposite effect was observed in 22 °C-acclimated fish, where nitrate exposure increased heat tolerance by ~1 °C. Further, nitrate exposure induced some histopathological changes to the gills, which limit oxygen uptake. Our findings show that nitrate pollution can heighten the susceptibility of fish to additional threats in their habitat, but interactions are temperature dependent.

Aerobic scope and climate warming: Testing the "plastic floors and concrete ceilings" hypothesis in the estuarine crocodile (Crocodylus porosus).

Ectotherms are predicted to show a reduction in absolute aerobic scope (AAS = maximum - standard metabolic rates) if habitat temperatures surpass optima. However, thermal phenotypic plasticity may play a protective role in the maintenance of AAS. In fishes, resting physiological rates ("physiological floors," e.g., standard metabolic rates [SMR]) are typically thermally phenotypically plastic whilst maximum physiological rates ("physiological ceilings," e.g., maximum metabolic rate [MMR]) are typically fixed. This observation led to the "plastic floors and concrete ceilings" hypothesis. The applicability of this hypothesis to nonavian reptiles remains untested, despite this group being at risk of climate warming-induced extinction. We tested this hypothesis in juvenile estuarine crocodiles (Crocodylus porosus) by maintaining animals at a water temperature indicative of current summer conditions (28°C) or at a water temperature reflecting a high magnitude of warming (34°C; i.e., thermal acclimation treatments) for 6 months. Metabolic traits (SMR, MMR, and AAS) were subsequently quantified between 28-36°C. A twofold increase in SMR was observed between 28°C and 36°C in both thermal acclimation treatments (pooled Q10 = 3.2). MMR was thermally insensitive between 28°C and 36°C in 28°C-acclimated crocodiles but doubled between 28°C and 36°C in 34°C-acclimated crocodiles. These findings demonstrate thermal phenotypic plasticity in a "physiological ceiling" (MMR) and rigidity in a "physiological floor" (SMR), showing the opposite pattern to many fishes. Overall, crocodiles displayed impressive aerobic capacity at temperatures reflecting climate warming scenarios. AAS remained unchanged across an 8°C temperature range in 28°C-acclimated animals and doubled in 34°C-acclimated animals.

Synergism between elevated temperature and nitrate: Impact on aerobic capacity of European grayling, Thymallus thymallus in warm, eutrophic waters.

Climate warming and nitrate pollution are pervasive aquatic stressors that endanger the persistence of fishes prevailing in anthropogenically disturbed habitats. Individually, elevated nitrate and temperature can influence fish energy homeostasis by increasing maintenance costs and impairing oxygen transport capacity. However, it remains unknown how fish respond to simultaneous exposure to elevated temperature and nitrate pollution. Hence, we examined the combined effects of nitrate and elevated temperatures on aerobic scope (AS, maximum-standard metabolic rates) and cardiorespiratory attributes (haemoglobin HB, haematocrit HCT, relative ventricle mass RVM, and somatic spleen index SSI) in a freshwater salmonid, Thymallus thymallus. A 3 × 2 factorial design was used, where fish were exposed to one of three ecologically relevant levels of nitrate (0, 50, or 200 mg NO3- l-1) and one of two temperatures (18 °C or 22 °C) for 6 weeks. Elevated temperature increased AS by 36 % and the improvement was stronger when coupled with nitrate exposure, indicating a positive synergistic interaction. HB was reduced by nitrate exposure, while HCT was independent of nitrate pollution and temperature. Stressor exposure induced remodeling of key elements of the cardiorespiratory system. RVM was 39 % higher in fish exposed to 22 °C compared to 18 °C but was independent of nitrate exposure. SSI was independent of temperature but was 85 % and 57 % higher in fish exposed to 50 and 200 mg NO3- l-1, respectively. Taken together, these results highlight that simultaneous exposure to elevated temperatures and nitrate pollution offers cross-tolerance benefits, which may be underscored by cardiorespiratory remodeling.

Nitrite-induced reductions in heat tolerance are independent of aerobic scope in a freshwater teleost.

Nitrite is a widespread form of pollution that directly lowers the blood oxygen carrying capacity of aquatically respiring species. It is unknown if this impairment of oxygen transport translates into an increased susceptibility to elevated temperatures. We hypothesised that nitrite exposure would lower blood oxygen carrying capacity and decrease both aerobic scope (maximum-standard metabolic rate) and heat tolerance. To test these hypotheses, juvenile European carp (Cyprinus carpio) were exposed to two levels of nitrite (0 mmol l-1 or 1 mmol l-1) for 7 days and haematological parameters, critical thermal maxima (CTmax) and aerobic scope were assessed. Nitrite exposure reduced total haemoglobin by 32.9%. Aerobic scope remained unchanged in fish exposed to nitrite; however, marked declines in CTmax (1.2°C reduction) were observed in nitrite-exposed fish. These findings demonstrate that nitrite exposure can significantly impair heat tolerance, even when aerobic capacity is maintained.

Integrating physiological data with the conservation and management of fishes: a meta-analytical review using the threatened green sturgeon (Acipenser medirostris).

Reversing global declines in the abundance and diversity of fishes is dependent on science-based conservation solutions. A wealth of data exist on the ecophysiological constraints of many fishes, but much of this information is underutilized in recovery plans due to a lack of synthesis. Here, we used the imperiled green sturgeon (Acipenser medirostris) as an example of how a quantitative synthesis of physiological data can inform conservation plans, identify knowledge gaps and direct future research actions. We reviewed and extracted metadata from peer-reviewed papers on green sturgeon. A total of 105 publications were identified, spanning multiple disciplines, with the primary focus being conservation physiology (23.8%). A meta-analytical approach was chosen to summarize the mean effects of prominent stressors (elevated temperatures, salinity, low food availability and contaminants) on several physiological traits (growth, thermal tolerance, swimming performance and heat shock protein expression). All examined stressors significantly impaired green sturgeon growth, and additional stressor-specific costs were documented. These findings were then used to suggest several management actions, such as mitigating salt intrusion in nursery habitats and maintaining water temperatures within optimal ranges during peak spawning periods. Key data gaps were also identified; research efforts have been biased towards juvenile (38.1%) and adult (35.2%) life-history stages, and less data are available for early life-history stages (embryonic, 11.4%; yolk-sac larvae, 12.4%; and post yolk-sac larvae, 16.2%). Similarly, most data were collected from single-stressor studies (91.4%) and there is an urgent need to understand interactions among stressors as anthropogenic change is multi-variate and dynamic. Collectively, these findings provide an example of how meta-analytic reviews are a powerful tool to inform management actions, with the end goal of maximizing conservation gains from research efforts.

Effect of water pH and calcium on ion balance in five fish species of the Mekong Delta.

Acidic freshwater habitats disrupt ion-homeostasis in fishes, yet the often acidic waters of the Mekong host the second highest diversity of freshwater fish in the world. To investigate how five Mekong fish species tolerate water acidity, we measured: time to loss of equilibrium (LOE) at sustained (4 days) low pH (3.5) and net ion flux in acute low pH (3.5 and 3) in Chitala ornata, Pangasianodon hypophthalmus, Osphronemus goramy, Trichogaster pectoralis, and Monopterus albus. Our sustained low pH exposures revealed that C. ornata was least tolerant, P. hypophthalmus and M. albus were moderately tolerant, and O. goramy and T. pectoralis were highly tolerant to low pH. In general, net ion loss in acute low pH exposures was greatest in species with the shortest time to LOE in the sustained low pH exposure. We also explored how low water [Ca2+] (relative to current Mekong levels) affected ion flux at low water pH in the least tolerant C. ornata and highly tolerant T. pectoralis. In C. ornata, low water Ca2+ (56 ±â€¯1 µmol L-1) increased net ion loss relative to high Ca2+ (342 ±â€¯3 µmol L-1) water while no effect was observed in T. pectoralis. Finally, we find that T. pectoralis is among the most acid-tolerant fish species examined to date.

Diving beyond Aerobic Limits: Effect of Temperature on Anaerobic Support of Simulated Predator Avoidance Dives in an Air-Breathing Ectotherm.

Diving optimality models predict air breathers to routinely dive within aerobic limits, but predator avoidance dives may be an exception. Lengthening submergence times during a predation threat may enhance survival probability, and we therefore hypothesized that predator avoidance dives in juvenile estuarine crocodiles (Crocodylus porosus) would be partially anaerobically fueled. We also predicted that reliance on anaerobic metabolism would increase at elevated temperatures to offset the faster depletion of body oxygen stores. Crocodiles were maintained at 28° and 34°C for 60 d and subsequently underwent simulated predator avoidance dive trials at two test temperatures (28° and 34°C). Blood was sampled immediately on surfacing to measure plasma lactate concentrations relative to nondiving (control) values. Aerobic dive limits (cADL; min) were also calculated using known body mass and oxygen storage relationships and rates of diving oxygen consumption and compared with observed dive durations. Postdive plasma lactate levels were elevated beyond resting levels at both test temperatures, indicating that aerobic thresholds were surpassed during simulated predator avoidance dives. Similarly, ≥90% of dive durations exceeded cADLs at both test temperatures. Postdive plasma lactate concentrations were independent of water temperature and thermal acclimation treatment. Together, these findings suggest that reliance on anaerobiosis during simulated predator avoidance dives is important regardless of temperature.

Plastic responses to diel thermal variation in juvenile green sturgeon, Acipenser medirostris.

Human-induced thermal variability can disrupt energy balance and performance in ectotherms; however, phenotypic plasticity may play a pivotal protective role. Ectotherm performance can be maintained in thermally heterogeneous habitats by reducing the thermal sensitivity of physiological processes and concomitant performance. We examined the capacity of juvenile green sturgeon (Acipenser medirostris) to respond to daily thermal variation. Juveniles (47 days post-hatch) were exposed to either stable (15 ±â€¯0.5 °C) or variable (narrowly variable: 13-17 °C day-1 or widely variable 11-21 °C day-1) thermoperiod treatments, with equivalent mean temperatures (15 ±â€¯0.5 °C), for 21 days. Growth (relative growth rate, % body mass gain), upper thermal tolerance (critical thermal maxima, CTMax) and the thermal sensitivity of swimming performance (critical swimming speed, Ucrit) were assessed in fish from all treatments. Accelerated growth was observed in fish maintained under widely variable temperatures compared to narrowly variable and stable temperatures. No significant variation in CTMax was observed among thermoperiod treatments, suggesting all treatment groups acclimated to the mean temperature rather than daily maximums. The widely variable treatment induced a plastic response in swimming performance, where Ucrit was insensitive to temperature and performance was maintained across a widened thermal breadth. Maximum Ucrit attained was similar among thermoperiod treatments, but performance was maximised at different test temperatures (stable: 4.62 ±â€¯0.44 BL s-1 at 15 °C; narrowly variable: 4.52 ±â€¯0.23 BL s-1 at 21 °C; widely variable: 3.90 ±â€¯0.24 BL s-1 at 11 °C, mean ±â€¯s.e.m.). In combination, these findings suggest juvenile A. medirostris are resilient to daily fluctuations in temperature, within the temperature range tested here.

Physiological mechanisms constraining ectotherm fright-dive performance at elevated temperatures.

Survival of air-breathing, diving ectotherms is dependent on their capacity to optimise the time available for obligate underwater activities, such as predator avoidance. Submergence times are thermally sensitive, with dive durations significantly reduced by increases in water temperature, deeming these animals particularly vulnerable to the effects of climate change. The physiological mechanisms underlying this compromised performance are unclear but are hypothesised to be linked to increased oxygen demand and a reduced capacity for metabolic depression at elevated temperatures. Here, we investigated how water temperature (both acute and chronic exposures) affected the physiology of juvenile estuarine crocodiles (Crocodylus porosus) performing predator avoidance dives (i.e. fright-dives). Diving oxygen consumption, 'fright' bradycardia, haematocrit and haemoglobin (indicators of blood oxygen carrying capacity) were assessed at two test temperatures, reflective of different climate change scenarios (i.e. current summer water temperatures, 28°C, and 'high' climate warming, 34°C). Diving oxygen consumption rate increased threefold between 28 and 34°C (Q10=7.4). The capacity to depress oxygen demand was reduced at elevated temperatures, with animals lowering oxygen demand from surface levels by 52.9±27.8% and 27.8±16.5% (means±s.e.m.) at 28°C and 34°C, respectively. Resting and post-fright-dive haematocrit and haemoglobin were thermally insensitive. Together these findings suggest decrements in fright-dive performance at elevated temperatures stem from increased oxygen demand coupled with a reduced capacity for metabolic depression.