Warming of aquatic ecosystems is transforming the distribution, phenology and growth of the organisms dependent upon these ecosystems. Aquatic insects such as stoneflies are especially vulnerable to warming because the aquatic nymph stage of their life cycle depends on cool, well-oxygenated, flowing water habitat. We tracked thermal effects on available aerobic capacity of the aquatic nymph stage of an iconic and vulnerable stonefly species, the giant salmonfly (Pteronarcys californica), to compare habitat thermal regime measurements for two salmonfly populations from habitats separated by a gradient in summer weekly maximum temperatures. Contrary to expectations, the thermal optima range of the warmer habitat population was cooler than for the cooler habitat population. We posit that this unexpected interpopulation variation in thermal response is more strongly driven by diel and seasonal thermal variability than by the highest summer temperatures experienced within respective habitats. Additionally, we show that summer daily maximum temperatures could result in periodic limits in available aerobic capacity to support work of the warmer habitat nymphs and may be the mechanism underlying reduced abundance relative to the upstream cooler habitat population. Our findings provide insight into potential thermal and metabolic mechanisms that could regulate the success of ecological and culturally important aquatic insect species experiencing global change. We conclude that thermal regimes and thermal variation, not just mean and maximum temperatures, are critical drivers of aquatic insect responses to water temperatures.

Hyperoxia has been shown to expand the aerobic capacity of some fishes, although there have been very few studies examining the underlying mechanisms and how they vary across different exposure durations. Here, we investigated the cardiorespiratory function of yellowtail kingfish (Seriola lalandi) acutely (~20 h) and chronically (3-5 weeks) acclimated to hyperoxia (~200% air saturation). Our results show that the aerobic performance of kingfish is limited in normoxia and increases with environmental hyperoxia. The aerobic scope was elevated in both hyperoxia treatments driven by a ~33% increase in maximum O2 uptake (MO2max), although the mechanisms differed across treatments. Fish acutely transferred to hyperoxia primarily elevated tissue O2 extraction, while increased stroke volume-mediated maximum cardiac output was the main driving factor in chronically acclimated fish. Still, an improved O2 delivery to the heart in chronic hyperoxia was not the only explanatory factor as such. Here, maximum cardiac output only increased in chronic hyperoxia compared with normoxia when plastic ventricular growth occurred, as increased stroke volume was partly enabled by an ~8%-12% larger relative ventricular mass. Our findings suggest that hyperoxia may be used long term to boost cardiorespiratory function potentially rendering fish more resilient to metabolically challenging events and stages in their life cycle.

Zebrafish have become a widely used vertebrate model in physiology and reliable measures of their metabolic rate are needed. We have developed a 3D-printed respirometer and swim tunnel system and used it for obtaining accurate measurement of standard metabolic rate (SMR) and maximal, aerobic metabolic rate (MMR) in zebrafish under rest and maximal exercise, respectively. We compared a slow (stepwise) protocol to a fast (continuous) protocol for determining MMR. The fast protocol yielded slightly (but not significantly) higher oxygen consumption rates than the slow protocol and the data, in contrast to the slow protocol, followed a normal distribution. These findings point to the fast protocol as a fast and reliable method for obtaining accurate values of MMR in zebrafish. We make the 3D drawings for printing the system available to researchers, to help streamline the field of metabolic research in zebrafish and other smaller fish species.

Female Pacific salmon often experience higher mortality than males during their once-in-a-lifetime up-river spawning migration, particularly when exposed to secondary stressors (e.g. high temperatures). However, the underlying mechanisms remain unknown. One hypothesis is that female Pacific salmon hearts are more oxygen-limited than those of males and are less able to supply oxygen to the body\'s tissues during this demanding migration. Notably, female hearts have higher coronary blood flow, which could indicate a greater reliance on this oxygen source. Oxygen limitations can develop from naturally occurring coronary blockages (i.e. coronary arteriosclerosis) found in mature salmon hearts. If female hearts rely more heavily on coronary blood flow but experience similar arteriosclerosis levels as males, they will have disproportionately impaired aerobic performance. To test this hypothesis, we measured resting (RMR) and maximum metabolic rate (MMR), aerobic scope (AS) and acute upper thermal tolerance in coho salmon (Oncorhynchus kisutch) with an intact or artificially blocked coronary oxygen supply. We also assessed venous blood oxygen and chemistry (cortisol, ions and metabolite concentrations) at different time intervals during recovery from exhaustive exercise. We found that coronary blockage impaired MMR, AS and the partial pressure of oxygen in venous blood (PvO2) during exercise recovery but did not differ between sexes. Coronary ligation lowered acute upper thermal tolerance by 1.1°C. Although we did not find evidence of enhanced female reliance on coronary supply, our findings highlight the importance of coronary blood supply for mature wild salmon, where migration success may be linked to cardiac performance, particularly during warm water conditions.

Green sturgeon (Acipenser medirostris) are an anadromous threatened species of sturgeon found along the Pacific coast of North America. The southern distinct population segment only spawns in the Sacramento River and is exposed to water temperatures kept artificially cold for the conservation and management of winter-run Chinook salmon (Oncorhynchus tshawytscha). Past research has demonstrated costs of cold-water rearing including reduced growth rates, condition and survivorship of juvenile green sturgeon. Our research investigates how the stressors of water temperature and food limitation influence the metabolic performance of green sturgeon. We reared green sturgeon at two acclimation temperatures (13 and 19°C) and two ration amounts (100% and 40% of optimal feed). We then measured the routine and maximum metabolic rates (RMR and MMR, respectively) of sturgeon acclimated to these rearing conditions across a range of acute temperature exposures (11 to 31°C). Among both temperature acclimation treatments (13 or 19°C), we found that feed restriction reduced RMR across a range of acute temperatures. The influence of feed restriction on RMR and MMR interacted with acclimation temperature. Fish reared at 13°C preserved their MMR and aerobic scope (AS) despite feed restriction, while fish fed reduced rations and acclimated to 19°C showed reduced MMR and AS capacity primarily at temperatures below 16°C. The sympatry of threatened green sturgeon with endangered salmonids produces a conservation conflict, such that cold-water releases for the conservation of at-risk salmonids may constrain the metabolic performance of juvenile green sturgeon. Understanding the impacts of environmental conditions (e.g. temperature, dissolved oxygen) on ecological interactions of green sturgeon will be necessary to determine the influence of salmonid-focused management.

Lumpfish (Cyclopterus lumpus) mortalities have been reported during the summer at some North Atlantic salmon cage-sites where they serve as \"cleaner fish.\" To better understand this species\' physiology and whether limitations in their metabolic capacity and thermal tolerance can explain this phenomenon, we compared the aerobic scope (AS) of 6°C-acclimated lumpfish (~50 g and 8.8 cm in length at the beginning of experiments) when all individuals (N = 12) were given a chase to exhaustion, a critical swim speed (Ucrit) test, and a critical thermal maximum (CTMax) test (rate of warming 2°C h-1). The Ucrit and CTMax of the lumpfish were 2.36 ± 0.08 body lengths per second and 20.6 ± 0.3°C. The AS of lumpfish was higher during the Ucrit test (206.4 ± 8.5 mg O2 kg-1 h-1) versus that measured in either the CTMax test or after the chase to exhaustion (141.0 ± 15.0 and 124.7 ± 15.5 mg O2 kg-1 h-1, respectively). Maximum metabolic rate (MMR), AS, and \"realistic\" AS (ASR) measured using the three different protocols were not significantly correlated, indicating that measurements of metabolic capacity using one of these methods cannot be used to estimate values that would be obtained using another method. Additional findings include that (1) the lumpfish\'s metabolic capacity is comparable to that of Atlantic cod, suggesting that they are not as \"sluggish\" as previously suggested in the literature, and (2) their CTMax (20.6°C when acclimated to 6°C), in combination with their recently determined ITMax (20.6°C when acclimated to 10°C), indicates that high sea-cage temperatures are unlikely to be the primary cause of lumpfish mortalities at salmon sea-cages during the summer.

Fishing causes direct removal of individuals from wild populations but can also cause a physiological disturbance in fish that are released or discarded after capture. While sublethal physiological effects of fish capture have been well studied in commercial and recreational fisheries, this issue has been overlooked for the ornamental fish trade, where it is common to capture fish from the wild and discard non-target species. We examined metabolic responses to capture and discard procedures in the three-striped dwarf cichlid Apistogramma trifasciata, a popular Amazonian aquarium species that nonetheless may be discarded when not a target species. Individuals (n = 34) were tagged and exposed to each of four treatments designed to simulate procedures during the capture and discard process: 1) a non-handling control; 2) netting; 3) netting +30 seconds of air exposure; and 4) netting +60 seconds of air exposure. Metabolic rates were estimated using intermittent-flow respirometry, immediately following each treatment then throughout recovery overnight. Increasing amounts of netting and air exposure caused an acute increase in oxygen uptake and decrease in available aerobic scope. In general, recovery occurred quickly, with rapid decreases in oxygen uptake within the first 30 minutes post-handling. Notably, however, male fish exposed to netting +60 seconds of air exposure showed a delayed response whereby available aerobic scope was constrained <75% of maximum until ~4-6 hours post-stress. Larger fish showed a greater initial increase in oxygen uptake post-stress and slower rates of recovery. The results suggest that in the period following discard, this species may experience a reduced aerobic capacity for additional behavioural/physiological responses including feeding, territory defence and predator avoidance. These results are among the first to examine impacts of discard practises in the ornamental fishery and suggest ecophysiological research can provide valuable insight towards increasing sustainable practises in this global trade.

Sustained swimming induces beneficial effects on growth and energy metabolism in some fish species. However, the absence of a standardized exercise regimen that guarantees an optimal response to physical activity is due to the anatomical, behavioral, and physiological differences among species, and the different conditions of tests applied, which are especially notable for the early stages of cultured species. The objective of this study was to assess the growth and metabolic responses of European sea bass submitted to continuous and moderate exercise exposure, selecting a practical swimming speed from swimming tests of groups of five fingerlings. The exercise-effects trial was carried out with 600 sea bass fingerlings (3-5 g body weight) distributed in two groups (control: voluntary swimming; exercised: under sustained swimming at 1.5 body lengths·s-1). After 6 weeks, growth parameters and proximal composition of both muscles were not altered by sustained swimming, but an increased synthetic capacity (increased RNA/DNA ratio) and more efficient use of proteins (decreased ΔN15) were observed in white muscle. The gene expression of mitochondrial proteins in white and red muscle was not affected by exercise, except for ucp3, which increased. The increase of UCP3 and Cox4 protein expression, as well as the higher COX/CS ratio of enzyme activity in white muscle, pointed out an enhanced oxidative capacity in this tissue during sustained swimming. In the protein expression of red muscle, only CS increased. All these metabolic adaptations to sustained exercise were also reflected in an enhanced maximum metabolic rate (MMR) with higher aerobic scope (AMS) of exercised fish in comparison to the non-trained fish, during a swimming test. These results demonstrated that moderate sustained swimming applied to sea bass fingerlings can improve the physical fitness of individuals through the enhancement of their aerobic capacities.

Winter cold slows ectotherm physiology, potentially constraining activities and ecological opportunities at poleward latitudes. Yet, many fishes are winter-active, facilitated by thermal compensation that improves cold performance. Conversely, winter-dormant fishes (e.g. cunner, Tautogolabrus adspersus) become inactive and non-feeding overwinter. Why are certain fishes winter-dormant? We hypothesized that winter dormancy is an adaptive behavioural response arising in poleward species that tolerate severe, uncompensated constraints of cold on their physiological performance. We predicted that below their dormancy threshold of 7--8°C, exercise and metabolic performance of cunner are greatly decreased, even after acclimation (i.e. shows above-normal, uncompensated thermal sensitivity, Q10>1-3). We measured multiple key performance metrics (e.g. C-start maximum velocity, chase swimming speed, aerobic scope) in cunner after acute exposure to 26-2°C (3°C intervals using 14°C-acclimated fish) or acclimation (5-8 weeks) to 14-2°C (3°C intervals bracketing the dormancy threshold). Performance declined with cooling, and the acute Q10 of all six performance rate metrics was significantly greater below the dormancy threshold temperature (Q10,acute8-2°C=1.5-4.9, mean=3.3) than above (Q10,acute14-8°C=1.1-1.9, mean=1.5), inferring a cold constraint. However, 2°C acclimation (temporally more relevant to seasonal cooling) improved performance, abolishing the acute constraint (Q10,acclimated8-2°C=1.4-3.0, mean=2.0; also cf. Q10,acclimated14-8°C=1.2-2.9, mean=1.7). Thus, dormant cunner show partial cold-compensation of exercise and metabolic performance, similar to winter-active species. However, responsiveness to C-start stimuli was greatly cold-constrained even following acclimation, suggesting dormancy involves sensory limitation. Thermal constraints on metabolic and exercise physiology are not significant drivers of winter dormancy in cunner. In fact, compensatory plasticity at frigid temperatures is retained even in a dormant fish.

Resource limitation can constrain energy (ATP) production, and thereby affect locomotion and behaviour such as exploration of novel environments and boldness. Consequently, ecological processes such as dispersal and interactions within and between species may be influenced by food availability. Energy metabolism, and behaviour are regulated by endocrine signalling, and may therefore be impacted by endocrine disrupting compounds (EDCs) including bisphenol A (BPA) derived from plastic manufacture and pollution. It is important to determine the impacts of these novel environmental contexts to understand how human activity alters individual physiology and behaviour and thereby populations. Our aim was to determine whether BPA exposure interacts with feeding frequency to alter metabolism and behaviour. In a fully factorial experiment, we show that low feeding frequency reduced zebrafish (Danio rerio) mass, condition, resting metabolic rates, total distance moved and speed in a novel arena, as well as anxiety indicated by the number of times fish returned to a dark shelter. However, feeding frequency did not significantly affect maximal metabolic rates, aerobic scope, swimming performance, latency to leave a shelter, or metabolic enzyme activities (citrate synthase and lactate dehydrogenase). Natural or anthropogenic fluctuation in food resources can therefore impact energetics and movement of animals with repercussions for ecological processes such as dispersal. BPA exposure reduced LDH activity and body mass, but did not interact with feeding frequency. Hence, behaviour of adult fish is relatively insensitive to disruption by BPA. However, alteration of LDH activity by BPA could disrupt lactate metabolism and signalling and together with reduction in body mass could affect size-dependent reproductive output. BPA released by plastic manufacture and pollution can thereby impact conservation and management of natural resources.