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Incorporating host-pathogen(s)-environment axes into management and conservation planning is critical to preserving species in a warming climate. However, the role pathogens play in host stress resilience remains largely unexplored in wild animal populations. We experimentally characterized how independent and cumulative stressors (fisheries handling, high water temperature) and natural infections affected the health and longevity of released wild adult sockeye salmon (Oncorhynchus nerka) in British Columbia, Canada. Returning adults were collected before and after entering the Fraser River, yielding marine- and river-collected groups, respectively (N = 185). Fish were exposed to a mild (seine) or severe (gill net) fishery treatment at collection, and then held in flow-through freshwater tanks for up to four weeks at historical (14°C) or projected migration temperatures (18°C). Using weekly nonlethal gill biopsies and high-throughput qPCR, we quantified loads of up to 46 pathogens with host stress and immune gene expression. Marine-collected fish had less severe infections than river-collected fish, a short migration distance (100 km, 5-7 days) that produced profound infection differences. At 14°C, river-collected fish survived 1-2 weeks less than marine-collected fish. All fish held at 18°C died within 4 weeks unless they experienced minimal handling. Gene expression correlated with infections in river-collected fish, while marine-collected fish were more stressor-responsive. Cumulative stressors were detrimental regardless of infections or collection location, probably due to extreme physiological disturbance. Because river-derived infections correlated with single stressor responses, river entry probably decreases stressor resilience of adult salmon by altering both physiology and pathogen burdens, which redirect host responses toward disease resistance.

Background Viruses can impose energetic demands on organisms they infect, in part by hosts mounting resistance. Recognizing that oxygen uptake reliably indicates steady-state energy consumption in all vertebrates, we comprehensively evaluated oxygen uptake and select transcriptomic messaging in sockeye salmon challenged with either a virulent rhabdovirus (IHNV) or a low-virulent reovirus (PRV). We tested three hypotheses relating to the energetic costs of viral resistance and tolerance in this vertebrate system: (1) mounting resistance incurs a metabolic cost or limitation, (2) induction of the innate antiviral interferon system compromises homeostasis, and (3) antiviral defenses are weakened by acute stress. Results IHNV infections either produced mortality within 1–4 weeks or the survivors cleared infections within 1–9 weeks. Transcription of three interferon-stimulated genes (ISGs) was strongly correlated with IHNV load but not respiratory performance. Instead, early IHNV resistance was associated with a mean 19% (95% CI = 7–31%; p = 0.003) reduction in standard metabolic rate. The stress of exhaustive exercise did not increase IHNV transcript loads, but elevated host inflammatory transcriptional signaling up to sevenfold. For PRV, sockeye tolerated high-load systemic PRV blood infections. ISG transcription was transiently induced at peak PRV loads without associated morbidity, microscopic lesions, or major changes in aerobic or anaerobic respiratory performance, but some individuals with high-load blood infections experienced a transient, minor reduction in hemoglobin concentration and increased duration of excess post-exercise oxygen consumption. Conclusions Contrary to our first hypothesis, effective resistance against life-threatening rhabdovirus infections or tolerance to high-load reovirus infections incurred minimal metabolic costs to salmon. Even robust systemic activation of the interferon system did not levy an allostatic load sufficient to compromise host homeostasis or respiratory performance, rejecting our second hypothesis that this ancient innate vertebrate antiviral defense is itself energetically expensive. Lastly, an acute stress experienced during testing did not weaken host antiviral defenses sufficiently to promote viral replication; however, a possibility for disease intensification contingent upon underlying inflammation was indicated. These data cumulatively demonstrate that fundamental innate vertebrate defense strategies against potentially life-threatening viral exposure impose limited putative costs on concurrent aerobic or energetic demands of the organism.

In recent decades, the relative proportion of female sockeye salmon (Oncorhynchus nerka) on spawning grounds of several British Columbia populations has declined. Coincident with the decline has been large changes to oceanic, estuarine, and river migration environments. Over the past 30 years, numerous telemetry tracking and laboratory studies have examined mortality of adult Fraser River sockeye salmon during ocean and freshwater migrations. We reviewed 19 studies that provided 40 situations where male and female mortality could be directly compared. Female mortality averaged 2.1 times greater than that of males and up to eightfold higher. High female mortality was also evident in migrating coho salmon (Oncorhynchus kisutch) and Chinook salmon (Oncorhynchus tshawytscha) and for sockeye salmon in other systems. Female mortality was highest when migration conditions were challenging (e.g., high or turbulent flows, high temperatures, confinement, or handling) and towards end of river migration. We review mechanisms for differential mortality, including energy exhaustion, cardiac performance, physiological stress, and immune factors. Female-specific mortality will become even more pronounced in coming years as ocean and riverine conditions continue to change.

This analysis shows good intentions in the selection of valid and precise oxygen uptake ( M˙ O2 ) measurements by retaining only slopes of declining dissolved oxygen level in a respirometer that have very high values of the coefficient of determination, r2 , are not always successful at excluding nonlinear slopes. Much worse, by potentially removing linear slopes that have low r2 only because of a low signal-to-noise ratio, this procedure can overestimate the calculation of standard metabolic rate (SMR) of the fish. To remedy this possibility, a few simple diagnostic tools are demonstrated to assess the appropriateness of a given minimum acceptable r2 , such as calculating the proportion of rejected M˙ O2 determinations, producing a histogram of the r2 values and a plot of r2 as a function of M˙ O2 . The authors offer solutions for cases when many linear slopes have low r2 . The least satisfactory but easiest to implement is lowering the minimum acceptable r2 . More satisfactory solutions involve processing (smoothing) the raw signal of dissolved oxygen as a function of time to improve the signal-to-noise ratio and the r2 s.

Intraspecific variation in key traits such as tolerance of warming can have profound effects on ecological and evolutionary processes, notably responses to climate change. The empirical evidence for three primary elements of intraspecific variation in tolerance of warming in fishes is reviewed. The first is purely mechanistic that tolerance varies across life stages and as fishes become mature. The limited evidence indicates strongly that this is the case, possibly because of universal physiological principles. The second is intraspecific variation that is because of phenotypic plasticity, also a mechanistic phenomenon that buffers individuals' sensitivity to negative impacts of global warming in their lifetime, or to some extent through epigenetic effects over successive generations. Although the evidence for plasticity in tolerance to warming is extensive, more work is required to understand underlying mechanisms and to reveal whether there are general patterns. The third element is intraspecific variation based on heritable genetic differences in tolerance, which underlies local adaptation and may define long-term adaptability of a species in the face of ongoing global change. There is clear evidence of local adaptation and some evidence of heritability of tolerance to warming, but the knowledge base is limited with detailed information for only a few model or emblematic species. There is also strong evidence of structured variation in tolerance of warming within species, which may have ecological and evolutionary significance irrespective of whether it reflects plasticity or adaptation. Although the overwhelming consensus is that having broader intraspecific variation in tolerance should reduce species vulnerability to impacts of global warming, there are no sufficient data on fishes to provide insights into particular mechanisms by which this may occur.

This study hypothesized that oxygen uptake (ṀO2 ) measured with a novel protocol of chasing rainbow trout Oncorhynchus mykiss to exhaustion inside a static respirometer while simultaneously monitoring ṀO2 (ṀO2chase ) would generate the same and repeatable peak value as when peak active ṀO2 (ṀO2active ) is measured in a critical swimming speed protocol. To reliably determine peak ṀO2chase , and compare to the peak during recovery of ṀO2 after a conventional chase protocol outside the respirometer (ṀO2rec ), this study applied an iterative algorithm and a minimum sampling window duration (i.e., 1 min based on an analysis of the variance in background and exercise ṀO2 ) to account for ṀO2 dynamics. In support of this hypothesis, peak ṀO2active (707 ± 33 mg O2 h-1 kg-1 ) and peak ṀO2chase (663 ± 43 mg O2 h-1 kg-1 ) were similar (P = 0.49) and repeatable (Pearson's and Spearman's correlation test; r ≥ 0.77; P < 0.05) when measured in the same fish. Therefore, estimates of ṀO2max can be independent of whether a fish is exhaustively chased inside a respirometer or swum to fatigue in a swim tunnel, provided ṀO2 is analysed with an iterative algorithm and a minimum but reliable sampling window. The importance of using this analytical approach was illustrated by peak ṀO2chase being 23% higher (P < 0.05) when compared with a conventional sequential interval regression analysis, whereas using the conventional chase protocol (1-min window) outside the respirometer increased this difference to 31% (P < 0.01). Moreover, because peak ṀO2chase was 18% higher (P < 0.05) than peak ṀO2rec , chasing a fish inside a static respirometer may be a better protocol for obtaining maximum ṀO2 .

Multiple stressors are commonly encountered by wild animals, but their cumulative effects are poorly understood, especially regarding infection development. We conducted a holding study with repeated gill and blood sampling to characterize the effects of cumulative stressors on infection development in adult coho salmon. Treatments included chronic thermal stress (15°C vs. 10°C) and acute gill net entanglement with an air exposure (simulating fisheries bycatch release). The potential loadings of 35 infectious agents and the expression of 17 host immune genes were quantified using high-throughput quantitative polymerase chain reaction, while host physiology was characterized with chemical analysis of blood. Temporal increases in infectious agent richness and loads were concurrent with decreased expression of immune genes in fish sampled in the river. In the laboratory, mortality was minimal in cool water regardless of fishery treatment ( 1 wk) and enhanced mortality associated with handling and biopsy (∼40% both sexes). Experimental gillnetting at high temperature further enhanced female mortality (73%). Fish held at high temperature demonstrated heavier infectious agent loads, osmoregulatory impairment, suppressed female maturation, and upregulation of inflammatory and extracellular immune genes. At high temperature, heavy Parvicapsula minibicornis loads were associated with premature mortality. Females exhibited physiological impairment from both stressors after 1 wk, and infection burdens correlated poorly with immune gene regulation compared with males. Cumulative effects of multiple stressors on female mortality are likely a function of physiological impairment and enhanced infections at high temperature.

Icefishes characteristically lack the oxygen-binding protein haemoglobin and therefore are especially reliant on cardiovascular regulation to augment oxygen transport when oxygen demand increases, such as during activity and warming. Using both in vivo and in vitro experiments, we evaluated the roles for adrenaline and adenosine, two well-established cardio- and vasoactive molecules, in regulating the cardiovascular system of the blackfin icefish, Chaenocephalus aceratus. Despite increasing cardiac contractility (increasing twitch force and contraction kinetics in isometric myocardial strip preparations) and accelerating heart rate (ƒ H ), adrenaline (5 nmol kg −1 bolus intra-arterial injection) did not significantly increase cardiac output (Q̇) in vivo because it elicited a large decrease in vascular conductance (G sys ). In contrast, and despite preliminary data suggesting a direct negative inotropic effect of adenosine on isolated atria and little effect on isolated ventricle strips, adenosine (500 nmol kg −1 ) generated a large increase in Q̇ by increasing G sys , a change reminiscent of that previously reported during both acute warming and invoked activity. Our data thus illustrate how Q̇ in C. aceratus may be much more dependent on peripheral control of vasomotor tone than direct regulation of the heart.

We show that cardiac sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) activity differs considerably among sockeye salmon (Oncorhynchus nerka) populations. Variability in SERCA activity was significantly correlated with elevation gain and temperature during migration, as well as maximum cardiac stroke volume. Furthermore, because SERCA activity was not lowered during the spawning migration, this aspect of the cardiac contraction machinery is apparently spared during the senescence of these semelparous salmon, likely because it is essential for these fish to complete spawning. Only when spawning had been completed was there a significant reduction in SERCA activity, which was detectable in males at a 25 °C and in females at a 15 °C assay temperature. Hence, we propose that migration conditions act as a strong selective force that has resulted in local adaptation of myocardial SERCA activity among sockeye salmon populations.

Warming in the region of the Western Antarctic Peninsula is occurring at an unprecedented rate, which may threaten the survival of Antarctic notothenioid fishes. Herein, we review studies characterizing thermal tolerance and cardiac performance in notothenioids – a group that includes both red-blooded species and the white-blooded, haemoglobinless icefishes – as well as the relevant biochemistry associated with cardiac failure during an acute temperature ramp. Because icefishes do not feed in captivity, making long-term acclimation studies unfeasible, we focus only on the responses of red-blooded notothenioids to warm acclimation. With acute warming, hearts of the white-blooded icefish Chaenocephalus aceratus display persistent arrhythmia at a lower temperature (8°C) compared with those of the red-blooded Notothenia coriiceps (14°C). When compared with the icefish, the enhanced cardiac performance of N. coriiceps during warming is associated with greater aerobic capacity, higher ATP levels, less oxidative damage and enhanced membrane integrity. Cardiac performance can be improved in N. coriiceps with warm acclimation to 5°C for 6–9 weeks, accompanied by an increase in the temperature at which cardiac failure occurs. Also, both cardiac mitochondrial and microsomal membranes are remodelled in response to warm acclimation in N. coriiceps, displaying homeoviscous adaptation. Overall, cardiac performance in N. coriiceps is malleable and resilient to warming, yet thermal tolerance and plasticity vary among different species of notothenioid fishes; disruptions to the Antarctic ecosystem driven by climate warming and other anthropogenic activities endanger the survival of notothenioids, warranting greater protection afforded by an expansion of marine protected areas.

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We tested in six fish species [Pacific lamprey (Lampetra richardsoni), Pacific spiny dogfish (Squalus suckleyi), Asian swamp eel (Monopterus albus), white sturgeon (Acipenser transmontanus), zebrafish (Danio rerio), and starry flounder (Platichthys stellatus)] the hypothesis that elevated extracellular [HCO3-] protects spontaneous heart rate and cardiac force development from the known impairments that severe hypoxia and hypercapnic acidosis can induce. Hearts were exposed in vitro to either severe hypoxia (~ 3% of air saturation), or severe hypercapnic acidosis (either 7.5% CO2 or 15% CO2), which reduced heart rate (in six test species) and net force development (in three test species). During hypoxia, heart rate was restored by [HCO3-] in a dose-dependent fashion in lamprey, dogfish and eel (EC50 = 5, 25 and 30mM, respectively), but not in sturgeon, zebrafish or flounder. During hypercapnia, elevated [HCO3-] completely restored heart rate in dogfish, eel and sturgeon (EC50 = 5, 25 and 30mM, respectively), had a partial effect in lamprey and zebrafish, and had no effect in flounder. Elevated [HCO3-], however, had no significant effect on net force of electrically paced ventricular strips from dogfish, eel and flounder during hypoxia and hypercapnia. Only in lamprey hearts did a specific soluble adenylyl cyclase (sAC) inhibitor, KH7, block the HCO3--mediated rescue of heart rate during both hypoxia and hypercapnia, and wasthe only species where we conclusively demonstrated sAC activity was involved in the protective effects of HCO3- on cardiac function. Our results suggest a common HCO3--dependent, sAC-dependent transduction pathway for heart rate recovery exists in cyclostomes and a HCO3--dependent, sAC-independent pathway exists in other fish species.

Both laboratory and field respirometry are rapidly growing techniques to determine animal performance thresholds. However, replicating protocols to estimate maximum metabolic rate (MMR) between species, populations, and individuals can be difficult, especially in the field. We therefore evaluated seven different exercise treatments-four laboratory methods involving a swim tunnel (critical swim speed [Ucrit], Ucrit postswim fatigue, maximum swim speed [Umax], and Umax postswim fatigue) and three field-based chasing methods (3-min chase with 1-min air exposure, 3-min chase with no air exposure, and chase to exhaustion)-in adult coho salmon (Oncorhynchus kisutch) as a case study to determine best general practices for measuring and quantifying MMR in fish. We found that all seven methods were highly comparable and that chase treatments represent a valuable field alternative to swim tunnels. Moreover, we caution that the type of test and duration of measurement windows used to calculate MMR can have significant effects on estimates of MMR and statistical power for each approach.

Adult female Pacific salmon can have higher migration mortality rates than males, particularly at warm temperatures. However, the mechanisms underlying this phenomenon remain a mystery. Given the importance of swimming energetics on fitness, we measured critical swim speed, swimming metabolism, cost of transport, aerobic scope (absolute and factorial) and exercise recovery in adult female and male coho salmon (Oncorhynchus kisutch) held for 2 days at 3 environmentally relevant temperatures (9°C, 14°C, 18°C) in fresh water. Critical swimming performance (Ucrit) was equivalent between sexes and maximal at 14°C. Absolute aerobic scope was sex- and temperature-independent, whereas factorial aerobic scope decreased with increasing temperature in both sexes. The full cost of recovery from exhaustive exercise (excess post-exercise oxygen consumption) was higher in males compared to females. Immediately following exhaustive exercise (i.e. 1 h), recovery was impaired at 18°C for both sexes. At an intermediate time scale (i.e. 5 h), recovery in males was compromised at 14°C and 18°C compared to females. Overall, swimming, aerobic metabolism, and recovery energetics do not appear to explain the phenomenon of increased mortality rates in female coho salmon. However, our results suggest that warming temperatures compromise recovery following exhaustive exercise in both male and female salmon, which may delay migration progression and could contribute to en route mortality.

Despite immense concern over amplified warming in the Arctic, physiological research to address related conservation issues for valuable cold-adapted fish, such as the Arctic char (Salvelinus alpinus), is lacking. This crucial knowledge gap is largely attributable to the practical and logistical challenges of conducting sensitive physiological investigations in remote field settings. Here, we used an innovative, mobile aquatic-research laboratory to assess the effects of temperature on aerobic metabolism and maximum heart rate (fHmax) of upriver migrating Arctic char in the Kitikmeot region of Nunavut in the central Canadian Arctic. Absolute aerobic scope was unchanged at temperatures from 4 to 16°C, while fHmax increased with temperature (Q10 = 2.1), as expected. However, fHmax fell precipitously below 4°C and it began to plateau above ~ 16°C, reaching a maximum at ~ 19°C before declining and becoming arrhythmic at ~ 21°C. Furthermore, recovery from exhaustive exercise appeared to be critically impaired above 16°C. The broad thermal range (~4–16°C) for increasing fHmax and maintaining absolute aerobic scope matches river temperatures commonly encountered by migrating Arctic char in this region. Nevertheless, river temperatures can exceed 20°C during warm events and our results confirm that such temperatures would limit exercise performance and thus impair migration in this species. Thus, unless Arctic char can rapidly acclimatize or alter its migration timing or location, which are both open questions, these impairments would likely impact population persistence and reduce lifetime fitness. As such, future conservation efforts should work towards quantifying and accounting for the impacts of warming, variable river temperatures on migration and reproductive success.

We examined cardiac pacemaker rate resetting in rainbow trout following a reciprocal temperature transfer. In the original experiment, performed in winter, 4°C-acclimated fish transferred to 12°C reset intrinsic heart rate after just 1 h (from 56.8±1.2 to 50.8±1.5 beats min−1); 12°C-acclimated fish transferred to 4°C reset intrinsic heart rate after 8 h (from 33.4±0.7 to 37.7±1.2 beats min−1). However, in a replicate experiment, performed in the summer using a different brood year, intrinsic heart rate was not reset, even after 10 weeks at a new temperature. Using this serendipitous opportunity, we compared mRNA expression changes of a suite of proteins in sinoatrial node (SAN), atrial and ventricular tissues after both 1 h and longer than 3 weeks for both experimental acclimation groups to identify those changes only associated with pacemaker rate resetting. Of the changes in mRNA expression occurring after more than 3 weeks of warm acclimation and associated with pacemaker rate resetting, we observed downregulation of NKA α1c in the atrium and ventricle, and upregulation of HCN1 in the ventricle. However, in the SAN there were no mRNA expression changes unique to the fish with pacemaker rate resetting after either 1 h or 3 weeks of warm acclimation. Thus, despite identifying changes in mRNA expression of contractile cardiac tissues, there was an absence of changes in mRNA expression directly involved with the initial, rapid pacemaker rate resetting with warm acclimation. Importantly, pacemaker rate resetting with thermal acclimation does not always occur in rainbow trout., Summary: Intrinsic heart rate resetting occurs inconsistently with temperature acclimation in rainbow trout; however, it can occur rapidly, within 1 h of warming, without changes in mRNA expression of sinoatrial node proteins.

The cardiovascular system is critical for delivering O2 to tissues. Here we examine the cardiovascular responses to progressive hypoxia in four high-altitude Andean duck species compared to four related low-altitude populations in North America, tested at their native altitude. Ducks were exposed to stepwise decreases in inspired partial pressure of O2 while we monitored heart rate, O2 consumption rate, blood O2 saturation, haematocrit (Hct), and blood haemoglobin concentration [Hb]. We calculated O2 pulse (the product of stroke volume and the arterial-venous O2 content difference), blood O2 concentration, and heart rate variability. Regardless of altitude, all eight populations maintained O2 consumption rate with minimal change in heart rate or O2 pulse, indicating that O2 consumption was maintained by either a constant arterial-venous O2 content difference (an increase in the relative O2 extracted from arterial blood) or by a combination of changes in stroke volume and the arterial-venous O2 content difference. Three high-altitude taxa (yellow-billed pintails, cinnamon teal, and speckled teal) had higher Hct and [Hb], increasing the O2 content of arterial blood, and potentially providing a greater reserve for enhancing O2 delivery during hypoxia. Hct and [Hb] between low- and high-altitude populations of ruddy duck were similar, representing a potential adaptation to diving life. Heart rate variability was generally lower in high-altitude ducks, concurrent with similar or lower heart rates than low-altitude ducks, suggesting a reduction in vagal and sympathetic tone. These unique features of the Andean ducks differ from previous observations in both Andean geese and bar-headed geese, neither of which exhibit significant elevations in Hct or [Hb] compared to their low-altitude relatives, revealing yet another avian strategy for coping with high altitude.

Fish released after capture, or fish interacting with gear but escaping, sometimes experience fishing-related incidental mortality (FRIM). For adult Pacific salmon migrations, knowing the magnitude of FRIM is important to estimate escapement accurately and to understand the total impact of a specific fishery. To determine how multiple gear types are associated with FRIM at different levels of maturity, we captured sockeye salmon (Oncorhynchus nerka) by both gill net and beach seine at three locations along their migration route (10%, 26%, and 72% of a 500 km freshwater migration) and determined their migratory success using biotelemetry. FRIM was higher for fish captured by gill net except at the location closest to spawning grounds. In addition, salmon captured by gill net at the lower river locations temporarily delayed migration, potentially indicating a requirement for lengthier recovery time compared with beach-seined fish. These results provide the first empirical and parallel comparison of these two common in-river fishing methods for salmon, revealing clear differences in FRIM between the two fishing methods in lower river fisheries and the importance of maturity.

The early life stages of Pacific salmon are at risk of environmental exposure to diluted bitumen (dilbit) as Canada’s oil sands industry continues to expand. The toxicity and latent effects of dilbit exposure were assessed in sockeye salmon (Oncorhynchus nerka) exposed to water-soluble fractions of dilbit (WSFd) from fertilization to the swim-up stage, and then reared in clean water for 8 months. Mortality was significantly higher in WSFd-exposed embryos, with cumulative mortality up to 4.6-fold higher in exposed relative to unexposed embryos. The sublethal effects of WSFd exposure included transcriptional up-regulation of cyp1a, a concentration-dependent delay in the onset and progression of hatching, as well as increased prevalence of developmental deformities at total polycyclic aromatic hydrocarbon (TPAH) concentrations ≥35 μg L−1. Growth and body composition were negatively affected by WSFd exposure, including a concentration-specific decrease in soluble protein concentration and increases in total body lipid and triglyceride concentrations. Mortality continued during the first 2 months after transferring fish to clean water, reaching 53% in fish exposed to 100 μg L−1 TPAH; but there was no latent impact on swimming performance, heart mass, or heart morphology in surviving fish after 8 months. A latent effect of WSFd exposure on brain morphology was observed, with fish exposed to 4 μg L−1 TPAH having significantly larger brains compared to other treatment groups after 8 months in clean water. This study provides comprehensive data on the acute, sub-chronic, and latent impacts of dilbit exposure in early life stage sockeye, information that is critical for a proper risk analysis of the impact of a dilbit spill on this socioeconomically important fish species.

Temperature is a master environmental factor that limits the geographical distribution of species, especially in ectotherms. To address challenges in biodiversity conservation under ongoing climate change, it is essential to characterize relevant functional limitations and adaptive genomic content at population and species levels. Here, we present evidence for adaptive divergence in cardiac function and genomic regions in redband trout (Oncorhynchus mykiss gairdneri) populations from desert and montane streams. Cardiac phenotypes of individual fish were measured in the field with a custom‐built electrocardiogram apparatus. Maximum heart rate and its rate limiting temperature during acute warming were significantly higher in fish that have evolved in the extreme of a desert climate compared to a montane climate. Association mapping with 526,301 single nucleotide polymorphisms (SNPs) across the genome revealed signatures of thermal selection both within and among ecotypes. Among desert and montane populations, 435 SNPs were identified as putative outliers under natural selection and 20 of these loci showed significant association with average summer water temperatures among populations. Phenotypes for cardiac performance were variable within each ecotype, and 207 genomic regions were strongly associated with either maximum heart rate or rate limiting temperatures among individuals. Annotation of significant loci provided candidate genes that underlie thermal adaptation, including pathways associated with cardiac function (IRX5, CASQ1, CAC1D, and TITIN), neuroendocrine system (GPR17 and NOS), and stress response (SERPH). By integrating comparative physiology and population genomics, results here advance our knowledge on evolutionary processes of thermal adaptation in aquatic ectotherms.

The recovery of oxygen uptake to the standard metabolic rate (SMR) following exhaustive chasing exercise in Atlantic salmon Salmo salar parr occurred in three phases (rapid, plateau and slow). The initial recovery phase lasted 0·7 h and contributed 16% to the total excess post-exercise oxygen consumption (EPOC). It was followed by a longer plateau phase that contributed 53% to the total EPOC. The slow recovery phase that completed recovery of SMR, which has not been reported previously, made a 31% contribution to the total EPOC. The plasticity of EPOC was demonstrated in exercise-trained fish. Exercise training increased EPOC by 39% when compared with control fish (mean ± S.E., 877·7 ± 73·1 v. 629·2 ± 53·4 mg O2 kg-1 , d.f. = 9, P

This study compared parr from three strains of rainbow trout Oncorhynchus mykiss to examine intraspecific variation in metabolic traits, hypoxia tolerance and upper thermal tolerance in this species. At the strain level, variation in absolute aerobic scope (AAS), critical oxygen level (O2crit ), incipient lethal oxygen saturation (ILOS) and critical thermal maximum (CTmax ) generally exhibited consistent differences among the strains, suggesting the possibility of functional associations among these traits. This possibility was further supported at the individual level by a positive correlation between ILOS and O2crit and a negative correlation between O2crit and AAS. These results indicate that intraspecific differences in hypoxia tolerance among strains of O. mykiss may be primarily determined by differences in the ability to maintain oxygen uptake in hypoxia and that variation in aerobic scope in normoxia probably plays a role in determining the ability of these fish to sustain metabolism aerobically as water oxygen saturation is reduced.

Heterogeneous and ever-changing thermal environments drive the evolution of populations and species, especially when extreme conditions increase selection pressure for traits influencing fitness. However, projections of biological diversity under scenarios of climate change rarely consider evolutionary adaptive potential of natural species. In this study, we tested for mechanistic evidence of evolutionary thermal adaptation among ecologically divergent redband trout populations (Oncorhynchus mykiss gairdneri) in cardiorespiratory function, cellular response and genomic variation. In a common garden environment, fish from an extreme desert climate had significantly higher critical thermal maximum (p .05) and broader optimum thermal window for aerobic scope (3°C) than fish from cooler montane climate. In addition, the desert population had the highest maximum heart rate during warming (20% greater than montane populations), indicating improved capacity to deliver oxygen to internal tissues. In response to acute heat stress, distinct sets of cardiac genes were induced among ecotypes, which helps to explain the differences in cardiorespiratory function. Candidate genomic markers and genes underlying these physiological adaptations were also pinpointed, such as genes involved in stress response and metabolic activity (hsp40, ldh-b and camkk2). These markers were developed into a multivariate model that not only accurately predicted critical thermal maxima, but also evolutionary limit of thermal adaptation in these specific redband trout populations relative to the expected limit for the species. This study demonstrates mechanisms and limitations of an aquatic species to evolve under changing environments that can be incorporated into advanced models to predict ecological consequences of climate change for natural organisms.

To evaluate whether oxygen-carrying capacity influences thermal tolerance in fishes, we reared four Chinook salmon families in present-day (+0°C) and possible future (+4°C) temperatures and assessed the response of hematocrit (Hct) to acute temperature stress. In the +4°C treatment, Hct increased above control levels when juvenile fish were exposed to their critical thermal maximum (CTmax). Conversely, no effect of temperature stress on Hct was found in the +0°C treatment. Hct was positively associated with CTmax ([Formula: see text]; [Formula: see text]), contributing to the CTmax of the +4°C treatment being significantly higher than that of the +0°C treatment (mean ± SD, [Formula: see text] and [Formula: see text], respectively). The association between CTmax and Hct found here supports the hypothesis that thermal tolerance is affected by oxygen supply to tissue. Moreover, the developmental plasticity of CTmax and Hct could represent an adaptive mechanism for salmon faced with climate change.

The utility of measuring whole-animal performance to frame the metabolic response to environmental hypoxia is well established. Progressively reducing ambient oxygen (O2) will initially limit maximum metabolic rate as a result of a hypoxemic state and ultimately lead to a time-limited, tolerance state supported by substrate-level phosphorylation when the O2 supply can no longer meet basic needs (standard metabolic rate, SMR). The metabolic consequences of declining ambient O2 were conceptually framed for fishes initially by Fry’s hypoxic performance curve, which characterizes the hypoxemic state and its consequences to absolute aerobic scope (AAS), and Hochachka’s concept of scope for hypoxic survival, which characterizes time-limited life when SMR cannot be supported by O2 supply. Yet, despite these two conceptual frameworks, the toolbox to assess whole-animal metabolic performance remains rather limited. Here, we briefly review the ongoing debate concerning the need to standardize the most commonly used assessments of respiratory performance in hypoxic fishes, namely critical O2 (the ambient O2 level below which maintenance metabolism cannot be sustained) and the incipient lethal O2 (the ambient O2 level at which a fish loses the ability to maintain upright equilibrium), and then we advance the idea that the most useful addition to the toolbox will be the limiting-O2 concentration (LOC) performance curve. Using Fry & Hart’s (1948) hypoxia performance curve concept, an LOC curve was subsequently developed as an eco-physiological framework by Neil et al. and derived for a group of fish during a progressive hypoxia trial by Claireaux and Lagardère (1999). In the present review, we show how only minor modifications to available respirometry tools and techniques are needed to generate an LOC curve for individual fish. This individual approach to the LOC curve determination then increases its statistical robustness and importantly opens up the possibility of examining individual variability. Moreover, if peak aerobic performance at a given ambient O2 level of each individual is expressed as a percentage of its AAS, the water dissolved O2 that supports 50% of the individual’s AAS (DOAAS-50) can be interpolated much like the P50 for an O2 hemoglobin dissociation curve (when hemoglobin is 50% saturated with O2). Thus, critical O2, incipient lethal O2, DOAAS-50 and P50 and can be directly compared within and across species. While an LOC curve for individual fish represents a start to an ongoing need to seamlessly integrate aerobic to anaerobic capacity assessments in a single, multiplexed respirometry trial, we close with a comparative exploration of some of the known whole-organism anaerobic and aerobic capacity traits to examine for correlations among them and guide the next steps.

When oil spills occur, behavior is the first line of defense for a fish to avoid being contaminated. We determined the avoidance threshold of the European seabass (Dicentrarchus labrax) to the water-soluble fraction (WSF) of oil using a dual-flow choice box. The results showed that a plume of 20%-diluted WSF (total polycyclic aromatic hydrocarbon [PAH] concentration: 8.54 μg L-1 ) triggered a significant avoidance response that was detected within 7.5 min of introducing WSF-contaminated water into the experimental setup. However, the ecological relevance of seabass capacity to detect and avoid WSF remains to be established. In the short term, such a response is indeed liable to reduce seabass contact time with oil-contaminated water and thus preserve their functional integrity. In the long term, however, avoidance may contribute to the displacement of a population into a possibly less auspicious environment, with consequences very similar to those of contaminant exposure, that is, disturbed population dynamics and demography. Environ Toxicol Chem 2018;37:854-859. © 2017 SETAC.

WOS:000411848700011; We tested the hypothesis that the chronic residual effects of an acute exposure of European sea bass C Dicentrarchus labrax') to chemically dispersed crude oil is manifest in indices of hypoxic performance rather than aerobic performance.Sea bass were pre-screened with a hypoxia challenge test to establish their incipient lethal oxygen saturation (ILOS),but on discovering a wide breadth for individual ILOS values (2.6-11.0% O-2 saturation), fish were subsequently subdivided into either hypoxia sensitive (HS) or hypoxia tolerant (HT) phenotypes,traits that were shown to be experimentally repeatable.The HT phenotype had a lower ILOS and critical oxygen saturation (O-2crit) compared with the HS phenotype and switched to glycolytic metabolism at a lower dissolved oxygen, even though both phenotypes accumulated lactate and glucose to the same plasma concentrations at ILOS. As initially hypothesized, and regardless of the phenotype considered, we found no residual effect of oil on any of the indices of aerobic performance. Contrary to our hypothesis, however, oil exposure had no residual effect on any of the indices of hypoxic performance in the HS phenotype. In the HT phenotype, on the other hand, oil exposure had residual effects as illustrated by the impaired repeatability of hypoxia tolerance and also by the 24% increase in O-2crit, the 40% increase in scope for oxygen deficit, the 17% increase in factorial scope for oxygen deficit and the 57% increase in accumulated oxygen deficit. Thus, sea bass with a HT phenotype remained chronically impaired for a minimum of 167 days following an acute 24-h oil exposure while the HS phenotypes did not. We reasoned that impaired oxygen extraction at gill due to oil exposure activates glycolytic metabolism at a higher dissolved oxygen, conferring on the HT phenotype an inferior hypoxia resistance that might eventually compromise their ability to survive hypoxic episodes.

Survival of prolonged anoxia requires a balance between cellular ATP demand and anaerobic ATP supply from glycolysis, especially in critical tissues such as the brain. To add insight into the ATP demand of the brain of the anoxia-tolerant red-eared slider turtle (Trachemys scripta) during prolonged periods of anoxic submergence, we quantified and compared the number of Na+-K+-ATPase units and their molecular activity in brain tissue from turtles acclimated to either 21°C or 5°C and exposed to either normoxia or anoxia (6h 21°C; 14days at 5°C). Na+-K+-ATPase activity and density per g tissue were similar at 21°C and 5°C in normoxic turtles. Likewise, anoxia exposure at 21°C did not induce any change in Na+-K+-ATPase activity or density. In contrast, prolonged anoxia at 5°C significantly reduced Na+-K+-ATPase activity by 55%, which was largely driven by a 50% reduction of the number of Na+-K+-ATPase units without a change in the activity of existing Na+-K+-ATPase pumps or α-subunit composition. These findings are consistent with the "channel arrest" hypothesis to reduce turtle brain Na+-K+-ATPase activity during prolonged, but not short-term anoxia, a change that likely helps them overwinter under low temperature, anoxic conditions.

Growth hormone (GH) transgenic fish have been proposed for use in aquaculture to enhance production efficiency. As part of a risk analysis for use of such fish, the influence of GH transgenesis on the potential to persist and succeed in natural ecosystems is being examined in confined laboratory conditions. GH transgenesis can greatly accelerate growth and, in culture conditions, is associated with secondary effects such as poor swimming capacity and spawning success. However, standard culture has also been shown to negatively affect fitness components of wild-type fish, raising the question of whether culture conditions influence fitness components of transgenic fish in a similar way. To examine factors influencing the phenotype of marine-stage GH transgenic salmon (T), and to determine if genotype-by-environment interactions exist at this life stage, we grew T and wild-type (NT) coho salmon ( Oncorhynchus kisutch ) over six cohort years in 350,000 L seawater tanks (termed mesocosms) designed to minimize effects of standard culture conditions. Mesocosm rearing partially facilitated development of normal size and morphology of NT fish relative to nature-reared counterparts, but altered overall body shape, indicating mesocosm conditions do not fully mimic natural environmental effects on coho salmon phenotype. T fish reared in mesocosms had larger mass at maturity than mesocosm- or nature-reared NT fish, indicating GH transgenesis can alter maximum obtainable mass in salmon. Unlike NT, T fish obtained maximum size at maturity across environments, suggesting marine environmental conditions may affect T growth less than NT growth. Screening parents for a common disease agent ( Renibacterium salmoninarum ) improved seawater survival, and T fish had lower survival than NT fish when from unscreened parents and inconsistent relative survival when from screened parents, indicating GH transgenesis may constitute an advantage or disadvantage in terms of survival. Transgenic salmon had lower swimming capacity and aerobic scope, but similar routine metabolic rate and thermal tolerance, demonstrating transgenesis can have different influences depending on what phenotype is examined. Using an alternate strain of T fish in phenotypic comparisons did not greatly influence most fitness components, although had a strong effect on female fecundity. The inconsistent influence of GH transgenesis on different fitness components, and existence of genotype-by-environment interactions during the marine life stage, complicates extrapolation of laboratory data for transgenic fish to natural environments. However, current and previous data do not provide evidence that overall increased performance of GH transgenic salmon over wild-type fish would arise in the marine environment. Statement of relevance Rearing in seawater mesocosms demonstrate that growth hormone transgenesis has inconsistent effects on marine fitness components in coho salmon.

Early marine survival of juvenile salmon is intimately associated with their physiological condition during ocean entry and especially smoltification. Smoltification is a developmental parr–smolt transformation allowing salmon to acquire the trait of seawater tolerance in preparation for marine living. Traditionally, this developmental process has been monitored using gill Na+/K+ATPase (NKA) activity or plasma hormones, but gill gene expression can be reliably used. Here, we describe the discovery of candidate genes from gill tissue for staging smoltification using comparisons of microarray studies with particular focus on the commonalities between anadromous Rainbow trout and Sockeye salmon datasets, as well as literature comparison encompassing more species. A subset of 37 candidate genes mainly from the microarray analyses was used for Taq-Man qPCR assay design and their monthly expression patterns were validated using gill samples from four groups, representing three species and two ecotypes: Coho salmon, Sockeye salmon, stream-type Chinook salmon, and ocean-type Chinook salmon. The best smoltification biomarkers, as measured by consistent changes across these four groups, were genes involved in ion regulation, oxygen transport, and immunity. Smoltification gene expression patterns (using the top 10 biomarkers) were confirmed by significant correlations with NKA activity and were associated with changes in body brightness, caudal fin darkness, and caudal peduncle length. We incorporate gene expression patterns of pre-smolt, smolt, and de-smolt trials from acute seawater transfers using a companion study to develop a preliminary seawater tolerance classification model for ocean-type Chinook salmon. This work demonstrates the potential of gene expression biomarkers to stage smoltification and classify juveniles as pre-smolt, smolt, or de-smolt.

The recent ubiquitous detection of PRV among salmonids has sparked international concern about the cardiorespiratory performance of infected wild and farmed salmon. Piscine orthoreovirus (PRV) has been shown to create substantial viremia in salmon by targeting erythrocytes for principle replication. In some instances, infections develop into heart and skeletal muscle inflammation (HSMI) or other pathological conditions affecting the respiratory system. Critical to assessing the seriousness of PRV infections are controlled infection studies that measure physiological impairment to critical life support systems. Respiratory performance is such a system and here multiple indices were measured to test the hypothesis that a low-virulence strain of PRV from Pacific Canada compromises the cardiorespiratory capabilities of Atlantic salmon. Contrary to this hypothesis, the oxygen affinity and carrying capacity of erythrocytes were unaffected by PRV despite the presence of severe viremia, minor heart pathology and transient cellular activation of antiviral response pathways. Similarly, PRV-infected fish had neither sustained nor appreciable differences in respiratory capabilities compared with control fish. The lack of functional harm to salmon infected with PRV in this instance highlights that, in an era of unprecedented virus discovery, detection of viral infection does not necessarily imply bodily harm and that viral load is not always a suitable predictor of disease within a host organism.

Female-biased mortality has been repeatedly reported in Pacific salmon during their upriver migration in both field studies and laboratory-holding experiments, especially in the presence of multiple environmental stressors, including thermal stress. Here we used coho salmon (Oncorhynchus kisutch) to test whether females exposed to elevated water temperatures (18°C) i) suppress circulating sex hormones (testosterone, 11-ketotestosterone, and estradiol), owing to elevated cortisol levels, ii) have higher activities of enzymes supporting anaerobic metabolism (e.g., lactate dehydrogenase), iii) lower activities of enzymes driving oxidative metabolism (e.g., citrate synthase; CS), in skeletal and cardiac muscle, and iv) have more oxidative stress damage and reduced capacity for antioxidant defense (lower catalase activity; CAT). We found no evidence that a higher susceptibility to oxidative stress contributes to female-biased mortality at warm temperatures. We did, however, find that females had significantly lower cardiac LDH and that 18°C significantly reduced plasma levels of testosterone and estradiol, especially in females. We also found that relative gonad size was significantly lower in the 18°C treatment regardless of sex, whereas relative liver size was significantly lower in females held at 18°C. Further, relative spleen size was significantly elevated in the 18°C treatments across both sexes, with larger warm-induced increases in females. Our results suggest that males may better tolerate bouts of cardiac hypoxia at high temperature, and that thermal stress may also disrupt testosterone- and estradiol-mediated protein catabolism, and the immune response (larger spleens) in migratory female salmon.

Here, we show that heart rate in zebrafish Danio rerio is dependent upon two pacemaking mechanisms and it possesses a limited ability to reset the cardiac pacemaker with temperature acclimation. Electrocardiogram recordings, taken from individual, anaesthetised zebrafish that had been acclimated to 18, 23 or 28°C were used to follow the response of maximum heart rate (fHmax ) to acute warming from 18°C until signs of cardiac failure appeared (up to c. 40°C). Because fHmax was similar across the acclimation groups at almost all equivalent test temperatures, warm acclimation was limited to one significant effect, the 23°C acclimated zebrafish had a significantly higher (21%) peak fHmax and reached a higher (3°C) test temperature than the 18°C acclimated zebrafish. Using zatebradine to block the membrane hyperpolarisation-activated cyclic nucleotide-gated channels (HCN) and examine the contribution of the membrane clock mechanisms to cardiac pacemaking, f Hmax was significantly reduced (by at least 40%) at all acute test temperatures and significantly more so at most test temperatures for zebrafish acclimated to 28°C vs. 23°C. Thus, HCN channels and the membrane clock were not only important, but could be modified by thermal acclimation. Using a combination of ryanodine (to block sarcoplasmic calcium release) and thapsigargin (to block sarcoplasmic calcium reuptake) to examine the contribution of sarcoplasmic reticular handling of calcium and the calcium clock, f Hmax was again consistently reduced independent of the test temperature and acclimation temperature, but to a significantly lesser degree than zatebradine for zebrafish acclimated to both 28 and 18°C. Thus, the calcium clock mechanism plays an additional role in setting pacemaker activity that was independent of temperature. In conclusion, the zebrafish cardiac pacemaker has a limited temperature acclimation ability compared with known effects for other fishes and involves two pacemaking mechanisms, one of which was independent of temperature.

Dr Wood's Commentary ([Wood, 2018][1]) provides six reasons to question the usefulness of P crit and proposes alternative Ṁ O2 versus P O2 analyses as its replacement. While we agree with some of Dr Wood's arguments, we feel that none of them warrant abandoning P crit, especially in favour of

Scaling of the heart across development can reveal the degree to which variation in cardiac morphology depends on body mass. In this study, we assessed the scaling of heart mass, left and right ventricular masses, and ventricular mass ratio, as a function of eviscerated body mass across fetal and postnatal development in Horro sheep Ovis aries (~50‐fold body mass range; N = 21). Whole hearts were extracted from carcasses, cleaned, dissected into chambers and weighed. We found a biphasic relationship when heart mass was scaled against body mass, with a conspicuous ‘breakpoint’ around the time of birth, manifest not by a change in the scaling exponent (slope), but rather a jump in the elevation. Fetal heart mass (g) increased with eviscerated body mass (M (b), kg) according to the power equation 4.90 M (b) (0.88 ± 0.26 (± 95%) (CI) ()), whereas postnatal heart mass increased according to 10.0 M (b) (0.88 ± 0.10). While the fetal and postnatal scaling exponents are identical (0.88) and reveal a clear dependence of heart mass on body mass, only the postnatal exponent is significantly less than 1.0, indicating the postnatal heart becomes a smaller component of body mass as the body grows, which is a pattern found frequently with postnatal cardiac development among mammals. The rapid doubling in heart mass around the time of birth is independent of any increase in body mass and is consistent with the normalization of wall stress in response to abrupt changes in volume loading and pressure loading at parturition. We discuss variation in scaling patterns of heart mass across development among mammals, and suggest that the variation results from a complex interplay between hard‐wired genetics and epigenetic influences.

The ecological and economic importance of fish act as a brake on the development of chemical dispersants as operational instruments following oil spills. Although a valuable and consistent body of knowledge exists, its use in spill response is limited. The objective of the present study was to increase current knowledge base to facilitate the translation of published data into information of operational value. Thus we investigated the dose-response relationship between dispersant-treated oil exposure and ecologically relevant consequences by combining laboratory and field experiments. Effects were examined over almost a year using juveniles of the slowly growing, commercially important European sea bass (Dicentrarchus labrax). A reliable interpretation of biomarker responses requires a complete knowledge of the factors likely to affect them. Interpopulational variability is of particular importance in environmental impact assessment because biomarker responses from a population collected in an impacted area are classically compared with those collected in a clean site. Our study revealed no effect of the exposure to dispersant-treated oil on fish hypoxia tolerance and temperature susceptibility at 1 and 11 mo post exposure. Similarly, no effect of the exposure was observed on the ability of the fish to cope with environmental contingencies in the field, regardless of the dose tested. Thus we feel confident to suggest that a 48-h exposure to chemically treated oil does not affect the ability of sea bass to cope with mild environmental contingencies. Finally, investigation of interpopulation variability revealed large differences in both hypoxia tolerance and temperature susceptibility among the 2 populations tested, suggesting that this variability may blur the interpretation of population comparisons as classically practiced in impact assessment. Environ Toxicol Chem 2019;38:210-221. © 2018 SETAC.

Acute warming in fish increases heart rate (fH) and cardiac output to peak values, after which performance plateaus or declines and arrhythmia may occur. This cardiac response can place a convective limitation on systemic oxygen delivery at high temperatures. To test the hypothesis that autonomic cardiac regulation protects cardiac performance in rainbow trout during acute warming, we investigated adrenergic and cholinergic regulation during the onset and progression of cardiac limitations. We explored the direct effects of adrenergic stimulation by acutely warming an in situ, working, perfused heart until arrhythmia occurred, cooling the heart to restore rhythmicity and rewarming with increasing adrenergic stimulation. Adrenergic stimulation produced a clear, dose-dependent increase in the temperature and peak fH achieved prior to the onset of arrhythmia. To examine how this adrenergic protection functions in conjunction with cholinergic vagal inhibition in vivo, rainbow trout fitted with ECG electrodes were acutely warmed in a respirometer until they lost equilibrium (CTmax) with and without muscarinic (atropine) and β-adrenergic (sotalol) antagonists. Trout exhibited roughly equal and opposing cholinergic and adrenergic tones on fH that persisted up to critical temperatures. β-adrenergic blockade significantly lowered peak fH by 14-17%, while muscarinic blockade significantly lowered the temperature for peak fH by 2.0°C. Moreover, muscarinic and β-adrenergic blockers injected individually or together significantly reduced CTmax by up to 3°C, indicating for the first time that cardiac adrenergic stimulation and cholinergic inhibition can enhance acute heat tolerance in rainbow trout at the level of the heart and the whole animal.

Increasing heart rate (ƒH) is a central, if not primary mechanism used by fishes to support their elevated tissue oxygen consumption during acute warming. Thermal acclimation can adjust this acute response to improve cardiac performance and heat tolerance under the prevailing temperatures. We predict that such acclimation will be particularly important in regions undergoing rapid environmental change such as the Arctic. Therefore, we acclimated Arctic char (Salvelinus alpinus), a high latitude, cold-adapted salmonid, to ecologically relevant temperatures (2, 6, 10, 14 and 18 °C) and examined how thermal acclimation influenced their cardiac heat tolerance by measuring the maximum heart rate (ƒHmax) response to acute warming. As expected, acute warming increased ƒHmax in all Arctic char before ƒHmax reached a peak and then became arrhythmic. The peak ƒHmax, and the temperature at which peak ƒHmax (Tpeak) and that at which arrhythmia first occurred (Tarr) all increased progressively (+33%, 49% and 35%, respectively) with acclimation temperature from 2 to 14 °C. When compared at the same test temperature ƒHmax also decreased by as much as 29% with increasing acclimation temperature, indicating significant thermal compensation. The upper temperature at which fish first lost their equilibrium (critical thermal maximum: CTmax) also increased with acclimation temperature, albeit to a lesser extent (+11%). Importantly, Arctic char experienced mortality after several weeks of acclimation at 18 °C and survivors did not have elevated cardiac thermal tolerance. Collectively, these findings suggest that if wild Arctic char have access to suitable temperatures (

The coronary circulation first appeared in the chordate lineage in cartilaginous fishes where, as in birds and mammals but unlike most teleost fishes, it supplies arterial blood to the entire myocardium. Despite the pivotal position of elasmobranch fishes in the evolution of the coronary circulation, the determinants of coronary blood flow have never been investigated in this group. Elasmobranch fishes are of special interest because of the morphological arrangement of their cardiomyocytes. Unlike teleosts, the majority of the ventricular myocardium in elasmobranch fishes is distant to the venous blood returning to the heart (i.e., the luminal blood). Also, the majority of the myocardium is in close association with the coronary circulation. To determine the relative contribution of the coronary and luminal blood supplies to cardiovascular function in sandbar sharks, Carcharhinus plumbeus, we measured coronary blood flow while manipulating cardiovascular status using acetylcholine and adrenaline. By exploring inter- and intra-individual variation in cardiovascular variables, we show that coronary blood flow is directly related to heart rate (R 2 = 0.6; P

Cardiac performance in fishes is predicted to be shaped by environmental factors such as temperature and river flow rate through natural selection for local adaptations, but few studies have explored these relationships. Using a common garden breeding design, we collected heart rate data from three populations of Atlantic salmon (Salmo salar) to measure peak heart rate and estimate optimal and upper critical temperatures for cardiac performance. We found that peak heart rate across populations matched the variation in natural river flow rates, such that the population that experienced the highest flow rate had the highest peak heart rate. Moreover, all populations showed evidence of local adaptation to summer water temperatures, with optimal temperatures (inferred from the Arrhenius breakpoint temperature) consistently falling 2.2–3.8 °C below the water temperature averaged for the summer months for each population. Also, upper critical temperatures (inferred from the temperature at which heart rates became arrhythmic) were nearly identical to peak summer water temperatures (0–0.3 °C above the peak). These results are consistent with heritable differences in cardiac performance among populations and suggest local adaptation to temperature and river flow.

Few estimates of migration rates or descriptions of behavior or survival exist for wild populations of out-migrating Pacific salmon smolts from natal freshwater rearing areas to the ocean. Using acoustic transmitters and fixed receiver arrays across four years (2010-2013), we tracked the migration of > 1850 wild sockeye salmon (Oncorhynchus nerka) smolts from Chilko Lake, British Columbia, to the coastal Pacific Ocean (> 1000 km distance). Cumulative survival to the ocean ranged 3-10% among years, although this may be slightly underestimated due to technical limitations at the final receiver array. Distinct spatial patterns in both behavior and survival were observed through all years. In small, clear, upper-river reaches, downstream migration largely occurred at night at speeds up to 50 km/d and coincided with poor survival. Among years, only 57-78% of smolts survived the first 80 km. Parallel laboratory experiments revealed excellent short-term survival and unhindered swimming performance of dummy-tagged smolts, suggesting that predators rather than tagging effects were responsible for the initial high mortality of acoustic-tagged smolts. Migration speeds increased in the Fraser River mainstem (~220 km/d in some years), diel movement patterns ceased, and smolt survival generally exceeded 90% in this segment. Marine movement rates and survival were variable across years, with among-year segment-specific survival being the most variable and lowest (19-61%) during the final (and longest, 240 km) marine migration segment. Osmoregulatory preparedness was not expected to influence marine survival, as smolts could maintain normal levels of plasma chloride when experimentally exposed to saltwater (30 ppt) immediately upon commencing their migration from Chilko Lake. Transportation of smolts downstream generally increased survival to the farthest marine array. The act of tagging may have affected smolts in the marine environment in some years as dummy-tagged fish had poorer survival than control fish when transitioned to saltwater in laboratory-based experiments. Current fisheries models for forecasting the number of adult sockeye returning to spawn have been inaccurate in recent years and generally do not incorporate juvenile or smolt survival information. Our results highlight significant potential for early migration conditions to influence adult recruitment.

The coronary circulation is of great importance in maintaining cardiovascular function and consequently it has been extensively studied in many mammalian species. However, much less attention has been paid to the coronary circulation in other vertebrates. For example, while elasmobranch fishes are of special interest as they are the most ancient lineage of vertebrates to possess a coronary circulation, only qualitative studies exist on their coronary circulation and most concern the architecture of the large arteries. Our study tested the prediction that the coronary circulation of sharks is better developed than previously thought. However, to test this idea, a methodology was needed to quantify vascularity, vessel morphology and oxygen diffusion distances in a heart with predominantly spongy myocardium. Here, we describe this methodology using dogfish and rainbow trout and suggest that the dogfish spongy myocardium appears to rely predominantly on the coronary circulation for its oxygen supply, an arrangement that contrasts with the spongy myocardial tissue of rainbow trout. In support of this suggestion, the density of the microvasculature of the spongy myocardial tissue of dogfish exceeded that of their compact tissue. Although vascularity in the compact myocardium of dogfish was significantly lower than trout, intervascular distances were similar on account of a significantly larger vessel diameter in dogfish, which corresponds to a larger red blood cell size of the dogfish when compared to trout. J. Morphol. 277:896-905, 2016. © 2016 Wiley Periodicals, Inc.

A new, image-based, tritiated ligand technique for measuring cardiac β2 -adrenoceptor (β2 -AR) binding characteristics was developed and validated with adult rainbow trout Oncorhynchus mykiss hearts so that the tissue limitation of traditional receptor binding techniques could be overcome and measurements could be made in hearts nearly 14-times smaller than previously used. The myocardial cell-surface (functional) β2 -AR density of O. nerka smolts sampled at the headwaters of the Chilko River was 54·2 fmol mg protein(-1) and about half of that previously found in return migrating adults of the same population, but still more than twice that of adult hatchery O. mykiss (21·1 fmol mg protein(-1) ). This technique now opens the possibility of investigating cardiac receptor density in a much wider range of fish species and life stages.

Oxygen supply to the heart of most teleosts, including salmonids, relies in part or in whole on oxygen-depleted venous blood. Given that plasma-accessible carbonic anhydrase (CA) in red muscle of rainbow trout has recently been shown to facilitate oxygen unloading from arterial blood under certain physiological conditions, we tested the hypothesis that plasma-accessible CA is present in the lumen of coho salmon (Oncorhynchus kisutch) hearts, and may therefore assist in the luminal oxygen supply to the spongy myocardium, which has no coronary circulation. We demonstrate a widespread distribution of CA throughout the heart chambers, including lumen-facing cells in the atrium, and confirm that the membrane-bound isoform ca4 is expressed in the atrium and ventricle of the heart. Further, we confirm that CA catalytic activity is available to blood in the atrial lumen using a modified electrometric ΔpH assay in intact atria in combination with either a membrane-impermeable CA inhibitor or specific cleavage of the Ca4 membrane anchor. Combined, these results support our hypothesis of the presence of an enhanced oxygen delivery system in the lumen of a salmonid heart, which could help support oxygen delivery when the oxygen content of venous blood becomes greatly reduced, such as after burst exercise and during environmental hypoxia.

The techniques and protocols used to successfully capture, transport and breed Arctic cod Boreogadus saida, as well as to rear their larvae through to adulthood are summarized. Breeding B. saida will increase the opportunity to study this fish species, which is a critical part of the Arctic food web.

Specific dynamic action (SDA) is the postprandial increase in oxygen uptake. Whereas it is easy to measure in fishes that remain calm and motionless during the entire digestion period, spontaneous locomotor activity is a frequent problem that leads to overestimation of SDA amplitude and magnitude (area under the curve, bound by the standard metabolic rate, SMR). Few studies have attempted to remove the effect of fish activity on SDA. A new method, non-parametric quantile regression, is described to estimate SDA even when pronounced circadian activity cycles are present. Data from juvenile Atlantic cod Gadus morhua are used to demonstrate its use and advantages compared with traditional techniques. Software (scripts in the R language) is provided to facilitate its use.

This review and data analysis outline how fish biologists should most reliably estimate the minimal amount of oxygen needed by a fish to support its aerobic metabolic rate (termed standard metabolic rate; SMR). By reviewing key literature, it explains the theory, terminology and challenges underlying SMR measurements in fishes, which are almost always made using respirometry (which measures oxygen uptake, ṀO2 ). Then, the practical difficulties of measuring SMR when activity of the fish is not quantitatively evaluated are comprehensively explored using 85 examples of ṀO2 data from different fishes and one crustacean, an analysis that goes well beyond any previous attempt. The main objective was to compare eight methods to estimate SMR. The methods were: average of the lowest 10 values (low10) and average of the 10% lowest ṀO2 values, after removing the five lowest ones as outliers (low10%), mean of the lowest normal distribution (MLND) and quantiles that assign from 10 to 30% of the data below SMR (q0·1 , q0·15 , q0·2 , q0·25 and q0·3 ). The eight methods yielded significantly different SMR estimates, as expected. While the differences were small when the variability was low amongst the ṀO2 values, they were important (>20%) for several cases. The degree of agreement between the methods was related to the c.v. of the observations that were classified into the lowest normal distribution, the c.v. MLND (C.V.MLND ). When this indicator was low (≤5·4), it was advantageous to use the MLND, otherwise, one of the q0·2 or q0·25 should be used. The second objective was to assess if the data recorded during the initial recovery period in the respirometer should be included or excluded, and the recommendation is to exclude them. The final objective was to determine the minimal duration of experiments aiming to estimate SMR. The results show that 12 h is insufficient but 24 h is adequate. A list of basic recommendations for practitioners who use respirometry to measure SMR in fishes is provided.

An organism's ability to respond effectively to environmental change is critical to its survival. Yet, life stage and overall condition can dictate tolerance thresholds to heightened environmental stressors, such that stress may not be equally felt across individuals and at all times. Also, the transcriptional responses induced by environmental changes can reflect both generalized responses as well as others that are highly specific to the type of change being experienced. Thus, if transcriptional biomarkers specific to a stressor, even under multi-stressor conditions, can be identified, the biomarkers could then be applied in natural environments to determine when and where an individual experiences such a stressor. Here, we experimentally challenged juvenile Chinook salmon (Oncorhynchus tshawytscha) to validate candidate gill gene expression biomarkers. A sophisticated experimental design manipulated salinity (freshwater, brackish water and seawater), temperature (10, 14 and 18°C) and dissolved oxygen (normoxia and hypoxia) in all 18 possible combinations for 6 days using separate trials for three smolt statuses (pre-smolt, smolt and de-smolt). In addition, changes in juvenile behaviour, plasma variables, gill Na+/K+-ATPase activity, body size, body morphology and skin pigmentation supplemented the gene expression responses. We identified biomarkers specific to salinity and temperature that transcended the multiple stressors, smolt status and mortality (live, dead and moribund). Similar biomarkers for dissolved oxygen were not identified. This work demonstrates the unique power of gene expression biomarkers to identify a specific stressor even under multi-stressor conditions, and we discuss our next steps for hypoxia biomarkers using an RNA-seq study., Summary: Gene expression biomarkers associated with salinity stress and thermal stress, but not hypoxic stress, exist in salmon gill tissue. Biomarkers were also associated with mortality.