Reduced lactate dehydrogenase activity in the heart and suppressed sex hormone levels are associated with female-biased mortality during thermal stress in Pacific salmon.

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, LDH), (iii) have 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 (CAT) activity]. 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.

Olfactory gene expression in migrating adult sockeye salmon Oncorhynchus nerka.

Expression of 12 olfactory genes was analysed in adult sockeye salmon Oncorhynchus nerka nearing spawning grounds and O. nerka that had strayed from their natal migration route. Variation was found in six of these genes, all of which were olfc olfactory receptors and had lower expression levels in salmon nearing spawning grounds. The results may reflect decreased sensitivity to natal water olfactory cues as these fish are no longer seeking the correct migratory route. The expression of olfactory genes during the olfactory-mediated spawning migration of Pacific salmon Oncorhynchus spp. is largely unexplored and these findings demonstrate a link between migratory behaviours and olfactory plasticity that provides a basis for future molecular research on salmon homing.

Maternal programming of offspring hypothalamic-pituitary-interrenal axis in wild sockeye salmon (Oncorhynchus nerka).

In fishes, maternal exposure to a stressor can influence offspring size and behavior. However, less is known about how maternal stress influences physiological processes in offspring, such as function of the hypothalamic-pituitary-interrenal (HPI) axis. We examined the impact of chronic maternal exposure to an acute chase stressor on the stress response/HPI activity of progeny in wild sockeye salmon (Oncorhynchus nerka). Resting plasma cortisol and brain preoptic area (POA) corticotropin-releasing factor (CRF) mRNA levels did not vary between offspring reared from undisturbed, control females and offspring reared from females exposed to the stressor. However, resting levels of POA glucocorticoid receptors (GR1 and GR2), and head kidney melanocortin 2 receptor (MC2R), steroidogenic acute regulatory protein (StAR), and cytochrome P450 side chain cleavage enzyme (P450scc) were elevated in offspring reared from stressor-exposed females. Offspring reared from stressor-exposed females had lower plasma cortisol levels 1-h after an acute chase stressor compared to cortisol levels in offspring reared from control females. In offspring reared from chased females, mRNA levels of genes associated with cortisol biosynthesis were reduced in the head kidney post-chase. In offspring reared from control females, mRNA levels in the head kidney did not vary pre- to post-chase. Together, the results of the present study suggest maternal programming of progeny with respect to baseline and stressor-induced mediators of HPI axis activity.

A survey of microparasites present in adult migrating Chinook salmon (Oncorhynchus tshawytscha) in south-western British Columbia determined by high-throughput quantitative polymerase chain reaction.

Microparasites play an important role in the demography, ecology and evolution of Pacific salmonids. As salmon stocks continue to decline and the impacts of global climate change on fish populations become apparent, a greater understanding of microparasites in wild salmon populations is warranted. We used high-throughput, quantitative PCR (HT-qRT-PCR) to rapidly screen 82 adult Chinook salmon from five geographically or genetically distinct groups (mostly returning to tributaries of the Fraser River) for 45 microparasite taxa. We detected 20 microparasite species, four of which have not previously been documented in Chinook salmon, and four of which have not been previously detected in any salmonids in the Fraser River. Comparisons of microparasite load to blood plasma variables revealed some positive associations between Flavobacterium psychrophilum, Cryptobia salmositica and Ceratonova shasta and physiological indices suggestive of morbidity. We include a comparison of our findings for each microparasite taxa with previous knowledge of its distribution in British Columbia.

Extensive feeding on sockeye salmon Oncorhynchus nerka smolts by bull trout Salvelinus confluentus during initial outmigration into a small, unregulated and inland British Columbia river.

Stomach contents were collected and analysed from 22 bull trout Salvelinus confluentus at the edge of the Chilko Lake and Chilko River in British Columbia, Canada, during spring outmigration of sockeye salmon Oncorhynchus nerka smolts. Twenty of the 22 (>90%) stomachs contained prey items, virtually all identifiable prey items were outmigrant O. nerka smolts and stomach contents represented a large portion (0·0-12·6%) of estimated S. confluentus mass. The results demonstrate nearly exclusive and intense feeding by S. confluentus on outmigrant smolts, and support recent telemetry observations of high disappearance rates of O. nerka smolts leaving large natural lake systems prior to entering high-order unregulated river systems.

Mother knows best, even when stressed? Effects of maternal exposure to a stressor on offspring performance at different life stages in a wild semelparous fish.

The environment mothers are exposed to has resonating effects on offspring performance. In iteroparous species, maternal exposure to stressors generally results in offspring ill-equipped for survival. Still, opportunities for future fecundity can offset low quality offspring. Little is known, however, as to how intergenerational effects of stress manifest in semelparous species with only a single breeding episode. Such mothers would suffer a total loss of fitness if offspring cannot survive past multiple life stages. We evaluated whether chronic exposure of female sockeye salmon (Oncorhynchus nerka) to a chase stressor impaired offspring performance traits. Egg size and early offspring survival were not influenced by maternal exposure to the repeated acute stressor. Later in development, fry reared from stressed mothers swam for shorter periods of time but possessed a superior capacity to re-initiate bouts of burst swimming. In contrast to iteroparous species, the mechanisms driving the observed effects do not appear to be related to cortisol, as egg hormone concentrations did not vary between stressed and undisturbed mothers. Sockeye salmon appear to possess buffering strategies that protect offspring from deleterious effects of maternal stress that would otherwise compromise progeny during highly vulnerable stages of development. Whether stressed sockeye salmon mothers endow offspring with traits that are matched or mismatched for survival in the unpredictable environment they encountered is discussed. This study highlights the importance of examining intergenerational effects among species-specific reproductive strategies, and across offspring life history to fully determine the scope of impact of maternal stress.

Using vitality indicators to predict survival of aquatic animals released from fisheries.

Estimating the survival probability of animals released from fisheries can improve the overall understanding of animal biology with implications for fisheries management, conservation and animal welfare. Vitality indicators are simple visual measures of animal condition that change in response to stressors (like fisheries capture) and can be assessed to predict post-release survival. These indicators typically include immediate reflex responses which are typically combined into a score. Vitality indicators are straight-forward and non-invasive metrics that allow users to quantify how close (or far) an animal is from a normal, 'healthy' or baseline state, which in turn can be correlated with outcomes such as survival probability, given appropriate calibration. The literature on using vitality indicators to predict post-release survival of animals has grown rapidly over the past decade. We identified 136 papers that used vitality indicators in a fisheries context. These studies were primarily focused on marine and freshwater fishes, with a few examples using herptiles and crustaceans. The types of vitality indicators are diverse and sometimes taxa-specific (e.g. pinching leg of turtles, spraying water at nictitating membrane of sharks) with the most commonly used indicators being those that assess escape response or righting response given the vulnerability of animals when those reflexes are impaired. By presenting Pacific salmon fisheries as a case study, we propose a framework for using vitality indicators to predict survival across taxa and fisheries.

Calibrating acoustic acceleration transmitters for estimating energy use by wild adult Pacific salmon.

This study is the first to calibrate acceleration transmitters with energy expenditure using a vertebrate model species. We quantified the relationship between acoustic accelerometer output and oxygen consumption across a range of swim speeds and water temperatures for Harrison River adult sockeye salmon (Oncorhynchus nerka). First, we verified that acceleration transmitters with a sampling frequency of 10 Hz could be used as a proxy for movement in sockeye salmon. Using a mixed effects model, we determined that tailbeat frequency and acceleration were positively correlated (p<0.0001), independent of tag ID. Acceleration (p<0.0001) was positively related to swim speed while fork length (p=0.051) was negatively related to swim speed. Oxygen consumption and accelerometer output (p<0.0001) had a positive linear relationship and were temperature dependent (p<0.0001). There were no differences in swim performance (F(2,12)=1.023, p=0.820) or oxygen consumption (F(1,12)=0.054, p=0.332) between tagged and untagged individuals. Five tagged fish were released into the Fraser River estuary and manually tracked. Of the five fish, three were successfully tracked for 1h. The above relationships were used to determine that the average swim speed was 1.25±0.03 body lengths s(-1) and cost of transport was 3.39±0.17 mg O(2) kg(-1)min(-1), averaged across the three detected fish. Acceleration transmitters can be effectively used to remotely evaluate fine-scale behavior and estimate energy consumption of adult Pacific salmon throughout their homeward spawning migration.

Provenance matters: thermal reaction norms for embryo survival among sockeye salmon Oncorhynchus nerka populations.

Differences in thermal tolerance during embryonic development in Fraser River sockeye salmon Oncorhynchus nerka were examined among nine populations in a controlled common-garden incubation experiment. Forcing embryonic development at an extreme temperature (relative to current values) of 16° C, representing a future climate change scenario, significantly reduced survival compared to the more ecologically moderate temperature of 10° C (55% v. 93%). Survival at 14° C was intermediate between the other two temperatures (85%). More importantly, this survival response varied by provenance within and between temperature treatments. Thermal reaction norms showed an interacting response of genotype and environment (temperature), suggesting that populations of O. nerka may have adapted differentially to elevated temperatures during incubation and early development. Moreover, populations that historically experience warmer incubation temperatures at early development displayed a higher tolerance for warm temperatures. In contrast, thermal tolerance does not appear to transcend life stages as adult migration temperatures were not related to embryo thermal tolerance. The intra-population variation implies potential for thermal tolerance at the species level. The differential inter-population variation in thermal tolerance that was observed suggests, however, limited adaptive potential to thermal shifts for some populations. This infers that the intergenerational effects of increasing water temperatures may affect populations differentially, and that such thermally mediated adaptive selection may drive population, and therefore species, persistence.

Does among-population variation in burst swimming performance of sockeye salmon Oncorhynchus nerka fry reflect early life migrations?

Using a fixed-speed test, burst swimming performance was found to vary among nine populations of emergent sockeye salmon Oncorhynchus nerka fry reared in a common-garden environment. No consistent relationship was, however, detected between difficulty of fry migration (upstream v. downstream) to rearing areas and total burst swimming duration or bursting rate.

Low cardiac and aerobic scope in a coastal population of sockeye salmon Oncorhynchus nerka with a short upriver migration.

This study showed that a coastal population (Harrison) of Fraser River sockeye salmon Oncorhynchus nerka had a lower aerobic and cardiac scope compared with interior populations with more challenging upriver spawning migrations, providing additional support to the idea that Fraser River O. nerka populations have adapted physiologically to their local migratory environment.

Parental identity influences progeny responses to incubation thermal stress in sockeye salmon Onchorhynchus nerka.

The influence of individual parentage on progeny responses to early developmental temperature stress was examined in a cross-fertilization experiment using sockeye salmon Oncorhynchus nerka. Differences in survival, hatch timing and size were examined among five paternally linked and five maternally linked offspring families (Weaver Creek population, British Columbia, Canada) incubated at 12, 14 and 16° C from just after fertilization to hatch. Mean embryonic survival was significantly lower at 14 and 16° C; however, offspring families had substantially different survival responses across the thermal gradient (crossing reaction norms). Within temperature treatments, substantial variation in embryonic survival, alevin mass, time-to-hatch and hatch duration were attributable to family identity; however, most traits were governed by significant temperature-family interactions. For embryonic survival, large differences between families at 16° C were due to both female and male spawner influence, whereas inter-family differences were obscured at 14° C (high intra-family variation), and minimal at 12° C (only maternal influence detected). Despite post-hatch rearing under a common cool thermal regime, persistent effects of both temperature and parentage were detected in alevin and 3 week-old fry. Collectively, these findings highlight the crucial role that parental influences on offspring may have in shaping future selection within salmonid populations exposed to elevated thermal regimes. An increased understanding of parental and temperature influences and their persistence in early development will be essential to developing a more comprehensive view of population spawning success and determining the adaptive capacity of O. nerka populations in the face of environmental change.

Quantitative methods for analysing cumulative effects on fish migration success: a review.

It is often recognized, but seldom addressed, that a quantitative assessment of the cumulative effects, both additive and non-additive, of multiple stressors on fish survival would provide a more realistic representation of the factors that influence fish migration. This review presents a compilation of analytical methods applied to a well-studied fish migration, a more general review of quantitative multivariable methods, and a synthesis on how to apply new analytical techniques in fish migration studies. A compilation of adult migration papers from Fraser River sockeye salmon Oncorhynchus nerka revealed a limited number of multivariable methods being applied and the sub-optimal reliance on univariable methods for multivariable problems. The literature review of fisheries science, general biology and medicine identified a large number of alternative methods for dealing with cumulative effects, with a limited number of techniques being used in fish migration studies. An evaluation of the different methods revealed that certain classes of multivariable analyses will probably prove useful in future assessments of cumulative effects on fish migration. This overview and evaluation of quantitative methods gathered from the disparate fields should serve as a primer for anyone seeking to quantify cumulative effects on fish migration survival.

Dead fish swimming: a review of research on the early migration and high premature mortality in adult Fraser River sockeye salmon Oncorhynchus nerka.

Adult sockeye salmon Oncorhynchus nerka destined for the Fraser River, British Columbia are some of the most economically important populations but changes in the timing of their homeward migration have led to management challenges and conservation concerns. After a directed migration from the open ocean to the coast, this group historically would mill just off shore for 3-6 weeks prior to migrating up the Fraser River. This milling behaviour changed abruptly in 1995 and thereafter, decreasing to only a few days in some years (termed early migration), with dramatic consequences that have necessitated risk-averse management strategies. Early migrating fish consistently suffer extremely high mortality (exceeding 90% in some years) during freshwater migration and on spawning grounds prior to spawning. This synthesis examines multidisciplinary, collaborative research aimed at understanding what triggers early migration, why it results in high mortality, and how fisheries managers can utilize these scientific results. Tissue analyses from thousands of O. nerka captured along their migration trajectory from ocean to spawning grounds, including hundreds that were tracked with biotelemetry, have revealed that early migrants are more reproductively advanced and ill-prepared for osmoregulatory transition upon their entry into fresh water. Gene array profiles indicate that many early migrants are also immunocompromised and stressed, carrying a genomic profile consistent with a viral infection. The causes of these physiological changes are still under investigation. Early migration brings O. nerka into the river when it is 3-6° C warmer than historical norms, which for some late-run populations approaches or exceeds their critical maxima leading to the collapse of metabolic and cardiac scope, and mortality. As peak spawning dates have not changed, the surviving early migrants tend to mill in warm lakes near to spawning areas. These results in the accumulation of many more thermal units and longer exposures to freshwater diseases and parasites compared to fish that delay freshwater entry by milling in the cool ocean environment. Experiments have confirmed that thermally driven processes are a primary cause of mortality for early-entry migrants. The Fraser River late-run O. nerka early migration phenomenon illustrates the complex links that exist between salmonid physiology, behaviour and environment and the pivotal role that water temperature can have on population-specific migration survival.

Corrigendum to: Sex-specific differences in swimming, aerobic metabolism and recovery from exercise in adult coho salmon (Oncorhynchus kisutch) across ecologically relevant temperatures.

[This corrects the article DOI: 10.1093/conphys/coab016.].

Fitness and behavioral correlates of pre-stress and stress-induced plasma cortisol titers in pink salmon (Oncorhynchus gorbuscha) upon arrival at spawning grounds.

Semelparous Pacific salmon (Onchorynchus spp.) serve as an excellent model for examining the relationships between life history, behavior and individual variation in glucocorticoid (GC) stress hormone levels because reproductive behaviors are highly variable between individuals and failure to reproduce results in zero fitness. Pink salmon (O. gorbuscha) were intercepted upon arrival at the spawning grounds across three time periods. Pre-stress and stress-induced plasma cortisol concentrations were assessed in relation to behavior, longevity and reproductive success. Results revealed differences between sexes and with arrival time. The study period marked a year of high reproductive success and only nine females (12% of sample) failed to spawn. Female pre-spawn mortalities were characterized by significantly elevated stress-induced cortisol concentrations and decreased longevity as well as pre-stress cortisol above the normal range in pink salmon from the study area. Interestingly, reproductive behaviors were only associated with pre-stress cortisol levels. For females, aggression and mate interaction time were reduced in individuals with elevated pre-stress cortisol concentrations. In males, a similar negative relationship between pre-stress cortisol concentration and mate interaction time was detected. The observed behavioral correlations are likely a factor of social status where dominant individuals, known to have higher reproductive success, are characterized by lower cortisol levels relative to subordinate conspecifics. Findings show both elevated pre-stress and stress-induced cortisol concentrations at arrival to the spawning grounds to be associated with reduced survival.

Temporal changes in blood variables during final maturation and senescence in male sockeye salmon Oncorhynchus nerka: reduced osmoregulatory ability can predict mortality.

This study is the first to characterize temporal changes in blood chemistry of individuals from one population of male sockeye salmon Oncorhynchus nerka during the final 6 weeks of sexual maturation and senescence in the freshwater stage of their spawning migration. Fish that died before the start of their historic mean spawning period (c. 5 November) were characterized by a 20-40% decrease in plasma osmolality, chloride and sodium, probably representing a complete loss of osmoregulatory ability. As fish became moribund, they were further characterized by elevated levels of plasma cortisol, lactate and potassium. Regressions between time to death and plasma chloride (8 October: P < 0·001; 15 October: P < 0·001) indicate that plasma chloride was a strong predictor of longevity in O. nerka. That major plasma ion levels started to decline 2-10 days (mean of 6 days) before fish became moribund, and before other stress, metabolic or reproductive hormone variables started to change, suggests that a dysfunctional osmoregulatory system may initiate rapid senescence and influence other physiological changes (i.e. elevated stress and collapsed reproductive hormones) which occur as O. nerka die on spawning grounds.

The efficacy of field techniques for obtaining and storing blood samples from fishes.

Prompted by the dramatic increase in the use of blood analyses in fisheries research and monitoring, this study investigated the efficacy of common field techniques for sampling and storing blood from fishes. Three questions were addressed: (1) Do blood samples taken via rapid caudal puncture (the 'grab-and-stab' technique) yield similar results for live v. sacrificed groups of fishes? (2) Do rapidly obtained caudal blood samples accurately represent blood properties of fishes prior to capture? (3) Does storage of whole blood in an ice slurry for a working day (8·5 h) modify the properties of the plasma? It was shown that haematocrit, plasma ions, metabolites, stress hormones and sex hormones of caudal blood samples were statistically similar when taken from live v. recently sacrificed groups of adult coho salmon Oncorhynchus kisutch. Moreover, this study confirmed by using paired blood samples from cannulated O. kisutch that blood acquired through the caudal puncture technique (mean ±s.e. 142 ± 26 s after capture) was representative of fish prior to capture. Long-term (8·5 h) cold storage of sockeye salmon Oncorhynchus nerka whole blood caused significant decreases in plasma potassium and chloride, and a significant increase in plasma glucose. Previous research has suggested that these changes largely result from net movements of ions and molecules between the plasma and erythrocytes, movements that can occur within minutes of storage. Thus, blood samples from fishes should be centrifuged as quickly as practicable in the field for separation of plasma and erythrocytes to prevent potentially misleading data.

Sex-specific differences in swimming, aerobic metabolism and recovery from exercise in adult coho salmon (Oncorhynchus kisutch) across ecologically relevant temperatures.

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 (U crit) 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.

Testing the synergistic effects of GnRH and testosterone on the reproductive physiology of pre-adult pink salmon Oncorhynchus gorbuscha.

To test the hypothesis that the hypothalmic gonadotropin-releasing hormone (GnRH) and testosterone (T) co-treatment stimulates both the hypothalmo-pituitary-gonadal (HPG) and hypothalmo-pituitary-interrenal axes, the reproductive and osmoregulatory responses of pre-adult pink salmon Oncorhynchus gorbuscha were compared after GnRH and T administration either alone or in combination. Relative to controls, neither GnRH nor T treatment resulted in significantly greater ovarian or testicular growth, but co-treatment significantly increased ovarian growth after 5 months. Interestingly, the stimulation was undetectable after 3 months. However, once daily photoperiod began shortening after the summer solstice, c. 2 months before the natural spawning date, GnRH+T-treated females were stimulated to produce larger ovaries. Final fish body length and the size of individual eggs did not differ among treatment groups. GnRH+T eggs, however, showed signs of advanced vitellogenesis relative to GnRH-treated and control eggs, whereas T-treated eggs became atretic. Testis size increased significantly from initial values and most males were spermiating, but this growth and development were independent of hormone treatments. Final plasma ion, metabolite and cortisol concentrations did not differ among treatment groups. It is concluded that GnRH+T co-treatment was effective in stimulating female but not male maturation. GnRH and T treatment, however, presumably had little effect on the hypothalmo-pituitary-interrenal axis as observed by ionoregulatory status.