Freshwater protected areas can preserve high-performance phenotypes in populations of a popular sportfish.

Recreational fishing has the potential to cause evolutionary change in fish populations; a phenomenon referred to as fisheries-induced evolution. However, detecting and quantifying the magnitude of recreational fisheries selection in the wild is inherently difficult, largely owing to the challenges associated with variation in environmental factors and, in most cases, the absence of pre-selection or baseline data against which comparisons can be made. However, exploration of recreational fisheries selection in wild populations may be possible in systems where fisheries exclusion zones exist. Lakes that possess intra-lake freshwater protected areas (FPAs) can provide investigative opportunities to evaluate the evolutionary impact(s) of differing fisheries management strategies within the same waterbody. To address this possibility, we evaluated how two physiological characteristics (metabolic phenotype and stress responsiveness) as well as a proxy for angling vulnerability, catch-per-unit-effort (CPUE), differed between populations of largemouth bass (Micropterus salmoides) inhabiting long-standing (>70 years active) intra-lake FPAs and adjacent, open access, main-lake areas. Fish from FPA populations had significantly higher aerobic scope (AS) capacity (13%) and CPUE rates compared with fish inhabiting the adjacent main-lake areas. These findings are consistent with theory and empirical evidence linking exploitation with reduced metabolic performance, supporting the hypothesis that recreational fishing may be altering the metabolic phenotype of wild fish populations. Reductions in AS are concerning because they suggest a reduced scope for carrying out essential life-history activities, which may result in fitness level implications. Furthermore, these results highlight the potential for unexploited FPA populations to serve as benchmarks to further investigate the evolutionary consequences of recreational fishing on wild fish and to preserve high-performance phenotypes.

Too stressed to eat: Investigating factors associated with appetite loss in subordinate rainbow trout.

Juvenile rainbow trout (Oncorhynchus mykiss) form dominance hierarchies in which subordinates experience chronic social stress and suppression of food intake. Here we tested the hypothesis that inhibition of food intake reflects increased expression of anorexigenic (appetite inhibiting) signals and decreased expression of orexigenic (appetite stimulating) signals. Trout were confined in pairs for 1 or 4 days, or were confined in pairs for 4 days and then allowed to recover from social interactions for 2 or 4 days; sham fish were handled identically but held alone. Subordinates did not feed during social interaction and had lower food intake than dominants or shams during recovery. In parallel, plasma cortisol (∼18-26x) and liver leptin (lep-a1) transcript abundance (∼10-14x) were elevated in subordinates during social interaction but not recovery, suggesting that these factors contributed to the suppression of food intake. Fish deemed likely to become subordinate based on inhibition of food intake in response to a mild stressor also showed elevated liver lep-a1 transcript abundance (∼5x). The moderate response in these fish coupled with a correlation between liver lep-a1 and cortisol suggest that stress-induced elevation of cortisol increased liver lep-a1 transcript abundance in subordinate trout, contributing to stress-induced suppression of food intake.

Role of cytosolic carbonic anhydrase Ca17a in cardiorespiratory responses to CO2 in developing zebrafish (Danio rerio).

The sensing of environmental fluctuations and initiation of appropriate physiological responses is crucial to homeostasis. Neuroepithelial cells (NECs) in fishes are putative chemoreceptors, resembling mammalian Type I (glomus) cells, that respond in vitro to changes in O2, CO2, NH3, and pH. Cytosolic carbonic anhydrase (Ca17a) is thought to be involved in CO2 sensing owing to its presence in NECs. Zebrafish (Danio rerio) lacking functional Ca17a were generated via CRISPR/Cas9 technology and used to assess the role of Ca17a in initiating the cardiorespiratory responses to elevated CO2 (hypercapnia). Unfortunately, the homozygous knockout mutants (ca17a-/-) did not survive more than ∼12-14 days postfertilization (dpf), restricting experiments to early developmental stages (4-8 dpf). Changes in ventilation (fV) and cardiac (fH) frequency in response to hypercapnia (1% CO2) in wild-type (ca17a+/+), heterozygous (ca17a+/-) and ca17a-/- fish were used to investigate Ca17a-dependent CO2 sensing and downstream signaling. Wild-type fish exhibited hyperventilation during hypercapnia as indicated by an increase in fV. In the ca17a-/- fish, the hyperventilatory response was attenuated markedly but only at 8 dpf. Hypercapnic tachycardia was observed for all genotypes and did not appear to be influenced by the absence of Ca17a. Interestingly, ca17a-/- fish exhibited a significantly lower resting fH that became more pronounced as the fish aged. The decrease in resting fH was prevented ("rescued") when ca17a-/- embryos were injected with ca17a mRNA. Collectively, the results of this study support a role for Ca17a in promoting hyperventilation during hypercapnia in larval zebrafish and suggest a previously unrecognized role for Ca17a in determining resting heart rate.

Early-life stress influences ion balance in developing zebrafish (Danio rerio).

As a key endocrine axis involved in responding to stress, the hypothalamic-pituitary-interrenal axis plays dual roles in mobilizing energy and maintaining ionic/osmotic balance in fishes. Although these roles have been examined independently in detail in adult fishes, less attention has been paid to the effects of an endogenous stress response during early life, particularly with respect to its potential effects on ionic/osmotic balance. The present study tested the hypothesis that exposure of zebrafish to stress during early development would alter ion balance later in life. Zebrafish at three developmental stages (4, 7, or 15 days post-fertilization, dpf) were subjected to an air-exposure stressor twice a day for 2 days, causing elevation of whole-body cortisol levels. Individuals stressed early in life exhibited decreased survival and growth, altered cortisol responses to a subsequent air-exposure stressor, and increased whole-body Na+ and Ca2+ concentrations. Changes in whole-body Ca2+ concentrations were accompanied by increased ionocyte abundance at 7 dpf and increased rates of Ca2+ uptake from the environment. Differences in whole-body ion concentrations at 15 and 35 dpf were not accompanied by altered ion uptake rates. Across all ages examined, air-exposure stress experienced at 7 dpf was particularly effective at eliciting phenotypic changes, suggesting a critical window at this age for a stress response to influence development. These findings demonstrate that early-life stress in zebrafish triggers developmental plasticity, with age-dependent effects on both the cortisol stress axis and ion balance.

The role of TASK-2 channels in CO2 sensing in zebrafish (Danio rerio).

Peripheral chemosensitivity in fishes is thought to be mediated by serotonin-enriched neuroepithelial cells (NECs) that are localized to the gills of adults and the integument of larvae. In adult zebrafish (Danio rerio), branchial NECs are presumed to mediate the cardiorespiratory reflexes associated with hypoxia or hypercapnia, whereas in larvae, there is indirect evidence linking cutaneous NECs to hypoxic hyperventilation and hypercapnic tachycardia. No study yet has examined the ventilatory response of larval zebrafish to hypercapnia, and regardless of developmental stage, the signaling pathways involved in CO2 sensing remain unclear. In the mouse, a background potassium channel (TASK-2) contributes to the sensitivity of chemoreceptor cells to CO2. Zebrafish possess two TASK-2 channel paralogs, TASK-2 and TASK-2b, encoded by kcnk5a and kcnk5b, respectively. The present study aimed to determine whether TASK-2 channels are expressed in NECs of larval zebrafish and whether they are involved in CO2 sensing. Using immunohistochemical approaches, TASK-2 protein was observed on the surface of NECs in larvae. Exposure of larvae to hypercapnia caused cardiac and breathing frequencies to increase, and these responses were blunted in fish experiencing TASK-2 and/or TASK-2b knockdown. The results of these experiments suggest that TASK-2 channels are involved in CO2 sensing by NECs and contribute to the initiation of reflex cardiorespiratory responses during exposure of larvae to hypercapnia.

Comparison of vegetable shortening and cocoa butter as vehicles for cortisol manipulation in Salmo trutta.

This study demonstrates that vegetable shortening and cocoa butter are two effective vehicles for intraperitoneal cortisol implants in juvenile teleosts, specifically brown trout Salmo trutta, residing in north temperate freshwater environments. Each vehicle showed a different pattern of cortisol elevation. Vegetable shortening was found to be a more suitable vehicle for long-term cortisol elevation [elevated at 3, 6 and 9 days post treatment (dpt)], while cocoa butter may be better suited for short-term cortisol elevation (only elevated at 3 dpt). Additionally, plasma cortisol levels were higher with cortisol-vegetable shortening than with cortisol-cocoa butter implants. Plasma glucose levels were elevated 6 and 9 dpt for fishes injected with cortisol-vegetable shortening, but did not change relative to controls and shams in cortisol-cocoa butter fishes. In conclusion, vegetable shortening and cocoa butter are both viable techniques for cortisol manipulation in fishes in temperate climates, providing researchers with different options depending on study objectives.

Regulation of energy metabolism during social interactions in rainbow trout: a role for AMP-activated protein kinase.

Rainbow trout (Oncorhynchus mykiss) confined in pairs form social hierarchies in which subordinate fish typically experience fasting and high circulating cortisol levels, resulting in low growth rates. The present study investigated the role of AMP-activated protein kinase (AMPK) in mediating metabolic adjustments associated with social status in rainbow trout. After 3 days of social interaction, liver AMPK activity was significantly higher in subordinate than dominant or sham (fish handled in the same fashion as paired fish but held individually) trout. Elevated liver AMPK activity in subordinate fish likely reflected a significantly higher ratio of phosphorylated AMPK (phospho-AMPK) to total AMPK protein, which was accompanied by significantly higher AMPKα1 relative mRNA abundance. Liver ATP and creatine phosphate concentrations in subordinate fish also were elevated, perhaps as a result of AMPK activity. Sham fish that were fasted for 3 days exhibited effects parallel to those of subordinate fish, suggesting that low food intake was an important trigger of elevated AMPK activity in subordinate fish. Effects on white muscle appeared to be influenced by the physical activity associated with social interaction. Overall, muscle AMPK activity was significantly higher in dominant and subordinate than sham fish. The ratio of phospho-AMPK to total AMPK protein in muscle was highest in subordinate fish, while muscle AMPKα1 relative mRNA abundance was elevated by social dominance. Muscle ATP and creatine phosphate concentrations were high in dominant and subordinate fish at 6 h of interaction and decreased significantly thereafter. Collectively, the findings of the present study support a role for AMPK in mediating liver and white muscle metabolic adjustments associated with social hierarchy formation in rainbow trout.

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.

Stress, nutrition and parental care in a teleost fish: exploring mechanisms with supplemental feeding and cortisol manipulation.

Parental care is an essential life-history component of reproduction for many animal species, and it entails a suite of behavioural and physiological investments to enhance offspring survival. These investments can incur costs to the parent, reducing their energetic and physiological condition, future reproductive capabilities and survival. In fishes, relatively few studies have focused on how these physiological costs are mediated. Male smallmouth bass provide parental care for developing offspring until the brood reaches independence. During this energetically demanding life stage, males cease active foraging as they vigorously defend their offspring. Experimental manipulation of cortisol levels (via implantation) and food (via supplemental feeding) in parental males was used to investigate the fitness consequences of parental care. Improving the nutritional condition of nest-guarding males increased their reproductive success by reducing premature nest abandonment. However, supplemental feeding and cortisol treatment had no effect on parental care behaviours. Cortisol treatment reduced plasma lymphocyte numbers, but increased neutrophil and monocyte concentrations, indicating a shift in immune function. Supplemental feeding improved the physiological condition of parental fish by reducing the accumulation of oxidative injury. Specifically, supplemental feeding reduced the formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG) on DNA nucleotides. Increasing the nutritional condition of parental fish can reduce the physiological cost associated with intensive parental activity and improve overall reproductive success, illustrating the importance of nutritional condition as a key modulator of parental fitness.

Regulation of hypothalamic-pituitary-interrenal axis function in male smallmouth bass (Micropterus dolomieu) during parental care.

Male smallmouth bass (Micropterus dolomieu) provide sole parental care until offspring reach independence, a period of several weeks. During the early parental care period when males are guarding fresh eggs (MG-FE), cortisol responsiveness is attenuated; the response is re-established when males reach the end of the parental care period and are guarding free-swimming fry (MG-FSF). It was hypothesized that attenuation of the cortisol response in male smallmouth bass during early parental care reflected modulation of hypothalamic-pituitary-interrenal (HPI) axis function. Male smallmouth bass were sampled at the beginning (MG-FE) and end of the parental care period (MG-FSF), before and/or 25 min after exposure to a standardized stressor consisting of 3 min of air exposure. Repeated sampling of stressed fish for analysis of plasma cortisol and adrenocorticotropic hormone (ACTH) levels was carried out. Males significantly elevated both plasma cortisol and ACTH levels when guarding free-swimming fry but not during early parental care. Control and stressed fish were terminally sampled for tissue mRNA abundance of preoptic area (POA) and hypothalamic corticotropin-releasing factor (CRF) as well as head kidney melanocortin 2 receptor (MC2R), steroidogenic acute regulatory protein (StAR) and cytochrome P450 side chain cleavage enzyme (P450scc). No significant differences in either hypothalamus CRF or head kidney P450scc mRNA abundance were found across parental care stages or in response to stress. However, POA CRF mRNA abundance and interrenal cell MC2R and StAR mRNA abundances failed to increase in response to stress in MG-FE. Thus, the attenuated cortisol response in males guarding fresh eggs may be explained by hypoactive HPI axis function in response to stress. The present is one of few studies, and the first teleost study, to address the mechanisms underlying resistance to stress during the reproductive/parental care period.

Social stress modulates the cortisol response to an acute stressor in rainbow trout (Oncorhynchus mykiss).

In rainbow trout (Oncorhynchus mykiss) of subordinate social status, circulating cortisol concentrations were elevated under resting conditions but the plasma cortisol and glucose responses to an acute stressor (confinement in a net) were attenuated relative to those of dominant trout. An in vitro head kidney preparation, and analysis of the expression of key genes in the stress axis prior to and following confinement in a net were then used to examine the mechanisms underlying suppression of the acute cortisol stress response in trout experiencing chronic social stress. With porcine adrenocorticotropic hormone (ACTH) as the secretagogue, ACTH-stimulated cortisol production was significantly lower for head kidney preparations from subordinate trout than for those from dominant trout. Dominant and subordinate fish did not, however, differ in the relative mRNA abundance of melanocortin-2 receptor (MC2R), steroidogenic acute regulatory protein (StAR) or cytochrome P450 side chain cleavage enzyme (P450scc) within the head kidney, although the relative mRNA abundance of these genes was significantly higher in both dominant and subordinate fish than in sham trout (trout that did not experience social interactions but were otherwise treated identically to the dominant and subordinate fish). The relative mRNA abundance of all three genes was significantly higher in trout exposed to an acute net stressor than under control conditions. Upstream of cortisol production in the stress axis, plasma ACTH concentrations were not affected by social stress, nor was the relative mRNA abundance of the binding protein for corticotropin releasing factor (CRF-BP). The relative mRNA abundance of CRF in the pre-optic area of subordinate fish was significantly higher than that of dominant or sham fish 1h after exposure to the stressor. Collectively, the results indicate that chronic social stress modulates cortisol production at the level of the interrenal cells, resulting in an attenuated cortisol response to an acute stressor.

Fish in hot water: hypoxaemia does not trigger catecholamine mobilization during heat shock in rainbow trout (Oncorhynchus mykiss).

Rainbow trout (Oncorhynchus mykiss) exposed to an acute heat shock (1 h at 25 °C after raising water temperature from 13 °C to 25 °C over 4 h) mount a significant catecholamine response. The present study investigated the proximate mechanisms underlying catecholamine mobilization. Trout exposed to heat shock in vivo exhibited a significant reduction in arterial O(2) tension, but arterial O(2) concentration was not affected by heat shock, nor was catecholamine release during heat shock prevented by prior and concomitant exposure to hyperoxia (to prevent the fall in arterial O(2) tension). Thus, catecholamine mobilization probably was not triggered by impaired blood O(2) transport. Heat-shocked trout also exhibited an elevation of arterial CO(2) tension coupled with a fall in arterial pH, but these factors are not expected to trigger catecholamine release. The changes in blood O(2) and CO(2) tension occurred despite a significant hyperventilatory response to heat shock. Future studies should investigate whether catecholamine mobilization during heat shock in rainbow trout is triggered by a specific effect of high temperature activating the sympathetic nervous system via a thermosensitive transient receptor potential channel.

Intracellular carbonic anhydrase contributes to the red blood cell adrenergic response in rainbow trout Oncorhynchus mykiss.

In many teleost fish, catecholamines activate a red blood cell (RBC) Na+/H+ exchanger (ßNHE), raising RBC intracellular pH to protect haemoglobin-O2 loading. The present study tested the hypothesis that RBC intracellular carbonic anhydrase (CA) contributes to this adrenergic response. The pH of rainbow trout (Oncorhynchus mykiss) blood was monitored continuously in vitro using blood flowing in a semi-closed loop or in vivo using an extracorporeal circulation. Addition or injection of isoproterenol activated the ßNHE, causing blood pH to fall (in vitro ΔpH = -0.28±0.03 pH units, N=16; in vivo, -0.12±0.02 pH units, N=6). Both in vitro and in vivo, inhibition of RBC CA by acetazolamide significantly decreased the magnitude of the adrenergic response (in vitro, ΔpH=-0.22±0.02 pH units, N=16; in vivo, -0.02±0.01 pH units, N=6) as well as the rate of recovery of blood pH following the adrenergic response. These results support the hypothesis that RBC intracellular CA plays an important role in the RBC adrenergic response of rainbow trout, and fuel speculation that interspecific differences in RBC CA activity are associated with the magnitude of the RBC adrenergic response.

Acid-base balance during social interactions in rainbow trout (Oncorhynchus mykiss).

Socially subordinate rainbow trout (Oncorhynchus mykiss) experience chronic stress that impacts upon a variety of physiological functions, including Na(+) regulation. Owing to the tight coupling between Na(+) and Cl(-) uptake and, respectively, H(+) and HCO(3)(-) loss at the gill, ionoregulatory changes associated with social status may affect acid-base regulation. The present study assessed the responses of dominant, subordinate and control trout to hypercapnia (1% CO(2)) to test this hypothesis. Social status appeared to impact net acid excretion (J(net)H(+)) as subordinate individuals failed to increase net acid flux in response to hypercapnia. However, blood acid-base status was found to be unaffected by social status before or during hypercapnic exposure, indicating that subordinate fish were as effective as dominant or control trout in achieving compensation for the acid-base disturbance induced by hypercapnic exposure. Compensation in all groups involved decreasing Cl(-) uptake in response to hypercapnia. The branchial activities of both Na(+),K(+)-ATPase (NKA) and V-type H(+)-ATPase were affected by social interactions and/or exposure to hypercapnia. Branchial NKA activity was higher but V-ATPase activity was lower in control fish than in dominant or subordinate trout. In addition, control and subordinate but not dominant trout exposed to 24h of hypercapnia exhibited significantly higher branchial V-ATPase activity than fish maintained in normocapnia. Collectively, the data suggest that subordinate trout are able to regulate blood pH during a respiratory acidosis.

Low social status impairs hypoxia tolerance in rainbow trout (Oncorhynchus mykiss).

In the present study, chronic behavioural stress resulting from low social status affected the physiological responses of rainbow trout (Oncorhynchus mykiss) to a subsequent acute stressor, exposure to hypoxia. Rainbow trout were confined in fork-length matched pairs for 48-72 h, and social rank was assigned based on behaviour. Dominant and subordinate fish were then exposed individually to graded hypoxia (final water PO(2), PwO(2) = 40 Torr). Catecholamine mobilization profiles differed between dominant and subordinate fish. Whereas dominant fish exhibited generally low circulating catecholamine levels until a distinct threshold for release was reached (PwO(2) = 51.5 Torr corresponding to arterial PO(2), PaO(2) = 24.1 Torr), plasma catecholamine concentrations in subordinate fish were more variable and identification of a distinct threshold for release was problematic. Among fish that mobilized catecholamines (i.e. circulating catecholamines rose above the 95% confidence interval around the baseline value), however, the circulating levels achieved in subordinate fish were significantly higher (459.9 ± 142.2 nmol L(-1), mean ± SEM, N = 12) than those in dominant fish (130.9 ± 37.9 nmol L(-1), N = 12). The differences in catecholamine mobilization occurred despite similar P(50) values in dominant (22.0 ± 1.5 Torr, N = 6) and subordinate (22.1 ± 2.2 Torr, N = 8) fish, and higher PaO(2) values in subordinate fish under severely hypoxic conditions (i.e. PwO(2) < 60 Torr). The higher PaO(2) values of subordinate fish likely reflected the greater ventilatory rates and amplitudes exhibited by these fish during severe hypoxia. At the most severe level of hypoxia, subordinate fish were unable to defend arterial blood O(2) content, which fell to approximately half (0.60 ± 0.13 mL O(2) g(-1) haemoglobin, N = 9) that of dominant fish (1.08 ± 0.09 mL O(2) g(-1) haemoglobin, N = 9). Collectively, these data indicate that chronic social stress impacts the ability of trout to respond to the additional, acute stress of hypoxia.

Condition dependent intra-individual repeatability of stress-induced cortisol in a freshwater fish.

The glucocorticoid (GC) stress response is thought to be an individual trait associated with behaviour and life history strategies. Studies exploring such relationships typically assume measured hormone values to be repeatable within an individual. However, repeatability of GCs has proven variable in wild animals and underlying reasons remain unknown. We assessed individual repeatability of circulating stress-induced cortisol, the primary GC in teleost fish, and glucose concentrations in a wild teleost fish held under consistent laboratory conditions. We also tested the hypothesis that the magnitude of intra-individual variability in stress-induced cortisol concentrations ("cortisol variability") is influenced by body condition. Wild-caught bluegill sunfish (Lepomis macrochirus) were subjected to repeated standardized stressors and blood sampled (3 times over 6 days) once cortisol concentrations peaked. Various indicators of fish condition, both whole body and physiological, were also measured. Overall, stress-induced circulating cortisol concentrations were repeatable but stress-induced glucose was not. Cortisol variability was related to Fulton's condition factor and size (eviscerated mass) where smaller fish in poor condition exhibited increased cortisol variability. The findings have implications for the interpretation of studies that examine correlates of GC concentrations as they suggest consistency in stress responsiveness is influenced by factors such as size and condition.

Compensatory regulation of acid-base balance during salinity transfer in rainbow trout (Oncorhynchus mykiss).

In seawater-acclimated rainbow trout (Oncorhynchus mykiss), base secretion into the intestine is a key component of the intestinal water absorption that offsets osmotic water loss to the marine environment. Acid-base balance is maintained by the matched excretion of acid equivalents via other routes, presumably the gill and/or kidney. The goal of the present study was to examine acid-base balance in rainbow trout upon transfer to more dilute environments, conditions under which base excretion into the intestine is predicted to fall, requiring compensatory adjustments of acid excretion at the gill and/or kidney if acid-base balance is to be maintained. Net acid excretion via the gill/kidney and rectal fluid, and blood acid-base status were monitored in seawater-acclimated rainbow trout maintained in seawater or transferred to iso-osmotic conditions. As predicted, transfer to iso-osmotic conditions significantly reduced base excretion into the rectal fluid (by ~48%). Transfer to iso-osmotic conditions also significantly reduced the excretion of titratable acidity via extra-intestinal routes from 183.4 ± 71.3 to -217.5 ± 42.7 µmol kg(-1) h(-1) (N = 7). At the same time, however, ammonia excretion increased significantly during iso-osmotic transfer (by ~72%) so that the apparent overall reduction in net acid excretion (from 419.7 ± 92.9 to 189.2 ± 76.5 µmol kg(-1 )h(-1); N = 7) was not significant. Trout maintained blood acid-base status during iso-osmotic transfer, although arterial pH was significantly higher in transferred fish than in those maintained in seawater. To explore the mechanisms underlying these adjustments of acid-base regulation, the relative mRNA expression and where possible, activity of a suite of proteins involved in acid-base balance were examined in intestine, gill and kidney. At the kidney, reduced mRNA expression of carbonic anhydrase (CA; cytosolic and membrane-associated CA IV), V-type H(+)-ATPase, and Na(+)/HCO(3) (-) co-transporter were consistent with a reduced role in net acid excretion following iso-osmotic transfer. Changes in relative mRNA expression and/or activity at the intestine and gill were consistent with the roles of these organs in osmotic rather than acid-base regulation. Overall, the data emphasize the coordination of acid-base, osmoregulatory and ionoregulatory processes that occur with salinity transfer in a euryhaline fish.

The glucocorticoid stress response is repeatable between years in a wild teleost fish.

Patterns of glucocorticoid (GC) hormone regulation exhibit considerable inter-individual variation that is often examined relative to individual traits and fitness measures. Although stress-induced GC concentrations are repeatable within an individual in captive populations, this assumption remains untested in wild animals in their natural environment across longer time periods. We assessed the repeatability of baseline and post-stress GC concentrations in a wild teleost fish. Largemouth bass (Micropterus salmoides) were captured and subjected to a standard stress protocol and then stocked into a small research lake. Upon recapture by angling up to 1 year later (n = 26), fish were re-sampled following identical methods. After controlling for a strong effect of water temperature, we confirmed repeatability of post-stress cortisol concentrations despite stress presumed to accompany relocation. We documented no consistency in baseline GC concentrations. This study serves as an important validation for the use of post-stress cortisol concentrations as an individual trait. However, the effect size of repeatability was lower than that found in other taxa. Results also bring forth the reality that environmental variables such as temperature must be considered in studies where these factors can vary, such as when sampling wild animals at liberty.