Egg Cortisol Research Articles

Mimicking Transgenerational Signals of Future Stress: Thermal Tolerance of Juvenile Chinook Salmon Is More Sensitive to Elevated Rearing Temperature Than Exogenously Increased Egg Cortisol

Elevated temperatures resulting from climate change are expected to disproportionately affect ectotherms given their biological function has a direct link to environmental temperature. Thus, as climate change leads to rapid increases in water temperatures in rivers, aquatic ectotherms such as fish may be highly impacted. Organisms can respond to these stressors through flexible and rapid phenotypic change induced via developmental and/or transgenerational plasticity. In oviparous species, gravid females may translate environmental stress across generations via increased exposure of eggs to maternally-derived glucocorticoids (i.e., maternal stress), which has been shown to result in diverse phenotypic effects in offspring. Recent studies suggest these phenotypic changes from maternal glucocorticoids may elicit predictive adaptive responses, where offspring exposed to maternal stress may be better prepared for the stressful environment they will encounter (i.e., environmental match hypothesis). We applied the environmental match hypothesis to examine whether a prenatal exogenous increase in egg cortisol may prepare Chinook salmon offspring (Oncorhynchus tshawytscha) to cope with thermal challenges after being reared in chronically elevated temperatures. Specifically, we exposed eggs to aqueous bath of cortisol-dosed (1000ng/mL) or control (0ng/ml) solutions, and then raised both treatments at current (+0°C—contemporary ambient river temperature) or elevated (+3°C—projected future river temperature) thermal regimes. We quantified thermal performance in fish 7-9 month post fertilization using two methods: via critical thermal maximum (CTMax), and energetic responses (in plasma cortisol, glucose, and lactate) to environmentally relevant, but challenging thermal spikes over 3 days. Overall, we found that exposure to elevated rearing temperatures had a large impact on thermal tolerance, where elevated-temperature reared offspring had significantly higher CTMax. In comparison, egg cortisol treatment had little to no clear effects on CTMax and blood energetic response. Our study demonstrates the importance of elevated water temperatures as an inducer of offspring phenotypes (via early developmental cues), and highlights the significance of examining offspring performance in environments with ecologically relevant stressors.

Exposure to exogenous egg cortisol does not rescue juvenile Chinook salmon body size, condition, or survival from the effects of elevated water temperatures.

Climate change is leading to altered temperature regimes which are impacting aquatic life, particularly for ectothermic fish. The impacts of environmental stress can be translated across generations through maternally derived glucocorticoids, leading to altered offspring phenotypes. Although these maternal stress effects are often considered negative, recent studies suggest this maternal stress signal may prepare offspring for a similarly stressful environment (environmental match). We applied the environmental match hypothesis to examine whether a prenatal stress signal can dampen the effects of elevated water temperatures on body size, condition, and survival during early development in Chinook salmon Oncorhynchus tshawytscha from Lake Ontario, Canada. We exposed fertilized eggs to prenatal exogenous egg cortisol (1,000 ng/ml cortisol or 0 ng/ml control) and then reared these dosed groups at temperatures indicative of current (+0°C) and future (+3°C) temperature conditions. Offspring reared in elevated temperatures were smaller and had a lower survival at the hatchling developmental stage. Overall, we found that our exogenous cortisol dose did not dampen effects of elevated rearing temperatures (environmental match) on body size or early survival. Instead, our eyed stage survival indicates that our prenatal cortisol dose may be detrimental, as cortisol‐dosed offspring raised in elevated temperatures had lower survival than cortisol‐dosed and control reared in current temperatures. Our results suggest that a maternal stress signal may not be able to ameliorate the effects of thermal stress during early development. However, we highlight the importance of interpreting the fitness impacts of maternal stress within an environmentally relevant context.

Interaction of egg cortisol and offspring experience influences stress-related behaviour and physiology in lake sturgeon

Quantifying transgenerational effects of stress is important to predict outcomes of anthropogenic disturbances for wildlife species. Maternal stress can programme physiological and behavioural phenotypes in offspring, which may be maladaptive if maternal and offspring environments are mismatched. We investigated effects of a match and mismatch between egg cortisol and offspring stress levels in lake sturgeon, Acipenser fulvescens, using artificially elevated egg cortisol levels (simulating maternal stress) and a chronic unpredictable stress regime for offspring after hatch. Offspring cortisol levels were quantified at baseline and after an acute stressor. Multiple measures of offspring swimming activity were assessed in behaviour trials. Individuals that experienced elevated egg cortisol and high offspring stress exhibited a diminished cortisol response to an acute stressor, but responses varied among offspring from different families. Results suggest that the interaction between maternal and offspring experience may cue an offspring phenotype that is adaptive in high-stress conditions. Principal components analysis characterizing interindividual variation in offspring behavioural variables showed that treatment significantly affected multivariate offspring response along the PC1 axis (associated with inactivity), and both treatment and family significantly affected response along the PC2 axis (associated with shorter distance moved). The largest differences for PC1 occurred between the ‘mismatch’ treatments (high egg cortisol and low offspring stress exhibiting lower activity; low egg cortisol and high offspring stress exhibiting higher activity), indicating that the combination of egg cortisol and offspring stress is more important in determining offspring behaviour than is egg cortisol or offspring stress alone. Findings suggest that family effects, such as genetic components or maternal experience, may mediate how the interaction of maternal and offspring stress influences offspring physiological and behavioural outcomes, and indicate the need for further research into environmental factors experienced by females that influence how offspring respond to egg cortisol and early life stress.

Glucocorticoids in Fish Eggs: Variation, Interactions with the Environment, and the Potential to Shape Offspring Fitness.

Wild and captive vertebrates face multiple stressors that all have the potential to induce chronic maternal stress (i.e., sustained, elevated plasma glucocorticoids), resulting in embryo exposure to elevated maternally derived glucocorticoids. In oviparous taxa such as fish, maternally derived glucocorticoids in eggs are known for their capacity to shape offspring phenotype. Using a variety of methodologies, scientists have quantified maternally derived levels of egg cortisol, the primary glucocorticoid in fishes, and examined the cascading effects of egg cortisol on progeny phenotype. Here we summarize and interpret the current state of knowledge on egg cortisol in fishes and the relationships linking maternal stress/state to egg cortisol and offspring phenotype/fitness. Considerable variation in levels of egg cortisol exists across species and among females within a species; this variation is hypothesized to be due to interspecific differences in reproductive life history and intraspecific differences in female condition. Outcomes of experimental studies manipulating egg cortisol vary both inter- and intraspecifically. Moreover, while exogenous elevation of egg cortisol (as a proxy for maternal stress) induces phenotypic changes commonly considered to be maladaptive (e.g., smaller offspring size), emerging work in other taxa suggests that there can be positive effects on fitness when the offspring's environment is taken into account. Investigations into (i) mechanisms by which egg cortisol elicits phenotypic change in offspring (e.g., epigenetics), (ii) maternal and offspring buffering capacity of cortisol, and (iii) factors driving natural variation in egg cortisol and how this variation affects offspring phenotype and fitness are all germane to discussions on egg glucocorticoids as signals of maternal stress.

Prenatal Stress Exposure Generates Higher Early Survival and Smaller Size without Impacting Developmental Rate in a Pacific Salmon.

Prenatal exposure to elevated glucocorticoids can act as a signal of environmental stress, resulting in modifications to offspring phenotype. While "negative" phenotypic effects (i.e., smaller size, slower growth) are often reported, recent research coupling phenotype with other fitness-related traits has suggested positive impacts of prenatal stress. Using captive Chinook salmon (Oncorhynchus tshawytscha), we treated eggs with biologically relevant cortisol levels-low (300 ng mL-1 ), high (1,000 ng mL-1 ), or control (0 ng mL-1 )-to examine the early-life impacts of maternally transferred stress hormones on offspring. Specifically, we measured early survival, rate of development, and multiple measures of morphology. Low and high cortisol dosing of eggs resulted in significantly higher survival compared to controls (37% and 24% higher, respectively). Fish reared from high dose eggs were structurally smaller compared to control fish, but despite this variation in structural size, exposure to elevated cortisol did not impact developmental rate. These results demonstrate that elevations in egg cortisol can positively influence offspring fitness through an increase in early survival while also altering phenotype at a critical life-history stage. Overall, these results suggest that exposure to prenatal stress may not always produce apparently negative impacts on offspring fitness and further proposes that complex phenotypic responses should be examined in relevant environmental conditions.

Determination of cortisol in lake sturgeon (Acipenser fulvescens) eggs by liquid chromatography tandem mass spectrometry.

Quantifying cortisol concentrations in fish eggs is important to understand the effects of environmental conditions on maternal physiological condition and on egg provisioning and quality. Data are particularly relevant to studies of the ecology of threatened species such as lake sturgeon (Aciperser fulvescens) as well as assessments of larval physical and behavioral phenotypes, fish health and caviar quality in sturgeon aquaculture. This study focuses on development of bioanalytical methods for high sensitivity and robust determination of cortisol in sturgeon eggs. Sample preparation was optimized after investigating protein precipitation and liquid-liquid extraction techniques. Ethyl acetate was found to be the most efficient solvent (recovery parameter) and also provided the best sample clean up (matrix effect parameter). The method was determined to be linear for cortisol concentrations between 0.025 and 100ng/mL. The limits of detection and quantification were 0.025 and 0.1ng/mL respectively. Intra- and inter-day performances of the method were validated at three concentrations (0.25; 10 and 100ng/mL). The method was applied to field-collected samples for the determination of endogenous cortisol in lake sturgeon eggs. Cortisol was detected in all egg samples and statistical analysis showed significant differences between fertilized and non-fertilized eggs.

Effects of experimentally elevated egg cortisol on offspring traits in two species of wild Pacific salmon

In fishes, elevated levels of cortisol in eggs can have carry-over effects on phenotypic and performance traits early in life. How responses to elevations in egg cortisol differ among species remains poorly understood. Using wild populations of chum salmon (Oncorhynchus keta) and sockeye salmon (O. nerka), we investigated whether experimentally-elevated concentrations of cortisol in newly fertilized eggs had effects on offspring morphology and/or burst swimming capacity. Immediately following fertilization, eggs were incubated for 2 h with water dosed with 0 ng/mL or 1000 ng/mL of cortisol. Embryos were reared to the fry life stage (complete yolk sac absorption). Morphology and burst swimming performance of fry were then assessed. Sockeye salmon fry reared from cortisol-treated eggs were smaller overall (i.e., smaller body, fins and eyes) compared to conspecifics reared from untreated eggs. In contrast, the morphology of chum salmon fry was not affected by the experimental elevation of egg cortisol. In both species, burst swimming duration was unaffected by egg cortisol treatment, while offspring reared from the cortisol-treated eggs initiated fewer bouts of burst swimming. Our results demonstrate that closely-related species can respond differently to elevations in egg cortisol, and not all offspring traits may be affected by these elevations in cortisol. Further efforts to establish links among offspring quality, maternal stress, and egg composition need to consider the potential for divergent responses among species and examine multiple measures of phenotype and performance throughout development.

Examining the relationships between egg cortisol and oxidative stress in developing wild sockeye salmon (Oncorhynchus nerka)

Maternally-derived hormones in oocytes, such as glucocorticoids (GCs), play a crucial role in embryo development in oviparous taxa. In fishes, maternal stressor exposure increases circulating and egg cortisol levels, the primary GC in fishes, as well as induces oxidative stress. Elevated egg cortisol levels modify offspring traits but whether maternal oxidative stress correlates with circulating and egg cortisol levels, and whether maternal/egg cortisol levels correlate with offspring oxidative stress have yet to be determined. The objective of this study was to examine the relationships among maternal and egg cortisol, and maternal and offspring oxidative stress to provide insight into the potential intergenerational effects of stressor exposure in sockeye salmon (Oncorhynchus nerka). Antioxidant concentration and oxidative stress were measured in maternal tissues (plasma, brain, heart and liver) as well as offspring developmental stages (pre-fertilization, 24h post-fertilization, eyed, and hatch), and were compared to both naturally-occurring and experimentally-elevated (via cortisol egg bath) levels of cortisol in eggs. Oxygen radical absorptive capacity of tissues from maternal sockeye salmon was measured spectrophotometrically and was not correlated with maternal or egg cortisol concentrations. Also, naturally-occurring and experimentally-elevated cortisol levels in eggs (to mimic maternal stress) did not affect oxidative stress or antioxidant capacity of the offspring. We conclude that the metrics of maternal stress examined in sockeye salmon (i.e., maternal/egg cortisol, maternal oxidative stress) are independent of each other, and that egg cortisol content does not influence offspring oxidative stress.

Effects of maternal stress and cortisol exposure at the egg stage on learning, boldness and neophobia in brook trout

The environment experienced by females can have long-lasting effects on offspring phenotype. The objective of this study was to determine if maternal stress-induced behaviour reprogramming in offspring is found in brook char and to test whether cortisol is the main mediator, by separating the potential effects of cortisol from that of other potential maternal factors. We exposed female brook trout (Salvelinus fontinalis) to different parallel treatments during the oogenesis period: undisturbed as controls (1) fed cortisol through food (2) or physically stressed by handling once a week (3). Additionally, we exposed half of the control eggs to a cortisol suspension before fertilisation (4). Cortisol consumption and handling did not elevate either maternal plasma or egg cortisol, although egg cortisol level was significantly increased when eggs were bathed in the suspension. We measured spatial learning and memory, boldness and neophobia in 6 month-old offspring and found no effects of treatments on learning, memory or behaviour. Our results suggest that the relationship between maternal stress, circulating and egg cortisol levels, other maternal factors, and behavioural reprogramming is context and species-specific.

Egg Cortisol Exposure Enhances Fearfulness in Larvae and Juvenile Rainbow Trout

AbstractWe investigated the effects of an early boost of cortisol exposure in rainbow trout (Oncorhynchus mykiss) eggs during fertilisation on subsequent behavioural responses when exposed to a sudden stimulus in larvae and juveniles. At 55 d post‐fertilisation (dpf), treatment had no effect on high accelerations occurring after a sudden event. At 146 dpf, these high accelerations were more frequent in cortisol‐treated fish than in controls. At 146 dpf also, swimming activity was increased in cortisol‐treated fish both before and after the sudden stimulus. This study underlines the important behavioural modifications in both larvae and juveniles, linked to a change in the surrounding environment of the embryo. Indeed, fish exposed to cortisol as eggs showed a higher level of fearfulness later in life. Our findings are of major interest for stress management in an aquaculture context and also allow for a better understanding of the long‐lasting effects of a permanent and/or acute stress – mediated by cortisol – that could be encountered by females, affecting population's life history trajectory.

Stress response to anthropogenic noise in Atlantic cod Gadus morhua L.

The potential effects of anthropogenic noise on the physiology of Atlantic cod have not been well described. The aim of the present study was to investigate the impact of anthropogenic noise on Atlantic cod stress response using cortisol as a biomarker as well as on broodstock spawning performance. Results showed that artificial noise consisting of a linear sweep from 100 to 1000Hz can induce a transient and mild cortisol elevation with a clear noise intensity dose response. In all cases plasma levels returned to baseline levels <1h post sound exposure. Daily exposure to a similar intensity and frequency noise range applied habitually to a broodstock population during the spawning window resulted in a significant reduction in total egg production and fertilisation rates thus reducing the total production of viable embryos by over 50%. In addition, a significant negative correlation between egg cortisol content and fertilisation rate was observed. These results confirm that cod can perceive noise generated within a frequency range of 100–1000Hz and display a heightened cortisol plasma level. In addition, anthropogenic noise can have negative impacts on cod spawning performances.

Egg cortisol treatment affects the behavioural response of coho salmon to a conspecific intruder and threat of predation

Knowledge continues to emerge on how maternally derived egg hormones influence progeny behaviour in oviparous taxa. Of particular interest is how glucocorticoids (GCs) in eggs, as a signal of maternal stress, affect offspring behavioural traits. To date, research has focused on effects of egg GCs on offspring behaviours within a single experimental context. Little is known regarding how egg GCs affect relationships among behaviours and across ecological contexts (i.e. behavioural syndromes). We explored how exogenously increased egg cortisol shapes behavioural syndromes of juvenile coho salmon, Oncorhynchus kisutch. Social dominance during a territory intrusion and boldness following a simulated predator attack were recorded for juvenile coho salmon reared from cortisol-treated and untreated eggs. When exposed to a conspecific intruder, coho salmon treated with cortisol in ovo increased dominance behaviours, whereas coho salmon reared from untreated eggs reduced these behaviours. Following exposure to a simulated predator attack, coho salmon reared from cortisol-treated eggs increased boldness (i.e. increased activity and feeding; reduced shelter use), whereas coho salmon reared from untreated eggs reduced boldness. A dominance–boldness relationship was observed for coho salmon reared from cortisol-treated, but not untreated, eggs; dominance during the territory intrusion positively correlated with boldness under threat of predation. Our results highlight the complex influence of egg GCs on offspring behaviour and behavioural syndromes. Continued investigation into the outcomes of egg GCs on offspring will broaden our understanding of intergenerational components of the hormone–behaviour nexus.

Egg cortisol response to stress at early stages of development in Persian sturgeon Acipenser persicus

One of the most important periods in artificial breeding of sturgeon is incubation. In general, little is known about ontogeny of the stress response in early developmental stages of sturgeon. In this study, cortisol content was measured by radioimmunoassay for the first time in eggs of Persian sturgeon Acipenser persicus to elicit at which times during early developmental stages, that they were particularly sensitive to stressful events. In this study, fertilized eggs of mature Persian sturgeons were placed in special boxes (as replicates) that were put in incubation units of Yushchenko. To assess pre-stress samples (resting cortisol levels), 5 eggs were selected randomly from each box in 7 important stages of incubation (2-cell division, blastula, end of blastula, mid of gastrula, neurula, eyed stage and heart beaten). In another experiment, eggs were exposed to an acute stress (10 min out of water) in 7 developmental stages. For determination of post-stress cortisol, eggs were collected 2 and 6 h after the stress. The results indicated significant difference in various stages of incubation but cortisol levels revealed no differences in whole-body levels between stressed samples (2 and 6 h post-stress) and unstressed samples (P > 0.05). The highest percentage of hatching rate for post-stressed eggs was recorded in 2-cell division stage while this percentage declined steadily in the heart beaten stage when a statistically significant difference was revealed with regard to other stages (P = 0.000). These findings suggested that sturgeon eggs were not so much sensitive in their early developmental stages and relatively resistant to stress. However, significant decrease in survival rate in heart beaten stage showed a sensitivity of this stage to stress. Lack of an increase in cortisol after acute stress indicated that hypothalamus-pituitary-interrenal (HPI) axis for cortisol synthesis was not yet functional. However, further studies seem necessary to clarify when HPI axis is activated in this species.

Effects of maternal stress coping style on offspring characteristics in rainbow trout (Oncorhynchus mykiss)

Maternal size, age, and allostatic load influence offspring size, development, and survival. Some of these effects have been attributed to the release of glucocorticoids, and individual variation in these stress hormones is related to a number of traits. Correlated traits are often clustered and used to define the proactive and reactive stress coping styles. Although stress coping styles have been identified in a number of animal groups, little is known about the coupling between stress coping style and offspring characteristics. In the present study, plasma cortisol levels in ovulated mothers and cortisol levels in non-fertilized eggs from two rainbow trout (Oncorhynchus mykiss) strains selected for high (HR) and low (LR) post-stress plasma cortisol levels were compared. Offspring characteristics such as egg size, larval growth, and energy reserves also were compared between the two strains. Maternal plasma and egg cortisol levels were correlated, but no difference between the HR and LR strains was detected in either parameter. LR females produced larger eggs, and larvae with larger yolk sacs compared to HR females, however no differences in larval body size (excluding the yolk) was detected between strains. Considering that the HR and LR strains have a number of correlated behavioral and physiological traits that resemble the reactive and proactive stress coping styles, respectively, the results suggest that proactive mothers invest more energy into their offspring, producing larvae with larger energy reserves. It is possible that larger energy reserves in proactive larvae support the energy requirement for establishing and defending territory in salmonid fish. Furthermore, in the present study we found a positive relationship between mother plasma cortisol and egg cortisol; however neither mother plasma cortisol nor egg cortisol differed between strains. These results indicate that cortisol endowment from the mother to the offspring plays a minor role in the transfer of the behavioral and physiological traits which separates these strains.

The role of progeny quality and male size in the nesting success of smallmouth bass: integrating field and laboratory studies

Smallmouth bass display size-specific variation in reproductive success with larger brood-guarding males in a population more likely to rear offspring to independence than smaller individuals. The exact mechanisms responsible for this size-specific increase in reproductive output have yet to be identified. To assist in this process, we investigated the relationship between the size of brood-guarding male smallmouth bass and offspring quality (in this case, egg physiology, egg morphology, egg size, hatching success and lab survival). Further, we examined how factors such as egg physiology, egg morphology and egg size influenced reproductive success in the wild and hatching success in a controlled laboratory environment. Nesting male smallmouth bass that successfully reared their offspring to independence spawned earliest in the nesting period were the largest individuals, and guarded eggs with greater concentrations of cortisol compared to males that abandoned their offspring prematurely. Offspring survival in the laboratory was not correlated with offspring survival in the wild, indicating that caution should be used interpreting studies that attempt to relate laboratory-derived survival metrics to the wild. Together, results demonstrate size-specific differences in offspring quality for nesting smallmouth bass, which are correlated with higher concentrations of cortisol in eggs. However, hatching success under laboratory conditions was dissimilar to nesting success in the field relative to cortisol concentrations.

Egg hormones in a highly fecund vertebrate: do they influence offspring social structure in competitive conditions?

Summary1. Social status can vary considerably among individuals and has significant implications for performance. In addition to a genetic component, social status may be influenced by environmental factors including maternal effects such as prenatal hormone exposure. Maternal effects on traits determining social status have previously been examined in species where mothers provide parental care for relatively few offspring and therefore directly influence postnatal development. However, the generality of conclusions arising from these investigations is unclear because species that employ different reproductive strategies have not been studied.2. We investigated the hypothesis that egg steroid hormone levels influence the social status of juvenile brown trout (Salmo trutta). We manipulated intra‐clutch levels of cortisol and testosterone in eggs from 15 mothers using dilute hormone baths at the time of fertilization and examined their effects on traits that correlate with social status in juveniles [including standard body size, aggression, competitive ability and standard metabolic rate (SMR)].3. Hormone treatment did not affect whole‐animal or mass‐corrected SMR at the critical developmental stage when juveniles switch from reliance on a maternally provisioned yolk sac to independent feeding. However, juveniles from cortisol‐treated eggs were smaller at this developmental stage. They were also less aggressive than, and subordinate to, fish from untreated eggs in socially competitive conditions, even after correcting for the observed effect of cortisol on body size. Egg testosterone treatment resulted in a likely pharmacological or toxicological dose with subsequent effects on both body size and behaviour in independently feeding juveniles.4. Results from this study demonstrate that variation in the amount of cortisol deposited in eggs by spawning females influences juvenile social status and performance. The effects of elevated egg cortisol in fish are similar to the actions of embryonic glucocorticoids reported in other vertebrate taxa with very different reproductive strategies, suggesting a widespread mechanism for the effects of maternal stress on offspring. Possible adaptive aspects of this relationship are discussed.

In ovo elevated oocyte cortisol content induces innate immune responses in rainbow trout (Oncorhynchus mykiss) early embryonic cells

Event Abstract Back to Event In ovo elevated oocyte cortisol content induces innate immune responses in rainbow trout (Oncorhynchus mykiss) early embryonic cells Mao Li1*, Spencer Russell2, John S. Lumsden2 and John F. Leatherland1 1 University of Guelph, Biomedical Sciences, Canada 2 University of Guelph, Pathobiology, Canada In most fish embryos, the functional acquired immunity begins after hatch (1), and the protection of the embryos relies on the innate immunity of early embryo cells (1, 2). The immune suppressive effects of cortisol are well established in the developed organism, but the effects of maternal cortisol on early embryo immunity, and the ontogeny of innate immune components are not well established (2, 3). Lectins and TLRs were studied because they are widely distributed among fish tissues, and responsive to pathogens (5, 6). Naturally-ovulated rainbow trout oocytes were incubated in cortisol-enriched ovarian fluid for 3-h to examine the effects of the cortisol exposure on the ontogeny of innate immunity in rainbow trout embryos. Egg cortisol content was elevated from 4 ng/oocyte (controls - CC) to 5 (C1) and 7.5 ng/oocyte (C2) prior to fertilization (4). Lysozyme activity, intelectin, TLR-5M and TLR-5S gene expression were measured, and the intelectin and ladderlectin localization was examined in embryos using immunohistological [IH] methods. Lysozyme activity was significantly elevated in the C2 treatment group from the zygote until 13-days post fertilization (dpf), but was not affected at 21-dpf. Both intelectin and ladderlectin were present in 12-hours post-fertilization (hpf) (2-cell stage) embryos. But they had differences in response to cortisol exposure, intelectin levels (intensity of staining signal) were elevated in both cortisol treatment groups at 12-hpf and then suppressed between 36- and 48-hpf; ladderlectin was suppressed by both elevated cortisol treatments until 36-hpf compared to controls. Both lectins were distributed cell membranes and among epithelial tissues and brain tissues in late stage embryos. Intelectin mRNA was detected in oocytes and transcript levels were significantly elevated in both cortisol treatment groups relative to the controls after the 3-h incubation. There were no differences among the three treatment groups at 1- and 5-dpf, but intelectin mRNA levels were lower in the cortisol treatment groups at 13-dpf, and the transcript levels in the C2 treatment group were lower than in the CC and C1 treatment groups at 26-dpf. mRNA transcripts for the two TLR-5 genes were present in oocytes; transcript levels were significantly higher in both cortisol treatment groups relative to the controls after the 3 h incubation; TLR-5S mRNA was more abundant than that of TLR-5M mRNA. In conclusion, the ontogeny of the gene expression patterns, and the genes, lectins and lysozyme are response to increased oocyte cortisol content which indicates the importance of innate immunity in the early embryonic cells and embryo development prior to acquired immunity functions in fish. (Supported by NSERC and OMAF to JFL and JSL; NSERC Doctoral Scholarship to ML).

Effects of maternal stress on egg characteristics in a cooperatively breeding fish

Elevated stress experienced by a mother can compromise both her own reproductive success and that of her offspring. In this study, we investigated whether chronically stressed mothers experienced such effects in cooperatively breeding species, in which helpers at the nest potentially compound the negative effects of maternal stress. Using Neolamprologus pulcher, a group-living cichlid fish from Lake Tanganyika, we observed the effects of experimentally increased stress on female reproductive success (measured as inter-spawn interval, and number of eggs) as well as egg characteristics including egg size and cortisol concentrations. Stress levels were manipulated by repeated exposure to the acute stresses of chasing and netting. Stressed females had longer inter-spawn intervals and laid fewer, smaller eggs. Although no significant differences in egg cortisol concentrations were detected between control and stressed females, egg cortisol concentration fell between spawns in control but not in stressed fish. No effect of helper number was detected for any parameter examined, except there appeared to be less change in egg cortisol content in groups with helpers present. Our results suggest that stress imposes fitness costs on breeding females, and social regulation of a dominance hierarchy does not appear to exacerbate or alleviate the negative effects of maternal stress.

Plasma cortisol response to stress in juvenile rainbow trout is influenced by their life history during early development and by egg cortisol content

In this study, the consequences of the exposure of rainbow trout ( Oncorhynchus mykiss) to a brief stress during early development were explored on the later response of fingerlings to stress. Firstly, we analyzed the ontogeny of cortisol production and that of the initial cortisol response to stress in developing fish. It is only at the eyed stage that the embryos started to produce some basal cortisol. The HPI (hypothalamic–pituitary–interrenal axis) was however not functional before hatching, as exposure of the embryos to a stress did not trigger any cortisol response. A cortisol response to an acute stress was detected 9 days after hatching. In a second set of experiments, we showed that a very brief stress applied at 3 different early stages (eyed, hatching, and yolk resorption) reduced the later cortisol response to stress of 5-month-old fingerlings. This reduction is not likely to be due to alterations in the fish interrenal sensitivity because the 5-month-old fingerlings responded to ACTH treatment (only one dose tested) within the same magnitude as the fish that were not stressed during early development. An experimentally induced increase in egg cortisol just after fertilization also induced a reduction in stress sensitivity of 5 month old fingerlings, which was dose-dependent This study shows for the first time that the responsiveness of the corticotrope axis in 5–6 months old rainbow trout was influenced both by early stress exposure and by initial egg cortisol levels. Whether the HPI was functional or not at the time the initial stress was applied only had a small influence on the later unambiguous effect of early stress exposure.

Whole-body concentrations of cortisol and sex steroids in white sturgeon (Acipenser transmontanus, Richardson 1836) during early development and stress response

In general little is known about hormones and the ontogeny of the stress response in the early developmental stages of chondrostean fishes and in particular of white sturgeon (Acipenser transmontanus, Richardson 1836). In this study, we measured for the first time cortisol and sex steroids (testosterone and estradiol) in eggs, larvae, post-larvae, and fry of white sturgeon by radioimmunoassay (RIA), to elucidate some endocrine aspects of its development. The cortisol, testosterone, and 17β-estradiol of maternal origin found in unfertilized eggs of white sturgeon probably regulate both growth and development of the embryo. Cortisol decreased after fertilization, whereas testosterone and 17β-estradiol did not significantly change. During the late stages of embryo development and immediately after hatching, endogenous production of cortisol and sexual steroids, respectively, occurred. Sex steroids may be physiological inducers of gonad sex differentiation in sturgeon. All steroids showed an increase 10 days post-hatch (dph), near the transition from an endogenous to an exogenous energy source. Cortisol maintained the same basal levels even after metamorphosis, whereas testosterone and 17β-estradiol declined significantly in post-larvae at 35 and 45 days post-hatch. In addition, to evaluate the ontogeny of a functional hypothalamic-pituitary-interrenal (HPI) axis, larvae and fry were submitted to acute stress. The HPI axis did not seem to be functional on the first day post-hatch, but became so from the third day post-hatch onward.