Validation of enzyme-linked immunosorbent assay for measurement of faecal cortisol in fish

Quantification of glucocorticoid (GC) levels in faeces has become an established method for the non-invasive assessment of adrenocortical activity. These hormones are frequently determined in plasma samples as parameters of adrenal activity and response to stress. Because GCs are metabolized and excreted with both intact hormone and their metabolites present in faeces, the concentration of GCs can be measured in excreta. Faecal samples present the advantages of easy collection, no stress to the animal and elimination of the issue of potentially misleading acute GC spikes. The aim of this study was to determine if an enzyme-linked immunosorbent assay (ELISA) for cortisol was appropriate for monitoring adrenocortical activity in faecal casts of fishes. Performance of the cortisol ELISA was validated by comparison to high-performance liquid chromatography, which is an established method for measuring free GCs and GC metabolites in faeces. Parallelism and sample extraction efficiency were compared for the two methods. Pearson's correlation across samples for these methods was 0.996. Results demonstrated that the ELISA was an efficient, sensitive and reliable method for cortisol measurement in faecal extracts, which should permit integration of non-invasive stress monitoring into studies of fish behaviour and physiology.     

Noninvasive assessment of nitrate‐induced stress in koi Cyprinus carpio L. by faecal cortisol measurement

This study determined effects of nitrate (NO₃^?)‐induced stress on faecal cortisol levels in koi (Cyprinus carpio). NO₃^? was presented in two formats: (1) bulk increase followed by dilution to assess cortisol response and recovery and (2) bulk increase followed by incremental increases to assess cortisol response limit. Fish were maintained group‐wise in treatment and control aquaria (n=6 per group), and 0.5 g L^?1 NO₃^? (as NaNO₃) was added to the water. Faecal samples were collected daily and blood samples were taken pretreatment and 72‐h posttreatment and were assayed for cortisol via ELISA. NO₃^? increased plasma and faecal cortisol 4.4‐fold and 3.9‐fold respectively. Plasma cortisol was not measured further. After a 74% NO₃^? decrease by dilution, faecal cortisol decreased to baseline within 24 h, and restimulation by NO₃^? (1.0 g L^?1) elevated faecal cortisol to maximal study levels. In a separate experiment, exposure to 0.4 g L^?1 NO₃^? increased faecal cortisol 6.9‐fold. However, three additional 0.4 g L^?1 NO₃^? increments across 9 days did not further increase cortisol. This study demonstrates that faecal cortisol measurement in fish via an ELISA can be useful as an indicator of NO₃^?‐induced stress in the aquatic environment.     

β-adrenergic signal transduction in fish: interactive effects of catecholamines and cortisol

The elevation of plasma catecholamine levels during acute stress initiates a series of compensatory physiological and biochemical mechanisms to alleviate the disruptive effects of stress on blood oxygen transport. Of particular importance is the β-adrenergic activation of a Na⁺/H⁺ antiporter associated with the red blood cell (rbc) membrane. Upon activation, the Na⁺/H⁺ antiporter extrudes H⁺ from the rbc and the resultant alkalinization of the rbc interior serves to enhance both the affinity and the capacity of haemoglobin O₂ binding. The activation of the Na⁺/H⁺ exchanger is dependent upon the intracellular accumulation of cyclic AMP. The extent of cyclic AMP accumulation is determined, in part, by the number and/or affinities of cell surface β-adrenoreceptors. Recent studies have shown that the number of cell surface β-adrenoreceptors are rapidly increased during acute hypoxia and that this phenomenon may explain the enhanced responsiveness of hypoxic rbc's to exogenous catecholamines. In certain instances, plasma catecholamine and cortisol levels rise concurrently. We recently have shown that chronic (10 day) elevation of cortisol levels, in vivo, or short-term (24h) elevation, in vitro, caused significant elevation of internalized β-adrenoreceptors. Upon exposure of the rbc's to hypoxia, these additional receptors are rapidly recruited to the cell surface where they become functionally coupled to adenylate cyclase. Ultimately, therefore, chronic elevation of plasma cortisol levels increases the responsiveness of the rbc to circulating catecholamines. We recently have identified similar enhancement of cell surface β-adrenoreceptors by cortisol and increased physiological responsiveness (glycogenolysis) to catecholamines in trout hepatocytes. Thus, chronic elevation of cortisol levels appears to be generally adaptive for increasing the sensitivity of the β-adrenergic signal transduction system of at least two cell types (rbc's, hepatocytes) involved in the amelioration of acute stress when plasma catecholamine levels rise.

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Résumé L'élévation des niveaux de cathécolamines plasmatiques pendant un stress aigu déclenche une série de mécanismes physiologiques et biochimiques de compensation destinés à suprimer les effets destructeurs du stress sur les transport d'oxygène par le sang. L'activation β-adrénergique de l'antipore Na⁺/H⁺ associé à la membrane des globules rouges du sang (rbc) est particulièrement importante. Après activation, l'antipore Na⁺/H⁺ excrète du globule rouge H⁺ et l'alcalinisation du milieu intérieur du globule qui en résulte sert à stimuler l'affinité et la capacité de liaison hémoglobine-O₂. L'activation de l'échangeur Na⁺/H⁺ est dépendante de l'accumulation intracellulaire d'AMP cyclique. La quantité d'AMP cyclique accumulée est déterminée en partie par le nombre et/ou l'affinité des β-adrénorécepteurs situés à la surface des cellules. Des études récentes montrent que le nombre de β-adrénorécepteurs de surface augmente rapidement pendant une hypoxie aigue et que ce phénomène pourrait expliquer la stimulation de la réponse des globules rouges hypoxiques à des cathécolamines exogènes. Dans certains cas, les niveaux plasmatiques de cortisol et de cathécolamine augmentent simultanément. Nous avons récemment montré qu'une augmentation chronique (10 jours) des niveaux de cortisol in vivo ou une élévation de courte durée (24h) in vitro conduisent à une augmentation des β-adrénorécepteurs internalisés. Suite à l'exposition de globules rouges à une hypoxie, ces nouveaux récepteurs sont rapidement recrutés à la surface de la cellule où ils deviennent fonctionnellement couplés à l'adénylate cyclase. En dernier lieu donc, une élévation chronique des niveaux de cortisol augmente la réponse des globules rouges aux cathécolamines circulants. Nous avons récemment identifié une augmentation similaire des β-adrénorécepteurs de surface cellulaire par le cortisol et une réponse physiologique (glycogénolyse) stimulée par les cathécolamines dans les hépatocytes de truite. Ainsi, une élévation chronique des niveaux de cortisol semble être en général adaptative et augmentant la sensibilité du système de transduction du signal β-adrénergique dans au moins 2 types de cellules (les globules rouges et les hépatocytes) impliquées dans l'amélioration du stress aigu quand les niveaux de cathécolamine plasmatique augmentent.

     

Alternative protein blends as a replacement for fish meal in diets for white seabass,Atractoscion nobilis

An 8‐week feeding trial was conducted to determine the effectiveness of replacing fish meal (FM) with blends of alternative proteins in diets for white seabass (WSB, Atractoscion nobilis) at a starting weight of 5.6 g. Five diets were formulated with 400–440g kg^?1 crude protein (380g kg^?1 digestible). These included a high 520g kg^?1 FM control diet, a series of three diets with a sequential replacement of FM containing 410g kg^?1, 510g kg^?1 and 630g kg^?1 of a soy‐based protein blend (SPC) and 200g kg^?1, 100g kg^?1 and 0g kg^?1 FM, respectively and a fifth diet containing 550g kg^?1 of a corn‐based protein blend (CGM) and 100g kg^?1 FM. Survival was highest in the FM control group at 99% but all other performance measures (weight gain, feed conversion ratio, specific growth rate and protein retention efficiency) were worse than the other treatment groups. Weight gain reached a maximum of 595% in the SPC 200g kg^?1 FM treatment group. Performance decreased as inclusion of the soy‐based protein blend increased. The CGM treatment performed comparably to the SPC 100g kg^?1 FM treatment among all measures, except for survival, which was higher in the CGM 100g kg^?1 FM treatment. With nutrient levels and alternative protein blends used in this study, FM can be reduced to 100g kg^?1 of the diet for WSB without reductions in performance.     

拥挤胁迫后鲫鱼血液皮质醇和溶菌酶水平的变化及对病原的敏感性

分别在拥挤胁迫后第3、10、20、30天对鲫鱼(Carassiusauratus)血液皮质醇水平和血液溶菌酶水平进行了检测,并在每次采集血样后用嗜水气单胞菌(Aeromonashydrophila)进行人工感染以评估鱼体抵抗力。结果显示,血液皮质醇水平在胁迫后几天内显著升高(P<0.005),随后虽有所下降,但至第30天高密度组仍保持较高水平(216.32±11.91)ng/mL。实验初期,血液溶菌酶水平小幅升高,但经过长时间的拥挤后,高密度组血液溶菌酶活性受到极大抑制,显著低于同期对照组(P<0.01)。攻毒后的死亡率与胁迫时间成正相关,在最后一次采样期,高密度组死亡率高达58.3%,而同期对照组仅为16.7%,两者具统计学差异(P<0.01)。实验表明,经过拥挤胁迫后,鲫鱼血液皮质醇水平和溶菌酶水平均发生了变化,短期内,两者成正相关,而长期胁迫则表现为负相关。另外,拥挤胁迫也引起了鱼体抵抗力的下降。     

Recent developments and improvements in ornamental fish packaging systems for air transport

Current ornamental fish packaging systems are characterized by very high fish loading densities and high metabolic wastes in the transport water after shipment. They focus mainly on management of the quality of transport water. Recent studies using the guppy as a model fish showed that post‐shipment mortality could be reduced through enhancement of the stress resistance of the fish, and hence emphases should also be placed on the preparation of the fish for transport and recovery of the fish after shipment. Farmers can contribute significantly by applying nutritional prophylaxis before harvesting. Exporters may use the salinity stress test to identify fish lots of good quality for transport, apply health prophylaxis to eradicate parasites and optimize other techniques such as starvation of the fish or addition of salt to the transport water to enhance the stress resistance of the fish. Importers may adopt proper acclimation procedure and allow fish to recover in low salinity water to reduce post‐shipment mortality. As the main bulk of post‐shipment mortality is stress‐mediated and occurs during the 1‐week recovery period, the industry should consider revising the basis of the current warranty system for their customers, from death on arrival to cumulative mortality at 7 days post shipment (or death after 7 days, DA7), in order to cut down fish losses after shipment.     

Stress responses and disease resistance in salmonid fish: Effects of chronic elevation of plasma cortisol

Basal levels of plasma cortisol in unstressed salmonid fish are normally in the range 0–5 ng ml⁻¹. An acute stress such as handling or 1 h confinement caused a temporary elevation of the plasma cortisol levels of both brown trout,Salmo trutta L., and rainbow trout,Salmo gairdneri Richardson, in the range 40–200 ng ml⁻¹ with a return to basal levels within 24–48 h. The extent of the cortisol elevation in response to an acute stress was dependent upon both the species and strain of trout. Chronic stresses, such as prolonged confinement or crowding, resulted in an elevation of plasma cortisol levels to approximately 10 ng ml⁻¹. Under these circumstances, blood cortisol levels remained elevated for periods of up to 4 weeks before acclimation finally occurred. It is shown, by means of intraperitoneal implantation of cortisol, that chronic elevation of plasma cortisol levels in the brown trout results in a dose-dependent increase in mortality due to common bacterial and fungal diseases. This effect is apparent at plasma cortisol levels as low as 10 ng ml⁻¹, levels below those often reported as being representative of ‘unstressed’ fish. These findings are discussed in relation to the known immunosuppressive effects of corticosteroids in teleost fish.     

Effect of acid environments on cortisol and cortisol receptor activity in Atlantic salmon,Salmo salar

The cytosol and nuclear extract of gill tissue obtained from laboratory held Atlantic salmon,Salmo salar manifested saturable cortisol binding of high affinity and low capacity (cytosol: K_a = 0.198 ± 0.024 × 10⁹/M, N_max = 116.8 ± 20.8 fmol/mg protein; nuclear extract: K_a = 0.823 ± 0.057 × 10⁷/M, N_max = 1563 ± 330 fmol/mg protein; n = 4). The cytosol receptor activity displayed high steroid and tissue specificity and a single binding peak at 191,000 Da following gel permeation chromatography. Atlantic salmon exposed for 3 or 8 months to waters from the Medway River (pH about 5.1), the Westfield River (pH about 4.8) and calcium carbonate treated Westfield River (pH about 5.6) showed no gill cytosol receptor activity. Cortisol receptor activity in the gill nuclear extracts from fish in limed Westfield River water in December (3 months) was less than half the activity in the fish treated with Medway River water (p < 0.05) although the plasma cortisol values were not different. In May (8 months), the plasma cortisol of fish in limed water was almost twice that of the fish held in acid Westfield River water (p = 0.058).     

A behavioural sensor for fish stress

Due to water turbidity, fish stress might be difficult to observe. Evaluation of fish stress by blood sampling requires removing a fish from the water, which is in itself a stressful event. Therefore, we designed and built a sensor to detect fish behaviour that reflects stress. The electronic sensor detected early signs of fish stress by scoring the fish's inactivity. LEDs and detectors are embedded on a steel wand that is held underwater by an operator. In this preliminary (feasibility) study, the new sensor was validated for Tilapia (Cichlidae) and Hybrid Striped Bass (Morone). We induced stressful situations in the fish tanks by manipulating oxygen and temperature levels.ResultsLowering the temperature and oxygen levels both significantly increased the average number of signals identified by the sensor, which indicate stress. The effect of reducing water temperature from 24 °C to 15 °C was three times stronger than was the effect of lowering the oxygen saturation level from 85% to 50%. The difference in the number of signals between the good and stressful conditions was statistically significant, amounting to approximately eight sensor signals, 10.57 compared to 2.49 respectively. Lowering the temperature increased the mean number of signals by 5.85 and 6.06 at 85% and 50% oxygen saturation respectively, whereas lowering oxygen levels increased the mean number of signals by 2.02 and 2.23 at 24 °C and 15 °C, respectively. The results indicate that the stress status of cultured fish can be evaluated using the proposed behavioural sensor. The new sensor may provide an earlier indication of a problem in a fish tank or pond than was heretofore possible. This early warning can enable the fish farmer to take action before many fish are harmed.     

Experimental investigation on the transport of different fish species in a jet fish pump

A jet fish pump with a throat of ø60 mm was designed to study its performance in the transport of different fish species and the physiological changes in fish thereafter. Experiments were conducted to investigate the fish transport rate and energy required to transport each ton of fish when transporting Carassius auratus, commonly known as the Chinese goldfish, Megalobrama amblycephala, or Wuchang bream, and Ctenopharyngodon idella, the grass carp. Fish were examined for external injuries as well as for several important enzymes and hormones which are indicators of tissue injury and stress. The results showed that the transport rate for all three species of fish rose dramatically with an increase in the primary stream rate. In this experiment, the transport rates of C. auratus, M. amblycephala and C. idella reached 2357 ± 37.2 kg ∙ h⁻¹, 2888 ± 41.6 kg ∙ h⁻¹, and 2060 ± 40.2 kg ∙ h⁻¹, respectively. However, both injury rate and energy required to transport each ton of fish increased no matter whether the primary stream rate was too low or too high. Considering both transport rate and injury rate, the mean primary stream rate of 80 m³ ∙ h⁻¹ was determined to be the optimal operating condition in this experiment. Fish were stressed and most likely some of their organs were damaged. However, most physiological indexes almost fully recovered after several hours.     

High stocking density produces crowding stress altering some physiological and biochemical parameters in gilthead seabream, Sparus aurata, juveniles

High stocking density has been shown to produce a wide variety of effects on cultured fish populations, such as alterations in behavior and poor feed utilization, resulting in mortality and poor growth. High stocking density has also been reported to produce chronic stress and mobilization of energy sources in fish. There are few studies focusing on the effect of high stocking density on one of the most important marine fish species for Mediterranean aquaculture, gilthead seabream (Sparus aurata). Consequently, this study investigates the effect of high stocking density on juveniles of this species, focusing on effects of stocking density on growth, biochemical composition, immune status and hematology.The results of this study showed that high stocking density produced a chronic stress situation. Plasma cortisol levels increased up to 16.25 ng ml–1 in fish held at high stocking; this value was 4 fold that of fish held at low stocking; density (3.91 ng ml–1 ). As a consequenceof this stressful situation, there was both haemoconcentration and a decrease in alternative complement pathway (ACP), an important component of the immune system of fish (from 167.23 U ml–1 down to 146.37 U ml–1). Haematocrit, haemoglobin concentration and red blood cell count were significantly higher in fish held at high stocking density (43.87%, 10.76 9dl–1 and 3.36 ×106 mm–3, respectively) compared with those fish held at low stocking density (37.21%, 9.32 g/dlg–1 and 2.82 ×106 mm–3, respectively).In addition, high stocking density produced a decrease in hepatosomatic index (from 2.26 down to 2.04) and altered liver fatty acid composition. Oleic acid (18: 1n-9) decreased in liver total lipids of fish held at high stocking density and arachidonic acid (20: 4-n6) and n-3 high unsaturated fatty acids (n-3 HUFA) were reduced in liver polar lipids of those fish. These alterations reflect the effect of stocking density on lipid metabolism to help meet the increased energy demand.     

Method for underwater measurement of the auditory brainstem response of fish

ABSTRACT:   The auditory brainstem response (ABR) of fishes is commonly measured by bringing the heads of the fishes out of the water in a small tank. However, this method is inapplicable to experiments for large fishes that are economically important in large spaces such as the sea or in a large tank. This paper describes a method of recording, the ABR for fishes in water, without exposing the fish heads to air, by using a waterproof, insulated electrode. To evaluate the effectiveness of this method, the goldfish Carassius auratus was investigated, and the ABR waveform and auditory thresholds measured in water were compared with those measured on the surface. Both ABR waveforms and auditory thresholds showed similar trends between the two methods. The underwater ABR method is useful to measure the auditory thresholds of larger fish in natural or on-site environments such as the sea, net enclosures and large aquaria in which precise positioning of the fish is not possible. However, more improvement is needed to apply this method to large fishes.     

Ontogeny of the cortisol stress response in yellow perch (Perca flavescens)

The ontogeny of the cortisol stress response in yellow perch (Perca flavescens) was documented by measuring resting and stress-induced whole-body cortisol levels in fish at several early development stages. In newly fertilized eggs resting cortisol levels averaged 41.0 ± 3.0 pg/embryo, and decreased to 8.0 ± 3.0 pg/embryo by day 6 where they remained constant until hatch on day 10. Acute stress challenge tests (30 sec cold shock) conducted on embryonic (days 6 and 8 post-fertilization) and larval (hatch and 2 days post-hatch) yellow perch revealed no differences in whole-body cortisol levels between stressed and unstressed individuals. In acute stress challenge tests (30 sec net confinement) conducted on post-hatch yellow perch, differences in cortisol levels between stressed (30 min post-stress) and unstressed fish were detected one week after hatching (12.3 ± 1.6 ng/g and 7.0 ± 1.3 ng/g, respectively). The magnitude of the stress response was greater in 2- to 5-week-old fish than in one-week-old fish. Changes in post-stress cortisol levels were closely associated with changes in interrenal morphology. Cumulatively, these results indicate that cortisol biosynthesis, integration and maturation of the hypothalamic-pituitary-interrenal (HPI) axis can be observed in yellow perch as early as one week after hatching. This revised version was published online in July 2006 with corrections to the Cover Date.     

Docosahexaenoic acid (C22:6n‐3) alters cortisol response after air exposure in Prochilodus lineatus (Valenciennes) larvae fed on enriched Artemia

This study investigated the effect of different proportions of dietary docosahexaenoic acid (DHA) and arachidonic acid (ARA) on the growth, survival and stress response of curimba (Prochilodus lineatus Valenciennes) larvae, an omnivorous freshwater species, fed on enriched Artemia. Six different experimental emulsions were formulated to have two levels of ARA (A‐0.5 and A‐1.0), two levels of DHA (D‐0.5 and D‐1.0) two combinations of ARA and DHA (AD‐3:1 and AD‐1:3). There were no significant effects of treatments on curimba standard length and survival at the end of the experiment. The results showed a negative correlation between dietary DHA and whole‐body cortisol after 60 s of air exposure. The resting concentration of cortisol was not affected by the diets (p = .424). After air exposure, all treatments except for D‐1.0 (p = .137) and D‐0.5 (p = .545) exhibited a marked cortisol rise. These results showed that DHA supplementation can have an attenuating effect on cortisol response after air exposure.     

Endocrinology and fish culture

In the current practice of fish culture the use of hormones is mainly limited to the field of reproduction, and more precisely to induce or synchronize ovulation and stimulate spermiation. The practice of pituitary homogenates injection (called hypophysation) which started in the early 1930's has allowed spectacular developments in the culture of some cyprinid species, especially in China, India and Europe. HCG has been used successfully in a limited number of species and LHRH-A, often associated with antidopaminergic compounds, started to be used in some species, especially cyprinids, on a commercial scale. Sex steroids are now commonly used to reverse the sex of some species in salmonids, cyprinids, tilapias. Due to legal restriction and consumer opposition, the tendency is to limit the use of steroids, especially in fish which are later offered to consumption. GH, which significantly stimulates growth and reduces the food conversion rate in laboratory experiments, will possibly be used on a large scale in fish farms. Endocrinology has considerably increased the knowledge of the mechanisms controlling some physiological functions; this has allowed the identification of sensitive phases in fish in captivity, and helps in the management and the process of domestication (most of the cultured fish species have been taken form the wild recently).     

Individual variations in behavior and free cortisol responses to acute stress in tiger pufferfish Takifugu rubripes

Abstract:   The aim of the present study was for individual variation in behavior to be characterized and related to differences in physiological stress responses in juvenile tiger pufferfish (fugu) Takifugu rubripes . A set of siblings ( n  = 330) from wild parents was subjected to behavior tests, and some were classified into active and inactive groups ( n  = 42 each group). The active animals consisted of those individuals that kept active swimming during 6 min after transfer from one tank to another, and the inactive animals were those that stopped swimming within 6 min. Time-course acute stress responses, which were elicited by anesthesia and subsequent removal of a pectoral fin, were compared between the active and inactive fish. The stress indicators used were free and total plasma cortisol, and plasma glucose. Although no significant difference was observed at each sampling point between groups, the inactive fish required shorter recovery time from acute stress in all three indicators than the active fish. The percentage of free to total plasma cortisol in the pre-stress condition was significantly lower in the inactive group than in the active group. Our findings indicate that fugu has behavioral variation that is associated with stress responsiveness. The inactive fish are expected to be more tolerant of stress and suitable for aquaculture.     

The use of clove oil, metomidate, tricaine methanesulphonate and 2-phenoxyethanol for inducing anaesthesia and their effect on the cortisol stress response in black sea bass (Centropristis striata L.)

Juvenile and adult black sea bass (Centropristis striata L.) were exposed to various concentrations of four anaesthetics to determine practical dosages for handling as well as for procedures such as bleeding, ovarian biopsy or tag implantation. In experiment 1, juveniles exposed to either 2.0 mg L^?1 metomidate, 15 mg L^?1 clove oil, 70 mg L^?1 tricaine methanesulphonate (TMS) or 200 mg L^?1 2‐phenoxyethanol (2‐PE) reached stage II of anaesthesia in 3–5 min and could be handled for weighing and measuring. All fish had completed recovery to stage III within 6 min. In experiment 2, the established concentrations of each anaesthetic were tested on juveniles to determine their ability to prevent a reflex to a subcutaneous needle puncture. All of the fish exposed to clove oil (20 mg L^?1) and 40% of the TMS‐treated (70 mg L^?1) fish reacted while none of the fish anaesthetized in metomidate (2.0 mg L^?1) or 2‐PE (200 mg L^?1) responded to the needle puncture. In experiment 3, metomidate (5.0 mg L^?1), clove oil (30 mg L^?1) TMS (125 mg L^?1) or 2‐PE (300 mg L^?1) were all effective for performing an ovarian biopsy or tag implantation on adults. In experiment 4, TMS (125 mg L^?1) exacerbated the cortisol response to a short handling stressor during a 30 min exposure. Fish anaesthetized in 2‐PE (300 mg L^?1), metomidate (5.0 mg L^?1) or clove oil (40 mg L^?1) had increased cortisol levels associated with the handling stressor but there were no further increases during the remainder of the experimental period. The results demonstrate that these anaesthetics are effective for sedation and anaesthesia of black sea bass and that the best choice is dependant upon the procedures to be performed.     

Whole-body corticosteroid and plasma cortisol concentrations in larval and juvenile Atlantic cod Gadus morhua L. following acute stress

Methods were developed to assess whole‐body immunoreactive corticosteroid concentrations (IRC) and plasma levels of cortisol in Atlantic cod subjected to several common, acute stressors. A measurable corticosteroid stress response was observed at the first sampling in whole bodies of 8‐day post‐hatch (dph) larvae. Two groups of juveniles (～5 and 30 g) were subjected to a 30 s net stressor and whole‐body IRC and plasma cortisol levels were determined. Post‐stressor IRC in smaller fish rose approximately 14‐fold, peaked at 1 h, were sustained for 3–6 h and returned to pre‐stressor levels within 24 h. Post‐stressor plasma cortisol levels in larger fish rose approximately 18‐fold, peaked at 0.5–1 h, were sustained for 1–3 h and then returned to near pre‐stressor levels after 24 h. Immunoreactive corticosteroid concentrations appeared to remain elevated longer than plasma cortisol levels suggesting that steroids other than cortisol were contributing to total immunoreactivity in homogenates. Juveniles exposed to either a grading procedure or high density transport had maximal IRC and plasma cortisol levels within 90 min which returned to pre‐stressor levels within 24–72 h. A reduction in water salinity (20 g L^?1) did not moderate the corticosteroid response in juveniles. The results show that Atlantic cod respond to common, acute stressors in a manner similar to other teleosts. Whole‐body homogenates can be used to identify changes in IRC in response to acute stress in cod with the caveat that recovery IRC may differ from plasma cortisol concentrations.     

A microcomputer program for stimulating effects of physical transport processes on fish larvae

A simple microcomputer program uses Lagrangian particle tracking to simulate the fates of individual larvae subject to wind- and tide-driven advection fields and diffusion. Space/time advection patterns are provided to the program as inputs; thus, the program can use results from various hydrodynamic models. The program can simultaneously track several groups of larvae that are assigned varying attributes, including (1) body size, to allow simulation of effects of variation in growth rates; (2) spawning locations and times; (3) vertical migration behavior; and (4) settlement strategy (duration of larval period, depth conditions for settlement). The model is intended as an exploratory tool to help identify alternative hypotheses that might explain observed life history patterns and causes of inter-annual variation in recruitment rate. For English sole in the Hecate Strait, B.C., the model leads us to hypothesize that observed spawning locations have resulted from a tradeoff between places that would provide the best feeding opportunities and places that would minimize risk of advective export to unfavorable rearing habitats.     

Stress hormones and the cellular stress response in salmonids

The relationship between stress protein (SP) levels and the hormonal stress response in salmonids was examined through the measurement of gill SP70 and SP30 levels together with plasma cortisol, glucose and ion concentrations, in response to handling stress (45 s holding in a net), intraperitoneal cortisol implants, and heat shock (+10 °C). Handling and cortisol implants resulted in increased plasma cortisol and glucose levels. Heat shock following handling reduced plasma [Na⁺] below that observed in response to the handling stress alone, and heat shock following cortisol implant significantly lowered both plasma [Cl^–] and [Na⁺] below that of fish receiving the cortisol implant alone. Increased SP70 levels occurred 1 h following the 2 h +10 °C heat shock. Handling the fish prior to the application of heat shock suppressed the increase of SP70 levels in the gills. However, increased plasma cortisol concentrations alone did not attenuate gill tissue SP70 increase caused by heat shock. Physiological (10^–7 M) and pharmacological (10^–5 M) concentrations of adrenaline caused increased levels of SP70 in hepatocytes. Addition of the -blocker propanolol blocked this response to adrenaline. The results indicate that handling procedures do not result in an increase of hsp30 or hsp70 and may suppress hsp synthesis under certain circumstances.