The response to capture and confinement stress of plasma cortisol, plasma sex steroids and vitellogenic oocytes in the marine teleost, red gurnard

Reproductively active female red gurnard Chelidonichthys kumu were captured on long-lines, and placed in confinement tanks for 24, 48, 72 and 96 h to examine the effect of capture and confinement on reproductive parameters (experiment I). Plasma cortisol at the time of capture was elevated to levels typical of stressed fish in other species (53–125 ng ml⁻¹). Final plasma cortisol levels in red gurnard confined for any length of time were not significantly different from one another (ranging from 17 to 43 ng ml⁻¹), indicating that fish were chronically stressed when held in captivity for up to 96 h after capture. When initial and final plasma cortisol levels were compared within confinement groups, cortisol decreased significantly after 24 and 96 h of confinement indicating that some acclimation to captivity may have occurred. In contrast, plasma 17β-estradiol (E₂) and testosterone (T) levels decreased significantly to levels comparable to those in post-spawned fish, after any period of confinement, and remained low throughout the experiment. Another group of fish was captured and confined in the same manner as experiment I but subjected to repeated blood sampling every 24 h, until 96 h post-capture. In these fish, plasma cortisol levels decreased significantly from 127 ng ml⁻¹ after 24 h confinement and thereafter showed no change (25–45 ng ml⁻¹). Plasma E₂ decreased significantly after 72 h of confinement while plasma T showed no change from levels at capture. Increased amounts of follicular atresia were found in vitellogenic oocytes of fish confined for longer periods of time in experiment I, indicating that capture and confinement stress affect reproduction negatively in captive wild fish.     

Plasma cortisol concentrations before and after social stress in rainbow trout and brown trout

Two related experiments examined the relationship between plasma cortisol concentrations and the development of social hierarchies in fish. In the first, rainbow trout, Oncorhynchus mykiss, and brown trout, Salmo trutta, were observed for dominance interactions when confined within single-species pairs for 4, 48, or 168 h. Subordinate members of a pair exhibited significantly higher cortisol concentrations than dominant and single fish, but the pattern of cortisol elevation differed between the two species, being quicker to rise and increasing to a higher level in rainbow trout. Cortisol concentrations were correlated with behavioural measurements; the more subordinate the behaviour exhibited by a fish, the higher its cortisol concentration. Social stress was a chronic stressor, and no acclimation to social status occurred during the week. In the second experiment, measurements of plasma cortisol were made before pairing of rainbow trout and then after 48 h of confinement in pairs. Subordinate fish demonstrated significantly higher concentrations of plasma cortisol both before and after social stress. It therefore appears that in addition to cortisol being elevated during periods of social stress, an association may exist between initial cortisol levels and the likelihood of a fish becoming subordinate.     

Inhibition of spawning and associated suppression of sex steroid levels during confinement in the substrate-spawning Tilapia zillii

Substrate-spawning Tilapia zillii failed to spawn in crowded holding tanks but exhibited a marked tendency to spawn soon after transfer to individually partitioned aquaria. Confined conditions suppressed the levels of serum 17 β -oestradiol (E₂) and testosterone (T) in females, which remained low throughout the period of confinement. Levels of both steroids rose significantly following transfer of fish to individual aquaria and were maintained at consistently higher levels than those of fish which remained confined. Proportions of stage 3 (late perinucleolar) oocytes were significantly lower ( P <0.001) in individual fish 21–30 days after transfer, coincident with significantly higher ( P <0.01) proportions of stage 6 or 7 oocytes (late vitellogenic or maturing oocytes). However, no significant differences ( P ≥0.05) were detected between individual or confined groups of fish in the relative proportions of stage 2 (early perinucleolar), stage 4 (cortical alveolar) or stage 5 (early vitellogenic) oocytes. Atresia increased as the period of confinement increased. Following return of individual fish to confined conditions, blood steroids fell rapidly to levels seen in fish that had remained confined throughout. It is suggested that the reduced sex steroid levels detected during confinement were insufficient to allow developing oocytes to complete oocyte growth and maturation. The detection of significantly lower proportions of stage 3 and significantly higher proportions of stage 6/7 oocytes soon after transfer suggest that fish were preparing one batch of oocytes for spawning (stage 6/7 oocytes) whilst recruiting from a pool of previtellogenic oocytes (stage 3) for the ensuing spawning cycle.     

Relationship between stress, feeding and plasma ghrelin levels in rainbow trout, Oncorhynchus mykiss

Sexually immature rainbow trout were acclimated to small-volume (1 m³) holding tanks and then exposed to short-term stress to examine the relationship between feeding, stress, plasma ghrelin levels and other plasma stress parameters. Plasma ghrelin levels showed an increase 24 h after a single feed, plasma lactate and glucose levels decreased over the same period and plasma cortisol levels were low and constant. One hour of confinement stress resulted in elevations of plasma cortisol, glucose and lactate and depression of plasma ghrelin levels. In a separate experiment, 2 h of confinement stress also depressed feeding immediately after stress, concomitant with increases in plasma cortisol, lactate and glucose; however, in this case there was no change in plasma ghrelin concentrations. A repeat of the 2-h confinement experiment using fish that had not been acclimated to small-volume holding tanks produced a more marked elevation in plasma cortisol and a stronger suppression of feeding post-stress but in this case also, there was no change in plasma ghrelin levels. The results of this study confirm that feeding in rainbow trout is suppressed by confinement stress although the effect is transitory in this domesticated stock. Similar to that in other fishes, plasma ghrelin levels appear to be modulated by feeding status and may be influenced by stress, suggesting an orexigenic role for ghrelin in rainbow trout.     

Physiological and proteomic evidences that domestication process differentially modulates the immune status of juvenile Eurasian perch (Perca fluviatilis) under chronic confinement stress

The current study aimed to evaluate the influence of domestication process on the stress response and subsequent immune modulation in Eurasian perch juveniles (Perca fluviatilis) submitted to chronic confinement. Briefly, F1 and F4 generations were confined into small-size tanks and sampled 7 and 55 days after stocking. Cortisol and glucose levels as well as lysozyme activity and immunoglobulin level were evaluated in the serum. Spleen Somatic Index and spleen ROS production were also measured. A proteomic analysis was performed on serum sampled on day 7. Finally, both generations were genetically characterized using a microsatellite approach. Globally, results revealed that chronic confinement did not elicit a typical stress response but resulted in a prolonged immune stimulation. Proteomic results suggested that domestication process influenced the immune status of perch submitted to chronic confinement as the F1 confined fish displayed lower abundance of C3 complement component, transferrin and Apolipoprotein E. Microsatellite data showed a strong genetic drift as well as reduced genetic diversity, allelic number and heterozygosity along with domestication process. The present work is the first to report that fish under domestication can develop an immune response, assessed by a combined approach, following recurrent challenges imposed by captive environment despite a reduced genetic variation.     

Modulation of stress hormones in rainbow trout by means of anesthesia, sensory deprivation and receptor blockade

Sympathetic activation leading to increased levels of blood catecholamines, and stimulation of the hypothalamic-pituitary inter-renal axis leading to increased cortisol, are difficult to avoid when handling animals. Yet, in research on effects of acute stress, elicitation of such responses must be minimized in the control groups. The work examines means to achieve a minimally disturbed state in rainbow trout (Oncorhynchus mykiss). Level of arousal was determined by adrenaline and cortisol concentrations in plasma, and by the spleen:somatic index. Fish were prepared for bleeding by rapid capture and concussion, by infusion of anesthetic into the undisturbed home tank, by confinement in black boxes, or by being fed alpha- and beta-receptor antagonists. Even when done quickly, netting and concussion yielded fish with ca. 200-pmol adrenaline/ml plasma. Cortisol was elevated (to > 10 ng/ml) within 30 s of stress initiation. Surreptitious infusion of anesthetic (2-phenoxyethanol, PE) into tanks yielded fish with lower adrenaline levels (means 19.34 and 19.58 pmols/ml in home tank and black boxes, respectively). Among fish given phentolamine and propranolol, spleen:somatic indices and plasma adrenaline were higher than in diet controls, whether undisturbed or stressed, indicative of successful receptor blockade. Since careful infusion of 2-PE yielded the lowest adrenaline levels, and requires no special apparatus, it is the method of choice for obtaining minimally stressed fish.     

Stress does not inhibit induced vitellogenesis in juvenile rainbow trout

Vitellogenin (Vtg) is a widely used biomarker for xenoestrogen exposure in male fishes. In female fishes Vtg can be negatively affected by stress independent of declines in estrogen. However, few data are available on the effect of stress in male fish abnormally producing Vtg, such as when exposed to xenoestrogens. The objective for these studies was to determine the effects of stress on fish forced to produce Vtg. Three weeks prior to the experiment immature juvenile rainbow trout, Oncorhynchus mykiss, were acclimated to the experimental tanks and fed a maintenance ration. We induced Vtg synthesis by injecting 17β-estradiol (E₂) 7 days prior to experimentation. Treatments in duplicate tanks were: (1) no stressor; (2) stressor; (3) E₂; (4) E₂ and stressor. Plasma was collected at time = 0 for baseline measurements from eight fish per tank and Vtg was significantly elevated in treated fish compared to uninjected controls. Water was drained from the stressor tanks then refilled to a level that just covered the backs of the fish. Eight fish were sampled again at 4 and 9 h, and 1, 7, and 14 days of continuous stress. Stressor tanks were refilled with water to pre-stress levels and the fish were sampled after another 2 weeks. Cortisol was significantly elevated from the unstressed fish at 4 h; however, plasma Vtg in the E₂-stimulated fish was not affected by the stressor at any timepoint. These results indicate that fish capture procedures employed in the field or caging experiments likely do not lead to false negative results when plasma Vtg is used as a biomarker for xenoestrogen exposure. It also suggests that the energetic load induced by stress is insufficient to cause a reduction in Vtg, during a continuous E₂ administration, at least within the timepoints examined in this study.     

Effect of fasting on body composition and responses to stress in sunshine bass

The integrated responses of the hormonal regulation of growth and stress in sunshine bass (Morone chrysops X Morone saxatilis) as regulated by feed deprivation were investigated. Groups of fish were fed 1.5% of the body weight per day or offered no feed for 4 weeks. Another group of fish was not fed for 3 weeks and feed was offered during the fourth week. Fish in each group were sampled immediately before or after a 15-min low water confinement stressor after each week of the experiment. Liver mass and liver glycogen content were decreased after one week of fasting and remained low until the end of the study. However, both recovered after a week of refeeding. Intraperitoneal fat was significantly lower after two weeks of fasting and did not recover after a week of refeeding. None of these components were affected by confinement stress. Plasma glucose in unstressed fish was generally unaffected by fasting or refeeding; however, plasma glucose increased after confinement stress in fed but not in fasted fish. The cortisol stress response was unaltered by fasting and remained robust. Plasma IGF-I generally decreased in fasted fish but was not significantly lower than fed fish until the fourth week. A week of refeeding did not restore plasma IGF-I concentrations. Plasma IGF-I concentrations were higher in confinement stressed fed fish after two and four weeks but were unchanged in the fourth week. There was no change in the plasma IGF-I concentrations in fasted or refed fish due to the stress. Liver weight and liver glycogen were essentially depleted after 2 weeks of fasting. The reduction of liver glycogen greatly reduced the glucose response to stress; however, the cortisol stress response was maintained for at least four weeks of fasting. Intraperitoneal fat was decreased very little after 4 weeks of fasting. Plasma IGF-I concentrations were reduced only after 3 weeks of fasting.     

Changes in blood cortisol, glucose and lactate in carp retained in anglers'keepnets

Capture of carp Cyprinus carpio from holding tanks by dipnet, or from semi-natural conditions by rod and line, elicits a physiological stress response characterized by elevation of plasma cortisol levels. The transfer of carp to keepnets subsequent to capture does not increase or reduce the magnitude or duration of this response and in both cases plasma cortisol levels have returned to pre-stress levels within 24 h of the initial disturbance. The postcapture plasma cortisol elevation is accompanied by disturbances in plasma glucose and lactate levels but these are less consistent in severity and duration than the cortisol response. These data suggest that the retention of fish in keepnets following capture, does not represent a source of stress additional to that imposed by capture and has no effect on the rate of recovery of the fish from the initial capture stress.     

Ingestion and ejection of hooks: effects on long-term health and mortality of angler-caught yellowfin bream Acanthopagrus australis

Ninety juvenile yellowfin bream Acanthopagrus australis were angled from holding tanks, allowed to ingest nickel-plated, carbon-steel J-hooks and released (with their lines cut) into individual experimental tanks during 2 experiments in order to assess their (1) long-term (up to 105 d) health, mortality and rate of hook ejection and (2) short- and medium-term (< 42 d) temporal changes in health during hook ingestion. Equal numbers of control fish were scooped from holding tanks and similarly monitored in experimental tanks. Of 20 hook-ingested fish released during Expt 1, 3 died within 8 d, providing a non-significant mortality of 15%. Between Day 6 and Day 56 post-release, 13 of the surviving individuals ejected their hooks, which were typically oxidized to about 94% of their original weight and often broken into 2 pieces. At Day 105, there were no significant differences between the 20 control and 17 hook-ingested/-ejected fish in terms of their ability to digest and assimilate food (measured as changes in apparent digestibility coefficients), stress (measured as concentrations of plasma cortisol and glucose) or of morphological parameters that included weight (Wt) and maximum height (MH), maximum width (MW) and maximum girth (MG). During Expt 2, 3 individuals that still contained ingested hooks and 3 controls were sampled on each of 9 occasions between Day 3 and Day 42 post-release. All fish were sampled for blood cortisol and glucose and were then euthanized before being weighed and measured for total length (TL), MH, MW and MG. Hook-ingested individuals were also X-rayed to determine the position and orientation of hooks. There were no significant differences in plasma glucose between hook-ingested and control fish. Irrespective of the treatment of fish, concentrations of cortisol were elevated on some sampling occasions, indicating variable, acute stress. The MH and MG of fish were not significantly different between groups. Significant differences were detected for MG and Wt, with hook-ingested fish having weights similar to those of the control fish but a relatively greater MW (owing to stomach distension from ingested hooks) until 2 wk post-release, after which both morphological parameters generally declined. There was no significant temporal progression of hooks in the stomach of treatment fish; however, some hooks reorientated to positions that may have precluded passage along the digestive tract. We conclude that, for the J-hooks examined, cutting the line is an appropriate strategy that results in the greater majority of released hook-ingested yellowfin bream surviving with minimal negative long-term effects.     

Analysis of stress-induced hepatic gene expression in rainbow trout (Oncorhynchus mykiss) selected for high- and low-responsiveness to stress

The production and welfare of intensively reared fish would be improved by reducing stress responsiveness. One approach to achieving this goal is selective breeding utilising stress-responsive genes as direct genetic markers of the desirable trait. As a first step in this process, microarray analysis has been carried out on liver tissues of rainbow trout selectively bred for high (HR) or low (LR) responsiveness to a stressor. Microarray hybridizations provided gene expression profiles for pooled samples of fish confined for 6 h, 24 h and 168 h and for individual fish (168 h only). 161 genes were shown to be differentially regulated in HR and LR fish during confinement exposure and eight of these gene expression profiles were validated by quantitative PCR. Genes of particular interest included intelectin-2 precursor which showed greater than 100-fold higher expression in HR fish compared to LR fish irrespective of whether the fish were confined or not; interferon inducible transmembrane protein 3 which was differentially stress-induced between the two lines; and hepatic pro-opiomelanocortin B (POMC B) which was upregulated during stress in HR fish but downregulated in LR fish. All these offer potential as direct markers of low stress responsiveness in a marker-assisted selection scheme.     

Stress-associated impacts of short-term holding on fishes

Most sources of stress in aquaculture, fish salvage, stocking programs, and commercial and sport fisheries may be unavoidable. Collecting, handling, sorting, holding, and transporting are routine practices that can have significant effects on fish physiology and survival. Nevertheless, an understanding of the stressors affecting fish holding can lead to practices that reduce stress and its detrimental effects. The stress-related effects of short-term holding are influenced by water quality, confinement density, holding container design, and agonistic and predation-associated behaviors. Physiological demands (e.g., resulting from confinement-related stresses) exceeding a threshold level where the fish can no longer compensate may lead to debilitating effects. These effects can be manifested as suppressed immune systems; decreased growth, swimming performance, or reproductive capacity; even death. Furthermore, holding tolerance may depend upon the species, life stage, previous exposure to stress, and behavior of the held fish. Water quality is one of the most important contributors to fish health and stress level. Fish may be able to tolerate adverse water quality conditions; however, when combined with other stressors, fish may be quickly overcome by the resulting physiological challenges. Temperature, dissolved oxygen, ammonia, nitrite, nitrate, salinity, pH, carbon dioxide, alkalinity, and hardness are the most common water quality parameters affecting physiological stress. Secondly, high fish densities in holding containers are the most common problem throughout aquaculture facilities, live-fish transfers, and fish salvage operations. Furthermore, the holding container design may also compromise the survival and immune function by affecting water quality, density and confinement, and aggressive interactions. Lastly, fishes held for relatively short durations are also influenced by negative interactions, associated with intraspecific and interspecific competition, cannibalism, predation, and determining nascent hierarchies. These interactions can be lethal (i.e., predation) or may act as a vector for pathogens to enter (i.e., bites and wounds). Predation may be a significant source of mortality for fisheries practices that do not sort by size or species while holding. Stress associated with short-term holding of fishes can have negative effects on overall health and well-being. These four aspects are major factors contributing to the physiology, behavior, and survival of fishes held for a relatively short time period.     

Dynamics of viral hemorrhagic septicemia, viral erythrocytic necrosis and ichthyophoniasis in confined juvenile Pacific herring Clupea pallasii

Capture of wild, juvenile herring Clupea pallasii from Puget Sound (Washington, USA) and confinement in laboratory tanks resulted in outbreaks of viral hemorrhagic septicemia (VHS), viral erythrocytic necrosis (VEN) and ichthyophoniasis; however, the timing and progression of the 3 diseases differed. The VHS epidemic occurred first, characterized by an initially low infection prevalence that increased quickly with confinement time, peaking at 93 to 98% after confinement for 6 d, then decreasing to negligible levels after 20 d. The VHS outbreak was followed by a VEN epidemic that, within 12 d of confinement, progressed from undetectable levels to 100% infection prevalence with >90% of erythrocytes demonstrating inclusions. The VEN epidemic persisted for 54 d, after which the study was terminated, and was characterized by severe blood dyscrasias including reduction of mean hematocrit from 42 to 6% and replacement of mature erythrocytes with circulating erythroblasts and ghost cells. All fish with ichthyophoniasis at capture died within the first 3 wk of confinement, probably as a result of the multiple stressors associated with capture, transport, confinement, and progression of concomitant viral diseases. The results illustrate the differences in disease ecology and possible synergistic effects of pathogens affecting marine fish and highlight the difficulty in ascribing a single causation to outbreaks of disease among populations of wild fishes.     

A cohabitation challenge to compare the efficacies of vaccines for bacterial kidney disease (BKD) in chinook salmon Oncorhynchus tshawytscha

The relative efficacies of 1 commercial and 5 experimental vaccines for bacterial kidney disease (BKD) were compared through a cohabitation waterborne challenge. Groups of juvenile chinook salmon Oncorhynchus tshawytscha were vaccinated with one of the following: (1) killed Renibacterium salmoninarum ATCC 33209 (Rs 33209) cells; (2) killed Rs 33209 cells which had been heated to 37 degrees C for 48 h, a process that destroys the p57 protein; (3) killed R. salmoninarum MT239 (Rs MT239) cells; (4) heated Rs MT239 cells; (5) a recombinant version of the p57 protein (r-p57) emulsified in Freund's incomplete adjuvant (FIA); (6) the commercial BKD vaccine Renogen; (7) phosphate-buffered saline (PBS) emulsified with an equal volume of FIA; or (8) PBS alone. Following injection, each fish was marked with a subcutaneous fluorescent latex tag denoting its treatment group and the vaccinated fish were combined into sham and disease challenge tanks. Two weeks after these fish were vaccinated, separate groups of fish were injected with either PBS or live R. salmoninarum GL64 and were placed inside coated-wire mesh cylinders (liveboxes) in the sham and disease challenge tanks, respectively. Mortalities in both tanks were recorded for 285 d. Any mortalities among the livebox fish were replaced with an appropriate cohort (infected with R. salmoninarum or healthy) fish. None of the bacterins evaluated in this study induced protective immunity against the R. salmoninarum shed from the infected livebox fish. The percentage survival within the test groups in the R. salmoninarum challenge tank ranged from 59% (heated Rs MT239 bacterin) to 81% (PBS emulsified with FIA). There were no differences in the percentage survival among the PBS-, PBS/FIA-, r-p57- and Renogen-injected groups. There also were no differences in survival among the bacterin groups, regardless of whether the bacterial cells had been heated or left untreated prior to injection.     

Ventilatory frequency of Nile tilapia subjected to different stressors

Ventilatory frequency (VF) was investigated in the fish Nile tilapia, Oreochromis niloticus, subjected to confinement, electroshock or social stressor. Fish were allowed to acclimatize to tank conditions for 10 days (1 fish/aquarium). VF baseline was determined 5 days after adjustment had been started. At the 10th day of isolation, stressor effects on VF were assessed. The stressors were imposed during 60 min: pairing with a larger resident animal (social stressor), or gentle electroshock (AC, 20 V, 15 mA, 100 Hz for 1 min every 4 min), or space restriction (confinement). VF was quantified immediately after the end of the stressor imposition. Baseline of VF was not statistically different among groups. Social stressor clearly induced VF to increase, while an increased or decreased VF can be observed for both confinement and electroshock. However, fish tend to increase their VF in response to confinement and decrease in the case of electroshock. These results suggest that VF is a sensitive behavioural indicator for distinguishing stress responses in the fish Nile tilapia among different stressors.     

Pathogenesis of acute ulceration response (AUR) in hybrid striped bass

In a previous study, we discovered that acute confinement stress causes rapid ulceration of the fins of hybrid striped bass Morone saxatilis female x M. chrysops male (Noga et al. 1998. Vet Pathol 35:102-107). In this paper, we report the development of a reproducible model for studying this phenomenon in juvenile hybrid striped bass. We also determined how quickly ulceration could develop in acutely stressed fish and documented the sequential light microscopic and ultrastructural changes associated with this response. When hybrid striped bass were subjected to a standardized confinement protocol, the pathological response was extremely rapid (fin ulceration began to develop within 15 min of confinement). Grossly, the distal edges of the fins became blanched, and melanophores aggregated near the basement membrane and dermis after 15 min of confinement. Microscopically, the earliest detectable change in the fins, which occurred within 15 min of confinement, was swelling and loss of microridges of the outermost epidermal cells; this was followed by epidermal erosion. After 30 min of stress, epidermal ulceration developed at the distal edges of the fins. At this time, both necrotic and apoptotic epidermal cells were present. The middle and basal epidermal layers were severely spongiotic and the dermis and hypodermis were edematous. Over longer periods (up to 2 h), lesions were similar but increasingly more severe, progressing from the distal edge of the fin towards the base. The response to acute stress showed a significant correlation between confinement period and severity of the pathological changes (epidermal degeneration, epidermal ulceration and leukocyte infiltration). Also, we demonstrated that epidermal damage was not restricted to the fins but also affected the body skin and eyes. The ventral area of the body and the corneal epithelium of stressed fish were ulcerated; however, skin on the head and operculum was not affected, suggesting a site-specific mode of damage. In stressed fish, epidermal ulceration was found in 67 to 97% of all fins, 88% of skin on the ventrum, and 67% of corneas, while control fish had only very mild epidermal ulceration in the few fish in which it was present (on 5 to 10% of the fins, but not on the ventral skin or corneas). Due to the widespread damage to epidermal tissues of the body surface, we have named this the acute ulceration response (AUR). Our study indicates that acute confinement can rapidly cause significant damage to epidermal and ocular epithelium. AUR might be a primary cause of morbidity in acutely stressed fish.     

Juvenile growth and aggression in diploid and triploid Chinook salmon Oncorhynchus tshawytscha (Walbaum)

Multilocus heterozygosity, aggressive and feeding behaviour, plasma cortisol levels and growth rate were evaluated among three groups of juvenile Chinook salmon Oncorhynchus tshawytscha : diploid, triploid and mixed groups of diploid and triploid fish. There was no difference between diploid and triploid fish in measurements of heterozygosity calculated using seven microsatellite loci, and these measurements did not correlate with performance measurements including feeding rate and growth rate. Aggression trials that examined small groups of fish revealed that after 4 days together in tanks, triploid fish were significantly less aggressive during feeding than diploid fish or fish in mixed groups. At the end of the trials, however, plasma cortisol levels did not differ among the three groups. Thirty-day growth trials in duplicate tanks of 60 fish revealed no difference in growth rate among diploid, triploid and mixed groups, but plasma cortisol levels were significantly lower in triploid fish than in either diploid fish or the mixed fish. Overall, independent of the above differences in aggressive behaviour and cortisol levels, these results suggest similar performance in diploid and triploid Chinook salmon, and thus provide support for the viability of triploid Chinook salmon culture in commercial aquaculture.     

Cohesive social behaviour shortens the stress response: the effects of conspecifics on the stress response in lake sturgeon Acipenser fulvescens

An examination was made of whether social interactions can have a beneficial effect through the attenuation of the stress response in a social species. In the first experiment, one larger (mean ± s . e . 194·0 ± 12·5 g) and seven smaller (32·0 ± 2·6 g) juvenile lake sturgeon Acipenser fulvescens were placed in tanks to determine whether a classic dominance effect would be established based on body size ( n = 6). Large fish did not establish a territory or aggressively interact with smaller fish, as there were no significant differences in nearest-neighbour distances and an absence of aggressive behaviour (biting, chasing and pushing). In the second experiment, it was hypothesized that the presence of conspecifics would have a beneficial effect through an attenuation of the stress response. Fish in groups or isolation were stressed by a brief aerial exposure (30 s), and blood plasma was measured at regular time intervals (0, 20, 40, 60, 120 and 240 min) following the stressor via an implanted cannula ( n = 9–11). The presence of conspecifics did not affect the peak cortisol response, however, the overall cortisol response was shorter in duration compared to fish in isolation. Furthermore, secondary stress variables (plasma ions and glucose) showed differences between fish in groups and isolation. The results of these experiments suggest that social interaction plays an important and beneficial role in regulating the stress response in cohesive social species such as A. fulvescens .     

Effects of confinement stress and additional zinc treatment on some blood parameters in the dogfish Scyliorhinus canicula

Several blood parameters were studied in the dogfish Scyliorhinus canicula subjected to different confinement conditions (transport and maintenance) and zinc contamination. Confinement stress significantly decreased erythrocyte counts (RBCC), haematocrit (Ht), haemoglobin (Hb), leucocrit (Lt), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC). Mean corpuscular volume (MCV) did not change with this stress and plasma glucose concentration significantly increased. In fish subjected to confinement stress conditions, Zn treatment significantly decreased Hb, MCH and MCHC and plasma glucose concentration. Ht and MCV did not change and Lt and RBCC significantly increased. Results are discussed related to other stress effects and effects of zinc treatment on fish confined in non-stressful conditions.     

Dietary intake of β-1,3/1,6 glucans in juvenile dentex (Dentex dentex), Sparidae: effects on growth performance, mortalities and non-specific defense mechanisms

Juvenile common dentex ( Dentex dentex ) Sparidae, were fed moist diets over 2 months. The three test groups (triplicate lay out) received diets supplemented every second week with one of the two tested immunostimulants — MakroGard or Vita-Stim, or a diet without supplement (control). All fish were weighed at weekly intervals to (a) obtain data on growth and feed conversion and (b) generate repeatedly a defined stress situation to amplify the possible effects of the immunostimulant treatments on growth, mortality and non-specific defense mechanisms.
Growth and feed conversion were not significantly influenced by the applied immunostimulants during the entire experimental period. Most likely, as a result of the cumulative handling stress, growth decreased considerably during weeks 5 and 6 of the experiment in all three test groups. Fish in five of the nine test tanks developed a protozoan infection, regardless of the immunostimulant applied. However, mortalities related to this disease were markedly reduced in tanks treated with either MacroGard (10%) or VitaStim (15%) compared to the control tank (30%). Mortalities related either to cannibalism, eye damage or other unidentified events were not significantly different between treated and control tanks. At the end of the experiment, general fish health parameter (Haematocrit, Leucocyte number) and parameter of the non-specific immune response (Oxygen radical activity, Lysozyme activity, Total plasma protein) were determined from blood samples. Blood parameter did not clearly reflect the protective effect of the administered immunostimulants.