Stress in African catfish (Clarias gariepinus) following overland transportation

Of the many stressors in aquaculture, transportation of fish has remained poorly studied. The objective of this study was therefore to assess the effects of a (simulated) commercial transportation on stress physiology of market-size African catfish (Clarias gariepinus). Catfish weighing approximately 1.25 kg were returned to the farm after 3 h of truck-transportation, and stress-related parameters were measured for up to 72 h following return. Recovery from transportation was assessed through blood samples measuring plasma cortisol, glucose and non-esterified fatty acids (NEFA) and gill histology. Also, the number of skin lesions was compared before and after transport. Pre-transport handling and sorting elevated plasma cortisol levels compared to unhandled animals (before fasting). Plasma cortisol levels were further increased due to transportation. In control fish, plasma cortisol levels returned to baseline values within 6 h, whereas it took 48 h to reach baseline values in transported catfish. Plasma glucose and NEFA levels remained stable and were similar across all groups. Transported catfish did not, on average, have more skin lesions than the handling group, but the number of skin lesions had increased compared to unhandled animals. The macroscopic condition of the gills was similar in control, transported and unhandled catfish; however, light microscopy and immunohistochemistry revealed atypical morphology and chloride cell migration normally associated with adverse water conditions. From our data, we conclude that transportation may be considered a strong stressor to catfish that may add to other stressors and thus inflict upon the welfare of the fish.     

Effects of chronic confinement on physiological responses of juvenile gilthead sea bream, Sparus aurata L., to acute handling

Understanding how gilthead sea bream, Sparus aurata L., an important Mediterranean Sea species for aquaculture, respond physiologically to stressors commonly encountered in intensive rearing is important for effective production, as managing for stress is a major factor in maintaining healthy fish stocks. Our objective was to determine whether holding juvenile gilthead sea bream at a high density (HD), as a chronic stressor, would affect their physiological responses to a subsequent acute handling stressor. After acclimation at a low density (LD) of 6 kg m^?3 in 200‐L circular tanks containing 33–36 g L^?1 recirculating seawater at 19°C under a normal photoperiod, juvenile 37‐g gilthead sea bream were confined for 14 days at a HD of 26 kg m^?3 and then subjected to 30‐s aerial emersion in a dipnet. Plasma levels of cortisol, glucose, lactate, osmolality and chloride were determined in fish held in separate lots during LD (control) and HD confinement at 0, 1, 2, 7 and 14 days, and then after handling at 0, 1, 2, 4 and 8 h. Although plasma cortisol levels were similar in LD and HD fish groups after 14 d of confinement (15 and 23 ng mL^?1, respectively), the cortisol response in fish from the HD treatment at 1 and 2 h following acute handling (70 and 37 ng mL^?1, respectively) was only about half of that measured in the control group (139 and 102 ng mL^?1); plasma cortisol was similar in both groups by 4 and 8 h. In contrast, plasma glucose elevations in response to handling were higher at 4 and 8 h in the HD‐held fish (94 and 72 mg dL^?1, respectively) than in those from the LD treatment (59 and 51 mg dL^?1); glucose responses were similar in both groups at 1 and 2 h after handling and throughout confinement. Plasma lactate levels were higher in LD fish than in the HD group at the beginning of the experiment but were similar after 14 d confinement and responses to handling were similar (e.g. 33 and 35 mg dL^?1 at 1 h). Plasma osmolality showed increases during the first 2 h after acute handling but no differences were evident between the two density treatments at any time during confinement or posthandling. Plasma chloride levels did not change throughout the experiment. The reduced plasma cortisol response to acute handling likely resulted from negative feedback of mildly but chronically elevated circulating cortisol caused by the confinement stressor on the hypothalamic–pituitary–interrenal axis. While other post‐handling physiological changes also showed differences between treatment groups, the suppressed cortisol response in the HD‐held fish suggests a reduction in the gilthead sea bream's normal capacity to respond to an acute stressor.     

Cortisol and finfish welfare

Previous reviews of stress, and the stress hormone cortisol, in fish have focussed on physiology, due to interest in impacts on aquaculture production. Here, we discuss cortisol in relation to fish welfare. Cortisol is a readily measured component of the primary (neuroendocrine) stress response and is relevant to fish welfare as it affects physiological and brain functions and modifies behaviour. However, we argue that cortisol has little value if welfare is viewed purely from a functional (or behavioural) perspective—the cortisol response itself is a natural, adaptive response and is not predictive of coping as downstream impacts on function and behaviour are dose-, time- and context-dependent and not predictable. Nevertheless, we argue that welfare should be considered in terms of mental health and feelings, and that stress in relation to welfare should be viewed as psychological, rather than physiological. We contend that cortisol can be used (with caution) as a tractable indicator of how fish perceive (and feel about) their environment, psychological stress and feelings in fish. Cortisol responses are directly triggered by the brain and fish studies do indicate cortisol responses to psychological stressors, i.e., those with no direct physicochemical action. We discuss the practicalities of using cortisol to ask the fish themselves how they feel about husbandry practices and the culture environment. Single time point measurements of cortisol are of little value in assessing the stress level of fish as studies need to account for diurnal and seasonal variations, and environmental and genetic factors. Areas in need of greater clarity for the use of cortisol as an indicator of fish feelings are the separation of (physiological) stress from (psychological) distress, the separation of chronic stress from acclimation, and the interactions between feelings, cortisol, mood and behaviour.     

Effects of density,PVC‐tubes and feeding time on growth,stress and aggression in African catfish (Clarias gariepinus)

Successive exposure to aquaculture‐related stressors may compromise the allostatic capacity of African catfish and lead to allostatic overload and poor welfare. Therefore, we tested the effect of (i) feeding during the light or dark phase, (ii) density (51 fish per 140 L versus 51 fish per 43 L) and (iii) altered available resting space/shelter on stress handling were studied in African catfish (Clarias gariepinus). Available resting space/shelter was manipulated by providing PVC‐tubes in the tanks. Growth, baseline stress load (cortisol and osmolality), metabolism (glucose, lactate and non‐esterified fatty acids) and aggression (measured by skin scar incidence) were assessed. Upon completion of the experimental period, we determined the adaptive capacity of the fish by the response to air exposure. We show that night‐feeding enhances growth and lowers feed conversion ratio compared to day‐feeding. No effects of density were found for baseline stress hormones, basal metabolism or aggression. Low density seems to stimulate aggressive behaviour after air exposure. Unexpectedly, providing PVC‐tubes under high‐density conditions increased aggression, raised baseline cortisol levels and decreased basal metabolism as well as growth; also the endocrine response to air exposure was stronger. We argue that these effects relate to a reduction in available space, i.e. a further increase in density, or to an increased competition for shelter. Overall, our data suggest that night‐feeding optimizes growth and that care should be taken in using objects as shelter/enrichment for African catfish.     

Effects of Pro-Tex on zebrafish (Danio rerio) larvae,adult common carp (Cyprinus carpio) and adult yellowtail kingfish (Seriola lalandi)

Aquaculture practices bring several stressful events to fish. Stressors not only activate the hypothalamus–pituitary–interrenal-axis, but also evoke cellular stress responses. Up-regulation of heat shock proteins (HSPs) is among the best studied mechanisms of the cellular stress response. An extract of the prickly pear cactus (Opuntia ficus indica), Pro-Tex, a soluble variant of TEX-OE^®, may induce expression of HSPs and reduce negative effects of cellular stress. Pro-Tex therefore is used to ameliorate conditions during stressful aquaculture-related practices. We tested Pro-Tex in zebrafish (Danio rerio), common carp (Cyprinus carpio L.) and yellowtail kingfish (Seriola lalandi) exposed to aquaculture-relevant stressors (thermal stress, net confinement, transport) and assessed its effects on stress physiology. Heat shock produced a mild increase in hsp70 mRNA expression in 5-day-old zebrafish larvae. Pro-Tex increased basal hsp70 mRNA expression, but decreased heat-shock-induced expression of hsp70 mRNA. In carp, Pro-Tex increased plasma cortisol and glucose levels, while it did not affect the mild stress response (increased plasma cortisol and glucose) to net confinement. In gills, and proximal and distal intestine, stress increased hsp70 mRNA expression; in the distal intestine, an additive enhancement of hsp70 mRNA expression by Pro-Tex was seen under stress. In yellowtail kingfish, Pro-Tex reduced the negative physiological effects of transport more efficiently than when fish were sedated with AQUI-S^®. Overall, our data indicate that Pro-Tex has protective effects under high levels of stress only. As Pro-Tex has potential for use in aquaculture, its functioning and impact on health and welfare of fish should be further studied.     

Growth performance and physiological changes of holding juvenile Siberian sturgeon at different sizes in rearing units

An investigation was accomplished to find the effects of different sizes as a social stress on physiological responses and growth performance of Siberian sturgeon. Juvenile Siberian sturgeon were separated into two sizes (860.88 ± 22.32 g as larger fish and 427.28 ± 2.97 g as smaller fish). Three treatments including large size (L), small size (S), and mixed size (M) were considered. There were equal numbers of large and small fish in the M treatment. Sampling was carried out at 1st week (as a short‐term sampling) and 8th week (as a long‐term sampling) of the experiment. At the end of each sampling, haematocrit, glucose, cholesterol, triglyceride, total protein, lactate, and cortisol were measured. All growth parameters and feed utilization in large fish were not affected in the M treatment compared with the L treatment and also small fish in the M treatment compared with the S treatment. The final biomass in the S treatment was significantly higher than those in the L and M treatments. Haematocrit, glucose, cholesterol, triglyceride, total protein, lactate, and cortisol in the 1st week showed no significant difference with different sizes. However, in the 8th week, only haematocrit increased (by time not size) in the S treatment. Fish in the M treatment in both sizes did not experience the stress condition. Larger and smaller sizes had a positive social interaction; therefore, different sizes of Siberian sturgeon can maintain together without decreasing growth rate and also adverse physiological responses.     

Effect of blood glucose level on acute stress response of grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

Stress has a considerable impact on welfare and productivity of fish, and blood glucose level of fish may be a factor modulating stress response. This study evaluated the effect of blood glucose level and handling on acute stress response of grass carp Ctenopharyngodon idella. Fish were intraperitoneally injected with glucose at 0, 0.2, 0.5, and 1.0 mg g^?1 body mass (BM) and then were exposed to handling for 5 min. Glucose injection resulted in increase of plasma glucose level and liver glycogen content and decrease of plasma lactate level. Handling resulted in increase of plasma levels of cortisol, glucose, and lactate and plasma lactic dehydrogenase (LDH) activity and decrease of liver glycogen content. At 1 h post-stress, the plasma cortisol level was lower in the stressed fish injected with glucose at 0.5 mg g^?1 BM than the stressed fish injected with glucose at 0, 0.2, and 1.0 mg g^?1 BM. No significant differences were found in the activities of phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate kinase (PK) in the liver between the stressed and unstressed fish, regardless of the dose of glucose injection. At 1 h post-stress, the liver glucose-6-phosphatase (G6Pase) activity was higher in the fish without glucose injection than in the fish injected with glucose. This study reveals that blood glucose level can affect stress response of grass carp by modulating cortisol release and glucose homeostasis through glycogen metabolism and gluconeogenesis in the liver.     

Effects of iodized feed on stress modulation in steelhead trout, Oncorhynchus mykiss (Walbaum)

In the aquaculture industry, the physiological systems of fish can be chronically stressed by various biological, chemical and physical factors. Chronic stress leads to a decrease in the overall health and growth of the fish, making them more prone to diseases. Dietary iodine has been shown to reduce this stress response in chickens and increase disease resistance in dairy cattle, but the mechanisms by which iodine affects stress resistance and immunocompetency is not completely understood and has not been extensively studied in fish. This study investigated the effects of iodized feed as a nutritional supplement in relation to stress modulation in steelhead trout (Oncorhynchus mykiss). We investigated effects on primary, secondary and tertiary stress responses by measuring plasma levels of cortisol, glucose and thyroid hormones as well as haematocrit percentage and average growth rate in steelhead trout. Iodine‐supplemented fish, on average, had lower levels of plasma cortisol and glucose and lower packed cell volumes than fish fed with regular commercial feed (P<0.05). Iodine‐supplemented fish also showed higher levels of thyroid hormones and exhibited better growth over the period of the experiment (P<0.05). It was concluded that, used with other husbandry practices, use of iodine‐supplemented feed could lead to better host defence, growth and survival in fish raised in the aquaculture industry.     

The response of coral trout (Plectropomus leopardus) to capture, handling and transport and shallow water stress

The stress response of coral trout (Plectropomus leopardus ) to wild capture or controlled shallow water stressors was investigated. Stress associated with capture from the wild, handling and transport and shallow water evoked significant changes in circulating levels of cortisol, glucose, lactate, haemoglobin (Hb) and haematocrit (Hct) in coral trout. Plasma glucose, Hb and Hct increased over 30 to 60 min in response to the controlled stressor and then returned toward unstressed levels, even if the stressor persisted. Cortisol increased to maximum levels over the 60 min following the onset of the stressor and remained elevated for 4 h in the case of a single 30 min shallow water event or 3 d in response to the capture, handling and transport. The difference between the responses of blood glucose and cortisol suggests that cortisol does not maintain blood glucose during stress in coral trout. The concentrations of circulating thrombocytes, lymphocytes and granulocytes were also significantly affected by shallow water stress. As lymphocytes and granulocytes are important immune components, reduced concentrations of these cells may explain the increases in disease susceptibility, commonly observed in stressed fish during live transport.     

Suppression of Stress Responses in Siberian Sturgeon,Acipenser baeri,Juveniles by the Dietary Administration of Bovine Lactoferrin

A study was performed to investigate the influence of dietary lactoferrin (LF) on growth and stress responses of juvenile Siberian sturgeon. Fish were fed six experimental diets (0, 100, 200, 400, 800, and 1600 mg LF/kg diet) for 10 wk. At the end of the experiment, fish were exposed to an acute stressor of 2‐min air exposure. The levels of cortisol, glucose, and lactate were measured before and at 1, 3, 6, 9, 12, and 24 h after stress. LF supplementation did not influence the fish growth. Cortisol and lactate, but not glucose level, were significantly affected by dietary LF. Cortisol levels in the control group rose twofold 1 h after stress and were significantly higher than that in LF groups. Moreover, the lowest level of cortisol at all sampling times (prestress and poststress) was observed in fish fed 400 mg LF/kg diet. Lactate concentrations in sturgeon fed 400 mg LF/kg markedly rose 1 h after stress, albeit changes in other LF groups were not significant. Results of this study show that dietary LF can suppress the stress responses in Siberian sturgeon, while 400 mg LF/kg diet appears to be an effective dietary level for this species to reduce stress.     

Chronic stress due to high stocking density in open sea cage farming induces variation in biochemical and immunological functions in Asian seabass (Lates calcarifer,Bloch)

Stocking density is an important factor in cage aquaculture of finfish. Effects of high stocking density (35 fish cubic m^?1) on a range of biochemical and immunological parameters in Asian seabass reared in open sea floating net cages were compared to fish held in relatively low density (15 fish cubic m^?1). The results revealed that chronic stress due to high stocking density induced variations in most of the parameters studied as evidenced by increased cortisol and glucose levels and decreased activity of lysozyme, myeloperoxidase and complement. Production of reactive oxygen species, total leucocyte count and total serum protein were also decreased, whereas anti-protease, alkaline phosphatase and acid phosphatase activities were increased in high stocking-density group when compared to low stocking-density group. Effects of chronic stress due to high stocking density were discussed in relation to variations in these parameters.     

Effect of stocking density on growth and serum concentrations of thyroid hormones and cortisol in Amur sturgeon, Acipenser schrenckii

This study investigated the effects of different stocking densities on growth and serum concentrations of thyroid hormones and cortisol in Amur sturgeon, Acipenser schrenckii. Fish were reared at low, medium, and high stocking densities (initial experimental densities were 0.30, 0.75, and 1.78 kg m⁻², respectively) for 70 days. The results showed that high stocking density had negative effects on growth and feeding efficiency, and altered serum levels of thyroid hormones and cortisol in Amur sturgeon. A significant decrease in specific growth rate was observed as stocking density was increased. The feeding rate decreased significantly in the medium and high density groups, indicating that high stocking density reduced the food consumption of sturgeon. Food conversion ratio increased with increasing stocking density, suggesting that high stocking density might inhibit fish growth through decreasing food conversion efficiency. Serum concentrations of total triiodothyronine, free thyroxine, and free triiodothyronine were inversely related to stocking densities, whereas serum total thyroxine level of sturgeon stocked at different densities remained stable. Also, higher stocking density resulted in an elevation of serum cortisol level, indicating that the sturgeon stocked at the higher density experienced density-dependent physiological stress. These results suggest growth suppression caused by high stocking density might be related to both crowding stress and the declines in peripheral circulating levels of thyroid hormones, as well as associated with the reductions in both food consumption and food conversion efficiency.     

Sex-related differences in the organismal and cellular stress response in juvenile salmon exposed to treated bleached kraft mill effluent

Exposure of fish to stressors can elicit biochemical and organismal changes at multiple levels of biological organization collectively known as stress responses. The organismal (plasma glucose and cortisol levels) and cellular (hepatic hsp70) stress responses in fish have been studied in several species, but little is known about sex-related differences in these responses. In this study, we exposed sexually immature juvenile chinook salmon (Oncorhynchus tshawytscha) to bleached kraft mill effluent (BKME: 0%, 1%, and 10% v/v) for 30 days and then measured components of their organismal and cellular stress responses. Males exposed to 1% BKME had higher levels of plasma glucose than females. Plasma cortisol levels were unaffected in females exposed to BKME, but males exposed to 10% BKME had significantly higher levels of plasma cortisol relative to non-exposed males. While exposure to BKME did not affect hsp70 levels in males, females exposed to 1% BKME had higher levels of hsp70 relative to non-exposed and 10% BKME groups. Within any given treatment, females had higher levels of hsp70 relative to males. This study demonstrates that sex-related differences exist in commonly used indicators of stress in fish, and points out the importance of considering the sex of the fish in stress research. This revised version was published online in July 2006 with corrections to the Cover Date.     

Physiological stress responses in spotted wolffish (Anarhichas minor) subjected to acute disturbance and progressive hypoxia

The spotted wolffish (Anarhichas minor) is considered a promising species for coldwater aquaculture. It is a sedentary, bottom-dwelling fish which exhibits a calm and “non-stressed” behaviour in captivity. There are, however, no reports on the physiological responses to stressors in this species. In the present study we investigated primary (cortisol secretion) and secondary (glucose mobilization) responses to common aquaculture stressors like disturbance and hypoxia. Pre-stress plasma cortisol levels were within those generally considered representative for unstressed fish (~ 10 ng ml^− 1), whereas basal glucose levels were unusually low (0.3–0.4 mM). After exposure to a short, but severe disturbance challenge (emptying the tank of water for 10 min), the increase in plasma cortisol level was slow and relatively weak, reaching a peak level of 25 ng ml^− 1 4 to 8 h after disturbance. When the fish were exposed to a gradually decreasing oxygen level in the tank, a significantly elevated plasma cortisol level (35 ng ml^− 1) was seen in the fish that remained in the tank until oxygen saturation had decreased to 20% oxygen saturation after 2.5 h. However, a two-fold higher plasma cortisol level (~ 70 ng ml^− 1) was seen in all fish exposed to reduced oxygen levels (60, 40 and 20% oxygen saturation) after 3.5 h recovery in normoxic water. Plasma glucose levels showed only moderate increases (~ 70%) following disturbance and hypoxia challenges. An in vivo injection of ACTH caused a strong elevation of plasma cortisol (peak level ~ 170 ng ml^− 1), demonstrating a high capacity for interrenal steroidogenesis in the spotted wolffish. The slow and relatively weak cortisol response to stressors, and low plasma glucose levels, may relate to the sedentary lifestyle of the spotted wolffish. The stress-response is characterised by a passive (reactive) coping style, which is considered adaptive for farming of this species.     

Physiological changes in male and female pikeperch Sander lucioperca (Linnaeus, 1758) subjected to different photoperiods and handling stress during the reproductive season

Pikeperch broodstocks were exposed to different photoperiods: constant light (24L:0D), constant darkness (0L:24D), and 12 h light, 12 h darkness (12L:12D), for 40 days. Half of the broodstocks of each photoperiod were exposed to handling stress at a specific time of the day. Results showed that cortisol and lactate did not reveal any significant difference. However, glucose levels in females increased in the stress-free darkness period in comparison with stressful darkness photoperiods (0L:24D-s). Red blood cells in males and white blood cells in females showed a significant difference under different photoperiod regimes. Both sexes showed no significant difference in the differential count of leukocytes under different photoperiods and handling stress. Constant photoperiods and handling stress affected the hematological parameters, particularly, the number of lymphocytes and neutrophils in females. Our findings revealed that due to a long-term exposure to stressors, pikeperch brooders become adapted to stressful conditions.     

Diel changes in plasma cortisol and effects of size and stress duration on the cortisol response in European sea bass (Dicentrarchus labrax)

European sea bass (Dicentrarchus labrax), one of the most economically important fish in Mediterranean mariculture, shows high basal cortisol concentrations compared with other teleosts. The present study aims (a) to identify cortisol diel variation in fish held under a 12L:12D cycle and minimum handling stress, and (b) to establish the effect of fish size and stressor duration on the cortisol response. The results indicate high intrapopulation variability in plasma cortisol and a significant diel fluctuation with a peak value at dusk (18 h). Stressors of different intensity and/or duration affected the cortisol stress response in a differential manner according to fish size (and/or age). Maximum cortisol values in small-size fish were found at 1 and 2 h post-stress, depending on the duration of the stressor, while at 0.5 h post-stress in large fish regardless stress duration.     

Influence of immunostimulant polysaccharides,nucleic acids,and <Emphasis Type="Italic">Bacillus</Emphasis> strains on the innate immune and acute stress response in turbots (<Emphasis Type="Italic">Scophthalmus maximus</Emphasis>) fed soy bean- and wheat-based diets

Immunostimulants are widely applied in aquaculture practice and may have beneficial effects on the immune system and physical functions allowing higher tolerance to stress. In the current study, the impact of four (i–iv) dietary active ingredients on the immune and stress response of turbot was examined in two experiments (I and II). A basal low fish meal (FM; 32%) diet was formulated and supplemented with (i) yeast β-glucan and mannan oligosaccharide (GM), (ii) alginic acid (AC), (iii) yeast nucleotides and RNA (NR), or (iv) Bacillus strains (BS). The basal diet (C-LF) and a high FM (59%) control (C-HF) were maintained. All six diets were fed to juvenile turbots for 84 days in experiment I and for additional 28 days prior to experiment II. Immunological and hematological parameters were determined in experiment I. In experiment II, physical stress response to a typical short-term (<1 day) aquaculture handling procedure (combination of capture, netting/transfer, and crowding) was investigated. For this, turbot blood was sampled before and at 0.5, 1, 4, and 24 h post stress. Plasma lysozyme activity, neutrophil reactive oxygen species (ROS) production, and total plasma protein levels did not significantly differ between treatment groups; however, plasma cholesterol increased significantly in fish fed GM, AC, NR, and C-HF compared to C-LF (I). A significant increase in plasma glucose and triglyceride was observed in GM and NR treatments, while glucose levels were significantly higher in C-HF compared to C-LF. Moreover, the immunostimulant-supplemented diets exhibited significantly lower cortisol levels compared to controls C-LF (at 0.5 h) and C-HF (at 1 h) post stress, respectively (II). According to our findings, FM substitution did not modulate the innate immune response but was associated with reduced levels of cholesterol. Dietary immunostimulants were not effective enough to boost the immune response, but we believe they might be helpful to trigger metabolic advantages during stressful handling events on fish farms.     

Development of a modified cortisol extraction procedure for intermediately sized fish not amenable to whole-body or plasma extraction methods

The corticosteroid hormone cortisol is the central mediator of the teleost stress response. Therefore, the accurate quantification of cortisol in teleost fishes is a vital tool for addressing fundamental questions about an animal’s physiological response to environmental stressors. Conventional steroid extraction methods using plasma or whole-body homogenates, however, are inefficient within an intermediate size range of fish that are too small for phlebotomy and too large for whole-body steroid extractions. To assess the potential effects of hatchery-induced stress on survival of fingerling hatchery-reared Spotted Seatrout (Cynoscion nebulosus), we developed a novel extraction procedure for measuring cortisol in intermediately sized fish (50–100 mm in length) that are not amenable to standard cortisol extraction methods. By excising a standardized portion of the caudal peduncle, this tissue extraction procedure allows for a small portion of a larger fish to be sampled for cortisol, while minimizing the potential interference from lipids that may be extracted using whole-body homogenization procedures. Assay precision was comparable to published plasma and whole-body extraction procedures, and cortisol quantification over a wide range of sample dilutions displayed parallelism versus assay standards. Intra-assay  %CV was 8.54 %, and average recovery of spiked samples was 102 %. Also, tissue cortisol levels quantified using this method increase 30 min after handling stress and are significantly correlated with blood values. We conclude that this modified cortisol extraction procedure provides an excellent alternative to plasma and whole-body extraction procedures for intermediately sized fish, and will facilitate the efficient assessment of cortisol in a variety of situations ranging from basic laboratory research to industrial and field-based environmental health applications.     

Effects of hormonal manipulation on stress responses in male and female broodstocks of pikeperch Sander lucioperca

This study was designed to determine the effects of hormonal manipulation on stress responses in female and male pikeperch. Two-year-old cultured female and male broodstocks with an average weight of 337.4 ± 20.1 (mean ± SE; n = 16) and 318.7 ± 15.1 g (n = 16), respectively, were randomly allocated into four hormonal treatments each containing 4 fish. Two sexual groups of 16 fish for each gender were considered. Sexually mature male and female pikeperch were injected with either physiological saline solution (as control group), common carp pituitary extract (CPE), human chorionic gonadotropin (hCG) and or luteinizing hormone-releasing hormone analog (LHRHa₂). The blood samples were taken before hormonal injection and after ovulation and spermiation. Then the plasma levels of stress indices (cortisol, glucose, and lactate) were determined. The results showed that all CPE-, HCG-, and LHRHa_2- injected males produced sperm. In females treated with CPE and hCG, three of four ovulated, but none of LHRHa_2- and saline-injected fish spawned. Significant changes in cortisol, glucose, and lactate levels were observed among the females injected with different hormones. Plasma cortisol and glucose levels increased significantly in males injected with CPE and females injected with hCG, but no significant change was observed in lactate levels before and after hormonal induction. Comparison of two sexes revealed significant differences in glucose levels for females in some groups before injection, while CPE-injected sexes showed significant changes in cortisol and lactate concentrations. The results indicated that the induction of ovulation or spermiation stimulated stress responses especially in female pikeperch, and therefore, all the procedures should be made to minimize the disturbance during the artificial spawning.     

Effects of dietary green tea (Camellia sinensis L) on acute stress responses in sturgeon hybrid (Huso huso ♂ × Acipenser ruthenus ♀)

In aquaculture practices, fish commonly confront environmental alterations and unavoidable stresses. Thus, any substance that could attenuate stress responses could be potentially used in aquaculture. A total number of 90 hybrid sturgeons (Huso huso ♂ × Acipenser ruthenus ♀) with an average weight of 212.6 ± 0.7 g were fed experimental diets containing 0, 50 and 100 mg kg^?1 green tea (Camellia sinensis L) for 6 weeks. Subsequently 15 fish from each treatment were exposed to air for 5 min prior to transfer to the recovery tank. Followed by blood sampling after 1, 2, 5 and 12 h, Serum cortisol increased in all treatments, but the intensity was significantly lower in fish fed green tea compared to the control group. Similarly, the green tea diet attenuated the increase in serum levels of glucose and lactate in comparison to the control. Our results showed that green tea could be added to fish diet to attenuate stress responses in Sturgeon hybrid (Huso huso ♂ × Acipenser ruthenus ♀).