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.     

Alternative matrices for cortisol measurement in fish

Plasma cortisol is the most commonly used indicator of stress in fish but, as the blood sampling procedure itself can be a source of stress, it would be helpful to measure cortisol using less invasive matrices. It is also necessary to find alternative matrices as stress indicators in dead fish in which blood sampling is impossible. In the present study, we investigated transport stress in three aquaculture species, European sea bass (Dicentrarchus labrax L.), common carp (Cyprinus carpio L.) and rainbow trout (Oncorhynchus mykiss Walbaum), by cortisol determination (radioimmunoassay) in plasma and other matrices (skin mucus, gut content, lateral muscle and caudal fin). Cortisol significantly increased after transport in all species and matrices, except in the sea bass gut content, where it remained unchanged. The three species responded to transport stress by producing different cortisol levels. In conclusion, the significant correlation found between plasma cortisol and most of the other matrices opens up the possibility of using them to evaluate stress in fish: mucus sampling is a less invasive method than blood sampling, and in addition to muscle and fin sampling, it can be used in postmortem fish.     

Daily rhythms of digestive physiology, metabolism and behaviour in the European eel (Anguilla anguilla)

Circadian rhythms are common in a wide variety of physiological processes. Our knowledge of biological rhythms in fish can be exploited to improve some key aspects of aquaculture such as feed optimization and fish welfare. The aim of this paper was to investigate daily rhythms in behavioural and physiological variables of the European eel. To this end, we first analysed the daily rhythms in locomotor activity measured by means of infrared photocells and feeding activity by means of self-feeders. The eels were then submitted to scheduled feeding (single meals once a day at a fixed time), and the levels of cortisol, glucose and digestive enzymes were analysed. Locomotor activity was predominantly nocturnal (63?% of daily activity occurring at night) with peaks located around light transitions, whereas feeding activity was less clearly defined. Cortisol and glucose showed daily rhythms, with acrophases located in the middle and at the beginning of the light phase, respectively, which should be carefully considered when using these parameters as welfare indicators. Significant variations were found during the day in acid protease levels, which increased in anticipation of mealtime. Taken together, these results describe for the first time a picture of the circadian physiology of the European eel, providing hints to improve feeding methods and welfare assessment.     

Effects of fasting on metabolic and immunological responses of common carp (Cyprinus carpio) to a further acute stress

Common carp were fasted for 0, 3 and 14 days prior to exposure to a 2‐hr crowding stress. Serum cortisol, glucose, lactate, _{T4, T3}, serum alternative complement (ACH50), lysozyme, total immunoglobulin (Ig) and blood WBC were determined before and 0, 6 and 24 hr after the crowding stress. Three days of fasting led to a significant decrease in serum levels of lactate and thyroid hormones. Fourteen days of fasting led to initiation of physiological process to maintain serum glucose by increase in serum cortisol level and lactate utilization. These fish had lower levels of thyroid hormones suggesting suppression of basal metabolism. The fish also had lower WBC and serum lysozyme and total Ig levels indicating immunosuppression. After the crowding stress, the fish fasted for 3 days had significantly lower lactate, thyroid hormones and total Ig levels. The fish fasted for 14 days had higher cortisol but lower glucose and thyroid hormones levels suggesting impairment in metabolic responses to the crowding stress. Immunological and health indicators including WBC, and ACH50, lysozyme and total Ig of these fish were significantly lower than the other groups after the crowding stress. Overall, the results show that a 14‐day fasting leads to stress responses, reduction in metabolism and immunosuppression. Moreover, 14 days of fasting lead to greater cortisol response to a further crowding stress, which induces marked immunosuppression and fallen welfare.     

Modelling the effects of capture and sea lice [Lepeophtheirus salmonis (Krøyer)] infestation on the cortisol stress response in trout

Measuring baseline cortisol in wild salmonids is problematic because of the stress induced by most capture methods. The cortisol response to different periods of confinement stress was modelled in a laboratory population of rainbow trout [Oncorhynchus mykiss (Walbaum)]. A quadratic model was used to predict baseline (precapture) cortisol levels in these fish. A similar quadratic response pattern of cortisol was observed in wild sea trout (Salmo trutta L.). These were captured in timed sequences on four dates by seine netting in Clew Bay, west coast of Ireland. The estimated precapture cortisol for these fish indicated higher levels soon after migration to sea and a positive correlation with the level of infestation by Lepeophtheirus salmonis (Krøyer). Curvilinearity of the quadratic model was correlated with levels of lice infestation, indicating an enhanced cortisol response to confinement stress during capture when parasitic abundance was high. Estimated precapture cortisol for individual sea trout was positively correlated with numbers of lice although the linear regression had low predictive power. The method provides, for the first time, a means to estimate baseline cortisol levels in wild salmonids, and addresses one of the most difficult problems in working with wild fish. Keywords: Cortisol, trout, Lepeophtheirus salmonis (Krøyer), capture, quadratic model     

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。     

Stress responses of Atlantic salmon (Salmo salar L.) elicited by water level reduction in rearing tanks

The Atlantic salmon (Salmo salar) stress response was examined by measuring plasma cortisol, glucose and chloride in fish after water level reduction within rearing tanks. Maximum plasma cortisol levels (366±43 (SD) nM and 534±280 nM for Groups 1 and 2, respectively) were observed 20 min after application of the stressor. Cortisol levels were down to control levels 24 h later. The pattern for cortisol changes observed within these two groups were comparable. Plasma glucose and chloride concentrations did not change significantly in either of the experimental groups. The fish in group 2 were then repeatedly subjected to the same stressor every third day. After the fifth exposure to the stressor, blood was obtained from fish at times corresponding to sampling after the first exposure. Their maximum plasma cortisol level only reached 223 (±96) nM, and was down to prestress levels within 2 h. Plasma glucose and chloride concentrations did not change significantly in this second experiment. This challenge test revealed an acute primary response in Atlantic salmon without any apparent harmful secondary responses, that may thus serve as a standardized reference stressor using other fish groups under comparable conditions.     

Moderate stocking density does not influence the behavioural and physiological responses of rainbow trout (Oncorhynchus mykiss) in organic aquaculture

Welfare in farmed fish got particular attention during the last decades from both governmental and public sides. In aquaculture context, welfare concerns are mainly related to handling procedures, water quality and stoking densities. In Europe, authorities had to clarify the threshold limits of stocking densities to maintain fish good welfare, including for organics aquaculture through the EC regulation 710/2009. However, effects of stocking density on fish welfare are complex and sometimes contradictory. Moreover, there is a lack of knowledge about the impact of density on fish welfare in organic aquaculture. Thus, the aim of the study is to asses welfare state of rainbow trout (Oncorhynchus mykiss) at two initial stocking densities (low density, LD: 12 kg/m³ and high density, HD: 17 kg/m³) fed using organic feed by combining the monitoring of growth performances, behaviour (swimming activity) and physiological indicators (i.e. cortisol, glucose, lactate, hematocrit, red blood cellule count and lysozyme). At the end of experiment, the stocking density reached 21 kg/m³ and 30 kg/m³ for the LD and HD respectively. Overall, growth performances, swimming activity and level of physiological indicators of stress and welfare were similar between HD and LD over the experiment duration. To conclude, we observed no alteration of fish welfare between the two stocking densities monitored. This study suggests that a final stocking density of 30 kg/m³ can be considered for organic aquaculture of rainbow trout respecting welfare.     

Stressing fish in Recirculating Aquaculture Systems (RAS): does stress induced in one group of fish affect the feeding motivation of other fish sharing the same RAS?

As a consequence of water re‐use and high stocking densities, Recirculating Aquaculture Systems (RAS) may lead to an accumulation of substances released by the fish into the water, e.g. cortisol and alarm pheromones. This study investigated the effect of stressing fish on the feeding motivation of other fish not subjected to stress but sharing the same water of stressed fish. Two identical RAS were used (operated at 30 L kg feed^?1 day^?1) and contained grouped (stressed fish) and individually (receiving water from stressed fish) housed Nile tilapia. A stress test was applied in grouped housed fish on days 17 and 55. Feeding behaviour (intake and latency) was recorded in the individually housed fish 3 days before, during and 3 days after the stress test. The results showed no effect on feeding behaviour in fish receiving the water from stressed fish. These results could be a consequence of insufficient cortisol/alarm cues' release by the stressed fish into the water or inactivity of such substances, either due to a trapping effect of humic acids or due to degradation in the nitrification and denitrification processes. Future research is needed to clarify how these processes may affect the water concentration of cortisol and alarm pheromones and should be extended by measuring other behavioural and physiological traits.     

Selection for improved stress tolerance in rainbow trout (Oncorhynchus mykiss) leads to reduced feed waste

The magnitude of the cortisol response to a standardised stressor is a heritable trait in salmonid fishes, and selection for stress responsiveness induces differences in both behaviour and neuroendocrine function. For instance, in laboratory studies, fish with a high cortisol response show a greater propensity for stress induced anorexia. Some authors have, however, commented that controlled studies encompassing relatively small groups of fish hold little or no relevance to practical aquaculture. This notion may be flawed, since understanding the mechanisms behind the behaviour of individuals is a proviso to predict behaviour in groups even with the caveat that some behaviors may be modified by group size. As an example, optimal feeding regimes should be easier to predict in a population consisting of individuals whose appetite is relatively less affected by external stressors. In a fluctuating and potentially stressful environment, such a population should also generate less feed waste, if kept on fixed rations. In the present experiment, we tested this hypothesis by monitoring feed waste and feed conversion efficiency in lines of rainbow trout selected for a low (LR) or high (HR) cortisol response to stress. The study was carried out after fish had been transported between rearing sites in the United Kingdom and Norway. There was significantly more feed waste from rearing units containing HR fish, and these fish also showed lower feed efficiency (growth per unit feed consumed). The difference in feed waste became more apparent with increasing time after transport, as rations increased. Simultaneously, size was more variable and growth was slower in HR rearing units. These results suggest that there are several potential benefits of selection for low stress responsiveness in aquaculture.     

Consistency of stress response to repeated handling in the gilthead sea bream Sparus aurata Linnaeus, 1758

Gilthead sea bream Sparus aurata Linnaeus, 1758 were subjected to repeated stresses, namely confinement and handling once a month for 5 consecutive months. Plasma cortisol concentration was determined as a primary stress response, and plasma glucose and osmolality, serum agglutination and growth rate were analysed as indicators of the secondary and tertiary effects of stress. The results of the mean values after the five trials showed that most indicators followed a consistent pattern with time. Cortisol and growth rate were positively correlated, and cortisol and osmolality were negatively correlated after stress. Analysis of the individual responses throughout all 5 months showed that 6–20% of the fish displayed a consistently high or low response depending on the parameter analysed.     

Stress hormone responses in ayu Plecoglossus altivelis in reaction to different catching methods: comparisons between electrofishing and cast netting

It is important to select the most appropriate method for fish sampling in streams and rivers; the sampling efficiency as well as any negative effect of the method of fish sampling on fishes such as stress, injury, and mortality must be considered. This study aimed to investigate stress hormone responses in ayu Plecoglossus altivelis in reaction to direct current (DC) electrofishing, alternating current (AC) electrofishing, and cast netting in artificial streams. The mortality, injury rate, and catch efficiency of these catching methods were also compared. It was found that immediately after capture, fish caught using a cast net exhibited significantly higher cortisol levels than did control fish that were caught rapidly by a hand net. Cortisol levels did not differ between electroshocked fish and the controls. Time required to catch 12 fish was shorter when DC was used (20 s) than when AC was used (45 s). The time required to catch fish greatly increased when a cast net was used (840 s). Cortisol levels in DC electroshocked and control treatment groups were at resting levels 24 and 48 h after capture. However, higher cortisol levels were found in fish captured using a cast net at both these time points. Cortisol levels in AC electroshocked fish returned to lower levels at 24 h, but increased again at 48 h. Furthermore, 48 h following capture, the fish caught by AC electrofishing exhibited higher mortality (7.5 %) than those captured by other methods (0 %). Spinal injury was also detected in one of the fish in this group. Considering these findings, DC electrofishing is the most effective and least damaging method for collecting P. altivelis in streams with respect to stress, physical damage, and efficiency.     

Anaesthesia of farmed fish: implications for welfare

During their life cycle as farmed animals, there are several situations in which fish are subjected to handling and confinement. Netting, weighing, sorting, vaccination, transport and, at the end, slaughter are frequent events under farming conditions. As research subjects, fish may also undergo surgical procedures that range from tagging, sampling and small incisions to invasive procedures. In these situations, treatment with anaesthetic agents may be necessary in order to ensure the welfare of the fish. The main objective of this paper is to review our knowledge of the effects of anaesthetic agents in farmed fish and their possible implications for welfare. As wide variations in response to anaesthesia have been observed both between and within species, special attention has been paid to the importance of secondary factors such as body weight, water temperature and acute stress. In this review, we have limited ourselves to the anaesthetic agents such as benzocaine, metacaine (MS-222), metomidate hydrochloride, isoeugenol, 2-phenoxyethanol and quinaldine. Anaesthetic protocols of fish usually refer to one single agent, whereas protocols of human and veterinary medicine cover combinations of several drugs, each contributing to the effects needed in the anaesthesia. As stress prior to anaesthesia may result in abnormal reactions, pre-anaesthetic sedation is regularly used in order to reduce or avoid stress and is an integral part of the veterinary protocols of higher vertebrates. Furthermore, the anaesthetic agents that are used in order to obtain general anaesthesia are combined with analgesic agents that target nociception. The increased use of such combinations in fish is therefore included as a special section. Anaesthetic agents are widely used to avoid stress during various farming procedures. While several studies report that anaesthetics are effective in reducing the stress associated with confinement and handling, there are indications that anaesthesia may in itself induce a stress response, measured by elevated levels of cortisol. MS-222 has been reported to elicit high cortisol release rates immediately following exposure, while benzocaine causes a bimodal response. Metomidate has an inhibitory effect on cortisol in fish and seems to induce the lowest release of cortisol of the agents reported in the literature. Compared to what is observed following severe stressors such as handling and confinement, the amount of cortisol released in response to anaesthesia appears to be low but may represent an extra load under otherwise stressful circumstances. Furthermore, anaesthetics may cause secondary adverse reactions such as acidosis and osmotic stress due to respiratory arrest and insufficient exchange of gas and ions between the blood and the water. All in all, anaesthetics may reduce stress and thereby improve welfare but can also have unwanted side effects that reduce the welfare of the fish and should therefore always be used with caution. Finally, on the basis of the data reported in the literature and our own experience, we recommend that anaesthetic protocols should always be tested on a few fish under prevailing conditions in order to ensure an adequate depth of anaesthesia. This recommendation applies whether a single agent or a combination of agents is used, although it appears that protocols comprising combinations of agents provide wider safety margins. The analgesic effects of currently used agents, in spite of their proven local effects, are currently being debated as the agents are administrated to fish via inhalation rather than locally at the target site. We therefore recommend that all protocols of procedures requiring general anaesthesia should be complemented by administration of agents with analgesic effect at the site of tissue trauma.     

捕食胁迫对“四大家鱼”幼鱼生理反应的影响

本研究选取乌鳢(Channa argus)和南方大口鲶(Silurus soldatovi meridionalis)为捕食者,青鱼(Mylopharyngodon piceus)、草鱼(Ctenopharyngodon idellus)、鲢(Hypophthalmichthys molitrix)、鳙(Aristichthys nobilis)幼鱼为猎物鱼,比较了在无捕食(空白对照)、低捕食(隔网胁迫)和高捕食(直接胁迫)压力下,胁迫0、7、14 d后,“四大家鱼”幼鱼血清皮质醇(COR)水平和血液生化指标的变化。结果显示,不同捕食胁迫水平下,“四大家鱼”幼鱼的血液生化指标和COR水平变化程度不同,但变化趋势一致。“四大家鱼”幼鱼的COR水平随着捕食胁迫程度和胁迫时长的增加显著升高,表现为无捕食组<低捕食组<高捕食组,0 d<7 d<14 d。在血液生化指标中,血清蛋白浓度和总胆固醇(CHO)浓度较为稳定,各组相比无显著变化;血糖(GLU)浓度和碱性磷酸酶(ALK)在捕食胁迫下升高,甘油三酯(TG)则相反,呈下降趋势。研究表明,“四大家鱼”幼鱼会根据捕食风险来增强自身生存能力的方式及调整生理反应。捕食胁迫处理后,“四大家鱼”幼鱼均产生了应激反应,与隔网捕食相比,直接捕食对鱼体生理反应影响更为显著,且随着胁迫时长的增加应激程度也随之增加。各检测指标中,血清蛋白和CHO可能不是捕食胁迫下鱼类应激的敏感指标;COR和GLU的变化最为显著,这可能是为了弥补应激期间机体对能量需求的增大。     

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.     

Ontogeny of the stress response in chinook salmon, Oncorhynchus tshawytscha *

Whole body concentrations of cortisol were determined via radioimmunoassay in chinook salmon, Onchorynchus tshawytscha, during early development in both stressed and non-stressed fish to determine when the corticosteroidogenic stress response first appeared. Progeny from both pooled and individual females were examined to determine if differences existed in offspring from different females. Levels of cortisol were low in eyed eggs, increased at hatch, decreased 2 weeks later and then remained constant thereafter. Differences in cortisol between stressed and control fish were found 1 week after hatch and persisted for the remainder of the study. The magnitude of the stress response, or relative amount of cortisol produced, generally increased from the time when it was first detected, but a decrease in the ability to elicit cortisol was seen 4 weeks after hatching. Cortisol content of separate progeny from two individual females showed a similar pattern to that seen in pooled eggs. Our results indicate that chinook salmon are capable of producing cortisol following a stressful event approximately 1 week after the time of hatching. The decrease in endogenous cortisol content seen 2 weeks after hatching, and the decrease in the magnitude of the stress response seen 4 weeks after hatching may be comparable to developmental events documented in mammals where corticosteroid synthesis is inhibited to neutralize possible detrimental effects of these hormones during critical periods of development. This revised version was published online in August 2006 with corrections to the Cover Date.     

The effect of allostatic load on hypothalamic–pituitary–interrenal (HPI) axis before and after secondary vaccination in Atlantic salmon postsmolts (Salmo salar L.)

The experiment consisted of three experimental groups: (1) “vaccine and stress”, (2) “stress and vaccine” and (3) control. All groups have previously been vaccinated 6 months prior to the start of the experiment. At the start of the experiment, the “vaccine and stress” group was vaccinated with Pentium Forte Plus for the second time (25.02.2008) and then given a daily stressor (confinement stressor 267 kg m-3 in 15 min) for a period of 4 weeks. The “stress and vaccine” group was given a similar daily stressor for 4 weeks and then vaccinated for the second time. The control group was neither stressed nor vaccinated a second time. The results indicates that fish in the “stress and vaccine” group may have entered an allostatic overload type 2 due to oversensitivity to ACTH, a reduced efficient negative feedback system with elevated baseline levels of plasma cortisol and reduced immune response with pronounced effects on the well-being of the animal. The “vaccine and stress” group may likewise have entered an allostatic overload type 1 response, with oversensitivity to ACTH and transient reduced efficient negative feedback system. This study shows that if plasma cortisol becomes elevated prior to vaccination, it could perhaps instigate an allostatic overload type 2 with dire consequences on animal welfare. To reduce the risk of compromising the animal welfare during commercial vaccination of salmon, one propose to grade the fish minimum a week prior to vaccination or grade simultaneously with vaccination. This could reduce the overall allostatic load during handling and vaccination and secure a healthy fish with intact immune response and improved animal welfare.     

Effect of some repetitive factors on turbot stress response

The stress response of the turbot, Scopthalmus maximus (L.), to repetitive factors including netting, air exposure, blood sampling and hand-stripping, were tested using two different tank sizes as well as two blood sampling techniques. Exposure of juvenile fish to air for 1-4 min had no immediate effect on plasma cortisol concentrations or haematocrit values. Similarly, the serial netting of immature fish from tanks did not significantly modify plasma cortisol concentrations, haematocrit or osmolarity values. Hand-stripping of mature males was more disturbing than air exposure. The cumulative effect on plasma cortisol levels and osmolarity of stress factors such as netting, air exposure, blood sampling and stripping applied simultaneously to mature males in a 16 m3 tank and repeated twice daily for several days was recorded. The level of cortisol increased from 5 ng ml-1 to 300 ng ml-1 after 10 days of treatment, while an osmoregulatory imbalance and fish death were observed. Moreover, adaptation of fish to smaller tanks seemed to improve the increased plasma cortisol levels and death rate. Reduction in the number of stress factors applied greatly decreased both the range of physiological responses and the death rate. In order to avoid a cumulative stress response, handling of fish should therefore be reduced to a minimum.     

Fish stress and mortality can be predicted using reflex impairment

Fish stress and mortality can be significant problems in both capture and culture operations. In addition to inexpensive and real-time metrics, ones that are simple to use are also desirable for measuring fish stress and predicting mortality. Current methods to define stress rely on expensive, laboratory-based measurements of changes in fish pathology such as disease, necropsy and histology, in physiology such as plasma cortisol, lactate, glucose and ions and in complex behaviour determined from swimming, feeding and predator evasion. All of these methods are often not rigorously linked to fitness outcomes. An alternative is to observe reflex impairment as a direct sign of stress which can be easily and rapidly measured in free swimming or restrained fish responding to peripheral stimuli such as gravity, light, sound and touch. Reflex impairment is correlated with stress and mortality outcomes, eliminating the need for prolonged holding or monitoring of fish. A few examples of reflexes that may be impaired include orientation, startle responses, fin erection, body flex upon restraint, operculum and mouth clamping or gaping, gag response and vestibular–ocular response. Reflex impairment combines the effects of stressors and their interactions and is not dependent on fish size, motivation states and acclimation which make it a consistent sign of stress across a wide range of stressor types and fish ages. Use of reflex impairment to measure stress and predict mortality would significantly improve monitoring of fish health and welfare in many types of field operations such as commercial and recreational fishing, aquaculture, live transport, stock enhancement and tagging.     

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.