The effect of stress and exercise on post-mortem biochemistry of Atlantic salmon and rainbow trout

Freshwater Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss responded similarly to increase in water flow (exercise), reduction in holding tank water level (stress), or 30 min chasing with water level reduction (stress and exercise). Stress generally resulted in elevated plasma cortisol, above the control. Fish responded to stress and exercise combined, with elevated lactate and [H⁺] which was sometimes associated with elevated plasma cortisol. These changes were combined with a depletion of the muscle adenylate pool. Post-mortem, this resulted in an increase in the rate of onset of rigor, and a higher and sometimes sustained muscle proton load. Both species produced predominantly inosine as opposed to hypoxanthine, for up to 72 h of ice storage. This study shows that the physiological disruption in Atlantic salmon and rainbow trout caused by simulated harvest conditions of stress and exercise, results in mostly transient changes in post-mortem muscle biochemistry. These changes lead to an earlier onset and resolution of rigor, and lower post-mortem muscle pH in comparison to the control.     

Lack of glucose and hsp70 responses in haddock Melanogrammus aeglefinus (L.) subjected to handling and heat shock

Juvenile haddock Melanogrammus aeglefinus ( c. 39 g) were exposed to either a handling stressor (1 min out of water) or heat shock (increase from 10 to 15° C for 1 h), and plasma cortisol, plasma glucose and gill hsp70 levels were determined before, and at 1, 3, 6, 12, 24 and 48 h post-stress. The pattern of cortisol increase was similar following both stressors, with levels increasing by 25-fold at 1 h post-stress, but returning to pre-stress levels (2–5 ng ml⁻¹) by 3 h. In contrast, neither handling nor heat shock caused an increase in plasma glucose levels. Although gill hsp70 was detected, presumably constitutive levels, in both control and heat shocked groups, there were not significant changes in gill hsp70 levels after exposure to heat shock. The lack of glucose and hsp70 responses to these typical stressors is consistent with previous studies on Atlantic cod Gadus morhua , and suggests that the stress physiology of Gadidae differs from the 'typical' teleost.     

Acid–base and respiratory effects of confinement in Atlantic salmon affected with amoebic gill disease

Amoebic gill disease (AGD) in cultured salmonids causes severe multifocal hyperplastic lesions in the gills with the potential to influence respiratory and acid–base physiology. Atlantic salmon Salmo salar affected with AGD were surgically implanted with dorsal aortic catheters and, following recovery, were confined for 5 min ( n  = 16) or left undisturbed ( n  = 8). Confinement caused an acute extracellular acidosis that was corrected in 6 h amongst surviving fish. There was a gradual increase in plasma lactate concentrations that peaked at 1 h post-confinement then declined by 9 h recovery. In a second experiment, AGD-affected fish were confined then recovered either in a tank of static water ( n  = 9) or while being forced to swim at 1·5 body lengths s⁻¹ ( n  = 6). There was no significant difference between fish recovered by swimming and those in static water in terms of recovery of the acute extracellular acidosis and lactate accumulations coincident with exhaustive exercise. Confinement severely compromised the survival of AGD-affected Atlantic salmon, although survivors appeared to recover similarly to other studies. Forced swimming of AGD-affected Atlantic salmon following confinement did not facilitate recovery and is unlikely to be a useful strategy for mitigating the effects of stressful episodes such as crowding and fish movement and commercial handling.     

Changes in free and total plasma cortisol levels in juvenile haddock (Melanogrammus aeglefinus) exposed to long-term handling stress

We measured changes in free and total plasma cortisol levels, plasma glucose, gill hsp70 levels, and growth in haddock (Melanogrammus aeglefinus) subjected to a long-term handling stress (15 s out of water, each day, for 4 weeks), and the effect of this long-term stress on the ability of haddock to respond to an acute stressor. The acute stressor was a single handling stress, and fish were sampled at 1, 6, and 12 h post-stress. During the long-term stress study, free and total plasma cortisol levels increased significantly (10-fold) in the stressed group after the second week. However, the percentage of free cortisol was already significantly elevated by the first week (control 17%, stressed 55%), and remained high during the second week (control 35% and stressed 65%). After 3 and 4 weeks of handling, both free and total cortisol declined in stressed fish to levels that were not significantly different from pre-stress values. Control fish grew significantly more than stressed fish (by 32% and 18%, respectively) over the 4 week study, and condition factor only increased in control fish. Although fish from the control group showed elevated total plasma cortisol levels (to 47 ng mL(-1)) 1 h after the acute stress, and the levels in stressed fish were comparable to those for the control fish, no significant increase in plasma cortisol was measured in the group subjected to the long-term stress. Free plasma cortisol levels did not increase significantly in either group following the acute stress. However, free plasma cortisol levels were significantly higher in long-term stress group, as compared with the control group, at 6 h post-stress. Plasma glucose and gill hsp70 levels were not altered by either the long-term stress or acute stressor. Our data indicate that cortisol (free and total), but not glucose or hsp70, appears to be adequate to assess short- and long-term stress in haddock.     

Swimming performance and blood chemistry in Atlantic salmon spawners exposed to acid river water with elevated aluminium concentrations

Exposure of sexually mature pre-spawning Atlantic salmon Salmo salar to Fossbekk water (pH 5·2) for 7 days led to a significant reduction in critical swimming speed (U-crit) in females but not in males. Exposure to Fossbekk water +A1 (as AlCl₃) for 24 h led to a significant reduction in U-crit in both males and females. In contrast to fish exposed for 7 days to Fossbekk water, fish exposed to Fossbekk+A1 had accumulated much more aluminium and mucus on their gills. Losses of plasma ions were similar in both groups exposed to acid water. Blood glucose was twice as high in fish exposed to Fossbekk water for 7 days compared with fish exposed to Fossbekk+A1 for 24 h. Plasma cortisol was still elevated compared with controls after exposure to Fossbekk water for 7 days. Possible mechanisms for the observed decrease in U-crit at the different exposures are discussed.     

Haematology of juvenile Acipenser oxyrinchus and Acipenser brevirostrum at rest and following forced activity

In vivo experiments were conducted to examine the haematology of juveniles from two relic bony fishes, Atlantic sturgeon Acipenser oxyrhinchus and shortnose sturgeon Acipenser brevirostrum . Oxygen transport characteristics (haematocrit, haemoglobin and mean erythrocytic haemoglobin concentration), ionic composition (Na⁺, Cl⁻, K⁺ and osmolality), metabolite concentration (lactate, cortisol and glucose) and protein content in blood were measured or calculated at rest and during recovery from forced activity. Under resting conditions, plasma osmolality and concentrations of Na⁺, Cl⁻, lactate, cortisol and total protein were significantly different between Atlantic and shortnose sturgeon. All other resting variables were not different between species. Following forced activity, plasma lactate levels were significantly higher in both species than at rest. Plasma cortisol levels in both species were only significantly higher 1 h following forced activity compared to resting values. Plasma lactate levels were significantly higher in Atlantic sturgeon than in shortnose sturgeon, but these levels returned to resting levels by 1 h in both species. Cortisol increases were greater in shortnose sturgeon than in Atlantic sturgeon. In general, oxygen transport characteristics, blood glucose, plasma protein and plasma osmolality were not altered by forced activity in either sturgeon species. Overall, both species had reduced responses ( i.e . the magnitude of changes in measured variables) to forced activity compared with teleosts.     

Physiological stress responses to confinement in diploid and triploid Atlantic salmon

Confinement of juvenile Atlantic salmon Salmo salar from four different populations (all–female diploids, all–female triploids, mixed sex diploids and mixed sex triploids) either before (FW parr) or after (SW smolts) transfer to sea elevated plasma cortisol and plasma lactate from control levels irrespective of ploidy status. Both before and after confinement, plasma cortisol levels in SW smolts (6–174 ng ml^–1) were higher than those in FW parr (4–58 ng ml^–1), which possibly reflected the physiological challenge of acclimation to SW. Mixed sex populations of SW smolts had higher cortisol levels than all–female populations. The duration of confinement (1, 3 or 6 h) affected the magnitude of the plasma cortisol and lactate responses in SW smolts. Plasma cortisol levels in diploid and triploid SW smolts subjected to 2 h of confinement decreased to pre–stress levels within 6 h post–confinement. Plasma lactate levels were not significantly different from pre–stress levels 48 h after confinement. As no difference exists in primary and secondary stress responses of Atlantic salmon of differing ploidy status, it is unlikely that differences in mortality rates between diploid and triploid populations under commercial conditions can be attributed to differences in their physiological responses to periods of stress lasting up to 6 h.     

Changes in hydrolytic enzyme activities of naïve Atlantic salmon Salmo salar skin mucus due to infection with the salmon louse Lepeophtheirus salmonis and cortisol implantation

The changes in the activities of mucus hydrolytic enzymes and plasma cortisol levels were examined following infection of Atlantic salmon Salmo salar with the salmon louse Lepeophtheirus salmonis and these changes were compared with those resulting from elevated plasma cortisol. Salmon were infected at high (Trial 1; 178 +/- 67) and low (Trial 2; 20 +/- 13) numbers of lice per fish and the activities of proteases, alkaline phosphatase, esterase and lysozyme in the mucus, as well as plasma cortisol levels were determined. At both levels of infection, there were significant increases of protease activity over time (1-way K-WANOVA; Trial 1, p = 0.004; Trial 2, p < 0.001). On several sampling days, generally on later days in the infections, the mucus protease activities of infected fish were significantly higher than control fish (Student's t-tests; p < 0.05). In addition, zymography experiments demonstrated bands of proteases at 17 to 22 kDa in the mucus of infected salmon that were absent in the mucus from non-infected fish and absent in the plasma of salmon. The intensity of these protease bands increased in the mucus over the course of both infections. However, plasma cortisol levels were elevated only in the heavily infected fish from the first trial. At high infection levels (Trial 1), alkaline phosphatase activity was higher in the mucus of infected fish at all days (t-test, p < 0.05). However, at the lower infection level (Trial 2), the mucus alkaline phosphatase activity did not differ significantly between infected and non-infected fish. Esterase and lysozyme activities were very low and did not change with time nor between non-infected and infected salmon in either challenge. Mucus enzyme activities of cortisol-implanted salmon did not change over time, nor were there any differences in activities between cortisol-implanted and control salmon. The present study demonstrates biochemical changes resulting from sea lice infection of Atlantic salmon occurring at the site of host-pathogen interaction, the mucus layer. However, the origin of these enzymes, whether host or pathogen, remains to be determined.     

硬膜外复合全身麻醉对开胸手术患者血流动力学和应激反应的影响研究

目的:研究硬膜外复合全身麻醉对开胸手术患者血流动力学和应激反应的影响。方法:选取120例我院2015年1月到2016年1月收治的开胸手术患者,按照随机数字表法将患者分为研究组和对照组,对照组给予单纯全麻,研究组给予硬膜外复合全身麻醉,比较两组苏醒时间、苏醒期收缩压、舒张压、心率及烦躁情况,并比较两组术前、术后72 h血糖、皮质醇以及肾上腺素水平。结果:两组苏醒时间比较无统计学意义(P0.05),苏醒期研究组收缩压、舒张压及心率、烦躁发生率均较对照组降低(P0.05);两组术前血糖、皮质醇及肾上腺素比较无统计学差异(P0.05);术后72 h,两组血糖、皮质醇及肾上腺素均升高(P0.05);研究组血糖、皮质醇及肾上腺素均较对照组降低(P0.05)。结论:硬膜外复合全身麻醉应用于开胸手术能显著提高患者苏醒期苏醒质量,稳定血流动力学,改善其应激反应。     

Coho salmon haematological, metabolic and acid‐base changes during exercise and recovery in sea water

Cannulated, seawater‐acclimated coho salmon Oncorhynchus kisutch were swum to exhaustion in a seawater tunnel (10° C, mean U _crit 50 cm s⁻¹), resulting in metabolic acidosis and increased plasma electrolyte and cortisol concentrations, which were corrected during a 4 h recovery. Because the swimming and physiological performance data were similar to those of other salmonids, it was concluded that life‐history limitations, besides their exercise capabilities in upwelling zones, probably explain declining coho salmon populations.     

Acute exercise effect on postabsorptive serum leptin.

We postulated that high circulating cortisol levels during intense exercise would lead to increased serum leptin concentrations. Young, lean men ate a small meal and then exercised on a cycle ergometer for 41 min or rested on a control day. Serum leptin concentration was 10% greater during exercise than in the control condition (P < 0.05). Directly after exercise, serum leptin dropped to approximately 10% less than the control level (P < 0.05) but had recovered to the nonexercised level after approximately 2 h of recovery. Rapid exercise effects on circulating leptin were related to changes in hemoconcentration rather than changes in leptin mass. When serum leptin was normalized to serum protein, leptin increased by 10% in the exercise condition compared with control by the end of recovery (P < 0.05). Although exercise increased serum cortisol concentration threefold, there was no relation between differences in cortisol and exercise vs. control differences in normalized leptin. The increased leptin mass after exercise may have been related to greater plasma glucose concentration during recovery after exercise compared with the control condition.     

Recovery of the brown trout, Salmo trutta L., from acute handling stress: a time-course study

The response of the brown trout to a single, short ( c . 1 min) incidence of handling stress was monitored for a period of 1 month post-stress. Significant changes were found in feeding behaviour, in the levels of plasma cortisol, glucose and lactate, in the concentration of circulating lymphocytes and in the degree of epidermal mucification. No changes were detected in the growth rate and coefficient of condition, in the levels of plasma thyroxine, in the concentrations of circulating erythrocytes, neutrophils and thrombocytes or in the thickness of the epidermis. The time-course for recovery of each parameter was examined and from this it was concluded that a minimum of 2 weeks was required for complete recovery and a return to normality.     

Effects of external and surgically implanted dummy radio transmitters on mortality, swimming performance and physiological status of juvenile masu salmon Oncorhynchus masou

The effects of external and surgically implanted radio transmitters on juvenile masu salmon Oncorhynchus masou were examined. External attachment of transmitters significantly decreased the survival of fish and caused a decreased critical swimming speed compared with surgical implantation. Although plasma cortisol, plasma glucose and haematocrit values did not differ significantly among groups, it appeared that the most suitable transmitter attachment method for juvenile O. masou may be surgical implantation.     

Effects of copper on the acute cortisol response and associated physiology in rainbow trout

The aim of this study was to determine the effects of chronic waterborne copper (Cu) exposure on the acute stress-induced cortisol response and associated physiological consequences in rainbow trout (Oncorhynchus mykiss). Trout were exposed to 30 μg Cu/L in moderately hard water (120 mg/L as CaCO(3)) for 40 days, following which time the acute cortisol response was examined with a series of stressors. At 40 days, a 65% increase in Cu was observed in the gill, but no accumulation was observed in the liver, brain or head kidney. Stressors such as air exposure or confinement did not elicit an increase in circulating cortisol levels for Cu-exposed fish, in contrast to controls. However, this inhibitory effect on the acute cortisol response appeared to have few implications on the ability of Cu-exposed fish to maintain ion and carbohydrate homeostasis. For example, plasma Na(+), Ca(2+) and glucose levels as well as hepatic glycogen levels were the same post-stress in control and Cu-exposed fish. Trout were also challenged with exposure to 50% seawater for 48 h, where Cu-exposed trout maintained plasma Na(+), glucose and hepatic glycogen levels. However, Cu-exposed fish experienced decreased plasma K(+) levels throughout the Cu exposure and stress tests. In conclusion, chronic Cu exposure resulted in the abolition of an acute cortisol response post-stress. There was no Cu accumulation in the hypothalamus-pituitary-interrenal axis (HPI axis) suggesting this was not a direct toxic effect of Cu on the cortisol regulatory pathway. However, the lack of an acute cortisol response in Cu-exposed fish did not impair the ability of the fish to maintain ion and carbohydrate homeostasis. This effect on cortisol may be a strategy to reduce costs during the chronic stress of Cu exposure, and not endocrine disruption as a result of toxic injury.     

The effects of stress induced by cortisol administration on the repeatability of swimming performance tests in the European sea bass (Dicentrarchus labrax L.)

The impact of stress induced by cortisol administration on the swimming performance of the European sea bass was tested measuring the fish recovery capability using a respirometer. The recovery test consisted of two exhaustive swimming exercises (U _crit) separated by a recovery period. Thirty-eight fish were subjected to this trial preceded or not (control group) by a cortisol (cortisol group) or saline (placebo) injection. For validation, the classical stress parameters (cortisol, glucose, lysozyme, Hct, Hb and RBCC) were monitored. During the recovery tests, only the cortisol group showed a significant reduction in the recovery capability that was interpreted as a sign of possible physiological impairment. Hct, Hb and RBCC did not show significant inter and intra-group differences, while cortisol and glucose increased as a response to fatigue in all the groups at the end of the trials. No immune (lysozyme) response was observed in control and placebo groups, while a reduction was evidenced in the cortisol group.     

Influence of colostrum intake on piglet survival and immunity

Colostrum intake from birth to 24 h after the onset of parturition (T24) was estimated for 526 piglets from 40 litters. Plasma concentrations of immunoglobulin G (IgG), lactate, glucose and cortisol were determined at T24 for six piglets per litter. Plasma IgG concentration was also assayed at weaning (28 days) on the same piglets. Rectal temperature was measured at T24 on all piglets. Mortality was recorded until weaning and comparisons were made between piglets that died before weaning and those that were still alive at weaning. The piglets that died before weaning had lower birth weight, lower colostrum intake, lower weight gain between birth and T24, and had a lower rectal temperature, higher plasma cortisol concentration and lower plasma IgG and glucose concentrations at T24 than piglets still alive at weaning. In addition, a higher proportion of piglets that died before weaning had difficulty taking their first breath after birth and were affected by splayleg. Considering all piglets, colostrum intake was positively related to rectal temperature and plasma glucose concentration and negatively related to plasma cortisol concentration at T24. Plasma IgG concentration at T24 was explained by colostrum intake, IgG concentration in the ingested colostrum, birth weight and birth rank (P<0.0001). Plasma IgG concentration at weaning was related to plasma IgG concentration at T24 (r=0.54; P<0.0001) and to colostrum intake (r=0.32; P<0.0001). Finally, body weight was explained by colostrum intake, birth weight and age until 6 weeks of age (P<0.0001). These results show that colostrum intake is the main determinant of piglet survival through provision of energy and immune protection and has potential long-term effects on piglet growth and immunity.     

Liver-derived endocrine IGF-I is not critical for activation of skeletal muscle protein synthesis following oral feeding

Background

Like humans, fish can be classified according to their athletic performance. Sustained exercise training of fish can improve growth and physical capacity, and recent results have documented improved disease resistance in exercised Atlantic salmon. In this study we investigated the effects of inherent swimming performance and exercise training on disease resistance in Atlantic salmon. Atlantic salmon were first classified as either poor or good according to their swimming performance in a screening test and then exercise trained for 10 weeks using one of two constant-velocity or two interval-velocity training regimes for comparison against control trained fish (low speed continuously). Disease resistance was assessed by a viral disease challenge test (infectious pancreatic necrosis) and gene expression analyses of the host response in selected organs.

Results

An inherently good swimming performance was associated with improved disease resistance, as good swimmers showed significantly better survival compared to poor swimmers in the viral challenge test. Differences in mortalities between poor and good swimmers were correlated with cardiac mRNA expression of virus responsive genes reflecting the infection status. Although not significant, fish trained at constant-velocity showed a trend towards higher survival than fish trained at either short or long intervals. Finally, only constant training at high intensity had a significant positive effect on fish growth compared to control trained fish.

Conclusions

This is the first evidence suggesting that inherent swimming performance is associated with disease resistance in fish.     

Cortisol and glucose responses after acute stress by net handling in the sparid red porgy previously subjected to crowding stress

In red porgy Pagrus pagrus subjected to 3 weeks of chronic stress by crowding, plasma cortisol remained significantly higher in crowded fish compared to controls. There was no significant effect of crowding on plasma glucose levels. When the crowded fish were subjected to an acute handling the plasma cortisol response was similar to that of the uncrowded fish. No significant differences were found between the groups. The changes in plasma glucose following acute handling were also similar in both crowded and uncrowded fish.     

Could the essential oil of Lippia alba provide a readily available and cost-effective anaesthetic for Nile tilapia (Oreochromis niloticus)?

We report the optimal concentration of the essential oil of Lippia alba (EOLA) for the induction and recovery of anesthesia in Nile tilapia (Oreochromis niloticus) during handling. Groups of 10 juveniles were separately subjected to a concentration of EOLA: 0 (control group), 10, 20, 50, 100, 200, 300, or 500 µL L-1. The plasma cortisol levels were significantly decreased in anesthetized fish 4 h after stress of handling. The plasma glucose levels were significantly increased 1 and 4 h after stress of handling in fish exposed to EOLA. Sensory analysis showed that tasters did not detect differences through a blind test in fillet taste and odor between exposed and non-exposed fish. The ideal concentration to promote the best anesthesia and recovery times and to reduce plasma cortisol 4 h after handling is 500 µL L-1.