Impairment of the stress response in matrinxã juveniles (Brycon amazonicus) exposed to low concentrations of phenol

In this study we measured plasma cortisol, plasma glucose, plasma sodium and potassium, and liver and gill hsp70 levels in juvenile matrinxã (Brycon amazonicus) subjected to a 96 h exposure to phenol (0, 0.2, and 2.0 ppm), and the effect of this exposure on their ability to respond to a subsequent handling stress. Fish were sampled prior to initiation of exposure and 96 h, and at 1, 6, 12, and 24 h post-handling stress. During the 96 h exposure, plasma cortisol and glucose levels remained unchanged in all treatments. While plasma sodium levels were significantly reduced in all groups, plasma potassium levels only decreased in fish exposed to 0 and 0.2 ppm of phenol. Liver hsp70 levels decreased significantly at 96 h in fish exposed to 2.0 ppm of phenol. All groups, except fish exposed to 0.2 ppm of phenol, were able to increase plasma cortisol and glucose levels after handling stress. Fish exposed to 2.0 ppm of phenol showed decreased gill and liver hsp70 levels after the handling stress. Our data suggest that exposure to phenol may compromise the ability of matrinxã to elicit physiological responses to a subsequent stressor.     

Physiological stress effects of continuous- and pulsed-DC electroshock on juvenile bull trout

Juvenile bull trout Salvelinus confluentus exposed to continuous- or pulsed-DC electroshock exhibited rapid elevations in plasma cortisol and glucose, but plasma chloride did not change. In a 1-h experiment using 240 V at 1·4 A of 60-Hz pulsed DC (voltage gradient 0·81 V cm⁻¹), which proved lethal, plasma cortisol and glucose rose significantly within 15 min of a 10-s electroshock. Plasma cortisol reached a peak level of 156 ± 18 ng ml⁻¹ at 45 min and then decreased, whereas plasma glucose reached its highest level of 179 ± 7·5mg dl⁻¹ at 1 h. In a 24-h experiment using lower dosages, plasma cortisol increased from 6·1-16 ng ml⁻¹ to peak levels of 155–161 ng ml⁻¹ in 1 h in response to a 10-s electroshock of continuous (130 V, 0·5 A, 1·45 V cm⁻¹) or pulsed (120 V, 0·5 A, 60 Hz, 0·55 V cm⁻¹) DC. Although plasma concentrations declined thereafter, levels remained above control values at 24 h. Plasma glucose was elevated from 60–65 to 120–134 mg dl⁻¹ after 1h by both electroshock treatments and remained near or above those levels for the 24-h duration. Plasma cortisol and glucose levels were much higher in electroshocked bull trout at 1 h compared with those in fish 1 h after receiving a 30-s handling stressor (cortisol, 90 ± 12 ng ml⁻¹; glucose, 82 ± 6·1 mg dl⁻¹). The results indicate that both continuous and pulsed DC were more stressful to juvenile bull trout than handling and that recovery, at least for pulsed DC, may take longer than 24 h.     

Effects of abrupt cold shock on stress responses and recovery in brown trout exhausted by swimming

To simulate swimming in a trawl, age 3 year brown trout Salmo trutta (L.) were made to swim against a flow of 0·5 m s⁻¹ for 60 min. To simulate cold shock, similar to placing them in a chilling tank, the fish were kept for 10 min in a tank containing ice and water. To simulate the combined stressors, the fish were first made to swim followed by a cold shock. The fish were in a comatose state 10 min after cold shock and combined stressors but conscious after swimming only. All the fish survived until the end of the studied recovery period (maximum 24 h). Cold shock after swimming (combined stressors v . swimming only) did not produce higher blood cortisol, lactate or glucose concentrations 10 min after the treatment. The effect of cold shock, however, was evident in the delayed start of recovery in cortisol and glucose concentrations. All the stress indicators used decreased to the levels for undisturbed fish within 24 h, except in the case of glucose after the combined stressors.     

Responses to handling and confinement stressors in juvenile great sturgeon Huso huso

The effects of acute stressors on physiological responses of juvenile great sturgeon or beluga Huso huso L. were investigated in two experiments. In the first experiment, fish were handled by placing them in containers at either low density (LD, one fish l^?1) or high density (HD, four fish l^?1) for 60 s. Concentrations of plasma cortisol, glucose and lactate were determined from blood collected at 0, 1, 3, 6 and 12 h after application of the stressor. Plasma cortisol concentrations increased after the disturbance in H. huso from both handling treatments, but changes were not significant. Plasma glucose rose significantly by 22·9 and 31·6% in LD and HD handling treatments, respectively, after 3 h. Significant increases in plasma lactate occurred within 1 h in both treatment groups, but that of the HD group was much higher. In the second experiment, fish were held at two different densities, LD (2 kg m^?2 tank bottom surface area) and HD (7 kg m^?2), for 8 weeks and then subjected to an aerial emersion handling stressor in a net for 60 s; blood samples were taken before handling (resting, 0 h) and at 1, 3, 6 and 9 h after handling. Plasma cortisol increased significantly in fish from the HD treatment from 8·8 ± 0·3 to 19·2 ± 2·4 ng ml^?1 (mean ±s.e. ) by 1 h after stress, but post‐handling changes in the LD group were not significant. Significant increases in both plasma glucose and lactate were observed by 1 h in both treatment groups, with peak levels of plasma glucose evident at 3 h [69·4 ± 2·9 and 60·9 ± 1·7 mg dl^?1 (mean ±s.e .) in LD and HD groups, respectively]. Plasma glucose levels were significantly higher in the LD group than in the HD group at 3 and 6 h. Post‐handling haemoglobin content increased by 1 h and white blood cell numbers were reduced by 3 and 6 h in the HD treatment group compared with resting values, but changes in these blood features in the LD group were not significant. Acute handling did not affect haematocrit in either treatment. The results suggest that H. huso is relatively resistant to handling and confinement, and could tolerate normal hatchery practices associated with aquaculture. Because changes in cortisol concentrations were relatively low compared with those in most teleosts, glucose and lactate concentrations may be more useful as stress indicators in juvenile H. huso. This study also demonstrated that prior exposure to a chronic stressor, specifically high stocking density, could alter the physiological response to subsequent acute handling in H. huso.     

Recovery of wild, juvenile brown trout from stress of flow reduction, electrofishing, handling and transfer from river to an indoor simulated stream channel

Stress in wild brown trout Salmo trutta was assessed by sampling blood and measuring the concentrations of plasma cortisol and blood glucose in fish collected by electrofishing and immediately anaesthetized with metomidate. In the River Nidelva, Trondheim, Norway, the resting blood plasma cortisol concentration in the juvenile (0 + year) brown trout was 52 ± 44 nM (mean ± s .d .) in December and 2·3 nM (detection limit) in January. The corresponding blood glucose values were 1·8 ± 0·9 and 1·2 ± 0·2 mM, respectively. After electrofishing, handling and transport to the artificial stream, plasma cortisol and blood glucose levels increased significantly in both experiments. A maximum plasma cortisol level of 239 ± 120 and 71 ± 32 nM and a maximum blood glucose level of 3·9 ± 0·9 and 3·0 ± 0·9 mM were measured in the December and January stream channel experiments after transport, respectively. After introduction to the artificial stream, the blood plasma cortisol level returned to resting values within 24 h in the January stream channel experiment. The blood glucose levels remained at a higher level compared to the reference group throughout the December experiment, while it returned to resting values after 24 h in the January stream channel experiment. The major difference between the December and January experiments was the temperature within the artificial stream, 15–17° C in December and 7–9° C in January. This may have influenced the blood glucose levels. After dewatering of the artificial streambed there was a significant increase in plasma cortisol both in the December and January experiments, and after 24 h the plasma cortisol returned to the resting level in the January experiment. The blood glucose also increased during dewatering, although not significantly.     

Water pH and urinary excretion in silver catfish Rhamdia quelen

The effect of acute exposure to different water pH levels on urinary excretion and plasma ion levels in silver catfish Rhamdia quelen was analysed. Fish were exposed to pH 4·0, 5·0, 7·5, 8·0, and 9·0 for 4 days and urine was collected. Other specimens were also exposed to the experimental pH for 24 h and blood was sampled. Urine flow rate, urine and plasma pH showed a significant trend to increase with the increase of water pH. Urinary Na⁺ excretion rate also increased and ammonia urinary excretion rate decreased with the increase of water pH. There was a significant trend to decrease volume, ammonia, Cl⁻ and Na⁺ urinary excretion rate with increasing mass in fish exposed to all pH levels studied. Plasma ammonia levels showed a slight decrease in fish exposed to water pH from 4·0 to 8·0, but those exposed to water pH 9·0 presented the highest ammonia levels. Most plasma ions and urinary excretion changes observed in silver catfish exposed to acidic or alkaline water were similar to those already detected in rainbow trout Oncorhynchus mykiss . In addition, the kidney and urinary bladder might participate on acid–base balance in silver catfish, since urine pH changed according to plasma pH.     

The effects of capture, handling, confinement and ectoparasite load on plasma levels of cortisol, glucose and lactate in the coral reef fish Hemigymnus melapterus

Tropical labrids Hemigymnus melapterus sampled underwater had low plasma levels of cortisol, glucose, and lactate. Plasma cortisol levels were elevated by capture stress within 5–6 min, while glucose and lactate levels were not. Plasmalevels of cortisol and glucose increased after 2–4 h of handling and transport to the laboratory. Levels of cortisol and glucose fell with laboratory acclimation back to values similar to those found in wild fish. Parasitism by gnathiid isopods across an order of magnitude of isopod numbers had no effect on plasma levels of cortisol or glucose. Thus, H. melapterus has a stress response similar to that shown by temperate species, and relatively high parasite loads are not apparently stressful to fish in the wild. This may be related to the counterproductive effects of physiological stress responses on the immune system or behaviour-modulated processes that counter parasitic invasion.     

Stress and survival of small pike-perch Sander lucioperca (L.) after trawling and chilling

The mortality and stress responses (plasma cortisol concentration) of undersized (total length, L _T, <370 mm) pike-perch Sander lucioperca caught as by-catch in a trawl fishery were assessed. The effects of three different holding methods on pike-perch caught by trawl were compared: (1) fish that were transferred to ambient-temperature water (15·0–21·4° C) immediately after capture and not exposed to chilling water, (2) fish held in chilling water for 10 min and (3) fish exposed to chilling water for 2 h. The sample fish were held and monitored in individual plastic restrainers at ambient-water temperature for 48 h after each of the handling procedures. Total mortality was similar for fish held in chilling tanks for 10 min (27·2% total mortality) compared to fish that were only held in ambient-temperature water (28·2% mortality). Total mortality reached 91·3% when fish were held in chilled water for 2 h. Mortality and plasma cortisol concentrations correlated inversely with size (96–368 mm L _T) and directly with lake-water temperature (15·0–21·4° C) after treatment. In addition, the effect of chilling was observed in elevated cortisol concentration and delayed start of recovery. The results show that quickly removing (<10 min) undersized pike-perch from chilled water could markedly improve the survival of released fish. Due to delayed recovery, however, fish should be allowed to recover (30–40 min) in a deck tank before release.     

Stress caused by the 'mark-recapture' method to Coregonus albula (L)

The aim of this study was to establish if capture and marking caused stress to C. albula , and if a 'mark-recapture' method was suitable for the estimation of a population of C. albula . The blood lactate and glucose levels and the liver and muscle glycogen content were determined during the initial handling and subsequent recovery of the fish in the autumn and spring.
The blood lactate content did not increase to the critical level. Usually the values were distinctly less than 100 mg %, but the blood glucose content increased throughout the whole experiment (24 h) until it reached 250 mg %. The glycogen content in liver and muscle decreased. The liver glycogen of female C. albula was below 0·3 % in autumn, while the values for males were 3–5 %. In spring, the corresponding values were about 0·5 % in both sexes. During handling, the critical period was when the glycogen reserves in the liver became negligible, and female C. albula in autumn had the greatest risk of dying because of their low liver glycogen content.
Spring seemed to be a better season for marking than the autumn. A steady increase in blood glucose indicated there was also stress in spring and the marking time in both seasons should be less than two hours.     

Haematological effects of stress on a teleost, Esox lucius L.

Stress produces a haemoconcentration, elevated blood lactate, increased glucose concentrations and alters the plasma electrolyte balance in two groups (brackish- and freshwater) of the northern pike, Esox leucius L., after one month's starvation. A method for dorsal aorta catheterization and a receptacle for cannulating fish is described.
The blood glucose level of the freshwater pike was twice that of the brackish-water group, and the plasma sodium and magnesium concentrations in the brackish-water pike were significantly higher. The haematocrit, haemoglobin and blood lactic acid concentrations were higher in freshwater pike. The plasma potassium and calcium concentrations in the two groups did not differ.
Haemoconcentration due to stress by handling for 1.5 min was shown by changes in haematocrit and haemoglobin values. The haemoglobin concentration returned to normal in freshwater pike after 4 h but in brackish-water pike after 12 h.
As a result of handling, the blood lactic acid level rose steeply and required 12 h to return to normal.
The blood glucose concentration rose to its maximum value within 1 h of handling and required two days to return to normal.
The plasma sodium level remained stable after handling, but the potassium level was erratic. In brackish-water, the potassium concentration of the pike remained high for 12 h after stress, but in the freshwater group, after a rise, the concentration fell to below the initial level within 4 h. The changes of the potassium concentrations in relation to sampling time are discussed. The changes in the divalent ion concentrations were marked and similar in the two groups; with an increase lasting 1–4 h and then a fall below the initial level, which was regained after two days.     

Fasting modifies Aroclor 1254 impact on plasma cortisol,glucose and lactate responses to a handling disturbance in Arctic charr

Integrated effects of polychlorinated biphenyl (PCB) and nutritional status on responses to handling disturbance were investigated in the Arctic charr (Salvelinus alpinus). The fish were orally contaminated with Aroclor 1254 and held either with or without food for 5 months before they were subjected to a 10-min handling disturbance. Food-deprived fish were given 0, 1, 10 or 100 mg PCB kg(-1) and the fed fish 0 or 100 mg PCB kg(-1). Plasma cortisol, glucose and lactate levels were measured at 0 (pre-handling), 1, 3, 6 and 23 h after the handling disturbance. Food-deprived control fish had elevated plasma cortisol levels compared with fed fish before handling. These basal cortisol levels were suppressed by PCB in food-deprived fish, and elevated by PCB in fed fish. The immediate cortisol and glucose responses to handling disturbance were suppressed by PCB in a dose-dependent way in food-deprived fish. Although these responses were also lowered by PCB in the fed fish, the effect was much less pronounced than in food-deprived fish. There were only minor effects on plasma lactate responses. Our findings suggest that the stress responses of the Arctic charr are compromised by PCB and that the long-term fasting, typical of high-latitude fish, makes these species particularly sensitive to organochlorines such as PCB.     

Cortisol and glucose responses in juvenile brown trout subjected to a fluctuating flow regime in an artificial stream

In an artificial stream channel, wild 1 year old brown trout Salmo trutta were exposed to fluctuations in flow and water level to simulate hydro-peaking conditions downstream of a hydropower installation. Blood plasma cortisol concentrations reached a maximum of 59·4 ± 35·3 ng ml⁻¹ (mean ± 95% CL) 2h after the end of down-ramping. Return to the pre-exposure cortisol level was achieved within 6 h. When subjected to daily cyclical fluctuations over 7 days, plasma cortisol levels were significantly elevated (61·3 ± 26·8 ng ml⁻¹) on the first day compared to undisturbed fish (4·9 ± 3·7 ng ml⁻¹). On the fourth and seventh day, no elevation in plasma cortisol above control levels was observed. No changes in blood glucose that could be attributed to the stressor were found. There was no correlation between plasma cortisol and blood glucose levels. The short-lived cortisol response to daily fluctuations indicates a rapid habituation to this Stressor.     

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.     

Endocrine and metabolic responses to stress in a laboratory population of the tropical damselfish Acanthochromis polyacanthus

The effects of confinement and exercise on the stress response of the spiny damselfish Acanthochromis polyacanthus were investigated in a laboratory stock of fish. Cultured spiny damselfish had basal plasma cortisol values (<16 ng ml⁻¹) similar to those found in wild fish, and basal plasma glucose and lactate levels that were similar to those found in other teleosts. Plasma cortisol concentrations increased in response to stress with a latency period of 5–10 min. Removal of the stressor resulted in partial recovery of cortisol levels by 24 h. Plasma glucose levels increased in response to stress in all experiments with significant increases occurring within 15 min of the imposition of stress. Elevations in plasma glucose concentrations were not initially reflected in changes in liver or muscle glycogen content, with significant reductions in liver glycogen concentrations only occurring in response to extended periods of stress. In contrast to many temperate species, plasma lactate concentrations did not consistently increase in response to stress, suggesting that the stress response in spiny damselfish is not strongly characterized by anaerobiosis.     

Effects of dietary amino acids and repeated handling on stress response and brain monoaminergic neurotransmitters in Senegalese sole (Solea senegalensis) juveniles

The present study aimed to assess the effects of increased availability of dietary amino acids (AA) on brain monoamine neurotransmitters and the metabolic processes resulting from stressful situations in fish. Senegalese sole (Solea senegalensis) juveniles (24.2 ± 0.4 g wet mass) were weekly subjected to an acute handling stressor (HDLG) or remained undisturbed (CTL). Additionally, both treatments were fed a control or a high protein (HP) diet (CTL, CTL HP, HDLG and HDLG HP). The HP diet slightly increased the levels of digestible indispensable AA, together with tyrosine and cysteine. Repeated handling induced a stress response after 14 and 28 days in fish held at both HDLG and HDLG HP treatments. While dietary treatment and handling stress activated the serotonergic system at 14 days, these effects were not observed after 28 days. In addition, the HP diet minimized the decrease in plasma indispensable AA due to repeated handling stress after 28 days. It was concluded that HP diet decreased post-stress plasma glucose and lactate levels in HDLG HP specimens only at 14 days of treatment. Moreover, dietary treatment was also effective in stimulating DA synthesis and release, thus dietary phenylalanine supplementation can increase DA biosynthesis in fish.     

Body temperature and seawater adaptation in farmed Atlantic salmon and rainbow trout during prolonged chilling

The core temperature of the rainbow trout Oncorhynchus mykiss (3·5 kg) dropped to 1·0° C during the first 6 h of chilling at 0·5° C, remained stable until 24 h, and dropped significantly to 0·7° C after 39 h. Blood plasma osmolality increased and muscle moisture content decreased gradually with increasing chilling time. After 39 h of chilling, the rainbow trout experienced 40 mosmol l^-1 higher blood plasma osmolality and 2·8% less muscle moisture content compared with initial values. In the Atlantic salmon Salmo salar (5·3 kg), core temperature dropped to 1·3° C and blood plasma osmolality increased significantly during the first 6 h of chilling at 0·5° C, but remained relatively stable throughout the rest of the experimental period. After 39 h of chilling, the salmon experienced 20 mosmol l^-1 higher blood plasma osmolality and 0·5% less muscle moisture content compared with initial values. In rainbow trout muscle moisture content was inversely related to blood plasma osmolality indicating reduced seawater adaptation with increasing hours of chilling. No such relationship was observed in the Atlantic salmon. Hence, changes in plasma osmolality and muscle moisture in the Atlantic salmon do not indicate osmoregulatory failure since the new levels, once established, were maintained throughout the chilling time.     

Plasma cortisol and 17β-oestradiol levels in roach exposed to acute and chronic stress

Plasma cortisol levels were measured as an indicator of physiological stress in roach subjected to brief handling, or to a 14-day period of confinement, and in undisturbed control fish, during winter (water temperature 5° C) and summer (16° C), at which time plasma 17 β-oestradiol levels were also determined. Cortisol levels in undisturbed roach were low (mean 8·1 ng ml⁻¹ at 5° C; 1·4 ng ml⁻¹ at 16° C) and both handling and handling+confinement elevated blood cortisol levels significantly to 400 and 140 ng ml⁻¹, respectively (at 5° C) and 700 and 600 ng ml⁻¹, respectively (at 16) C). Blood cortisol levels had almost returned to baseline within 4 h following handling alone but in fish subjected to handling and prolonged confinement cortisol levels remained elevated for up to 168 h. Differences in baseline and poststress levels of cortisol, and in the rate of recovery from acute stress, were observed at the two different temperatures and the possible factors underlying these differences are discussed. Circulating levels of 17 β-oestradiol were reduced significantly within 24 h of exposure to either acute handling or chronic confinement indicating that the reproductive endocrine system in roach is sensitive to disruption by stressors.     

Influence of rearing temperature during the larval and nursery periods on growth and sex differentiation in two Mediterranean strains of Dicentrarchus labrax

European sea bass Dicentrarchus labrax of the north‐western (NW) and south‐eastern (SE) Mediterranean Sea strains were exposed to different temperatures (13, 17 or 21° C) during the larval rearing (11–51 days post hatching, dph) or nursery periods (55–95 dph), in order to examine the effects of temperature on sex differentiation and subsequent growth during the first year of life. Higher growth was observed during exposure to higher temperatures, but fish of the NW strain exposed to 13 or 17° C during larval rearing exhibited compensatory growth once exposure to the lower temperatures finished, and as a result their final size at 300 dph was similar or greater to the group exposed to 21° C. Fish exposed to 17° C during the nursery period also had similar size to fish exposed to 21° C after 300 days of rearing, but the fish exposed to 13° C remained significantly smaller (ANOVA, n  = 55–100, P  < 0·05). There were significant differences in the sex ratio among the fish exposed to different temperatures during the two periods of rearing, with high temperature (21° C) resulting in a significantly higher percentage of males in the population, both in the NW (ANOVA, n  = 2, P  < 0·04) and SE populations (ANOVA, n  = 2, P  < 0·01). The masculinization effect of high temperature was significantly stronger during the larval rearing stage, both in the NW (ANOVA, n  = 2, P  < 0·005) and SE populations (ANOVA, n  = 2, P  < 0·01). None of the temperature manipulations could produce 100% females, suggesting that there is a part of the genetic component in sex differentiation which is not labile to environmental influence.     

Estimating sampling bias when using electrofishing to catch stone loach

Point abundance sampling, using electrofishing, for stone loach Barbatula barbatula in two habitats, riffle and pool, showed that there was no difference in density between the two habitats. The density of fish increased significantly (1·8 fold; P  < 0·001) when the substratum of small cobbles (128–256 mm) was removed and electrofishing repeated.     

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.