Plasma catecholamines and erythrocyte swelling following capture stress in a marine teleost fish

Plasma concentrations of adrenaline and noradrenaline were measured at rest from cannulated fish and following net capture. Adrenaline and noradrenaline concentrations in capture-stressed fish averaged 36,740 pmol l-1 and 38,860 pmol l-1 respectively, whereas resting values were less than 200 pmol l-1 for both amines. Erythrocyte swelling and raised blood lactate were evident in stressed fish. In vitro effects of 5 mmol l-1 adrenaline on erythrocyte suspensions suggested that the catecholamine had a direct effect on erythrocyte volume. The significance of these results is discussed in relation to the oxygen transport properties of the blood.     

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.     

Immediate and delayed effects of gill-net capture on acid-base balance and intramuscular lactate concentration of gummy sharks, Mustelus antarcticus

Many sharks are captured as untargeted by-catch during commercial fishing operations and are subsequently discarded. A reliable assessment of the proportion of discarded sharks that die post-release as a result of excessive physiological stress is important for fisheries management and conservation purposes, but a reliable physiological predictor of post-release mortality has not been identified. To investigate effects of gill-net capture on the acid-base balance of sharks, we exposed gummy sharks, Mustelus antarcticus, to 60 min of gill-net capture in a controlled setting, and obtained multiple blood and muscle tissue samples during a 72-h recovery period following the capture event. Overall mortality of gummy sharks was low (9%). Blood pH was significantly depressed immediately after the capture event due to a combination of respiratory and metabolic acidosis. Maximum concentrations of plasma lactate (9.9 ± 1.5 mmol L(-1)) were measured 3h after the capture event. Maximum intramuscular lactate concentrations (37.0 ± 4.6 μmol g(-1)) were measured immediately after the capture event, and intramuscular lactate concentrations were substantially higher than plasma lactate concentrations at all times. Sharks in poor condition had low blood pH and high intramuscular lactate concentration, but blood pH does not appear to be a reliable predictor of survival. Suitability of intramuscular lactate concentration as predictor of delayed mortality deserves further investigation.     

Time course of osmoregulatory, metabolic, and endocrine stress responses of Pacific halibut following a 30-min air exposure

Endocrine, metabolic and osmoregulatory changes in Pacific halibut upon capture and at intervals following a 30‐min air exposure were measured. Concentrations of cortisol, sodium, and chloride in plasma and serum peaked two hours after the stressor. Thirty minutes after a 30‐min air exposure concentrations of glucose in plasma had increased significantly from levels obtained immediately after the stressor and remained elevated for up to 4 h. Plasma lactate was also elevated 30 min after the stress treatment and lactate concentrations increased significantly at each subsequent sampling interval, 2 and 4 h. There was a significant linear increase in plasma lactate for fish sampled from 1.5 to 6.5 h after capture. Incidence of delayed mortality was low; of 22 experimentally stressed animals only one died over a 10‐day monitoring period. These data illustrate the difficulties in using single time‐point plasma indices of stress to assess condition of animals after capture since the time‐course of physiological changes associated with the stress response vary with parameter measured and may take hours to be fully expressed.     

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

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

Acid inactivation of short-lived rat liver enzymes.

The stabilities of nine rat liver cytosol enzymes were compared at a variety of pH values. The cytosol enzymes studied were (a) those with half-lives in vivo of 3 days or longer: lactate dehydrogenase, arginase, glyceraldehyde phosphate dehydrogenase and alanine aminotransferase, (b) those with half-lives in vivo shorter than 2 days; glucokinase, dihydroorotase, serine dehydratase and tyrosine aminotransferase and (c) catalase, which has an intermediate half-life of 2.5 days for the protein protion. All the enzymes were stable in vitro at neurtal and alkaline pH values. However, at acidic pH values (pH 4): the long-lived enzymes (a) were stable; the short-lived enzymes (b) were completely inactivated with one exception; and catalase was partially inactivated. Tyrosine aminotransferase was the exception in that it is a short-lived enzyme in vivo but stable under all conditions tested in vitro. The finding that long-lived enzymes are stable in an acid milieu and short-lived enzymes are generally unstable was only observed if certain ligands (NAD+, pyridoxal 5'-phosphate, Mn2+, amino acids) were added to the invitro system. Lysosomal extracts did not accelerate the rate of inactivation of any cytosol enzyme in acidic solutions. These results indicate that if degradation of intracellular enzymes occurs in lysosomes, acid inactivation and denaturation of enzymes may be the initial event in determining the functional half-lives of the enzymes in vivo.     

Effects of acepromazine on capture stress in roe deer (Capreolus capreolus)

The aim of this study was to evaluate effect of a short-acting neuroleptic (acepromazine) on capture stress response in roe deer (Capreolus capreolus). Sixteen roe deer were captured by drive-nets in the winters of 1998, 1999, and 2001. Roe deer were divided into two groups: animals in the treatment group received an intramuscular injection of acepromazine (0.093 mg/kg +/- 0.003 SEM; n = 8) while animals in the control group (n = 8) did not receive tranquilizer. Heart rate and body temperature, as well as hematologic and biochemical indicators of stress, were used to evaluate effect of the neuroleptic over 3 hr. Heart rate decreased over time after capture in both groups (P < 0.05), but stabilized sooner in the treated roe deer (75 min after capture) than in the controls (105 min after capture). Body temperature decreased over 45 min and then stabilized in both groups (P < 0.05). Comparisons of blood parameters revealed significantly lower red blood cell count (RBC), lymphocyte count, hemoglobin concentration, packed cell volume (PCV), and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK), and lactate dehydrogenase (LDH) activities in tranquilized animals compared with controls (at least P < 0.05). A reduction in PCV, lymphocyte count, and serum cortisol concentrations (at least P < 0.05) and an increase in serum creatinine levels (P < 0.05) were recorded over time in control animals, while a reduction in RBC and hemoglobin concentration (at least P < 0.05) and an increase in serum urea concentrations (P < 0.05) over time were observed in the treated group. Finally, a decrease in serum lactate and potassium levels and an increase in CK, AST, ALT, and LDH activities were recorded over time in both groups. Results obtained showed the suitability of using acepromazine in capture operations in order to reduce stress response and prevent its adverse effects in roe deer. The beneficial effect was not only due to the sedative effect of acepromazine, but also to peripheral vasodilatation.     

Effects of handling stress on plasma enzymes in harp seals, Phoca groenlandica.

Three harp seal pups, Phoca groenlandica, were captured on the ice of the Gulf of St. Lawrence, and subjected to 3 h of transportation and handling stress. The activities of creatine kinase (CK), aspartate aminotransferase (AspAT), aldolase, alanine aminotransferase, gamma glutamyl transpeptidase, and leucine aminopeptidase were determined in serial blood samples collected for 4 d following the stress episode. Marked elevation of plasma CK activity was observed 3 h after capture. Values returned to normal in 12 h in two seals, and by 24 h in the third. Slight elevations in AspAT were also noted; the remaining enzymes were unaffected. Plasma CK is recommended as a sensitive indicator of handling stress in seals.     

Plasma volume determination by use of enzyme dilution in the dog

The use of enzymes for plasma volume determination in the dog was investigated. Plasma volume was determined from dilution of intravenously injected enzymes and compared with results obtained by high molecular weight fluorescein-labelled dextran. Aspartate aminotransferase and alanine aminotransferase from liver tissue gave good results, but the activities of creatine kinase and lactate dehydrogenase from heart tissue were inhibited in plasma, resulting in an overestimation of plasma volume. Enzymes offer a useful alternative to methods presently used since no radioactive isotopes are needed. In laboratory animals the enzyme preparations are readily obtainable and there is minimal risk of immunological complications arising after repeated determinations.     

Effects of capture and recovery on plasma levels of cortisol, lactate and gonadal steroids in a natural population of rainbow trout

Rainbow trout were captured by angling from a run of spawning fish on the Tongariro River in northern New Zealand, to examine the effects of catch and release angling on stress and reproductive parameters. Fish were blood sampled immediately after capture at playing times of <5 or 15 min, or after 1 or 24 h of recovery in stream enclosures. Plasma samples were assayed for cortisol (F), lactate, testosterone (T), 17β-oestradiol (E₂), and 17a,20β-dihydroxy-4-pregnen-3-one (17,20βP). Plasma F levels were similar to those of hatchery stocks of rainbow trout, at capture, and became significantly elevated 1 h after capture. Plasma F was still clevated in some fish 24 h after capture. Plasma lactate levels began to increase 15 min after capture, were further elevated 1 h after capture, and had returned to normal 24 h after capture. We proposed that metabolic recovery had occurred but that some animals were still experiencing some degree of stress, possibly in response to holding conditions in the river. Both plasma T and E₂ were depressed 24 h after capture, whereas there was no change in plasma 17,20βP. This is consistent with other findings showing that acute stress is associated with depression of plasma levels of T and E₂. There was no mortality as a result of capture or any of the handling protocols. We conclude that catch and release angling will result in negligible mortality, but may have an inhibitory effect on some reproductive processes.     

Effect of polychlorinated biphenyls (Delor 103) on haematological and enzyme parameters of the rainbow trout Oncorhynchus mykiss

Rainbow trout at a weight of 223+/-12 g (mean+/-SD) were experimentally injected with a technical mixture of Delor 103 to evaluate the red blood cell indices (red blood cell count, haematocrit, haemoglobin, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration) and some biochemical and enzyme parameters of the blood plasma (total protein, glucose, inorganic phosphate, total calcium, sodium, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase). Delor 103, administered by the i.p. route at a concentration of 0.24 g kg(-1) 120 h(-1), caused an increase in the red blood cell counts, haematocrit values, haemoglobin concentrations, inorganic phosphate, alanine aminotransferase and lactate dehydrogenase. The sodium level fell. The fish injected with Delor 103 showed a relative decrease in the lymphocyte count and a relative increase in the count of neutrophile band forms.     

Influence of water temperature and net tending frequency on the condition of fish bycatch in a small-scale inland commercial fyke net fishery

To date, most studies of commercial fisheries bycatch have focused on mortality at time of capture as an endpoint. However, sub-lethal indicators of organismal condition have the potential to reveal mechanisms associated with mortality (both at time of capture and post-release) and opportunities for improving fish welfare. In this study, we simulated commercial fishing efforts in inland lakes with fyke nets during a typical fishing season (early April to late June) in southeastern Ontario, Canada, where bycatch of non-target fish species had previously been documented. Using non-target gamefish (i.e., largemouth bass [Micropterus salmoides, Lacépède], northern pike [Esox Lucius, L.]), as well as a target species (i.e., bluegill [Lepomis macrochirus, Rafinesque]), we examined the sub-lethal consequences of capture (e.g., blood physiology, reflex impairment, and injury) and compared the effects of being retained in the net for two different durations (i.e., two or six days) over a range of water temperatures (i.e., 3–28 °C). Sub-lethal physiological disturbances (i.e., blood glucose and lactate) in largemouth bass and bluegill tended to be greater at higher water temperatures. However, fish retained for six days generally did not exhibit greater stress than those retained for two days, with the exception of plasma glucose in largemouth bass. Reflex impairment was similar among temperature and retention periods. Fish retained in nets experienced a range of injuries (including fin frays, scale loss, and mouth damage) that had the potential to facilitate the development of opportunistic pathogenic infections. Greater incidences of injury on fish bycatch tended to be associated with higher temperatures and longer retention. To reduce physiological disturbances and injury that could lead to delayed mortality, we suggest that regulations for inland commercial fishers require them to check their nets more frequently as water temperatures increase. We suggest that future studies of bycatch incorporate sub-lethal endpoints given that they serve as an objective measure of fish welfare and can provide quantitative mechanistic information to support management actions.     

Stress induced by hooking, net towing, elevated sea water temperature and air in sablefish: lack of concordance between mortality and physiological measures of stress

In a series of laboratory studies designed to simulate bycatch processes, sablefish Anoplopoma fimbria were either hooked for up to 24 h or towed in a net for 4 h and then subjected to an abrupt transfer to elevated sea water temperature and air. Mortality did not result from hooking or net towing followed by exposure to air, but increased for both capture methods as fish were exposed to elevated temperatures, reflecting the magnifying effect of elevated temperature on mortality. Hooking and exposure to air resulted in increased plasma cortisol and lactate concentrations, while the combination of hooking and exposure to elevated temperature and air resulted in increased lactate and potassium oncentrations. In fish that were towed in a net and exposed to air, cortisol, lactate, potassium and sodium concentrations increased, but when subjected to elevated temperature and air, no further increases occurred above the concentrations induced by net towing and air, suggesting a possible maximum of the physiological stress response. The results suggest that caution should be exercised when using physiological measures to quantify stress induced by capture and exposure to elevated temperature and air, that ultimately result in mortality, since the connections between physiological stress and mortality in bycatch processes remain to be fully understood.     

Experimental determination of net protein charge and A(tot) and K(a) of nonvolatile buffers in human plasma.

The mechanism for an acid-base disturbance can be determined by using the strong ion approach, which requires species-specific values for the total concentration of plasma nonvolatile buffers (Atot) and the effective dissociation constant for plasma weak acids (Ka). The aim of this study was to experimentally determine Atot and Ka values for human plasma by using in vitro CO2 tonometry. Plasma Pco2 was systematically varied from 25 to 145 Torr at 37 degrees C, thereby altering plasma pH over the physiological range of 6.90-7.55, and plasma pH, Pco2, and concentrations of quantitatively important strong ions (Na+, K+, Ca2+, Mg2+, Cl-, lactate) and buffer ions (total protein, albumin, phosphate) were measured. Strong ion difference was estimated, and nonlinear regression was used to calculate Atot and Ka from the measured pH and Pco2 and estimated strong ion difference; the Atot and Ka values were then validated by using a published data set (Figge J, Rossing TH, and Fencl V, J Lab Clin Med 117: 453-467, 1991). The values (mean +/- SD) were as follows: Atot = 17.2 +/- 3.5 mmol/l (equivalent to 0.224 mmol/g of protein or 0.378 mmol/g of albumin); Ka = 0.80 +/- 0.60 x 10-7; negative log of Ka = 7.10. Mean estimates were obtained for strong ion difference (37 meq/l) and net protein charge (13+.0 meq/l). The experimentally determined values for Atot, Ka, and net protein charge should facilitate the diagnosis and treatment of acid-base disturbances in critically ill humans.     

The blood chemical status of Atlantic cod Gadus morhua following capture by jig and demersal longline with differential hook removal methods

Common haematological [haematocrit (Hct)], primary (serum cortisol) and secondary (serum glucose and plasma lactate) analytes were utilized to compare blood biochemical status of Gadus morhua captured rapidly by jig with that of G. morhua captured by commercial demersal longline. In general, the physiological status of G. morhua, despite blind hook times, was significantly more disrupted (pronounced haemo-concentration and significantly elevated concentrations of cortisol, glucose and lactate) following longline capture relative to capture by jig, while no differences were detected among longline-caught fish as a function of dehooking method (or concomitant extent of overt physical trauma). Blood profiles from the more stressed G. morhua, a possible function of more extended longline hook times, were similar to the most stressed values reported for this species. The results also demonstrate that, although acute blood biochemical status is an effective gauge of relative stress, it does not reflect physical injury status, which has been shown to exert a strong influence on delayed mortality in previous studies in this species. Thus, acute blood chemical status alone may not be the most complete predictor of mortality. Future studies should evaluate physiological repercussions from capture-handling against physical trauma during more extended post-release periods for this species.     

Enzyme activities in plasma, kidney, liver, and muscle of five avian species

Activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine phosphokinase (CPK), and lactate dehydrogenase (LDH) were determined in plasma, kidney, liver, and muscle from five species of captive birds. Few differences occurred in plasma activities between sexes but considerable differences occurred between species. All five enzymes were detected in each of the tissues sampled. Relative enzyme activities in liver, kidney, and muscle were similar for each species. CPK activity was much higher in muscle than in liver or kidney and, of the five enzymes studied, may be the best indicator of muscle damage. Most of the other enzymes were more evenly distributed among the three tissues, and no organ-specific enzyme could be identified for liver or kidney. Because of interspecific variations in plasma enzyme activities, it is important to establish baseline values for each species to ensure accurate interpretation of results.     

Sodium selenite modulates tumour marker indices in N-nitrosodiethylamine-initiated and phenobarbital-promoted rat liver carcinogenesis

The effect of sodium selenite (Se) was investigated against two-stage rat liver carcinogenesis initiated by a single intraperitoneal injection of N-nitrosodiethylamine (DEN, 200 mg kg(-1) i.p.) followed by promotion with phenobarbital (PB, 0.05%) in a basal diet. Se (4 p.p.m.) was administered per os daily throughout the entire experiment, before the initiation, or during the promotion stage. The plasma, liver (hepatoma and surrounding tissue) and kidney tissue were investigated biochemically for lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and 5'-nucleotidase. These enzyme activities were increased (p < 0.001) in plasma of hepatoma-bearing rats compared with normal control rats. The elevation of these enzyme activities in plasma was indicative of the persistent deteriorating effect of DEN in cancer-bearing animals. Aminotransferase levels were decreased in hepatoma and surrounding liver tissue, whereas lactate dehydrogenase, alkaline phosphatase and 5'-nucleotidase were increased in the cancer condition. These enzyme activities were reversed to near normal control values in animals treated with Se. It is apparent that the beneficial effect of Se is primarily exerted on the initiation phase and secondarily during the promotion stage of DEN-initiated rat liver carcinogenesis. The analysis of marker enzyme activities taken together with our previous findings clearly indicates the antitumour efficacy of sodium selenite on DEN-induced hepatoma animals.     

Changes in haematological parameters associated with capture and captivity of the marine teleost, Pleuronectes platessa L

After capture by trawling, the blood parameters of plaice (Pleuronectes platessa L.) are perturbed for up to 5 days post-capture. Whole blood values recovered from an initial stress-induced haemoconcentration within 12 hr. There is a marked hyperglycaemia following capture: blood glucose concentration increased four-fold to 87.92 +/- 10.41 mg/100 ml (N = 6) after 12 hr and remained elevated for 3-4 days before returning to normal values. Monovalent blood electrolytes (Na+, K+, Cl-) significantly increased during the initial stages post-capture (4-10 hr) but then recovered. The divalent cations (Ca2+, Mg2+) similarly increased but for a longer period (24-72 hr). Liver and muscle glycogen concentrations were very variable during the recovery period. All blood parameters achieved stable values within 5 days of capture. This study provides comprehensive haematological data on post-trawl recovery and tank-acclimation in plaice, for up to 28 days following capture.     

Effects of acute hypobaric hypoxia on acid-base status of fowl blood

Arterial blood Po/, Pco2, lactate levels and Cl- ion concentration as well as pH were measured on the time course in chickens (Gallus domesticus) as they settled in normoxic conditions and during exposure to acute hypobaric hypoxia (Pb = 450 Torr). Hypoxia provoked at first a CO2 increased output from blood and a brief stage of deep metabolic acidosis during which lactate levels suddenly increased. This acidosis was then compensated producing a return to the initial pH and a decrease in [HCO3-] + [CO3(2-)] after 60 min. Subsequently respiratory alkalosis associated with an increase in [HCO-3] + [CO3(2-)], a decrease in cl- ion concentration and a small decrease in lactate levels were observed. Prolonged exposure to hypoxia (16 h) resulted in a new return to the initial pH, a decrease in concentration of [HCO3-] + [CO3(2-)] and a high lactate level. The hematocrit value, the Hb concentration, and the plasma Na+, K+, Ca++ and Mg++ ion concentration did not change significantly.     

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.