Effects of dispersant-treated oil upon behavioural and metabolic parameters of the anti-predator response in juvenile European sea bass (Dicentrarchus labrax).

Acute exposure to oil and oil dispersants can cause a wide range of physiological dysfunctions in marine fish species and evidences for consequences on behaviour are also increasing. In response to the presence of predators or to food availability, the modulation of locomotor activity and schools' behaviour enable fish to maximize their survival rates. However, the degree to which this regulatory process is affected by exposure to oil and/or dispersants is yet unknown. Here we investigated the effect of a 62-h experimental exposure to dispersant-treated oil on the behavioural (shoal cohesion, spontaneous activity) and metabolic (oxygen consumption) responses to simulated predation in juvenile European sea bass, Dicentrarchus labrax L. Our results suggest that exposure to petroleum hydrocarbons may affect negatively individual fitness through impaired ability to respond to predation. Shoal cohesion was not affected, but fish swimming activity was higher than control individuals under predation pressure and the amplitude of their metabolic response was significantly reduced. Fish recovered from alteration of their metabolic response 7 days post-exposure. Additionally, a strong habituation component was observed in C fish and the absence of such pattern in E fish suggest altered capacity to habituate over time to the surrounding environment and possible impairments of the related cognitive performances. Altogether, our data show that juvenile sea bass exposed to oil exhibit transient physiological dysfunctions and impairments of complex behaviours that may have major population-level consequences.

A three-phase excess post-exercise oxygen consumption in Atlantic salmon Salmo salar and its response to exercise training.

The recovery of oxygen uptake to the standard metabolic rate (SMR) following exhaustive chasing exercise in Atlantic salmon Salmo salar parr occurred in three phases (rapid, plateau and slow). The initial recovery phase lasted 0·7 h and contributed 16% to the total excess post-exercise oxygen consumption (EPOC). It was followed by a longer plateau phase that contributed 53% to the total EPOC. The slow recovery phase that completed recovery of SMR, which has not been reported previously, made a 31% contribution to the total EPOC. The plasticity of EPOC was demonstrated in exercise-trained fish. Exercise training increased EPOC by 39% when compared with control fish (mean ± S.E., 877·7 ± 73·1 v. 629·2 ± 53·4 mg O2 kg-1 , d.f. = 9, P < 0·05), with the duration of the plateau phase increasing by 38% (4·7 ± 0·58 v. 3·4 ± 0·16 h, d.f. = 9, P < 0·05) and the contribution of the slow phase to the total EPOC increasing by 80% (173·9 ± 23·9 v. 312·5 ± 50·4 mg O2 kg-1 , d.f. = 9, P < 0·05). As a result, the combination of the plateau and slow phases of exercise-trained fish increased by 47% compared with control fish (756·6 ± 71·4 v. 513·6 ± 43·1 mg O2 kg-1 ; d.f. = 9, P = 0·01). To substantiate the hypothesis that the plateau and slow recovery phase of EPOC was related to general metabolic recovery following exhaustive exercise, the time-course for recovery of SMR was compared with previously published metabolite recovery profiles. The final phase of metabolic recovery was temporally associated with the final phases of gluconeogenesis, lactate oxidation and muscle intracellular pH regulation. Therefore, the plasticity of the latter phase of EPOC agreed with the known effects of exercise training in fishes.

Divergence in physiological factors affecting swimming performance between anadromous and resident populations of brook charr Salvelinus fontinalis.

In this study, an anadromous strain (L) and a freshwater-resident (R) strain of brook charr Salvelinus fontinalis as well as their reciprocal hybrids, were reared in a common environment and submitted to swimming tests combined with salinity challenges. The critical swimming speeds (Ucrit ) of the different crosses were measured in both fresh (FW) and salt water (SW) and the variations in several physiological traits (osmotic, energetic and metabolic capacities) that are predicted to influence swimming performance were documented. Anadromous and resident fish reached the same Ucrit in both FW and SW, with Ucrit being 14% lower in SW compared with FW. The strains, however, seemed to use different underlying strategies: the anadromous strain relied on its streamlined body shape and higher osmoregulatory capacity, while the resident strain had greater citrate synthase (FW) and lactate dehydrogenase (FW, SW) capacity and either greater initial stores or more efficient use of liver (FW, SW) and muscle (FW) glycogen during exercise. Compared with R♀ L♂ hybrids, L♀ R♂ hybrids had a 20% lower swimming speed, which was associated with a 24% smaller cardio-somatic index and higher physiological costs. Thus swimming performance depends on cross direction (i.e. which parental line was used as dam or sire). The study thus suggests that divergent physiological factors between anadromous and resident S. fontinalis may result in similar swimming capacities that are adapted to their respective lifestyles.

Assessing chronic fish health: An application to a case of an acute exposure to chemically treated crude oil.

Human alteration of marine ecosystems is substantial and growing. Yet, no adequate methodology exists that provides reliable predictions of how environmental degradation will affect these ecosystems at a relevant level of biological organization. The primary objective of this study was to develop a methodology to evaluate a fish's capacity to face a well-established environmental challenge, an exposure to chemically dispersed oil, and characterize the long-term consequences. Therefore, we applied high-throughput, non-lethal challenge tests to assess hypoxia tolerance, temperature susceptibility and maximal swimming speed as proxies for a fish's functional integrity. These whole animal challenge tests were implemented before (1 month) and after (1 month) juvenile European sea bass (Dicentrarchus labrax) had been acutely exposed (48h) to a mixture containing 0.08gL(-1) of weathered Arabian light crude oil plus 4% dispersant (Corexit© EC9500A), a realistic exposure concentration during an oil spill. In addition, experimental populations were then transferred into semi-natural tidal mesocosm ponds and correlates of Darwinian fitness (growth and survival) were monitored over a period of 4 months. Our results revealed that fish acutely exposed to chemically dispersed oil remained impaired in terms of their hypoxia tolerance and swimming performance, but not in temperature susceptibility for 1 month post-exposure. Nevertheless, these functional impairments had no subsequent ecological consequences under mildly selective environmental conditions since growth and survival were not impacted during the mesocosm pond study. Furthermore, the earlier effects on fish performance were presumably temporary because re-testing the fish 10 months post-exposure revealed no significant residual effects on hypoxia tolerance, temperature susceptibility and maximal swimming speed. We propose that the functional proxies and correlates of Darwinian fitness used here provide a useful assessment tool for fish health in the marine environment.

Context dependency of trait repeatability and its relevance for management and conservation of fish populations.

Repeatability of behavioural and physiological traits is increasingly a focus for animal researchers, for which fish have become important models. Almost all of this work has been done in the context of evolutionary ecology, with few explicit attempts to apply repeatability and context dependency of trait variation toward understanding conservation-related issues. Here, we review work examining the degree to which repeatability of traits (such as boldness, swimming performance, metabolic rate and stress responsiveness) is context dependent. We review methods for quantifying repeatability (distinguishing between within-context and across-context repeatability) and confounding factors that may be especially problematic when attempting to measure repeatability in wild fish. Environmental factors such temperature, food availability, oxygen availability, hypercapnia, flow regime and pollutants all appear to alter trait repeatability in fishes. This suggests that anthropogenic environmental change could alter evolutionary trajectories by changing which individuals achieve the greatest fitness in a given set of conditions. Gaining a greater understanding of these effects will be crucial for our ability to forecast the effects of gradual environmental change, such as climate change and ocean acidification, the study of which is currently limited by our ability to examine trait changes over relatively short time scales. Also discussed are situations in which recent advances in technologies associated with electronic tags (biotelemetry and biologging) and respirometry will help to facilitate increased quantification of repeatability for physiological and integrative traits, which so far lag behind measures of repeatability of behavioural traits.

Responses by fishes to environmental hypoxia: integration through Fry's concept of aerobic metabolic scope.

The problem of understanding the effect of the environment on fish activities and performance, in any generalized way, remains intractable. Solving this issue is, however, a key to addressing contemporary environmental concerns. As suggested 20 years ago by W. H. Neill, the authors returned to the drawing board, using as a background the conceptual scheme initially proposed by F. E. J. Fry. They revisited the effect of ambient oxygen availability upon fish metabolism and clarified the definitions of limiting, critical and incipient lethal oxygen (ILO) levels. The concepts of oxy-conformer and oxy-regulator are revisited, and P. W. Hochachka's idea of scope for survival is explored. Finally, how the cardiovascular system contributes to the capacity of fishes to respond to the reduced oxygen availability is considered. Various hands-on recommendations and software (R scripts) are provided for researchers interested in investigating these concepts.

Physiological mechanisms underlying a trade-off between growth rate and tolerance of feed deprivation in the European sea bass (Dicentrarchus labrax).

The specific growth rate (SGR) of a cohort of 2000 tagged juvenile European sea bass was measured in a common tank, during two sequential cycles comprising three-weeks feed deprivation followed by three-weeks ad libitum re-feeding. After correction for initial size at age as fork length, there was a direct correlation between negative SGR (rate of mass loss) during feed deprivation and positive SGR (rate of compensatory growth) during re-feeding (Spearman rank correlation R=0.388, P=0.000002). Following a period of rearing under standard culture conditions, individuals representing 'high growth' phenotypes (GP) and 'high tolerance of feed deprivation' phenotypes (DP) were selected from either end of the SGR spectrum. Static and swimming respirometry could not demonstrate lower routine or standard metabolic rate in DP to account for greater tolerance of feed deprivation. Increased rates of compensatory growth in GP were not linked to greater maximum metabolic rate, aerobic metabolic scope or maximum cardiac performance than DP. When fed a standard ration, however, GP completed the specific dynamic action (SDA) response significantly faster than DP. Therefore, higher growth rate in GP was linked to greater capacity to process food. There was no difference in SDA coefficient, an indicator of energetic efficiency. The results indicate that individual variation in growth rate in sea bass reflects, in part, a trade-off against tolerance of food deprivation. The two phenotypes represented the opposing ends of a spectrum. The GP aims to exploit available resources and grow as rapidly as possible but at a cost of physiological and/or behavioural attributes, which lead to increased energy dissipation when food is not available. An opposing strategy, exemplified by DP, is less 'boom and bust', with a lower physiological capacity to exploit resources but which is less costly to sustain during periods of food deprivation.

Individual variation and repeatability in aerobic and anaerobic swimming performance of European sea bass, Dicentrarchus labrax.

Studies of inter-individual variation in fish swimming performance may provide insight into how selection has influenced diversity in phenotypic traits. We investigated individual variation and short-term repeatability of individual swimming performance by wild European sea bass in a constant acceleration test (CAT). Fish were challenged with four consecutive CATs with 5 min rest between trials. We measured maximum anaerobic speed at exhaustion (U(CAT)), gait transition speed from steady aerobic to unsteady anaerobic swimming (U(gt)), routine metabolic rate (RMR), post-CAT maximum metabolic rate (MMR), aerobic scope and recovery time from the CATs. Fish achieved significantly higher speeds during the first CAT (U(CAT)=170 cm s(-1)), and had much more inter-individual variation in performance (coefficient of variation, CV=18.43%) than in the subsequent three tests (U(CAT)=134 cm s(-1); CV=7.3%), which were very repeatable among individuals. The individual variation in U(CAT) in the first trial could be accounted for almost exclusively by variation in anaerobic burst-and-coast performance beyond U(gt). The U(gt) itself varied substantially between individuals (CV=11.4%), but was significantly repeatable across all four trials. Individual RMR and MMR varied considerably, but the rank order of post-CAT MMR was highly repeatable. Recovery rate from the four CATs was highly variable and correlated positively with the first U(CAT) (longer recovery for higher speeds) but negatively with RMR and aerobic scope (shorter recovery for higher RMR and aerobic scope). This large variation in individual performance coupled with the strong correlations between some of the studied variables may reflect divergent selection favouring alternative strategies for foraging and avoiding predation.

Acid-base regulatory ability of the cephalopod (Sepia officinalis) in response to environmental hypercapnia.

Acidification of ocean surface waters by anthropogenic carbon dioxide (CO(2)) emissions is a currently developing scenario that warrants a broadening of research foci in the study of acid-base physiology. Recent studies working with environmentally relevant CO(2) levels, indicate that some echinoderms and molluscs reduce metabolic rates, soft tissue growth and calcification during hypercapnic exposure. In contrast to all prior invertebrate species studied so far, growth trials with the cuttlefish Sepia officinalis found no indication of reduced growth or calcification performance during long-term exposure to 0.6 kPa CO(2). It is hypothesized that the differing sensitivities to elevated seawater pCO(2) could be explained by taxa specific differences in acid-base regulatory capacity. In this study, we examined the acid-base regulatory ability of S. officinalis in vivo, using a specially modified cannulation technique as well as (31)P NMR spectroscopy. During acute exposure to 0.6 kPa CO(2), S. officinalis rapidly increased its blood [HCO(3)(-)] to 10.4 mM through active ion-transport processes, and partially compensated the hypercapnia induced respiratory acidosis. A minor decrease in intracellular pH (pH(i)) and stable intracellular phosphagen levels indicated efficient pH(i) regulation. We conclude that S. officinalis is not only an efficient acid-base regulator, but is also able to do so without disturbing metabolic equilibria in characteristic tissues or compromising aerobic capacities. The cuttlefish did not exhibit acute intolerance to hypercapnia that has been hypothesized for more active cephalopod species (squid). Even though blood pH (pHe) remained 0.18 pH units below control values, arterial O(2) saturation was not compromised in S. officinalis because of the comparatively lower pH sensitivity of oxygen binding to its blood pigment. This raises questions concerning the potentially broad range of sensitivity to changes in acid-base status amongst invertebrates, as well as to the underlying mechanistic origins. Further studies are needed to better characterize the connection between acid-base status and animal fitness in various marine species.

Effects of feeding and hypoxia on cardiac performance and gastrointestinal blood flow during critical speed swimming in the sea bass Dicentrarchus labrax.

Previous studies have shown that if European sea bass are exercised after feeding, they can achieve a significantly higher maximum metabolic rate (MMR) than when fasted. They can meet combined metabolic demands of digestion (specific dynamic action, SDA) and maximal aerobic exercise, with no decline in swimming performance. If, however, exposed to mild hypoxia (50% saturation), bass no longer achieve higher MMR after feeding but they swim as well fed as fasted, due to an apparent ability to defer the SDA response. This study explored patterns of cardiac output (Q(A)) and blood flow to the gastrointestinal tract (Q(GI)) associated with the higher MMR after feeding, and with the ability to prioritise swimming in hypoxia. Sea bass (mean mass approximately 325 g, forklength approximately 27 cm) were instrumented with flow probes to measure Q(A) and Q(GI) during an incremental critical swimming speed (U(crit)) protocol in a tunnel respirometer, to compare each animal either fasted or 6h after a meal of fish fillet equal to 3% body mass. Feeding raised oxygen uptake (M(O2)) prior to exercise, an SDA response associated with increased Q(A) (+30%) and Q(GI) (+100%) compared to fasted values. As expected, when exercised the fed bass maintained the SDA load throughout the protocol and achieved 14% higher MMR than when fasted, and the same U(crit) (approximately 100 cm s(-1)). Both fed and fasted bass showed pronounced increases in Q(A) and decreases in Q(GI) during exercise and the higher MMR of fed bass was not associated with higher maximum Q(A) relative to when fasted, or to any differences in Q(GI) at maximum Q(A). In hypoxia prior to exercise, metabolic and cardiac responses to feeding were similar compared to normoxia. Hypoxia caused an almost 60% reduction to MMR and 30% reduction to U(crit), but neither of these traits differed between fed or fasted bass. Despite hypoxic limitations to MMR and U(crit), maximum Q(A) and patterns of Q(GI) during exercise in fasted and fed bass were similar to normoxia. Estimating GI oxygen supply from Q(GI) indicated that the ability of bass to prioritise aerobic exercise over SDA when metabolically limited by hypoxia was linked to an ability to defer elements of the SDA response occurring outside the GI tract.

Sublethal concentrations of ammonia impair performance of the teleost fast-start escape response.

The fast-start escape response in fish is essential for predator avoidance, but almost nothing is known about whether sublethal concentrations of pollutants can impair this reflex. Ammonia, a pervasive pollutant of aquatic habitats, is known to have toxic effects on nervous and muscle function in teleost fish. Golden gray mullet (Liza aurata L.) were exposed for 24 h to sublethal ammonia concentrations in seawater (control, 400 micromol L(-1), or 1,600 micromol L(-1) NH(4)Cl), and then their response to startling with a mechanical stimulus was measured with high-speed video. Initiation of the escape response was significantly slowed by ammonia exposure: response latency rose proportionally from <50 ms in controls to >300 ms at a concentration of 1,600 micromol L(-1 ) NH(4)Cl. This indicates toxic effects on nervous function within the reflex arc. Impaired escape performance was also observed: maximum turning rate, distance covered, velocity, and acceleration were significantly reduced by >45% at a concentration of 1,600 micromol L(-1) NH(4)Cl. This indicates toxic effects on fast-twitch glycolytic white muscle function, the muscle type that powers the fast-start response. These neuromotor impairments were associated with significant ammonia accumulations in venous plasma and white muscle and brain tissue. These results indicate that anthropogenic ammonia pollution in aquatic habitats may increase the vulnerability of fish to predation, especially by birds and mammals that are not affected by water ammonia concentrations.

Environmental hypoxia as a metabolic constraint on fish: the case of Atlantic cod, Gadus morhua.

Hypoxia is known to provoke a wide range of effects on aquatic animals. Here we use laboratory and field data on Atlantic cod, Gadus morhua, to illustrate that many of these responses can be explained within the metabolic scope (MS) framework, i.e. taking into account the directive and limiting effects of dissolved oxygen (DO) on the ability of animals to acquire energy for growth and activity. A MS model for cod shows that scope for activity (swimming, feeding, etc.) is proportional to DO and becomes nil, jeopardising survival, when DO is < approximately 20% air saturation. Laboratory studies have confirmed this lethal threshold and demonstrated that growth and food ingestion were significantly reduced below 70% sat. This loss of appetite has been linked to a reduction of the peak value and an increase in duration of postprandial metabolism, in agreement with the MS model. Dwindling MS during hypoxia imposes an upper limit to swimming performance. Cod may also opt to reduce spontaneous swimming activity to spare oxygen for other activities such as digestion. In the Kattegat, the Baltic Sea, and the Gulf of St. Lawrence, eastern Canada, cod completely avoid waters where their MS is near zero. Furthermore, cod density increases exponentially with DO up to approximately 70% sat in the Gulf of St. Lawrence. Although hypoxia results in other direct and indirect effects as well, the MS framework allows modelling of many of the responses to hypoxia for individual cod that ought to be reflected at the population and community levels. The MS framework is also useful to compare species responses. We show that the impact of hypoxia on MS is similar, when expressed as a proportion of MS in normoxia, in cod, European sea bass (Dicentrarchus labrax), the common sole (Solea solea) and turbot (Psetta maxima). Data are required for other species to evaluate how general these findings are.

Environmental constraints upon locomotion and predator-prey interactions in aquatic organisms: an introduction.

Environmental constraints in aquatic habitats have become topics of concern to both the scientific community and the public at large. In particular, coastal and freshwater habitats are subject to dramatic variability in various environmental factors, as a result of both natural and anthropogenic processes. The protection and sustainable management of all aquatic habitats requires greater understanding of how environmental constraints influence aquatic organisms. Locomotion and predator-prey interactions are intimately linked and fundamental to the survival of mobile aquatic organisms. This paper summarizes the main points from the review and research articles which comprise the theme issue 'Environmental constraints upon locomotion and predator-prey interactions in aquatic organisms'. The articles explore how natural and anthropogenic factors can constrain these two fundamental activities in a diverse range of organisms from phytoplankton to marine mammals. Some major environmental constraints derive from the intrinsic properties of the fluid and are mechanical in nature, such as viscosity and flow regime. Other constraints derive from direct effects of factors, such as temperature, oxygen content of the water or turbidity, upon the mechanisms underlying the performance of locomotion and predator-prey interactions. The effect of these factors on performance at the tissue and organ level is reflected in constraints upon performance of the whole organism. All these constraints can influence behaviour. Ultimately, they can have an impact on ecological performance. One issue that requires particular attention is how factors such as temperature and oxygen can exert different constraints on the physiology and behaviour of different taxa and the ecological implications of this. Given the multiplicity of constraints, the complexity of their interactions, and the variety of biological levels at which they can act, there is a clear need for integration between the fields of physiology, biomechanics, behaviour, ecology, biological modelling and evolution in both laboratory and field studies. For studies on animals in their natural environment, further technological advances are required to allow investigation of how the prevailing physico-chemical conditions influence basic physiological processes and behaviour.

Associations between tissue fatty acid composition and physiological traits of performance and metabolism in the seabass (Dicentrarchus labrax).

Seabass were fed for 4 months with diets where the lipid was provided as either canola oil (CO), palm oil (PO) or fish oil (FO), to generate diversity in their tissue fatty acid (FA) composition and investigate how this influenced major traits of exercise performance, cardiac performance and respiratory metabolism. In particular, based upon previous observations, we investigated the hypothesis that enriching the fish tissues with oleic and linoleic acids (OA, 18:1n-9 and LA, 18:2n-6, respectively) from the CO and PO diets would improve maximum exercise and cardiac performance, and increase aerobic metabolic scope. This proved to be the case; exercise respirometry on bass fitted with cardiac flow probes revealed that those fed CO and PO diets had a significantly higher critical swimming speed (U(crit)) than those fed the FO diet. The improved swimming performance in the CO and PO groups was accompanied by a higher maximum cardiac output (Q) and net cardiac scope, and a higher active metabolic rate (AMR) and aerobic scope (AS) than in the FO group. Analysis of tissue FA composition revealed that the fish fed the CO and PO diets had accumulated significantly higher levels of OA and LA in their heart and muscle than the fish from the FO group, which had significantly higher levels of highly unsaturated FA of the n-3 series, such as EPA and DHA (20:5n-3 and 22:6n-3, respectively). Principal components analysis revealed significant positive associations between tissue OA and LA content and U(crit), maximum Q, the increase in Q during exercise, AMR and aerobic scope. There was a negative association between these physiological traits and tissue content of EPA. Therefore, diet composition is an environmental factor that can generate significant phenotypic diversity in major physiological traits of performance and metabolism in the seabass, with increased intake of FAs such as OA and LA leading to improved cardiorespiratory performance.

Reflex cardioventilatory responses to hypoxia in the flathead gray mullet (Mugil cephalus) and their behavioral modulation by perceived threat of predation and water turbidity.

In hypoxia, gray mullet surface to ventilate well-oxygenated water in contact with air, an adaptive response known as aquatic surface respiration (ASR). Reflex control of ASR and its behavioral modulation by perceived threat of aerial predation and turbid water were studied on mullet in a partly sheltered aquarium with free surface access. Injections of sodium cyanide (NaCN) into either the bloodstream (internal) or ventilatory water stream (external) revealed that ASR, hypoxic bradycardia, and branchial hyperventilation were stimulated by chemoreceptors sensitive to both systemic and water O2 levels. Sight of a model avian predator elicited bradycardia and hypoventilation, a fear response that inhibited reflex hyperventilation following external NaCN. The time lag to initiation of ASR following NaCN increased, but response intensity (number of events, time at the surface) was unchanged. Mullet, however, modified their behavior to surface under shelter or near the aquarium edges. Turbid water abolished the fear response and effects of the predator on gill ventilation and timing of ASR following external NaCN, presumably because of reduced visibility. However, in turbidity, mullet consistently performed ASR under shelter or near the aquarium edges. These adaptive modulations of ASR behavior would allow mullet to retain advantages of the chemoreflex when threatened by avian predators or when unable to perceive potential threats in turbidity.

Physiology and behaviour of free-swimming Atlantic cod (Gadus morhua) facing fluctuating temperature conditions

Atlantic cod (Gadus morhua L.), acclimated to 5 °C, were equipped with ultrasonic transmitters which allowed the continuous monitoring of their vertical movements and heart rate. Fish were then placed in a 125 m3 tower tank in which the various thermal conditions they encounter in their natural environment were reproduced. Physiological and behavioural responses of cod were followed in parallel to the induced environmental changes. The experimental conditions studied in the tower tank were also reproduced in a swimming respirometer, where oxygen consumption and heart rate could be monitored within the activity range of a free-swimming animal. In a homogeneous water column, a rise in temperature induced marked increases in fish swimming activity, heart rate and heart beat-to-beat variability. In a thermally stratified environment, voluntary activity also increased when the thermal structure of the water column was altered, though no temperature-dependent changes in heart rate were observed. In this case, fish avoided the new temperature conditions, exhibiting distinct thermoregulatory behaviour. Stratification of the water column also prompted daily cyclic changes in fish distribution, animals tending to be in deeper and colder water layers during the day and in shallower and warmer layers at night. Respirometry experiments revealed that the thermoregulatory behaviour observed in free-ranging fish was probably driven by the energetic expedient of maintaining the physiological status quo ­ i.e. avoiding bioenergically costly reacclimation processes. Indeed, acute temperature increases or decreases of 2.5 °C led to marked differences in oxygen consumption, with metabolic rate changes of 15 and 30 %, respectively. The persistent linear relationship between heart rate and oxygen consumption allowed us to estimate, from the heart rate recorded in free-swimming fish, the entire range of metabolic responses that cod underwent voluntarily while experiencing a thermally stratified water column. The most profound metabolic effect, however, was observed with feeding, when oxygen consumption increased by as much as 80 %, resulting in an estimated 90 % reduction in their subsequent scope for activity.

Physiology and behaviour of free-swimming Atlantic cod (Gadus morhua) facing fluctuating salinity and oxygenation conditions

1. Atlantic cod (Gadus morhua L.) acclimated to a temperature of 5 °C and 30 salinity were equipped with ultrasonic transmitters which allowed continuous monitoring of their heart rate and their position in the water column. Fish were placed in a 125 m3 tower tank which permitted various environmentally relevant modifications of the salinity and oxygenation conditions. Cod physiological and behavioural responses were followed in parallel to the environmental manipulations. Some of the experimental conditions studied in the tower tank were also reproduced in a swimming respirometer where fish oxygen consumption and heart rate were monitored at various levels of activity. 2. Lowering salinity from 30 to 26 did not change resting oxygen consumption, but increased active oxygen consumption. 3. Lowering salinity from 30 to 26 increased heart rate over the whole range of swimming speeds except at maximum speed. 4. Lowering oxygen tension to 9 kPa decreased oxygen consumption over the whole range of swimming speeds and decreased resting heart rate. 5. Low salinity did not significantly affect the relationship between heart rate and oxygen consumption. 6. Low oxygen levels decreased the oxygen transported per heart beat. 7. In the tower tank, bursts of activity were associated with tachycardias. 8. In uniform conditions, fish swam more deeply during the day than at night. 9. After an exploratory period of approximately 6 h, fish chose to remain in a low-salinity upper layer of the tank. Thereafter, high salinities were avoided. Fish tended to select low salinities if a choice was provided. 10. Fish generally avoided zones of low oxygen (<9 kPa) but continued voluntarily to enter regions with values as low as 3.0 kPa for short excursions or if food was offered.

Adaptive respiratory responses of trout to acute hypoxia. II. Blood oxygen carrying properties during hypoxia.

The effects of deep and acute hypoxia (PwO2 = 25 Torr) on oxygen transport characteristics (Hill number (n) and P50) were investigated in trout at 15 degrees C. When a fish is submitted to such an acute and deep hypoxia, a metabolic acidosis develops as soon as the arterial oxygen tension drops to about 15 Torr. We first showed that the hemoglobin of blood sampled at the end of the acidification period has an increased oxygen affinity. This improved affinity could be explained by the internal alkalisation of erythrocytes due to the extrusion of protons via a beta-adrenergic stimulation of Na+/H+ exchanges occurring at the onset of hypoxia and responsible for extracellular acidosis. Secondly we observed a significant increase (about 20%) of the number of blood cells per volume of blood during the acidosis. This cell number stays constant afterwards. The dual effects of a higher hemoglobin oxygen affinity and a greater amount of available hemoglobin improving blood oxygen loading at the fish gills appear to be a fast adaptive response to acute hypoxia. Surprisingly, we found that the elevated affinity occurring during acidosis remained constant as long as the fish were maintained in hypoxia, in spite of possible large variations of extracellular pH (pHe). This result is difficult to reconcile with the idea that the increase in affinity is imposed by intracellular pH (pHi), since in red blood cells pHi depends on pHe, thus any modification of pHe would in this case modify oxygen affinity.

Adaptive respiratory responses of trout to acute hypoxia. III. Ion movements and pH changes in the red blood cell.

In the preceding paper acute hypoxia was shown to elicit within minutes an increase in the blood O2 affinity. From the present data it appears that this rapid change in blood P50 value can be ascribed to an important alkalization of the red blood cell despite a simultaneous decrease in extracellular pH (pHe). The intracellular alkalization is only partially due to beta-adrenergic stimulation of Na/H exchange, deoxygenation of hemoglobin and the rapid decrease of PaCO2 due to hyperventilation being involved in this process via the chloride shift. This high value of intraerythrocytic pH (pHi) is then maintained practically constant throughout the time the fish is kept in hypoxia despite wide changes of external pH. The blocking of pHi accounted for the constant O2 content observed during hypoxia. The uncoupling of pHi from pHe, which occurs at the onset of hypoxia, is still unexplained: for instance, it is not due to inhibition of the anion exchanger responsible for the passive distribution of H+ across the red cell membrane. A general scheme of all the mechanisms involved in the emergency adaptive response to acute hypoxia is presented.

Adaptive respiratory responses of trout to acute hypoxia. I. Effects of water ionic composition on blood acid-base status response and gill morphology.

The effects of various levels of hypoxia (PWO2 ranging from 10 to 60 Torr) on arterial blood gases (PaO2 and PaCO2) and acid-base status were investigated in trout at 15 degrees C. The hypoxic responses of two stocks of trout living in natural waters having very different levels of NaCl (1.0 mmol.L-1 and 0.1 mmol.L-1) and carbonate alkalinity (0.4 mmol.L-1 and 2.4 mmol.L-1) were compared. The use of an extracorporeal circulation method made it possible to continuously monitor the pH changes. The different patterns of the acid-base status observed in response to hypoxia depend on the evolution of PaO2. Two critical PaO2 thresholds were defined. Crossing the upper (about 15 Torr) induces metabolic acidosis which is normally followed by pH recovery, while crossing the lower (about 10 Torr) promotes loss of capacity to compensate acidosis. The NaCl concentration of the water drastically modifies the fish sensitivity to hypoxia: fish living in water with a low NaCl concentration have less resistance to hypoxic exposure. This may be explained by the fact that in fish living in low NaCl concentrations, the secondary gill lamellae are surrounded by chloride cells, which considerably reduce the surface area available for gas exchange. Consequently a modest fall in PWO2 induces a drastic reduction of the arterial oxygen tension which crosses the lower critical PaO2 threshold.