Sea bass is a major species in Mediterranean aquaculture, and is now being subject to selective breeding programmes for faster growth. In terrestrial species, it was demonstrated that fast growth may be linked to a correlated degradation of fitness traits. In this experiment, we evaluated 600 young sea bass from a factorial mating of 76 sires and 13 dams. The sires were from four genetic groups, wild (W), domesticated (D), and selected for growth (2 groups, M and P). The 600 offspring were submitted to two acute confinement stress challenges at 6 weeks intervals, and plasma cortisol at one hour post stress was measured. The same fish were also submitted to two swimming challenges at a 5 days interval, where the maximum sustained swimming speed (U-max) of each fish was evaluated. Parentage was assessed by genotyping of 12 microsatellites. 554 fish had both valid parentage and phenotypes. Cortisol had a low repeatability (r = 0.30 between the two successive measurements) while repeatability was moderate for U-max (r = 0.62). However, genetic correlations between successive measurements were very high (>0.96) for both traits, indicating that successive measurements were related to the same trait. Heritability was moderate for mean post-stress cortisol (h(2) = 0.34 +/- 0.09) and U-max (h(2) = 0.48 +/- 0.08). When U-max was expressed in m.s(-1), it was negatively correlated to cortisol (r(A) = -0.48 +/- 0.08) and weakly correlated to body weight (r(A) = 0.12 +/- 0.16), but figures changed when it was expressed in Body Lengths.s(-1) (h(2) = 0.55 +/- 0.08, r(A) = -0.10 +/- 0.19 with cortisol and r(A) = -0.64 +/- 0.07 with body weight, respectively). Cortisol was moderately negatively correlated with body weight (r(A) = -0.36 +/- 0.18). The four lines did not differ for cortisol or U-max, but when U-max was expressed in BL.s(-1) it tended to be lower in the two selected lines - which were also significantly larger. However, this is likely due to a phenotypic decrease of relative U-max with increasing body size. We conclude that selection for growth and/or domestication should not impact maximum sustained swimming speed in the European sea bass, but may tend to favour animals with low cortisol responsiveness. These traits could be used to orientate functional capabilities other than productivity in sea bass.Statement of relevance: We estimate heritability of cortisol stress response and (for the first time) in European sea bass, as well as their correlations with growth. We show moderate correlations, and no correlated response to selection for growth. We also provide methods to evaluate these traits on large number of fishes. This can be useful to monitor and design breeding programmes. (C) 2016 Elsevier B.V. All rights reserved.

In this study, the consequences of the exposure of rainbow trout (Oncorhynchus mykiss) to a brief stress during early development were explored on the later response of fingerlings to stress. Firstly, we analyzed the ontogeny of cortisol production and that of the initial cortisol response to stress in developing fish. It is only at the eyed stage that the embryos started to produce some basal cortisol. The HPI (hypothalamic-pituitary-interrenal axis) was however not functional before hatching, as exposure of the embryos to a stress did not trigger any cortisol response. A cortisol response to an acute stress was detected 9 days after hatching. In a second set of experiments, we showed that a very brief stress applied at 3 different early stages (eyed, hatching, and yolk resorption) reduced the later cortisol response to stress of 5-month-old fingerlings. This reduction is not likely to be due to alterations in the fish interrenal sensitivity because the 5-month-old fingerlings responded to ACTH treatment (only one dose tested) within the same magnitude as the fish that were not stressed during early development. An experimentally induced increase in egg cortisol just after fertilization also induced a reduction in stress sensitivity of 5 month old fingerlings, which was dose-dependent This study shows for the first time that the responsiveness of the corticotrope axis in 5-6 months old rainbow trout was influenced both by early stress exposure and by initial egg cortisol levels. Whether the HPI was functional or not at the time the initial stress was applied only had a small influence on the later unambiguous effect of early stress exposure.

The present paper describes the main procedures used to slaughter fowl, pigs, calves and adult cattle, sheep, and farmed fish, starting on the farm and ending with the death of the animal at the abattoir. It reviews the currently known causes of stress, indicated by behavioural and physiological measurements on the animal level, and by post-mortem muscle metabolism. During the pre-slaughter period, psychological stress is due to changes of environment, social disturbances and handling, and physical stress is due to food deprivation, climatic conditions, fatigue, and sometimes pain. The exact causes of stress depend, however, on the characteristics of each species, including the rearing system. For fowl, bird catching and crating, duration and climatic conditions of transport and of lairage and shackling are the main known pre-slaughter stress factors. For pigs, stress is caused by fighting during mixing of pens, loading and unloading conditions, and introduction in the restrainer. Handling and novelty of the situation contribute to the stress reactions. For veal calves and adult cattle, disruption of the social group, handling, loading and sometimes unloading conditions, fatigue, novelty of the situation and for calves mixing with unfamiliar animals are known stress factors. Gathering and yarding of extensively reared lambs and sheep causes stress, particularly when shepherd dogs are used. Subsequent transport may induce fatigue, especially if sheep are commercialised through auctions or markets. In farmed fish, stress is predominantly related to environmental aspects such as temperature, oxygen, cleanliness of the water and, to a certain extent, stocking density and removal of the fish from the water. If transport and lairage conditions are good and their durations not too long, they may allow pigs, calves and adult cattle, sheep, and fish to rest. For certain species, it was shown that genetic origin and earlier experience influence reactions to the slaughter procedure. Stunning techniques used depend on the species. Pigs and fowl are mostly electrically or gas-stunned, while most adult cattle are stunned with a captive bolt pistol. Calves and sheep may be electrically stunned or with a captive bolt pistol. Various stunning methods exist for the different farmed fish species. Potential causes of stress associated with the different stunning procedures are discussed. The paper addresses further consequences for meat quality and possible itineraries for future research. For all species, and most urgently for fish, more knowledge is needed on stunning and killing techniques, including gas-stunning techniques, to protect welfare.

Although effects of hypoxia and, in a lower extent, effects of hyperoxia, have been widely studied, very few data is available regarding the effect of oxygen concentration around normoxia. Moreover, the effect of rearing oxygen level around normoxia on flesh quality, and a possible interaction with stress at slaughter, have never been investigated. Rainbow trout were reared during 18 weeks up to pan size (mean weight 400 g) at 76, 98 or 117% of oxygen saturation. Afterwards they were slaughtered according to two procedures: a minimal stress (NS group), or after a 15 min confinement stress (S group). Fish characteristics (weight, condition factor, fat-meter value, plasma cortisol level, hepato-somatic index, and hepatic glycogen content) and quality criteria of fillets (color, initial muscle pH, and muscle glycolytic potential) were recorded at slaughter time, whereas mechanical resistance, ultimate pH and dry matter content of the flesh were measured at 24 h and 48 h post-mortem. Muscle fiber size was analyzed on histological samples from white and red muscles. In our study, rearing oxygen level had no impact on the morphological characteristics of the fish except a higher hepato-somatic index and hepatic glycogen content for trout reared at 97% oxygen saturation. Rearing oxygen level affects slightly flesh quality at the lowest oxygen level (76% saturation) with a lower force for low deformation mechanical resistance measurement in the anterior part of the fillet. Stress before slaughter highly increased plasma cortisol concentration (by more than 18 in mean), and decreased fat-meter values (of 14% in mean) and the mean diameter of red muscle fibers (of 6.4% in mean). Stress before slaughter also decreased fillet lightness (L*, -1.1 unit), yellowness (b*, -0.76 unit), and initial pH (-0.15 unit) of the flesh but had no effect on ultimate pH measured at 24 or 48 h post-mortem. Stress before slaughter induced a softer flesh as shown by lower mechanical resistance at high deformation measured on samples from both anterior and caudal part of the fillet. The post-mortem measurement time (24 or 48 h) was the main factor affecting the mechanical resistance of the flesh, with a significant softening of the flesh between 24 and 48 h. Nor oxygen level, nor slaughter stress affected the size of white muscle fibers. No major interaction between rearing oxygen level and slaughter stress was formally demonstrated. (C) 2008 Elsevier B.V. All rights reserved.

La multiplicité des critères susceptibles d’être utilisés pour évaluer l’adaptation comportementale d’un animal à son environnement a motivé l’étude des relations entre réponses physiologiques et comportementales, dans l’espoir que l’identification de telles relations puisse simplifier l’évaluation du bien-être animal. La comparaison d’animaux placés dans des environnements différents et la comparaison d’animaux aux génotypes extrêmes suggèrent l’existence de relations entre certaines réponses physiologiques et comportementales. La possibilité de catégoriser certains animaux selon leur stratégie de réponse adaptative, caractérisée par des associations répétables entre réponses comportementales et physiologiques, conforte cette hypothèse. Cependant, les relations ainsi suggérées semblent difficilement généralisables : il n’existe pas de relation univoque entre les réponses physiologiques et comportementales d’un animal face à une situation perçue comme menaçante. En l’état actuel des connaissances, la prise en compte de manière concomitante de critères physiologiques et comportementaux est essentielle pour apprécier le bien-être d’un animal. A l’avenir, des investigations en génomique fonctionnelle et en neurobiologie pourraient améliorer notre compréhension de ces relations.

Malgré les progrès de ces dernières années, la période de pré-abattage est une source de stress pour les animaux. Les procédés d’abattage nécessitent des regroupements et des mélanges d’animaux, l’enlèvement du milieu habituel et l’introduction dans des environnements non familiers, le transport, la manipulation par l’homme et le jeûne, et entraînent parfois des mauvaises conditions d’ambiance. Ils sont souvent générateurs de stress d’origine physique (fatigue, faim, douleur, inconfort physique) et psychologique (peur, stress social). Les techniques d’étourdissement sont parfois mal maîtrisées ou mal adaptées. Ces procédés entraînent des réponses comportementales, physiologiques et métaboliques qui sont utilisées pour évaluer le niveau de stress de l’animal. Afin de mieux respecter le bien-être animal à l’abattage, il est nécessaire d’élargir nos connaissances des causes de stress. L’effet négatif que peuvent avoir ces réponses sur les qualités des viandes est succinctement abordé.

Despite progress made in recent years, the pre-slaughter period remains stressful for animals. Current slaughter procedures involve regrouping and mixing animals, removing them from their familiar environment and introducing them into unfamiliar settings, transporting, handling, and depriving them of food, and are sometimes associated with poor quality of surroundings. These often cause stress, which can be both physical (fatigue, hunger, pain, discomfort) and psychological (fear, social stress). Stunning techniques are sometimes poorly controlled or unsuitable. Slaughter procedures cause behavioural, physiological and metabolic responses that are used to determine the animal's stress levels. To improve animal welfare, further knowledge of the exact causes of stress is required. The possible negative effects of stress responses on meat quality are also briefly addressed.

The recent development of aquaculture has raised interest in fish welfare. It is debatable whether fish can feel pain, although there is a certain consensus that fish can perceive harmful stimuli. However, it is still difficult to evaluate how this information is integrated as a subjective experience. When exposed to stress, fish develop the same physiological and behavioural responses as terrestrial vertebrates. Analysing this information using a multiparameter approach has enabled fish welfare to be defined. Several activities involved in fish farming put fish welfare at risk. Deterioration of water quality can be a major cause of welfare problems with regards to specific characteristics of the species. Other parameters such as fish density, handling and transport are also likely to impact on welfare. Control of these stress factors by fish farmers is essential for successful production. In addition to experience acquired by fish farmers, better knowledge of the biological basis of fish welfare is required.

Assessing the behavioural adaptation of an animal to its environment is complex, notably because numerous criteria can be taken into consideration. A better understanding of the relationships between criteria, particularly between behavioural and physiological responses, might help reduce the number of parameters required to assess animal welfare. The existence of relationships between behavioural and physiological responses of animals to potentially threatening situations has been suggested both by studies comparing animals reared in different environments and by those comparing animals with extreme genotypes. Moreover, the identification of coping styles in various species has strengthened the idea that physiological and behavioural responses could be related, although the precise laws governing such relationships are still difficult to establish and generalise. Thus, considering the complexity of these relationships, it appears reasonable to consider these two groups of responses as partially independent and as giving complementary information about animal welfare. In the future, the development of complex trait analyses as well as investigation at the level of the brain should improve understanding of the relationships between physiological and behavioural responses.

Le récent développement de l’aquaculture a conduit à s’intéresser au bien-être des poissons en élevage. A la question du ressenti de la douleur chez le poisson, il est le plus souvent admis que ces animaux sont capables de la percevoir mais pas nécessairement d’en avoir la même conscience au niveau cérébral que chez l’homme. Exposés à des situations de stress, les poissons présentent globalement les mêmes stratégies physiologiques et comportementales que les vertébrés terrestres. C’est à partir de ces informations qu’a été abordée la caractérisation du bien-être chez les poissons en favorisant une approche multiparamétrique. Les activités associées à l’élevage des poissons constituent autant de facteurs susceptibles de modifier leur niveau de bien-être. La qualité physico-chimique du milieu d’élevage en rapport avec les spécificités de l’espèce élevée constitue une cause importante de modification du bien-être. D’autres facteurs comme la densité d’élevage, la manipulation du poisson ou son transport sont également susceptibles d’avoir un impact sur le bien-être. Le contrôle de tous ces facteurs par l’éleveur est indispensable pour la réussite d’un élevage. Les compétences acquises par les éleveurs doivent être complétées par une meilleure connaissance des bases biologiques du bien-être chez les poissons.

Evaluating the ecological impact of an oil spill is a complex issue requiring coherently articulated examination of the sequence of interactions that link the cell, where contaminants exert their effects, to the ecosystem, where interactions with human activities arise. This sequence of interactions traverses the frontiers between scientific disciplines (chemistry, toxicology, physiology, and fisheries ecology). Using the common sole (Solea solea L.) as a model species for the coastal habitats polluted by the "Erika" oil spill, our research project attempted to define indices of functional integrity that characterised the consequences of fuel exposure at the different biological levels. The coupling of field observations with experimental laboratory work revealed how functional alterations which are readily observable within individuals and their organs are progressively obscured as investigation progresses towards more complex organisational levels. Some of the approaches and indices are proposed as instruments for evaluating the impact of contamination by hydrocarbons.

The effects of chronic confinement stress (1, 5 and 10 days) and of periodic blood sampling on somatic growth and the structure and growth of otoliths was studied in Oreochromis niloticus. During the study, the plasma concentrations of cortisol were measured at various times during the application of stress: they were significantly higher in confined fish than in control fish (mean ± s.d. 3·40 ± 0·47 v. 1·26 ± 0·62 ng ml−1, P < 0·05) up to 5 days after the start of a 10 day stress period. The somatic growth (standard length, LS, and mass) was affected by the confinement and by the sampling (from 16·21 ± 1·07 to 14·64 ± 1·15 cm for LS, and from 173·31 ± 33·14 to 110·50 ± 29·48 g for mass). But the confinement masked the effect of the sampling on somatic growth. Tetracycline was injected at the start of the experiment to mark the otoliths, and showed that the short and long duration confinements led to a clear check in the pattern of primary increments in the otoliths. The number of primary increments deposited during the resting periods that followed each period of confinement was always less than the number of days that these periods lasted. No relation was found between the duration of confinement and the structure of the resulting checks. These results suggest that there is a disruption in the laying down of primary increments during periods of confinement resulting in an underestimation of their number compared to the actual number of days of growth. These results call into question the use of otolith primary increments as a means of estimating the age of Nile tilapia that have experienced periods of stress.

In this study, the expression of several genes involved in cortisol synthesis in head kidneys, the site of cortisol production, and in the rainbow trout (Oncorhynchus mykiss) was examined in response to two different acute stressors and an acute ACTH treatment. mRNAs levels of the "steroidogenic acute regulatory" (StAR) sterol transport protein, which transports cholesterol to the inner mitochondrial membrane as well as cytochrome P450 cholesterol side chain cleavage (P450(SCC)) were determined in head kidney (containing the interrenal tissue). In one experiment, we also quantified 3-beta-hydroxysteroid dehydrogenase (3B-HSD) and cytochrome P450(11beta) (11B-H) mRNAs. The presence of these four transcripts in the head kidney was confirmed by Northern blot analysis. For each stress condition, mRNA levels were quantified by quantitative or real-time RT-PCR. The results of these two methods were highly correlated. An acute stress induced by capture, short confinement (2min), and anesthesia (3min) resulted in significant elevation of plasma cortisol (30-fold higher than controls) and an increase in levels of StAR and P450(SCC) mRNAs 3h post-stress. When fish were submitted to an acute stress caused by 5min of chase with a net in a tank, plasma cortisol reached a peak within 1h, but after 3h, levels were only 5-fold higher in stressed trout than in controls and no variations in the expression of StAR, P450(SCC), 3B-HSD, and 11B-H were observed whatever the time post-stress. One hour after acute ACTH stimulation (5IU/kg), plasma cortisol level was 4-fold higher than in control trout and no changes in StAR and P450(SCC) mRNAs levels were detected. The data suggest that the high levels of cortisol after stress need an activation of genes involved in cortisol synthesis, but lower levels do not. Futhermore, under these three test conditions, we always found a strong positive correlation between mRNA levels of StAR and P450(SCC), in contrast to what has been described in mammals. Consequently, the absence of transcription activation with low increase in cortisol levels suggests that other levels of regulation, particularly activation of pre-existing proteins, govern cortisol production.

The effects of concentration (5,50, and 500 μg/L) and duration (24, 48 h) of exposure to carbofuran, a carbamate insecticide, were assessed on brain catecholamine (norepinephrine [NE] and dopamine), plasma glucose, and hepatic glycogen contents and behavioral activities of goldfish ( Carassius auratus ). After 24 h of exposure to 50 and 500 μg/L, the level of NE was increased in the olfactory bulbs. The same effect was observed after a 48‐h exposure to 500 and 50 μg/L in the telencephalic hemispheres and in the hypothalamus, respectively. An increase in the level of dopamine was also found in hypothalamus after 48 h of exposure to 500 μg/L carbofuran. Plasma glucose increased in concentration after both periods of exposure to carbofuran at 50 and 500 μg/L. Hepatic glycogen concentration decreased after a 48‐h exposure to the highest concentration. Behavioral endpoints related to swimming pattern and social interactions were affected after a 24‐h exposure to the lowest concentration tested (5 μg/L). The relative sensitivities of these different types of responses to exposure to carbofuran are discussed in light of data on the neurotoxic effects of carbamate and organophosphate insecticides in fish.

The aim of the present study was to assess a potential link between confinement stress and prolactin (PRL), the hormone responsible for adaptation to a hypoosmotic environment in freshwater-adapted tilapia (Oreochromis niloticus). The effect of stress on plasma levels of the two tilapia PRL forms, tiPRLI (or tiPRL188) and tiPRLII (or tiPRL177), was examined along with the effects on plasma levels of cortisol and growth hormone (GH). In a preliminary study, various sampling protocols (immediate sampling; sampling one by one; anesthesia at 0.5, 1, 2 ml/liter phenoxyethanol) were tested for their ability to modify basal plasma PRL and cortisol. In fish sampled within 1 min of capture (immediate sampling), no changes in the plasma levels of these hormones were observed, whereas when fish were sampled one at a time, PRL levels did not change but cortisol levels were modified. The immediate sampling protocol was used to study the effects of 1 hr confinement stress, which induced a large increase in plasma cortisol levels as well as increases tiPRLI and tiPRLII levels with kinetics similar to those of cortisol. In contrast, plasma tiGH levels significantly decreased after 1 hr confinement. When this stress situation was removed, plasma cortisol and tiPRL levels decreased and plasma GH levels increased. Two and one-half hours later, values were not significantly different from those measured in control fish. In tilapia exposed to 24 hr confinement stress, similar changes in hormone levels were observed. However, after 24 hr confinement, only cortisol levels were significantly different from those measured in control fish. None of these stress conditions significantly changed plasma chloride levels. Together, these results indicate that both PRL and GH have important roles in the adaptive response of freshwater-adapted tilapia to confinement stress.

BACKGROUND:One of the characteristic features of the two types (alpha and beta) of "mitochondria-rich" (chloride) cells in the gill epithelium of freshwater fishes is the presence in their apical region of tubulovesicular structures. A further analysis of the ultrastructural features of these apical elements as well as that of their modifications under various living conditions should help to understand better the respective rôle of both alpha and beta cells in these conditions.METHODS:Atlantic salmon (Salmo salar) maintained in fresh water as well as tilapia (Oreochromis niloticus) maintained either in fresh water or in deionized water or in 20% saltwater were examined. Measurements of surface areas of apical structures in the various living conditions were also performed.RESULTS:In the alpha cells of freshwater fishes, the apical structures consisted of isolated vesicles containing a filamentous material resembling that coating the apical surface. They were closely related to the apical plasma membrane and did not penetrate the region containing the tubular system. When fishes were transferred to deionized water, the number of the apical membrane folds increased significantly, as did the number and size of apical structures which became elongated. In saltwater-adapted fishes, the apical structures showed a tendency to collapse and took the appearance of flattened and slightly curved elements. These observations tended to indicate that in alpha cells the apical structures were extensions of the apical plasma membrane and thereby might be implicated in sodium uptake when fishes are placed in fresh or deionized water and in chloride excretion when they are transferred to salt water. In beta cells, the apical structures were usually separated from the apical plasma membrane by a zone rich in cytoskeleton elements. They penetrated deeply into the supranuclear region, where they intermingled with the elements of the tubular system. They consisted mainly of tubular elements that contained a material resembling that present in the trans tubular Golgi network from which they might originate. The apical structures remained unaltered in beta cells whatever the medium (fresh or deionized water) in which the fish was placed.CONCLUSIONS:The alpha cells which are usually thought to be mainly involved in chloride excretion when fishes are transferred into seawater might also be implicated in sodium uptake in freshwater living conditions. The rôle of beta cells, in contrast, still remains to be established.