Minding the Gaps in Fish Welfare: The Untapped Potential of Fish Farm Workers.

The welfare of farmed fish is often regarded with less concern than the welfare of other husbandry animals, as fish are not universally classified as sentient beings. In Norway, farmed fish and other husbandry animals are legally protected under the same laws. Additionally, the legislature has defined a number of aquaculture-specific amendments, including mandatory welfare courses for fish farmers who have a key role in securing animal welfare, also with regards to noting welfare challenges in the production process. This article uses fish welfare courses as a site from which to inquire about the common-sense understanding of fish welfare in Norwegian fish farming. The focus is specifically on fish farm employees, their experiences of welfare-related issues and contradictions in their daily work, and the struggle to act responsibly in aquaculture settings. Through participant observation at welfare courses, as well as interviews and conversations with fish farm workers, the article details how challenges are experienced 'on the ground', and suggests how fish farm workers' own experiential knowledge might be mobilized to improve the general welfare of farmed fish.

Computational animal welfare: towards cognitive architecture models of animal sentience, emotion and wellbeing.

To understand animal wellbeing, we need to consider subjective phenomena and sentience. This is challenging, since these properties are private and cannot be observed directly. Certain motivations, emotions and related internal states can be inferred in animals through experiments that involve choice, learning, generalization and decision-making. Yet, even though there is significant progress in elucidating the neurobiology of human consciousness, animal consciousness is still a mystery. We propose that computational animal welfare science emerges at the intersection of animal behaviour, welfare and computational cognition. By using ideas from cognitive science, we develop a functional and generic definition of subjective phenomena as any process or state of the organism that exists from the first-person perspective and cannot be isolated from the animal subject. We then outline a general cognitive architecture to model simple forms of subjective processes and sentience. This includes evolutionary adaptation which contains top-down attention modulation, predictive processing and subjective simulation by re-entrant (recursive) computations. Thereafter, we show how this approach uses major characteristics of the subjective experience: elementary self-awareness, global workspace and qualia with unity and continuity. This provides a formal framework for process-based modelling of animal needs, subjective states, sentience and wellbeing.

Uncontrollable chronic stress reduces growth disparities in farmed Atlantic salmon.

Individual variation in behavior and physiological traits in a wide variety of animals has been the focus of numerous studies in recent years. In this context, early life experiences shape responses that individuals have to subsequent environments, i.e. developmental plasticity. In this experiment, we subjected 10-month old fish to an unpredictable chronic stress (UCS) regime or no stress (control) for 3weeks. These individuals then underwent the parr-smolt transformation, when salmonids become adapted for the seawater environment, and were subsequently transferred into seawater before the final sampling. Biometric data was collected at the end of each period. Sampling on the final day was conducted in order to analyze basal monoaminergic activity in the brain stem and hypothalamus, as well as gene expression of target genes in the telencephalon. We found that post-hoc sorting of individuals by their serotonergic activity (high and low) resulted in the elucidation of growth and gene expression differences. UCS groups were found to have less growth disparities throughout the experiment, compared to control fish. Furthermore, we found brain serotonergic signaling and corticotropic releasing factor binding protein expression were positively associated with brain stem serotonergic activity, which is consistent with fish showing a stress reactivity neurophysiological profile. In conclusion, we here submit evidence that sorting individuals by their basal serotonergic activity levels may be a useful tool in the study of developmental plasticity. These results may thus apply directly to improving husbandry practices in aquaculture and elucidating neural mechanisms for coping behavior.

Coping with a changing environment: the effects of early life stress.

Ongoing rapid domestication of Atlantic salmon implies that individuals are subjected to evolutionarily novel stressors encountered under conditions of artificial rearing, requiring new levels and directions of flexibility in physiological and behavioural coping mechanisms. Phenotypic plasticity to environmental changes is particularly evident at early life stages. We investigated the performance of salmon, previously subjected to an unpredictable chronic stress (UCS) treatment at an early age (10 month old parr), over several months and life stages. The UCS fish showed overall higher specific growth rates compared with unstressed controls after smoltification, a particularly challenging life stage, and after seawater transfer. Furthermore, subjecting fish to acute stress at the end of the experiment, we found that UCS groups had an overall lower hypothalamic catecholaminergic and brain stem serotonergic response to stress compared with control groups. In addition, serotonergic activity was negatively correlated with final growth rates, which implies that serotonin responsive individuals have growth disadvantages. Altogether, our results may imply that a subdued monoaminergic response in stressful farming environments may be beneficial, because in such situations individuals may be able to reallocate energy from stress responses into other life processes, such as growth.

Brain serotonergic activation in growth-stunted farmed salmon: adaption versus pathology.

Signalling systems activated under stress are highly conserved, suggesting adaptive effects of their function. Pathologies arising from continued activation of such systems may represent a mismatch between evolutionary programming and current environments. Here, we use Atlantic salmon (Salmo salar) in aquaculture as a model to explore this stance of evolutionary-based medicine, for which empirical evidence has been lacking. Growth-stunted (GS) farmed fish were characterized by elevated brain serotonergic activation, increased cortisol production and behavioural inhibition. We make the novel observation that the serotonergic system in GS fish is unresponsive to additional stressors, yet a cortisol response is maintained. The inability of the serotonergic system to respond to additional stress, while a cortisol response is present, probably leads to both imbalance in energy metabolism and attenuated neural plasticity. Hence, we propose that serotonin-mediated behavioural inhibition may have evolved in vertebrates to minimize stress exposure in vulnerable individuals.

Effect of predictability on the stress response to chasing in Atlantic salmon (Salmo salar L.) parr.

The possibility to prepare for and respond to challenges in a proper manner is essential to cope with a changing environment, and learning allows fish to up or downregulate the stress response based on experience. The regulation of the response to predicted needs should be easier in more predictable environments. We exposed salmon parr to chasing of either 15 s (weak stressor) or 5 min (strong stressor) twice daily for a 7-day learning period, with chasing either announced by a 30 s light signal (conditioned) or not announced (unconditioned). The behavioural response to the light signal was different between the conditioned and unconditioned groups, demonstrating that conditioned groups associated the signal with chasing. We could, however, not demonstrate any effect on the stress response of anticipation. The fish habituated to repeated stress exposures with a similar decrease in oxygen hyperconsumption in all groups. Due to habituation, possible effects of predictable announcement of a stressor on the physiological stress response may not have been expressed in this study. Plasma cortisol concentrations 1h after light signal and chasing the day after the training period was moderate in all groups although higher after 5 min chasing (13 ng ml(-1)) than 15 s chasing (7 ng ml(-1)). There was no physiological stress response after exposure to the light signal only after the learning period. We argue that the benefit of predictability of stressors is limited when the fish have no way to avoid the stressor.

A comparative study of the response to repeated chasing stress in Atlantic salmon (Salmo salar L.) parr and post-smolts.

When Atlantic salmon parr migrate from fresh water towards the sea, they undergo extensive morphological, neural, physiological and behavioural changes. Such changes have the potential to affect their responsiveness to various environmental factors that impose stress. In this study we compared the stress responses in parr and post-smolt salmon following exposure to repeated chasing stress (RCS) for three weeks. At the end of this period, all fish were challenged with a novel stressor and sampled before (T0) and after 1h (T1). Parr had a higher growth rate than post-smolts. Plasma cortisol declined in the RCS groups within the first week suggesting a rapid habituation/desensitisation of the endocrine stress axis. As a result of the desensitised HPI axis, RCS groups showed a reduced cortisol response when exposed to the novel stressor. In preoptic area (POA) crf mRNA levels were higher in all post-smolt groups compared to parr. 11ßhsd2 decreased by RCS and by the novel stressor in post-smolt controls (T1), whereas no effect of either stress was seen in parr. The grs were low in all groups except for parr controls. In pituitary, parr controls had higher levels of crf1r mRNA than the other parr and post-smolt groups, whilst pomcb was higher in post-smolt control groups. Overall, 11ßhsd2 transcript abundance in parr was lower than post-smolt groups; after the novel stressor pomcs, grs and mr were up-regulated in parr control (T1). In summary, we highlight differences in the central stress response between parr and post-smolt salmon following RCS.

Stress in Atlantic salmon: response to unpredictable chronic stress.

Combinations of stressors occur regularly throughout an animal's life, especially in agriculture and aquaculture settings. If an animal fails to acclimate to these stressors, stress becomes chronic, and a condition of allostatic overload arises with negative results for animal welfare. In the current study, we describe effects of exposing Atlantic salmon parr to an unpredictable chronic stressor (UCS) paradigm for 3 weeks. The paradigm involves exposure of fish to seven unpredictable stressors three times a day. At the end of the trial, experimental and control fish were challenged with yet another novel stressor and sampled before and 1 h after that challenge. Plasma cortisol decreased steadily over time in stressed fish, indicative of exhaustion of the endocrine stress axis. This was confirmed by a lower cortisol response to the novel stressor at the end of the stress period in chronically stressed fish compared with the control group. In the preoptic area (POA) and pituitary gland, chronic stress resulted in decreased gene expression of 11ßhsd2, gr1 and gr2 in the POA and increased expression of those genes in the pituitary gland. POA crf expression and pituitary expression of pomcs and mr increased, whereas interrenal gene expression was unaffected. Exposure to the novel stressor had no effect on POA and interrenal gene expression. In the pituitary, crfr1, pomcs, 11ßhsd2, grs and mr were down-regulated. In summary, our results provide a novel overview of the dynamic changes that occur at every level of the hypothalamic-pituitary gland-interrenal gland (HPI) axis as a result of chronic stress in Atlantic salmon.

Coping with unpredictability: dopaminergic and neurotrophic responses to omission of expected reward in Atlantic salmon (Salmo salar L.).

Comparative studies are imperative for understanding the evolution of adaptive neurobiological processes such as neural plasticity, cognition, and emotion. Previously we have reported that prolonged omission of expected rewards (OER, or 'frustrative nonreward') causes increased aggression in Atlantic salmon (Salmo salar). Here we report changes in brain monoaminergic activity and relative abundance of brain derived neurotrophic factor (BDNF) and dopamine receptor mRNA transcripts in the same paradigm. Groups of fish were initially conditioned to associate a flashing light with feeding. Subsequently, the expected food reward was delayed for 30 minutes during two out of three meals per day in the OER treatment, while the previously established routine was maintained in control groups. After 8 days there was no effect of OER on baseline brain stem serotonin (5-HT) or dopamine (DA) activity. Subsequent exposure to acute confinement stress led to increased plasma cortisol and elevated turnover of brain stem DA and 5-HT in all animals. The DA response was potentiated and DA receptor 1 (D1) mRNA abundance was reduced in the OER-exposed fish, indicating a sensitization of the DA system. In addition OER suppressed abundance of BDNF in the telencephalon of non-stressed fish. Regardless of OER treatment, a strong positive correlation between BDNF and D1 mRNA abundance was seen in non-stressed fish. This correlation was disrupted by acute stress, and replaced by a negative correlation between BDNF abundance and plasma cortisol concentration. These observations indicate a conserved link between DA, neurotrophin regulation, and corticosteroid-signaling pathways. The results also emphasize how fish models can be important tools in the study of neural plasticity and responsiveness to environmental unpredictability.

Innovative behaviour in fish: Atlantic cod can learn to use an external tag to manipulate a self-feeder.

This study describes how three individual fish, Atlantic cod (Gadus morhua L.), developed a novel behaviour and learnt to use a dorsally attached external tag to activate a self-feeder. This behaviour was repeated up to several hundred times, and over time these fish fine-tuned the behaviour and made a series of goal-directed coordinated movements needed to attach the feeder's pull string to the tag and stretch the string until the feeder was activated. These observations demonstrate a capacity in cod to develop a novel behaviour utilizing an attached tag as a tool to achieve a goal. This may be seen as one of the very few observed examples of innovation and tool use in fish.

Omission of expected reward agitates Atlantic salmon (Salmo salar).

The evolutionary background for cognition and awareness is currently under ardent scrutiny. Poikilothermic vertebrates such as teleost fishes are capable of classical conditioning and have long-term memories, but it remains unknown to what degree such capabilities are associated with affective states. Here, we investigate whether the concept of frustration may apply to Atlantic salmon. In mammals, this paradigm comprises the omission of an expected reward (OER), which elicits behavioural and physiological coping responses (e.g. aggression and stress reactions). Six groups with 200 fish in each were conditioned to associate a flashing light (CS) with feeding. Conditioning over 22 days led to a change from aversion to attraction to the CS. Subsequently, 3 groups served as control, and 3 groups were subjected to an OER paradigm for 9 days, in which the expected food reward was delayed for 30 min during two out of three daily meals. Compared to controls, OER groups displayed higher levels of aggression and more heterogeneous growth rates, indicating a more pronounced social hierarchy. Cortisol levels did, however, not differ between treatments and both groups responded similarly to acute stress. These results indicate that teleost fishes, like mammals, respond aggressively to OER. The capacity to respond behaviourally to frustrating conditions thus likely reflects an adaptive response to environmental unpredictability, which has been conserved throughout vertebrate evolution.

Food anticipatory behaviour as an indicator of stress response and recovery in Atlantic salmon post-smolt after exposure to acute temperature fluctuation.

In this study we evaluated Pavlovian conditioned food anticipatory behaviour as a potential indicator for stress in groups of Atlantic salmon, and compared this with the physiological stress responses of cortisol excretion into water and hyper-consumption of oxygen. We hypothesised that environmental stress would result in reduced feeding motivation. To assess this, we measured the strength of anticipatory behaviour during a period of flashing light that signalled arrival of food. Further, we expected that fish given a reduced food ration would be less sensitive to environmental stress than fish fed full ration. The fish responded to an acute temperature fluctuation with hyper-consumption of oxygen that decreased in line with the temperature, and elevated cortisol excretion up to 1h after the stressor. These physiological responses did not differ significantly between the food ration groups. The anticipatory behaviour was significantly reduced after the stressor and returned to control levels after 1 to 2 h in the reduced ration group, but not until after 3 to 4 h in the full ration group. Our results show that acute stress can be measured in terms of changes to feeding motivation, and that it is a more sensitive indicator of stress that influences the fish over a longer time period than measures of change in cortisol excretion.

Duration of effects of acute environmental changes on food anticipatory behaviour, feed intake, oxygen consumption, and cortisol release in Atlantic salmon parr.

We compared behavioural and physiological responses and recovery time after different acute environmental challenges in groups of salmon parr. The fish were prior to the study conditioned to a flashing light signalling arrival of food 30 s later to study if the strength of Pavlovian conditioned food anticipatory behaviour can be used to assess how salmon parr cope with various challenges. The effect on anticipatory behaviour was compared to the effect on feed intake and physiological responses of oxygen hyper-consumption and cortisol excretion. The challenges were temperature fluctuation (6.5C° over 4 h), hyperoxia (up to 380% O(2) saturation over 4 h), and intense chasing for 10 min. Cortisol excretion was only elevated after hyperoxia and chasing, and returned to baseline levels after around 3 h or less. Oxygen hyper-consumption persisted for even shorter periods. Feed intake was reduced the first feeding after all challenges and recovered within 3 h after temperature and hyperoxia, but was reduced for days after chasing. Food anticipatory behaviour was reduced for a longer period than feed intake after hyperoxia and was low at least 6 h after chasing. Our findings suggest that a recovery of challenged Atlantic salmon parr to baseline levels of cortisol excretion and oxygen consumption does not mean full recovery of all psychological and physiological effects of environmental challenges, and emphasise the need for measuring several factors including behavioural parameters when assessing fish welfare.

Learning and anticipatory behaviour in a "sit-and-wait" predator: the Atlantic halibut.

We studied the learning capacities and anticipatory behaviour in a "sit-and-wait" predatory fish, the Atlantic halibut, Hippoglossus hippoglossus. In Experiment 1 two groups of halibut received series of light flashes (conditioned stimulus, CS) that started before delivery of food (unconditioned stimulus, US) and persisted until after food delivery, i.e. delay conditioning. Control groups received unpaired CS and US presentations. The anticipatory behaviour of delay conditioned halibut consisted mainly of take-offs towards the surface shortly after onset of the CS. In Experiment 2 six groups of halibut were trained in three trace conditioning procedures: Two groups with 20s, two groups with 60s and two groups with 120s trace interval. Learning was evident in the 20 and 60s trace groups and in one of the 120s trace groups. In contrast to delay conditioning the anticipatory behaviour of trace conditioned halibut was characterized by subtle movements near the tank floor with orientation towards the CS. The cautious responses of halibut after trace conditioning differed markedly from what is observed in other fish species and are suggested to reflect a "sit-and-wait" foraging strategy that requires the predator to remain undetected until the prey is within lunging range.

Sign- and goal-tracking in Atlantic cod (Gadus morhua).

When animals associate a stimulus with food, they may either direct their response towards the stimulus (sign-tracking) or towards the food (goal-tracking). The direction of the conditioned response of cod was investigated to elucidate how cod read cue signals. Groups of cod were conditioned to associate a blinking light (conditioned stimulus, CS) with a food reward (unconditioned stimulus, US), with the CS and the US located at opposite sides of the tank. Two groups were trained in a delay conditioning procedure (CS = 60 s, interstimulus interval = 30 s) and two groups were trained in a trace conditioning procedure (CS = 12 s, trace interval = 20 s). The response pattern was similar for the delay- and trace-conditioned groups. The initial main response at the onset of the CS was approaching the blinking lights, i.e. sign-tracking. In the early trials, the fish did not gather in the feeding area before the arrival of food. In the later trials, the fish first approached the blinking lights, but then moved across the tank and gathered below the feeder before the food arrived, i.e. sign-tracking followed by goal-tracking within each trial. These two responses are interpreted as reflecting two learning systems, i.e. one rapid, reflexive response directed at the signal (sign-tracking) and one slower, more flexible response based on expectations about time and place for arrival of the food (goal-tracking). The ecological significance of these two learning systems in cod is discussed.

Learning in cod (Gadus morhua): long trace interval retention.

Basic knowledge about learning capacities and awareness in fish is lacking. In this study we investigated which temporal gaps Atlantic cod could tolerate between two associated events, using an appetitive trace-conditioning paradigm with blinking light as conditioned stimulus (CS) and dry fish food as unconditioned stimulus (US). CS-US presentations were either temporally overlapping (delay conditioning, CS duration 24 s, interstimulus interval 12 s) or separated by 20, 60, or 120 s (trace conditioning, CS duration 12 s) or 2 h (control, CS duration 12 s). The percentage of fish in the feeding area increased strongly during CS presentation in all delay, 20 s, and 60 s trace groups and in one out of two 120 s trace groups, but not in the control groups. In the 20 and 60 s trace procedures, the fish crowded together in the small feeding area during the trace interval, showing strong anticipatory behaviour. In all the conditioned groups, the fish responded to the CS within eight trials, demonstrating rapid learning. At 88 and 70 days after the end of the conditioning experiments, the delay and 20 s trace groups, respectively, were presented the CS six times at 2-h intervals without reward. All groups responded to the light signal, demonstrating memory retention after at least 3 months. This study demonstrates that Atlantic cod has an impressively good ability to associate two time-separated events and long time retention of learnt associations.