Lars Helge Stien

The interaction between water currents and salmon swimming behaviour in sea cages.

Positioning of sea cages at sites with high water current velocities expose the fish to a largely unknown environmental challenge. In this study we observed the swimming behaviour of Atlantic salmon (Salmo salar L.) at a commercial farm with tidal currents altering between low, moderate and high velocities. At high current velocities the salmon switched from the traditional circular polarized group structure, seen at low and moderate current velocities, to a group structure where all fish kept stations at fixed positions swimming against the current. This type of group behaviour has not been described in sea cages previously. The structural changes could be explained by a preferred swimming speed of salmon spatially restricted in a cage in combination with a behavioural plasticity of the fish.

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.

Soft texture of atlantic salmon fillets is associated with glycogen accumulation.

Atlantic salmon (Salmo salar L.) with soft fillets are not suited for manufacturing high quality products. Therefore fillets with insufficient firmness are downgraded, leading to severe economic losses to the farming and processing industries. In the current study, morphological characteristics of salmon fillets ranging from soft to hard were analysed. Different microscopic techniques were applied, including novel methods in this field of research: morphometric image analysis, periodic acid Schiff staining, immunofluorescence microscopy, transmission electron microscopy and fourier transform infrared microscopy. The results showed that the myocytes of soft muscle had detached cells with mitochondrial dysfunctions, large glycogen aggregates and enlarged inter cellular areas, void of extracellular matrix proteins, including lower amounts of sulfated glycoproteins. Myofibre-myofibre detachment and disappearance of the endomysium in soft muscles coincided with deterioration of important connective tissue constituents such as Collagen type I (Col I), Perlecan and Aggrecan. In summary our investigations show for the first time an association between soft flesh of Atlantic salmon and massive intracellular glycogen accumulation coinciding with degenerated mitochondria, myocyte detachment and altered extracellular matrix protein distribution. The results are important for further understanding the etiology of soft salmon.

Predicting the effectiveness of depth-based technologies to prevent salmon lice infection using a dispersal model.

Salmon lice is one of the major parasitic problems affecting wild and farmed salmonid species. The planktonic larval stages of these marine parasites can survive for extended periods without a host and are transported long distances by water masses. Salmon lice larvae have limited swimming capacity, but can influence their horizontal transport by vertical positioning. Here, we adapted a coupled biological-physical model to calculate the distribution of farm-produced salmon lice (Lepeophtheirus salmonis) during winter in the southwest coast of Norway. We tested 4 model simulations to see which best represented empirical data from two sources: (1) observed lice infection levels reported by farms; and (2) experimental data from a vertical exposure experiment where fish were forced to swim at different depths with a lice-barrier technology. Model simulations tested were different development time to the infective stage (35 or 50°-days), with or without the presence of temperature-controlled vertical behaviour of lice early planktonic stages (naupliar stages). The best model fit occurred with a 35°-day development time to the infective stage, and temperature-controlled vertical behaviour. We applied this model to predict the effectiveness of depth-based preventive lice-barrier technologies. Both simulated and experimental data revealed that hindering fish from swimming close to the surface efficiently reduced lice infection. Moreover, while our model simulation predicted that this preventive technology is widely applicable, its effectiveness will depend on environmental conditions. Low salinity surface waters reduce the effectiveness of this technology because salmon lice avoid these conditions, and can encounter the fish as they sink deeper in the water column. Correctly parameterized and validated salmon lice dispersal models can predict the impact of preventive approaches to control this parasite and become an essential tool in lice management strategies.

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.

Different Dietary Levels of Protein to Lipid Ratio Affected Digestive Efficiency, Skeletal Growth, and Muscle Protein in Rainbow Trout Families

Normal Protein (NP) and High Protein (HP) diets were provided to rainbow trout families from juvenile to maturity. Muscle protein concentration increased during growth and the protein to lipid (P/L) ratio was doubled at late stage with respect to the dietary P/L ratio. The HP-diet fish showed higher protein deposition in body and white muscle, and had lower condition factor due to protein deposition associated more with body length than body weight. Fish growth rates were decreased at maturation, and the HP-diet fish showed lower growth rate including the activity ratio of trypsin to chymotrypsin (T/C ratio) and feed efficiency. Trypsin and chymotrypsin specific activities were related to dietary protein levels, and the T/C ratio was related to intestinal weight and growth rate independent of the enzymes specific activity levels. The families with high growth capacity could double increase white muscle P/L ratio levels, compared to low and medium growth families, if they were fed on HP-diet. The digestive enzyme extracts from high growth families resulted in higher in vitro protein digestibility for all diets. The effect of dietary P/L ratio on digestion, skeletal growth (length) and the white muscle P/L ratio in fish with different growth capacities is illustrated.

Lowering treatment temperature reduces salmon mortality: a new way to treat with hydrogen peroxide in aquaculture

BACKGROUND Hydrogen peroxide (H2 O2 ) baths are widely used to reduce numbers of salmon lice on farmed Atlantic salmon. Fish mortalities often occur after baths, with warmer temperatures increasing lethality. We tested whether mortality could be reduced and lice removal efficacy maintained by lowering bath temperatures relative to ambient temperatures. Post-smolt salmon infected with lice were held at 10, 13 or 16 °C, and treated with 1.5 g/L H2 O2 for 20 min at equal or lower bath temperatures of 7, 10 or 13 °C. RESULTS Salmon mortality decreased as ambient and bath temperatures decreased. No mortality occurred when fish at 13 °C were treated at 7 °C. For ambient temperatures of 16 °C, the number of lice remaining was reduced by four-fold when treated at 7 °C compared with 13 °C. All treatments in which mortality was zero had similar efficacies regardless of bath temperature. CONCLUSION We took salmon from warmer to colder temperatures to determine the optimum bathing temperature to prevent mortality. A temperature of 7 °C was optimal when treating with 1.5 g/L of H2 O2 , as mortality was zero and pre-adult lice removal was unchanged. By manipulating temperature, we developed a new method of H2 O2 bathing that reduces mortality. When ambient temperatures are >10 °C, we recommend that the industry decrease H2 O2 bath temperatures.

Consistent melanophore spot patterns allow long‐term individual recognition of Atlantic salmon Salmo salar

The present study shows that permanent melanophore spot patterns in Atlantic salmon Salmo salar make it possible to use images of the operculum to keep track of individual fish over extended periods of their life history. Post-smolt S. salar (n = 246) were initially photographed at an average mass of 98 g and again 10 months later after rearing in a sea cage, at an average mass of 3088 g. Spots that were present initially remained and were the most overt (largest) 10 months later, while new and less overt spots had developed. Visual recognition of spot size and position showed that fish with at least four initial spots were relatively easy to identify, while identifying fish with less than four spots could be challenging. An automatic image analysis method was developed and shows potential for fast match processing of large numbers of fish. The current findings promote visual recognition of opercular spots as a welfare-friendly alternative to tagging in experiments involving salmonid fishes.

Effect of Blood Removal Protocol and Superchilling on Quality Parameters of Prerigor Filleted Farmed Atlantic Cod (Gadus morhua)

UNLABELLED A total of 40 farmed Atlantic cod (Gadus morhua) were in 2 groups either fillet directly after stunning and spray washed or produced into fillets according to traditional slaughter methods including exsanguination for 30 min, gutting and washing. Both groups were either stored superchilled or traditionally on ice. After 7 d postmortem color (CIE L*, a*, b*) and fillet shrinkage was measured by computer imaging along with drip loss and texture hardness. Results show that superchilled fillets had significant lower core temperature than fillets stored on ice during the entire 7 d storage period. This resulted in reduced fillet shrinkage from 14.7% to 6.9% and less drip loss dropping from 9.45% to 3.99% in average. Processing the fish directly into fillets resulted in satisfactory blood drainage, where all groups were in particularly well exsanguinated with a* values below zero. No color difference was observed between filleting groups or chilling methods. Spray washing of the fillets resulted in water uptake and higher drip loss in interaction with chilling method. We conclude that filleting farmed fish in one step is feasible. PRACTICAL APPLICATION Traditionally farmed fish are slaughtered and processed over several steps, which often include live chilling, stunning, exsanguination, chilling, gutting, rinsing, decapitation, filleting before the fillets are packed into polystyrene boxes and shipped with ice. These processes are often time, laboring, space, and energy consuming. A novel processing line for filleting of farmed fish is gutting and filleting the fish directly after decapitation and replacing exsanguination with spray washing the fillets. In addition, all the cooling steps are replaced by superchilling the fillets. This novel process line gives fillets with comparable if not superior quality compared to the traditional process.