Changes in arterial PO₂, physiological blood parameters and intracellular antioxidants in free-swimming Atlantic cod (Gadus morhua) exposed to varying levels of hyperoxia

Free-swimming Atlantic cod (Gadus morhua) were exposed to water oxygen pressures (P _wO₂) ranging from 18.1 to 41.5 kPa and sampled for blood using an indwelling caudal artery cannula. Arterial blood oxygen pressure (P _aO₂) increased with increasing P _wO₂, from 12.0 kPa in normoxia (18.1 kPa) to 34.2 kPa in the highest hyperoxic level tested (41.5 kPa). Blood CO₂ pressure and plasma bicarbonate concentration increased with P _wO₂, indicating reduced ventilation with increased P _wO₂. Plasma glucose, sodium and potassium were not affected by water oxygen level. Blood oxidative stress biomarkers, reduced glutathione, oxidized glutathione and the oxidative stress index (ratio between oxidized and total glutathione) differed intermittently between normoxia and hyperoxia. The oxidative stress index was higher in the blood of exposed compared to unexposed control cod. Together with elevated P _aO₂, these findings suggest increased production of reactive oxygen species and increased oxidative stress in Atlantic cod exposed to hyperoxia.     

Anaesthesia of Atlantic halibut (Hippoglossus hippoglossus) Effect of pre‐anaesthetic sedation,and importance of body weight and water temperature

The efficacy of the anaesthetic agents benzocaine, metacaine (MS‐222), metomidate, 2‐phenoxyethanol, quinaldine and isoeugenol was studied in Atlantic halibut (Hippoglossus hippoglossus). Fish with an average body weight of 33 g were anaesthetized at 8 °C and fish with an average body weight of 1243 g were anaesthetized at 8 and 15 °C. Agents were tested individually and as combination anaesthesia comprising pre‐anaesthetic sedation, followed by anaesthesia. Induction and recovery times varied in relation to the body weight and water temperature. Large fish had longer induction times and shorter recovery times, and displayed reduced responsiveness to handling compared with small fish. A higher temperature resulted in shorter induction times, longer recovery times and increased responsiveness to handling. Lower dosages were used for all agents in combination anaesthesia. In small fish, this had no effect on the induction times but resulted in shorter recovery times and reduced responsiveness to handling. In large fish, combination anaesthesia resulted in shorter induction times whereas no uniform trend in recovery times and no differences in responsiveness to handling were observed. Neither individual agents nor combinations blocked all reflex reactions to external stimulation in all fish of any treatment group. MS‐222 and benzocaine, used separately or in combination anaesthesia, were the most effective agents in reducing reflex reactions.     

Growth performance,nutrient digestibility and intestinal morphology of rainbow trout (Oncorhynchus mykiss) fed graded levels of the yeasts Saccharomyces cerevisiae and Wickerhamomyces anomalus

In a 10‐week study, we evaluated the effects of replacing 20%, 40% or 60% of fishmeal (present in control diet at 300 g/kg) on a digestible protein basis with yeast Saccharomyces cerevisiae or a yeast mixture of Wickerhamomyces anomalus and S. cerevisiae on growth performance, nutrient digestibility, nutrient retention and intestinal health of rainbow trout (Oncorhynchus mykiss). Triplicate tanks with 35 rainbow trout (144.7 ± 25.1 g mean ± SEM) were fed rations of 1.5% of total biomass per tank. Replacement of 60% of fishmeal with yeast mixture resulted in lower specific growth rate of 1.0% versus 1.2%/day for other diets. Apparent digestibility coefficients for crude protein and most amino acids were highest in fish fed fishmeal‐based diet, with similar values for fish fed the diet with 20% replacement with yeast mixture. Diet with 20% replacement with yeast mixture resulted in highest phosphorus digestibility. Replacement of 60% of fishmeal with S. cerevisiae resulted in oedematous mucosal fold tips in the proximal intestine. The results of this study suggest that these yeasts can replace up to 40% of fishmeal under current inclusion levels in diets for rainbow trout without compromising growth performance, nutrient digestibility or intestinal health.     

A test of the individual feeding activity and food size preference in rainbow trout using demand feeders

Feeding activity and food size preferences were investigated in three groups of rainbow trout with initial weights ranging from 200 to 1000 g. Fish were given the option of releasing food from either of two demand feeding devices that contained either 6 mm or 9 mm food pellets. Individual biting activity (to activate release of food) was continuously monitored using specially designed passive integrated transponders (PIT-tags). In each of the groups, biting activity was strongly influenced by three individuals, which accounted for 70–85% of the total. Thus, high bite activity by few individuals indicates the development of social dominance hierarchies. There was no statistically significant correlation between number of bites and growth rate, indicating that the fish that operated the demand feeders did not gain any growth advantage. Trout that dominated the actuation of the feeders were found among both medium- and large-sized fish. There was no evidence of active food size selection, and the fish that most frequently operated the feeders defended one area of the tank rather than preferring the trigger that would have delivered food of the optimal size.     

Genetic differentiation between and within ecotypes of pike (Esox lucius) in the Baltic Sea

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Temperature‐dependent feed requirements in farmed blue mussels (Mytilus edulis L.) estimated from soft tissue growth and oxygen consumption and ammonia‐N excretion

Feed requirements were estimated from specific growth rates in standardized soft tissue dry weight (SGR_DW’) and atomic O:N ratios for mussels fed seven rations of microalgae (5–735 μg C h^?1 ind^?1) at 7 and 14°C respectively. The mean oxygen consumption and ammonia‐N excretion rates were significantly higher at 14°C (0.29 μg O² and 27.3 μg N ind^?1 h¹) compared with those at 7°C (0.16 μg O² and 11.4 μg N ind^?1 h^?1) (P < 0.05), resulting in O:N ratios between 3 and 45 at 7°C and 7 and 28 at 14°C. Low O:N ratios indicate protein catabolism and an unfavourable condition, whereas high ratios indicate that carbohydrate is the primary energy source. The measured SGR_DW’ suggests minimum feed requirements of ~240 and ~570 μg C ind^?1 h^?1 for weight maintenance at 7 and 14°C, with corresponding O:N ratios of 24 and 16, respectively, indicating a more stressed condition at 14°C. A 0.5% SGR_DW’ day^?1 was obtained by ~565 (O:N = 29) and ~680 (O:N = 23) μg C ind^?1 h^?1 at 7 and 14°C respectively. A positive and significantly higher SGR_DW’, with the lowest feed ration at 7°C compared with a negative SGR_DW’ at 14°C (P < 0.05), indicated that storage time can also possibly be prolonged at low temperatures if the mussels are not fed.     

Prediction of Starch,Soluble Sugars and Amino Acids in Potatoes (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Using Hyperspectral Imaging,Dielectric and LF-NMR Methodologies

Handling and processing of potatoes is performed in increasingly large and more automated facilities, and the industry calls for more automated machinery for quality assessment and sorting by concentration of starch, soluble sugars, protein, amino acids etc. of the potato tubers. The present study was designed to evaluate five different scanning methods for their potential use in potato assessment and sorting. Two methods were based on hyperspectral imaging, two were based on dielectric/bio-impedance and one was based on low-field nuclear magnetic resonance. A set of 60 potatoes of 10 different cultivars were simultaneously sampled for analyses of content and scanned by the five different scanning methods. The resulting multivariate dataset was used to estimate the prediction ability of the individual scanning methods on starch-related parameters, selected simple sugars, selected amino acids, conductivity of pressed cell sap and cell sizes. Results showed that most types of spectral analyses had relatively high potential for predicting the starch-related parameters and medium potential for predicting the concentration of the reducing sugars fructose and glucose. Most methods showed medium potential for prediction of several amino acids, including asparagine, which showed particularly promising predictions in the hyperspectral analyses of intact potatoes. The presented screening study enabled us to perform robust choices for the further development and optimization of the methods and instruments for industrial implementation.     

Automated image analysis as a tool to quantify the colour and composition of rainbow trout (Oncorhynchus mykiss W.) cutlets

The goal of this paper is to propose and evaluate automated image analysis methods for describing muscle cutlets in rainbow trout. The proposed automated image analysis methods were tested on a total of 983 scanned images of trout cutlets, and included quality traits such as fat percentage, flesh colour and the size of morphologically distinguishable subparts of the cutlet. A sub-sample of 50 images was randomly selected for manual segmentation of the cutlet, the dorsal fat depot and the red muscle and regions. The identification of these regions by manual and automatic image analysis correlated strongly (r = 0.97, r = 0.95 and r = 0.91, respectively). The estimated fat percentage obtained from image analysis, based on the area of visible fat and the colour of the cutlet flesh, correlated well with chemical fat percentage measured by mid-infrared transmission spectroscopy (MIT) (r = 0.78). The automated image analysis methods are therefore a reliable means of predicting the fat percentage of trout cutlets. Principal component analysis (PCA) loading plots were used to identify subsets of variables from the image analysis of special significance for further studies; cutlet area, dorsal fat depot area, red muscle area, back height, cutlet width, and width of left and right abdomen wall were among the variables selected. PCA loading plots of different colour variables indicated that simple statistical coefficients such as percentiles and mean values can be used to quantify different aspects of flesh colour. In conclusion, the methods presented here provide a powerful toolbox for describing important morphological structures and quality traits of trout cutlets.     

Surplus spermine does not protect salmon cells against lipopolysaccharide (LPS)‐induced stress,but excess spermine is eliminated through spermidine/spermine N1‐acetyltransferase (SSAT)

The present trials tested the efficiency of surplus spermine to reduce inflammation and oxidative stress following LPS‐induced stress using an in vitro model of head kidney and liver cells isolated from Atlantic salmon. Spermine did not protect cells from LPS‐induced inflammatory response at either 0.3, 0.6 or 0.9 mM. However, as the gene expression of spermidine/spermine N1‐acetyltransferase (SSAT) increased with increasing spermine concentration, we addressed possible oxidative effects of the increased SSAT using its activator DENSPM or inhibitor of polyamine oxidation of the acetylated polyamines using MDL72527 at a spermine concentration of 0.6 mM. There was no significant effect of DENSPM, but MDL72527 decreased gene expression of GPX‐3 (p = .04), while gene expression of catalase and MnSOD was unaffected by treatment (p = .30 and p = .48, respectively). In conclusion, spermine did not protect cells from LPS‐provoked inflammation. The higher the spermine concentration, the more SSAT producing acetylated spermine occurred. Inhibiting the acetylated polyamine oxidases by MDL72527 improved oxidation status as expected due to a lower endogenous production of H₂O₂ by polyamine and acetylated polyamine oxidases. Probably care should be taken using polyamines or arginine as functional ingredients to avoid any increased oxidation within cells.     

Anaesthesia of farmed fish: implications for welfare

During their life cycle as farmed animals, there are several situations in which fish are subjected to handling and confinement. Netting, weighing, sorting, vaccination, transport and, at the end, slaughter are frequent events under farming conditions. As research subjects, fish may also undergo surgical procedures that range from tagging, sampling and small incisions to invasive procedures. In these situations, treatment with anaesthetic agents may be necessary in order to ensure the welfare of the fish. The main objective of this paper is to review our knowledge of the effects of anaesthetic agents in farmed fish and their possible implications for welfare. As wide variations in response to anaesthesia have been observed both between and within species, special attention has been paid to the importance of secondary factors such as body weight, water temperature and acute stress. In this review, we have limited ourselves to the anaesthetic agents such as benzocaine, metacaine (MS-222), metomidate hydrochloride, isoeugenol, 2-phenoxyethanol and quinaldine. Anaesthetic protocols of fish usually refer to one single agent, whereas protocols of human and veterinary medicine cover combinations of several drugs, each contributing to the effects needed in the anaesthesia. As stress prior to anaesthesia may result in abnormal reactions, pre-anaesthetic sedation is regularly used in order to reduce or avoid stress and is an integral part of the veterinary protocols of higher vertebrates. Furthermore, the anaesthetic agents that are used in order to obtain general anaesthesia are combined with analgesic agents that target nociception. The increased use of such combinations in fish is therefore included as a special section. Anaesthetic agents are widely used to avoid stress during various farming procedures. While several studies report that anaesthetics are effective in reducing the stress associated with confinement and handling, there are indications that anaesthesia may in itself induce a stress response, measured by elevated levels of cortisol. MS-222 has been reported to elicit high cortisol release rates immediately following exposure, while benzocaine causes a bimodal response. Metomidate has an inhibitory effect on cortisol in fish and seems to induce the lowest release of cortisol of the agents reported in the literature. Compared to what is observed following severe stressors such as handling and confinement, the amount of cortisol released in response to anaesthesia appears to be low but may represent an extra load under otherwise stressful circumstances. Furthermore, anaesthetics may cause secondary adverse reactions such as acidosis and osmotic stress due to respiratory arrest and insufficient exchange of gas and ions between the blood and the water. All in all, anaesthetics may reduce stress and thereby improve welfare but can also have unwanted side effects that reduce the welfare of the fish and should therefore always be used with caution. Finally, on the basis of the data reported in the literature and our own experience, we recommend that anaesthetic protocols should always be tested on a few fish under prevailing conditions in order to ensure an adequate depth of anaesthesia. This recommendation applies whether a single agent or a combination of agents is used, although it appears that protocols comprising combinations of agents provide wider safety margins. The analgesic effects of currently used agents, in spite of their proven local effects, are currently being debated as the agents are administrated to fish via inhalation rather than locally at the target site. We therefore recommend that all protocols of procedures requiring general anaesthesia should be complemented by administration of agents with analgesic effect at the site of tissue trauma.