Proof of concept: visual categorization of carotenoid pigmentation in Arctic charr (Salvelinus alpinus L) can predict stress response.

Carotenoid pigmentation in Salvelinus alpinus has been connected to stress responsiveness in earlier studies. This has, however, only been tested with time-consuming image analysis from photos. Here, we used quick visual categorization of carotenoid pigmentation to investigate the stress responsiveness of the extreme groups. The visually selected charr were then exposed to a net restraint stressor. Arctic charr with few spots also had a lower stress responsiveness compared to charr with many spots. Thus, visual selection could be used as a simple method within aquaculture.

Metabolomics Approach To Evaluate a Baltic Sea Sourced Diet for Cultured Arctic Char (Salvelinus alpinus L.).

Aqua feeds traditionally rely on fishmeal as a protein source, which is costly and unsustainable. A new feed was formulated in the study with Baltic Sea sourced decontaminated fishmeal, Mytilus edulis and Saccharomyces cerevisiae, and given to Arctic char (Salvelinus alpinus) for ten months. The diet-induced changes on metabolic profile in fish plasma, liver, and muscle were studied relative to a fishmeal-based standard diet by using a 1H NMR-based metabolomics approach. Fish fed the test diet had higher content of betaine and lower levels of trimethylamine-N-oxide and aromatic amino acids in plasma or tissues, which were mainly caused by the diet. The metabolomics results are useful to understand the mechanism of lower body mass, smaller Fulton's condition factor, and a tendency of less lipid content observed in fish fed the test diet. Thus, modifications on the dietary levels of these compounds in the feed are needed to achieve better growth performance.

Brain cortisol receptor expression differs in Arctic charr displaying opposite coping styles.

Individually consistent behavioral and physiological responses to stressful situations (often referred to as coping styles) has been reported in many animal species. Differences in hypothalamic-pituitary axis reactivity characterize individuals, and it has been proposed that the glucocorticoid (gr) and mineralocorticoid (mr) receptors are fundamental in regulating coping styles. We sorted individuals into reactive and proactive coping styles by collapsing behavioral outputs from net restraint and confinement stress tests in a principal component analysis. We then analyzed plasma cortisol levels, serotonin neurochemistry and the relative mRNA expression of gr1 and mr in stressed individuals per coping style. Proactive fish were characterized as having a lower serotonergic activity and being more active under stress. In addition, proactive fish had higher hypothalamic gr1 and mr abundance and a higher mr/gr1 ratio, compared to reactive fish. We found no significant differences in cortisol or telencephalic mRNA, gr1 and mr expression, or their ratio. Brain MR and GR have been proven to have an important role in the appraisal, coping and adaptation to stressful stimuli, so that a higher expression of these receptors in proactive fish suggests increased tolerance and performance under stress, compared to reactive individuals. We present evidence of a conserved neuroendocrine mechanism associated with coping styles in a fish species which is ecologically very diverse and considered to be the most cold-adapted fish in freshwater. We propose that this may be a first step into exploiting this model in order to better understand climate-change related effects in sub populations and ecophenotypes.

Social stress effects on pigmentation and monoamines in Arctic charr.

Pigmentation often signals status and in general melanin-based pigmentation is indicative of aggression and stress resilience in vertebrates. This is evident in the salmonids Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) where more melanin spotted individuals are more stress resilient. However, in the salmonid Arctic charr (Salvelinus alpinus) it seems as if it is carotenoid-based pigmentation that signals aggression and stress resilience. In our study, social stress effects on carotenoid-based spots, and behavioural and physiological stress responses were investigated. Socially stressed individuals have more spots, and behavioural stress responses were associated with spots. Some of the results concerning physiological stress responses, such as plasma cortisol levels and monoaminergic activity, are associated with spottiness. Further, the earlier proposed lateralization of spots, with left side connected to stress responsiveness and right side to aggression, is to some extent validated although not conclusively. In conclusion, this study provides further evidence that more stressed charr have more carotenoid spots, and for the first time monoaminergic activity is shown to be connected with carotenoid pigmentation.

Dominance and stress signalling of carotenoid pigmentation in Arctic charr (Salvelinus alpinus): lateralization effects?

Social conflicts are usually solved by agonistic interactions where animals use cues to signal dominance or subordinance. Pigmentation change is a common cue used for signalling. In our study, the involvement of carotenoid-based pigmentation in signalling was investigated in juvenile Arctic charr (Salvelinus alpinus). Size-matched pairs were analysed for pigmentation both before and after being tested for competitive ability. We found that dominant individuals had fewer carotenoid-based spots on the right and left sides as well as lower plasma cortisol levels compared to subordinate individuals. Further, the number of spots on both sides was positively associated with plasma cortisol levels. These results indicate that carotenoid-based pigmentation in Arctic charr signals dominance and stress coping style. Further, it also appears as if carotenoid-based pigmentation is lateralized in Arctic charr, and that the right side signals aggression and dominance whereas the left side signals stress responsiveness.

Toxicity and accumulation of trinitrotoluene (TNT) and its metabolites in Atlantic salmon alevins exposed to an industrially polluted water.

A pond in an industrial area in Sweden was selected to study adverse effects on salmon alevins from 2,4,6-trinitrotoluene (TNT)-contaminated water. Chemical screening revealed heavy contamination of TNT and its degradation products, 2-amino-4,6-dinitrotoluene (2-ADNT) and 4-amino-2,6-dinitrotoluene (4-ADNT), ranging from 0.05 to 230 g/kg in the sediment (dry weight) within the water system. Pond water contained 3 mg/L TNT. A dilution series of pond water mixed with tap water revealed increased death frequency in alevins down to fivefold dilution (approximate 0.4 mg TNT/L). Uptake was concentration dependent, reaching 7, 9, and 22 µg/g tissue for TNT, 2-ADNT, and 4-ADNT at the highest test concentration. A time-dependent uptake of TNT and its degradation products was found at a water concentration of 0.08 mg TNT/L. Degradation products of TNT showed a more efficient uptake compared to native TNT, and accumulation of 4-ADNT was more pronounced during the late phase of the 40-d exposure study. Bioconcentration factors (BCF) (0.34, 52, and 134 ml/g for TNT, 2-ADNT, and 4-ADNT, respectively) demonstrated a significant uptake of the metabolite 4-ADNT in alevin tissue. Disturbed physiological conditions and delayed development in alevins were not studied, but may not be excluded even at 125-fold diluted pond water (0.016 mg TNT/L). BCF data indicated that bioaccumulation of TNT metabolites need to be considered in TNT chronic toxicity. Fish species and age differences in the accumulation of TNT metabolites need to be further studied.

Evaluation of chitinolytic activities and membrane integrity in gut tissues of Arctic charr (Salvelinus alpinus) fed fish meal and zygomycete biomass.

Chitinolytic activity, nutrient uptake and intestinal barrier functions were investigated in gut tissues of Arctic charr (Salvelinus alpinus) fed iso-nitrogenous diets based on fish meal, with or without inclusion of zygomycetes (Rhizopus oryzae). We found that gut tissue of Arctic charr had significant chitinase activity, of both endo- and exo-chitinase iso-forms. Moreover, the distribution pattern along the GI tract of Arctic charr differed between endo-chitinase and exo-chitinase. The endo-chitinase activity in stomach tissue and in the distal intestine was several hundred-fold higher than the exo-chitinase activity in stomach tissue. The greatest exo-chitinase activity was found in the distal intestine. The zygomycete-based diet resulted in higher chitinolytic activity in gut tissue compared to the fish meal-based diet. Disturbed intestinal integrity and increased uptake rate of the amino acid lysine were observed in the distal, but not proximal, intestine of fish fed the zygomycete-based feed.

High-resolution ¹H magic angle spinning NMR spectroscopy of intact Arctic char (Salvelinus Alpinus) muscle. Quantitative analysis of n-3 fatty acids, EPA and DHA.

The lipid and small metabolite profiles from intact muscles of Arctic char were investigated using (1)H high-resolution magic angle spinning ((1)H HR-MAS) NMR spectroscopy. Not only the total n-3 fatty acid content but also the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) contents of the muscle were obtained from the (1)H HR-MAS NMR spectra without pretreatment of the tissue or lipophilic extraction. A number of small metabolites could also be observed, where creatine/phosphocreatine, anserine and taurine were the most abundant. Thus, the use of (1)H HR-MAS NMR led to simplified analysis techniques that can give direct information on the nutritional value of the fish.

Potential toxic effect on aquatic fauna by the dwarf shrub Empetrum hermaphroditum.

The common evergreen dwarf shrub Empetrum hermaphroditum has influence on the functioning of boreal terrestrial ecosystems in northern Sweden. The negative effects of E. hermaphroditum are partly attributed to the production of the dihydrostilbene, batatasin-III, which is released from leaves and litter by rain and snowmelt. In this study, we investigated whether batatasin-III is carried by runoff into streams and lakes during the snowmelt period and whether it is also potentially hazardous to aquatic fauna. Sampling of water from streams and a lake for which the surrounding terrestrial vegetation is dominated by E. hermaphroditum was done during the snowmelt period in May 1993 and in 1998, and analyzed for batatasin-III. Using 24- and 48-hr standard toxicity tests, we analyzed toxicity to brown trout (Salmo trutta) alevins and juvenile water fleas (Daphnia magna). Toxicity (proportion of dead individuals) to trout was tested at pH 6.5 and compared with that of a phenol within a range of concentrations. In the toxicity (proportion of immobilized individuals) test on D. magna, the interactive effect of pH (pH 5.5-7.0) was included. Concentration of batatasin-III was generally higher in 1998 than in 1993 and showed peak levels during snowmelt. Concentration in ephemeral runnels > the lake > streams running through clear-cuts dominated by E. hermaphroditum > control streams lacking adjacent E. hermaphroditum vegetation. The maximum concentration of batatasin-III found was 1.06 mg l(-1). The proportion of dead yolk sac alevins increased significantly (P < 0.001) with increasing concentrations of batatasin-III and time of exposure. After 24 hr, EC50 was 10 mg l(-1). It was 2 mg l(-1) after 48 hr. The effect of phenol was negligible, indicating a specific phytotoxic effect of the bibenzyl structure of batatasin-III. The proportion of mobile D. magna became significantly smaller (P < 0.001) with increasing concentrations of batatasin-III, with decreasing pH, and with increasing exposure time. EC50 varied between 7 and 17 mg l(-1) at pH 5.5 and 7.0, respectively. After 24 hr EC50 decreased and was 2.5 at pH 5.5 and 12 mg l(-1) at pH 7.0. The levels of batatasin-III found in the field samples were below the lowest EC50 in acute toxicity tests. However, in view of the interactive effect of pH and exposure time, this study suggests that this stable plant metabolite may impose a lethal effect on the aquatic fauna in small streams.