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The aim of the current study was to investigate how freshwater diets impact on immunity in Atlantic salmon smolts in freshwater, during transfer to seawater and in post smolts during the seawater stage with and without pancreas disease (PD) infection. Three specific freshwater diets were prepared: (i) A diet similar in composition to commercial salmon freshwater diets (Standard diet); (ii) A diet composed of vegetable oils (rapeseed, palm and linseed oils) mimicking the fat composition in aquatic insects – the natural diet of wild salmon in freshwater (Fatty acid diet); (iii) A diet enriched with possible immune modulating amino acids including dl -methionine, l -lysine, l -threonine and taurine (Amino acid diet). After seawater transfer, all fish were fed the same commercial diet. Head kidneys were extracted, and their leukocytes isolated from smolts right before transfer to seawater, from post smolts one and six weeks after transfer to seawater, and from post smolts in seawater after 8 weeks of ongoing PD infection. In addition, to provoke bacterial or virus induced inflammation in vitro, the individual leukocyte suspension from all fish were stimulated by lipopolysaccharide (LPS) or polyinosinic acid: polycytidylic acid (PIC). The transfer of smolts from fresh-to seawater changed the transcription of several types of genes. Particularly in isolates from fish fed the Standard or Fatty acid diet in freshwater, overall gene transcription (IL-1β , CD83 , INF-γ , cox2 , cd36 , MGAT2, catalase) declined. However, the Amino acid diet stimulated the LPS induced gene transcription of IL-1β , CD83 , Cox2 , and INF-γ at this stage. In freshwater smolts, PIC stimulated leukocytes showed higher transcription level of Mx and viperin in the Fatty acid and Amino acid diet groups compared to the Standard diet group. In seawater post smolts, Mx and viperin responded similarly to PIC challenge in all diet groups. Furthermore, leukocytes isolated from PD infected fish, continued responding to PIC, regardless of freshwater diet. • Modified Amino- and Fatty acid composition in freshwater diets can increase the immunological robustness of salmon parr towards virus infections during the freshwater phase. • The Amino acid composition in freshwater diets can be tailored to be protective against bacterial and oxidative damage in post smolts newly transferred to seawater. • When challenged with PIC, PD virus or PD virus + PIC, salmon immune cells use their energy to produce antiviral molecules, regardless of diet composition in freshwater. [ABSTRACT FROM AUTHOR]

Methionine-choline-deficient ( MCD) mammals are known to accumulate liver TAG probably due to phosphatidylcholine ( PC) deficiency and thus assembly of VLDL and transport of lipids from liver to peripheral organs. To assess whether supplementation of choline could spare methionine and secure a healthy liver metabolism, by reducing the endogenous PC synthesis without interfering with lipid transport and distribution, Atlantic salmon with initial BW of 700 g were fed adequate (1.9 g Met/16 gN) or surplus methionine (2.5 g Met/16 gN) diets of which were supplemented with choline or not for a period of 19 weeks. Fish fed the low-methionine diets had reduced growth ( p = .013) due to reduced protein gain ( p = .007), while lipid gain slightly improved in fish fed the choline-supplemented diets ( p = .047). Also, feed conversion improved when fed surplus methionine ( p < .001), while choline supplementation had no impact on feed conversion. No interaction between choline and methionine on growth performance or retention existed. Phospholipid status in liver and muscle was not affected by treatments, and no liver TAG accumulation occurred at the methionine levels used. Gene expression of ApoB100 necessary for assembling VLDL or pemt necessary for endogenous PC synthesis was unaffected by treatments. Capacity of methylation ( MAT, BHMT) within the liver was not affected by treatment nor was the gene expression of enzymes in liver transsulfuration ( CBS or CDO). Methionine status within liver was unaffected by treatments, while free methionine reduced in those fish fed the low-methionine diets in muscle and plasma. Cystathionine and taurine were elevated when fed surplus methionine. Choline supplementation had no impact on sulphur amino acid metabolites in either tissue. Neither did choline supplementation improve TAG mobilization from liver to muscle. To conclude, choline does not improve endogenous phospholipid synthesis or transport of TAG from liver to muscle depot when added to diets containing 1.9 g Met/16 gN, while surplus methionine improved growth and protein retention, indicating that 1.9 g Met/16 gN is enough to support a healthy liver metabolism, but too low to support muscle protein deposition in adult salmon fed high plant protein diets for longer periods of time. [ABSTRACT FROM AUTHOR]

Phosphatidylcholine is synthesized endogenously through the enzyme phosphatidylethanolamine-methyl transferase. As endogenous choline synthesis requires methyl groups from S-adenosylmethionine (SAM), the endogenous synthesis of choline may depend on SAM availability. SAM availability depends on methionine and ATP. SAM is also a precursor for cysteine and may affect taurine and glutathione concentrations. To investigate whether choline synthesis or transsulphuration is prioritized and the interactions between taurine and choline supplementation when methionine availability is low, juvenile Atlantic salmon were fed low methionine diets with and without taurine and choline supplementation. There were no differences in growth or protein accretion following treatments. Fish fed the low methionine diets did not develop a fatty liver, but choline supplementation increased the concentration of total phospholipids in liver and muscle. Taurine supplementation increased taurine concentrations in liver and muscle, but no interactions with choline were present. Liver SAM was unaffected by treatments. Two of three tanks fed the low methionine diet without choline and taurine supplementation had a higher gene expression of p38MAPK (mitogen activated phosphokinase). Choline supplementation to low methionine diets thus may have beneficial effect on the metabolic health and lipid transport to extra-hepatic tissues. Taurine supplementation had no effect on phospholipids status, but increased taurine concentration in tissues and as such may affect oxidation status when methionine availability is low. [ABSTRACT FROM AUTHOR]

The current study aimed to investigate the effect of surplus dietary arginine on polyamine and amino acid metabolism and accretion of proteins and lipids, as previous studies have demonstrated a lipid-reducing effect following surplus arginine supplementation in viscera as well as increased muscle growth. Four plant protein-based diets were given increasing concentrations of L-arginine, from 21.1 to 36.1 g kg-1 DM, and fed to quadruplicate tanks of adult Atlantic salmon (1.1 kg) for 12 weeks. No effects on growth or deposition of fat or protein were observed. Free amino acid concentrations and related metabolites were unaffected in the liver, except for urea, while concentrations in muscle and plasma reflected production of arginine metabolites. Polyamine concentrations were unaffected in liver, muscle and white adipose tissue ( WAT), as were the abundance and activity of spermidine/spermine N1-acetyltransferase ( SSAT), the rate-limiting enzyme in polyamine turnover. Gene expression demonstrated differential regulation of ornithine decarboxylase ( ODC) in liver and WAT, although overall little effects were observed on gene expression. Liver S-adenosylmethionine ( SAM) concentrations decreased with arginine supplementation. We suggest that adult Atlantic salmon have adapted to surplus arginine, and the main long-term effect appears to be increased concentrations of arginine metabolites. [ABSTRACT FROM AUTHOR]

Abstract: Head kidney leukocytes isolated from Atlantic salmon fed either a diet based on fish oil (FO) or soy bean oil (VO) were used in order to evaluate if different lipid sources could contribute to cellular activation of the salmon innate immune system. A specific inhibitor of p38 MAPK, SB202190, was used to investigate the effect of lipopolysaccharide (LPS) signalling in the head kidney leukocytes. The results show that LPS up regulate IL-1β, TNF-α, Cox2 expression in leukocytes isolated from fish fed either diet. The p38 MAPK inhibitor, SB202190, reduced the LPS induced expression of these genes in both dietary groups. In LPS stimulated leukocytes isolated from VO fed fish, SB202190 showed a clear dose dependent inhibitory effect on IL-1β, TNF-α and Cox2 expression. This effect was also observed for Cox2 in leukocytes isolated from FO fed fish. Furthermore, there was a stronger mean induction of Cox2 in LPS stimulated leucocytes isolated from the VO-group compared to LPS stimulated leukocytes isolated from the FO-group. In both dietary groups, LPS stimulation of salmon head kidney leukocytes increased the induction of CD83, a dendrite cell marker, while the inhibitor reduced CD83 expression in the VO fed fish only. The inhibitor also clearly reduced hsp27 expression in VO fed fish. Indicating a p38 MAPK feedback loop, LPS significantly inhibited the expression of p38MAPK itself in both diets, while SB202190 increased p38MAPK expression especially in the VO diet group. hsp70 expression was not affected by any treatment or feed composition. There were also differences in p38MAPK protein phosphorylation comparing treatment groups but no obvious difference comparing the two dietary groups. The results indicate that dietary fatty acids have the ability to modify signalling through p38 MAPK which may have consequences for the fish’s ability to handle infections and stress. Signalling through p38MAPK is ligand dependent and affects gene and protein expression differently. [Copyright &y& Elsevier]

This study sought to investigate whether a "natural diet" (mimicking the fatty acid composition of freshwater aquatic insects eaten by salmon parr) during the freshwater (FW) life stage of pre-smolt Atlantic salmon (Salmo salar L.) affected red blood cells and gill fatty acid composition as well as eicosanoid metabolism in gill during smolting at different temperatures. Before being transferred to seawater (SW), salmon parr were fed with a modified (MO) diet containing vegetable oils (rapeseed, palm, and linseed oils) supplemented with eicosapentaenoic acid (EPA) and arachidonic acid (ARA) to completely replace the fish oil (FO). Fatty acid composition in red blood cells and gill tissues was determined before SW transfer and six weeks after. Additionally, the expression of genes associated with eicosanoid metabolism and Na+/K+-ATPase (NKA) activity in salmon gill was examined at different temperatures before SW transfer and 24 h after. The results showed the changes in fatty acid composition, including sum monounsaturated fatty acids (MUFAs), docosahexaenoic acid (DHA), ARA, EPA, and sum n-6 polyunsaturated fatty acids (n-6 PUFA) in both red blood cells and gill tissues at the FW stage were consistent with the fatty acid profiles of the supplied MO and FO fish diets; however sum EPA and DHA composition exhibited opposite trends to those of the FO diet. The proportion of ARA, EPA, and n-6 PUFA increased, whereas sum MUFAs and DHA decreased in the red blood cells and gill tissues of MO-fed fish compared to those fed with the FO diet at FW stage. Additionally, 5-lipoxygenase-activating protein (Flap) expression was downregulated in MO-fed fish prior to SW transfer. During the process of SW transfer at different temperatures, the MO diet remarkably suppressed NKAα1a expression in MO-fed fish both at 12 and 16 °C. The MO diet also upregulated phospholipase A2 group IV (PLA2g4) expression in gills at 8, 12, and 16 °C, but suppressed phospholipase A2 group VI (PLA2g6) expression in gills at 12 °C compared to FO-fed fish at 12 °C and MO-fed fish at 8 °C. The MO diet also upregulated Cyclooxygenase 2 (Cox-2) expression at 8 °C compared to FO-fed fish and increased Arachidonate 5-lipoxygenase (5-Lox) expression in MO-fed fish at 16 °C compared to both FO-fed fish at 16 °C and MO-fed fish at 8 °C. Our study also determined that both SW transfer water temperatures and diets during the FW period jointly influenced the mRNA expression of PLA2g4 , PLA2g6 , and Lpl , whereas 5-Lox was more sensitive to dietary changes. In conclusion, the MO diet affected the fatty acid composition in gill and in red blood cells. When transferred to SW, dietary ARA supplementation could promote the bioavailability for eicosanoid synthesis in gill mainly via PLA2g4 activation, and potentially inhibit the stress and inflammatory response caused by different water temperatures through dietary EPA supplementation. • Salmon parr were fed with a MO diet only containing vegetable oils supplemented with EPA and ARA. • The MO diet affected fatty acid composition synergistically in gills and red blood cells at FW stage. • A lasting impact of MO diet during FW stage on fatty acid composition after SW transfer. • The MO diet promoted the eicosanoid biosynthesis in gill during SW transfer. • The MO diet inhibited the inflammatory response during smoltification at different temperatures. [ABSTRACT FROM AUTHOR]

The loss of fish in the seawater (SW) phase of Atlantic salmon farming is high, and a major proportion of this loss occurs in the period just after SW transfer. In the current study, we hypothesize that improvements made to the diet during the freshwater (FW) stage affect fish growth, survival and robustness later in the SW stage. To test this, salmon parr were fed five experimental diets in FW at 12 °C. In addition to a commercial-like control diet, fish were fed a diet with changed FA composition aimed to be more like the natural feed of salmon in FW, a diet with increased concentrations of selected AA/N-compounds (methionine, lysine, threonine and taurine), a diet with increased concentrations of methionine and certain B-vitamins (folate, B 12 and B 6) and a final diet combining all of these potential improvements. At the time of SW transfer, the robustness of fish fed the different diets was tested by direct transfer to SW at three different temperatures (8, 12 and 16 °C, without prior acclimation), as well as transfer into open net pens, while fed on a common commercial diet. Growth and proximate composition of the fish did not differ between the diet groups. All diet groups seemed to handle transfer to SW well, and while SW transfer elicited a stress response in the fish, this was not significantly different between diet groups. Fish transferred to SW at 8 °C had higher mortality, reduced mucus layer and increased prevalence of scale loss and wounds, but this applied to all diet groups. Hence, direct transfer to SW at a lower temperature than the fish has been acclimated to cannot be recommended. At the two highest temperatures, there were some differences between the groups in the severity of cataracts. Apart from this, none of the health- or welfare related parameters measured showed any difference between the diet groups, indicating that the control diet was already sufficient. • High loss of farmed salmon after seawater transfer was attempted ameliorated by diet. • The proposed dietary improvements did not affect survival or growth. • The fish did not handle direct transfer from 12 °C freshwater to 8 °C seawater well. • High rearing temperature and probably dietary histidine affected cataract. [ABSTRACT FROM AUTHOR]