Proteinase-activated receptor-2: two potential inflammatory mediators of the gastrointestinal tract in Atlantic salmon

Proteinase-activated receptor 2 (PAR-2), activated by trypsin and other serine proteinases, is a key initiator of inflammatory responses in the intestine of mammals. Atlantic salmon fed diets with standard qualities of soybean meal (SBM) show enteritis of the distal intestine as well as increased activity of trypsin in both luminal contents and wall tissue. Luminal trypsin activity may possibly be involved in immune related disorders of the intestine also in Atlantic salmon via activation of PAR 2. In the present study our aim was to investigate if PAR-2 play a role in SBM induced enteritis. We performed multiple alignments based on nucleic acid sequences of PAR-2 from various animals available from public databases, and designed primers for use in cloning of the Atlantic salmon PAR-2 transcript. We further cloned and characterized the full length sequence of Atlantic salmon PAR-2 and investigated the expression in both early and chronic stages of SBM induced enteropathy. Two full length versions of PAR-2 cDNA were identified and termed PAR-2a and PAR-2b. Expression of the two PAR-2 transcripts was detected in all 18 tissues examined, but most extensively in the intestine and gills. A significant up-regulation in the distal intestine was observed for the PAR-2a transcript after 1 day feeding diets containing SBM. After 3 weeks of feeding, PAR-2a was down-regulated compared to the fish fed control diets. These findings may indicate that PAR-2a participates in inflammatory responses in both the early and later stages of the SBM enteropathy. In the chronic stages of the enteropathy, down-regulation of PAR-2a may indicate a possible desensitization of the PAR-2a receptor. Expression of PAR-2b was not altered in the first 7 days of SBM feeding, but a significant up regulation was observed after 3 weeks, suggesting a putative role in chronic stages of SBM induced enteritis. The expression differences of the two PAR-2 transcripts in the feed trials may indicate that they have different roles in the SBM induced enteritis

Consistent changes in the intestinal microbiota of Atlantic salmon fed insect meal diets

Background Being part of fish's natural diets, insects have become a practical alternative feed ingredient for aquaculture. While nutritional values of insects have been extensively studied in various fish species, their impact on the fish microbiota remains to be fully explored. In an 8-week freshwater feeding trial, Atlantic salmon (Salmo salar) were fed either a commercially relevant reference diet or an insect meal diet wherein black soldier fly (Hermetia illucens) larvae meal comprised 60% of total ingredients. Microbiota of digesta and mucosa origin from the proximal and distal intestine were collected and profiled along with feed and water samples. Results The insect meal diet markedly modulated the salmon intestinal microbiota. Salmon fed the insect meal diet showed similar or lower alpha-diversity indices in the digesta but higher alpha-diversity indices in the mucosa. A group of bacterial genera, dominated by members of the Bacillaceae family, was enriched in salmon fed the insect meal diet, which confirms our previous findings in a seawater feeding trial. We also found that microbiota in the intestine closely resembled that of the feeds but was distinct from the water microbiota. Notably, bacterial genera associated with the diet effects were also present in the feeds. Conclusions We conclude that salmon fed the insect meal diets show consistent changes in the intestinal microbiota. The next challenge is to evaluate the extent to which these alterations are attributable to feed microbiota and dietary nutrients, and what these changes mean for fish physiology and health.publishedVersio

A comprehensive transcriptional body map of Atlantic salmon unveils the vital role of the intestine in the immune system and highlights functional specialization within its compartments

The intestine is a barrier organ that plays an important role in the immune system of Atlantic salmon. The immune functions are distributed among the diffuse gut lymphoid tissue containing diverse immune cells, and other cell types. Comparison of intestinal transcriptomes with those of other organs and tissues offers an opportunity to elucidate the specific roles of the intestine and its relationship with other parts of the body. In this work, a meta-analysis was performed on a large volume of data obtained using a genome-wide DNA oligonucleotide microarray. The intestine ranks third by the expression level of immune genes after the spleen and head kidney. The activity of antigen presentation and innate antiviral immunity is higher in the intestine than in any other tissue. By comparing transcriptome profiles, intestine shows the greatest similarity with the gill, head kidney, spleen, epidermis, and olfactory rosette (descending order), which emphasizes the integrity of the peripheral mucosal system and its strong connections with the major lymphoid organs. T cells-specific genes dominate among the genes co-expressed in these tissues. The transcription signature of CD8+ (86 genes, r > 0.9) includes a master gene of immune tolerance foxp3 and other negative regulators. Different segments of the intestine were compared in a separate experiment, in which expression gradients along the intestine were found across several functional groups of genes. The expression of luminal and intracellular (lysosome) proteases is markedly higher in pyloric caeca and distal intestine respectively. Steroid metabolism and cytochromes P450 are highly expressed in pyloric caeca and mid intestine while the distal intestine harbors genes related to vitamin and iron metabolism. The expression of genes for antigen presenting proteins and immunoglobulins shows a gradual increase towards the distal intestine.publishedVersio

Potential of insect-based diets for Atlantic salmon (Salmo salar)

In the present study, we aimed to assess the effect of dietary insect meal (IM) and insect oil (IO) on growth performance, body composition and nutrient digestibility of freshwater reared Atlantic salmon. The IM and IO were produced from black soldier fly larvae (Hermetia illucens, L.; BSF) that had been grown on (1) media containing organic waste streams, or on (2) media partially containing seaweed (Ascophyllum nodosum). The feeding trial of the current study followed a factorial 2 × 3 way-ANOVA experimental design with six dietary groups of Atlantic salmon fed diets with insect-derived ingredients for 8 weeks. A typical industrial diet, with protein from fish meal and soy protein concentrate (SPC) (50:50) and lipids from fish oil and vegetable oil (33:66), was fed to a positive control group. Five experimental diets were formulated, where 85% of the dietary protein was replaced by IM and/or all the vegetable oil was replaced by IO (IM from insects grown on media 1, IO from insects grown on either media 1 (IO1) or media 2 (IO2)). Replacing the dietary fish meal and SPC with insect protein significantly reduced the apparent digestibility coefficients (ADC) of protein, lipid and all amino acids investigated, though remained highly digestible. There were, however, only small differences due to protein or lipid source in growth performance, and no effects of insect ingredients on feed intake or feed conversion ratio. Inclusion of IM-based diets significantly increased both hepatosomatic index and visceral somatic index of Atlantic salmon. Proteinase activity in the intestine was not affected by dietary inclusion of BSF larvae meal, while leucine aminopeptidase activity was lower in fish fed with insect ingredients than the control group. Whole-body protein, lipid, amino acids and minerals contents were not affected by protein or lipid source. In general, this study showed that protein meal and oil from BSF larvae hold a great potential as a source of nutrients for Atlantic salmon.publishedVersio

Optimization of the Balance between Protein, Lipid and Carbohydrate in Diets for Lumpfish (Cyclopterus lumpus)

The aim of this study was to investigate how the dietary balance between protein, lipid, and carbohydrate affects growth, welfare, and health with focus on immune responses, in lumpfish of body weight ranges 1.7-10 g and 15-50 g. A three-component mixture design, with simultaneous variation of the three macronutrients was applied. Growth, tissue and plasma nutrient composition, welfare, cataract frequency, and immune responses in leukocytes isolated from the fish head kidney were studied. Most responses were linear and driven by dietary lipid level, with minor effects of carbohydrate and protein. The growth in 1.7-10 g fish followed a special cubic model with no clear optimum. The fastest growth in the 15-50 g fish was obtained with the diet containing 55% protein, 17% fat, and 6% carbohydrate. The high dietary levels of lipid also gave high lipid levels in the tissues and a trend of improved welfare score, but there were no effects on cataract. The diets with minimum and maximum protein gave suboptimal immune responses in isolated head kidney cells of the 15-50 g fish, while the fastest growing fish showed normal immune responses. We propose that diets for lumpfish from 10-50 g body weight should contain approximately 55% protein, minimum 10% lipid, and maximum 10% carbohydrate. These diets will also be suitable for lumpfish of 1.7-10 g.publishedVersio

Risk assessment of methylsulfonylmethane (MSM). Opinion of the Norwegian Scientific Committee for Food and Environment

acceptedVersio

Psalms for Organ, Volume 12: Ramoth

Pages, 21-24 in Psalms for Organ, Volume 12

Soya saponins and prebiotics alter intestinal functions in Ballan wrasse (Labrus bergylta)

A 5-week feeding trial was conducted in the cleaner fish Ballan wrasse (Labrus bergylta) for a better understanding of the basic biology of the intestinal functions and health in this stomach less species. During the trial, Ballan wrasse was fed either a reference diet, the reference diet supplemented with (i) a commercial prebiotic (Aquate™ SG, 0·4 %) expected to have beneficial effects, (ii) soya saponins (0·7 %) expected to induce inflammation or (iii) a combination of the prebiotics and the soya saponins to find a remedy for gut inflammation. Blood, intestinal tissue and gut content from four consecutive intestinal segments (IN1 – IN4) were collected. No significant differences in fish growth were observed between the four dietary groups. Saponin supplementation, both alone and in combination with prebiotics, increased weight index of IN2 and IN3 and decreased blood plasma glucose, cholesterol and total protein. Dry matter of intestinal content and activity of digestive enzymes were not affected by diet. Histomorphological analyses revealed a progressing inflammation with increased infiltration by immune cells particularly into the distal parts of the intestine in fish fed diets with saponins, both alone and in combination with prebiotics. Gene expression profiles obtained by RNA sequencing and quantitative PCR mirrored the histological and biochemical changes induced by the saponin load. The study demonstrated that Ballan wrasse gut health and digestive function may be markedly affected by feed ingredients containing antinutrients.publishedVersio

Risk assessment of melatonin. Opinion of the Norwegian Scientific Committee for Food and Environment

publishedVersio