Defatted microalgal biomass derived from biorefinery can be potential feed ingredients for carnivorous fish. The present study investigated the growth, feed intake:gain and health parameters in Atlantic salmon fed for 84 days with defatted Nannochloropsis oceania as a fishmeal replacer. Fish fed feeds containing the algal biomass (at 10 and 20% inclusion, alga groups) were compared with groups that consumed alga-devoid feeds (control group). The fish that received 20% alga tended to have reduced weight gain and specific growth rate. Condition factor, feed conversion ratio and feed intake of this fish group were significantly different when compared with the control group. Hepatosomatic and viscerosomatic indices, whole body and fillet proximate composition were not affected by the dietary treatments. Digestibility of dry matter, protein, lipid, ash and energy, as well as retention of lipid and energy of the fish that received feed with 20% alga meal were also significantly different from those of the control group. Serum superoxide dismutase activity of the 10% alga-fed fish was significantly higher compared with the control fish. Although alga feeding did not cause any distal intestinal inflammation, the intestinal proteins that were altered upon feeding 20% algal meal might be pointing to systemic physiological disturbances. In conclusion, feeds with 20% alga had a negative effect on feed intake, FCR, lipid and energy retention and health of the fish. The defatted Nannochloropsis oceania can be used at modest inclusion levels, around 10%, without negative effects on the performance of Atlantic salmon.

Microalgal biomass is a potential feed ingredient that can replace fishmeal and ensure sustainability standards in aquaculture. To understand the efficacy of the defatted biomass from the marine microalga, Desmodesmus sp. a 70-day feeding study was performed with Atlantic salmon (Salmo salar) smolts. Three groups of fish (av. wt. 167 g) were offered either a control feed (without the microalga) or the microalga-containing (10/20%) feeds. At the end of the feeding period, the growth indices (condition factor, specific growth rate) and survival of the microalga-fed fish were not significantly different from the respective values of the control fish, but the feed conversion ratios were inferior. The proximate composition of the whole body of salmon from the three groups did not vary significantly. Compared to the control fish, the 10% alga-fed fish had lower lipid content in their filet. The protein and lipid digestibility in the three feeds did not differ significantly, but the digestibility of energy in the 10% alga-feed was significantly lower than that of the control feed. Furthermore, comparison of the distal intestinal proteome of Atlantic salmon revealed that the expressions of Alpha-2-HS-glycoprotein-like (Ahsg), Myosin-11 isoform X1 (My11) and Dihydrolipoyl dehydrogenase, mitochondrial-like (Dld) were altered by the microalgal feeding. Examination of the physiological status of the fish based on the serum antioxidant capacities did not reveal any alga-feed-related differences. Moreover, the expression of the selected immune and inflammatory marker genes and the micromorphological observations did not indicate any aberration in the intestinal health of the microalga-fed fish. It is possible to include 20% of defatted Desmodesmus sp. in the feeds of Atlantic salmon.

The efficacy of a microbial feed additive (Bactocell®) in countering intestinal inflammation in Atlantic salmon was examined in this study. Fish were fed either the additive-coated feed (probiotic) or feed without it (control). After an initial 3-week feeding, an inflammatory condition was induced by anally intubating all the fish with oxazolone. The fish were offered the feeds for 3 more weeks. Distal intestine from the groups was obtained at 4 h, 24 h, and 3 weeks, after oxazolone treatment. Inflammatory responses were prominent in both groups at 24 h, documented by changes in intestinal micromorphology, expression of inflammation-related genes, and intestinal proteome. The control group was characterized by edema, widening of intestinal villi and lamina propria, infiltration of granulocytes and lymphocytes, and higher expression of genes related to inflammatory responses, mul1b, il1b, tnfa, ifng, compared to the probiotic group or other time points of the control group. Further, the protein expression in the probiotic group at 24 h after inducing inflammation revealed five differentially regulated proteins – Calr, Psma5, Trp1, Ctsb, and Naga. At 3 weeks after intubation, the inflammatory responses subsided in the probiotic group. The findings provide evidence that the microbial additive contributes to intestinal homeostasis in Atlantic salmon.

Atlantic salmon was orally intubated with a highly purified β-glucan product (MacroGard®) to study the recognition of the molecule by the receptor genes, the regulation of the downstream signalling genes and global proteins, and the micromorphological changes in the intestine. The β-glucan receptor genes of Atlantic salmon, sclra, sclrb, sclrc and cr3, seem to recognize the molecule, and initiate the downstream ITAM-motif signalling, as evident from the significantly high mRNA levels of ksyk, mapkin2, il1b and mip2a levels. Among the altered proteins, the Apoa4 (involved in carbohydrate and lipid metabolism); Tagln, Actb (uptake of β-glucan); Psma2 (associated with substrate recognition); and Ckt (energy metabolismrelated) were the overexpressed ones. The underexpressed proteins included the Uk114, Rpl9, Ctsb and Lgal that are connected to proliferation, LPS-stimulation, Il1b and lactose recognition, respectively. Furthermore, the mRNA levels of igt and the number of immune cells in the distal intestine were found to increase upon β-glucan uptake by the fish. This study provides some clues on the mechanisms by which the β-glucan evokes response in Atlantic salmon, particularly at the intestinal level.

Intestinal inflammation in farmed fish is a non-infectious disease that deserves attention because it is a major issue linked to carnivorous fishes. The current norm is to formulate feeds based on plant-derived substances, and the ingredients that have antinutritional factors are known to cause intestinal inflammation in fishes such as Atlantic salmon. Hence, we studied inflammatory responses in the distal intestine of Atlantic salmon that received a feed rich in soybean derivatives, employing histology, transcriptomic and flow cytometry techniques. The fish fed on soy products had altered intestinal morphology as well as upregulated inflammation-associated genes and aberrated ion transport-linked genes. The enriched pathways for the upregulated genes were among others taurine and hypotaurine metabolism, drug metabolism—cytochrome P450 and steroid biosynthesis. The enriched gene ontology terms belonged to transmembrane transporter- and channel-activities. Furthermore, soybean products altered the immune cell counts; lymphocyte-like cell populations were significantly higher in the whole blood of fish fed soy products than those of control fish. Interestingly, the transcriptome of the head kidney did not reveal any differential gene expression, unlike the observations in the distal intestine. The present study demonstrated that soybean derivatives could evoke marked changes in intestinal transport mechanisms and metabolic pathways, and these responses are likely to have a significant impact on the intestine of Atlantic salmon. Hence, soybean-induced enteritis in Atlantic salmon is an ideal model to investigate the inflammatory responses at the cellular and molecular levels.

Nutrient digestibility, growth, and mucosal barrier status of fish skin, gills, and distal intestine were studied in Atlantic salmon fed feeds based on marine or plant-derived ingredients. The barrier status was assessed by considering the expression of four mucin genes, five genes that encode antimicrobial proteins, distal intestine micromorphology, and design-based stereology of the midgut epithelium. In addition, the head kidney leukocytes were examined using flow cytometry; to understand the differences in their counts and function. Five experimental feeds containing the main components i) fishmeal and fish oil (BG1), ii) soybean meal (BG2; to induce enteritis), iii) fishmeal as the main protein source and rapeseed oil as the main lipid source (BG3), iv) a mix of plant protein concentrates as the protein sources and fish oil as the lipid source (BG4), and v) plant and marine ingredients in the ratio 70:30 (BG5) were produced for the study. Atlantic salmon with initial weight 72.7 ± 1.2 g was offered the experimental feeds for 65 days. The results revealed that the weights of all fish groups doubled, except for fish fed BG2. Fish fed the BG2 diet had lower blood cholesterol concentration, developed enteritis, had lower expression of muc2 in the distal intestine, and had a compromised barrier status in the intestine. Expression of both the mucin genes and genes that encode antimicrobial peptides were tissue-specific and some were significantly affected by diet. The fish fed BG1 and BG3 had more head kidney lymphocyte-like cells compared to BG5-fed fish, and the phagocytic activity of macrophage-like cells from the head kidney was the highest in fish fed BG1. The intestinal micromorphology and the mucosal mapping suggest two different ways by which plant-based diets can alter the gut barrier status; by either reducing the mucous cell sizes, volumetric densities and barrier status (as noted for BG2) or increasing volumetric density of mucous cells (as observed for BG4 and BG5). The results of the compromised intestinal barrier in fish fed plant ingredients should be further confirmed through transcriptomic and immunohistochemical studies to refine ingredient composition for sustainable and acceptable healthy diets.

Nutrient limited conditions are common in natural phytoplankton communities and are often used to increase the yield of lipids from industrial microalgae cultivations. Here we studied the effects of bioavailable nitrogen (N) and phosphorus (P) deprivation on the proteome and transcriptome of the oleaginous marine microalga Nannochloropsis gaditana. Turbidostat cultures were used to selectively apply either N or P deprivation, controlling for variables including the light intensity. Global (cell-wide) changes in the proteome were measured using Tandem Mass Tag (TMT) and LC-MS/MS, whilst gene transcript expression of the same samples was quantified by Illumina RNA-sequencing. We detected 3423 proteins, where 1543 and 113 proteins showed significant changes in abundance in N and P treatments, respectively. The analysis includes the global correlation between proteomic and transcriptomic data, the regulation of subcellular proteomes in different compartments, gene/protein functional groups, and metabolic pathways. The results show that triacylglycerol (TAG) accumulation under nitrogen deprivation was associated with substantial downregulation of protein synthesis and photosynthetic activity. Oil accumulation was also accompanied by a diverse set of responses including the upregulation of diacylglycerol acyltransferase (DGAT), lipase, and lipid body associated proteins. Deprivation of phosphorus had comparatively fewer, weaker effects, some of which were linked to the remodeling of respiratory metabolism.

New sustainable feed ingredients are a necessity for the salmon aquaculture industry. In this study, we examined the effect of pre-extrusion processing of two microalgae, Nannochloropsis oceanica and Tetraselmis sp., on the growth, fatty acid content in the flesh and health of Atlantic salmon. The fish were fed one of the following five diets for nine weeks: (1) CO: a fish meal-based control (basal) diet, (2) NU: a Nannochloropsis diet, (3) NE: a pre-extruded Nannochloropsis diet, (4) TU: a Tetraselmis diet, and (5) TE: a pre-extruded Tetraselmis diet. The algae-incorporated diets contained 30% of the respective microalgae. Our results showed that the best growth performance was achieved by the CO diet, followed by the NE diets. Feeding of unprocessed Nannochloropsis and Tetraselmis resulted in a significant reduction in enterocyte vacuolization compared to the CO feeding. A significant effect of processing was noted in the fillet fatty acid content, the intestine and liver structure and the expression of selected genes in the liver. The expression of antioxidant genes in both the liver and intestine, and the accumulation of different fatty acids in the fillet and liver of the extruded algae-fed groups, warrants further investigation. In conclusion, based on the short-term study, 30% inclusion of the microalgae Nannochloropsis oceanica and Tetraselmis sp. can be considered in Atlantic salmon feeds.