An examination of the intestinal tract of Atlantic salmon, Salmo salar L., parr fed different varieties of soy and maize

This study was conducted to investigate the long-term effects of feeding plant products from both traditional breeding and from biotechnology on intestinal somatic indices, histology and cell proliferation in first-feeding Atlantic salmon, Salmo salar L. (initial weight 0.21 +/- 0.02 g). A standard fishmeal diet (standard fishmeal) was formulated to contain fishmeal as the sole protein source and suprex maize as the main starch source. Six experimental diets were then developed: two in which some of the fishmeal was replaced with commercially available, genetically modified Roundup Ready full-fat soybean meal (GM-soy) or commercially available, non-GM full-fat soybean meal (nGM-soy) at a level of 12.5% of the total diet, and four diets in which the suprex maize was replaced with two lines of GM-maize (Dekalb 1; D1 and Pioneer 1; P1), both products of event MON810, and their half-sibling non-GM counterparts (Dekalb 2; D2 and Pioneer 2; P2), at a level of 12.1% of total diet. Each diet was fed to fish in triplicate tanks and the experiment lasted for 8 months, during which the fish reached a final weight of 101-116 g. There was no significant effect of diet on the intestinal indices, nor were histological changes observed in the pyloric caeca or mid intestine. In the distal intestine, one of nine sampled fish fed nGM-soy showed moderate changes, two of nine sampled fish fed GM-soy showed changes, one with moderate and one with severe changes, and two of nine fish fed nGM-maize D2 had moderate changes. Using a monoclonal antibody against proliferating cell nuclear antigen (PCNA), cell proliferative responses to the experimental diets were assessed. In fish fed both soy diets, a significantly higher (P < 0.05) cell proliferation response was observed in the distal intestine concomitant with an increased localization of PCNA positive cells along the whole distal intestinal folds. The PCNA response among the nGM-soy group was significantly higher compared with all the other diet groups. In contrast, for fish exposed to dietary maize (type D) compared with fish fed the standard fishmeal, the soy-diets (GM-soy and nGM-soy) and maize (type P), a significantly lower (P < 0.05) cell proliferation response was observed in the distal intestine. Results indicated that the GM plant products investigated in this study, at about 12% inclusion level, were as safe as commercially available non-GM products, at least in terms of their effect on indices and histological parameters of the Atlantic salmon intestinal tract.     

Growth, feed utilization and health of Atlantic salmon Salmo salar L. fed genetically modified compared to non-modified commercial hybrid soybeans

The present paper represents a part of a major scientific effort aiming to reveal possible effects, nutritional or health related, of genetically modified (GM) feed ingredients for Atlantic salmon. For 3 months groups of post‐smolt Atlantic salmon were treated with diets holding 130 g kg¹ of the protein as soybean, one which contained 800 g kg^?1 GM type RR (Roundup Ready^?), and compared with a standard counterpart (commercial hybrid, not isogenic line) analysed to be non‐GM (nGM), and again compared with a standard fishmeal diet without soybean protein. All diets were composed to be within the category ‘compositional equivalent’ and held exactly the same proximate compositions, starch and sugar levels, above requirements for methionine and lysine, equal fatty acid profiles, vitamin, mineral and pigment contents. There was, however, a slight difference in levels of anti‐nutrients between the three diets. The various dietary treatments resulted in more than tripling of fish weight in all groups. In addition no significant differences in feed utilization, whole body, liver and muscle proximate compositions, and no significant differences in muscle fatty acid profiles were measured. The diet without soybean of either type resulted in greater retention of lipid, but equal retention of protein (protein productive value). The relative sizes of liver, kidney, head–kidney and brain were the same in all dietary groups, while the relative size of the spleen showed significant differences between fish fed the genetically modified soy diet compared with fish fed the nGM soybeans. Fish fed the soy diets of either type also showed a somewhat reduced mean erythrocyte cell volume. All other haematological values were equal between diet groups. The detoxification system, measured as glutathione peroxidase (GPx) and lysozyme activities, showed equal values for all groups when measured in plasma and liver or head–kidney. The distal part of the intestine showed reduced sizes as an effect of soybean additions, without any differences between the GM and nGM type. Our results showed high growth, no mortality, haematological values within normal ranges, and efficient and equal responses in the detoxification system. This was a first indication that up to 130 g kg^?1 RR‐soybean protein can safely be used in diets for Atlantic salmon. However, there is still a need to elucidate higher inclusion levels of GM feed ingredients, and why spleen index was reduced, and if this was a long‐ or short‐term effect.     

Alcohol-soluble components in soybeans cause morphological changes in the distal intestine of Atlantic salmon, Salmo salar L.

Standard soybean meal, heat treated to differing degrees and containing varying levels of soybean lectin and trypsin inhibitor, was used to study the effect of soybean-containing diets on the morphology of the distal intestine in Atlantic salmon, Salmo salar L. A reference diet based on fish meal and a fish meal diet mixed with an alcohol extract of soybean meal were used. The fish (145 g) were kept in 27 m³ net pens (200 fish per pen) in sea water with an average temperature of 11 °C for 12 weeks. Whereas a normal morphology of the distal intestine was seen in the fish fed the reference diet, all soybean diets caused alterations in the distal intestine irrespective of heat treatment. Similar changes were observed with the diet which had added alcohol solubles. It is concluded that alcohol-soluble components in the soybeans are responsible for the observed changes. It cannot be determined which of the soybean antinutrients is causing the alterations from the present study. Oligosaccharides and saponines are possible candidates. However, the possibility that unknown antinutrients are affecting the salmon intestine should not be overlooked.     

Dehulled solvent-extracted soybean meal as a protein source in diets for Atlantic salmon, Salmo salar L.

Diets, containing dehulled, solvent-extracted soybean meal in amounts corresponding to 0. 20 and 40% soybean protein of total protein, were fed for 10 months to duplicate groups of Atlantic salmon, Salmo salar L., weighing about 900 g, held in sea water. Fish fed the diet containing 20% soybean protein grew as well as fish fed the diet with high-quality fish meal as the sole protein source, whilst the fish fed the diet containing 40% soybean protein grew significantly less. No significant differences were found for dressing percentage, condition factor or relative liver weight between the dietary treatments. Fish fed the control or the 20% soybean meal (SBM) diets were not significantly different in body fat content, whilst this was significantly lower in the fish fed the 40% SBM diet. There were no significant differences between the dietary treatments in body protein content. Inclusion of SBM seemed to affect neither carcass calcium, carcass zinc, nor plasma calcium, magnesium or potassium. The plasma zinc content increased and plasma iron decreased by inclusion of SBM in the diets, probably reflecting dietary contents of zinc and iron. Liver iron was not affected. Plasma phosphate was significantly decreased by inclusion of SBM in the diet, whilst plasma glucose, total protein and haemoglobin were not affected. For plasma free amino acids, the only significant differences were a decrease in taurine with increasing SBM inclusion and methionine was significantly higher for the SBM groups than for the control groups.     

Effects of diet supplementation of soya‐saponins,isoflavones and phytosterols on Atlantic salmon (Salmo salar,L) fry fed from start‐feeding

A 14‐week trial was conducted to investigate the effects of antinutritional factors (ANFs) commonly present in soybean ingredients, singly and in combination, on Atlantic salmon (Salmo salar L.) fed from start‐feeding. The experimental diets consisted of a negative control fish meal diet (FM), and a positive control diet with 167 g kg^?1 soybean meal inclusion (SBM) and four diets based on the FM diet supplemented with 2 g kg^?1 soya‐saponins (SAP), 1.5 g kg^?1 isoflavones (IFL), 0.3 g kg^?1 phytosterols (PHS) or a mixture of these (MIX). Fish fed the SAP diet showed significantly higher growth performance than those fed FM, while the IFL treatment significantly decreased growth performance of salmon fry. Fish fed the IFL diet had significantly lower maltase activity and higher trypsin activity in proximal intestine than fish fed the FM diet. Histological differences were observed in the liver of fish fed the IFL diet, characterized by reduced size of the hepatocytes. Fish fed the PHS and IFL diets showed the highest frequencies of skeletal deformities among the six treatments. In conclusion, the results indicate that purified isoflavones may negatively affect growth performance, intestinal function, liver metabolism and bone formation of salmon fry.     

Feeding Atlantic salmon Salmo salar L. soybean products: effects on disease resistance (furunculosis), and lysozyme and IgM levels in the intestinal mucosa

Two trials were initiated to investigate the consequences of including various soybean products in diets for Atlantic salmon Salmo salar L. on (1) mortality following infection by Aeromonas salmonicida ssp. salmonicida during a cohabitation challenge, and (2) the lysozyme and IgM content of the intestinal mucosa. Groups of salmon were fed control diets containing fishmeal as the sole protein source (Contr1 and Contr2, respectively), soy concentrate-containing diets (SoyConc1 and SoyConc2, respectively), or diets containing either solvent-extracted soybean meal (SoyMeal, trial 1) or soybean molasses (SoyMol, trial 2), an alcohol extract of soybean meal. Both SoyMeal and SoyMol caused enteritis-like changes in the distal intestine, which were not observed in fish fed the Contr1, Contr2, SoyConc1, or SoyConc2 diets. There were significant differences ( P  < 0.05) in mortality between feeding groups following the A.s. salmonicida challenge: these differences were greatest in fish fed SoyMeal (65.6%), least in fish fed SoyConc1 (60.5%), and intermediate in the fish fed the Contr1 diet (62.9%). The SoyMol diet caused significantly ( P  < 0.0001) increased levels of both lysozyme and IgM in the mid and distal intestinal mucosa. It is concluded that components of soybean meal and soybean molasses cause an inflammatory response in the distal intestine that may lead to increased susceptibility to furunculosis.     

Utilization of genetically modified soybean meal in Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> diets

The utilization of genetically modified soybean meal (GM SBM) was compared with that of non-GM SBM in Nile tilapia. Four experimental diets were formulated to include either non-GM or GM SBM at 34 or 48%, respectively. These diets were fed to juvenile Nile tilapia (49.5 g average weight) for 12 weeks. The uptake of the cauliflower mosaic virus 35S promoter fragment of the GM SBM in fish muscle was examined at 8th and 12th week. After 12th week, fish were fed the non-GM SBM diets to determine the residual span of the incorporated promoter fragment. There was no significant difference in specific growth rate or feed efficiency between GM and non-GM groups at the same inclusion level. A small number of muscles from fish receiving both levels of GM SBM diet were positive for the promoter fragment. Additionally, the promoter fragment was not detected by the second day after changing to the non-GM SBM diets. These results indicate that the utilization of GM SBM was similar to that of non-GM SBM and the promoter fragment was rarely found in fish muscles, suggesting that suitability and safety of GM SBM in Nile tilapia diet were similar to those of non-GM SBM.     

Nutritional, physiological, and histological responses in Atlantic salmon, Salmo salar L. fed diets with genetically modified maize

The objective of this study was to evaluate whether standard fish meal diets prepared with increasing levels of genetically modified (GM; 150 and 300 g kg⁻¹) maize (event MON810^®) as a starch source, showed any nutritional or physiological adverse effects on Atlantic salmon, Salmo salar L. postsmolt. The diets with low or high inclusions of GM maize and its near‐isogenic parental line (nongenetically modified; nGM maize), were balanced with Suprex maize (Reference) to obtain compositional equivalency of diet starch, sugars and all other nutrients. Total starch level in all diets was 160 g kg⁻¹. After 82 days of feeding, fish growth was high in all groups, however fish fed the GM maize showed slight but significant lower feed intake, which was followed by slight but significant lower specific growth rate and final body weights, compared with fish fed nGM maize, none of the groups varied significantly from fish fed the Reference diet. There was no variation in feed conversion ratios (FCR), protein and lipid efficiency ratios (PER and LER), or protein‐ and lipid‐productive values (PPV and LPV) in this study. No significant effect of maize type was detected on apparent digestibility coefficients (ADC) of dry matter, protein or lipid. Hematological analysis and plasma nutrients varied within normal ranges for Atlantic salmon in all diet groups, except for somewhat elevated aspartate aminotransferase (ASAT) values in all groups. Hepatosomatic index (HSI) with values ranging from 1.37 to 1.60, was significantly higher for the high GM maize group compared with the high nGM maize group but not when compared with the Reference diet group. Lowered spleen (SSI) and head‐kidney somatic indices (H‐KSI) were registered when fed GM compared with nGM maize, the Reference treatment was however, equal to both. Distal intestine somatic index (DISI) was significantly higher for GM maize‐fed fish compared with nGM maize‐fed fish, but not significantly different from the Reference diet group. Histological evaluation of the mid‐ and distal intestine, liver, spleen and head‐kidney did not reveal any diet‐related morphological changes. Maltase activities in the mid‐ and distal intestinal tissue homogenates were affected by diet, the fish fed high GM maize having higher activities compared with high nGM maize‐fed fish. Leucine aminopeptidase (LAP) and alkaline phosphatase (AP) activities were not affected by diet. Sodium‐dependent d ‐glucose uptake in brush border membrane vesicles (BBMV) isolated from pyloric caeca of fish fed high GM maize was significantly higher than that found in fish fed the analogous diet with high nGM maize. Based on the present findings, the conclusions made are: Atlantic salmon smolts fed GM maize (event MON810^®), its near‐isogenic parental line and suprex maize (Reference diet), all resulted in high growth rates, ADC and feed utilization. Health, when evaluated by means of mortality (low), normal ranges of blood and plasma parameters, except somewhat elevated ASAT values and minor variations in organ sizes, were considered good in all diet groups. The changes in the glucose transport mechanism and intestinal maltase enzyme activity in the gastrointestinal tract warrant further studies.     

Effects of graded levels of standard soybean meal on intestinal structure, mucosal enzyme activities, and pancreatic response in Atlantic salmon (Salmo salar L.)

Duplicate tanks of c.280 g Atlantic salmon (Salmo salar L.) were fed for 60 days on diets in which fishmeal was substituted with graded levels of extracted soybean meal (SBM) comprising 0%, 10%, 15%, 20%, 25% or 35% of total protein. The effects on feed intake, growth, feed conversion, apparent digestibility and utilization of macronutrients and energy, pathohistological response of the distal intestine (DI), activities of digestive enzymes in the mid and distal intestinal mucosa, and faecal trypsin and plasma insulin concentrations were studied. A negative, dose‐dependent effect of SBM was observed on nearly all performance parameters with a notable exception of feed intake. The lowest SBM inclusion level of 10% resulted in moderate pathohistological changes in the DI. Each subsequent increase in SBM level increased the number of fish displaying severe changes. In contrast to the mid‐intestine (MI), all enzyme activities in the distal intestinal mucosa decreased dose‐dependently with increasing SBM inclusion. Faecal trypsin increased up to an SBM inclusion level of 20% and then levelled off. Plasma insulin increased from 0% to 15% SBM inclusion and then decreased. The results suggest that caution should be exercised in the use of even low levels of extracted SBM in salmon feeds.     

Organs development,gene expression and health of Atlantic salmon (Salmo salar L.) fed genetically modified soybeans compared to the near‐isogenic non‐modified parental line

The present experiment was conducted to study the possible effects of genetically modified (GM), full‐fat soybean meal (FFSBM) from Round‐up Ready^® soybeans compared to its parental, and closest near‐isogenic, non‐modified (nGM) soybean variety, added at moderate (150 g kg^?1) and high (300 g kg^?1) inclusion levels. The fish showed a high specific growth rate (SGR 1.27–1.52), and nearly doubled their body weight (BW), with final weights varying from 1009 to 1110 g. Increased levels of dietary FFSBM, independent of the soy being GM or not, significantly decreased mean values of SGR, thermal growth rate, condition factor, final BW, liver somatic index, lipid efficiency ratio, apparent digestibility coefficients (ADC) of protein and gross energy, liver lipid content and plasma cholesterol, and significantly increased ADC of starch and muscle fatty acid levels of 18:3n‐3, 20:4n‐6, 20:5n‐3 and total n‐3. Increasing dietary GM FFSBM significantly increased feed conversion ratio, and significantly decreased protein efficiency ratio, ADC of lipid and dry matter and plasma triacylglycerol (TAG) levels. Spleen somatic index was significantly larger in fish groups fed GM FFSBM compared to groups fed nGM FFSBM, which might indicate a possible immune response exerted by the GM soybeans. Mean normalized expression of heat shock protein 70 mRNA in distal intestine was significantly up‐regulated while normalized expression of catalase in liver was down‐regulated, in fish fed FFSBM compared to fish fed FM. In conclusion, substituting moderate to high levels of GM Round‐up Ready^® FFSBM in diets for Atlantic salmon and compared to the closest near‐isogenic counterpart available, resulted in many effects independent of the soy being GM or not, but with the notable exceptions of enlarged spleen and lowered plasma TAG.     

Suitability of genetically modified soybean meal in a dietary ingredient for common carp Cyprinus carpio

The effect of genetically modified (GM) soybean meal (SBM) in a feed ingredient on growth performance of common carp was investigated in comparison to nonGM SBM. GM SBM was included at 34 and 48% in two experimental diets that were formulated with fish meal (FM) to obtain approximately 38% protein in diet. Two other experimental diets were formulated to contain the same levels of nonGM SBM. The diets were fed to juvenile common carp (22 g initial mean weight) for 12 weeks. The cauliflower mosaic virus (CaMV) 35S promoter fragment (205 base pairs) of the GM SBM was examined in fish muscle and blood samples at the twelfth week. From the twelfth week, the GM groups were fed with nonGM diets to determine the residual span of the transferred promoter fragment. There was no significant difference in growth and feed performance between GM and nonGM groups at two inclusion levels after 12 weeks. The CaMV 35S promoter fragment was not detected in fish muscles or blood receiving either level of GM SBM diet. The results demonstrated that the availability of GM SBM was similar to that of nonGM SBM and the GM SBM would be a suitable and safe ingredient in feed for common carp.     

Digestive function and intestinal integrity in Atlantic salmon (Salmo salar) fed kernel meals and protein concentrates made from yellow or narrow-leafed lupins

This study assessed the effects of yellow lupin (Lupinus luteus) and narrow-leafed lupin (L. angustifolius) kernel meals and protein concentrates on the gastrointestinal integrity, capacity for digestive hydrolysis, and digestibility of nutrients in Atlantic salmon. A basal diet (FM) was made from fish meal, wheat, and fish oil. Six additional diets were formulated by replacing 30% of the FM diet with lupin kernel meal made from L. l. cv. Wodjil (LKM), L. a. cv. Belara (BKM), and L. a. cv. Myallie (MKM), lupin protein concentrates made from the same L. l. (LPC) and L. a. cv. M (MPC), or extracted soybean meal (SBM). All diets were extruded. Each diet was fed to three groups of 176 g salmon kept in 1 m² tanks with 5.6 °C saltwater for 3 weeks prior to sampling of blood, intestinal organs, digesta, and faeces. Inclusion of lupin meals in the diets resulted in harder and more condensed feed particles. Ulcer-like lesions were observed in the stomach of fish from all feeding groups, and this was worsened by lupin in the diet, but did not appear to be pellet hardness related. No consistent altered morphology was observed in the distal intestine (DI) of fish fed the FM and lupin diets, while the DI of fish fed SBM showed consistent and typical soybean meal-induced pathomorphological changes. Plasma cholesterol was higher when feeding MKM and LKM than when feeding FM, MPC, and LPC, with intermediate levels when feeding BKM and SBM. Feeding LKM and LPC resulted in a higher weight of the GIT when related to body weight. Trypsin activity and bile acid concentration were generally higher in digesta from the pyloric (PI) and mid (MI) intestine when feeding FM and lupin diets than when feeding SBM, while the opposite was seen for trypsin activity in digesta from DI. There were no effects of diet on leucine aminopeptidase (LAP) and maltase activity in PI and MI, but in DI the activity of these brush border enzymes were significantly lowered when feeding SBM. SBM in the diet resulted in watery faeces and lowered apparent digestibility of lipid, but this was not observed when feeding the lupin diets. To conclude, the tested lupin kernel meals and protein concentrates did not alter the intestinal function in Atlantic salmon when included at 30% of the diet. Dietary lupin was, however, involved in the worsening of ulcer-like gastric lesions.     

Changes in immune and enzyme histochemical phenotypes of cells in the intestinal mucosa of Atlantic salmon,Salmo salar L., with soybean meal‐induced enteritis

Extracted soybean meal (SBM) in the diet for Atlantic salmon, Salmo salar L., causes an inflammatory response in the distal intestine. The morphological changes of the epithelial cells and a characterization of the inflammatory cell infiltrate of the distal intestinal mucosa were studied using a panel of enzyme and immunohistochemical markers. The salmon (average body weight 927 g) used in the study were fed either a fishmeal‐based diet (control diet) or a diet in which 30% of the fishmeal protein was replaced with SBM protein (SBM diet). In salmon fed SBM, there were markedly reduced enzyme reactivities in the distal intestinal epithelial cells, both in the brush border [5′‐nucleotidase (5′N), Mg2+‐ATPase, alkaline phosphatase (ALP) and leucine aminopeptidase (LAP)] and in the intracellular structures [alkaline and acid phosphatase, non‐specific esterase (NSE) and alanine aminopeptidase (AAP)]. There appeared to be an increased presence of cells of monocytic lineage, including macrophages, as well as neutrophilic granulocytes and immunoglobulin (Ig) M in the lamina propria of the SBM‐fed fish. The mid intestine showed little response to the diet. The results suggest that toxic/antigenic component(s) of SBM affect the differentiation of the distal intestinal epithelial cells and may help explain the reduced nutrient digestibilities previously reported in salmonids fed extracted SBM.     

Effects of feed processing conditions and acid salts on nutrient digestibility and physical quality of soy‐based diets for Atlantic salmon (Salmo salar)

Two experiments were conducted to investigate effects of feed processing conditions and potassium diformate (KDF) supplementation on apparent digestibility of nutrients in Atlantic salmon (Salmo salar) and physical quality of extruded feed. In Exp. 1, diets with raw or expander pretreated full‐fat soybean meal (FFSBM) at 100 or 120 °C were extruded at 110 °C. Expander pretreatment significantly (P < 0.05) improved the digestibility of arginine, glutamine and tyrosine in Atlantic salmon. The higher digestibility of expander pretreated FFSBM was confirmed in mink (Mustela vison). In Exp. 2, diets with defatted soybean meal (SBM) were extruded at 110, 130 or 150 °C. The results showed that increasing extrusion temperatures significantly (P < 0.05) improved the digestibility of most major nutrients and amino acids in Atlantic salmon. In general, KDF supplementation to FFSBM and SBM diets did not affect digestibility of nutrients in Atlantic salmon or mink. Expander pretreatment and increasing extrusion temperatures increased pellet expansion, while KDF supplementation reduced pellet expansion.     

Nutrient digestibilities and gut function in Atlantic salmon (Salmo salar) fed diets with cellulose or non-starch polysaccharides from soy

Duplicate groups of Atlantic salmon (Salmo salar), kept in saltwater, were fed fish meal based cold-pelleted diets where graded levels of native or extruded non-starch polysaccharides (NSP) from soybeans replaced cellulose, at a total NSP level of 100 g kg^− 1 for 28 days. The study also included a diet where defatted soybean meal (SBM) constituted the NSP at a level of 100 g kg^− 1 and a reference diet without NSP supplementation.The SBM diet resulted in a significant reduction in faecal dry matter content, apparent digestibilities of starch and organic material, and growth, and an increased faecal output of several elements (K, Na, Zn), compared to all the other diets. Morphological changes were only seen in the intestine of the fish fed the SBM, i.e. the diets holding purified soy-NSP did not induce enteritis. When compared to the diet without NSP, cellulose addition increased faecal dry matter, while inclusion of native soy-NSP reduced it. Dry matter in faeces and apparent digestibility of crude protein decreased in a linear manner, while the digestibility of starch and faecal output of K and Na increased linearly when native soy-NSP replaced graded levels of cellulose from 0 to 100 g kg^− 1 feed. When diets with 75 or 100 g kg^− 1 of native NSP and extruded soy-NSP were compared, fish fed native soy-NSP had reduced faecal dry matter, higher digestibility of starch, and increased faecal output of Cu, Fe, and K. Dry matter in faeces and faecal output of Cu was lower for the highest inclusion level, while digestibility of starch and faecal output of Mn and K were higher.In conclusion, soy-NSP was inert compared to the fish meal reference, with respect to nutrient digestibilities and intestinal pathologies, but affected faecal mineral excretion in Atlantic salmon.     

Alcohol soluble components of soybeans seem to reduce fat digestibility in fish-meal-based diets for Atlantic salmon, Salmo salar L.

A commercially available soybean molasses (alcohol extract from soybean meal) was included at levels of 0. 5,10,15 and 20% in a fish meal based diet for Atlantic salmon, Salmo salar L., weighing about 75 g. These inclusion levels supplied dietary alcohol soluble components corresponding to levels that would have been obtained if a standard soybean meal was included at levels up to about 70%. The fish were reared in fresh water at 9^oC and with a minimum of 7 ppm O₂ at outlet, and fed the experimental diets for 14 days before the faeces were stripped. Digestibility coefficients were determined using chromic oxide as an indigestible indicator. The digestibility of fat, and particularly the long-chained, saturated-and monounsaturated fatty acids, were significantly reduced by increasing the inclusion level of soybean molasses, whereas digestibility of dry matter, protein (%N x 6.25) and polyunsaturated fatty acids were not affected significantly. The present results show that alcohol-soluble components of soybeans may be responsible for a significant part of the negative effects of standard soybean meal in diets for Atlantic salmon.     

Time-related changes of the intestinal morphology of Atlantic salmon, Salmo salar L., at two different soybean meal inclusion levels

Soybean meal (SBM) induces enteritis in the distal intestine of Atlantic salmon. The present study assesses the effects of SBM concentrations on the kinetics of the enteritis process. Fish of 300 g, kept at 12 °C, were fed diets with different SBM inclusions: 0%, 10% and 20% SBM for 57 days. Samples of the distal intestine of five fish per treatment were taken for histological and electron microscopical analysis. A semi-quantitative scoring system was used to assess the degree of the morphological changes induced by SBM feeding in the distal intestinal epithelium. The first signs of enteritis appeared earlier in the salmon fed the 20SBM diet than in those fed the 10SBM diet. Thereafter, the condition increased steadily, displaying no signs of recovery. Furthermore, at the lower concentration, the process marking the onset of enteritis began more gradually than at the higher concentration and it displayed a tendency to stabilize after 13–20 days of continuous feeding. Electron microscopy indicated that the endocytosis process was hampered at day 3 of 20SBM and at 7 days of 10SBM. Furthermore, a strong reduction of microvilli was already evident after 7 days of 20SBM feeding, thus indicating a decreased uptake capacity of the distal enterocytes. In addition, transformation and migration of eosinophilic granulocytes was observed, which, in combination with the lysozyme C immunoreactivity supports their protective role during the inflammatory process in the distal gut of Atlantic salmon. It can be concluded that the severity of enteritis and its kinetics are concentration-dependent, showing no signs of recovery during feeding with diets containing SBM.     

Uptake and clearance of dietary DNA in the intestine of Atlantic salmon (Salmo salar L.) fed conventional or genetically modified soybeans

The aim of this study was to examine the presence or absence of dietary transgenic (Roundup Ready^® soybean – RRS^®) and soybean DNA (sRubisco) in the intestinal tract of Atlantic salmon fed either genetically modified (GM) or conventional (non‐GM) soybeans. Uptake of dietary DNA was evaluated in the post gastric intestine (pyloric ceca – PC, mid intestine – MI and distal intestine – DI) after continuous feeding (6 months), feed restriction and re‐feeding using qPCR and in situ hybridization. No transgenic DNA fragments were detected in any of the intestinal samples using event specific primers. Soybean DNA was detected in all segments of the intestinal tissue (PC, MI and DI) and visualized in the cell vacuolar system of the DI in the apex area of the intestinal fold. Dietary DNA was gradually cleared from the intestinal tissues when feed was restricted and could not be detected after 5 days. Re‐feeding resulted in dietary plant DNA uptake after 2 h. The results show that the salmon intestine is able to take up dietary plant DNA shortly after feed intake and that one of the factors affecting uptake and clearance of nucleic acids in the various intestinal segments are the feeding status of the fish.     

Effects of substitution of dietary fish oil with a blend of vegetable oils on liver and peripheral blood leucocyte fatty acid composition, plasma prostaglandin E2 and immune parameters in three strains of Atlantic salmon (Salmo salar)

Duplicate groups of three genetic strains of Atlantic salmon smolts were cultured on diets containing either fish oil (FO) or a blend of vegetable oils (VO). Fatty acid compositions of liver and peripheral blood leucocytes were compared. The effect of different strains and diets on innate immune parameters and plasma prostaglandin E₂ were also measured. Two strains were selected as being either 'fat' or 'lean' in terms of muscle adiposity. The third strain was a commercial stock (MH). Total replacement of dietary FO with VO resulted in reduced docosahexaenoic (DHA; 22:6 n -3) and eicosapentaenoic acids (EPA; 20:5 n -3) in liver, while oleic (18:1 n -9), linoleic (18:2 n -6) and α-linolenic (18:3 n -3) acids were all increased in VO-fed fish. Fatty acid compositions of blood leucocytes showed similar changes. Evaluation of innate immune function showed that in the fat strain, circulating leucocytes were significantly lower in VO fish. The lean strain also had significantly higher serum lysozyme activity than MH fish. Reduced haematocrit was seen in VO lean fish compared with FO lean fish. This study provides evidence of strain-induced differences in liver and leucocyte fatty acid compositions and innate immunity in Atlantic salmon fed either FO- or VO-based diets.