The effect of dietary inulin on aerobic bacteria associated with hindgut of Arctic charr (Salvelinus alpinus L.)

The primary aim of the present study was to evaluate the population level of adherent (autochthonous) aerobic and facultative anaerobic bacteria in the hindgut of healthy Arctic charr (Salvelinus alpinus L.) fed dextrin or inulin. This was assessed by the dilution plate technique, and visualized using both transmission and scanning electron microscopy. A population level of 4.8 × 10⁵ adherent bacteria per gram wet mass was found in the hindgut of fish fed a casein‐based diet supplemented with 15% dextrin. However, substituting dextrin with 15% inulin reduced the bacterial population level in the hindgut (3.56 × 10⁴). A total of 217 bacterial isolates were identified by key phenotypical and biochemical characteristics. In addition, 22 strains were also identified by partial sequencing of the 16S rRNA gene. The composition of bacteria colonizing the hindgut of Arctic charr fed dextrin was dominated by the genera Staphylococcus, Pseudomonas, Micrococcus, Psychrobacter glacincola and Streptococcus. However, bacteria colonizing the hindgut of fish fed inulin were dominated by Gram‐positive bacteria of the genera Staphylococcus, Streptococcus, Carnobacterium and Bacillus. While Carnobacterium divergens‐like strains were isolated from charr fed dextrin, Carnobacterium maltaromicus‐like strains were isolated from the hindgut of fish fed inulin. Electron microscopical analysis of hindgut regions confirmed traditional culture‐based microbial analysis as fewer bacterial cells were observed between microvilli and associated with the surfaces of enterocytes of fish fed inulin rather than dextrin.     

Use of 16S rRNA gene sequencing analysis to characterize culturable intestinal bacteria in Atlantic salmon (Salmo salar) fed diets with cellulose or non-starch polysaccharides from soy

Duplicate groups of Atlantic salmon (Salmo salar L.), kept in seawater, were fed fish meal‐based cold‐pelleted diets. Diets with non‐starch polysaccharides (NSP), either cellulose, purified soybean NSP or extruded purified soybean NSP at a dietary level of 100 g kg^?1, were compared with a diet without supplemental NSP and a diet with soybean meal in a 28‐day feeding trial. Isolation and characterizations were limited to culturable bacteria and population levels of aerobic and facultative aerobic heterotrophic autochthonous (adherent) and allochthonous (transient) bacteria present in the mid and distal intestines of Atlantic salmon fed the five different diets estimated using traditional bacteriological techniques. The presence of an autochthonous microbiota was demonstrated using transmission electron microscopy. No significant effects of diet composition were observed on total population levels of culturable bacteria present in the digestive tract, but the study showed that the composition of the gut microbiota (autochthonous or allochthonous) was sensitive to dietary changes. A total of 752 culturable isolates from the intestines were characterized by biochemical and physiological properties. Of these, 188 isolates were further characterized by partial sequencing the 16S rRNA genes. Among these, 146 isolates belonged to 31 phylotypes that were >94% identical to previously described species. However, 42 isolates showed similarity <94% to species available at the National Center of Biotechnology Information. Several of the phylotypes identified in the present study have not been reported previously in the gastrointestinal (GI) tract of fish, including the Gram‐negative bacteria Gelidibacter salicanalis, Pseudoalteromonas elyakovii, Psychrobacter aquimaris, Psychrobacter cibarius, Psychrobacter fozii, Psychrobacter maritimus, Psychrobacter okhotskensis and Psychrobacter psychrophilus. Among the Gram‐positive bacteria identified were Arthrobacter bergeri, Arthrobacter psychrolactophilus, Arthrobacter rhombi, Bacillus pumilus, Bacillus subtilis, Exiguobacterium spp., Microbacterium oxydans, Planococcus maritimus, Sporosarcina ginsengisoli and several bacteria that have been described as unculturable previously. In addition, we identified Carnobacterium inhibens, a lactic acid bacterium that is not frequently isolated from the GI tract of fish. Psychrobacter cibarius was the dominant bacterial species and was isolated from the digestive tract of all fish investigated.     

Effect of Antarctic krillmeal on quality of farmed Atlantic cod (Gadus morhua L.)

Farmed Atlantic cod with a mean weight of 4.8 kg were maintained for 9 weeks in sea cages and fed diets where the dietary fishmeal component was substituted with increasing proportions (0%, 22%, 63% and 100%) of meal from Antarctic krill, Euphausia superba. Wild‐caught cod were included in the study as external control. At termination of feeding, all fish were slaughtered and muscle pH was immediately recorded. The fish were then stored on ice for three days and assessed for muscle pH and objective (skin and fillet colour, and fillet texture) and subjective (sensory evaluation) quality criteria. Replacement of fishmeal with krillmeal in the diets resulted in the skin colour above and below the lateral line to be more red than the control group without krillmeal substitutions, even though this difference was not significant, and with more yellow hue. Additions of krillmeal increased the muscle whiteness and yellow hue compared with the control group and wild fish. There was no difference in red hue between the groups. Muscle pH, texture or sensory attributes were unaffected by dietary inclusion level of krillmeal. Wild‐caught cod deviated in several aspects from the farmed cod. It is concluded that the replacement of fishmeal with Antarctic krillmeal in the diets two months before slaughter did not move the sensory attributes more towards wild fish.     

Tissue fluoride accumulation and kidney lesions in freshwater‐reared Atlantic salmon (Salmo salar) fed with high dietary fluoride concentrations

Antarctic krill (Euphausia superba) and other marine zooplankton may contain high levels of fluoride. The aim of the present experiment was to determine whether dietary fluoride from Antarctic krill at levels similar to the old and the new EU allowable limits in fish feeds (150 and 350 mg kg^?1) would induce kidney lesions in freshwater‐reared Atlantic salmon (Salmo salar). In addition to the diets containing krill, two high‐sodium fluoride (NaF) diets (1500 and 3500 mg kg^?1) were used to investigate the effect on growth, feed intake, faecal excretion of minerals and accumulation of fluoride in various tissues. No major effects on growth or feed intake were observed. A higher proportion of the ingested fluoride was absorbed in salmon fed with the NaF diets compared with fish fed with krill shell diets. Fluoride accumulated in liver, kidney and especially bone. Faecal excretion of calcium and magnesium was higher for the NaF‐fed fish compared with fish fed with the control and krill shell diets, whereas the levels of these minerals in plasma were unaffected. Dietary fluoride from krill shells did not induce kidney lesions. One‐third of the salmon fed with the highest NaF diet showed signs of crystal formation within the distal tubules and/or collecting ducts in the kidney.     

The effect of diet and environmental temperature on the faecal microbiota of farmed Tasmanian Atlantic Salmon (Salmo salar L.)

We investigated the changes in the faecal microbial population of the hindgut of farmed Atlantic salmon fed two commercial diets (A and B) over a 10‐month period. At each sampling round, hindgut contents of 40 fish (20 per diet) were pooled according to diet and cultivated on selective agar. The functional status of all microbiota was assessed against 48 substrates using the PhPlate generalized microplate, which also yielded a metabolic capacity (MC) value for each microbiota. Virulence gene profiles of each microbiota were also investigated. The number of different bacterial groups between two diets varied slightly and increased as water temperature peaked at 18.5°C during summer. This however, was associated with an increase in Vibrio numbers and a decrease in lactic acid bacterial numbers. A shift in the functional status of gut microbiota was observed as temperature increased, which was coupled with a decrease in the MC‐value of the microbiota in both diets. Of the 35 virulence genes tested from total DNA extract of the microbiota, only cdt and east1 were detected. Results indicate that temperature is the main driver of changes to the gut microbiota in farmed Tasmanian Atlantic salmon receiving these two diets.     

Growth, feed conversion and chemical composition of Atlantic salmon (Salmo salar L.) and Atlantic halibut (Hippoglossus hippoglossus L.) fed diets supplemented with krill or amphipods

The effects of partial replacement of fish meal (FM) with meal made from northern krill (Thysanoessa inermis), Antarctic krill (Euphausia superba) or Arctic amphipod (Themsto libellula) as protein source in the diets for Atlantic salmon (Salmo salar L.) and Atlantic halibut (Hippoglossus hippoglossus L.) on growth, feed conversion, macro‐nutrient utilization, muscle chemical composition and fish welfare were studied. Six experimental diets were prepared using a low‐temperature FM diet as control. The other diets included northern krill where 20, 40 or 60% of the dietary FM protein was replaced with protein from northern krill, and two diets where the FM protein was replaced with protein from Antarctic krill or Arctic amphipod at 40% protein replacement level. All diets were iso‐nitrogenous and iso‐caloric. Atlantic salmon grew from 410 g to approximately 1500 g during the 160 day experiment, and Atlantic halibut grew from 345 g to 500–600 g during the 150 day experiment. Inclusion of krill in the diets enhanced specific growth rate in salmon, especially during the first 100 days (P < 0.01), and in a dose–response manner in halibut for over the 150 day feeding period (P < 0.05). Feed conversion ratio did not differ between dietary treatments, and no difference was found in dry matter digestibility, protein digestibility and fish muscle composition. Good growth rates, blood parameters within normal ranges and low mortalities in all experimental treatments indicted that fish health was not affected either Atlantic salmon or Atlantic halibut fed the various zooplankton diets.     

Hydrolysed fish protein concentrate (FPC) reduces viscera mass in Atlantic salmon (Salmo salar) fed plant‐protein‐based diets

Atlantic salmon fed diets devoid of fishmeal but added 0.5 g  kg^?1 fish protein concentrate (FPC) showed reduced growth and lipid deposition without affecting protein accretion as compared to fish fed a fishmeal‐based control diet. The aim of the current study was to assess whether higher inclusion of FPC improved the growth and lipid deposition of Atlantic salmon (initial body weight 380 g) fed high plant protein diets. Quadruplicate groups of fish were fed diets containing 200 g kg^?1 fishmeal of which was replaced with FPC (150, 112, 75, 38 and 0 g kg^?1) for a period of 79 days. The rest of the diet protein was a mixture of plant proteins. The lipid source used was fish oil. A fishmeal‐based diet was included as a positive control for growth performance. None of the test diets differed from the positive control‐fed fish in voluntary feed intake, growth performance or nutrient accretion. Thus, the test diets were found appropriate to assess the effect of FPC inclusion. Replacement of fishmeal with increasing concentration of FPC did not affect voluntary feed intake (P = 0.56), but growth performance decreased (P = 0.02) resulting in an increased feed conversion ratio (P = 0.003). Viscerosomatic index decreased as diet FPC inclusion increased (P = 0.012) without affecting the dress out weight (P = 0.08). Thus, the apparently improved growth in fish fed the diets with the low FPC inclusion was because of a higher visceral mass. Possible reasons for the reduced visceral mass following addition of FPC to high plant protein diets are discussed.     

Short‐term effects of dietary soybean meal and lactic acid bacteria on the intestinal morphology and microbiota of Atlantic salmon (Salmo salar)

The aim of this study was to evaluate the role of the intestinal microbiota in soybean meal enteritis. Three groups of Atlantic salmon (Salmo salar) were fed for 35 days with different diets: a control diet (CD) containing 510 g kg^?1 fishmeal, diet 1 (D1) containing 378 g kg^?1 of soybean meal and diet 2 (D2) containing D1 supplemented with two viable lactic acid bacteria (LAB). As expected, the fish fed with D1 showed clear signs of distal intestinal inflammation during the study compared with the fish fed CD. For the fish fed with D2, the addition of LAB diminished the inflammation at day 28, but did not abolish it. Microbiological analysis demonstrated that specific bacterial groups were not correlated with the development of enteritis, but were correlated with the three diets. Microbacterium, Pseudomonas, Lactococcus lactis sp. cremoris and Aeromonas VIa were correlated with the CD, Aeromonas VIb and Sporosarcina aquimarina were correlated with D1, and Alcaligenes, Acinetobacter, L. lactis sp. lactis and Carnobacterium maltaromaticum were correlated with D2. Shewanella was not affected by the diet and was present in all fish intestines. Our study suggests that LAB may modulate intestinal inflammation; however, the role of the microbiota in the aetiology of enteritis in fish still requires further study.     

The effect of low inclusion levels of Antarctic krill (Euphausia superba) meal on growth performance,apparent digestibility and slaughter quality of Atlantic salmon (Salmo salar)

Two trials with Atlantic salmon (Salmo salar) were conducted to evaluate the potential of krill meal to improve feed intake. In the first experiment, after transfer to sea water, salmon smolts were fed diets added 75 or 150 g kg^?1 Antarctic krill meal in substitution for fish meal for 13 weeks. The apparent digestibility coefficient for crude protein and the majority of the amino acids was significantly lower in the feeds added krill meal (around 83.5%) than in the control diet (84.9%), whereas the digestibility of crude lipids, dry matter and energy was not significantly different among the three diets. Krill meal addition resulted in higher feed intake, which led to higher growth rates and final body weights. In the second experiment, large salmon were fed a diet containing 100 g kg^?1 krill meal for 6 weeks before slaughter. Their feed intake and growth performance were assessed, and fillet and visceral fat contents were measured. Salmon fed the 100 g kg^?1 krill meal diet tended to eat more, resulting in significantly increased growth rates, when compared to control fish. Fish fed krill meal also had a significantly lower condition factor.     

Fillet Quality and Processing Attributes of Postsmolt Atlantic Salmon,Salmo salar,Fed a Fishmeal‐free Diet and a Fishmeal‐based Diet in Recirculation Aquaculture Systems

Many studies have evaluated the adequacy of alternate ingredient diets for Atlantic salmon, Salmo salar, mainly with focus on fish performance and health; however, comprehensive analysis of fillet quality is lacking, particularly for salmon fed these diets in recirculation aquaculture systems (RAS). To this end, a study was conducted comparing fillet quality and processing attributes of postsmolt Atlantic salmon fed a fishmeal‐free diet (FMF) versus a standard fishmeal‐based diet, in replicate RAS. Mean weight of Atlantic salmon fed both diets was 1.72 kg following the 6‐mo trial and survival was >99%. Diet did not affect (P > 0.05) processing and fillet yields, whole‐body proximate composition(fat, moisture, protein), fillet proximate composition, cook yield, fillet texture, color, or omega‐3 fatty acid fillet content, including eicosapentaenoic acid and docosahexaenoic acid levels. Whole‐body ash content was greater in salmon fed the FMF diet. The FMF diet resulted in a wild fish‐in to farmed fish‐out ratio of 0:1 per Monterey Bay Aquarium's Seafood Watch criteria due to its fishmeal‐free status and use of lipids from fishery byproduct. Overall, fillet quality and processing attributes were generally unaffected when feeding a diet devoid of fishmeal to postsmolt Atlantic salmon cultured in RAS. [Correction added on 7 September 2017, after first online publication: the P value in Abstract has been changed from “P < 0.05” to “P > 0.05”.].     

Fluoride retention of Atlantic salmon (Salmo salar) fed krill meal

The present experiment was performed to study how fluoride from krill meal enriched muscle, whole fish and bone of adult Atlantic salmon (Salmo salar) reared in sea water. Atlantic salmon (mean weight 0.5 kg) were divided into four triplicate groups and fed a commercial fish meal based diets with 0, 100, 200 and 300 g krill kg^?1 feed, respectively, for 12 weeks. The fluoride concentrations in the experimental feeds were analysed to be 18, 132, 235 and 358 mg kg^?1, respectively. Growth, mortality and feed efficiency were recorded through the experiment. Fluoride concentration was measured in muscle, whole‐body, and bone initially and after 12 weeks of feeding. The fluoride concentrations in the samples were determined by alkali fusion and fluoride ion‐selective electrode. Growth, mortality and feed efficiency ratio were not affected by the dietary treatments. The results showed that fluoride concentration in muscle, whole body and bone were not affected by the dietary fluoride level. The fluoride concentration in the tissues showed great variation among replicates of the group given the same diet. Fillets of the fish varied between 0.3 and 1.4 mg fluoride kg^?1 wet weight, whereas the whole‐body concentration of fluoride varied between 3.3 and 6.1 mg kg^?1 wet weight and the fluoride bone concentration varied between 5.8 and 7.2 mg kg^?1 fresh weight. These results suggest that Atlantic salmon are highly tolerant of dietary fluoride given as krill meal with concentration of fluoride up to 350 mg kg^?1 diet, and that accumulation of fluoride from feeding diets containing krill meal does not lead to tissue accumulation in the fish, at least over a short period of time.     

The effects of soybean, linseed and marine oils on aerobic gut microbiota of Arctic charr Salvelinus alpinus L. before and after challenge with Aeromonas salmonicida ssp. salmonicida

Populations of heterotrophic bacteria present in the hindgut region of Arctic charr Salvelinus alpinus L. fed dietary soybean, linseed and marine oils before challenge with Aeromonas salmonicida ssp. salmonicida and marine oil after challenge were estimated using the dilution plate technique. There were differences in bacterial composition between the rearing groups before and after challenge, as well as interindividual variations. For example, carnobacteria were only isolated from the hindgut region of fish fed soybean oil and linseed oil before challenge, whereas Carnobacterium spp. and Carnobacterium funditum‐like species were isolated from fish fed the same oils after challenge. Three non‐motile Aeromonas spp. were isolated from infected fish fed marine oil. One of these isolates was identified as identical to A. salmonicida ssp. salmonicida used in&the challenge test by microbial fingerprinting (amplified fragment length polymorphism). Electron microscopic examinations of hindgut regions demonstrated substantial numbers of bacterial cells associated with enterocytes, but bacterial colonization of the enterocyte surface varied between different rearing groups. The potential of bacteria found associated with the hindgut region to inhibit the fish pathogens A. salmonicida, Vibrio salmonicida and Vibrio anguillarum differed between rearing groups.     

Comparative effects of dietary Na+-lactate on Arctic char, Salvelinus alpinus L., and Atlantic salmon, Salmo salar L.

Arctic char, Salvelinus alpinus L., and Atlantic salmon, Salmo salar L., were fed a commercial diet with or without supplementation of 1.5% Na⁺-lactate. Dietary Na⁺-lactate enhanced growth of Arctic char, while that of salmon was unaffected. Dietary ¹⁴C-lactate was retained for significantly longer in the stomach of Arctic char than that of Atlantic salmon. Changes in intestinal cholytaurin hydrolase activity, a bacterially produced enzyme, may indicate that dietary lactate affects the intestinal microbiota of Arctic char but not that of Atlantic salmon. Analysis of bile acids of char showed that dietary Na⁺-Iactate influenced neither intestinal nor gallbladder bile acid composition. Although Arctic char possesses the classical entero-hepatic circulation pathway, no extra loss of bile acids from the fish was observed. It is concluded that the retention time of diet in the stomach of char is significantly longer than that of salmon. This may increase the antibacterial action of lactate in the former, favouring the colonization of lactic-acid-tolerant bacteria in the intestinal tract, some of which produce the enzyme cholytaurin hydrolase.     

Effect of intraperitoneal injection of immunostimulatory substances on allochthonous gut microbiota of Atlantic salmon (Salmo salar L.) determined using denaturing gradient gel electrophoresis

The allochthonous microbiota in the proximal and distal intestine was investigated in three groups of Atlantic salmon (Salmo salar L.) fed a commercial diet and intraperitoneally injected with (a) phosphate‐buffered saline (control), (b) lipopolysaccharide (LPS) from the fish pathogenic bacteria, Aeromonas salmonicida ssp. salmonicida, and (c) laminaran [β‐(1,3)‐d ‐glucan]. Denaturing gradient gel electrophoresis (DGGE) of the hyper variable V3 region was used to present the microbiota in different experimental groups. Sequencing and phylogenetic analysis of excised DGGE bands suggested that an intraperitoneal injection of LPS from A. salmonicida affects the allochthonous gut bacteria of Atlantic salmon to some extent, as Aeromonas enteropelogenes, Aeromonas veroni, Psychrobacter, Lactobacillus letvazi, Lactobacillus satsumensis, Pantoea, swine manure bacterium and several uncultured bacteria were unique for this group. On the other hand, the bacterial diversity of the allochthonous microbiota did not seem to be affected by injection of β‐(1,3)‐d ‐glucan. Sequences of this experimental group were most closely related to A. enteropelogenes, uncultured Escherichia and Lactobacillus aviarius ssp. aviarius.     

Growth of Atlantic Salmon,Salmo salar,Fed Diets Containing Barley Protein Concentrate

A 16-week feeding trial was conducted with juvenile Atlantic salmon (123 g/fish initial weight) to determine the effects of adding either 11% or 22% barley protein concentrate (BPC) to the diet compared to salmon fed a reference diet (0% BPC) containing mostly fishmeal. The fish were stocked into twelve 0.264 m³ tanks, providing 40 fish per tank with an average density of 18.55 kg/m³. Temperature was maintained at ?12°C and salinity at ?2 ppt using ground water during the study. There were no significant differences in specific growth rate (0.62 basal, 0.60 11% BPC, 0.72 22% BPC), feed conversion (1.06, 1.37, 1.09), weight gain (124.3 g, 113.5 g, 155.3 g), proximate composition of the salmon for lipid (8.4%, 7.8%, 9.8%), protein (17.3%, 17.1%, 18.6%), moisture (71.4%, 72.3%, 69.4%), and ash (6.5%, 6.1%, 6.9%) among any of the treatment groups. Salmon fed the diet containing 22% barley protein concentrate, however, had significantly greater energy retention (34%) compared to the fish fed the other diets (23%–28%). The results of this study indicate that barley protein concentrate is an acceptable feed ingredient for Atlantic salmon that should provide performance similar to fishmeal at inclusion rates below 22%.     

Demonstration of salmon farming as a net producer of fish protein and oil

To date aquaculture’s reliance on dietary marine sources has been calculated on a fish weight‐to‐weight basis without considering the absolute amounts of nutrients but this approach neglects the often considerable differences in the nutritional value of fish. We propose simple nutrient‐to‐nutrient‐based dependency measures that take into account these nutritional differences. In the first study reported here, individually tagged Atlantic salmon (Salmo salar) were reared in seawater supplied tanks with feed collection facilities. In the second, commercial net pens were used to grow over 200 000 fish. For both studies, a low marine ingredient feed containing approximately 165 g kg^?1 fishmeal was compared to a control feed (approx 300 g kg^?1 fishmeal) whilst fish oil inclusion was less markedly reduced. The low marine feeds supported similar growth and feed efficiency compared to the control feeds. With the low marine ingredient feeds, the weight of salmon protein and lipid produced through growth exceeded the weight of marine protein and lipid consumed by the fish meaning that salmon farming can be a net producer of fish protein and oil. The amount of n‐3 long‐chain polyunsaturated fatty acids deposited was sufficient to meet current recommendations from human health organizations.     

Protein from Northern krill (Thysanoessa inermis), Antarctic krill (Euphausia superba) and the Arctic amphipod (Themisto libellula) can partially replace fish meal in diets to Atlantic salmon (Salmo salar) without affecting product quality

The main objective of the present study was to evaluate the effect of using three different crustacean meals (Tysanoessa inermis, Euphausia superba, Themisto libellula) on product quality of Atlantic salmon (Salmo salar L.). In order to do this, a total of six iso‐protein, iso‐lipid and iso‐carotenoid diets were prepared. Two experimental diet series were prepared. In the first series, a control feed (K0) was compared with diets where 20%, 40% and 60% of the fish meal protein were replaced with protein from Northern krill T. inermis (K20, K40 and K60, respectively). In the second series, control feed (K0) was compared with diets where 40% of the dietary protein was replaced by protein from T. inermis (K40), Antarctic krill E. superba (AK40) and the Arctic amphipod T. libellula (AMP40). The salmon groups were fed the various diets for 160 days and the average weight of the fish increased from 410 g to around 1500 g. Fish given diets containing krill displayed a general better growth compared with the ones given pure fish meal diet. Replacing fish meal protein with protein from the crustacean sources had, in general, only minor effects on the flesh quality measured both by technical and sensory methods. However, some significant effects were noted. Postmortem muscle pH was generally lower (P < 0.05), for K20, K40, AMP40 in fish fed crustacean diets compared with those receiving the control diet. Increasing the replacement level of non‐fish meal protein from Northern krill (K20, K60) significantly reduced the rigor contraction. Fish given K20 had a slightly firmer meat texture, measured as resistance to post‐rigor compression, especially when compared with K60 (P < 0.05). Fish from the K20 and AMP40 groups had a deeper red flesh coloration [both light reflection (A*‐value and chroma) and flesh astaxanthin concentration] than fish fed K0 and higher inclusions of krill meal. The groups with the highest astaxanthin flesh content also showed the best growth and had the highest feed intake. Finally, a sensory panel analysis differed slightly from the technical measurements in that K0, rather than K20 was given the highest score for hardness and colour. In comparison with K0, AK40 got the lowest salty taste and hardness scores from the panellists relative to the control fish (P < 0.05). Despite minor effects on the present quality measures, it is concluded that meal from three different crustacean species can successfully replace fish meal up to 60% with Northern krill, and 40% of Antarctic krill and amphipod meal of dietary proteins.     

Acute stress alters the intestinal lining of Atlantic salmon, Salmo salar L.: An electron microscopical study

Groups of Atlantic salmon (Salmo salar L.) in feeding (guts filled with faeces) or fasted (three days of diet deprivation) states were subjected to 15 minutes of acute stress. Blood samples and intestinal tissue were collected and prepared for chemical and ultrastructural analyses at intervals post stress until 53 h of recovery. Subjecting fish to acute stress led to significant alterations of the ultrastructure of the enterocytes lining the gastrointestinal tract (GI tract). The most notable effect was substantial damage to the intercellular junctional complexes in midgut regions. These effects appeared within the first hour after stress, were maintained for at least 12 h and were more pronounced in fed than fasted fish. In contrast, hindgut was influenced less by stress and damage was rarely observed. Stress also influenced fish intestinal microbiota. Adherent bacteria decreased in both midgut and hindgut of stressed fish, and this was accompanied by a significant increase in the bacterial contents of faeces. It is suggested that this was due to the sloughing off of mucus eliminating existing microflora and allowing remaining bacteria (also pathogenic) in the gut lumen to colonize the surface of the enterocytes. Although blood haematocrit and plasma cortisol increased following stress, the response appeared to be greater in fasted fish. There were also significant differences in carbohydrate metabolism. While liver glycogen stores were depleted in fasted fish following the mobilization of glucose into plasma, liver glycogen was never depleted in fed fish. As a consequence, plasma glucose levels remained high for more than 12 h of recovery. In fed fish, plasma lactate was also higher than in fasted salmon, and the clearance rate appeared slower. Acute stress induced oxidative stress, as measured through plasma malondialdehyde, but the effect was marginal and nonsignificant. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Alternative dietary protein sources for Atlantic salmon (Salmo salar L.) effect on intestinal microbiota,intestinal and liver histology and growth

The present study has addressed the issue of dietary alterations on gut microbiota in fish by investigating modulation of the allochthonous and autochthonous bacterial gut community of Atlantic salmon following feeding with pea protein concentrate, soy protein concentrate, extracted sunflower, poultry by‐product and feather meal. The results revealed that total allochthonous and total autochthonous bacteria isolated from the distal intestine (DI) were affected by dietary modulations. Total autochthonous bacteria in proximal intestine remained unaffected. Corynebacteriaceae dominated the allochthonous bacteria and Lactobacillaceae dominated the autochthonous bacteria community in proximal and DI indicating that the most abundant groups are not shifted by alternative proteins. Some variations were observed between the bacterial communities as result of dietary manipulations. Alternative protein sources resulted in no severe signs of enteritis. Vacuolization of the liver was unaffected by diet. Furthermore, addition of plant materials significantly affected production of acetic acid. Weight of pyloric‐, proximal‐ and DI was influenced by diet, especially feather meal. Growth performance was significantly affected by inclusion of feather meal to the diet. In conclusion the results showed that pea protein concentrate and poultry by‐products are suitable protein replacements for Atlantic salmon and that soy protein concentrate is a potential candidate.