Evaluation of fixation methods for demonstration of Neoparamoeba perurans infection in Atlantic salmon,Salmo salar L., gills

Formaldehyde‐based fixatives are generally employed in histopathology despite some significant disadvantages associated with their usage. Formaldehyde fixes tissue by covalently cross‐linking proteins, a process known to mask epitopes which in turn can reduce the intensity of immunohistochemical stains widely used in disease diagnostics. Additionally, formaldehyde fixation greatly limits the ability to recover DNA and mRNA from fixed specimens to the detriment of further downstream molecular analyses. Amoebic gill disease (AGD) has been reliably diagnosed from histological examination of gills although complementary methods such as in situ hybridization (ISH) and polymerase chain reaction (PCR) are required to confirm the presence of Neoparamoeba perurans, the causative agent of AGD. As molecular techniques are becoming more prevalent for pathogen identification, there is a need to adapt specimen collection and preservation so that both histology and molecular biology can be used to diagnose the same sample. This study used a general approach to evaluate five different fixatives for Atlantic salmon, Salmo salar L., gills. Neutral‐buffered formalin and seawater Davidson's, formaldehyde‐based fixatives commonly used in fish histopathology, were compared to formalin‐free commercial fixatives PAXgene^®, HistoChoice?MB* and RNAlater?. Each fixative was assessed by a suite of analyses used to demonstrate AGD including routine histochemical stains, immunohistochemical stains, ISH and DNA extraction followed by PCR. All five fixatives were suitable for histological examination of Atlantic salmon gills, with seawater Davidson's providing the best quality histopathology results. Of the fixatives evaluated seawater Davidson's and PAXgene^® were shown to be the most compatible with molecular biology techniques. They both provided good DNA recovery, quantity and integrity, from fixed and embedded specimens. The capacity to preserve tissue and cellular morphology in addition to allowing molecular analyses of the same specimens makes seawater Davidson's and PAXgene^® appear to be the best fixation methods for diagnosis and research on AGD in Atlantic salmon gills.     

Tracing the route of Sphaerospora truttae from the entry locus to the target organ of the host, Salmo salar L., using an optimized and specific in situ hybridization technique

Sphaerospora truttae is an important pathogen of Atlantic salmon parr in Scottish aquaculture. To trace the early development of S. truttae and to overcome the common problem of detecting low numbers of cryptic, early myxosporean stages, a DNA-based approach was applied in this study. Specific primers were designed for S. truttae from Atlantic salmon, based on 18S rDNA sequences, obtained from isolated myxosporean spores. These were 5' biotin-labelled and used in an optimized and rapid in situ hybridization (ISH) protocol, which provided a strong and specific signal of the parasite within host tissue sections and, at the same time, minimized structural damage to tissues due to processing. This methodology provided a reliable tool enabling the detection of S. truttae stages down to single cell level. Using ISH the epithelium of the gills was identified as the predominant entry locus of the parasite. By 3 days after infection S. truttae had penetrated the vascular epithelia and thereafter proliferated in the blood for at least 10 days before exiting the vascular system through capillary walls. From day 12 post-infection onwards, the kidney, as well as the spleen and the liver, were invaded. Numbers of S. truttae invading the kidney (37.3%) differed little from numbers found in the spleen (35.3%) and the liver (27.4%). The latter organs represented a dead end in the development of S. truttae as all stages in these organs degenerated and sporogony was found to take place exclusively inside the renal tubules. Early sporogonic stages were found from day 25 post-infection but mature spores only developed after at least 15 days of proliferation within the tubules.     

Organ and phospholipid class fatty acid specificity in response to dietary depletion of essential n‐3 fatty acids in Atlantic salmon (Salmo salar L.)

The cell membrane phospholipid composition is of major importance for normal cell functions. However, it is not known how complete depletion of both shorter and longer chain omega‐3 fatty acids in salmon diets influences fatty acid composition of phospholipid subclasses in different organs of Atlantic salmon. We describe here the fatty acid composition in phospholipid subclasses of liver, muscle, heart and intestine in Atlantic salmon after 18 months of dietary n‐3 essential fatty acids deprivation. The percentage of 22:6n‐3 was markedly reduced in almost all phospholipid subclasses, and except for muscle phosphatidylcholine, phosphatidylethanolamine (PE) and phosphatidylinositol (PI), phospholipids in deficient fish were totally devoid of 20:5n‐3. As compensation, we observed significant increases in 20:4n‐6, and especially in 20:3n‐9 (Mead acid) and 22:5n‐6, varying among phospholipids and organs. High amounts of 20:3n‐9 were found in liver and intestinal PE, little in PE from heart and muscle. For 22:5n‐6, we saw a small incorporation in PI in liver and intestine compared to heart and muscle. Generally in PI, the preference for 20:4n‐6 to 20:5n‐3 differed significantly between organs. Overall, changes upon n‐3 deprivation seemed to be strongest in liver and intestine, the lipid‐secreting organs, and less in muscle and heart.     

Florfenicol in Atlantic salmon, Salmo salar L., parr: tolerance and assessment of efficacy against furunculosis

Abstract. The efficacy of florfenicol against laboratory-induced infection with Aeromonas salmonicida was tested in Atlantic salmon, Salmo salar L., parr. Medication at three dose levels in the feed was started 24 h after bath challenge with A. salmonicida. The specific mortality rate in the unprotected infected control group was 75% compared with 5, 13 and 17% when florfenicol was given at dose levels of 20, 10 and 5 mg per kg body weight per day, respectively. Florfenicol was palatable to the fish at doses in excess of effective therapeutic levels and feeding for 10 days at 100 mg per kg, or for prolonged periods at 50 mg per kg and 10 mg per kg, resulted in no feeding problems or histopathological abnormalities. Florfenicol appears to have a good therapeutic index and considerable potential in the control of furunculosis in salmon.     

Effect of fish feed processing conditions on digestive protease activities, free amino acid pools, feed conversion efficiency and growth in Atlantic salmon (Salmo salar L.)

The responses of the digestive proteases trypsin and chymotrypsin and protein metabolism to differences in feed protein quality were investigated in Atlantic salmon (Salmo salar L.). Two sets of experimental feeds were produced. Each set of high and low quality feeds was provided to either 150 g or 2 kg salmon. Protein in the high quality feeds had significantly higher percentages of free (reactive) sulphydryl (SH) groups than the corresponding feeds based on low quality meals. After 90 days feeding, groups given high and low quality feeds did not differ in their specific growth rates (SGR) in either experiment. However, feed conversion efficiency (FCE) was significantly different between the high and low quality feed groups in 2 kg salmon, where the difference between the high and low feed protein qualities was larger, 10% versus 4% SH/[SH + (S–S)] in 150 g salmon. Higher FCE was preceded by significantly higher trypsin and chymotrypsin specific activities on day 60. SGR, in general, changed after the first month and was stable during the last 2 months in both experiments. Concurrently, both trypsin (T) and chymotrypsin (C) decreased with an increased activity ratio of trypsin to chymotrypsin (T/C ratio), and resulting in significantly lower T/C ratio on day 90 in salmon feeding on high quality feeds in both sizes of fish. Differences in FCE were associated with significant differences in levels of total free amino acids (TFAA) in the plasma and the white muscle, as well as in the ratio of essential to non‐essential free amino acids (EAA/NEAA ratio), free hydroxyproline, and RNA in the white muscle. Interestingly, after 3 days starvation (day 93), 5–7 h postprandial EAA/NEAA ratio in the plasma was significantly lower in the high quality diet groups in both experiments. Trypsin specific activity inversely correlated with muscle TFAA levels in 2 kg salmon, concurrent with higher muscle levels of RNA, lower free hydroxyproline and higher FCE in fish fed higher quality diets.     

Gene expression of fatty acid-binding proteins, fatty acid transport proteins (cd36 and FATP) and β-oxidation-related genes in Atlantic salmon (Salmo salar L.) fed fish oil or vegetable oil

Relative gene expression pattern of fatty acid transport proteins (FATP and cd36), intracellular fatty acid-binding proteins (FABP3, FABP10 and FABP11), β-oxidation-related genes [carnitine palmitoyl transferase II (CPTII), peroxisome proliferator-activated receptor β (PPARβ), acyl-CoA oxidase (AOX), long-chain fatty acyl-CoA synthetase (FACS), acyl-CoA dehydrogenase (dehydrogenase)] and uncoupling protein 2 (UCP2) was assessed by RT-qPCR in Atlantic salmon muscle (red and white), liver, heart, myosepta and visceral fat. FABP11, a FABP isoform not previously described in Atlantic salmon, was highly expressed in visceral fat and myosepta and at the lower level in red muscle, white muscle, myosepta and heart. Furthermore, Atlantic salmon were fed either a diet containing fish oil (FO) or a complete replacement of FO with a vegetable oil blend (55% rapeseed oil, 30% palm oil and 15% linseed oil; VO) for the production cycle (27 months from start of feeding and until ∼4.5 kg mean weight). The expression of genes related to β-oxidation, fatty acid uptake and transport in the white muscle indicate ( n  = 3) significant down-regulation in VO fed Atlantic salmon and correlated with previously reported white muscle triacylglycerol stores and β-oxidation. FABP11 in visceral fat and myosepta was also down-regulated in VO fed fish.     

Selective breeding can increase resistance of Atlantic salmon to furunculosis, infectious salmon anaemia and infectious pancreatic necrosis

We reasoned that by challenging large numbers of Atlantic salmon families with the causative agents of furunculosis, infectious salmon anaemia (ISA) and infectious pancreatic necrosis (IPN), we could show unequivocally that resistance to these diseases expresses moderate‐to‐high levels of additive genetic variation, and that the resistances are weakly correlated genetically. We tested this reasoning by challenging Atlantic salmon from 920 (approximately) full‐sib families with the causative agents of furunculosis and ISA, and fish from 265 of these families with the causative agent of IPN. Additive genetic variation and genetic correlations were estimated by fitting a threshold liability model to the resistances assessed as binary traits. Resistance to furunculosis, ISA and IPN was moderate –to highly heritable. The marginal posterior means for heritability on the underlying liability scale were 0.37 for resistance to ISA, and 0.55 and 0.62 for resistance to IPN and furunculosis. Genetic correlations between the resistances were weak (?0.11 to 0.07). These levels of additive genetic variation indicate that resistance to furunculosis, ISA and IPN will respond to selection. The weak genetic correlations indicate that it should be relatively easy to improve resistance to the diseases simultaneously. We believe that there is now strong evidence that selectively breeding Atlantic salmon for resistance can be highly successful.     

Induction of infectious pancreatic necrosis (IPN) in covertly infected Atlantic salmon, Salmo salar L., post-smolts by stress exposure, by injection of IPN virus (IPNV) and by cohabitation

Atlantic salmon post-smolts were given an intraperitoneal (ip) injection of tissue homogenate of Atlantic salmon fry from an outbreak of infectious pancreatic necrosis (IPN), and cohabitants were given an ip injection of Earle's balanced salt solution (EBSS). Parallel treatment groups were exposed to recurrent episodes of environmental stress by water drainage twice a week. Fish injected with EBSS and non-injected fish were exposed to water drainage. The control fish were left untreated. Mortality due to IPN started 3 weeks after challenge in non-injected and EBSS-injected fish that had been exposed to water drainage. This showed that the fish used in the experiment were covertly infected with IPN virus (IPNV) prior to challenge, although no virus was detected in the fish sampled before the experiment. In fish that received an injection of IPNV, mortality started 5-6 days after challenge, regardless of the presence or absence of stress exposure. The EBSS-injected cohabitants started to die after an additional 5-6 days, also regardless of the presence or absence of stress exposure. The final cumulative mortality in the IPNV-injected fish was significantly lower than in the EBSS-injected cohabitants, thus suggesting that the secondary immune response after injection of IPNV provided more protection than the response after a water-borne infection. No disease outbreak was observed in the control fish.     

Short-term storage and cryopreservation of milt from Atlantic salmon and sea trout

Experiments on short-term preservation of sperm were performed with Atlantic salmon (Salmo salar). Fertility was maintained for up to 10 days when 2 mm thick samples were stored at 0° C under an oxygen atmosphere in the presence of antibiotics (125 IU penicillin and 125 μg streptomycin per ml sperm). Fertility was completely lost after 24 days. Sperm stored without antibiotics fertilized 100% of eggs after 6 days.

Cryopreservation was carried out with milt from Atlantic salmon and sea trout (Salmo trutta). Semen mixed with extender was frozen on dry ice (pellets) with subsequent storage in liquid nitrogen. Sperm pellets were thawed in a 0.12-M NaHCO₃-solution (10° C) before insemination. The suitability of an extender as previously described by Stoss and Holtz and of a 0.3-M glucose solution with the addition of 10% DMSO, was tested on two different batches of sperm and eggs in Atlantic salmon and sea trout. In addition, the extender earlier reported by Mounib and an aqueous solution of 10% DMSO were only used in Atlantic salmon with one batch of gametes.

Insemination with cryopreserved Atlantic salmon sperm resulted in 36 to 91.3% eyed eggs (control = 100). The differences were caused by the type of extender and the batch of gametes employed. The very simple extender consisting of 0.3 M glucose and 10% DMSO only was the most successful. Results with cryopreserved sea trout sperm ranged between 38.6–54.8% eyed eggs, showing no difference between treatments.     

Anesthesia induces stress in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>), Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) and Atlantic halibut (<Emphasis Type="Italic">Hippoglossus hippoglossus</Emphasis>)

Stress in response to anesthesia with benzocaine, MS-222, metomidate and isoeugenol was studied in Atlantic salmon (Salmo salar), Atlantic halibut (Hippoglossus hippoglossus), and Atlantic cod (Gadus morhua) with no concomitant stress from handling or confinement in association with anesthesia or sampling. All of the anesthetics tested induced a stress response in all species, displayed by a release of cortisol to the water. MS-222 anesthesia elicited the highest cortisol release rates, reaching maximum levels 0.5 h post-exposure and returning to basal levels after 3–4 h. Benzocaine anesthesia caused a bimodal response where the initial peak in cortisol release rate was followed by a second increase lasting towards the end of the trial (6 h). This bimodality was more profound in Atlantic salmon than in Atlantic halibut and Atlantic cod. Metomidate anesthesia induced the lowest release of cortisol of the agents tested in both Atlantic halibut and Atlantic cod, but resulted in a bimodal response in Atlantic salmon where the initial increase in cortisol release was followed by a larger increase peaking at 2–2.5 h post exposure before returning to basal levels after 5 h. The stress induced in Atlantic salmon by isoeugenol anesthesia resembled that of MS-222, but did not reach the same elevated level. Overall, the cortisol release was most profound in Atlantic salmon followed by Atlantic halibut and Atlantic cod.     

Multiple-dose pharmacokinetic and depletion studies of sarafloxacin in Atlantic salmon, Salmo salar L.

Abstract. Two multiple-dose pharmacokinetic and depletion studies with sarafloxacin hydrochloride in feed pellets were conducted with Atlantic salmon at 12.1 ± 1.1 †C and 9.3 ± 1.5†C, respectively. The dose regimens used were 10mg sarafloxacin kg^-1 biomass for 10 days, and 20mg sarafloxacin kg^-1 biomass for 5 days. In the 10-day study, the highest average concentrations of sarafloxacin in plasma, muscle and liver were 0.14, 0.39 and 0.88μgg(ml)^-1, respectively. In the 5-day study, the highest average concentrations in plasma, muscle, liver and skin were 0.40, 0.61, 1.56 and 0.19μg(ml)^-1, respectively. A comparison between the individual plasma concentrations and the corresponding tissue concentrations revealed significantly higher concentrations in tissue than in plasma during the depletion phase. A similar comparison made in the therapeutic phase from 3 days after first medication to one day after the last medication revealed significantly higher concentrations in liver and muscle than in plasma, and significantly lower concentrations in skin than in plasma. On withdrawal of the drug, sarafloxacin concentrations in plasma and the different tissues declined rapidly, Sarafloxacin was not detected in any plasma sample taken 6 days or more after the end of medication. The corresponding figures for muscle, skin and liver tissues were 14, 20 and 22 days, respectively. The half-lives of sarafloxacin varied in the different tissues included in the studies. The half-life was shortest in plasma, and increased in ascending order in muscle, liver and skin.     

Effect of handling and fish size on secondary changes in carbohydrate metabolism in Atlantic salmon, Salmo salar L.

Two size groups (0.5 and 3 kg) of Atlantic salmon, Salmo salar L., were stressed by netting and transport from sea cages into land-based tanks. Handling lasted for 1 h. Before onset of stress, basal concentrations of haematocrit and haemoglobin, plasma cortisol and glucose, and liver and muscle glycogen were measured. All levels were within ranges reported for salmon. After handling and at chosen time intervals during recovery, samples were taken to judge the impact of handling and whether secondary changes in carbohydrate metabolism were related of fish size. A low cortisol peak indicated a mild stress reaction after handling independent to fish size. Plasma glucose peaked as cortisol declined, and returned to basal levels within 48 h. Liver glycogen seemed to be the main source of plasma glucose. No changes were measured in muscle glycogen concentrations. The results indicate a high tolerance to handling stress in Atlantic salmon independent of fish size.     

Feed intake and absorption of lipid oxidation products in Atlantic salmon (Salmo salar) fed diets coated with oxidised fish oil

Two questions were asked in the present study; does Atlantic salmon taste and discriminate against oxidised feed and are lipid oxidation products absorbed from the intestine. In Experiment 1, a control diet, a medium oxidised diet and a highly oxidised diet were prepared (TBARS levels: 34±5, 61±4 and 76±2 nmol g⁻¹, respectively). The control diet was marked with holmium and the experimental diets with europium. Each of the experimental diets were fed together with the control diet (1:1) to Atlantic salmon in duplicate tanks in one meal to satiation and the stomach contents were analysed for the lanthanides. The fish discriminated slightly against the highly oxidised diet, but not against the medium oxidised diet. In Experiment 2, Atlantic salmon cannulated through the dorsal aorta were fed control and oxidised feed (TBARS: 71±5 and 204±18 nmol g⁻¹) and blood samples were collected regularly over a nine days period. On day 9, the fish were sacrificed, and samples of muscle, liver, intestinal tissue and contents were taken and analysed for oxidation products, and vitamins C and E. Plasma TBARS increased 3-4 fold in response to the oxidised diet and there was a slight increase in muscle and liver TBARS. The data on peroxide value (PV) in the tissues showed large variation, but no differences between the groups were detected. PV and TBARS in the contents of the large intestine increased 7- and 1.6-fold, respectively, in response to oxidised feed. There were no differences in levels of vitamins C and E between the groups. It seems that Atlantic salmon feed quite well on oxidised feed and that aldehydes, here represented by MDA and measured as TBARS, are absorbed from the intestine. The question as to whether lipid hydroperoxides are absorbed is unanswered due to the large variation in tissue PV. On the other hand, animals in general seem to be protected from absorption of lipid hydroperoxides and we hypothesise that similar mechanisms are active in Atlantic salmon. This revised version was published online in August 2006 with corrections to the Cover Date.     

Growth and performance of Atlantic salmon, Salmo salar L., following administration of a rhabdovirus DNA vaccine alone or concurrently with an oil-adjuvanted, polyvalent vaccine

This research demonstrates for the first time an absence of growth-related side effects in Atlantic salmon, Salmo salar L., following the injection of a DNA vaccine alone or concurrently with a commercially available, polyvalent, oil-adjuvanted vaccine. Using weight and specific growth rate measurements, individually tagged Atlantic salmon were monitored for 2028 degree days (dd) post-vaccination. During this time, DNA-vaccinated fish did not differ in weight, length, condition factor or specific growth rate compared to unvaccinated control fish. While differences in weight were observed between unvaccinated control and concurrently vaccinated fish, there were no significant differences in weight, length, condition factor or specific growth rate between concurrently vaccinated fish and adjuvant-vaccinated fish, suggesting that only adjuvant vaccination affected growth. To further determine if concurrent injection of a DNA vaccine and a polyvalent, oil-adjuvanted vaccine had a physiological impact on the Atlantic salmon, swimming performance tests were performed at 106 dd post-vaccination with U _crit,1, U _crit,2, the U _crit recovery ratio (RR) and the normalized RR being similar to values obtained from unvaccinated control fish. In summary, this study shows that concurrent injection of a DNA vaccine and a polyvalent, oil-adjuvanted vaccine does not negatively influence the growth or swimming performance of Atlantic salmon compared to adjuvant vaccination alone.     

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.     

Saltwater performance of triploid Atlantic salmon Salmo salar L. brown trout Salmo trutta L. hybrids

Survival and growth in a saltwater net-pen of sexually immature triploid Atlantic salmon Salmo salar L. brown trout Salmo trutta L. hybrids was comparable to that of immature diploid Atlantic salmon. Following 17 months of freshwater rearing, the experimental fish were individually tagged and transferred to a saltwater net-pen where they were raised communally for 376 days. Initial and final average weights were 158 and 760 g per fish for the diploid Atlantic salmon and 209 and 1010 g per fish for the triploid hybrids; weights for the hybrids were significantly larger in both cases (α= 0.05). Survival from transfer to harvest was 43% for the Atlantic salmon and 48% for the hybrids. Average specific growth rate of fish which survived to harvest was 0.42% day^-1 for Atlantic salmon and 0.41% day^-1 for hybrids; these values were not significantly different. No significant differences were observed in average condition factor and dress-out percentage between crosses. Average gonadal weights and gonadosomatic indices were not significantly different for male diploid Atlantic salmon, female triploid hybrids and male triploid hybrids, but were significantly greater for female Atlantic salmon.     

Influence of dietary carbohydrate on blood chemistry, immunity and disease resistance in Atlantic salmon, Salmo salar L.

Abstract. The influence of dietary carbohydrate (CHO) on blood chemistry, immunity and disease resistance was studied in two experiments with Atlantic salmon, Salmo salar L. Moist diets with increasing amounts of digestible CHO ranging from 0 to 30% (dry weight) were used. In the first experiment with adult (0.5 kg) fish, blood haemoglobin concentration was negatively correlated with increasing dietary CHO level, while serum glucose and protein did not differ between the groups. Serum cortisol increased linearly in fish fed from 5 to 30% CHO. Serum haemolytic activity was negatively correlated with dietary levels of CHO. Humoral immune responses elicited after vaccination by intraperitoneal injection or by dip immersion with Vibrio salmonicida showed no differences according to diet 10 and 17 weeks post-vaccination. Mortality after challenge with live Aeromonas salmonicida by intraperitoneal injection was lowest in fish fed 10% CHO. In the second experiment with juvenile Atlantic salmon (3g), there were minor differences in body and organ weights. Plasma glucose, protein and cholesterol were elevated in fish fed the highest CHO levels. Fish exposed to immersion challenge with different water concentrations of Vibrio anguillarum showed no statistical differences in mortality. The studies indicate that varying dietary levels of CHO affected immunity and resistance to bacterial infections to a minor extent in Atlantic salmon at low water temperatures during freshwater and seawater stages.     

Influence of dietary blends of menhaden oil and canola oil on growth, muscle lipid composition, and thyroidal status of Atlantic salmon (Salmo salar) in sea water

The effects of various dietary blends of menhaden oil (MO) with canola oil (CO) on the growth performance, whole body proximate composition, flesh quality (muscle proximate and lipid composition) and thyroidal status of immature Atlantic salmon in sea water were studied.Atlantic salmon (initial weight, 145.2–181.3 g), held on a natural photoperiod and in 1100 L fibreglass tanks that were supplied with running, aerated (D.O., 9–10.5 p.p.m.), ambient temperature (8–10.5 °C) sea water (salinity, 28–30), were fed twice daily to satiation one of four isonitrogenous (36% digestible protein) and isoenergetic (18.8 MJ of digestible energy kg^-1) extruded high-energy diets for 112 days. All diets contained omega –3 (n-3) fatty acids in excess of requirements and differed only with respect to the source of the supplemental lipid which was either, 25% MO; 20.75% MO and 4.25% CO; 16.5% MO and 8.5% CO; or 12.25% MO and 12.75% CO. Thus, CO comprised, respectively, 0, 15.5, 31.2, or 47.0% of the total dietary lipid content (28% on an air-dry basis).Dissimilar percentages of saturated fatty acids in the dietary lipids were not found to be consistently related to the apparent gross energy digestibility coefficients of the diets. Atlantic salmon growth, dry feed intake, feed and protein utilization, percent survival, thyroidal status, and whole body and muscle proximate compositions were generally not influenced by the different sources of supplemental lipid. Therefore, our results suggest that canola oil may comprise as much as 47% of the lipid in high-energy grower diets for Atlantic salmon without compromising performance.The muscle lipid compositions generally mirrored those of the dietary lipids which, in turn, were influenced strongly by the concentrations and compositions of the CO and MO in the diet. Hence, as the dietary CO level was increased there were attendant increases in percentages of oleic acid (18:1(n-9)), linoleic acid (18:2(n-6)), total omega-6 (n-6) fatty acid content, and ratios of (n-6) to (n-3) and decreases of eicosapentaenoic acid (EPA; 20:5(n-3)), docosahexaenoic acid (DHA; 22:6(n-3)) and n-3 HUFAs (EPA & DHA) in the flesh lipids. The ranges for percentages of saturated and unsaturated fatty acids in the flesh lipids were, however, much less than those noted respectively in the dietary lipids probably because of selective metabolism of many of the former acids and some of the 18 carbon unsaturates for energy purposes.