Duration and method of fixation affects the sensitivity of a digoxygenin-labelled DNA probe in detecting Kudoa thyrsites in Atlantic salmon skeletal muscle

The effect of fixative and duration of fixation on the sensitivity of a non-radioactive in situ hybridisation (ISH) protocol to detect Kudoa thyrsites small subunit ribosomal deoxyribonucleic acid (DNA) was investigated. Strong ISH reactions were detected in 5-μm sections of paraffin-embedded Atlantic salmon muscle after fixation for 1 day in Davidson's solution (DS). Reactions were weak following 3 or 5 days fixation and absent after 17 or 28 days fixation. Strong ISH reactions were observed after 1, 3 or 5 days fixation in neutral buffered formalin (NBF). The reactions were weak after 17 days and weak to nonexistent after 28 days of fixation. Reactions were consistently strong after fixation in 95% ethanol for up to 28 days. Some mature spores reacted weakly or not at all by ISH. Parasite DNA was weakly amplified by polymerase chain reaction (PCR) from paraffin-embedded muscle after 1 day of fixation in DS but not after fixation for 3, 5, 17 or 28 days. Amplified DNA was detected after fixation in NBF for 1, 3 and 5 days, but not after 17 or 28 days. In contrast, PCR consistently amplified DNA from paraffin-embedded, ethanol-fixed muscle. Caution should be used in the choice of fixative and duration of fixation when preserving Atlantic salmon tissues for molecular diagnosis of K. thyrsites.     

Is there lipostatic regulation of feed intake in Atlantic salmon Salmo salar L.?

High‐energy feeds, with increased fat concentrations, are often used in salmon farming to improve feed:gain ratio. However, fish are thought to regulate ingestion to meet their energy and nutrient intake requirements. Further, feeds with excessive fat content will lead to increased adiposity, which is thought to exert a negative feedback on feed intake via lipostatic regulation mechanisms. A test of the lipostatic model of feed intake regulation was carried out on juvenile Atlantic salmon Salmo salar L. (c. 165 g) in which body fat content (5.6% and 9.4% body fat) had been manipulated by feeding feeds with different fat concentrations. Thereafter, the fish were offered the high‐ and low‐fat feeds (15.6% and 26.5% dietary fat content) simultaneously, and our hypothesis was that feed preference and intake would reflect the state of fat storage. Thus, we predicted that, when given a choice of feeds, the lean fish would eat more, and that the body fat status of the groups would converge over time. The results indicated a general preference for the leaner feed irrespective of adiposity level, but leaner fish consumed more feed, grew faster and deposited more body fat than their fatter counterparts. Over time, body compositions converged among treatments, and differences in feed intake ablated. These findings seem to provide supportive evidence for a lipostatic regulation of feed intake in fish.     

Dietary inclusion of crystalline D- and L-methionine: effects on growth, feed and protein utilization, and digestibility in small and large Atlantic salmon (Salmon salar L.)

Use of crystalline methionine to optimize amino acid profiles in feeds for Atlantic salmon (Salmo salar L.) was studied in three experiments. A basic experimental diet with 25% of dietary protein from marine origin and the remaining 75% from soya protein concentrate was used either with or without supplementation of L ‐methionine, D ‐methionine or DL ‐methionine. A fishmeal‐based diet was used as control. In experiment 1, growth performance of 100 g salmon in seawater was improved by supplementation with the different methionine products. Growth performance was, however, inferior to that of fish fed the control. Feed conversion ratios (FCR) exhibited similar trends to specific growth rate (SGR) with methionine supplementation, but again not as the control. Protein retention, measured as productive protein value (PPV), was significantly better in the control and D ‐methionine groups as compared with either L ‐methionine or the nonsupplemented group. These trends were confirmed in experiment 2, using fish of the same size, where an absorption and digestibility study of D ‐ and L ‐methionine clearly indicated that D ‐methionine was absorbed as well as L ‐methionine. Results indicated that racemization of D ‐ into L ‐methionine occurs in liver and kidney in Atlantic salmon. Ornithine decarboxylase (ODC) activity in liver was significantly affected by dietary treatment and there was a good correlation with growth and feed utilization data. In experiment 3, a growth study with large salmon (1 kg) was performed using increasing supplementation of DL ‐methionine to the basic diet. Results indicated a methionine requirement for maximum growth of 9.2 g kg^–1 feed dry matter (DM) kg^–1 growth. This is slightly higher than that reported in the literature. The present study clearly shows that optimizing the amino acid profile by inclusion of a single crystalline amino acid (crystalline methionine) does not give the same protein utilization as that of a diet in which the amino acids are mainly protein bound. D ‐methionine was found to result in better protein utilization than L ‐methionine when measured as PPV.     

Changes in density of under-yearling masu salmon <Emphasis Type="Italic">Oncorhynchus masou</Emphasis> in a natural stream after release

ABSTRACT:   The relation between the density of fish and parameters such as body weight, condition factor and the hepatic triglyceride (TG) content of masu salmon Oncorhynchus masou juveniles after release into a natural stream were studied. Hatchery-reared juveniles of different sizes and numbers were released in May 1999, 2000 and 2001, and investigations were carried out from July to September each year. The density of fish was relatively high in 1999, when juveniles of the largest size and number were released, but then continued to decrease during the investigation. The hepatic TG content remained considerably low in 1999, and there was a significant negative correlation between the density of fish and the hepatic TG content of the juveniles caught over three years. The initial fish size and number at release as well as the nutritional condition of the juveniles may affect the density of fish after release.     

Changes in rhodopsin-porphyropsin ratio of chum and pink salmon

ABSTRACT:   In order to investigate the changes in rhodopsin-porphyropsin ratio of chum salmon and pink salmon in relation to the change in their habitat, the ratios were analyzed in individual fish prior to stocking, during the sea run, homing, and upstream migration. The ratio in both the species gradually increased during the sea run. However, the ratio decreased after upstream river migration. Moreover, in the sea, the rhodopsin-porphyropsin ratio of chum salmon was always slightly higher than that of pink salmon. The largest difference in the individual variation was observed in the individuals caught with a set net placed near the mouth of a home river.     

Growth and feed utilization of large size rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) reared in freshwater: diet and species effects, and responses over time

Four diets differing in crude protein/crude lipid concentrations (CP/CL), 570/200, 510/220, 460/240, 430/260 (g kg^?1 dry diet) were fed to near‐satiety to rainbow trout (initial body weight, IBW = 268 g, domestic strain) and Atlantic salmon (IBW = 456 g, wild strain) for 308 days to determine the effect of diets, species and fish size on efficiency of feed, nitrogen (N) and energy utilization. Within species, weight gain, feed efficiency (FE), and energy retention efficiency (ERE, E gain/E intake) were not affected by diet (P < 0.05). N retention efficiency (NRE, N gain/N intake) increased linearly (P < 0.05) with decreasing CP/CL for both species. NRE and FE were higher for salmon compared with trout. Whole body CP of salmon was significantly (P < 0.05) higher, and lipid significantly (P < 0.05) lower, than those of trout. There was a significant (P < 0.05) linear decrease in FE as fish grew, regardless of diet and species. NRE linearly decreased (P < 0.0001) and lipid to protein deposition ratio (LD/PD) increased (P < 0.05) as trout grew. NRE linearly increased (P < 0.05) while ERE and LD/PD linearly decreased (P < 0.05) as salmon grew. Salmon appeared to rely to a greater extent on non‐protein energy‐yielding nutrients to support cost of growth than did trout.     

Purification of chinook salmon (Oncorhynchus tshawytscha) GH for receptor study

A method for the purification of chinook Salmon (Oncorhynchus tshawytscha) GH, which retains its biological activity, is described. The biological activity was investigated with an established radioreceptor assay using liver membranes from pregnant rabbits and bovine GH as standard and labelled hormone. The enrichment of the preparation was checked with electrophoresis (SDS-PAGE). Extraction and further steps were carried out using low molarity alkaline buffer (pH 8–10, M = 100 mM). Three chromatography steps were performed (Concanavalin-A sepharose, Bio-gel P60, DEAE). Ion exchange chromatography was performed under isocratic conditions (using a 50 cm column). Two isoforms (sGH1 and sGH2) were isolated. The purification yield is 0.7% compared to lyophilized pituitaries. The molecule is homogeneous in SDS-PAGE. Contamination by prolactin, gonadotrophin and corticotrophin is negligible (< 0.5%). It could be demonstrated that the biological activity of the preparation is maintained since this preparation stimulates the growth of juvenile trout (Salmo gairdneri) and binds specifically (35%) to trout liver membranes.     

Heart and skeletal muscle inflammation in Atlantic salmon, Salmo salar L: a new infectious disease

Heart and skeletal muscle inflammation (HSMI) is a disease syndrome of unknown aetiology first observed in farmed Atlantic salmon, Salmo salar, in 1999. In the present study we have demonstrated for the first time that HSMI is an infectious disease. It was induced in Atlantic salmon post-smolts after injection with tissue homogenate from farmed Atlantic salmon previously diagnosed with HSMI. The lesions were also induced in cohabitating salmon given a corresponding injection without tissue homogenate. Six weeks post-challenge the fish that had been injected with tissue homogenate developed a serious epicarditis and myocarditis with mononuclear cell infiltrations in compact and spongy layers of the heart. Similar lesions were found in cohabitants after 10 weeks. The lesions were consistent with samples from field outbreaks of HSMI. No lesions were found in control fish. A viral aetiology is strongly suggested, as no difference in disease induction between an inoculum containing antibiotics and a non-treated inoculum was found. Further investigations are required in order to make conclusions regarding the cause and pathogenesis of HSMI.