An analysis of levels of infectious pancreatic necrosis virus in Atlantic salmon,Salmo salar L., broodstock in Scotland between 1990–2002

Throughout this study period the prevalence of infectious pancreatic necrosis virus (IPNV) in Scottish farmed Atlantic salmon was high in the marine environment but relatively low in fresh water. In order to minimize the risk of vertical transmission of infection from parent to progeny, all IPNV infected broodstock populations had to undergo testing of all fish for the virus at the time of stripping and eggs from positive parents were destroyed. Between 1990 and 2002 over 68 000 Atlantic salmon broodfish were individually screened for IPNV by cell culture isolation and enzyme linked immunosorbent assay. Generalized linear mixed models were used to assess the influence of geographical region, age, sex and year on IPNV prevalence in Atlantic salmon broodstock. This analysis determined that the age and sex of the broodfish and the geographical region of the broodstock stripping site did not have a statistically significant influence on IPNV prevalence within the broodstock parental population. However, there was a statistically significant temporal increase in IPNV prevalence from 1990 to 2002.     

Randomized clinical field trial of a bacterial kidney disease vaccine in Atlantic salmon,Salmo salar L.

A randomized, blinded clinical trial was performed to assess the relative effectiveness of five commercial and one experimental vaccine in a population of farmed fish experiencing a bacterial kidney disease (BKD) outbreak that occurred in one study cage that was part of a larger clinical field trial. A total of 6000 uniquely identified Atlantic salmon S1 presmolts were randomly assigned to vaccine groups in the hatchery and transferred to a commercial marine aquaculture site. Repeated sampling events to evaluate growth, inherent physical conditions and health status were carried out over the entire production cycle. During the second summer at sea, the study cage developed an outbreak of BKD that lasted approximately 240 days. The effectiveness of the selected vaccines was evaluated using survival analysis methods. The sole vaccine group offering protection for BKD was found to significantly decrease the hazard of dying (hazard ratio, HR = 0.68, P = 0.018) during the outbreak, compared to the industry standard, vaccine group. Additionally, during the outbreak, fish with a shortened operculum had a significantly decreased hazard (HR = 0.38, P = 0.033) compared to those fish with a normal operculum, while fish with jaw deformities had a significantly increased hazard (HR = 2.55, P = 0.001) compared to fish with normal jaw status.     

Retrospective analysis of Atlantic salmon (Salmo salar) marine growth and condition in the northwest Atlantic based on tag‐recovery data

The life history of North American Atlantic salmon (Salmo salar) is characterized by extensive round‐trip migrations between freshwater rearing habitats and marine feeding grounds off the coasts of Canada and Greenland. Growth is rapid during the marine migration, and growth rate and condition factor may be indicators of salmon health during this period. Growth data were evaluated from a tag‐recovery program conducted from 1969 to 1991 using hatchery‐reared Atlantic salmon smolts released in the Penobscot River, Maine, U.S.A. Information from recaptures of 3167 salmon that were at large in the marine environment for 1 month to 3 yr was analyzed. Length–weight measurements coupled with time‐at‐large data were used to estimate von Bertalanffy and allometric growth parameters specific to the marine phase. Variations in growth and condition factor in relation to smolt age, release date, and temperature conditions in the northwest Atlantic were also examined. The von Bertalanffy k parameter declined with ordinal release date, indicating faster growth rates during the first year of smolts released earlier in the spring. The 2‐yr‐old smolts had a larger k than 1‐yr‐old smolts, although 1‐yr‐old smolts grew to a larger asymptotic size. Sea surface temperature had variable effects on growth parameters and condition factor, with temperature at the beginning of the migration and in overwintering habitat during the first year at sea having the greatest influence on length–weight relationships. Determining the mechanisms that influence growth of individuals during the marine phase will help elucidate the factors responsible for historic growth trends, establishing a baseline for current research.     

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.     

Viral co‐infections in farmed Atlantic salmon,Salmo salar L., displaying myocarditis

Several different viruses have been associated with myocarditis‐related diseases in the Atlantic salmon aquaculture industry. In this study, we investigated the presence of PMCV, SAV, PRV and the recently identified Atlantic salmon calicivirus (ASCV), alone and as co‐infections in farmed Atlantic salmon displaying myocarditis. The analyses were performed at the individual level and comprised qPCR and histopathological examination of 397 salmon from 25 farms along the Norwegian coast. The samples were collected in 2009 and 2010, 5–22 months post‐sea transfer. The study documented multiple causes of myocarditis and revealed co‐infections including individual fish infected with all four viruses. There was an overall correlation between lesions characteristic of CMS and PD and the presence of PMCV and SAV, respectively. Although PRV was ubiquitously present, high viral loads were with a few exceptions, correlated with lesions characteristic of HSMI. ASCV did not seem to have any impact on myocardial infection by PMCV, SAV or PRV. qPCR indicated a negative correlation between PMCV and SAV viral loads. Co‐infections result in mixed and atypical pathological changes which pose a challenge for disease diagnostic work.     

Hypervitaminosis A in first‐feeding fry of the Atlantic salmon (Salmo salar L.)

Atlantic salmon fry were reared on a fishmeal based diet with increasing levels of vitamin A (VA) (6, 122 and 938 mg retinol kg^–1 dry feed) from startfeeding and for 14 weeks. Signs of VA stress, such as reduced fat stores, liver size and growth, were found for groups receiving 122 and 938 mg retinol kg^–1. Signs of vitamin A toxicity, such as increased mortality, abnormal vertebral growth, and reduced growth, were found for groups receiving 938 mg retinol kg^–1. These results suggest that excess VA in the early life stages of Atlantic salmon is deleterious for normal development.     

The impact of Aeromonas salmonicida infection on behaviour and physiology of Atlantic salmon (Salmo salar L.)

This study examined the effect of Aeromonas salmonicida infection on the swimming behaviour and physiology of Atlantic salmon (Salmo salar L.). Fish were injected in the dorsal muscle with either 100 μL bacterium solution (3.05 × 10⁷CFU mL^?1) Aeromonas salmonicida, or 100 μL 0.9% NaCl (as control group). Compared with the control group, the pathogen injected group significantly impaired the critical swimming speed (U_crit) and exhausting time (P < 0.05), which were reduced by 37% and 39% at severe time (day 6, when most of the fish challenged by Aeromonas salmonicida died) respectively. Furthermore, the blood parameters related to their swimming behaviour were also influenced significantly by pathogen injection (P < 0.05). The results showed that the high density lipoprotein (HDL), haemoglobin and total protein decreased by about 63%, 49% and 74% at the end, respectively, while lactate increased by about 29% on day 6. The results suggested that the swimming performance of Atlantic salmon might be a useful indicator of disease, and it was feasible to warn the outbreaks of acute disease by fish behaviour.     

Epitheliocystis in Atlantic salmon,Salmo salar L., farmed in fresh water in Ireland is associated with ‘Candidatus Clavochlamydia salmonicola’ infection

Intracellular inclusions containing chlamydia‐like organisms are frequently observed in the gill epithelial cells of Atlantic salmon, Salmo salar L., cultured in fresh water in Ireland. In this study, the causative agent was identified in four separate freshwater sites, using 16s rRNA sequencing, as ‘Candidatus Clavochlamydia salmonicola’. Histopathology and real‐time (RT) PCR were used to further assess infections. The prevalence of infection ranged from 75–100% between sites and infection intensity was highly variable. No significant lesions were associated with these infections. As a diagnostic tool, RT‐PCR proved marginally more sensitive than histopathology. The fate of ‘Candidatus Clavochlamydia salmonicola’ in Atlantic salmon post‐seawater transfer was investigated in a 12‐week marine longitudinal study. Both RT‐PCR and histopathological examination indicate that the organism disappears from the gills 4–6 weeks post‐transfer.     

Rapid compositional analysis of Atlantic salmon (Salmo salar) using visible‐near infrared reflectance spectroscopy

Rapid measurement of salmon flesh quality parameters (>400 samples day^?1) was demonstrated in the laboratory and remotely at industrial sites. Visible‐near infrared spectroscopy (VNIRS) was applied to predict astaxanthin (AX) and fat content in farmed Atlantic salmon. Fish were sampled from thirteen batches (1–6 kg whole weight, containing 2.3–16.3% fat and 1.2–12.5 μg g^?1 AX), and models validated on small (average ± SD: 1.4 ± 0.4 kg) and large fish (4.2 ± 0.9 kg). Both constituents were well predicted in minced Norwegian Quality Cutlet (NQC) samples (r² ≥ 0.86; standard error of prediction (SEP) ≤0.7% for fat and ≤0.7 μg g^?1 for AX). Comparable metrics were observed for AX prediction in whole NQCs (r² = 0.80–0.88; SEP 0.7 μg g^?1). Fat was better predicted in small fish than large fish for whole NQCs (r² = 0.82, SEP 1.0% cf r² = 0.59, SEP = 0.59%) and non‐destructive scanning through the skin of whole, gutted fish (r² = 0.77, SEP = 1.2% cf r² = 0.49, SEP = 1.5%). Models were also developed for screening polyunsaturated fatty acid (PUFA) concentrations, e.g. in NQCs for docosahexaenoic acid (r² = 0.73) and n‐3:n‐6 PUFA (r² = 0.89).     

Piscine reovirus (PRV) in wild Atlantic salmon,Salmo salar L., and sea‐trout,Salmo trutta L., in Norway

This is the first comprehensive study on the occurrence and distribution of piscine reovirus (PRV) in Atlantic salmon, Salmo salar L., caught in Norwegian rivers. PRV is a newly discovered reovirus associated with heart and skeletal muscle inflammation (HSMI), a serious and commercially important disease affecting farmed Atlantic salmon in Norway. A cross‐sectional survey based on real‐time RT‐PCR screening of head kidney samples from wild, cultivated and escaped farmed Atlantic salmon caught from 2007 to 2009 in Norwegian rivers has been conducted. In addition, anadromous trout (sea‐trout), Salmo trutta L., caught from 2007 to 2010, and anadromous Arctic char, Salvelinus alpinus (L.), caught from 2007 to 2009, were tested. PRV was detected in Atlantic salmon from all counties included in the study and in 31 of 36 examined rivers. PRV was also detected in sea‐trout but not in anadromous Arctic char. In this study, the mean proportion of PRV positives was 13.4% in wild Atlantic salmon, 24.0% in salmon released for stock enhancement purposes and 55.2% in escaped farmed salmon. Histopathological examination of hearts from 21 PRV‐positive wild and one cultivated salmon (Ct values ranging from 17.0 to 39.8) revealed no HSMI‐related lesions. Thus, it seems that PRV is widespread in Atlantic salmon returning to Norwegian rivers, and that the virus can be present in high titres without causing lesions traditionally associated with HSMI.     

Transport of alpha-tocopherol in Atlantic salmon (Salmo salar) during vitellogenesis

The transport of α-tocopherol was studied during vitellogenesis in Atlantic salmon that were fed diets with two levels of α-tocopherol. α-Tocopherol levels were measured in the flesh, liver, ovary and serum, and in the serum the α-tocopherol levels in the very low density lipoprotein (VLDL), low density lipoprotein (LDL), high density lipoprotein (HDL) and very high density lipoprotein (VHDL or vitellogenin) were also measured. Atlantic salmon store α-tocopherol mainly in their flesh because the muscle mass comprises 50% or more of live weight. During vitellogenesis the α-tocopherol content declined to about 10% of the level prior to maturation. The relative range of level of α-tocopherol in the lipoproteins was: HDL> LDL> VLDL> VHDL, irrespective of dietary levels of α-tocopherol. From the recent knowledge on lipid transport during vitellogenesis and the present data, we hypothesize that α-tocopherol is transported from peripheral tissues to liver by HDL and further transported from liver to ovary by LDL. Vitellogenin appears to play a minor role in the transportation of vitamin E to the ovary.     

Stable isotope profiling for large‐scale evaluation of feed efficiency in Atlantic salmon (Salmo salar)

In growing animals, individual variation in feed efficiency may arise from individual differences in growth rate and protein metabolism. Over a period of time, these factors will affect the ratio between ‘new’ vs. ‘old’ protein, which can be quantified using isotope profiling. The aim of this study was to investigate the relationship between relative weight gain and atom percentage excess ¹⁵N in the muscle, liver and mid‐intestine. A 50‐day experiment was conducted with a total of 375 fish initially fed a standard diet, subsequently replaced by one out of five experimental diets, enriched with ¹⁵N. In general, fast‐growing fish are expected to have a better feed efficiency, and the results show that this is captured by isotope profiling in liver and muscle tissues. Furthermore, individual variation in isotope content, that is relative fraction of ‘new’ protein, among fish with comparable growth rates was observed, most expressed around ~50% isotope saturation, indicating differences in protein degradation and replacement not attributed to growth. The results suggest that isotope profiles can be used as individual indicator traits for feed efficiency and that inclusion levels of stable isotopes of 1%–2% gave the most reliable results.     

An outbreak of acute disease and mortality in Atlantic salmon (Salmo salar) post‐smolts in Norway caused by Tenacibaculum dicentrarchi

An outbreak of disease characterized by skin ulcers, fin rot and mortality was observed a few days after the transfer of Atlantic salmon (Salmo salar) from a freshwater smolt production facility to a land‐based seawater post‐smolt site. Dead and moribund fish had severe skin and muscle ulcers, often 2–6 cm wide, particularly caudal to the pectoral fins. Microscopic examination of smears from ulcers and head kidney identified long, slender Gram‐negative rods. Histopathological analysis revealed abundance of long, slender Tenacibaculum‐like bacteria in ulcers and affected fins. Genetic characterization using multilocus sequence analysis (MLSA) of seven housekeeping genes, including atpA, dnaK, glyA, gyrB, infB, rlmN and tgt, revealed that the isolates obtained during the outbreak were all clustered with the Tenacibaculum dicentrarchi‐type strain (USC39/09^T) from Spain. Two bath challenge experiments with Atlantic salmon and an isolate of T. dicentrarchi from the outbreak were performed. No disease or mortality was observed in the first trial. In the second trial with a higher challenge dose of T. dicentrarchi and longer challenge time, we got 100% mortality within 48 hr. This is the first reported outbreak of disease caused by T. dicentrarchi in Norwegian farmed Atlantic salmon.     

Adult Atlantic salmon (Salmo salar L.) adapts to long‐term surplus dietary arginine supplementation

The current study aimed to investigate the effect of surplus dietary arginine on polyamine and amino acid metabolism and accretion of proteins and lipids, as previous studies have demonstrated a lipid‐reducing effect following surplus arginine supplementation in viscera as well as increased muscle growth. Four plant protein‐based diets were given increasing concentrations of L‐arginine, from 21.1 to 36.1 g kg^‐1 DM, and fed to quadruplicate tanks of adult Atlantic salmon (1.1 kg) for 12 weeks. No effects on growth or deposition of fat or protein were observed. Free amino acid concentrations and related metabolites were unaffected in the liver, except for urea, while concentrations in muscle and plasma reflected production of arginine metabolites. Polyamine concentrations were unaffected in liver, muscle and white adipose tissue (WAT), as were the abundance and activity of spermidine/spermine N1‐acetyltransferase (SSAT), the rate‐limiting enzyme in polyamine turnover. Gene expression demonstrated differential regulation of ornithine decarboxylase (ODC) in liver and WAT, although overall little effects were observed on gene expression. Liver S‐adenosylmethionine (SAM) concentrations decreased with arginine supplementation. We suggest that adult Atlantic salmon have adapted to surplus arginine, and the main long‐term effect appears to be increased concentrations of arginine metabolites.     

An experimental investigation on aspects of infectious salmon anaemia virus (ISAV) infection dynamics in seawater Atlantic salmon, Salmo salar L.

This study investigated infection dynamics of infectious salmon anaemia virus (ISAV) by conducting two experiments to examine minimum infective dose and viral shedding of ISAV. In terms of minimum infective dose, the high variability between replicate tanks and the relatively slow spread of infection through the population at 1 × 10¹ TCID₅₀ mL⁻¹ indicated this dose is approaching the minimum infective dose for ISAV in seawater salmon populations. A novel qPCR assay incorporating an influenza virus control standard with each seawater sample was developed that enabled the quantity of ISAV shed from infected populations to be estimated in values equivalent to viral titres. Viral shedding was first detected at 7 days post-challenge (5.8 × 10⁻² TCID₅₀ mL⁻¹ kg⁻¹) and rose to levels above the minimum infective dose (4.2 × 10¹ TCID₅₀ mL⁻¹ kg⁻¹) on day 11 post-challenge, 2 days before mortalities in ISAV inoculated fish started. These results clearly demonstrate that a large viral shedding event occurs before death. Viral titres peaked at 7.0 × 10¹ TCID₅₀ mL⁻¹ kg⁻¹ 15 days post-infection. These data provide important information relevant to the management of ISA.