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.     

Studies with experimental transmission of heart and skeletal muscle inflammation in Atlantic salmon, Salmo salar L.

Heart and skeletal muscle inflammation (HSMI) is a transmissible disease causing mortality in farmed Atlantic salmon, Salmo salar L. It is characterized by epi-, endo- and myocarditis and myocardial necrosis, as well as myositis and necrosis of red skeletal muscle. The present paper describes two infection experiments, with the aim of further exploring the infectivity and pathogenesis of HSMI. In both experiments, Atlantic salmon were intraperitonally injected with putatively infective material. The first experiment was carried out in fresh water, using cardiac tissue, blood plasma and cell cultured material as inoculates. In the second experiment, various tissues sampled from fish in the mid-outbreak phase were used to inoculate experimental fish in sea water. Also, cardiac tissue sampled before and after the outbreak phase was used. Finally, cardiac tissue pre-treated with chloroform was tested. In both experiments, all inoculates resulted in cardiac inflammation during the study period of 8 weeks. Early cardiac changes included perivasculitis and endocarditis, which were observed from 1–3 weeks post-challenge (p.c.). Focal myocarditis first appeared 3 weeks p.c., and the number of fish showing myocardial changes at 8 weeks p.c. was high in all groups. A possible mechanism for the development of HSMI is discussed.     

Plasma protein changes in Atlantic salmon, Salmo salar L., with infectious salmon anaemia

Moderate to severe anaemia and hypoproteinaemia were reported in a Canadian outbreak of 'haemorrhagic kidney syndrome' in Atlantic salmon, later shown to be caused by a variant of infectious salmon anaemia virus (ISAV). The progressive anaemia associated with ISA has been previously reported, but hypoproteinaemia in salmon infected with European isolates of ISA virus has not been well documented. The present study showed a very significant positive correlation between decreasing haematocrit values and total plasma protein concentrations in Atlantic salmon infected with two Canadian and two Norwegian ISA viral isolates. However, variations in the concentration of individual plasma proteins, typical of acute phase responses in higher vertebrates, were not observed.     

Longitudinal study of a natural outbreak of heart and skeletal muscle inflammation in Atlantic salmon, Salmo salar L

Heart and skeletal muscle inflammation (HSMI) is a transmissible disease of farmed Atlantic salmon, Salmo salar L. It is characterized by significant epi-, endo- and myocarditis, as well as myositis, particularly involving red skeletal muscle. The aetiology of HSMI is currently unresolved, though a viral cause is suspected. Since its discovery in 1999, HSMI has become an increasing problem for the Norwegian farming industry, with some farms experiencing yearly outbreaks and subsequent economic losses. In the present study an Atlantic salmon farm was studied from December 2003 to April 2005. Samples from apparently healthy as well as clinically diseased fish were collected monthly and examined histopathologically. The first fish to be diagnosed with HSMI was sampled in May, 8 months after transfer to sea. A clinical outbreak of HSMI followed in June, when all fish in the sample had lesions consistent with HSMI. Subsequent samples revealed that cardiac lesions decreased in severity 2 months after the start of the outbreak, but that multiple foci of cellular infiltration and necrosis persisted throughout the year. There appeared to be a shift in lesion location from being most severe in the compact myocardium in early stages of disease to a greater involvement of the atrium and spongy layer of the ventricle in later samples. Late samples also showed increased fibrosis of cardiac tissue. In conclusion, HSMI appears to be a severe disease with elevated mortality, morbidity close to 100% and prolonged duration.     

Screamer disease in Atlantic salmon, Salmo salar L., in Chile

Investigation of cranial and spinal deformities in Atlantic salmon smolts from Chile immediately after sea transfer, over two successive crops, demonstrated ankylosis of the mandibular articulation, spinal foreshortening, fracture of vertebrae and rarefaction of osseous and cartilaginous tissues including the operculum. As a result the mouth was permanently agape and opercula could be folded back on themselves. All affected fish had been transferred to sea water at 20 °C plus and fed on particular commercial diets. Fish in cooler areas, or on diets high in vitamin C and phosphorus in the high temperature zone, were clinically normal. The condition has not recurred following dietary adjustment.     

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.     

Influence of dietary lipid composition on cardiac pathology in farmed Atlantic salmon, Salmo salar L

The present study investigated the short-term (5 months) effect of replacing dietary marine oils with vegetable oils on the development of arteriosclerotic changes in the heart of Atlantic salmon, Salmo salar. The experiment was performed as a randomized observer-blinded and controlled trial. Farmed Atlantic salmon were randomly sampled from a study population containing 900 individuals. The salmon were divided into three groups and given diets with either 100% fish oil (Diet 1), a 50/50% mixture of fish oil and rapeseed oil (Diet 2) or 100% rapeseed oil (Diet 3). Ten sexually immature salmon from each dietary group were sampled in March and August 2002. Additionally, 47 sexually mature wild salmon were randomly collected in mid-September 2001. Serial histological sections were taken from the bulbus arteriosus and ventricle wall for histopathological evaluation of the coronary arteries and myocardium. No significant differences in mean coronary changes recorded by the main variable 'mean range lesion' (MRL) were detected between the groups in March or August. MRL increased significantly between March and August with Diet 2 (P < 0.01), was nearly significant with Diet 3 (P = 0.06) and was unchanged with Diet 1. This pattern coincided with the Diet 2 group having the highest increase in heart weight. MHC class II immunoreactive cells in the coronary changes were detected in sections from one individual in each group. Heart weight was the most dominant variable in the data set and explained linearly 15.5% of the variation in MRL. Body weight, fish length and heart weight were all significantly, positively and linearly correlated to MRL. The Diet 2 group had the highest growth rate and also exhibited a significant increase in MRL. The possible influence of diet composition on weight gain and MRL needs to be further elucidated. Increase in heart weight seems to be the dominating predictor of the appearance of MRL in Atlantic salmon. However, the present results cannot exclude the possibility that differences in fatty acid composition of fish feed can influence the development of arteriosclerotic changes in Atlantic salmon.     

Association of spinal deformity and vaccine-induced abdominal lesions in harvest-sized Atlantic salmon, Salmo salar L

Spinal deformities in Atlantic salmon, Salmo salar L., have been described as a disease of multifactorial origin for which vaccines and time of vaccination have been suggested as risk factors. A vaccine efficacy trial where spinal deformity became evident was continued by the observational study reported here. In the preharvest part of the study 17 months post-sea transfer, there was a prevalence of 11.3% spinal deformity, with deformities present only in one vaccine group indicating a strong vaccine involvement. At slaughter, the prevalence of spinal deformities was 11.7%, and deformed fish had only 62% of normal slaughter weight. Visual analogue scales (VAS) were used for continuous recordings of vaccine-induced abdominal lesions and deformity. A logistic regression model associating presence of spinal deformity with markers of abdominal lesions was developed. The odds ratio for spinal deformity was 5.7 (95% CI: 3.4–9.4) for each unit increase in adhesion score (0–6) and 4.9 (2.9–3.4) for each unit increase in melanin on abdominal organs (0–3). Lesions in the dorsal caudal part of the abdomen gave an odds ratio for spinal deformity of 2.2.     

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.     

Dietary phosphorus requirement of juvenile Atlantic salmon, Salmo salar L.

The objective of this study was to determine the dietary phosphorus (P) requirement of juvenile Atlantic salmon, Salmon salar L. Triplicate groups of fish (mean initial weight 1.4 g) were fed semipurified, casein-gelatine-based diets containing one of five levels of P (4, 8, 10, 15 and 25 g kg⁻¹) from Ca(H₂PO₄)₂·H₂O, or a commercial feed (17 g kg⁻¹ P) for 9 weeks. Weight gains did not differ significantly among treatment groups fed the experimental diets but were slightly less than gains in fish fed the commercial feed. Feed efficiency (wet weight gain/dry feed consumed) was similar in all groups, averaging 1.45. Availability of dietary P, estimated from apparent retention and apparent digestibility, was 86%. Whole-body P concentrations declined in fish fed diets containing less than 10 g kg⁻¹ P. Fitting a logistic curve to dietary P vs. whole-body P concentrations indicated that a minimum of 11 g kg⁻¹ dietary P (9 g kg⁻¹ digestible P) was required by juvenile Atlantic salmon to maintain whole-body P concentrations at initial levels. Calculation of a dietary requirement using a simple factorial model which incorporated measurements of P availability, feed efficiency and normal whole-body P concentration indicated that the dietary requirement was approximately 10 g kg⁻¹. The dietary requirement established in this study (10–11 g kg⁻¹) is higher than previously reported for Atlantic salmon or other fishes. Possible reasons for the wide range of reported dietary P requirements in fishes are discussed.     

Target spawning requirements for Atlantic salmon, Salmo salar L., in Newfoundland rivers

Abstract In recent years, there has been increasing pressure from user groups in Newfoundland and Labrador, Canada, to manage Atlantic salmon, Salmo salar L., on a river-specific basis. The requirement to maximize benefits to users while ensuring that conservation goals are met has resulted in an initiative to develop river-specific target spawning requirements. In Newfoundland, anadromous juvenile Atlantic salmon make widespread use of lacustrine habitat for rearing. Depending on the river, lacustrine habitat can account for a substantial proportion of total production. Therefore, target spawning requirements for a number of rivers in several management areas were derived in terms of the contribution of both riverine and lacustrine habitats. The approach adopted, and derivations of associated parameter values, are reviewed, and limitations and research needs are discussed.     

Infectious pancreatic necrosis in Atlantic salmon, Salmo salar L

A clinical and histopathological review was carried out of 21 outbreaks of acute infectious pancreatic necrosis (IPN) in Scottish Atlantic salmon, Salmo salar L., farms (13 marine and eight fresh water) during 1991-2004. A distinctive syndrome was evident in both post-smolts in sea water and fry in fresh water, where liver lesions, which had not previously been associated with IPN, became a consistent finding in addition to the more typical pancreatic and intestinal changes. Initial cases were described in post-smolts in Shetland, but by the end of the period of investigation this type of pathology had extended down the West coast of Scotland and into Ireland. Limited viral strain analysis suggested that similar strains were involved in both fresh water and sea water and that these differed from earlier isolates from rainbow trout, Oncorhynchus mykiss (Walbaum). In fresh water, recovered fish frequently developed a greatly distended intestine associated with accumulation of undigested food. In sea water, after the initial, often significant (50% or more), losses, there were many fish which failed to grow and became chronically emaciated and prone to sea louse infection. Although use of transfer diets containing immune enhancers and the selection of IPN resistant broodstock has reduced losses the disease remains a serious cause of economic loss.     

Assessment of infectious salmon anaemia virus prevalence for different groups of farmed Atlantic salmon, Salmo salar L., in New Brunswick

Abstract Infectious salmon anaemia (ISA) virus (ISAV) has been causing disease in New Brunswick since 1996. As a control measure, all fish in an outbreak cage are killed. The objective of this study was to compare ISAV prevalence in cages experiencing an outbreak with healthy cages from the same farm, neighbouring farms and distant farms. Atlantic salmon from five different groups were tested using an RT-PCR test. Groups included moribund fish from a cage experiencing an outbreak (A), healthy fish from an outbreak cage (B), healthy fish from a negative cage from a farm experiencing an outbreak in a different cage (C), healthy fish from a negative farm near an outbreak farm (D), and healthy fish sampled at a negative farm located in an area with only negative farms (E). Apparent prevalences (standard error) for the different groups (A-E) were 0.94 (+/-0.026), 0.41 (+/-0.062), 0.29 (+/-0.040), 0.08 (+/-0.037) and 0.08 (+/-0.037), respectively. All groups were significantly different (P < 0.002) from each other except for groups B and C and groups D and E. Because the prevalence of the virus was significantly higher in the outbreak cage (B) compared with other sites, early harvest of outbreak cages will remove one source of virus. However, ISA negative cages (C) that remain on the positive farm may potentially act as a viral reservoir.     

Content of synthetic astaxanthin in escaped farmed Atlantic salmon, Salmo salar L., ascending Norwegian rivers

Abstract  Isomeric ratios of astaxanthin in eggs and alevins of Atlantic salmon, Salmo salar L., have proven useful in identifying female spawners of farmed origin, but the method underestimates the proportion of fish of farmed origin. The rate of underestimation was studied by analysing astaxanthin content in tissue of 55 farmed Atlantic salmon ascending two Norwegian rivers in the autumn of 1991. The astaxanthin content fell into two distinct classes. Fifty-one per cent of the adult escaped salmon had isomeric ratios similar to salmon fed synthetic astaxanthin, whereas all the remaining fish had ratios similar to wild fish. Discriminant analysis classified 96.4% of the fish with known astaxanthin content into the correct astaxanthin class on basis of tail-fin erosion, length, weight and gill-cover damage. This discriminant function was used to estimate the astaxanthin classification of 1017 farmed salmon caught in nine rivers during 1989–1991. The classification success varied among years from 52 to 64%. Corresponding numbers for females and males were 45–48% and 54–70%, respectively. Thus, estimates of spawning rates of farmed female salmon via astaxanthin content in eggs or alevins from redds should be adjusted accordingly. The observed isomeric ratios of astaxanthin in the escaped farmed salmon and the relationship with morphology indicates that a significant proportion of the escapees ascending rivers have spent more than 1 year in the wild after escape.     

An estimation of dietary iron requirement of Atlantic salmon, Salmo salar L., parr

A study was conducted to determine the dietary iron requirement of fingerling Atlantic salmon Salmo salar L. During the first 4 weeks of the experiment, fish with an initial weight of 5 g were fed a casein–gelatine-based purified diet which contained 11 mg iron kg^?1. Thereafter duplicate tanks (200 fish in each) were fed the casein–gelatine purified diets containing supplemental iron levels of 0, 10, 20, 30, 40, 60, 100, 200 or 400 mg iron kg^?1 (added as FeSO_?4* 7H₂O) for 12 weeks. Weight gain, body length and mortality were monitored. Liver iron and ascorbic acid concentration were analysed in addition to whole-body iron, manganese and zinc concentration. Several haematological parameters were also measured. There were no significant differences in weight gain and survival of salmon fed diets containing different iron levels. Haematological values, hepatic and whole-body iron concentrations were, however, significantly affected by the dietary iron content. Liver vitamin C concentration decreased with increasing dietary iron levels. Dietary supplementation with iron significantly reduced whole-body manganese, but no effect of dietary iron on whole-body zinc was found. Based on haematology and hepatic iron concentration, the iron requirement of Atlantic salmon was determined to be between 60 and 100 mg iron kg¹.