Lack of evidence for vertical transmission of SAV 3 using gametes of Atlantic salmon, Salmo salar L., exposed by natural and experimental routes.

Pancreas disease (PD) is an important cause of losses in farmed salmonids in Norway, the United Kingdom and Ireland. As the spread of salmonid alphavirus (SAV), the causal agent, to naïve populations is of major concern to the farming industry, it is important to uncover the transmission routes of the virus. This study was conducted to investigate the potential for vertical transmission of SAV subtype 3. Progeny of broodstock with signs of late-stage PD and persistent RT-PCR signals for SAV were followed from fertilization to smoltification in an experimental facility. Fertilized ova were either not disinfected or taken through one of three different disinfection regimes. Also, ova and milt from uninfected broodfish from a different population were exposed to a cell-cultured strain of SAV 3 immediately before fertilization to simulate a viraemic phase in parent fish. A group of uninfected controls were also included in the study. Fertilized ova from bath exposed and negative control groups were double disinfected. Following fertilization, experimental fish went through a normal freshwater phase. However, fry were stressed at first feeding to enhance replication of possibly latent virus. Smoltification was induced by an artificial light regime, and experimental fish were followed to the late smoltification phase. Selected samples were investigated by real-time RT-PCR for SAV, by histology for evidence of PD and by serology for neutralising antibodies against SAV. All analysed samples of progeny were negative. This result shows that SAV 3 is not readily transmitted vertically from parents to offspring. Additional negative PCR results from salmon sampled in commercial hatcheries support these findings. Also, recent studies have shown that risk factors for the horizontal transmission route explain the vast majority of PD outbreaks in Norway. It is concluded that if it happens at all, vertical transmission is of minor importance in the spread of SAV 3.

Risk factors for pancreas disease (PD) outbreaks in farmed Atlantic salmon and rainbow trout in Norway during 2003-2007.

Pancreas disease (PD) is an emerging infectious disease in farmed Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss Walbaum) caused by salmonid alphavirus (SAV). The present study is a large scale study aiming at quantifying the probability of contracting PD in farmed salmonid cohorts in Norway due to exposure to risk factors that may be associated with specific transmission pathways for SAV, or may increase a cohort's susceptibility to PD. Monthly reports of numbers of fish and mean fish weight from all marine salmonid farm sites in Norway were used to identify cohorts of farmed salmonids. Only cohorts that were initiated and terminated during 2003-2007 were assembled for the study. Records of clinical diagnosis of PD on marine farm sites were used to identify PD case cohorts. In PD case cohorts, PD-outbreaks were defined to start the month the diagnosis was recorded and last until the cohort was terminated. All cohorts in which PD was not recorded were assigned to the control-class. In total 143 PD case cohorts and 1079 control cohorts were assembled. Risk factors were assigned to the cohorts and analysed using logistic regression by generalized additive models (GAM). We find that infection pressure, a variable designed to capture the potential for local disease spread, has a strong effect on the probability of recording a PD-outbreak in a cohort. The function describing the effect of infection pressure increased steeply as infection pressure increased from 0 to moderate values corresponding to having a mean sized neighbouring fish stock with PD at a distance of 2 km, after which the function levelled off. The study emphasises horizontal transmission pathways as important for the spread of PD in Norwegian salmon farming, and accordingly that bio-security measures aimed at controlling horizontal transmission are necessary in order to reduce the number of outbreaks of PD.

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.

Experimental transmission of cardiomyopathy syndrome (CMS) in Atlantic salmon Salmo salar.

Cardiomyopathy syndrome (CMS) is a disease of unknown aetiology, having significant economic impact as it primarily affects large, farmed Atlantic salmon Salmo salar L. in seawater, close to harvest. In the present study, we have demonstrated that CMS is a transmissible disease under experimental conditions. Histopathological lesions consistent with CMS were induced in Atlantic salmon post-smolts after injection of tissue homogenate from farmed fish diagnosed with CMS. Six weeks post-injection (p.i.), experimental fish started developing focal to multi-focal lesions in the atrial endo- and myocardium, with subsequent progression to the ventricle. This proceeded into severe endocarditis and subsequent myocarditis with mononuclear cell infiltration of the atrium and, to a lesser degree, the spongy layer of the ventricle. These lesions were consistent with histopathological findings in field outbreaks of CMS. From Week 33 p.i., lesions also appeared in the compact myocardium, with focal epicarditis adjacent to focal myocardial lesions. In conclusion, these results indicate that CMS has an infectious aetiology and should be treated as a potentially contagious disease.

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.

Pathology of heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon Salmo salar.

This is the first description of heart and skeletal muscle inflammation (HSMI), a novel disease affecting farmed Atlantic salmon Salmo salar in Norway. HSMI was first diagnosed in 1999, and there has since been a yearly increase in the number of recorded outbreaks. Atlantic salmon are commonly affected 5 to 9 mo after transfer to sea, but outbreaks have been recorded as early as 14 d following seawater transfer. Affected fish are anorexic and display abnormal swimming behaviour. Autopsy findings typically include a pale heart, yellow liver, ascites, swollen spleen and petechiae in the perivisceral fat. While mortality is variable (up to 20%), morbidity may be very high in affected cages. Until more accurate tests are available, HSMI is diagnosed on the basis of histopathology. The major pathological changes occur in the myocardium and red skeletal muscle, where extensive inflammation and multifocal necrosis of myocytes are evident. HSMI is transmissible and, although most likely caused by a virus, the causal agent has not yet been isolated. This paper describes clinical signs and pathology of HSMI from 3 field outbreaks in Norway. Microscopic lesions are compared and discussed in relation to published descriptions of pancreas disease (PD) and cardiomyopathy syndrome (CMS). It is concluded that HSMI is histopathologically distinguishable from PD and CMS.

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.