The immunomodulatory effect of laminaran [β(1,3)-D-glucan] on Atlantic salmon, Salmo salar L., anterior kidney leucocytes after intraperitoneal, peroral and peranal administration

Anterior kidney leucocytes obtained from Atlantic salmon, Salmo salar L., 2 days after administration of laminaran, were assayed for their capacity to reduce nitroblue tetrazolium to formazan, and for their activity of lysosomal acid phosphatase after intraperitoneal (15 mgkg^?1), peroral (150 mg kg^?1) or peranal (150 mg kg^?1) administration. Leucocytes obtained from salmon treated by an intraperitoneal injection of laminaran produced significantly more superoxide anion than cells obtained from fish treated with dextran or sodium chloride immediately after cell isolation. Immediately after extraction, the activity of acid phosphatase in anterior kidney leucocytes obtained from salmon injected with laminaran was significantly higher than in cells harvested from fish treated with dextran or sodium chloride. Furthermore, cells obtained from salmon treated by peroral instillation of laminaran showed significantly enhanced production of superoxide anion compared with leucocytes from fish treated with either sodium chloride or dextran. The acid phosphatase activity in anterior kidney leucocytes from salmon treated by peroral and peranal instillation of laminaran was significantly higher than in cells from fish treated either with sodium chloride or dextran. Finally, fluorescence microscopic examination of tissue sections from fish treated peranally by intubation with fluorescein labelled laminaran revealed fluorescent vesicles in intestinal epithelial cells and in anterior kidney macrophages.     

Accumulation of immunomodulatory laminaran [β(1,3)-D-glucan] in the spleen and kidney of Atlantic salmon, Salmo solar L.

Abstract. The tissue distribution of ³H- and FITC-laminaran in Atlantic salmon, Salmo salar L., was investigated after intravenous administration. Liquid scintillation counting and whole-body autoradiography revealed that the concentration of ³H-labelled laminaran in the spleen and anterior kidney was higher than in blood throughout the experimental period (1 h to 12 days). Renal excretion and intestinal exsorption were the main elimination pathways for laminaran. Microscopic examination revealed accumulation of FITC-labelled laminaran in macrophages in kidney and spleen. In addition, endothelial cells in the kidney, spleen, liver and intestine contained the immunomodulator. Thus, these experiments show that laminaran accumulates and is retained in immunologically relevant cells in the kidney and spleen.     

Intestinal absorption of immunomodulatory laminaran and derivatives in Atlantic salmon, Salmo salar L.

Abstract. Radiolabelled (¹²⁵I, ³H) immunomodulatory laminaran (isolated from Laminaria hyperborea ), a β(1,6)-branched β(1,3)-D-glucan and different radiolabelled sulphated analogues of laminaran were administered peranally to Atlantic salmon, Salmo salar L. Intestinal absorption and tissue distribution were examined by means of radioactive tracer techniques. The intestinal uptake was highest with native laminaran and laminaran with a low degree of sulphatation, while highly sulphated laminarans were poorly absorbed. The tissue distribution analysis revealed high amounts of radiolabelled compound in the liver, and anterior and posterior kidney, whereas the spleen contained low amounts. Peak serum and organ concentrations were reached about 30 min after administration. The results were confirmed by autoradiography of tissue sections from salmon after peranal administration of radiolabelled laminaran. Finally, the peranal administration of fluorescence labelled laminaran revealed epithelial supranuclear fluorescent vacuoles containing laminaran. It is concluded that native laminaran and slightly sulphated laminaran are absorbed from the posterior intestine and that they are distributed to tissues rich in immunocompetent cells. Thus, these compounds may have potential as immunomodulatory feed additives.     

Accumulation of immunomodulatory laminaran (β(1,3)-D-glucan) in the heart, spleen and kidney of Atlantic cod, Gadus morhua L.

The tissue distribution of ³H- and FITC-lammaran in Atlantic cod, Gadus morhua L., was investigated after intravenous administration. Liquid scintillation counting and whole-body autoradiography revealed a substantial accumulation of ³H-labelled laminaran in the heart throughout the experimental period (1 h to 12 days). High amounts of the immunomodulator were also present in the spleen, anterior kidney and posterior kidney during the study. Renal excretion and intestinal exsorption were the main elimination pathways. Fluorescence microscopic examination revealed accumulation of FITC-labelled laminaran in macrophages in the kidney and ellipsoid sheath cells (macrophages and reticular cells) in the spleen. Furthermore, endothelial entrapment in the heart was pronounced. Endothelial cells in the kidney also contained the FITC-labelled immunomodulator. Immunohistochemical analysis, using anti-β(l,3)-D-glucopyranose antibody, also showed endothelial uptake of laminaran in the heart. The present investigation shows that laminaran accumulates and is retained in immunologically relevant cells in the Atlantic cod.     

Atlantic salmon, Salmo salar L., feed in Scottish coastal waters

Abstract. The stomach contents of 256 Atlantic salmon, Salmo salar L., of fork lengths 53–66cm, caught in bag nets on the Scottish west coast near Ullapool, were examined between June 1983 and July 1986. A total of 61 fish contained fish in the stomach or had faecal pellets containing fish bones in the gut. All recognizable whole fish were sandeels, Ammodytes marinus Raitt, ranging in size from 4.5 to 15cm. No evidence for crustacean or other non-fish prey items was found. Results indicate that feeding salmon were caught up to a certain cut-off point in June or early July, after which all salmon sampled were not feeding. It is suggested that either there is a local feeding stock of salmon or that fish feed during migration from the Faroe Isles or other possible distant water origins.     

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.     

Mitochondrial and peroxisomal β-oxidation capacities in various tissues from Atlantic salmon Salmo salar

In order to investigate the capacities of different tissues to oxidize fatty acids, total β-oxidation (mitochondrial and peroxisomal) of [1–¹⁴C]palmitoyl-CoA was determined in liver and red- and white muscle from adult and juvenile Atlantic salmon Salmo salar. By including potassium cyanide (KCN) in the assay medium, it was possible to differentiate between mitochondrial and peroxisomal β-oxidation capacities. Mitochondrial β-oxidation dominated in all tissues except in livers from juvenile fish where the peroxisomal β-oxidation dominated. In general, the red muscle possesses the highest fatty acid oxidation capacity, however, by taking into consideration the fact that white muscle occupies approximately 60% of the total body weight, this study demonstrates that the white muscle is an important tissue in the overall fatty acid catabolism.     

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.     

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.     

Prophylactic effect of β(1,3)-D-glucan (laminaran) against experimental Aeromonas salmonicida and Vibrio salmonicida infections

Immersion and intraperitoneal vaccination are the two most common and successful methods for vaccination of fish (Holm & Júrgensen 1987; Lillehaug 1989). Effective vaccines and the improvement of environmental conditions have dramatically reduced the amount of antibiotics used in aquaculture in Norway from 48.5 t in 1987 to 1.03 t in 1996. Nevertheless, the labour intensive process of vaccination and the often undesirable side-effects of the adjuvants incorporated in injectable vaccines (Anderson 1992; Mulvey, Landolt & Busch 1995; Midtlyng, Reitan, Speilberg 1996), have resulted in a search for alternative approaches to achieve immune stimulation. Hence. there is growing interest in the development of immunomodulators that do not cause or have only minor unwanted effects on the host and preferably can be given as a feed additive. It is widely acknowledged that the beneficial (or in some instances undesirable) effects of immunomodulators such as b-glucans and bacterial products in an infection are mainly a result of their stimulatory effects on macrophages (Johnston 1978; Chung & Secombes 1988; Júrgensen, Lunde & Robertsen 1993).     

Isolation of salmon pancreas disease virus (SPDV) from farmed Atlantic salmon, Salmo salar L., in Scotland

A toga-like virus was recently isolated in the present authors' laboratory from Atlantic salmon during an outbreak of PD in Ireland (Nelson, McLoughlin, Rowley, Platten & McCormick 1995). When inoculated into Atlantic salmon post-smolts, this virus, known as salmon pancreas disease virus (SPDV), caused pathological changes in the pancreas, heart and skeletal muscle which were indistinguishable from those observed in natural outbreaks of PD (McLoughlin, Nelson, Rowley, Cox & Grant 1996). In 1996 a serological survey was carried out to determine the presence and distribution of antibodies to SPDV in Irish, Scottish and Norwegian farm-reared Atlantic salmon (McLoughlin, Rowley & Doherty 1998). The results of this survey indicated that SPDV is present in the major European salmon-producing countries. The present paper describes the first isolation of SPDV from farm-reared, PD-affected Atlantic salmon in Scotland.     

Incidence of hybridization between Atlantic salmon, Salmo salar L., and brown trout, Salmo trutta L., in Ireland

The substantial growth of the farmed salmon industry in Europe since the 1970s has highlighted concerns regarding the genetic impact of escaped farmed salmon on wild salmonid stocks. High incidences of salmon × trout hybrids have been recorded in rivers situated near intensive salmon farming in Norway and Scotland, which may be indicative of a breakdown in reproductive isolation between salmon, Salmo salar L., and brown trout, Salmo trutta L. In the present study, salmonid fry and 0+ parr were collected from rivers in western Ireland. Allozyme and minisatellite DNA analysis were carried out on fry to determine the frequency of F1 hybrids from 10 rivers located within 38 km of salmon farms and three rivers at least 80 km from salmon farms. A total of 49 hybrids were recorded from 4135 salmonid fry (frequency = 1.2%). Mitochondrial DNA analysis showed that all hybrids arose from Atlantic salmon female × brown trout male crosses. Hybrid parr were recorded from one of the low-risk rivers (1.0%), but were present in seven out of the 10 catchments located within 38 km of salmon farms, with frequencies ranging from 0.7% to 3.1%. The results of the present survey, which represents the first extensive record of the levels of salmon-trout hybridization in Ireland, are discussed in relation to the continued growth of salmon farming in this country.     

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.