Trygve T. Poppe

Vaccination-Induced Systemic Autoimmunity in Farmed Atlantic Salmon

Over half of the salmon consumed globally are farm-raised. The introduction of oil-adjuvanted vaccines into salmon aquaculture made large-scale production feasible by preventing infections. The vaccines that are given i.p. contain oil adjuvant such as mineral oil. However, in rodents, a single i.p. injection of adjuvant hydrocarbon oil induces lupus-like systemic autoimmune syndrome, characterized by autoantibodies, immune complex glomerulonephritis, and arthritis. In the present study, whether the farmed salmon that received oil-adjuvanted vaccine have autoimmune syndrome similar to adjuvant oil-injected rodents was examined. Sera and tissues were collected from vaccinated or unvaccinated Atlantic salmon (experimental, seven farms) and wild salmon. Autoantibodies (immunofluorescence, ELISA, and immunoprecipitation) and IgM levels (ELISA) in sera were measured. Kidneys and livers were examined for pathology. Autoantibodies were common in vaccinated fish vs unvaccinated controls and they reacted with salmon cells/Ags in addition to their reactivity with mammalian Ags. Diffuse nuclear/cytoplasmic staining was common in immunofluorescence but some had more specific patterns. Serum total IgM levels were also increased in vaccinated fish; however, the fold increase of autoantibodies was much more than that of total IgM. Sera from vaccinated fish immunoprecipitated ferritin and ∼50% also reacted with other unique proteins. Thrombosis and granulomatous inflammation in liver, and immune-complex glomerulonephritis were common in vaccinated fish. Autoimmunity similar to the mouse model of adjuvant oil-induced lupus is common in vaccinated farmed Atlantic salmon. This may have a significant impact on production loss, disease of previously unknown etiology, and future strategies of vaccines and salmon farming.

A colour atlas of salmonid diseases

Normal teleost histology autopsy procedures disease diagnosis viral diseases bacterial diseases fungal diseases parasitic diseases metazoa diseases due to nutritional imbalance tumour malformations environmental factors affecting fish health problems associated with poor husbandry disease of uncertain aetiology staining techniques for light microscopy formulae for fixatives, declarification and other solutions for light microscopy.

A novel totivirus and piscine reovirus (PRV) in Atlantic salmon (Salmo salar) with cardiomyopathy syndrome (CMS).

BackgroundCardiomyopathy syndrome (CMS) is a severe disease affecting large farmed Atlantic salmon. Mortality often appears without prior clinical signs, typically shortly prior to slaughter. We recently reported the finding and the complete genomic sequence of a novel piscine reovirus (PRV), which is associated with another cardiac disease in Atlantic salmon; heart and skeletal muscle inflammation (HSMI). In the present work we have studied whether PRV or other infectious agents may be involved in the etiology of CMS.ResultsUsing high throughput sequencing on heart samples from natural outbreaks of CMS and from fish experimentally challenged with material from fish diagnosed with CMS a high number of sequence reads identical to the PRV genome were identified. In addition, a sequence contig from a novel totivirus could also be constructed. Using RT-qPCR, levels of PRV in tissue samples were quantified and the totivirus was detected in all samples tested from CMS fish but not in controls. In situ hybridization supported this pattern indicating a possible association between CMS and the novel piscine totivirus.ConclusionsAlthough causality for CMS in Atlantic salmon could not be proven for either of the two viruses, our results are compatible with a hypothesis where, in the experimental challenge studied, PRV behaves as an opportunist whereas the totivirus might be more directly linked with the development of CMS.

Guidelines for health and welfare monitoring of fish used in research

The aim of this paper is to provide background material necessary for the development of international guidelines for the health and welfare monitoring of fish used in research. It provides an overview of present guidelines and discusses why more detailed and species-specific guidelines are needed. A major issue within fish research is to document the situation today and point out areas where improvements are needed.

Granulomatous Uveitis Associated with Vaccination in the Atlantic Salmon

This study addressed histologic and immunopathologic changes in ocular tissues and investigated the distribution of major histocompatibility class II (MHC class II)-positive cells in Atlantic salmon (Salmo salar) suffering from severe postvaccination disease. Twenty-nine fish with generalized inflammation, probably a result of vaccination, were investigated. One individual that had escaped vaccination was included in the study. Material was investigated by cultivation methods for fungi and bacteria. Histology using conventional staining procedures and immunohistochemistry with antisera against MHC class II β chain were performed. No growth was observed from the cultivation investigations. Histology revealed occlusion of the lumen in the larger choroid vessels and in the choriocapillaris, inflammatory infiltrations and loss of structure in the choroid rete, and, in some cases, aggregations of multinucleated giant cells (MGC) and Splendore-Hoeppli material. Immunohistochemistry demonstrated massive MHC class II+ cellular infiltrations in the uveal tract. Such infiltrations were also seen in the ventral ciliary cleft, a condition that is associated with glaucoma. Immunoreactive cells included dendritelike cells, epithelioid cells, and MGCs. The endothelia of smaller vessels were frequently MHC class II+, and immunoreactive infiltrations were seen in the optic nerve in several individuals. No pathologic changes were detected in the unvaccinated individual. In conclusion, generalized inflammatory reactions in fish may lead to severe ocular inflammation, occlusion of uveal vessels, and perivascular changes with MHC class II+ upregulation in cells in the uveal tract and optic nerve.

Ultrastructure of Spironucleus barkhanus N. Sp. (Diplomonadida: Hexamitidae) from Grayling Thymallus thymallus (L.) (Salmonidae) and Atlantic Salmon Salmo salar L. (Salmonidae)

ABSTRACT. A hexamitid flagellate Spironucleus barkhanus n. sp., from the lumen of the gut and gall bladder of wild grayling Thymallus thymallus, and from muscle abscesses of farmed Atlantic salmon Salmo salar from Norway, is described by light, scanning and transmission electron microscopy. The flagellate was axenically cultured in trypticase, yeast extract, iron serum medium. Live trophozoites from axenic cultures incubated at 5° C, measure 11–20 x 6–14 μm. The flagellates show a typical bi‐radial symmetry. Each recurrent flagellum is almost completely surrounded by a striated lamina. In the posterior end the lamina widens, appearing heart shaped in transverse section. Accompanying each recurrent flagellum are three narrow bands of microtubules, following the longitudinal groove created by the incomplete closure of the striated lamina. The recurrent flagella emerge posterio‐medially through cytostome openings halfway surrounded by crescent‐shaped ridges, oriented in opposite directions in the two openings. The position and adornment of the cytostome openings, and the arrangement and number of the microtubules accompanying the recurrent flagella, distinguish Spironucleus barkhanus n. sp. from previously described species of Spironucleus.

The effect of ploidy and incubation temperature on survival and the prevalence of aplasia of the septum transversum in Atlantic salmon, Salmo salar L.

Heart deformities are a concern in aquaculture and are linked to egg incubation temperature. Diploid and triploid Atlantic salmon, Salmo salar L., were incubated at 6, 8 and 10 °C and analysed for aplasia of the septum transversum (n = 150 ploidy⁻¹ incubation temperature⁻¹). Heart morphology (size and shape) was assessed in fish incubated at 6 °C and in fish with and without aplasia of the septum transversum (n = 9 group⁻¹) incubated at 10 °C. Egg mortality was significantly higher in triploids than in diploids at all incubation temperatures, and increased egg incubation temperatures increased mortality in both ploidy. Triploids grew quicker than diploids after egg incubation at 10 °C, but not at 6 °C. Aplasia of the septum transversum occurred only in triploid fish after incubation at 6 °C and 8 °C (0.7% and 3.3%, respectively) and was significantly greater (P ≤ 0.05) in triploids after incubation at 10 °C compared with diploids (30% and 18%, respectively). Aplasia of the septum transversum significantly increased heart mass and resulted in a long flat ventricle compared with fish displaying a septum transversum. The results suggest triploid salmon should be incubated below 8 °C.

Presence of Anisakis simplex (Rudolphi, 1809 det. Krabbe, 1878) and Hysterothylacium aduncum (Rudolphi, 1802) (Nematoda; Anisakidae) in runts of farmed Atlantic salmon, Salmo salar L

One hundred farmed Atlantic salmon, Salmo salar L., were examined for the presence of nematodes by digestion of tissue in HCl-pepsin solution. All fish were sampled from one cage in a fish farm on the Norwegian south-west coast. Fifty harvest quality salmon, that is, salmon for human consumption (mean 5.4 kg, variation 3.0-7.6 kg), were sampled at the processing line while 50 salmon runts (mean 1.1 kg, variation 0.4-1.8 kg), discarded due to poor performance, were sampled from the discard bin after the grading station. Runts are individual fish with clear signs of poor performance over time and abnormal appearance and are thus not processed for human consumption. No nematodes were found in the musculature or viscera of the 50 harvest quality salmon. In total, 75 nematodes were found in 10 (20%) of the runts; 53 nematodes in the viscera and 22 in the musculature. Nematodes in the musculature were identified as Anisakis simplex (Rudolphi, 1809 det. Krabbe, 1878), while nematodes in the viscera were identified as A. simplex and Hysterothylacium aduncum (Rudolphi, 1802).

Fluctuating asymmetry in farmed Atlantic salmon (Salmo salar) juveniles: also a maternal matter?

Developmental stability reflects the degree to which phenotypic expression is unaffected by random accidents or developmental noise. Developmental stability may be measured by phenodeviance or fluctuating asymmetry (FA), and estimation of developmental stability has attracted substantial interest because it appears to represent a relatively simple method to identify sub lethal stress exposure and to assess animal welfare. As a part of a long-term study, the work presented here primarily aimed to investigate impacts on developmental instability in farmed salmon offspring ten months post hatch attributable to maternal cortisol administration prior to spawning and mild hyperthermia exerted during incubation. Main results show that maternal cortisol enhancement increased the level of FA in pectoral and pelvic fins, but did not affect the frequency of malformations in offspring. Mild hyperthermia during incubation increased weight and fork length and also increased pelvic fin FA. Malformed fish were heavier and longer than the normal ones, and pelvic fin asymmetry was positively related to condition factor. These results illustrate plausible lasting impacts on offspring development due to the maternal endocrinological state at spawning and indicate that developmental instability in farmed salmon juveniles may mirror aspects of the broodstock’s housing conditions.

Polyclonal hypergammaglobulinemia and autoantibody production induced by vaccination in farmed Atlantic salmon.

The introduction of oil-adjuvanted vaccines in salmon aquaculture made large-scale production feasible by reducing the impact of infections. Vaccines given intraperitoneally (ip) contain oil adjuvant such as mineral oil. However, in rodents, a single ip injection of adjuvant hydrocarbon oil induces lupus-like systemic autoimmune syndrome. We have recently reported that autoimmune disease in farmed salmon, characterized by production of various autoantibodies, immune complex glomerulonephritis, liver thrombosis, and spinal deformity, are previously unrecognized side effects of vaccination. In the present study, we examined whether vaccination-induced autoantibody production in farmed Atlantic salmon is a mere result of polyclonal B-cell activation. Sera were collected from 205 vaccinated and unvaccinated Atlantic salmon (experimental, 7 farms) and wild salmon. Total IgM levels and autoantibodies to salmon blood cell (SBC) extract in sera were measured by ELISA and the relationship between hypergammaglobulinemia and autoantibody production was analyzed. Comparison of endpoint titers vs levels/units using a single dilution of sera in detection of autoantibodies to SBC showed near perfect correlation, justifying the use of the latter for screening. Both total IgM and anti-SBC antibodies are increased in vaccinated salmon compared with unvaccinated controls, however, they do not always correlate well when compared between groups or between individuals, suggesting the involvement of antigen-specific mechanisms in the production of anti-SBC autoantibodies. The primary considerations of successful vaccine for aquaculture are cost-effectiveness and safety. Vaccination-induced autoimmunity in farmed Atlantic salmon may have consequences on future vaccine development and salmon farming strategy. Evaluation for polyclonal hypergamamglobulinemia and autoimmunity should be included as an important trait when vaccine efficacy and safety are evaluated in future.