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Efficacy of mineral oil-based experimental injection vaccines against Flavobacterium psychrophilum were tested in rainbow trout, Oncorhynchus mykiss (Walbaum), under laboratory and field conditions. The vaccines consisted of formalin- or heat-inactivated whole bacterium cell preparations of two different serotypes (Fd and Th) or a combination of serologically different F. psychrophilum (Fd and/or Th and/or Fp(T);Th). Specific antibody responses against the bacterium in plasma and skin mucus were evaluated post-vaccination with enzyme-linked immunosorbent assay. Efficacy of the vaccinations was determined by challenge trials to F. psychrophilum with the vaccinated rainbow trout. Significantly higher antibody levels in plasma were detected in vaccinated fish compared with mock-vaccinated fish. Injection vaccination did not trigger specific antibody production in the skin mucus. Significantly higher survival of i.p. vaccinated fish compared with non-vaccinated fish was observed during the challenge. The results suggest that mineral oil-based injectable vaccines containing formalin- or heat-inactivated virulent cells of F. psychrophilum effectively triggered specific antibody production and protected the fish against bacterial cold water disease.

Aims: To investigate the relationships between water temperature, bacterial growth, virulence and antigen expression in Vibrio salmonicida, the causal agent of cold water vibriosis in Atlantic salmon (Salmo salar L.). Methods and Results:The significance of sea temperature was investigated using historical clinical and oceanographic data. An upper threshold for disease of approx. 10°C was established. The effects of culture temperature and media type on bacterial growth were studied on solid and in liquid media. The highest rates of cell division were identified at 15°C on solid media and 10°C in liquid media. Outer membrane protein (OMP) expression and serological response in Atlantic salmon were studied using sodium dodecyl sulphatepolyacrylamide gel electrophoresis, Western blotting and enzyme-linked immunosorbent assay. A novel 76-kDa OMP produced in unshaken cultures at 10°C was not found to stimulate a specific humoral response. Conclusions: Diagnostic agar plate-based incubation of suspected V. salmonicida should be carried out at 15°C. High yield broth cultures for vaccine production should be incubated at 10°C or lower. Significance and Impact of the Study: This study is, to the best of our knowledge, the first to identify different optimal temperatures in a bacterial species cultured on physically different types of media. The evidence presented suggests that V. salmonicida and possibly other bacteria destined for vaccine use in poikilothermic organisms should be cultured at temperatures consistent with that at which disease occurs.

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).