A historical review of fish vaccinology.

The first report on fish vaccination to appear in a widely read international scientific journal was that by Duff [5] dealing with his results obtained with anti-furunculosis vaccines. Duff's report did not result in an immediate landslide of fish vaccination trials in other laboratories because, in the years following the second world war, the preoccupation was with disease control using the newly discovered antibiotics. In fish culture, the ensuing 30 to 40 years might accurately have been termed the "era of chemotherapy" because large numbers of antibiotics, sulpha drugs, and even mercury-based antimicrobial agents were routinely used. It was only in the mid to late 1970s, with an increased interest in fish farming, particularly marine fish farming, that attention was once again turned to the possibility of vaccination as a means of preventing/ controlling fish diseases and to the development of commercially available vaccines. The reasons for this turn of events were varied: the high cost of using chemotherapy, the short-term nature of the protection obtained with antibiotics, the increasing appearance of antibiotic resistant fish pathogens, and, to some extent, concerns about the environmental impacts of antibiotic use. This paper briefly outlines the success that has attended efforts to develop vaccines against some of the more important bacterial diseases of cultured fish and the progress made in developing vaccines against important viral fish pathogens. In the process, an attempt will be made to show how fish vaccine development has benefited from an improved knowledge of the fish's immune system and from a better understanding of the virulence factors possessed by particular fish pathogens.

A field evaluation of an indirect fluorescent antibody-based broodstock screening test used to control the vertical transmission of Renibacterium salmoninarium in Chinook salmon (Oncorhynchus tshawytscha).

Ovarian fluid samples from erythromycin treated and untreated spawning three year old Chinook salmon were screened independently by two laboratories for the presence of Renibacterium salmoninarum using the indirect fluorescent antibody technique (IFAT). Agreement between the results of the two laboratories could be explained by chance when R. salmoninarum cell numbers as low as one per sample were considered sufficient to represent a positive result. If a positive result was considered to be the detection of larger numbers of R. salmoninarum cells (greater than 51 cells per sample), agreement increased and there was a statistically significant association between the results of the two laboratories. However, the level of agreement did not reach satisfactory levels for a population screening test. Furthermore, approximately 60% of the samples yielded false negative results when IFAT results were compared with positive culture results. These results led to the conclusion that the IFAT screening procedure, as carried out, was unsuitable for the purposes intended. Erythromycin injection of the spawning fish had no statistically significant effect on the results of the IFAT screening test.

Acquired and innate resistance to the haemoflagellate cryptobia salmositica in sockeye salmon (Oncorhynchus nerka).

Juvenile sockeye salmon (Oncorhynchus nerka, Fulton River stock) were protected from otherwise lethal challenges with the haemoflagellate Cryptobia salmositica by acclimation to elevated water temperatures (20 degrees C). Fish treated in this manner displayed increased immunity to C. salmositica and yielded plasma showing enhanced lytic activity against the parasite. The acquired lytic activity was antibody- and complement-mediated. In contrast, a stock of naive O. nerka from Weaver Creek, previously identified as having a high innate resistance to the lethal effects of C. salmositica, also had plasma factors that destroyed the parasite in vitro. This anti-Cryptobia activity also involved complement because 1) it resulted in lysis of the parasite, 2) it was heat-labile (40 degrees C for 20 min), and 3) it was largely removed from the plasma by substances capable of activating (binding) complement by the classical pathway (an antigen:antibody complex of Renibacterium salmoninarum and its specific antibody) and the alternate pathway (Escherichia coli lipopolyssacharide). The complement-mediated lysis associated with innate resistance was apparently the result of activation by the alternate pathway because it occurred in fish lacking antibodies against the parasite. The reaction was unusual in that a long incubation period (about 2 days) was required for maximum lysis of the parasite. At least one component of the innate lytic system depended on disulphide bonds because lytic activity was destroyed by 2-mercaptoethanol.

Immunity to Aeromonas salmonicida in coho salmon (Oncorhynchus kisutch) induced by modified Freund's complete adjuvant: its non-specific nature and the probable role of macrophages in the phenomenon.

Juvenile coho salmon (Oncorhynchus kisutch), vaccinated with one intraperitoneal injection of formalin-killed virulent Aeromonas salmonicida cells suspended in saline, showed increased protection against approximately one LD60 of homologous challenge administered at 30 days post-vaccination. Under similar conditions, coho vaccinated with a modified complete Freund's adjuvant (MFCA) alone were also equally protected. When measured against a more severe A. salmonicida challenge of approximately one LD95, the strength of the MFCA-induced protection was found to exceed that produced by the homologous bacterin administered in saline or incomplete adjuvant, and the protection was still evident at 90 days post-treatment. Other more precise measurements indicated the LD50 for MFCA-treated coho to be up to 450 times that for saline-treated coho. Two other tested adjuvants, levamisole and MDP (N-acetyl-muramyl-L-alanyl-D-isoglutamine), administered in a modified Freund's incomplete adjuvant, also enhanced anti-A. salmonicida immunity but to a lesser degree. The active factor in MFCA was a killed Mycobacterium butyricum preparation, and the anti-A. salmonicida immunity it induced was non-specific because the immunity extended to two other serologically distinct fish pathogens tested: A. hydrophila (LD50 increase of 5.3-fold) and Vibrio ordalii (LD50 increase of 560-fold). Macrophages are believed to account for the M. butyricum-induced anti-A. salmonicida immunity because the immunity was a) non-specific, b) very rapid in onset (it was measurable by 4 days), and c) influenced by particulate preparations, known to affect macrophage function and immunity in mammals. The possible benefits of adjuvant-induced non-specific immunity in cultured fish are discussed.