ALBERT

Notes: Abstract. Protection against Aeromonas salmonicida was determined by passive immunization and with various bacterin preparations. Rabbit antiserum was prepared against a rough, virulent strain of A. salmonicida (AS-1R), the same strain boiled (AS-1R, boiled), and an avirulent, smooth strain of this same isolate (AS-1S). Cross-absorption, cross-passive protection and analysis by counter immunoelectrophoresis of various extraction methods were studied. It was shown that AS-1R cells contained an additional antigen not present in AS-1R (boiled) and AS-1S cells. Antiserum to the AS-1R antigen passively protected sockeye salmon, Oncorhynchus nerka (Walbaum), against a virulent challenge, and antisera to AS-1R (boiled) and AS-1S were not protective. The antigen was not destroyed by formalin or heat at 5°C for 60 min, but it appeared to be partially inactivated with proteolytic enzymes. The antigen was produced in casein yeast beef (CYB) broth up to 32 h but not thereafter, and low yields were obtained in tryptic soy or brain heart infusion (BHI) broth. It was extracted from cells with ethylenediamine tetraacetic acid (EDTA) and especially alkaline hydrolysis, but not with proteolytic enzymes or detergents. The detergents appeared to destroy the antigen. We concluded that the antigen was protein and is most likely the external A-protein (AP) reported for rough, virulent strains of A, salmonicida. Various methods of preparing A. salmonicida bacterins were evaluated by determining the level of protective immunity induced in intraperitoneally (i.p.) vaccinated fish. Growth of cells in CYB or BHI broth resulted in production of only rough (autoagglutinated in saline) variants of A. salmonicida. Although only rough variants were associated with protective immunity, one strain was not protective, it was avirulent by bath challenge. Bacterins prepared in CYB were more efficacious than those grown in BHI, but inactivation with formalin, iodine, or glutaraldehyde worked equally well. However, boiling the bacterin or filtering the cells from the bacterin removed its efficacy. Methods of releasing the AP were evaluated by sonification, pH-lysis, disaggregation and treatment with EDTA, and all treatments worked equally well. Also, precipitation on to aluminium or use of Freund's complete adjuvant did not significantly improve the protection. In parenterally vaccinated fish, protection was demonstrated by challenging the fish at various levels by bath, injection or cohabitation with infected fish. The best protection was demonstrated using the cohabitation challenge method. The potency and field efficacy of an A. salmonicida bacterin prepared in CYB broth and extracted with 5 mM EDTA was evaluated. Fish were vaccinated by i.p, injection and potency was determined in the laboratory by experimental challenge and in the field by natural challenge. Chinook salmon, O.ishawytschu (Walbaum), developed immunity within seven days at 10°C. The bacterin could be diluted up to 1:2000 without loss of potency. The field tests results were equivocal; however, (he prevalence of infection was lower in vaccinated fish.

Notes: Abstract. The comparative efficacy of several methods of vaccinating rainbow trout, Salmo gairdneri Richardson, with Yersinia ruckeri bacterins was evaluated. In general, the protective immunity was best by intraperitoneal injection followed by direct immersion, shower and spray, respectively. The bacterin was effective by all methods when diluted 1:20 or less, but there was a trend towards less efficacy with higher dilutions and shorter exposure time.

Notes: Abstract. The level of protective immunity was determined for several salmonid species following vaccination by the direct immersion method with commercial Vibrio anguillarum (two serotypes) and Yersinia ruckeri (Hagerman strain) bacterins. The duration of protective immunity varied with the bacterin concentration, size and species of fish, but the duration between the two bacterins was comparable. In fish under 1 g duration of protective immunity was longest when the most concentrated bacterin was used. Generally, immunity lasted in 1-g fish for about 120 days, in 2-g fish for about 180 days, but in 4-g fish and larger immunity lasted for a year or longer. Coho salmon (Oncorhynchus kisutch) and sockeye salmon (O. nerka) retained immunity for a longer time and pink salmon (O. gorbuscha) the shortest time. Chinook salmon (O. tshawytscha) and rainbow trout (Salmo gairdneri) were intermediate. Field data generally followed the laboratory tests, but the duration appeared somewhat shortened. In one test 20-g rainbow trout were vaccinated by the shower method and no loss of immunity occurred after 311 days.

Notes: Abstract. The fry of several salmonid species were vaccinated by direct immersion in either Yersinia ruckeri or Vibrio anguillarum bacterin and the level of protective immunity was determined by the survival of fish after bath challenge with virulent organisms. The immersion time for effective vaccination was obtained within 5 s and protective immunity was demonstrated within 5 days at 18°C and within 10 days at 10°C. The minimum size at which maximum protective immunity occurred was between 1.0 and 2.5 g. Immunity appeared to be a function of size and not age, but differences in response among several species were indicated. In fish under 1.0 g, the level of protective immunity could be increased using a more concentrated bacterin. The results were similar with both bacterins.

Notes: Abstract. Several factors affecting the potency of Yersinia ruckeri bacterins were evaluated by vaccinating rainbow trout, Salmo gairdneri Richardson, with various bacterins using the immersion method and determining the level of protective immunity after a virulent challenge. The potency of bacterins prepared with tryptic soya broth at room temperature was not affected by growth at pH from 6.5 to 7.7 or by a culture age from 9 to 96 h. Chloroform and formalin inactivated (0.3%) bacterins gave comparable results and no enhancement of potency occurred by prior extraction of bacterial cells with either butanol or phenol. Cell lysis, as measured by reduced optical density, occurred when cells were held at pH 9–8 for 60 to 120 min. Bacterins prepared from pH-lysed cells resulted in a significant increase in protective immunity. Bacterins prepared at pH 7.2 for 48 h, pH-lysed and inactivated with 0.3 % formalin could be diluted up to 1:100 without loss of efficacy when applied to rainbow trout by a single 20 s immersion. However, with bacterin diluted 1:10 loss of potency occurred after 20 consecutive immersions (100 kg of fish) in the same bacterin at a rate of 0.5 kg/1 of diluted bacterin for each immersion. Factors affecting optimum duration of immunity are discussed.

Notes: Abstract. A serological study of 23 North American strains of Yersinia ruckeri showed that they were comprised essentially of one serotype–the classical ‘Hager-man’ serotype. Cross-protection tests showed that rainbow trout could be protected from challenge with various strains. Continuous surveillance of enteric redmouth disease and further work including DNA binding studies is necessary to determine the precise significance of the other related serotypes.

Notes: Abstract. Bacterins of Vibrio anguillarum, Yersinia ruckeri, Aeromonas Salmonicida, and Renibacterum salmoninarum were administered alone and in combination to salmonid fishes and the level of protective immunity compared. With each pathogen, the protection obtained with monovalent with A. salmonicida bacterin alone conferred some protection against challenges with Type II V. anguillarum and Y. ruckeri. Also, the combination of A. salmonicida and R. salmoninarum bacterins appeared to potentiate the protection conferred against A. salmonicida. The potential of multivalent vaccines for protecting fish from several diseases appears to be real.

Notes: Abstract. The efficacy of the immersion method of vaccination was evaluated using Aeromonas salmonicida bacterin. In general a 2 min immersion vaccination was not as effective as vaccination by intraperitoneal injection; however, the level of immunity was improved by giving multiple vaccinations several days apart. A primary immersion vaccination with bacterin diluted 1:4 followed by a secondary vaccination diluted 1:2 gave good results. The most concentrated secondary booster was more effective than boosting with more dilute bacterin.