511
Disease Control
Vaccine Development Much effort has been expended on vaccine development,
with recent research highlighting major antigenic proteins of 7 kDa and 45 kDa
(Hirono et al. 1997 ). Programmes have included the use of passive immunisation
(Fukuda and Kusuda 1981a ), which is of dubious practical value, the more conventional approach of using formalin-inactivated whole-cell preparations (Kusuda and
Fukuda 1980 ; Fukuda and Kusuda 1981b ; Afonso et al. 2005 ) and the more modern
approaches of genetic engineering. Of relevance, the salinity of the growth medium
composition appears to have an effect on the subsequent immune response after
vaccination, with 2.5 % (w/v) rather than 0.5 % NaCl being the more effective
(Nitzan et al. 2004 ). Bacteriological media containing peptones, yeast extract and
salt led to the synthesis of a wider range of cellular components, (including novel
compounds of ~14 and ~21.3 kDa) than those produced in more in vivo type conditions. These compounds were recognised by post-disease sea bass serum
(Bakopoulos et al. 2003b ). It was demonstrated that administration of a formalininactivated preparation in Freund’s complete adjuvant by i.p. injection induced
agglutinating antibodies in yellowtail. Thus titres of 1:256–1:2048 were achieved 5
weeks after vaccination (Kusuda and Fukuda 1980 ). Vaccination enhances the nitric
oxide response, i.e. the production of reactive nitrogen intermediates with their antimicrobial activities, to infection with the pathogen, and is correlated with the level
of protection (Acosta et al. 2005 ). Further work, using a variety of vaccines and
application methods, demonstrated conclusively that fi sh could be protected against
subsequent infection by Ph. damselae subsp . piscicida, although this has been
refuted by some workers (e.g. Hamaguchi and Kusuda 1989 ). Toxoid enriched
whole cells applied by immersion led to a low antibody response an RPS of 37–41 %
in sea bream (Magariños et al. 1994c ). An improved RPS of >60 % after 35 days
resulted from use of an LPS mixed chloroform-killed whole cell vaccine (Kawakami
et al. 1997 ). Using formalin-inactivated cells with or without FCA and a range of
application methods, namely i.p. injection, 5–7 s spray, hyperosmotic infi ltration
and oral uptake via food, Fukuda and Kusuda ( 1981b ) reported encouraging results
within 21 days following artifi cial challenge with Ph. damselae subsp . piscicida.
The best results, conferring 100 % protection to the fi sh, were obtained by use of i.p.
injection or by spraying. The titre of agglutinating antibodies was measured at
between 1:4 and 1:128. A subsequent study by these authors has pointed to the value
of vaccinating with sub-cellular components, notably bacterial LPS (Fukuda and
Kusuda 1982 ). In this connection, a whole cell vaccine in combination with ECPs
was used more successfully than a commercial product by immersion for 1 h and
i.p. injection in sea bass (Bakopoulos et al. 2003a ). However, formalin-inactivated
whole cells administered intraperitoneally achieved an RPS of 96 % in sea bream
(Hanif et al. 2005 ). The question about the nature of the immune response after i.p.
vaccination with or without a booster after 4 weeks with a FIA adjuvanted inactivated whole cell vaccine was addressed by Arijo et al. ( 2004 ), who demonstrated a
Photobacterium damselae subsp. piscicida (= Pasteurella piscicida)
Disease Control
Vaccine Development Much effort has been expended on vaccine development,
with recent research highlighting major antigenic proteins of 7 kDa and 45 kDa
(Hirono et al. 1997 ). Programmes have included the use of passive immunisation
(Fukuda and Kusuda 1981a ), which is of dubious practical value, the more conventional approach of using formalin-inactivated whole-cell preparations (Kusuda and
Fukuda 1980 ; Fukuda and Kusuda 1981b ; Afonso et al. 2005 ) and the more modern
approaches of genetic engineering. Of relevance, the salinity of the growth medium
composition appears to have an effect on the subsequent immune response after
vaccination, with 2.5 % (w/v) rather than 0.5 % NaCl being the more effective
(Nitzan et al. 2004 ). Bacteriological media containing peptones, yeast extract and
salt led to the synthesis of a wider range of cellular components, (including novel
compounds of ~14 and ~21.3 kDa) than those produced in more in vivo type conditions. These compounds were recognised by post-disease sea bass serum
(Bakopoulos et al. 2003b ). It was demonstrated that administration of a formalininactivated preparation in Freund’s complete adjuvant by i.p. injection induced
agglutinating antibodies in yellowtail. Thus titres of 1:256–1:2048 were achieved 5
weeks after vaccination (Kusuda and Fukuda 1980 ). Vaccination enhances the nitric
oxide response, i.e. the production of reactive nitrogen intermediates with their antimicrobial activities, to infection with the pathogen, and is correlated with the level
of protection (Acosta et al. 2005 ). Further work, using a variety of vaccines and
application methods, demonstrated conclusively that fi sh could be protected against
subsequent infection by Ph. damselae subsp . piscicida, although this has been
refuted by some workers (e.g. Hamaguchi and Kusuda 1989 ). Toxoid enriched
whole cells applied by immersion led to a low antibody response an RPS of 37–41 %
in sea bream (Magariños et al. 1994c ). An improved RPS of >60 % after 35 days
resulted from use of an LPS mixed chloroform-killed whole cell vaccine (Kawakami
et al. 1997 ). Using formalin-inactivated cells with or without FCA and a range of
application methods, namely i.p. injection, 5–7 s spray, hyperosmotic infi ltration
and oral uptake via food, Fukuda and Kusuda ( 1981b ) reported encouraging results
within 21 days following artifi cial challenge with Ph. damselae subsp . piscicida.
The best results, conferring 100 % protection to the fi sh, were obtained by use of i.p.
injection or by spraying. The titre of agglutinating antibodies was measured at
between 1:4 and 1:128. A subsequent study by these authors has pointed to the value
of vaccinating with sub-cellular components, notably bacterial LPS (Fukuda and
Kusuda 1982 ). In this connection, a whole cell vaccine in combination with ECPs
was used more successfully than a commercial product by immersion for 1 h and
i.p. injection in sea bass (Bakopoulos et al. 2003a ). However, formalin-inactivated
whole cells administered intraperitoneally achieved an RPS of 96 % in sea bream
(Hanif et al. 2005 ). The question about the nature of the immune response after i.p.
vaccination with or without a booster after 4 weeks with a FIA adjuvanted inactivated whole cell vaccine was addressed by Arijo et al. ( 2004 ), who demonstrated a
Photobacterium damselae subsp. piscicida (= Pasteurella piscicida)