The reason for differences in the virulence between different isolates
of A. hydrophila is due to a wide variation in the expression of
genes between various isolates, which in turn leads to different levels
of expression of the virulence factors, such as those found in the
ECP or as surface proteins [37]. In this study, the lowest virulence
was seen with isolate T4 and the highest with isolate B2/12. The rate
of mortality was high with all six isolates in both vaccinated and
control fish within the first 2 days post-challenge, compared with
the level of mortality obtained over the rest of the trial (Fig. 4). The
sudden mortality that occurred in the first 2 days post-challenge
was most likely due to toxic shock [38]. This rate of mortality is
unlikely to occur during a natural infection because the concentration
of the pathogen gradually increases during the infection,
whereas a large number of bacteria are introduced at the same time
in the experimental infection. The recombinant S-layer protein vaccine
may therefore have a greater ability to protect fish against
natural infections by A. hydrophila, when bacterial concentrations
are low.
The S-layer protein is a predominant cell surface protein seen
in the SDS-PAGE profiles of WC lysates and outer membrane fractions
of A. hydrophila [39]. The presence of S-layer protein among
highly virulent strains of A. hydrophila has previously been reported
by Thomas and Trust [32] and Dooley et al. [40]. Diseases caused
by A. hydrophila possessing S-layers are often associated with invasive
systemic infection [41]. Being on the outermost layer of the
bacterium, the S-layer protein has more chance of rapidly interacting
with the host than other protein components of the bacterium
[32]. The S-layer binds to many host proteins such as fibronectin,
laminin and vitronectin [42], which could be one reason why the
S-layer protein appears to be more immunogenic than other proteins
in the bacterium. Kokka et al. [43] suggested that the S-layers
may provide protection for bacteria in their natural environment
or provide a selective advantage in the ability of bacterium to cause
infection. The protein was also found to confer resistance to serum
killing and protease digestion [42].
The study indicated that the S-