Protein expression by Aeromonas hydrophila during growth in vitro and in vivo

Expression of Aeromonas hydrophila cellular and extracellular products (ECPs) was examined following culture of the bacterium in vitro, in Tryptic Soy Broth (TSB), and in vivo, in dialysis tubing placed within the peritoneal cavity of common carp (Cyprinus carpio L.). Whole cell (WC), outer membrane proteins (OMPs) and ECP components of the bacteria were analysed by 1 dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (1D SDS-PAGE). Additionally, 2D SDS-PAGE was used to analyse WC preparations. The aim of the study was to identify unique and common proteins up-regulated in vivo. Unique bands were seen in the 1D gels at 58 and 55 kDa for WC and OMP preparations, respectively, for all the four virulent and two avirulent isolates cultured in vivo. Bands of increased intensity were also observed at 70, 55, 50 and 25 kDa with WC preparations for all virulent isolates cultured in vivo. Analysis of WC by 2D SDS-PAGE revealed that bacteria cultured in vivo expressed a number of unique spots, mostly between 30 and 80 kDa with pI values ranging from 5.0 to 6.0. The unique proteins identified in vivo may be involved in the virulence of the bacterium and their potential as vaccine candidates is currently being investigated.