United Kingdom. Bacteria were inoculated into 10 ml of liquid tryptic soy broth
(TSB, Sigma) medium and were grown overnight at 28 °C. Cultures were
centrifuged at 850 G for 15 min. Supernatant was removed and the pelleted
bacteria were washed twice in sterile phosphate buffered saline (PBS) solution.
The concentration of bacteria was adjusted to 5×107 by the optical density of
suspension.
0.1 ml of suspended bacteria was injected into the peritoneal cavity of fish.
Mortality was recorded during the ten days following infection.