||Two Aeromonas hydrophila strains, ATCC 7966' and a food
isolate, JAH 4 (Hudson 1992b), were grown on
Brain Heart Infusion broth (Difco, Detroit, MI,
USA), containing 10 g/liter 3-[N-morpholino]
propane sulphonic acid (Sigma Chemical Co., St
Louis, MO, USA) as a buffer, adjusted to pH
7.0 before autoclaving. Inocula were grown at
either 5, 15, 25 or 35°C in shaking water baths
and subcultured into pre-equilibrated media at
these four temperatures (16 combinations).
Inocula were subcultured three times at the preincubation
temperature, when the optical
density (O.D.) at 650 nm was 0.5 ensuring that
subcultures were made during the early
logarithmic phase of growth.