FIGURE 2.

Schematic showing the different steps examined in the variable temperature growth and exposure experiments. Three different treatment protocols are indicated in the figure. Route (A) provides a control experiment for samples exposed to routes (B,C). In route (A) cultures that had previous experienced only ambient pressure and temperature conditions were grown at 30°C while being rotated at 180 rpm. Once a pellet was obtained the sample was plated and aliquots were incubated at the different temperatures investigated (i.e., 37, 30, 25, 19, and 8°C). Colonies formed were counted and the results are shown in Table 2. Route (B) cultures were obtained from survivors of the wild type (WT) strain that had been exposed to 1000 MPa ( Table 1). These were cultured at 30°C and 180 rpm from stocks created from the survivor population. Each sample was then incubated at different temperatures (37, 30, 25, 19, and 8°C). Route (C) cultures were likewise isolated from survivors of the initial 1000 MPa exposure experiment, cultured at 30°C and 180 rpm from stocks. In the next step these samples were exposed to different temperatures (30, 25, 19, or 8°C) while being held once more at high pressure (1000 MPa).