In each experiment, fish in challenge tanks received doses of
F. columnare cells from the same incubated stock solution. In Experiment
1, the bacterial suspension equivalent was 1.2 × 108 colony
forming units per mL (CFUs/mL) and for Experiment 2, the bacterial
suspension equivalent was 6.0 × 107. In brief, F. columnare isolate LV-
359-01 (a genomovar II isolate; Beck et al., 2012) was streaked on
Ordals' Medium (Anacker and Ordal, 1959). After 48 h, the isolate was
dislodged from the agar using a sterile cotton swab and inoculated
into 5 mL of F. columnare Growth Medium (FCGM; Farmer, 2004). This
suspension was incubated at 28 °C for 24 h and was used to inoculate
1 L of FCGM. The inoculated 1-L broth was incubated for up to 24 h at
28 °C in an orbital-shaker incubator set at 200 rpm; when the bacterial
growth reached an absorbance of 0.85 at 550nm, the flaskwas removed
and placed on a stir plate at room temperature
In each experiment, fish in challenge tanks received doses ofF. columnare cells from the same incubated stock solution. In Experiment1, the bacterial suspension equivalent was 1.2 × 108 colonyforming units per mL (CFUs/mL) and for Experiment 2, the bacterialsuspension equivalent was 6.0 × 107. In brief, F. columnare isolate LV-359-01 (a genomovar II isolate; Beck et al., 2012) was streaked onOrdals' Medium (Anacker and Ordal, 1959). After 48 h, the isolate wasdislodged from the agar using a sterile cotton swab and inoculatedinto 5 mL of F. columnare Growth Medium (FCGM; Farmer, 2004). Thissuspension was incubated at 28 °C for 24 h and was used to inoculate1 L of FCGM. The inoculated 1-L broth was incubated for up to 24 h at28 °C in an orbital-shaker incubator set at 200 rpm; when the bacterialgrowth reached an absorbance of 0.85 at 550nm, the flaskwas removedand placed on a stir plate at room temperature
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