Modified Pikovskaya medium was used for all experiments
unless otherwise stated, containing (g L−1 in distilled water) glucose
10.0, (NH4)2SO4 0.5, NaCl 0.2, MgSO4·7H2O 0.1, KCl 0.2, yeast
extract 0.1, MnSO4 trace amount, FeSO4·7H2O trace amount. The
pH was adjusted to 6.8 for all experiments before autoclaving.
Phosphate (PO4
3−) was added at 3.0 g L−1 as tri-calcium phosphate
Ca3(PO4)2. Flasks were inoculated 105 (±5 × 105) cells of either
CtHY or the reference strain, Klebsiella pneumoniae 4P (Ahmed,
2006). All experiments were conducted with triplicate sample
flasks for each time sample, with each flask containing 25 mL of
liquid growth media. Flasks were incubated aerobically with shaking
(100 rpm) for 10 days at 30 ◦C. At each sample time 100 L of
culture suspension was taken and serially diluted for plate counting.
The remainder of the sample was centrifuged at 5000 × g for
15 min and the pH and soluble P in the supernatant were analysed.
The pH was measured with a PHM210 meter (Radiometer Analytical,
France). Soluble P was measured using the ascorbic acid method
(Olsen et al., 1954).
Modified Pikovskaya medium was used for all experimentsunless otherwise stated, containing (g L−1 in distilled water) glucose10.0, (NH4)2SO4 0.5, NaCl 0.2, MgSO4·7H2O 0.1, KCl 0.2, yeastextract 0.1, MnSO4 trace amount, FeSO4·7H2O trace amount. ThepH was adjusted to 6.8 for all experiments before autoclaving.Phosphate (PO43−) was added at 3.0 g L−1 as tri-calcium phosphateCa3(PO4)2. Flasks were inoculated 105 (±5 × 105) cells of eitherCtHY or the reference strain, Klebsiella pneumoniae 4P (Ahmed,2006). All experiments were conducted with triplicate sampleflasks for each time sample, with each flask containing 25 mL ofliquid growth media. Flasks were incubated aerobically with shaking(100 rpm) for 10 days at 30 ◦C. At each sample time 100 L ofculture suspension was taken and serially diluted for plate counting.The remainder of the sample was centrifuged at 5000 × g for15 min and the pH and soluble P in the supernatant were analysed.The pH was measured with a PHM210 meter (Radiometer Analytical,France). Soluble P was measured using the ascorbic acid method(Olsen et al., 1954).
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