each time point sampled, five replicate capillary
tubeswere prepared for a total of 100 μl volume of cell suspension.
Non-treated controls were included as “time 0” data points. Capillary
tubes containing hyphal phase cells were incubated in water baths at
temperature 54 °C, 56 °C, and 58 °C. Capillary tubes containing yeastlike
cells were incubated in water baths at temperature 51 °C, 53 °C,
and 55 °C. Sampling points were recorded at intervals appropriate to
the rapidity of cell death for a given temperature.
At selected time points, capillary tubes were removed from the
water bath and immediately submerged in 70% ethanol on ice.
Surface-sterilized capillary tubes were blotted, transferred to milk dilution
bottles containing 20 ml peptone water, and crushed using a glass
rod. Subsequent serial dilutions were plated using acidified PDA. Plates
were incubated at 25 °C for 48 h, then enumerated.