Evaluation of media for enumerating heat-stressed, benzoate-resistant Zygosaccharomyces bailii

A study was undertaken to determine the efficacy of yeast malt extract agar (YMA), acidified YMA (YMAA), acidified tryptone glucose yeast extract agar (TGYA) and Zygosaccharomyces bailii agar (ZBA) for enumerating two strains of Z. bailii (FRR 2227 and FRR 3680) grown and heated in blueberry syrup containing 0, 300 and 600 micrograms/ml sodium benzoate. Cells were heated at 50, 51 or 52 degrees C for 0, 15, 30 or 45 min before spread plating diluted samples on enumeration media and incubating at 30 degrees C for 5 days. Strain FRR 3680 exhibited higher heat resistance than did strain FRR 2227. Regardless of the sodium benzoate concentration in growth and heating media, or the temperature and time of heating, non-selective YMA supported recovery of the highest number of viable cells, followed by TGYA for strain FRR 2227 and TGYA or ZBA for strain FRR 3680. YMAA supported the lowest recovery of heat-stressed cells. The concentration of sodium benzoate in the growth medium did not have a significant effect on tolerance of strain FRR 3680 to heat; however, tolerance of strain FRR 2227 to heat decreased when cells were grown in blueberry syrup containing increased concentrations of sodium benzoate. Regardless of the sodium benzoate concentration in the growth medium, both strains exhibited increased sensitivity when heated in blueberry syrup containing increased concentrations of sodium benzoate. Overall, TGYA was judged best among selective media for enumerating Z. bailii because of its ability to support colony development by heat-stressed cells and because of the ease of counting and differentiating colonies.