Preliminary studies of the effects of heat and gamma irradiation on the production of aflatoxin B 1 in static liquid culture, by Aspergillus flavus link NRRL 5906

Abstract

Aflatoxin B 1 production by a strain of Aspergillus flavus NRRL 5906 was examined in static liquid culture in maize meal broth (MMB) and maize meal broth supplemented with 2% glucose and 2% peptone (AMMB). Erlenmeyer flasks were inoculated with 1.0 ml aliquots of fungal spores which had been heat-treated (60°C for 30 min) under low humidity (< 45% R.H. dry heat) or high humidity conditions (>85% R.H., moist heat) followed by gamma irradiation with either 0.0, 3.5 or 4.0 kGy. AMMB supported 6–17 times more vegetative growth (depending on the heat and dose combination) than spores incubated in MMB alone. High inoculum size of control unheated spores (log CFU/g, 6.9) yielded the least aflatoxin B 1 in flasks containing AMMB (8.2–19.3 μg/ml). A dose of 3.5 kGy reduced by 3.2–3.8 log cycles the viable inoculum of control unheated spores, resulting in 2–5 fold increase in aflatoxin B 1 formed in flasks containing AMMB. Increasing the applied load to 4.0 kGy, however, reduced aflatoxin B 1 levels formed in AMMB to similar or lower levels than found in flasks inoculated with control unirradiated spores. Combination treatment of A. flavus with dry heat and 3.5 kGy reduced the spore inoculum size by about 4 log cycles and yielded the highest amount (41.1 μg/ml) of aflatoxin B 1 in AMMB. However, moist heat treatment of spores receiving the same dose (3.5 kGy) reduced toxin level formed by 25%. Aflatoxin B 1 formation by A. flavus spores incubated in AMMB was completely prevented by a combination treatment of moist heat and 4.0 kGy of gamma irradiation. This same treatment attenuated aflatoxins B 2, G 1 and G 2 production which are formed with B 1 by A. flavus NRRL 5906. Spores raised in all flasks containing MMB did not form aflatoxin except when the medium MMB was autoclaved twice at 121°C for 15 min.