Antagonism of nonaflatoxigenic Aspergillus flavus isolated from peanuts against aflatoxigenic A. flavus growth and aflatoxin B1 production in vitro.

Aspergillus section Flavi constitutes several species of opportunistic fungi, notable among them are A. flavus and A. parasiticus, capable of surviving harsh conditions and colonizing a wide range of agricultural products pre- and postharvest. Physical and chemical control methods are widely applied in order to mitigate the invasion of A. flavus in crops. However, physical control is not suitable for large scale and chemical control often leads to environmental pollution, whereas biological control offers a safer, environmentally friendly, and economical alternative. The present study aimed to investigate the antagonism of several non-aflatoxigenic A. flavus strains against the aflatoxigenic ones in vitro (semisynthetic peanut growth medium; MPA) in terms of colony growth rate and AFB1 inhibition. Different peanut concentrations were used to obtain the optimum peanut concentration in the formulated growth medium. A dual culture assay was performed to assess the antagonism of nonaflatoxigenic strains against the aflatoxigenic ones. Results revealed that 9% MPA exhibited the highest growth and AFB1 inhibition by nonaflatoxigenic strains. It was also found that different nonaflatoxigenic strains exhibited different antagonism against the aflatoxigenic ones which ranged from 11.09 ± 0.65% to 14.06 ± 0.14% for growth inhibition, and 53.97 ± 2.46% to 72.64 ± 4.54% for AFB1 inhibition. This variability could be due to the difference in antagonistic metabolites produced by different nonaflatoxigenic strains assessed in the present study. Metabolomics study to ascertain the specific metabolites that conferred the growth and aflatoxin inhibition is ongoing.

Prevalence, Identification and Mycotoxigenic Potential of Fungi in Common Spices Used in Local Malaysian Cuisines.

Spices are widely used in various cuisines in Malaysia to enhance the flavour and aroma. However, spices are susceptible to fungal infection, leading to mycotoxin contamination if the storage conditions are favourable for fungal growth. Thus, this study aimed to identify fungal species in spices commonly used in local Malaysian cuisines and determine their prevalence and mycotoxigenic potential. A total of 110 spice samples consisting of cumin, fennel, coriander, peppers (black pepper and white pepper), chillies (dried chilli, chilli paste and chilli powder), cinnamon, star anise, cloves, curry powder and korma powder were randomly purchased from retail markets in Penang. The samples were analysed for the total fungal count (ground spices) and the incidence of fungal infection (whole spices). The fungal species isolated from spices were identified based on morphological and molecular approaches, and the mycotoxigenic potential was determined using the Coconut Cream Agar method. The results showed that coriander seeds (ground) recorded the highest total fungal count (ADM 3.08 log CFU/g; DG18 3.14 log CFU/g), while black pepper (whole) recorded the highest incidence of fungal infection (94%). Interestingly, star anise and cloves were free from fungal contamination. The mycotoxigenic fungi of A. flavus and A. niger recorded the highest isolation frequency in ground and whole spices. These findings indicate the risk of mycotoxin exposure to consumers due to the high consumption of spices in local Malaysian cuisine.

Formulation of maize- and peanut-based semi-synthetic growth media for the ecophysiological studies of aflatoxigenic Aspergillus flavus in maize and peanut agro-ecosystems.

In studying the ecophysiology of fungal phytopathogens, several stages are involved (in vitro, greenhouse, in planta). Most in vitro studies extensively utilise the general growth media such as Potato Dextrose Agar and Malt Extract Agar. Although the crop components in these media serve as excellent carbon sources and yield luxuriant growth, they are not naturally contaminated with Aspergillus flavus and thus might result in under- or overestimation of its actual toxigenic potentials. Empirical data on the formulation of semi-synthetic growth medium mimicking the natural crop commonly contaminated by A. flavus for the ecophysiological studies in vitro are scarce. The present work was aimed at investigating the ecophysiology of A. flavus on commercial growth media (PDA, MEA); formulating maize- and peanut-based semi-synthetic growth media using two methods of raw material preparation (milling, hot water extraction) at different concentrations (1, 3, 5, 7, 9% w/v), and comparing the ecophysiological parameters between commercial and formulated growth media. Growth rates were obtained by computing the hyphal expansion data into y = mx + c equation. AFB1 was quantified using high performance liquid chromatography with fluorescence detector. Formulated media were found to yield significantly higher growth rates when compared to commercial media. However, commercial media yielded significantly higher AFB1 when compared to all formulated media. Between the two formulations, milling yielded significantly higher growth rates and AFB1 when compared to hot water extraction. Although in vitro data cannot directly extrapolate in planta performance, results obtained in the present work can be used to gauge the actual toxigenic potential of A. flavus in maize and peanut agro-ecosystems.