Inhibitory Effects of Eurotium cristatum on Growth and Aflatoxin B(1) Biosynthesis in Aspergillus flavus

Probiotic strain Eurotium cristatum was isolated from Chinese Fuzhuan brick-tea and tested for its in vitro activity against aflatoxigenic Aspergillus flavus. Results indicated that E. cristatum can inhibit the radial growth of A. flavus. Furthermore, this inhibition might be caused by E. cristatum secondary metabolites. The ability of culture filtrate of strain E. cristatum against growth and aflatoxin B(1) production by toxigenic A. flavus was evaluated in vitro. Meanwhile, the influence of filtrate on spore morphology of A. flavus was analyzed by scanning electron microscopy (SEM). Results demonstrated that both radial growth of A. flavus and aflatoxin B(1) production were significantly weakened following increases in the E. cristatum culture filtrate concentration. In addition, SEM showed that the culture filtrate seriously damaged hyphae morphology. Gas chromatography mass spectrometry (GC/MS) analysis of the E. cristatum culture supernatant revealed the presence of multiple antifungal compounds. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis showed that the expression of aflatoxin biosynthesis-related genes (aflD, aflQ, and aflS) were down-regulated. Importantly, this latter occurrence resulted in a reduction of the AflS/AflR ratio. Interestingly, cell-free supernatants of E. cristatum facilitated the effective degradation of aflatoxin B(1). In addition, two degradation products of aflatoxin B(1) lacking the toxic and carcinogenic lactone ring were identified. A toxicity study on the HepG2 cells showed that the degradation compounds were less toxic when compared with AFB(1).