Distribution of Aspergillus section Nigri at shochu fermenting places in Japan

Koji mold, which belongs to the Aspergillus section Nigri, is used in the production of shochu. The section Nigri is composed of very morphologically similar members that in some cases produce mycotoxins, which rises concerns as to whether the presence of mycotoxin-producing fungi in shochu producing sites can compromise consumer safety. Thus, we examined the presence of mycotoxin-producing sec. Nigri fungi in six shochu factories (named A-F) in Japan. Airborne fungal levels in the factories were determined, and a traditional koji called "kona-koji" made from the mold naturally present in factory C (Aogashima village) was analyzed. Isolates of sec. Nigri fungi were identified morphologically and confirmed via cytochrome b gene analysis. In factory A (Nago city), airborne fungal levels of sec. Nigri were 4,000 and 100 cfu/m³ in the koji-making and fermentation rooms, respectively. In factories B, C, and D, the levels were 40, >10⁴ cfu/m³, and 100 cfu/m³, respectively. In factory F (Iki city), there were high levels of airborne white-koji mold (a white mutant of Asp. luchuensis). The most dominant fungal species of sec. Nigri was isolated and identified as Asp. luchuensis via genetic analysis. This is likely to have originated from the commercial fermentation culture used. Asp. niger and Asp. luchuensis were isolated from kona-koji. Mycotoxin production (ochratoxin and fumonisin B2) by Asp. luchuensis (eight strains) and Asp. niger (three strains) was virtually inexistent; only one strain of Asp. niger was positive for fumonisin B₂. This study clearly shows that mycotoxin-producing fungi are not dominant in the fungal flora present in the shochu factories examined and therefore, that the liquor can be safely fermented.Implications: In this study, we examined the presence of mycotoxin-producing Aspergillus sec. Nigri fungi in six shochu (Japanese distilled beverage) factories. The most dominant fungal species of sec. Nigri was isolated and identified as Aspergillus luchuensis (black-koji mold). The proportion of mycotoxin-producing Aspergillus niger and Aspergillus carbonarius was very small. In addition, the Asp. niger isolated from koji mold did not have the ability to produce ochratoxins or fumonisin B. This study clearly shows that shochu can be safely fermented.