Enrichment in methanol-containing broth — A simple method for the isolation of Saccharomyces from grapes

In this study a simple and effective method was developed for the isolation of Saccharomyces strains from grapes. Aseptically collected grape samples were processed by enrichment in a nutritive basal medium supplemented with 10% (v/v) methanol followed by isolation of yeast strains. Sixteen of the 18 grape samples yielded Saccharomyces strain(s). More than 70% of the isolates belonged to the genus Saccharomyces. Based on phenotype and electrophoretic karyotyping, all strains of Saccharomyces were identified as S. cerevisiae. For several grape samples, varying physiological characters, the number of spores per asci, and the observed chromosome length polymorphisms provided evidence for diversity of S. cerevisiae strains obtained by this enrichment in methanol-containing broth. Results indicated that enrichment in methanol-containing broth is an effective alternative method to facilitate isolation of Saccharomyces strains from grapes. The enrichment method described in this work provides a simple and effective tool for isolation of Saccharomyces strains from grapes. The method may be applied in studying wine fermentation ecology, as well as for the isolation of potential starter strains from grapes

Effect of lactic acid bacteria inoculation on the aflatoxin B1 contamination and the diversity of yeast communities in Aspergillus flavus-contaminated experimental corn silage

The present work aimed to study the yeast communities of whole crop corn silages (CS) that were previously contaminated with aflatoxin-producing Aspergillus flavus (CSCA). In addition, the effect of lactic acid bacterium (LAB) inoculation on the aflatoxin B1 (AFB1) content, genotoxicity, yeast load, and diversity of yeast communities were also investigated. In A. flavus contaminated silages, after two months, the AFB1 content was 40% lower with LAB inoculation, also a lower level of genotoxicity was determined. The number of yeasts cultured from the initial mixture of chopped whole crop corn was 4.8 × 10 7 CFU g −1 wet mass, while only 2.4 × 10 6 CFU g −1 from the CSCA and 7.1 × 10 5 CFU g −1 from the LAB-inoculated CSCA could be cultured. Based on 144 randomly isolated strains, the yeast community of the initial mixture consisted of 8 species. In contrast, the yeast community of CSCA consisted only of 4 species determined by 132 randomly selected isolates. LAB-inoculated CSCA consisted also of 4 species based on 158 randomly isolated strains. Saccharomyces cerevisiae and Pichia kudriavzevii proved to be predominant in the CSCA, while S. cerevisiae and Meyerozyma guilliermondii were the most abundant in the LAB-inoculated CSCA. The species richness was also confirmed by alpha diversity values (1.827, 1.188, and 1.123 as Shannon's indices for CS, CSCA, and LAB-inoculated CSCA, respectively). In response to LAB inoculation, the species diversity decreased considerably