title: Combination of culture-independent and culture-dependent molecular methods for the determination of bacterial community of iru, a fermented Parkia biglobosa seeds.
abstract: In this study, bacterial composition of iru produced by natural, uncontrolled fermentation of Parkia biglobosa seeds was assessed using culture-independent method in combination with culture-based genotypic typing techniques. PCR-denaturing gradient gel electrophoresis (DGGE) revealed similarity in DNA fragments with the two DNA extraction methods used and confirmed bacterial diversity in the 16 iru samples from different production regions. DNA sequencing of the highly variable V3 region of the 16S rRNA genes obtained from PCR-DGGE identified species related to Bacillus subtilis as consistent bacterial species in the fermented samples, while other major bands were identified as close relatives of Staphylococcus vitulinus, Morganella morganii, B. thuringiensis, S. saprophyticus, Tetragenococcus halophilus, Ureibacillus thermosphaericus, Brevibacillus parabrevis, Salinicoccus jeotgali, Brevibacterium sp. and uncultured bacteria clones. Bacillus species were cultured as potential starter cultures and clonal relationship of different isolates determined using amplified ribosomal DNA restriction analysis (ARDRA) combined with 16S-23S rRNA gene internal transcribed spacer (ITS) PCR amplification, restriction analysis (ITS-PCR-RFLP), and randomly amplified polymorphic DNA (RAPD-PCR). This further discriminated B. subtilis and its variants from food-borne pathogens such as B. cereus and suggested the need for development of controlled fermentation processes and good manufacturing practices (GMP) for iru production to achieve product consistency, safety quality, and improved shelf life.
authors: Department of Microbiology, Faculty of Science, University of Lagos Akoka, Lagos, Nigeria ; Microbial Resources Division, Institute of Bioresources and Sustainable Development, Takyelpat Institutional Area Imphal, Manipur, India ; Department of Food Science and Technology, College of Food Sciences, Bells University of Technology Ota, Nigeria.
authors: Gbenga A
authors: GA
authors: Adewumi
authors: Folarin A
authors: FA
authors: Oguntoyinbo
authors: Santosh
authors: S
authors: Keisam
authors: Wahengbam
authors: W
authors: Romi
authors: Kumaraswamy
authors: K
authors: Jeyaram
keywords: Bacillus; DGGE; fermentation; iru; safety