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.
Highlights
Fermented vegetable protein seeds used for condiments production in West Africa include African locust bean [Parkia biglobosa
Representative of the major clusters obtained were identified by sequence data of 16S rRNA gene that identified two major bacilli groups; B. subtilis and B. amyloliquefaciens as the dominant, as well as B. cereus group as member of the cultivatable bacteria associated with fermented P. biglobosa cotyledon (Table 1)
ANALYSIS OF BACTERIAL COMMUNITIES AND DIVERSITY OF iru USING PCR-denaturing gradient gel electrophoresis (DGGE) The two genomic DNA extraction protocols- enzymatic and chemical proved to be efficient in terms of quantity, purity, and amplifiabiltiy, and produced DNA suitable for PCR amplification
Summary
Fermented vegetable protein seeds used for condiments production in West Africa include African locust bean [Parkia biglobosa Condiments constitute a significant proportion of African diets where they serve as flavorsome and culinary components in various dishes (Achi, 2005) They include iru or dawadawa in Nigeria and Ghana; soumbala, bikalga, and maari in Burkina Faso; afitin and sonru in Benin Republic; nététou in Senegal; kinda in Sierra Leone; dawadawa botso in Niger Republic; datou in Mali; mbuja in Cameroon and furundu in Sudan (N’Dir et al, 1994; Ouoba et al, 2008, 2010; Parkouda et al, 2010). The art is carried out in exclusive uncontrollable environmental conditions thereby yielding products with variation in quality and organoleptic properties (Sanni, 1993; Steinkraus, 1997; Ouoba et al, 2007) This necessitates the need for the development of appropriate starter cultures to initiate fermentation for the production of consistent products with acceptable qualities
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