International Journal of Applied Microbiology and Biotechnology Research
ISSN: 2053-1818
Vol. 5(9), pp. 95-102, October, 2017



Bacterial diversity study on fermentation of Parkia biglobosa using 16s rRNA gene analysis

Aminat Olabisi Adelekan1*, Sylvia V. A. Uzochukwu2 and Nwadiuto Esiobu3

1Department of Chemical and Food Sciences, Bells University of Technology, P. M. B. 1015, Ota, Ogun State, Nigeria.
2Department of Plant Science and Biotechnology, Federal University, Oye-Ekiti, Ekiti State, Nigeria.
3Biological Sciences Department, Florida Atlantic University, 3200 College Avenue, Davie Florida 33314, U.S.A.

*To whom correspondence should be addressed. E-mail: bis_adek@yahoo.com.

Received 25 April, 2015; Received in revised form 17 June, 2015; Accepted 02 September, 2017.

Abstract


Keywords:
Bacterial diversity, Parkia biglobosa, 16S rRNA, Molecular methods.

The diversity and dynamics of the bacterial community during the fermentation of Parkia biglobosa to produce ‘Iru’, a condiment, were studied, using 16S rRNA gene analysis. Fresh P. biglobosa seeds were processed using the traditional method at ambient temperature (30±2°C) for 72 h. The total bacterial community was obtained by vigorously rinsing the seeds in phosphate-buffered saline at 0 h and subsequently at 24, 48 and 72 h. Community DNA was extracted directly from the rinsed water and from nutrient culture of the rinse water. The 16S rRNA genes of bacteria present were amplified by polymerase chain reaction from the extracted DNA, using the primer pair 27F and 1492R. The amplicons were sequenced and aligned with reference fragments in the database of the ribosomal RNA project. The bacterial populations in fermented ‘Iru’ increased from 1.2x101 cfu/g at start of fermentation to 1.63 × 109 cfu/g at 72 h. The results obtained from the database were as follows: Bacilli–Bacillales represented by Bacillus algicola; Bacillus cereus; Bacillus thuringiensis; Bacillus subtilis; Bacillus pumilus; Bacillus sp.; uncultured Staphylococcus sp.; Bacillus anthracis; Bacillus foraminis; Bacillus clausii; Lysinibacillus spp.; Lactobacillales was represented by Streptococcus sanguinis; Fusibacter sp.; Clostridia–Clostridales was represented by Butyrivibrio fibrisolvens; Bacteroidia –Bacteriodales was represented by Bacteroides nordii; Parabacteroides merdae; Alpha proteobacter was represented by Acetobacter pasteurianus; gamma proteobacter represented by Acinetobacter baumannii and Enterobacter aerogenes. The dominance of Bacillus spp. in cultured bacterial community increased from 8 to 83%, while that of alpha proteobacteria increased from 2 to 15% and gamma proteobacteria decreased from 28 to 1%. As fermentation progressed Bacillus species decreased. Gamma proteobacteria increased to 17% at 72 h. The succession of the bacteria associated with the fermentation of P. biglobosa to ‘Iru’ was shown to be co-dominated by Bacillus and the Proteobacteria spp. Molecular method adopted showed varieties of microorganisms that had hitherto not been identified by the conventional culture-dependent methods.

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