Isolation and Identification of Lactic Acid Bacteria from Spontaneously Fermented Kunun-zaki using RAPD-PCR Analysis
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Abstract
Sixty lactic acid bacteria (LAB) were isolated from eighteen types of kunun-zaki produced naturally using millet, sorghum and maize each singly or in combinations with malted grains (millet, sorghum, maize, wheat and paddy rice) and characterized using conventional methods (biochemical and cultural) and molecular analyses. Seven morphotypes (Mt1-Mt7) were revealed among the 60 LAB isolates. Twenty LAB were selected based on the differences in their biochemical and physiological properties and these were subjected to DNA isolation and purification. The Agarose gel electrophoresis of the twenty (20) extracted DNA revealed very pure DNA electrophoretic pattern. The concentration of the DNA sample ranged between 0.7 mg/ml to 4.0 mg/ml and the purity index ranged from 1.5 to 1.9; indicating they were all very pure and can easily be used in Polymerase chain reaction (PCR) analysis. A highly polymorphic primer, OPB20 primer, was selected and used to analyze the 20 LAB DNA of which seven (7) isolates were identified to be Lactobacillus brevis, twelve (12) isolates were Lactobacillus plantarum and one isolate was identified as Lactobacillus acidophilus and these organisms could be potentially useful industrially.
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References
Abiodun, O.A., Dauda, A.O., Adebisi, T.T. and Alonge, C.D. (2017).Physico-cchemical, microbial and sensory properties of kunun-zaki beverage sweetened with black velvet tamarind (Dialium guineese).Croat. J. Food Sci.Technol.9 (1):07
Agarry ,O. O., Nkama, I and Akoma, O. (2010). Production of kunun- zaki (A Nigerian fermented cereal beverage) using starter culture. International Research Journal of Microbiology, 1 (2):18-25.
Ajayi, O. D., Onasanya, A.A.,Ojo, A. A., Obafemi, T.O. and Daramola, G.O. (2023) Distribution of lasl and rhlR virulent gene among Staphylococcus aureus isolates in clinical samples from Ekiti State University Teaching Hospital, Ado-Ekiti, Nigeria. GSC Biological and Pharmaceutical Sciences, 22 (03), 049-081
Ayandele, A. A. (2015). Microbiological analyses of hawked kunun and zobo Drinks within LAUTECH campus, Ogbomoso, Oyo-state. Journal of Environmental Science, Toxicology and Food Technology 9(10):52-56
Beetseh, C.I., Yoo, M.A. and Achikpi, F. (2022)Effects of different preservative treatment on the sugar content of kunun-zaki in University of Agriculture, Makurdi,
Benue State,Nigeria. Journal of Environment and Earth Science, ISSN2224-3216 (Paper),Online,vol.12 (2).
Bintsis, T. (2018). Lactic acid bacteria as starter cultures: An update in their metabolism and genetics. AIMS Microbiology, 4(4):665-684.
Helmenstine , A.M.(2019). Gram StainProcedure in Microbiology. ThoughtCo.thoughtco.com
Kiran,P.(2018) Surprising Benefits of Cloves. Organic Facts. https://www.organic facts.net/health-benefits/herbs and-spices/heal...:1-6
Kyule, D.N.,Maingi, J. M., Njeru, E.M. and Nyamache, A.K.(2022) Molecular characterization and diversity of bacteria isolated from fish and fish products retailed in Kenyan markets, International Journal of Food Science, volume 2022,Article ID 2379323, 12pages, https//doi.org/10.1155/2022/2379323.
Matsumoto, S., Watanabe, K.,Kiyota, H., Tachibata, M.,Shimizu, T. and Watarai, M.(2022) Distinction of Paramecium strains by a combination method of RAPD analysis and multiplex PCR
Michael, J. (2019) Amazing Health Benefits of Cloves. Natural Food Series, https://www.natural foodseries.com> Food and Nutrition:1-8.
Mulaw, G., Tefaye, S.T., Diriba,, M. and Anteneh, T. (2019) In vitro evaluation of probiotic properties of lactic acid bacteria isolated from some traditionally fermented Ethiopian food products. https://doi.org/10./155/2019/7179514
Ojo, A.,Olawale, A., Okiki, P., Oyinloye,F.,Ogunnusi, T., Onasanya, A. and Morakinyo, O (2019). Virulence characteristics and phylogenetic background of antibiotic-resistant Enterococcus faecalis from abattoir, poultry and clinical origin in Ado-Ekiti, Nigeria. African Journal of Microbiology Research, vol. 13(30), pp.700-709.
Onasanya, A. Onasanya, R. O. Ojo, A. A. Adewale, B. O. (2013). “Genetic Analysis and Molecular Identification of Virulence in Xanthomonas oryzae pv. oryzae Isolates", International Scholarly Research Notices, vol. 2013, Article ID 160157, 8 pages, https://doi.org/10.1155/2013/160157
Onyimba, I.A., Itelima, J.U., Job, M.O., Ogbonna, A.I. and Ode, C.O.(2017). Production of Sorghum Based Kunun Zaki Using Selected Starter Cultures. International Journal of Sciences 6 (9):1370
Pulse, Nigeria (2018). Health Benefits of kunun Pulse Nigeria https://www.pulse.ng/lifestyle/food-travel/food-and-health-health:2
Paddock, C. (2019). This mechanism could be why fermented foods benefit immune heath https://www.medicalnewstoday.com
Ray, R.C. and Joshi, V.K. (2014). Fermented Foods: Past, Present and Future. ResearchGate. https://www.researchgate.net>publication.
Rohlf,F.J.(2000). Numerical Taxonomy and Multivariate Analysis System, NTSYS-pc.,Version2.1. Exeter Publishing Setauket, New York.
Sagar, A.(2022) Catalase test-Principle, uses, Procedure, Result Interpretation with Precautions. microbiologyinfo.com
Sagar, A. (2022) .Potassium Hydroxide Test – Principle, Procedure, Uses and Interpretation Potassium Hydroxide Test - Principle, Procedure, Uses and Interpretation (microbiologyinfo.com)
Somashekaraiah, R.,Shruthi, B.,Deppthi, B. V. and Sreenivasa, M. Y. (2019).Probiotic properties of Lactic acid bacteria isolated from Neera: A naturally fermenting coconut palm nectar. Front. Microbiol.,28 https://doi.org/10.3389/fmicb:2019.01382
Thottappily, G., Mignouna, H.D., Onasanya, A.,Abang, M.,Oyelakin, O. and Singh,N.K.(1999). Identification and differentiation of isolates of Colletotrichum gloeosporioides from yam by random amplified polymorphic DNA markers. African Crop Science Journal, vol. 7, No 2, pp.195-205.
Victor-Aduloju A. T., Anyamene C., Ogbu K. N. and Ishiwu C. N.(2018). Isolation and identification of probiotic Lactobacillus species from traditional drink kunun-zaki fortified with paddy rice and sweet potatoes. African Journal of Food Science,12(9): 230-237.