Genetic diversity of lactic acid bacteria isolated from Sumbawa horse milk, Indonesia
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http://orcid.org/0000-0001-6614-5518
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Abstract. Kusdianawati, Mustopa AZ, Fatimah, Budiarto BR. 2020. Genetic diversity of lactic acid bacteria isolated from Sumbawa horse milk, Indonesia. Biodiversitas 21: 3225-3233. LAB from Sumbawa horse milk has good potential antimicrobial and probiotic agents. It is known, the study on LAB diversity based on its phenotypic characters is difficult to be distinguished. However, the development of molecular characterization based on the genotypic characteristic could be done for LAB diversity analysis. The aim of this study is to obtain the genetic diversity of LAB from Sumbawa horse milk collected from Penyaring Village and Lennanguar Village, Sumbawa, West Nusa Tenggara Indonesia. LAB strains were identified based on their genotypic characteristics, including their randomly amplified polymorphic DNA (RAPD) primers profiles and 16S ribosomal RNA (rRNA) sequences. The result of RAPD-PCR analysis showed 5 clusters of dendrograms resulted from GTG5 and LB2 primer amplification. Based on 16 rRNA sequences result, the phylogenetic tree was constructed and revealed 7 species of LAB i.e: SK 1.5, SKP K.3, SKP K.5, SKP K.9/SKP K.7/M.SKP K.3, SKL K.4, M.SKL K.1/ M.SKL K.5, and SKP K.4 belonging to the species of Enterococcus faecium, Weissella confusa, Lactococcus garvieae, Enterococcus thailandicus, Lactobacillus fermentum, Enterococcus faecalis, and Lactococcus petauri. In this study, the bacteria from Enterococcus sp., Lactococcus garvieae, and Lactococcus petauri existed as a novel of bacteria which means they have not been isolated and identified in Sumbawa horse milk compared to the previous findings.
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References
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Mikelsaar M, Zilmer M. 2009. Lactobacillus fermentum ME-3 an antimicrobial and antioxidative probiotic. Microb Ecol Health Dis 21(1): 1–27.
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Tenea GN, Lara MI. 2019. Antimicrobial compounds produced by Weissella confusa Cys2-2 strain inhibit Gram-negative bacteria growth. CYTA-J Food 17(1): 105–111.
Villani F, Aponte M, Blaiotta G, Mauriello G, Pepe O, Moschetti G. 2001. Detection and characterization of a bacteriocin, garviecin L1-5, produced by Lactococcus garvieae isolated from raw cow’s milk. J Appl Microbiol 90 (3) : 430–439.
Welsh J, McClelland M. 1990. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18 (24) : 7213-8.
Widiada IGN, Antara NS, Aryanta WR. 2006. Identification and growth succession of lactic acid bacteria in wild horse milk of Bima during storage. In Annual Scientific Meeting of PERMI-2006. Udayana University, Bali. [Indonesia].
Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18 (22) : 6531-5.
Yu J, Wang WH, Menghe BL, Jiri MT, Wang HM, Liu WJ, Bao QH, Lu Q, Zhang JC, Wang F, Xu HY, Sun TS, Zhang HP. 2011. Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia. Int J Dairy Sci 94(7): 3229-41.
Zacharof MP, Lovitt RW. 2012. Bacteriocins produced by lactic acid bacteria. APCBEE Procedia 2: 50 - 56.
Zhu H, Qu F, Zhu LH. 1993. Isolation of genomic DNAs from plants, fungi, and bacteria using benzyl chloride. Nucleic Acids Res 21: 5279-5280.
Bauer R, Bekker JP, Wyk NV, du Toit C, Dicks LMT, Kossmann J. 2009. Exopolysaccharide production by lactose-hydrolyzing bacteria isolated from traditionally fermented milk. Int J Food Microbiol 131(2-3):260–264.
Ben Omar N, Ampe F, Raimbault M, Guyot JP, Tailliez P. 2000. Molecular diversity of lactic acid bacteria from Cassava sour starch. Syst Appl Microbiol 23 : 285-290.
Björkroth JA, Dicks LMTD, Endo A. 2014. The Genus Weissella, in Lactic Acid Bacteria, Biodiversity and Taxonomy. Wiley-Blackwell, Chischester.
Cai, Y, Yang J, Pang H, Kitahara M. 2011. Lactococcus fujiensis sp. nov., a lactic acid bacterium isolated from vegetable matter. Int J Syst Evol Microbiol 61: 1590–1594.
Centeno JA, Cepeda A, Rodriquez-Otero JL. 1995. Identification and preliminary characterization of strains of enterococci and micrococci isolated from Arzúa raw cows'-milk cheese. Nahrung 39 : 55-62.
Cogan TM, Barbosa M, Beuvier E, Bianchi-Salvadoris B, Cocconcelli PS, Fernandes I, Gomez J, Gomez R, Kalantzopoulos G, Ledda A, Medina M, Rea MC, Rodriguez E. 1997. Characterization of the lactic acid bacteria in artisanal dairy products. J Dairy Res 64: 409-421.
Chao SH,Tomii Y,Watanabe K, Tsai YC. 2008. Diversity of lactic acid bacteria in fermented brines used to make stinky tofu. Int J Food Microbiol 123: 134-141.
Chao SH, Huang HY, Kang YH, Watanabe K, Tsai YC. 2012. The diversity of lactic acid bacteria in a traditional Taiwanese millet alcoholic beverage during fermentation. LWT-Food Sci Technol 51: 135-142.
Dave JM, Desai MV, Upadhayay KG, Thakar PN. 1980. Incidence of Enterococci in buffalo milk Cheddar cheese. GAU Research Journal 5 : 44-47.
Delavenne EJ, Mounier F, Déniel G. Barbier, G. Le Blay. 2012. Biodiversity of antifungal lactic acid bacteria isolated from raw milk samples from cow, ewe and goat over one-year period. Int J Food Microbiol 155: 185–190.
Elmoslih A, Zanzan M, Aissa R. 2017. Isolation and characterization of bacteriocinogenic Enterococcal and Lactococcal strains from south of Morocco dairy product. Biotechnol J Int 18(4): 1–16.
Franz CMAP, Van Belkum MJ, Holzapfel WH, Abriouel H, Gálvez A. 2007. Diversity of Enterococcal bacteriocins and their grouping in a new classification scheme. FEMS Microbiol Rev 31(3): 293–310.
Fusco V, Quero GM, Poltronieri P, Morea M, Baruzzi F. 2019. Autochthonous and probiotic lactic acid bacteria employed for production of “advanced traditional cheeses”. Foods 8 (9) : 412.
Gaaloul N, Ben Braiek O, Berjeaud JM. 2014. Evaluation of antimicrobial activity and safety aspect of Enterococcus italicus GGN10 strain isolated from Tunisian bovine raw milk. J Food Safety 34 (4): 300–311.
Gelsomino R, Vancanneyt M, Cogan TM, Condon S, Swings J. 2002. Source of Enterococci in a farmhouse raw-milk cheese. Appl Environ Microbiol 68(7): 3560–3565.
Georgieva R, Yocheva L, Tserovska L, Zhelezova G, Stefanova N, Atanasova A, Danguleva A, Ivanova G, Karapetkov N, Rumyan N, Karaivanova E. 2015. Antimicrobial activity and antibiotic susceptibility of Lactobacillus and Bifidobacterium spp. intended for use as starter and probiotic cultures. Biotechnol Biotechnol Equip 29(1): 84–91.
Gevers D, Huys G, Swings J. 2001. Applicability of repPCR fingerprinting for identification of Lactobacillus species. FEMS Microbiol Lett 205: 31-36.
Giménez-Pereira ML. 2005. Enterococci in Milk Products. [Dissertation]. Massey University Palmerston North, New Zealand. [New Zealand].
Goodman LB, Lawton MR, Franklin-Guild RJ, Anderson RR, Schaan L, Thachil AJ. 2017. Lactococcus petauri sp. nov., isolated from an abscess of a sugar glider. Int J Syst Evol Microbiol 67 (11) : 4397–4404.
Hakim NS, Suada IK, Sampurna IP. 2013. Ketahanan susu kuda Sumbawa pada penyimpanan suhu ruang ditinjau dari total asam, uji didih dan warna. IMV 2(4): 369-374.
Hanchi H, Mottawae W, Sebei K, Hammami R. 2018. The genus Enterococcus : between probiotic potential and safety concerns-an update. Front Microbiol 9: 1791.
Hermawati D. 2005. Kajian Aktivitas dan Karakterisasi Senyawa Antimikroba dari Susu Kuda Sumbawa. [Dissertation]. Pascasarjana Institut Pertanian Bogor, Bogor. [Indonesia].
Manguntungi B, Perkasa AS, Yulianti, Kusdianawati, Hastuti HP. 2018. Isolasi bakteri asam laktat dari susu kuda liar Sumbawa dan potensi antibakteri pada susu kuda liar Sumbawa. Biota 3 (2): 62-69.
Mikelsaar M, Zilmer M. 2009. Lactobacillus fermentum ME-3 an antimicrobial and antioxidative probiotic. Microb Ecol Health Dis 21(1): 1–27.
Mustopa AZ, Fatimah. 2014. Diversity of lactic acid bacteria isolated from Indonesian traditional fermented foods. Microbiology 8(2): 48-57.
Mustopa AZ. 2013. Isolation and Characterization of Lactobacillus plantarum S34 from Indonesia Traditional Food. [Dissertation]. danKOOK University, Korea. [Korea].
Morandi S, Silvetti T, Brasca M. 2013. Biotechnological and safety characterization of Enterococcus lactis, a recently described species of dairy origin. Anton Leeuw Int J G 103 (1) : 239–249.
Ou YJ, Ren QQ, Fang ST, Wu JG, Jiang YX, Chen YR, Zhong Y, Wang DD, Zhang GX. 2020. Complete genome insights into Lactococcus petauri CF11 isolated from a healthy human gut using second- and third-generation sequencing. Front Genet 11: 119.
Stiles ME, Holzapfel WH. 1997. Lactic acid bacteria of foods and their current taxonomy. Int J Food Microbiol 36 (1): 1-29.
Sujaya N, Ramona Y, Widarini NP, Suariani NP, Dwipayanti NMU, Nocianitri KA, Nursini NW. 2008. Isolasi dan karakterisasi bakteri asam laktat dari susu kuda Sumbawa. JVeteriner 9 (2) : 52-59.
Suneel D, Basappa K. 2013. Identification and characterization of Lactococcus garvieae and antimicrobial activity of its bacteriocin isolated from cow’s milk. Asian J Pharm Clin Res 6: 104-108.
Suzzi G, Caruso M, Gardini F, Lombardi A, Vannini L, Guerzoni ME, Andrighetto C, Lanorte MT. 2000. a survey of the enterococci isolated from an artisanal Italian goat's cheese (semicotto caprino). J Appl Microbiol 89 (2) : 267-74.
Tenea GN, Lara MI. 2019. Antimicrobial compounds produced by Weissella confusa Cys2-2 strain inhibit Gram-negative bacteria growth. CYTA-J Food 17(1): 105–111.
Villani F, Aponte M, Blaiotta G, Mauriello G, Pepe O, Moschetti G. 2001. Detection and characterization of a bacteriocin, garviecin L1-5, produced by Lactococcus garvieae isolated from raw cow’s milk. J Appl Microbiol 90 (3) : 430–439.
Welsh J, McClelland M. 1990. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18 (24) : 7213-8.
Widiada IGN, Antara NS, Aryanta WR. 2006. Identification and growth succession of lactic acid bacteria in wild horse milk of Bima during storage. In Annual Scientific Meeting of PERMI-2006. Udayana University, Bali. [Indonesia].
Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18 (22) : 6531-5.
Yu J, Wang WH, Menghe BL, Jiri MT, Wang HM, Liu WJ, Bao QH, Lu Q, Zhang JC, Wang F, Xu HY, Sun TS, Zhang HP. 2011. Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia. Int J Dairy Sci 94(7): 3229-41.
Zacharof MP, Lovitt RW. 2012. Bacteriocins produced by lactic acid bacteria. APCBEE Procedia 2: 50 - 56.
Zhu H, Qu F, Zhu LH. 1993. Isolation of genomic DNAs from plants, fungi, and bacteria using benzyl chloride. Nucleic Acids Res 21: 5279-5280.
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