Molecular Analysis of Lactic Acid Bacteria Isolated from Pado Fish of Agam Regency, West Sumatra Indonesia as Probiotics

نویسندگان
1 دانشکده علوم دامی، دانشگاه آندلاس
2 گروه فناوری پردازش محصولات حیوانی، دانشکده علوم دامی، دانشگاه آندلاس
3 بخش Farmasi، STIKES Prima اندونزی
4 گروه آگروتکنولوژی، دانشکده کشاورزی، دانشگاه آندلاس
5 گروه علوم و صنایع غذایی، دانشکده علوم، اجتماعی و آموزشی، دانشگاه پریما نوسانتارا بوکیتینگی
چکیده
Pado is a typical fermented fish from West Sumatra with a mixture of Simauang seed meat (Pangium edule Reinw) and coconut dregs covered with plastic for 4 to 8 days. Pado has good nutritional value and is thought to have a lactic acid bacterial activity to be used as a probiotic-producing functional food. Therefore, this study aims to determine the molecular isolation and identification of LAB types found in Pado fish using the 16S rRNA method. The research method is a descriptive method and laboratory analysis. The sample used as material for this research is Pado fish from Lubuk Basung District, Agam Regency, West Sumatera. The analysis related to probiotic characteristics includes total colonies of lactic acid bacteria, gram staining, acid resistance, resistance to bile salts, antimicrobial activity using test bacteria Escherichia coli O157, Staphylococcus aureus, Salmonella sp, and Listeria monocytogenes, as well as molecular sequential analysis using the 16S rRNA gene and phylogenetic diversity analysis using MEGA software version 7.0. The results showed that the total lactic acid bacteria is 7.3x107 CFU/gr; the seven isolates will be identified as gram-positive. Viability to gastric pH resistance ranged from 7.63-61.31%, and viability to bile salt resistance ranged from 22.86-71.05%. Three isolates had antimicrobial activity against E. coli, S. aureus, Salmonella sp., and L. monocytogenes. Molecular analysis using the 16S rRNA gene showed that PDY1 isolate is similar to Pediococcus acidilactici.
کلیدواژه‌ها

موضوعات


عنوان مقاله English

Molecular Analysis of Lactic Acid Bacteria Isolated from Pado Fish of Agam Regency, West Sumatra Indonesia as Probiotics

نویسندگان English

Yudha Pratama 1
Sri Melia 2
Endang Purwati 3
Yuherman Yuherman 2
Jamsari Jamsari 4
Hidayatul Jannah 5
1 Faculty of Animal Science, Andalas University
2 Department of Animal Products Processing Technology, Faculty of Animal Science, Andalas University
3 Departement of Farmasi, STIKES Prima Indonesia
4 Departement of Agrotechnology, Faculty of Agriculture, Andalas Univeristy
5 Department of Food Science and Technology, Faculty of Science, Social and Education, University of Prima Nusantara Bukittinngi
چکیده English

Pado is a typical fermented fish from West Sumatra with a mixture of Simauang seed meat (Pangium edule Reinw) and coconut dregs covered with plastic for 4 to 8 days. Pado has good nutritional value and is thought to have a lactic acid bacterial activity to be used as a probiotic-producing functional food. Therefore, this study aims to determine the molecular isolation and identification of LAB types found in Pado fish using the 16S rRNA method. The research method is a descriptive method and laboratory analysis. The sample used as material for this research is Pado fish from Lubuk Basung District, Agam Regency, West Sumatera. The analysis related to probiotic characteristics includes total colonies of lactic acid bacteria, gram staining, acid resistance, resistance to bile salts, antimicrobial activity using test bacteria Escherichia coli O157, Staphylococcus aureus, Salmonella sp, and Listeria monocytogenes, as well as molecular sequential analysis using the 16S rRNA gene and phylogenetic diversity analysis using MEGA software version 7.0. The results showed that the total lactic acid bacteria is 7.3x107 CFU/gr; the seven isolates will be identified as gram-positive. Viability to gastric pH resistance ranged from 7.63-61.31%, and viability to bile salt resistance ranged from 22.86-71.05%. Three isolates had antimicrobial activity against E. coli, S. aureus, Salmonella sp., and L. monocytogenes. Molecular analysis using the 16S rRNA gene showed that PDY1 isolate is similar to Pediococcus acidilactici.

کلیدواژه‌ها English

lactic acid bacteria
gene 16S rRNA
Pado
Probiotic
Pediococcus acidilacti
[1] Anisah, Harun, H., Jannah, H., Amelia, R., & Purwati, E. 2023. Molecular Identification and Antimicrobial Potency of Probiotic Lactic Acid Bacteria Pado (Fish Fermentation) Nagari Balingka IV Koto District-West Sumatra as a Functional Food. IOP Conference Series: Earth and Environmental Science, 1188(1). https://doi.org/10.1088/1755-1315/1188/1/012039
[2] Jannah, H., Syahminan, A. T. P., & Fajri, F. 2022. Sifat Kimia Pado, Ikan Fermentasi Tradisional Dari Kabupaten Agam, Sumatera Barat. Jurnal Peternakan~Borneo, 1(1), 1–5. https://doi.org/10.34128/jpb.v1i1.3
[3] Hasbullah, Kasim, A., Novelina, & Nurdin, H. 2016. Characterization traditional food Pado from west Sumatra. Der Pharmacia Lettre, 8(15), 26–31.
[4] Terpou, A., Papadaki, A., Lappa, I. K., Kachrimanidou, V., Bosnea, L. A., & Kopsahelis, N. 2019. Probiotics in Food Systems: Significance and Emerging Strategies Towards Improved Viability and Delivery of Enhanced Beneficial Value. Nutrients, 11(7), 1591. https://doi.org/10.3390/nu11071591
[5] Kristensen, M., Kalkman, G., Prevaes, S. M. P. J., Tramper-Stranders, G. A., Groot, K. M. de W., Janssens, H. M., Tiddens, H. A. W. M., Westreenen, M. van, Ent, C. K. van der, Sanders, E. A. M., Arets, B., Keijser, B., & Bogaert, D. 2016. WS07.5 Gut microbiome in healthy children and children with cystic fibrosis during the first 18 months of life. Journal of Cystic Fibrosis, 15, S12. https://doi.org/10.1016/s1569-1993(16)30100-x
[6] Melia, S., Purwati, E., Kurnia, YF & Pratama, DR. 2019. Antimicrobial potential of Pediococcus acidilactici from Bekasam, fermentation of sepat rawa fish (Tricopodus trichopterus) from Banyuasin, South Sumatra, Indonesia. J. Biodiversity. 12 (2): 3532-3538. DOI: 10.13057 / biodiv / d201210
[7] Pratama, Y. E., Jamsari, Yuherman, Melia, S., & Purwati, E. 2021. Molecular identification of lactic acid bacteria from "budu" of west Sumatra, Indonesia, as a potential probiotic. AACL Bioflux, Vol. 14 No (5), pg. 2837–2844.
[8] Devi D., Irawati N., Yanwirasti, Putra I. B., Pratama Y. E., 2023 Antimicrobial activities of lactic acid bacteriaLactobacillus casei isolated from “bekasam” fermented fish from “siak”. AACL Bioflux. Vol1 6 (6): pg.3193-3201.
[9] International Scientific Association for Probiotics and Prebiotics. 2009. Clarification of the Definition of a Probiotic. https://isappscience.org/about/mission-and-values/isapp-activities/#toggle-id-27. It was accessed on July 25, 2020.
[10] Food and Agriculture Organization / World Health Organization. 2002. Joint FAO / WHO Working Group Report on Drafting Guidelines for the Evaluation of Probiotics in Food, London.
[11] Purwati, E., Syukur, S & Hidayat, Z 2005. Lactobacillus sp. isolation from Biovicophitomega as a probiotic. Indonesian Institute of Sciences, Jakarta.
[12] Dwidjoseputro.1989. Microbiology basics. Djambat. Surabaya.
[13] Sunaryanto, R & Marwoto, B. 2013. Isolation, identification and characterization of lactic acid bacteria from buffalo milk curd. Indonesian Journal of Science and Technology, 14 (3): 228-233. DOI: https://doi.org/10.29122/jsti.v14i3.931.
[14] Yang, E., Fan, L., Jiang, Y., Doucette, C., & Fillmore, S. 2012. Antimicrobial activity of bacteriocin-producing lactic acid bacteria isolated from cheeses and yogurts. AMB Express, 2(1), 48. https://doi.org/10.1186/2191-0855-2-48.
[15] Irianto, K. 2008. Mikrobiologi: Menguak Dunia Mikroorganisme Jilid 2. Yrama Widya. Bandung.
[16] Salminen, S., Wright, A. V., & Ouwehand, A. 2004. Lactic acid bacteria: microbiological and functional aspect, 3rd Edition, Revised and Expanded. Marcel Dekker Inc, New York.
[17] Cotter, P. D., & Hill, C. 2003. Surviving the acid test: responses of gram-positive bacteria to low pH. Microbiology and molecular biology reviews : MMBR, 67(3), 429–453. https://doi.org/10.1128/MMBR.67.3.429-453.2003
[18] Abdullah, D., Poddar, S., Rai, R. P., Purwati, E., Dewi, N. P., & Pratama, Y. E. 2021. Molecular identification of lactic acid bacteria an approach to sustainable food security. Journal of public health research, 10(s2), jphr.2021.2508. https://doi.org/10.4081/jphr.2021.2508
[19] Roza, E., Aritonang, S.N., Yellita, Y., Susanty, H., Rizqan, Pratama, Y.E. 2021. Potential of dadiah kapau from Agam District, Indonesia as a source of probiotics for health. Biodiversitas 23: 564-571. https://doi.org/10.13057/biodiv/d230161
[20] Amelia, R., Phili,p K., Pratama, Y.E., Purwati, E. 2021.Characterization and probiotic potential of lactic acid bacteria isolated from dadiah sampledin West Sumatra. Food Sci Technol 41:746-752. DOI: 10.1590/fst.30020.
[21] Melia, S., Yuherman, Jaswandi, Purwati, E., Aritonang, S.N, and Silaen, M. 2017 Characterization of the Antimicrobial Activity of Lactic Acid Bacteria Isolated from Buffalo Milk in West Sumatra (Indonesia) Against Listeria monocytogenes. Mr. J Nutr 16 (8): 645-650. DOI: 10,393 / pjn. 2017,645,650.
[22] Pelczar, M. J., & Chan, E.C.S 2008. Basics of microbiology Volume I. UI Press, Jakarta.
[23] Syukur, S., Purwati, E., Urnemi & Jamsari. 2011. Screening and in-vitro antimicrobial protease activities from lactic acid bacteria associated with green cacao fermentation in West Sumatra, Indonesia. Proceedings of the International Seminar on IPR. Pekanbaru. 17-21.
[24] Hagstrom, A., Pinhassi, J & Zweifel, U. L. 2000. Biogeographical diversity among marine bacterioplankton. Aquat. Microb. E Col, 21: 231-244.
[25] Syafitri Y, Kusumaningrum H.D, & Hariyadi R.D. 2022. Identification of microflora and lactic acid bacteria in pado, a fermented fish productprepared with dried Pangium edule seed and grated coconut. Food Sci. Tech (Brazil). Vol 2. https://doi.org/10.1590/fst.19921