Molecular Identification of Lactic Acid Bacteria Approach to Sustainable food Security


Dessy Abdullah
Research Scholar, Lincoln University College, Wisma Lincoln, No, 12-18, SS 6/12, Off Jalan Perbandaran,, 47301 Petaling Jaya, Selangor D. E, Malaysia Lecturer, Medical Faculty, Baiturrahmah University Padang, West Sumatera, Indonesia, Indonesia.

Sandeep Poddar
Deputy Vice Chancellor of Research, Lincoln University College, Malaysia.
https://orcid.org/0000-0001-9771-877X

Ramesh Prasath Rai
Lincoln University College, Wisma Lincoln, No, 12-18, SS 6/12, Off Jalan Perbandaran, 47301 Petaling Jaya, Selangor, Malaysia, Malaysia.

Endang Purwati
Faculty of Animal Science, University of Andalas, Padang, West Sumatera, Indonesia, Indonesia.

Nadia Purnama Dewi
Research Scholar, Lincoln University College, Wisma Lincoln, No, 12-18, SS 6/12, Off Jalan Perbandaran,, 47301 Petaling Jaya, Selangor, Malaysia Lecturer, Medical Faculty, Baiturrahmah University Padang, West Sumatera, Indonesia, Indonesia.

Yudha Endra Pratama
Doctoral Program, Faculty of Animal Science, University of Andalas, Padang, Indonesia, Indonesia.

ABSTRACT

Background: Dadiah is a traditional dish from West Sumatra made from buffalo milk, which is fermented in bamboo tubes and left at room temperature for ±2 days. Dadiah is included in the staple food category because it contains Lactic Acid Bacteria (LAB) which has the potential to be a probiotic. This study aims to determine the identification and characterization of LAB from Dadiah from Halaban, Kab. Fifty Cities, West Sumatra.

Design and Methods: A survey method was used in this research with a descriptive analysis, Antimicrobial activity testing was done with bacteria Escherichia coli O157, Staphylococcus aureusListeria monocytogenes, and Listeria innocua. Molecular identification was done using the 16S rRNA gene.

Results: Probiotic candidate test with the best results in testing for resistance to stomach acid at pH3 with the viability of 65.98%, bile salt resistance 0.3%, viability of 54.90% from 2DA isolates. Antimicrobial activity with the best clear zone area results was obtained in 2DA isolates with Escherichia coli O157 test bacteria of 21.16 mm, Staphylococcus aureus with a clear zone area of 23.17 mm, Listeria innocua of 19.24 mm and Listeria monocytogenes with a clear zone area 18.23 mm in 4DA isolate, LAB identification using 16S sRNA gene, results of running PCR base length 1419bp.

Conclusions: Phylogenetic analysis shows that Dadiah of Limapuluh Kota Regency is a kin to Lactobacillus plantarum. The superiority of identification technology by using 16S rRNA gene only can be conducted if the nucleotide sequence information of the targeted bacteria is known beforehand.

 

REFERENCES

Tamang JP, editor. Ethnic fermented foods and alcoholic beverages of Asia. Delhi, India: Springer; 2016 Aug 5. DOI: https://doi.org/10.1007/978-81-322-2800-4

Surono IS. Indonesian dadih. Fermented milk and dairy products. 2015:377-99. Surono IS. Indonesian dadih. Fermented milk and dairy products. 2015:377-99.

Wirawati CU, Sudarwanto MB, Lukman DW, Wientarsih I. [Characteristic and Development of Cow’s Milk Dadih as an Alternate of Buffalo’s Milk Dadih. (Lukman DW, Wientarsih I. Karakteristik dan Pengembangan Dadih dari Susu Sapi sebagai Alternatif Dadih Susu Kerbau.)] [Article in Indonesian] Wartazoa 2017;27:95-103. DOI: https://doi.org/10.14334/wartazoa.v27i2.1595

WHO. Guidelines for the Evaluation of Probiotics in Food. Joint FAO/WHO Working Group Report on Drafting Guidelines for the Evaluation of Probiotics in Food London, Ontario, Canada, April 30 and May 1, 2002

Pratama DR, Pratama YE, Aritonang SN, et al. [Implementasi teknologi yogurt dengan menggunakan kultur bakteri isolat dadih asal nagari tanjung bonai untuk pemberdayaan peternak di jorong kayu maranting nagari tanjung bonai. (Implementation of yogurt technology using bacterial culture of curd isolate from Tanjung Bonai Village for the empowerment of farmers in Jorong Kayu Maranting Nagari Tanjung Bonai.)] [Article in Indonesian] The Downstream Science and Technology Journal. 2019 31;2:481-9.

Randazzo CL, Caggia C, Neviani E. Application of molecular approaches to study lactic acid bacteria in artisanal cheeses. Journal of Microbiological Methods. 2009;78:1-9. DOI: https://doi.org/10.1016/j.mimet.2009.04.001

Iranmanesh M, Ezzatpanah H, Mojgani N. Antibacterial activity and cholesterol assimilation of lactic acid bacteria isolated from traditional Iranian dairy products. LWT-Food Science and Technology 2014;58:355-9. DOI: https://doi.org/10.1016/j.lwt.2013.10.005

Kia KW, Arief II, Sumantri C, Budiman C. Plantaricin IIA-1A5 from Lactobacillus plantarum IIA-1A5 retards pathogenic bacteria in beef meatball stored at room temperature. Am J Food Technol 2016;11:37-43. DOI: https://doi.org/10.3923/ajft.2016.37.43

Juliyarsi I, Purwati E, Hartini P, et al. Characterization of lactic acid bacteria and determination of antimicrobial activity in tempoyak from Padang Pariaman District, West Sumatra, Indonesia. Pak J Nutr 2018;17:506. DOI: https://doi.org/10.3923/pjn.2018.506.511

Purwati E, Kurnia YF, Pratama DR. Antimicrobial potential of Pediococcus acidilactici from Bekasam, fermentation of sepat rawa fish (Tricopodus trichopterus) from Banyuasin, South Sumatra, Indonesia. Biodiversitas 2019;20:1210 DOI: https://doi.org/10.13057/biodiv/d201210

Melia S, Purwati E, Yuherman J, et al. Characterization of the antimicrobial activity of lactic acid bacteria isolated from buffalo milk in West Sumatera (Indonesia) against Listeria monocytogenes. Pak J Nutr 2017;16:645-50. DOI: https://doi.org/10.3923/pjn.2017.645.650

Harun H, Wirasti Y, Purwanto B, Purwati E. Characterization of lactic acid bacteria and determination of antimicrobial activity in dadih from Air Dingin Alahan Panjang District, Solok Regency-West Sumatera. Systematic Rev Pharm 2020;11.

Tsai CC, Lin PP, Hsieh YM, et al. Cholesterol-lowering potentials of lactic acid bacteria based on bile-salt hydrolase activity and effect of potent strains on cholesterol metabolism in vitro and in vivo. Sci World J 2014;2014:690752. DOI: https://doi.org/10.1155/2014/690752

Liévin-Le Moal V, Servin AL. Anti-infective activities of lactobacillus strains in the human intestinal microbiota: from probiotics to gastrointestinal anti-infectious biotherapeutic agents. Clinical Microbiol Rev 2014;27:167-99. DOI: https://doi.org/10.1128/CMR.00080-13

Lee BH, Lo YH, Pan TM. Anti-obesity activity of Lactobacillus fermented soy milk products. J Funct Foods 2013;5:905-13. DOI: https://doi.org/10.1016/j.jff.2013.01.040

Kuda T, Kawahara M, Nemoto M, et al. In vitro antioxidant and anti-inflammation properties of lactic acid bacteria isolated from fish intestines and fermented fish from the Sanriku Satoumi region in Japan. Food Res Int 2014;64:248-55. DOI: https://doi.org/10.1016/j.foodres.2014.06.028

Kawabata K, Kato Y, Sakano T, et al. Effects of phytochemicals on in vitro anti-inflammatory activity of Bifidobacterium adolescentis. Biosci Biotechnol Biochem 2015;79:799-807. DOI: https://doi.org/10.1080/09168451.2015.1006566

Ahmadi MA, Ebrahimi MT, Mehrabian S, et al. Antimutagenic and anticancer effects of lactic acid bacteria isolated from Tarhana through Ames test and phylogenetic analysis by 16S rDNA. Nutrition Cancer 2014;66:1406-13. DOI: https://doi.org/10.1080/01635581.2014.956254

Apás AL, González SN, Arena ME. Potential of goat probiotic to bind mutagens. Anaerobe 2014;28:8-12. DOI: https://doi.org/10.1016/j.anaerobe.2014.04.004

Haghshenas B, Nami Y, Abdullah N, et al. Anticancer impacts of potentially probiotic acetic acid bacteria isolated from traditional dairy microbiota. LWT-food Sci Technol 2015;60:690-7. DOI: https://doi.org/10.1016/j.lwt.2014.09.058

Fasseas MK, Fasseas C, Mountzouris KC, Syntichaki P. Effects of Lactobacillus salivarius, Lactobacillus reuteri, and Pediococcus acidilactici on the nematode Caenorhabditis elegans include possible antitumor activity. Appl Microbiol Biotechnol 2013;97:2109-18. DOI: https://doi.org/10.1007/s00253-012-4357-9

Tillisch K, Labus J, Kilpatrick L, et al. Consumption of fermented milk product with probiotic modulates brain activity. Gastroenterol 2013;144:1394-401. DOI: https://doi.org/10.1053/j.gastro.2013.02.043

Legarová V, Kouřimská L. [Sensory quality evaluation of whey-based beverages. (Mljekarstvo: časopis za unaprjeđenje proizvodnje i prerade mlijeka)]. Mljekarstvo 2010;60:280-7.

Pescuma M, Hébert EM, Mozzi F, De Valdez GF. Functional fermented whey-based beverage using lactic acid bacteria. Int J Food Microbiol 2010;141:73-81. DOI: https://doi.org/10.1016/j.ijfoodmicro.2010.04.011

Ashraf R, Vasiljevic T, Day SL, et al. Lactic acid bacteria and probiotic organisms induce different cytokine profile and regulatory T cells mechanisms. J Functional Foods 2014;6:395-409. DOI: https://doi.org/10.1016/j.jff.2013.11.006

Cox CM, Dalloul RA. Immunomodulatory role of probiotics in poultry and potential in ovo application. Beneficial Microbes 2015;6:45-52. DOI: https://doi.org/10.3920/BM2014.0062

Ren D, Li C, Qin Y, et al. In vitro evaluation of the probiotic and functional potential of Lactobacillus strains isolated from fermented food and human intestine. Anaerobe 2014;30:1-0. DOI: https://doi.org/10.1016/j.anaerobe.2014.07.004

Lee DK, Jang S, Baek EH, et al. Lactic acid bacteria affect serum cholesterol levels, harmful fecal enzyme activity, and fecal water content. Lipids Health Disease 2009;8:21. DOI: https://doi.org/10.1186/1476-511X-8-21

Sharma R, Kapila R, Kapasiya M, et al. Dietary supplementation of milk fermented with probiotic Lactobacillus fermentum enhances systemic immune response and antioxidant capacity in aging mice. Nutrition Res 2014;34:968-81. DOI: https://doi.org/10.1016/j.nutres.2014.09.006

Kwak SH, Cho YM, Noh GM, Om AS. Cancer preventive potential of kimchi lactic acid bacteria (Weissella cibaria, Lactobacillus plantarum). J Cancer Prev 2014;19:253. DOI: https://doi.org/10.15430/JCP.2014.19.4.253

Fung WY, Liong MT. Evaluation of proteolytic and ACE-inhibitory activity of Lactobacillus acidophilus in soy whey growth medium via response surface methodology. LWT-Food Sci Technol 2010;43:563-7. DOI: https://doi.org/10.1016/j.lwt.2009.10.004

Rodríguez-Figueroa JC, González-Córdova AF, Astiazaran-García H, et al. Antihypertensive and hypolipidemic effect of milk fermented by specific Lactococcus lactis strains. J Dairy Sci 2013;96:4094-9. DOI: https://doi.org/10.3168/jds.2012-6014

Chen Y, Liu W, Xue J, et al. Angiotensin-converting enzyme inhibitory activity of Lactobacillus helveticus strains from traditional fermented dairy foods and antihypertensive effect of fermented milk of strain H9. J Dairy Sci 2014;97:6680-92. DOI: https://doi.org/10.3168/jds.2014-7962

Public Health England. Identification of Staphylococcus Species, Micrococcus Species and Rothia Species. Standards Unit, Microbiology Services, PHE. 2014.

Rashid S, Hassanshahian M. Screening, isolation and identification of lactic acid bacteria from a traditional dairy product of Sabzevar, Iran. Int J Enteric Pathog 2014;2:1-5. DOI: https://doi.org/10.17795/ijep18393

Yu J, Wang WH, Menghe BL, et al. Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia. J Dairy Sci 2011;94:3229-41. DOI: https://doi.org/10.3168/jds.2010-3727

Yang Z, Rannala B. Molecular phylogenetics: principles and practice. Nature Rev Gen 2012;13:303-14. DOI: https://doi.org/10.1038/nrg3186

FAO/WHO. Report on Joint FAO/WHO Expert Consultation on Evaluation of Health and Nutritional Properties of Probiotics in Food Including Powder Milk with Live Lactic Acid Bacteria. FAO/WHO, 2001.

Amelia. R, Philip K, Pratama YE, Purwati E. Characterization and probiotic potential of lactic acid bacteria isolated from dadiah sampled in West Sumatra. Food Sci Technol 2020 Dec. https://doi.org/10.1590/fst.30020 DOI: https://doi.org/10.1590/fst.30020

Djide MN, Wahyudin E. [Isolasi Bakteri Asam Laktat dari Air Susu Ibu, dan Potensinya dalam penurunan kadar kolesterol secara in vitro. (Isolation of lactic acid bacteria from mother's milk, and its potential in lowering cholesterol levels in vitro.)] [Article in Indonesian] Pharmacy and Pharmacology Magazine 2008;12:73-8.

Nurnaafi A, Setyaningsih I. [Potensi Probiotik Bakteri Asam Laktat Asal Bekasam Ikan Nila. (Probiotic Potential of lactic acid bacteria from tilapia. )] Journal of Food Technology and Industry 2015;26:109-14. DOI: https://doi.org/10.6066/jtip.2015.26.1.109

Schoustra SE, Kasase C, Toarta C, et al. Microbial community structure of three traditional zambian fermented products: mabisi, chibwantu and munkoyo. PloS One 2013;8:e63948. DOI: https://doi.org/10.1371/journal.pone.0063948

Purwati E, Syukur S, Hidayat Z. [Lactobacillus sp. Isolasi dari biovicophitomega sebagai probiotik. (Lactobacillus sp. Isolation of biovicophitomega as probiotic.)] Proceedings of the Indonesian Institute of Sciences, Jakarta, 2005:24-5.

Suryani Y, Astuti OB, Umniyati S. [Isolasi dan karakterisasi bakteri asam laktat dari limbah kotoran ayam sebagai agensi probiotik dan enzim kolesterol reduktase. (Isolation and characterization of lactic acid bacteria from chicken manure as probiotic agents and cholesterol reductase enzymes.)] [Article in Indonesian] Proceedings of the National Biology Seminar. 2010;138-147.

Salminen S, Von Wright A, eds. Lactic acid bacteria: microbiological and functional aspects. CRC Press; 2004 Jul 23. DOI: https://doi.org/10.1201/9780824752033.ch7

Unus U. Basic Microbiology. Papas Sinar Sinanti, Jakarta. 2005.

Sunaryanto R, Marwoto B. [Isolasi, identifikasi dan karakterisasi bakteri asam laktat dari dadih susu kerbau. (Isolation, identification, and characterization of lactic acid bacteria from buffalo milk curd.)] [Article in Indonesian] Indonesian Journal of Science and Technology. 2013;14:228-33. DOI: https://doi.org/10.29122/jsti.v14i3.931

Purwati E, Syukur S, Husmaini, Purwanto H, Pasaribu R P. [Molekuler Karakterisasi Bakteri Asam Laktat Isolate Dadih Air Dingin Kabupaten Solok Sumatera Barat. (Molecular characterization of lactic acid bacteria isolate from cold water curd Solok Regency, West Sumatra.)] [Article in Indonesian] Journal of Innovation Research 2014;40:l3l-46

Nur F, Hafsan H, Wahdiniar A. [Isolasi bakteri asam laktat berpotensi probiotik pada dangke, makanan tradisional dari susu kerbau di Curio Kabupaten Enrekang. (Isolation of lactic acid bacteria with probiotic potential in dangke, traditional food from buffalo milk in Curio, Enrekang Regency.)] [Article in Indonesian] Biogenesis: Scientific Journal of Biology. 2015;3:60-5. DOI: https://doi.org/10.24252/bio.v3i1.568

Bharwade M, Balakrishnan S, Chaudhary N, Jain AK. Fatty Acid Profile and Physico-Chemical Characteristics of Milk Lipids of Kankrej Cow. Int J Current Microbiol Appl Sci 2017;6:3035-47. DOI: https://doi.org/10.20546/ijcmas.2017.608.363

Melia S, Ferawati Y, Jaswandi PH, Purwati E. Probiotic characterization of lactic acid bacteria isolated from raw milk (buffalo, cow, and goat) from West Sumatera, Indonesia. Asian J Microbiol Biotechnol Environ Sci 2018;20:S131-9.

Shafakatullah N, Chandra M. Screening of raw buffalo’s milk from Karnataka for potential probiotic strains. Res J Rec Sci 2014;3:2502.

Anandharaj M, Sivasankari B. Isolation of potential probiotic Lactobacillus oris HMI68 from mother's milk with cholesterol-reducing property. J Biosci Bioeng 2014;118:153-9. DOI: https://doi.org/10.1016/j.jbiosc.2014.01.015

Arnold M, Rajagukguk YV, Gramza-Michałowska A. Characterization of Dadih: Traditional Fermented Buffalo Milk of Minangkabau. Beverages 2021;7:60. DOI: https://doi.org/10.3390/beverages7030060