Molecular Isolation, Probiotic Property, and Bacteriocin Production of Enterococcus faecium (TM81) and Lactobacillus curvatus (TM51) with Anti-Listerial Activity in Native Dairy Products of Iran

Document Type : Research Article

Authors

1 Department of Food Hygiene and Aquaculture, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN

2 Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN

Abstract

Lactic acid bacteria in dairy products are the important bacteriocin-producing group with an antibacterial activity effect. The aim of this study was the isolation and molecular identification of bacteriocin-producing enterococci and lactobacilli with a wide range of antibacterial activity. Bacteriocin-producing bacteria were isolated and purified from Iranian native dairy products. The effect of bacteriocins on the Gram-negative and Gram-positive indicator strains, as well as the impact of proteolytic enzymes, pH, and temperature changes on bacteriocin production, were investigated. A total of 431 bacteriocin-producing isolates were characterized from 100 traditional cheese and yoghurt samples. The resulting bacteriocin reduced the growth of these two strains. E. faecium TM81 had both bactericidal and bacteriostatic properties, whereas the L. curvatus TM51 only had bacteriostatic properties. The bacteriocin produced by both strains had the highest antibacterial effect among Gram-positive indicator strains such as Listeria monocytogenes and Bacillus cereus. Because produced bacteriocin (resistant to temperature and pH changes) has a wide range of inhibitory spectra on Gram-positive and Gram-negative bacteria, especially pathogenic bacteria, it is recommended to use it as a bio preservative in the food/pharmaceutical industry, and animal feed, as well as an alternative to chemical antibiotics.

Keywords

Main Subjects


[1] Goodarzi A., Hovhannisyan H., Barseghyan A., Elimination of Pathogen Escherichia coli O157:H7
in Ground Beef by a Newly Isolated Strain of Lactobacillus acidophilus during Storage at 5°C
. Appl. Food Biotechnol., 3(3): 170-176 (2016).
[4] Zoghi A., Khosravi-Darani K., Sohrabvandi S., Attar H., Alavi S., Survival of Probiotics in Synbiotic Apple Juice During Refrigeration and Subsequent Exposure to Simulated Gastro-Intestinal Conditions. Iran. J. Chem. Chem. Eng. (IJCCE), 38(2): 159-170 (2019).
 [6] Hasan Hussein F., Razavi S., Emam Djomeh Z. Evaluation of Physicochemical, Sensorial and Microbiological Attributes of Fermented Camel Sausages. Iran. J. Chem. Chem. Eng. (IJCCE), 38(2): 171-181 (2019).
[7] Tagg, J.R., Dajani, A.S., Wannamaker, L.W., Bacteriocins of Gram-positive bacteria. Bacteriol. Rev., 40(3): 722 (1976).
[8] De Vuyst L., Leroy F., Bacteriocins from Lactic Acid Bacteria: Production, Purification, and Food Applications. J. Mol. Microbiol. Biotechnol., 13(4): 194-199 (2007).
[9] Jandaghi H., Ghahramani Seno M., Farzin H., Mohsenzadeh M., Rapid Quantitative Detection of Listeria monocytogenes in Chicken Using Direct and Combined Enrichment/qPCR Method. Iran. J. Chem. Chem. Eng. (IJCCE), Article in Press, (2019).
[10] Weisburg W.G., Barns S.M., Pelletier D.A., Lane D.J., 16S Ribosomal DNA Amplification for Phylogenetic Study. J. Bacteriol., 173(2): 697-703 (1991).
[11] Karthikeyan V., Santhosh S.W., Study of Bacteriocin as a Food Preservative and the L. acidophilus Strain as Probiotic. Pak. J. Nutr., 8: 335-340 (2009).
[12]  Schillinger, U., Lucke, F.K., Antibacterial Activity of Lactobacillus sake Isolated from Meat. Appl. Environ. Microbiol., 55(8): 1901-1906 (1989).
[13] Yamamoto, Y., Togawa, Y., Shimosaka, M., Okazaki, M., Purification and Characterization of Novel Bacteriocin Produced by Enterococcus faecalis Strain RJ-11. Appl. Environ. Microbiol., 69(10): 5746-5753 (2003).
 [14] Magnusson, J., & Schnurer, J., Lactobacillus coryniformis Subsp. coryniformis Strain Si3 Produces a Broad-Spectrum Proteinaceous Antifungal Compound. Appl. Environ. Microbiol., 67(1): 1-5 (2001).
[15] Shokria D., Zaghian S., Fazeli H., Mobasherizadeh S., Ataei B., Isolation and Purification of an Ultraviolet-Stable Bacteriocin Produced by Enterococcus Faecium Strain DSH20 Against Listeria Monocytogenes. J. Isfahan Med. School., 31: 236 (2013).
[18] Todorov S.D., Wachsman M., Tomé E., Dousset X., Destro M.T., Dicks L.M.T., de Melo Franco, B.D.G., Vaz-Velho, M., Drider, D., Characterisation of an Antiviral Pediocin-Like Bacteriocin Produced by Enterococcus faecium. Food Microbiol., 27(7): 869-879 (2010).
[19] Sonsa-Ard N., Rodtong S., Chikindas M.L., Yongsawatdigul J., Characterization of Bacteriocin Produced by Enterococcus faecium CN-25 Isolated from Traditionally Thai Fermented Fish Roe. Food Control., 54: 308-316 (2015).
[20]  Saranya S., Hemashenpagam N., Antagonistic Activity and antibiotic sensitivity of lactic acid bacteria from Fermented Dairy Products. Adv. Appl. Sci. Res., 2(4): 528-534 (2011).
[21] Adetunji V.O., Adegoke G.O., Bacteriocin and Cellulose Production by Lactic Acid Bacteria Isolated from West African Soft Cheese. Afr. J. Biotechnol., 6(22): 2616-2619 (2007).
[22] CLSI. "Performance Standards for Antimicrobial Susceptibility Testing". 29th ed. CLSI supplement M100. Wayne, PA: Clinical and Laboratory Standards Institute; 2019.
[24] Aymerich T., Holo H., Havarstein L.S., Hugas M., Garriga M., Nes I.F., Biochemical and Genetic Characterization of enterocin A from Enterococcus faecium, a New Antilisterial Bacteriocin in the Pediocin Family of Bacteriocins. Appl. Environ. Microbiol., 62(5): 1676-1682 (1996).
[25] Toit M.D., Franz C.M.A.P., Dicks L.M.T., Holzapfel, W.H., Preliminary Characterization of Bacteriocins Produced by Enterococcus faecium and Enterococcus faecalis Isolated from Pig Faeces. J. Appl. Microbiol., 88(3): 482-494 (2000).
[26] Gutiérrez J., Criado R., Citti R., Martín  M., Herranz  C., Nes  I.F., Cintas  L.M., Hernández P.E., Cloning, Production and Functional Expression of enterocin P, a Sec-Dependent Bacteriocin Broduced by Enterococcus Faecium P13, in Escherichia coli. Int. J. Food Microbiol., 103(3): 239-250 (2005)
[27] Hadji-Sfaxi I., El-Ghaish S., Ahmadova A., Batdorj B., Le Blay-Laliberté G., Barbier G., Ha ertlé T., Chobert J.M., Antimicrobial Activity and Safety of Use of Enterococcus faecium PC4. 1 Isolated from Mongol Yogurt. Food Control., 22(12): 2020-2027 (2011).
[28] Moreno M.F., Sarantinopoulos P., Tsakalidou E., De Vuyst L., The Role and Application of Enterococci in Food and Health. Int.J. Food Microbiol., 106(1): 1-24 (2006).
[29] Todorov, S.D., Bacteriocins from  Lactobacillus plantarum  Production, Genetic Organization and Mode of Action. Produção, Organização Genética e Modo de Ação. A Review. Braz. J. Microbiol., 40(2): 209-221 (2009).
[30] Ahmadova A., Todorov S.D., Choiset Y., Rabesona H., Mirhadizadi T., Kuliyev A., de Melo Franco, B.D.G., Chobert, J.M., Haertlé, T., Evaluation of Antimicrobial Activity, Probiotic Properties and Safety of Wild Strain Enterococcus faecium AQ71 Isolated from Azerbaijani Motal Cheese. Food Control., 30(2): 631-641 (2013).
[31] Giraffa G., Functionality of enterococci in Dairy Products. Int. J. Food Microbiol., 88(2-3): 215-222 (2003).
[32] Veljovic K., Fira D., Terzic-Vidojevic A.,
Abriouel H., Galvez A., Topisirovic L., Evaluation of Antimicrobial and Proteolytic Activity of Enterococci Isolated from Fermented Products, Eur. Food Res. Technol., 230(1): 63-70 (2009).
[33] Barbosa J., Ferreira V., Teixeira P., Antibiotic Susceptibility of Enterococci Isolated from Traditional Fermented Meat Products, Food Microbiol., 26(5): 527-532 (2009).
[34] Ammor M.S., Flórez A.B., Mayo B., Antibiotic Resistance in Non-Enterococcal Lactic Acid Bacteria and Bifidobacteria. Food Microbiol., 24(6): 559-570 (2007).
[35] Nithya K., Senbagam D., Senthilkumar B., Udhayashree N., Gurusamy R., Characterization of Bacteriocin Producing Lactic Acid Bacteria and its Application as a Food Preservative. Afr. J. Microbiol. Res., 6(6): 1138-1146 (2012).
[36] Rajaram G., Manivasagan P., Thilagavathi B., Saravanakumar A., Purification and Characterization of a Bacteriocin Produced by Lactobacillus lactis Isolated from Marine Environment. Adv. J. Food Sci. Technol., 2(2): 138-144 (2010).
[37] Carminati D., Giraffa G., Quiberoni A., Binetti A., Suárez V., Reinheimer J., Advances and Trends in Starter Cultures for Dairy Fermentations. In: "Biotechnol. of Lactic Acid Bacteria: Novel Applications", John Wiley and Sons, p. 177-192 (2010).
[38] Hébert L., Courtin P., Torelli R., Sanguinetti M., Chapot-Chartier M.P., Auffray Y., Benachour, A., Enterococcus Faecalis Constitutes an Unusual Bacterial Model in Lysozyme Resistance. Infect. Immun. 75(11): 5390-5398 (2007).
[39] Aleksieva V., Enterococcal and Coliform Content in White Brine Cheese. Veterinarno-Meditsinski Nauki. 17(2): 85-91 (1980).
[40] Aymerich T., Holo H., Håvarstein L.S., Hugas M., Garriga M., Nes I.F., Biochemical and Genetic Characterization of Enterocin a from Enterococcus Faecium, a New Antilisterial Bacteriocin in the Pediocin Family of Bacteriocins, Appl. Environ. Microbiol., 62(5): 1676-1682 (1996).
[41] Özdemir G.B., Oryaşın E., Bıyık H.H., Özteber M., Bozdoğan B., Phenotypic and Genotypic Characterization of Bacteriocins in Enterococcal Isolates of Different Sources. Indian J. Microbiol., 51(2): 182-187 (2011).
[42] Edalatian M.R., Najafi M.B.H., Mortazavi S.A., Alegría Á., Delgado S., Bassami M.R., Mayo, B., Production of Bacteriocins by Enterococcus spp. Isolated fromTraditional, Iranian, Raw Milk Cheeses, and Detection of Their Encoding Genes. Eur. Food Res. Technol., 234(5): 789-796 (2012).
[43] Nascimento M.D.S.D., Moreno I., Kuaye A.Y., Antimicrobial Activity of Enterococcus faecium FAIR-E 198 Against Gram-Positive Pathogens. Braz. J. Microbiol., 41(1): 74-81 (2010).
[44] Chen Y.S., Yanagida F., Srionnual S., Characteristics of Bacteriocin‐Like Inhibitory Substances from Dochi‐Isolated Enterococcus faecium D081821 and D081833. Letters in Appl. Microbiol., 44(3): 320-325 (2007).
[45] Aran H., Biscola V., El-Ghaish S., Jaffres E., Dousset X., Pillot G., Haertlé T., Chobert J.M., Hwanhlem, N., Bacteriocin-producing Enterococcus faecalis KT2W2G Isolated from Mangrove Forests in Southern Thailand: Purification, Characterization and Safety Evaluation. Food Control., 54: 126-134 (2015).