The Study of the Probiotic Potential of the Beneficial Bacteria Isolated from Kefir Grains

Open access

Abstract

The aim of this study was to identify beneficial bacteria with probiotic potential from kefir grains. The lactobacilli isolated from kefir grains were characterised as: Lactobacillus plantarum, Lactobacillus paraplantarum, Lactobacillus paracasei, and Lactobacillus kefiri. The strains Lb. plantarum 1Ž, Lb. paraplantarum S10, and Lb. paracasei 2Ž tolerated better the test gastric juice at pH 2 and 2.6 during 120 min of incubation in comparison with the strains Lb. kefiri. On the other hand, the strains Lb. kefiri were resistant to 0.3 % bile acid salts. The Lb. paracasei 2Ž showed the significantly highest survival (P < 0.001) at pH 2 in comparison with all other strains tested and was also able to tolerate 0.3 % concentration of the bile salts. All strains produced medium to strong biofilms on abiotic surfaces and inhibited the growth of selected potential pathogens with varying intensity. All kefir isolates were susceptible to the antibiotics tested and exhibited positive β-galactosidase activity with the exception of Lb. paracasei 2Ž which did not show any activity of undesirable enzymes, such as β-glucosidase and β-glucuronidase. Additional testing and validation of the biological properties and safety of the strain Lb. paracasei 2Ž under in vivo conditions are needed to confirm the prospective use of this strain in practice.

1. Anas, M., Eddine, H. J., Mebrouk, K., 2008: Antimicrobial activity of Lactobacillus species isolated from Algerian raw goat’s milk against Staphylococcus aureus. World Journal of Dairy and Food Sciences, 3, 39—49.

2. Angulo, L., Lopez, E., Lema, C., 1993: Microflora present in kefir grains of the Galician region (North-West of Spain). J. Dairy Res., 60, 263—267.

3. Bessede, E., Angla-Gre, M., Delagarde, Y., Sep Hieng, S., Ménard, A., Mégraud, F., 2011: Matrix-assisted laser-desorption/ionization biotyper: experience in the routine of a university hospital. Clin. Microbiol. Infect., 17, 533—538.

4. Bilkova, A., Kinova Sepova, H., Bukovsky, M., Bezakova, L., 2011: Antibacterial potential of lactobacilli isolated from a lamb. Vet. Med. Czech, 56, 319—324.

5. Carasi, P., Díaz, M., Racedo, S. M., De Antoni, G., Urdaci, M. C., Serradell, Mde. L., 2014: Safety characterization and antimicrobial properties of kefir-isolated Lactobacillus kefiri. Biomed. Res. Int., 2014, Article ID 208974, 7. http://dx.doi.org/10.1155/2014/208974.

6. Chaieb, K., Chehab, O., Zmantar, T., Rouabhia, M., Mahdouani, K., Bakhrouf, A., 2007: In vitro effect of pH and ethanol on biofilm formation by clinical ica-positive Staphylococcus epidermidis strains. Ann. Microbiol., 57, 431—437.

7. Chen, Y. P., Hsiao, P. J., Hong, W. S., Dai, T. Y., Chen, M. J., 2012: Lactobacillus kefiranofaciens M1 isolated from milk kefir grains ameliorates experimental colitis in vitro and in vivo. J. Dairy Sci., 95, 63—74.

8. Cheow, W. S., Hadinoto, K., 2013: Biofilm-like Lactobacillus rhamnosus probiotics encapsulated in alginate and carrageenan microcapsules exhibiting enhanced thermotolerance and freeze-drying resistance. Biomacromolecules, 14, 3214—3222.

9. Chifiriuc, M. C., Cioaca, A. B., Lazar, V., 2011: In vitro assay of the antimicrobial activity of kephir against bacterial and fungal strains. Anaerobe, 17, 433—435.

10. Danielsen, M., Wind, A., 2003: Susceptibility of Lactobacillus spp. to antimicrobial agents. Int. J. Food Microbiol., 82, 1—11.

11. De Smet, I., Van Hoorde, L., Van De Woestyne, M., Christiaens, H., Verstraete, W., 1995: Significance of bile salt hydrolytic activities of lactobacilli. J. Appl. Bacteriol., 79, 292—301.

12. Delgado, S., O’Sullivan, E., Fitzgerald, G., Mayo, B., 2008: In vitro evaluation of the probiotic properties of human intestinal Bifidobacterium species and selection of new probiotic candidates. J. Appl. Microbiol., 104, 1119—1127.

13. Duwat, P., Cesselin, B., Sourice, S., Gruss, A., 2000: Lactococcus lactis, a bacterial model for stress responses and survival. Int. J. Food Microbiol., 55, 83—86.

14. EFSA, 2012: Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. The EFSA Journal, 10, 2740, 10 pp.

15. Elli, M., Zink, R., Rytz, A., Reniero, R., Morelli, L., 2000: Iron requirement of Lactobacillus spp. in completely chemically defined growth media. J. Appl. Microbiol., 88, 695—703.

16. Essid, I., Medin, M., Hassouna, M., 2009: Technological and safety properties of Lactobacillus plantarum strains isolated from a Tunisian traditional salted meat. Meat Sci., 81, 203—208.

17. FAO/WHO, 2006: Probiotic in foods. Health and nutritional properties and guidelines for evaluation. FAO Food Nutr., 85.

18. Fernández, M. F., Boris, S., Barbés, C., 2003: Probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract. J. Appl. Microbiol., 94, 449—455.

19. Garrote, G. L., Abraham, A. G., De Antoni, G. L., 2001: Chemical and microbiological characterization of kefir grains. J. Dairy Res., 68, 639—652.

20. Hertzler, S. R., Clancy, S. M., 2003: Kefir improves lactose digestion and tolerance in adults with lactose maldigestion. J. Am. Diet. Assoc., 103, 582—587.

21. Hong, W. S., Chen, H. C., Chen, Y. P., Chen, M. J., 2009: Effects of kefir supernatant and lactic acid bacteria isolated from kefir grain on cytokine production by macrophage. Int. Dairy J., 19, 244—251.

22. Husain, S., 2008: Effect of ferric iron on siderophore production and pyrene degradation by Pseudomonas fluorescens 29L. Curr. Microbiol., 57, 331—334.

23. Hütt, P., Shchepetova, J., Loivukene, K., Kullisaar, T., Mikelsaar, M., 2006: Antagonistic activity of probiotic lactobacilli and bifidobacteria against entero- and uropathogens. J. Appl. Microbiol., 100, 1324—1332.

24. Jones, S. E., Versalovic, J., 2009: Probiotic Lactobacillus reuteri biofilms produce antimicrobial and anti-inflammatory factors. BMC Microbiol., 9, 35—43.

25. Kõll, P., Mändar, R., Smidt, I., Hütt, P., Truusalu, K., Mikelsaar, R. H. et al., 2010: Screening and evaluation of human intestinal lactobacilli for the development of novel gastrointestinal probiotics. Curr. Microbiol., 61, 560—566.

26. Kos, B., Šušković, J., Goreta, J., Matošić, S., 2000: Effect of protectors on the viability of Lactobacillus acidophilus M92 in simulated gastrointestinal conditions. Food Technology and Biotechnology, 38, 121—127.

27. Kubota, H., Senda, S., Nomura, N., Tokuda, H., Uchiyama, H., 2008: Biofilm formation by lactic acid bacteria and resistance to environmental stress. Journal of Bioscience and Bioengineering, 106, 381—386.

28. Lee, Y. K., Salminen, S., 2009: Handbook of Probiotics and Prebiotics. 2nd edn., John Wiley & Sons, Inc., Hoboken, New Jersey, published simultaneously in Canada, 596 pp.

29. Leite, A. M. O., Miguel, M. A. L., Peixoto, R. S., Rosado, A. S., Silva, J. T., Paschoalin, V. M. F., 2013: Microbiological, technological and therapeutic properties of kefir: a natural probiotic beverage. Braz. J. Microbiol., 44, 341—349.

30. Macfarlane, S., Bahrami, B., Macfarlane, G. T., 2011: Mucosal biofilm communities in the human intestinal tract. Adv. Appl. Microbiol., 75, 111—143.

31. Maragkoudakis, P. A., Zoumpopoulou, G., Miaris, Ch., Kalantzopoulos, G., Pot, B., Tsakalidou, E., 2006: Probiotic potential of Lactobacillus strains isolated from dairy products. Int. Dairy J., 16, 189—199.

32. Martín, R., Soberón, N., Vaneechoutte, M., Flórez, A. B., Vázquez, F., Suárez, J. E., 2008: Characterization of indigenous vaginal lactobacilli from healthy women as probiotic candidates. Int. Microbiol., 11, 261—266.

33. Mathur, S., Singh, R., 2005: Antibiotic resistance in food lactic acid bacteria − a review. Int. J. Food Microbiol., 105, 281—295.

34. Mattila-Sandholm, T., 2002: Technological challenges for future probiotic foods. Int. Dairy J., 12, 173—182.

35. Nemcová, R., 2009: Lactic acid bacteria from the point of view of transfer of resistance to antibiotics (In Slovak). Slovenský veterinársky časopis (Slovak Veterinary Journal), 34, 16—20.

36. Niazi Amraii, H., Abtahi, H., Jafari, P., Mohajerani, H. R., Fakhroleslam, M. R., Akbari, N., 2014: In vitro study of potentially probiotic lactic acid bacteria strains isolated from traditional dairy products. Jundishapur Journal of Microbiology, 7, e10168.

37. Nielsen, B., Gurakan, G. C., Unlu, G., 2014: Kefir: A multifaceted fermented dairy product. Probiotics. Antimicrobial Proteins, 6, 123—135.

38. O’Toole, G. A., Pratt, L. A., Watnick, P. I., Newman, D. K., Weaver, V. B., Kolter, R., 1999: Genetic approaches to study of biofilms. Methods Enzymol., 310, 91—109.

39. Pompilio, A., Crocetta, V., Confalone, P., Nicoletti, M., Petrucca, A., Guarnieri, S. et al., 2010: Adhesion to and biofilm formation on IB3-1 bronchial cells by Stenotrophomonas maltophilia isolates from cystic fibrosis patients. BMC Microbiol., 10, 102—105.

40. Rodrigues, K. L., Caputo, L. R., Carvalho, J. C., Evangelista, J., Schneedorf, J. M., 2005: Antimicrobial and healing activity of kefir and kefiran extract. International Journal of Antimicrobial Agents, 25, 404—408.

41. Ross, P. R., Morgan, S., Hill, C., 2002: Preservation and fermentation: past, present and future. Int. J. Food Microbiol., 79, 3—16.

42. Slížová, M., Nemcová, R., Maďar, M., Hádryová, J., Gancarčíková, S., Popper, M., Pistl, J., 2015: Analysis of biofilm formation by intestinal lactobacilli. Can. J. Microbiol., 61, 437—446.

43. Terraf, M. C., Juárez Tomás, M. S., Nader-Macías, M. E., Silva, C., 2012: Screening of biofilm formation by beneficial vaginal lactobacilli and influence of culture media components. J. Appl. Microbiol., 113, 1517—29.

44. Urdaneta, E., Barrenetxe, J., Aranguren, P., Irigoyen, A., Marzo, F., Ibanez, F. C., 2007: Intestinal beneficial effects of kefir-supplemented diet in rats. Nutr. Res., 27, 653—658.

45. Vesterlund, S., Vankerckhoven, V., Saxelin, M., Goossens, H., Salminen, S., Ouwehand, A. C., 2007: Safety assessment of Lactobacillus strains: presence of putative risk factors in faecal, blood and probiotic isolates. Int. J. Food Microbiol., 116, 325—331.

46. Zheng, Y., Lu, Y., Wang, J., Yang, L., Pan, C., Huang, Y., 2013: Probiotic properties of Lactobacillus strains isolated from Tibetan kefir grains. PLoS ONE, 8, e69868.

Journal Information

Metrics

All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 406 398 31
PDF Downloads 138 137 14