DOI QR코드

DOI QR Code

Screening of Probiotic Activities of Lactobacilli Strains Isolated from Traditional Tibetan Qula, A Raw Yak Milk Cheese

  • Zhang, Bei (Henan Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University) ;
  • Wang, Yanping (Henan Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University) ;
  • Tan, Zhongfang (Henan Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University) ;
  • Li, Zongwei (Henan Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University) ;
  • Jiao, Zhen (Henan Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University) ;
  • Huang, Qunce (Henan Provincial Key Laboratory of Ion Beam Bio-engineering, Zhengzhou University)
  • Received : 2015.10.14
  • Accepted : 2016.01.14
  • Published : 2016.10.01

Abstract

In this study, 69 lactobacilli isolated from Tibetan Qula, a raw yak milk cheese, were screened for their potential use as probiotics. The isolates were tested in terms of: Their ability to survive at pH 2.0, pH 3.0, and in the presence of 0.3% bile salts; tolerance of simulated gastric and intestinal juices; antimicrobial activity; sensitivity against 11 specific antibiotics; and their cell surface hydrophobicity. The results show that out of the 69 strains, 29 strains (42%) had survival rates above 90% after 2 h of incubation at pH values of 2.0 or 3.0. Of these 29 strains, 21 strains showed a tolerance for 0.3% bile salt. Incubation of these 21 isolates in simulated gastrointestinal fluid for 3 h revealed survival rates above 90%; the survival rate for 20 of these isolates remained above 90% after 4 h of incubation in simulated intestinal fluid. The viable counts of bacteria after incubation in simulated gastric fluid for 3 h and simulated intestinal fluid for 4 h were both significantly different compared with the counts at 0 h (p<0.001). Further screening performed on the above 20 isolates indicated that all 20 lactobacilli strains exhibited inhibitory activity against Micrococcus luteus ATCC 4698, Bacillus subtilis ATCC 6633, Listeria monocytogenes ATCC 19115, and Salmonella enterica ATCC 43971. Moreover, all of the strains were resistant to vancomycin and streptomycin. Of the 20 strains, three were resistant to all 11 elected antibiotics (ciprofloxacin, erythromycin, tetracycline, penicillin G, ampicillin, streptomycin, polymyxin B, vancomycin, chloramphenicol, rifampicin, and gentamicin) in this study, and five were sensitive to more than half of the antibiotics. Additionally, the cell surface hydrophobicity of seven of the 20 lactobacilli strains was above 70%, including strains Lactobacillus casei 1,133 (92%), Lactobacillus plantarum 1086-1 (82%), Lactobacillus casei 1089 (81%), Lactobacillus casei 1138 (79%), Lactobacillus buchneri 1059 (78%), Lactobacillus plantarum 1141 (75%), and Lactobacillus plantarum 1197 (71%). Together, these results suggest that these seven strains are good probiotic candidates, and that tolerance against bile acid, simulated gastric and intestinal juices, antimicrobial activity, antibiotic resistance, and cell surface hydrophobicity could be adopted for preliminary screening of potentially probiotic lactobacilli.

Keywords

Probiotic Activities;Screening;Lactobacilli;Qula Cheese

Acknowledgement

Supported by : National Natural Science Foundation of China

References

  1. Dasen, A., F. Berthier, R. Grappin, A. G. Williams, and J. Banks. 2003. Genotypic and phenotypic characterization of the dynamics of the lactic acid bacterial population of adjunctcontaining Cheddar cheese manufactured from raw and microfiltered pasteurized milk. J. Appl. Microbiol. 94:595-607. https://doi.org/10.1046/j.1365-2672.2003.01878.x
  2. de Almeida Junior, W. L. G., Í. da Silva Ferrari, J. V. de Souza, C. D. A. da Silva, M. M. da Costa, and F. S. Dias. 2015. Characterization and evaluation of lactic acid bacteria isolated from goat milk. Food Control 53:96-103. https://doi.org/10.1016/j.foodcont.2015.01.013
  3. de Vos, W. M. 2011. Systems solutions by lactic acid bacteria:from paradigms to practice. Microb. Cell Fact. 10:S2. https://doi.org/10.1186/1475-2859-10-S1-S2
  4. Essid, I., M. Medini, and M. Hassouna. 2009. Technological and safety properties of Lactobacillus plantarum strains isolated from a Tunisian traditional salted meat. Meat Sci. 81:203-208. https://doi.org/10.1016/j.meatsci.2008.07.020
  5. Ennahar, S., T. Sashihara, K. Sonomoto, and A. Ishizaki. 2000. Class IIa bacteriocins: Biosynthesis, structure and activity. FEMS Microbiol. Rev. 24:85-106. https://doi.org/10.1111/j.1574-6976.2000.tb00534.x
  6. Gilliland, S. E., T. E. Staley, and L. J. Bush. 1984. Importance of bile tolerance of Lactobacillus acidophilus used as a dietary adjunct. J. Dairy Sci. 67:3045-3051. https://doi.org/10.3168/jds.S0022-0302(84)81670-7
  7. Holzapfel, W. H., P. Haberer, J. Snel, and U. Schillinger. 1998. Overview of gut flora and probiotics. Int. J. Food Microbiol. 41:85-101. https://doi.org/10.1016/S0168-1605(98)00044-0
  8. Hyronimus, B., C. Le Marrec, A. H. Sassi, and A. Deschamps. 2000. Acid and bile tolerance of spore-forming lactic acid bacteria. Int. J. Food Microbiol. 61:193-197. https://doi.org/10.1016/S0168-1605(00)00366-4
  9. Jacobsen, C. N., V. Rosenfeldt Nielsen, A. E. Hayford, P. L. Moller, K. F. Michaelsen, A. Paerregaard, B. Sandstrom, M. Tvede, and M. Jakobsen. 1999. Screening of probiotic activities of forty-seven strains of Lactobacillus spp. by in vitro techniques and evaluation of the colonization ability of five selected strains in humans. Appl. Environ. Microbiol. 65:4949-4956.
  10. Jacobsen, L., A. Wilcks, K. Hammer, G. Huys, D. Gevers, and S. R. Andersen. 2007. Horizontal transfer of tet(M) and erm(B) resistance plasmids from food strains of Lactobacillus plantarum to Enterococcus faecalis JH2-2 in the gastrointestinal tract of gnotobiotic rats. FEMS Microbiol. Ecol. 59:158-166. https://doi.org/10.1111/j.1574-6941.2006.00212.x
  11. Klare, I., C. Konstabel, G. Werner, G. Huys, V. Vankerckhoven, G. Kahlmeter, B. Hildebrandt, S. Muller-Bertling, W. Witte, and H. Goossens. 2007. Antimicrobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for probiotic or nutritional use. J. Antimicrob. Chemother. 59:900-912. https://doi.org/10.1093/jac/dkm035
  12. Chung, H. S., Y. B. Kim, S. L. Chun, and G. E. Ji. 1999. Screening and selection of acid and bile resistant bifidobacteria. Int. J. Food Microbiol. 47:25-32. https://doi.org/10.1016/S0168-1605(98)00180-9
  13. CLSI (Clinical and Laboratory Standards Institute). 2012. Performance Standards for Antimicrobial Susceptibility Testing. 22nd edn. Clinical and Laboratory Standards Institute, Wayne, PA, USA.
  14. Ammor, S., G. Tauveron, E. Dufour, and I. Chevallier. 2006. Antibacterial activity of lactic acid bacteria against spoilage and pathogenic bacteria isolated from the same meat smallscale facility: 1-Screening and characterization of the antibacterial compounds. Food Control 17:454-461. https://doi.org/10.1016/j.foodcont.2005.02.006
  15. Asurmendi, P., M. J. Garcia, L. Pascual, and L. Barberis. 2015. Biocontrol of Listeria monocytogenes by lactic acid bacteria isolated from brewer's grains used as feedstuff in Argentina. J. Stored Prod. Res. 61:27-31. https://doi.org/10.1016/j.jspr.2015.02.001
  16. Aymerich, M. T., M. Garriga, J. M. Monfort, I. Nes, and M. Hugas. 2000. Bacteriocin-producing lactobacilli in Spanish-style fermented sausages: characterization of bacteriocins. Food Microbiol. 17:33-45. https://doi.org/10.1006/fmic.1999.0275
  17. Bao, Y., Y. Zhang, Y. Zhang, Y. Liu, S. Wang, X. Dong, Y. Wang, and H. Zhang. 2010. Screening of potential probiotic properties of Lactobacillus fermentum isolated from traditional dairy products. Food Control 21:695-701. https://doi.org/10.1016/j.foodcont.2009.10.010
  18. Charteris, W. P., P. M. Kelly, L. Morelli, and J. K. Collins. 1998. Antibiotic susceptibility of potentially probiotic Lactobacillus species. J. Food Prot. 61:1636-1643. https://doi.org/10.4315/0362-028X-61.12.1636
  19. Charteris, W. P., P. M. Kelly, L. Morelli, and J. K. Collins. 1998. Development and application of an in vitro methodology to determine the transit tolerance of potentially probiotic Lactobacillus and Bifidobacterium species in the upper human gastrointestinal tract. J. Appl. Microbiol. 84:759-768. https://doi.org/10.1046/j.1365-2672.1998.00407.x
  20. Mathara, J. M., U. Schillinger, P. M. Kutima, S. K. Mbugua, C. Guigas, C. Franz, and W. H. Holzapfel. 2008. Functional properties of Lactobacillus plantarum strains isolated from Maasai traditional fermented milk products in Kenya. Curr. Microbiol. 56:315-321. https://doi.org/10.1007/s00284-007-9084-6
  21. Musikasang, H., A. Tani, A. H-kittikun, and S. Maneerat. 2009. Probiotic potential of lactic acid bacteria isolated from chicken gastrointestinal digestive tract. World J. Microbiol. Biotechnol. 25:1337-1345. https://doi.org/10.1007/s11274-009-0020-8
  22. Nostro, A., M. A. Cannatelli, G. Crisafi, A. D. Musolino, F. Procopio, and V. Alonzo. 2004. Modifications of hydrophobicity, in vitro adherence and cellular aggregation of Streptococcus mutans by Helichrysum italicum extract. Lett. Appl. Microbiol. 38:423-427. https://doi.org/10.1111/j.1472-765X.2004.01509.x
  23. Ouoba, L. I. I., V. Lei, and L. B. Jensen. 2008. Resistance of potential probiotic lactic acid bacteria and bifidobacteria of African and European origin to antimicrobials: Determination and transferability of the resistance genes to other bacteria. Int. J. Food Microbiol. 121:217-224. https://doi.org/10.1016/j.ijfoodmicro.2007.11.018
  24. Prasad, J., H. Gill, J. Smart, and P. K. Gopal. 1998. Selection and characterisation of Lactobacillus and Bifidobacterium strains for use as probiotics. Int. Dairy. J. 8:993-1002. https://doi.org/10.1016/S0958-6946(99)00024-2
  25. Rojo-Bezares, B., Y. Saenz, P. Poeta, M. Zarazaga, F. Ruiz-Larrea, and C. Torres. 2006. Assessment of antibiotic susceptibility within lactic acid bacteria strains isolated from wine. Int. J. Food Microbiol. 111:234-240. https://doi.org/10.1016/j.ijfoodmicro.2006.06.007
  26. Rosenberg, M., D. Gutnick, and E. Rosenberg. 1980. Adherence of bacteria to hydrocarbons: A simple method for measuring cellsurface hydrophobicity. FEMS Microbiol. Lett. 9:29-33. https://doi.org/10.1111/j.1574-6968.1980.tb05599.x
  27. Solieri, L., A. Bianchi, G. Mottolese, F. Lemmetti, and P. Giudici. 2014. Tailoring the probiotic potential of non-starter Lactobacillus strains from ripened Parmigiano Reggiano cheese by in vitro screening and principal component analysis. Food Microbiol. 38:240-249. https://doi.org/10.1016/j.fm.2013.10.003
  28. Tan, Z., H. Pang, Y. Duan, G. Qin, and Y. Cai. 2010. 16S ribosomal DNA analysis and characterization of lactic acid bacteria associated with traditional Tibetan Qula cheese made from yak milk. Anim. Sci. J. 81:706-713. https://doi.org/10.1111/j.1740-0929.2010.00785.x
  29. Temmerman, R., B. Pot, G. Huys, and J. Swings. 2003. Identification and antibiotic susceptibility of bacterial isolates from probiotic products. Int. J. Food Microbiol. 81:1-10. https://doi.org/10.1016/S0168-1605(02)00162-9
  30. Tulini, F. L., L. K. Winkelstroter, and E. C. P. De Martinis. 2013. Identification and evaluation of the probiotic potential of Lactobacillus paraplantarum FT259, a bacteriocinogenic strain isolated from Brazilian semi-hard artisanal cheese. Anaerobe 22:57-63. https://doi.org/10.1016/j.anaerobe.2013.06.006
  31. Tulumoglu, S., Z. N. Yuksekdag, Y. Beyatli, O. Simsek, B. Cinar, and E. Yasar. 2013. Probiotic properties of lactobacilli species isolated from children's feces. Anaerobe 24:36-42. https://doi.org/10.1016/j.anaerobe.2013.09.006
  32. Wadstroum, T., K. Andersson, M. Sydow, L. Axelsson, S. Lindgren, and B. Gullmar. 1987. Surface properties of lactobacilli isolated from the small intestine of pigs. J. Appl. Microbiol. 62:513-520.
  33. Xanthopoulos, V., E. Litopoulou-Tzanetaki, and N. Tzanetakis. 2000. Characterization of Lactobacillus isolates from infant faeces as dietary adjuncts. Food Microbiol. 17:205-215. https://doi.org/10.1006/fmic.1999.0300
  34. Zhang, L. 2011. Evaluation of the Potential Probiotic Properties and Immune Regulation Function of Lactobacillus Strains Isolated from Traditional Fermented Yak Milk. Ph.D. Thesis, Gansu Agricultural University, Lanzhou, Gansu, China.

Cited by

  1. Characterization of the Antimicrobial Activity of Lactic Acid Bacteria Isolated from Buffalo Milk in West Sumatera (Indonesia) Against Listeria monocytogenes vol.16, pp.8, 2017, https://doi.org/10.3923/pjn.2017.645.650
  2. Lactic Acid Bacteria from Kefir Increase Cytotoxicity of Natural Killer Cells to Tumor Cells vol.7, pp.4, 2018, https://doi.org/10.3390/foods7040048
  3. Functional and safety characterization of autochthonous Lactobacillus paracasei FS103 isolated from sheep cheese and its survival in sheep and cow fermented milks during cold storage vol.69, pp.2, 2019, https://doi.org/10.1007/s13213-018-1416-1