Skip to main content Accessibility help
×
Home

Effect of freeze-drying on viability and in vitro probiotic properties of a mixture of lactic acid bacteria and yeasts isolated from kefir

  • Patricia A Bolla (a1) (a2), María de los Angeles Serradell (a2), Patricio J de Urraza (a2) and Graciela L De Antoni (a1) (a2)

Abstract

The effect of freeze-drying on viability and probiotic properties of a microbial mixture containing selected bacterial and yeast strains isolated from kefir grains (Lactobacillus kefir, Lactobacillus plantarum, Lactococcus lactis, Saccharomyces cerevisiae and Kluyveromyces marxianus) was studied. The microorganisms were selected according to their potentially probiotic properties in vitro already reported. Two types of formulations were performed, a microbial mixture (MM) suspended in milk and a milk product fermented with MM (FMM). To test the effect of storage on viability of microorganisms, MM and FMM were freeze-dried and maintained at 4°C for six months. After 180 days of storage at 4°C, freeze-dried MM showed better survival rates for each strain than freeze-dried FMM. The addition of sugars (trehalose or sucrose) did not improve the survival rates of any of the microorganisms after freeze-drying. Freeze-drying did not affect the capacity of MM to inhibit growth of Shigella sonnei in vitro, since the co-incubation of this pathogen with freeze-dried MM produced a decrease of 2 log in Shigella viability. The safety of freeze-dried MM was tested in mice and non-translocation of microorganisms to liver or spleen was observed in BALB/c mice feed ad libitum during 7 or 20 days. To our knowledge, this is the first report about the effect of freeze-drying on viability, in vitro probiotic properties and microbial translocation of a mixture containing different strains of both bacteria and yeasts isolated from kefir.

Copyright

Corresponding author

*For correspondence; e-mail: maserr@biol.unlp.edu.ar

References

Hide All
Abadias, M, Benabarre, A, Taixidó, N, Usall, J & Viñas, IV 2001 Effect of freeze drying and protectants on viability of the biocontrol yeast Candida sake. International Journal of Food Microbiology 65 173182
Abe, F, Muto, M, Yaeshima, T, Iwatsuki, K, Aihara, H, Ohashi, Y & Fujisawa, T 2009 Safety evaluation of probiotic bifidobacteria by analysis of mucin degradation activity and translocation ability. Anaerobe doi: 10.1016/j.anaerobe.2009.07.006
Abraham, A, De Antoni, G & Añón, M 1990 Effect of calcium on the cryopreservation of L. bulgaricus in different freezing media. Cryobiology 27 336
Angulo, L, Lopez, E & Lema, C 1993 Microflora present in kefir grains of the Galician region north-west of Spain. Journal of Dairy Research 60 263267
Balcazar, J, Vendrell, D, de Blas, I, Ruiz-Zarzuela, I & Muzquiz, J 2009 Effect of Lactococcus lactis CLFP 100 and Leuconostoc mesenteroides CLFP 196 on Aeromonas salmonicida Infection in brown trout (Salmo trutta). Journal of Molecular Microbiology and Biotechnology 17 153157
Berg, R 1999 Bacterial translocation from the gastrointestinal tract. Advances in Experimental Medical Biology 473 1130
Berny, J & Hennebert, G 1991 Viability and stability of yeast cells and filamentous fungus spore during freeze-drying: Effects of protectants and cooling rates. Mycologia 83 805815
Blanquet, S, Garrait, G, Beyssac, E, Parrier, C, Denis, S, Hébrard, G & Alric, M 2005 Effects of cryoprotectants on the viability and activity of freeze-dried recombinant yeast as novel oral drug delivery systems assessed by an artificial system. European Journal of Pharmacautics and Biopharmaceutics 61 3239
Bravo, D, Rodriguez, E & Medina, M 2009 Nisin and lacticin 481 coproduction by Lactococcus lactis strains isolated from raw ewes' milk. Journal of Dairy Science 92 48054811
Carvalho, A, Silva, J, Ho, P, Teixeira, P, Malcata, F & Gibbs, P 2002 Effect of additives on survival of freeze-dried Lactobacillus plantarum and Lactobacillus rhamnosus during storage. Biotechnology Letters 24 15871591
Carvalho, A, Silva, J, Ho, P & Teixeira, P 2004 Relevant factors for the preparation of freeze-dried lactic acid bacteria. International Dairy Journal 14 835847
Carvalho, A, Silva, J, Ho, P, Teixeira, P, Malcata, F & Gibbs, P 2003 Effect of various growth media upon survival during storage of freeze-dried Enterococcus faecalis and Enterococcus durans. Journal of Applied Microbiology 94 947952
Carvalho, A, Silva, J, Ho, P, Teixeira, P, Malcata, F & Gibbs, P 2004 Effects of various sugars added to growth and drying media upon thermotolerance and survival throughout storage of freeze-dried Lactobacillus delbrueckii ssp. bulgaricus. Biotechnology Progress 20 248254
Champagne, C, Gardner, N, Brochu, E & Beaulieu, Y 1991 The freeze-drying of lactic acid bacteria. A Review. Canadian Institute of Food Science and Technology 24 118128
De Antoni, G, Pérez, P, Abraham, A & Añón, C 1991 Trehalose, a cryoprotectant for L. bulgaricus. Cryobiology 26 149153
de Urraza, P & De Antoni, G 1997 Induced cryotolerance of Lactobacillus delbrueckii subsp. bulgaricus LBB by preincubation at suboptimal temperatures with a fermentable sugar. Cryobiology 35 159164
Echeverría, I, Garrote, G & De Antoni, G 2007 Formulación de una leche fermentada probiótica y con bajo contenido de lactosa. Tecnología Láctea Latinoamericana 48 4447
Farnworth, E 2005 Kefir – a complex probiotic. Food Science and Technology Bulletin 2 117
Garrote, G 2000 Bacterias lácticas y levaduras para la industria alimentaria: kefir. PhD Thesis Universidad Nacional de La Plata, Argentina
Garrote, G, Abraham, A & De Antoni, G 2001 Chemical and microbiological characterisation of kefir grains. Journal of Dairy Research 68 639652
Garrote, G, Delfederico, L, Bibiloni, R, Abraham, A, Perez, P, Semorile, L & De Antoni, G 2004 Lactobacilli isolated from kefir grains: evidence of the presence of S-layer proteins. Journal of Dairy Research 71 222230
Golowczyc, M, Gugliada, M, Hollmann, A, Delfederico, L, Garrote, G, Abraham, A, Semorile, L & De Antoni, G 2008 Characterization of homofermentative lactobacilli isolated from kefir grains: potential use as probiotic. Journal of Dairy Research 75 211217
Golowczyc, M, Mobili, P, Garrote, G, Abraham, A & De Antoni, G 2007 Protective action of Lactobacillus kefir carrying S-layer protein against Salmonella enterica serovar Enteritidis. International Journal of Food Microbiology 118 264273
Hugo, A, Kakisu, E, De Antoni, G & Perez, P 2008 Lactobacilli antagonize biological effects of enterohaemorrhagic Escherichia coli in vitro. Letters in Applied Microbiology 46 613619
Kunz, A, Noel, J & Firchok, M 2004 Two case of Lactobacillus bacteremia during probiotic treatment of short gut syndrome. Journal of Pediatric Gastroenterology and Nutrition 38 457458
Land, M, Rouster-Stevens, K, Woods, C, Cannon, M, Cnota, J & Shetty, A 2005 Lactobacillus sepsis associated with probiotic therapy. Pediatrics 115 178181
Leslie, S, Israeli, E, Lighthart, B, Crowe, J & Crowe, L 1995 Trehalose and sucrose protect both membranes and proteins in intact bacteria during drying. Applied and Environmental Microbiology 61 35923597
Linders, L, Wolkers, W, Hoekstra, F & van‘t Riet, K 1997 Effect of added carbohydrates on membrane phase behavior and survival of dried Lactobacillus plantarum. Cryobiology 35 3140
Lodato, P, Segovia de Huergo, M & Buera, M 1999 Viability and thermal stability of a strain of Saccharomyces cerevisiae freeze-dried in different sugar and polymer matrices. Applied Microbiology and Biotechnology 52 215220
Nyanga, L, Nout, M, Gadaga, T, Theelen, B, Boekhout, T & Zwietering, M 2007 Yeasts and lactic acid bacteria microbiota from masau (Ziziphus mauritiana) fruits and their fermented fruit pulp in Zimbabwe. International Journal of Food Microbiology 120 159166
Morgan, C, Herman, N, White, P & Vesey, G 2006 Preservation of microorganisms by drying; a review. Journal of Microbiological Methods 66 183193
Mugula, J, Narvhus, J & Sørhaug, T 2003 Use of starter cultures of lactic acid bacteria and yeasts in the preparation of togwa, a Tanzanian fermented food. International Journal of Food Microbiology 83 307318
Otero, M, Espeche, M & Nacer Macías, M 2007 Optimization of the freeze-drying and survival throughout storage of fFreeze-dried Lactobacillus gasseri and Lactobacillus delbrueckii subsp. delbrueckii for veterinarian probiotic applications. Process Biochemistry 42 14061411
Pavan, S, Desreumaux, P & Mercenier, A 2003 Use of mouse models to evaluate the persistence, safety, and immune modulation capacities of lactic acid bacteria. Clinical and Diagnostic Laboratory Immunology 10 696701
Romanin, D, Serradell, M, González-Maciel, D, Lausada, N, Garrote, G & Rumbo, M 2010 Down-regulation of intestinal epithelial innate response by probiotic yeast isolated from kefir. International Journal of Food Microbiology 140 102108.
Rossland, E, Andersen Borge, G, Langsrud, T & Sorhaug, T 2003 Inhibition of Bacillus cereus by strains of Lactobacillus and Lactococcus in milk. International Journal of Food Microbiology 89 205212
Sanders, M & Klaenhammer, T 2001 Invited review: the scientific basis of Lactobacillus acidophilus NCFM functionality as a probiotic. Journal of Dairy Science 84 319331
Selmer-Olsen, E, Birkeland, S & Sorhaug, T 1999 Effect of protective solutes on leakage from and survival of immobilized lactobacillus subjected to drying, storage and rehydration. Journal of Applied Microbiology 87 429437
Tsvetkov, T & Brankova, R 1983 Viability of micrococci and lactobacilli upon freezing and freeze-drying in the presence of different cryoprotectants. Cryobiology 20 318323
Vinderola, C, Duarte, J, Thangavel, D, Perdigon, G, Farnworth, E & Matar, C 2005 Immunomodulating capacity of kefir. Journal of Dairy Research 72 195202
Yakabe, T, Moore, E, Yokota, S, Sui, H, Nobuta, Y, Fukao, M, Palmer, H & Yajima, N 2009 Safety assessment of Lactobacillus brevis KB290 as a probiotic strain. Food and Chemical Toxicology 47 24502453
Zabala, V 2004 Caracterización de leches fermentadas obtenidas con gránulos de kefir: composición y propiedades probióticas y tecnológicas. Master Thesis, Universidad Nacional de La Plata, Argentina
Zhou, J, Shu, Q, Rutherfurd, K, Prasad, J, Gopal, P & Gill, H 2000 Acute oral toxicity and bacterial translocation studies on potentially probiotic strains of lactic acid bacteria. Food Chemical Toxicology 38 153161

Keywords

Effect of freeze-drying on viability and in vitro probiotic properties of a mixture of lactic acid bacteria and yeasts isolated from kefir

  • Patricia A Bolla (a1) (a2), María de los Angeles Serradell (a2), Patricio J de Urraza (a2) and Graciela L De Antoni (a1) (a2)

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed