International Journal of Nutrition and Food Sciences

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Flow Cytometric Assessment of Lactococcus Lactis Isolates Viability after Lyophilization

Received: 11 January 2014    Accepted: 20 August 2014    Published: 30 August 2014
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Abstract

Lactococcus lactis strains tolerance to the lyophilization process is one of the criteria used during the selection of starter cultures in cheese production. The viability of L. lactis strains isolated from silage and milk after the lyophilization process was evaluated in the present study by flow cytometry and cell plate counting. The LIVE/DEAD BacLightTM kit, which contains the dyes SYTO 9 and propidium iodide, was used to stain the cells for flow cytometry analysis. The dried cells were also plated on M17 agar and incubated at 30 °C for 24 hours. The strain L. lactis LBU.1, isolated from buffalo milk, showed the highest tolerance to the lyophilization process, according to flow cytometry and cell plate counting. Although the strains of L. lactis isolated from silage were not the most resistant to the lyophilization process, it is noteworthy that, compared to strains isolated from conventional sources such as cow and goat milk, L. lactis strains from silage showed higher tolerance to the lyophilization process.

DOI 10.11648/j.ijnfs.20140305.15
Published in International Journal of Nutrition and Food Sciences (Volume 3, Issue 5, September 2014)
Page(s) 391-396
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Lactococcus Lactis, Lyophilization, Viability, Flow Cytometry

References
[1] Coulibaly, I., Dubois-Dauphin, R., Destain, J., Fauconnier, M., Lognay, G., & Thonart, P. (2010). The resistance to freeze-drying and to storage was determined as the cellular ability to recover its survival rate and acidification activity. International Journal of Microbiology 1-9.
[2] Selwal, K., Selwal, M., & Gandhi, D. N.(2011). Effect of freeze drying process on some properties of Streptococcus thermophilus. Brazilian Journal of Microbiology 42: 1500-1505.
[3] Strasser, S., Neureiter, M., Geppl, M., Braun, R., & Danner, H.(2009). Influence of lyophilization, fluidized bed drying, addition of protectants, and storage on the viability of lactic acid bacteria. Journal of Applied Microbiology 107: 167-177.
[4] Pyar, H., & Peh, K.(2011). Effect cryoprotective agents on survival and stability of Lactobacillus acidophilus cultured in food-grade medium. International Journal of Dairy Technology 64: 578-584.
[5] Simões, C., Alakomi, H., Maukonen, J., & Saarela, M.(2010). Expression of clpL1 and clpL2 genes in Lactobacillus rhamnosus VTT E-97800 after exposure to acid and heat stress treatments or to freeze-drying. Beneficial Microbes 1: 253-257.
[6] Bouix, M., & Ghorbal, S. (2012). Rapid enumeration of Oenococcus oeni during malolactic fermentation by flow cytometry. Journal of Applied Microbiology 1-7.
[7] Muller, J. A. Stanton, C., Sybesma, W., Fitzgerald, G. F., & Ross, R. P.(2010). Reconstitution conditions for dried probiotic powders represent a critical step in determining cell viability. Journal of Applied Microbiology 108: 1369-1379.
[8] Soejima, T., Minami, J., & Iwatsuki, K.(2012). The exclusive use of flow cytometry to evaluate the antibiotic-susceptibility. Biochimica et Biophysica Acta 1820: 1980-1986.
[9] Paparella, A., Taccogna, L., Aguzzi, I., Chaves-López, C., Serio, A., Marsilio, F., & Suzzi, G.(2008). Flow cytometric assessment of the antimicrobial activity of essential oils against Listeria monocytogenes. Food Control 19: 1174-1182.
[10] Michal, K., Baszczynski, M., Lehnert, R., Mota, A., Teixeira, J. A., & Brányik, T. (2009). Flow cytometry for age assessment of a yeast population and its application in beer fermentations. Journal of the Institute of Brewing 115: 253-258.
[11] Xu, J., Zhou, F., Ji, B. P., Pei, R. S., & Xu, N.(2008). The antibacterial mechanism of carvacrol and thymol against Escherichia coli. Letters in Applied Microbiology 47: 174-179.
[12] Marchetti, A., Lelong, E., & Cosson, P. (2009).A measure of endosomal pH by flow cytometry in Dictyostelium. BMC Research Notes 2-7.
[13] Kramer, M., Obermajer, N., Matijasic, B. B., Rogelj, I., & Kmetec, V. (2009). Quantification of live and dead probiotic bacteria in lyophilized product by real-time PCR and by flow cytometry. Applied Microbiology Biotechnology 84:1137-1147.
[14] Boulos, L., Prévost, M., Barbeau, B., Coallier, J., & Desjardins, R. (1999). Live/Dead® BacLightTM: application of a new rapid staining method for direct enumeration of viable and total bacteria in drinking water. Journal Microbiology Methods 37: 77-86.
[15] Nomura, M., Kobayashi, M., Narita, T., Kimoto-Nira, H., & Okamoto, T.(2006). Phenotypic and molecular characterization of Lactococcus lactis from milk and plants. Journal of Applied Microbiology 101: 396-405.
[16] Morton, R. D. Aerobic plate count. In F. P. Downes, K. Ito (Eds.),(2001). Compendium of methods for the microbiological examination of foods (4th ed.), (pp. 1183-1193). Washington D.C.: American Public Health Association.
[17] Novo, D. J., Perlmutter, N. G., Hunt, R. H., & Shapiro, H. M.(2000). Multiparameter flow cytometric analysis of antibiotic effects on membrane potential, membrane permeability, and bacterial counts of Staphylococcus aureus and Micrococcus luteus. Antimicrobial Agents and Chemotherapy 44: 827-834.
[18] Morgan, C. A., Herman, N., White, P. A., & Vesey, G.(2006). Preservation of micro-organisms by drying: A review. Journal of Microbiology Methods 66:183-193
[19] Papadimitriou, K., Pratsinis, H., Nebe-von-Caron, G., Kletsas, D., & Tsakalidou, E.(2006). Rapid assessment of the physiological status of Streptococcus macedonicus by flow cytometry and fluorescence probes. International Journal of Food Microbiology 111: 197-205.
[20] Bouix, M., & Leveau, J. Y. (2001). Rapid assessment of yeast viability and yeast vitality during alcoholic fermentation . Journal of the Institute of Brewing 107: 217-225.
[21] Bunthof, C. J. & Abee, T. (2002). Development of a flow cytometric method to analyze subpopulations of bacteria in probiotic products and dairy starters. Applied and Environmental Microbiology 68: 2934-2942.
[22] Rault, A., Béal, C., Ghorbal, S., Ogier, J., & Bouix, M. (2007). Multiparametric flow cytometry allows rapid assessment and comparison of lactic acid bacteria viability after freeze and during frozen storage. Cryobiology 55: 35-43.
[23] Amor, K. B., Breeuwer, P., Verbaarschot, P., Rombouts, F. M., Akkermans, A. D., Vos, W. M., & Abee, T. (2002). Multiparametric flow cytometry and cell sorting for the assessment of viable, injured, and dead Bifidobacterium cells during Bile Salt Stress. Applied and Environmental Microbiology 68: 5209-5216.
[24] Bunthof, C. J., Braak, V., Breeuwer, P. Rombouts, F. M., & Abee, T. (1999). Rapid fluorescence assessment of the viability of stressed Lactococcus lactis. Applied and Environmental Microbiology 65: 3681-3689.
Author Information
  • Departamento de Microbiologia, Universidade Federal de Vi?osa, Vi?osa, Brazil

  • Departamento de Medicina Veterinária Preventiva, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

  • Departamento de Zootecnia, Universidade Federal de Vi?osa, Vi?osa, Brazil

  • Departamento de Microbiologia, Universidade Federal de Vi?osa, Vi?osa, Brazil

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    Eliana dos Santos Leandro, Graciela Kunrath Lima, Antônio Fernandes de Carvalho, Odilon Gomes Pereira, Célia Alencar de Moraes. (2014). Flow Cytometric Assessment of Lactococcus Lactis Isolates Viability after Lyophilization. International Journal of Nutrition and Food Sciences, 3(5), 391-396. https://doi.org/10.11648/j.ijnfs.20140305.15

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    ACS Style

    Eliana dos Santos Leandro; Graciela Kunrath Lima; Antônio Fernandes de Carvalho; Odilon Gomes Pereira; Célia Alencar de Moraes. Flow Cytometric Assessment of Lactococcus Lactis Isolates Viability after Lyophilization. Int. J. Nutr. Food Sci. 2014, 3(5), 391-396. doi: 10.11648/j.ijnfs.20140305.15

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    AMA Style

    Eliana dos Santos Leandro, Graciela Kunrath Lima, Antônio Fernandes de Carvalho, Odilon Gomes Pereira, Célia Alencar de Moraes. Flow Cytometric Assessment of Lactococcus Lactis Isolates Viability after Lyophilization. Int J Nutr Food Sci. 2014;3(5):391-396. doi: 10.11648/j.ijnfs.20140305.15

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  • @article{10.11648/j.ijnfs.20140305.15,
      author = {Eliana dos Santos Leandro and Graciela Kunrath Lima and Antônio Fernandes de Carvalho and Odilon Gomes Pereira and Célia Alencar de Moraes},
      title = {Flow Cytometric Assessment of Lactococcus Lactis Isolates Viability after Lyophilization},
      journal = {International Journal of Nutrition and Food Sciences},
      volume = {3},
      number = {5},
      pages = {391-396},
      doi = {10.11648/j.ijnfs.20140305.15},
      url = {https://doi.org/10.11648/j.ijnfs.20140305.15},
      eprint = {https://download.sciencepg.com/pdf/10.11648.j.ijnfs.20140305.15},
      abstract = {Lactococcus lactis strains tolerance to the lyophilization process is one of the criteria used during the selection of starter cultures in cheese production. The viability of L. lactis strains isolated from silage and milk after the lyophilization process was evaluated in the present study by flow cytometry and cell plate counting. The LIVE/DEAD BacLightTM kit, which contains the dyes SYTO 9 and propidium iodide, was used to stain the cells for flow cytometry analysis. The dried cells were also plated on M17 agar and incubated at 30 °C for 24 hours. The strain L. lactis LBU.1, isolated from buffalo milk, showed the highest tolerance to the lyophilization process, according to flow cytometry and cell plate counting. Although the strains of L. lactis isolated from silage were not the most resistant to the lyophilization process, it is noteworthy that, compared to strains isolated from conventional sources such as cow and goat milk, L. lactis strains from silage showed higher tolerance to the lyophilization process.},
     year = {2014}
    }
    

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  • TY  - JOUR
    T1  - Flow Cytometric Assessment of Lactococcus Lactis Isolates Viability after Lyophilization
    AU  - Eliana dos Santos Leandro
    AU  - Graciela Kunrath Lima
    AU  - Antônio Fernandes de Carvalho
    AU  - Odilon Gomes Pereira
    AU  - Célia Alencar de Moraes
    Y1  - 2014/08/30
    PY  - 2014
    N1  - https://doi.org/10.11648/j.ijnfs.20140305.15
    DO  - 10.11648/j.ijnfs.20140305.15
    T2  - International Journal of Nutrition and Food Sciences
    JF  - International Journal of Nutrition and Food Sciences
    JO  - International Journal of Nutrition and Food Sciences
    SP  - 391
    EP  - 396
    PB  - Science Publishing Group
    SN  - 2327-2716
    UR  - https://doi.org/10.11648/j.ijnfs.20140305.15
    AB  - Lactococcus lactis strains tolerance to the lyophilization process is one of the criteria used during the selection of starter cultures in cheese production. The viability of L. lactis strains isolated from silage and milk after the lyophilization process was evaluated in the present study by flow cytometry and cell plate counting. The LIVE/DEAD BacLightTM kit, which contains the dyes SYTO 9 and propidium iodide, was used to stain the cells for flow cytometry analysis. The dried cells were also plated on M17 agar and incubated at 30 °C for 24 hours. The strain L. lactis LBU.1, isolated from buffalo milk, showed the highest tolerance to the lyophilization process, according to flow cytometry and cell plate counting. Although the strains of L. lactis isolated from silage were not the most resistant to the lyophilization process, it is noteworthy that, compared to strains isolated from conventional sources such as cow and goat milk, L. lactis strains from silage showed higher tolerance to the lyophilization process.
    VL  - 3
    IS  - 5
    ER  - 

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