In vitro digestion as a tool for functional isolation of a probiotic potential Lactobacillus rhamnosus
DOI:
https://doi.org/10.33448/rsd-v9i10.8544Keywords:
Kefir; Sheep milk; In vitro digestion; Lactobacillus rhamnosus.Abstract
This study set out to isolate microrganisms strains with probiotic characteristics after simulating the in vitro digestion of sheep milk fermented by kefir grains. Three lactobacilli with probiotic characteristics were isolated and identified as Lactobacillus rhamnosus. Assays characterized these strains as probiotic since they tolerated acid pH and bile salts, had antibiotic resistance, antagonist activity, antioxidant activity, presence of β-galactosidase enzyme and other tests revealed adhesion capacity. All strains presented antioxidant activity and survived at different pH and bile salts. These strains can be considered safe because they were susceptible to antibiotics tested, possess antagonist activity to pathogens and high β-galactosidase activity. As to adhesion criteria (hydrophobicity and autoaggregation), L. rhamnosus Lb16 stood out, as it also adheres to the intestinal epithelium cells of mice. The analysis of L. rhamnosus Lb16 can assist the dairy industry to enhance the potential human health benefits of its products. This paper is an important contribution to probiotics isolated after simulation of the in vitro digestion of fermented sheep milk by kefir grains, this has a differential due to its different characteristics which afforded the isolation of resistant strains to gastrointestinal conditions.
References
Archer, A .C., & Halami, P. M. (2015). Probiotic attributes of Lactobacillus fermentum isolated from human feces and dairy products. Applied Microbiology and Biotechnology 99(19): 8113–8123. doi: 10.1007/s00253-015-6679-x
Clinical and Laboratory Standars Institute - CLSI. (2012). Performance standards for antimicrobial disk susceptibility tests. Pensnylvania: CLSI.
Dertli, E., & Çon, A. H. (2017). Microbial diversity of traditional kefir grains and their role on kefir aroma. LWT- Food Science and Technology 85(Part A): 151-157. doi: 10.1016/j.lwt.2017.07.017
Ghasemi-Sadabadi, M., Ebrahimnezhad, Y., Shaddel-Tili, A., Bannapour-Ghaffari, V., Kozehgari, H. & Didehvar, M. (2019). The effects of fermented milk products (kefir and yogurt) and probiotic on performance, carcass characteristics, blood parameters, and gut microbial population in broiler chickens. Archives Animal Breeding 62(1): 361–374. doi: 10.5194/aab-62-361-2019
Gil-Rodríguez, A. M., Carrascosa, A. V., & Requena, T. (2015). Yeasts in foods and beverages: In vitro characterization of probiotic traits. LWT - Food Science and Technology 64(2): 1156–1162. doi: 10.1016/j.lwt.2015.07.042
Gul, O., Atalar, I., Mortas, M., & Dervisoglu, M. (2018). Rheological, textural, colour and sensorial properties of kefir produced with buffalo milk using kefir grains and starter culture: A comparison with cows’ milk kefir. International Journal of Dairy Technology 71(S1): 73-80. doi: 10.1111/1471-0307.12503
Izquierdo-González, J. J., Amil-Ruiz, F., Zazzu, S., Sánchez-Lucas, R., Fuentes-Almagro, C. A. & Rodríguez-Ortega, M. J. (2019). Proteomic analysis of goat milk kefir: Profiling the fermentation-time dependent protein digestion and identification of potential peptides with biological activity. Food chemistry 295: 456-465. doi: 10.1016/j.foodchem.2019.05.178.
Jamaly, N., Benjouad, A., & Bouksaim, M. (2011). Probiotic Potential of Lactobacillus strains Isolated from Known Popular Traditional Moroccan Dairy Products. British Microbiology Research Journal 1(4): 79–94.
Jena, P. K., Trivedi, D., Thakore, K., Chaudhary, H., Giri, S. S., & Seshadri, S. (2013). Isolation and characterization of probiotic properties of lactobacilli isolated from rat fecal microbiota. Microbiology and Immunology 57(6): 407–416. doi: 10.1111/1348-0421.12054.
Kos, B., Šušković, J., Vuković, S., Sǐmpraga, M., Frece, J., & Matošić, S. (2003). Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. Journal of Applied Microbiology 94(6): 981–987. doi: 10.1046/j.1365-2672.2003.01915.x.
Kotzamanidis, C., Kourelis, A., Litopoulou-Tzanetaki, E., Tzanetakis, N. & Yiangou, M. (2010). Evaluation of adhesion capacity, cell surface traits and immunomodulatory activity of presumptive probiotic Lactobacillus strains. International Journal of Food Microbiology 140(2–3): 154–163. doi: 10.1016/j.ijfoodmicro.2010.04.004
Kumar, A., & Kumar, D. (2015). Characterization of Lactobacillus isolated from dairy samples for probiotic properties. Anaerobe 33: 117–123. doi: 10.1016/j.anaerobe.2015.03.004
Lima, M. S. F., Silva, R. A., Silva, M. F., Silva, P. A. B., Costa, R. M. P. B., Teixeira, J. A. C., Porto, A. L. F., & Cavalcanti, M. T. H. (2018). Brazilian Kefir-Fermented Sheep’s Milk, a Source of Antimicrobial and Antioxidant Peptides. Probiotic and Antimicrobial Proteins 10(3): 446-455. doi: 10.1007/s12602-017-9365-8.
Lima, M. S. F., Souza, K. M. S., Albuquerque, W. W. C., Teixeira, J. A. C., Cavalcanti, M. T. H., & Porto, A. L. F. (2017). Saccharomyces cerevisiae from Brazilian kefir-fermented milk: An in vitro evaluation of probiotic properties. Microbial Pathogenesis 10: 670-677. doi: 10.1016/j.micpath.2017.05.010
Meira, S. M. M., Helfer, V. E., Velho, R. V., Lopes, F. C. & Brandelli, A. (2012). Probiotic potential of Lactobacillus spp. isolated from Brazilian regional ovine cheese. Journal of Dairy Research 79(1): 119–27. doi: 10.1017/S0022029911000884.
Oliveira, A. P., Santos, G. A., Nomura, C. S., & Naozuka, J. (2019). Elemental chemical composition of products derived from kefir fermented milk. Journal of Food Composition and Analysis 78:86-90. doi: 10.1016/j.jfca.2019.02.005
Pringsulaka, O., Rueangyotchanthana, K., Suwannasai, N., Watanapokasin, R., Amnueysit, P., Sunthornthummas, S., Sukkhum, S., Sarawaneeyaruk, S. & Rangsiruji, A. (2015). In vitro screening of lactic acid bacteria for multi-strain probiotics. Livestock Science 174: 66–73. doi: 10.1016/j.livsci.2015.01.016
Ren, D., Li, C., Qin, Y., Yin, R., Du, S., Ye, F., Liu, C., Liu, H., Wang, M., Li, Y., Sun, Y., Li, X., Tian, M. & Jin, N. (2014). In vitro evaluation of the probiotic and functional potential of Lactobacillus strains isolated from fermented food and human intestine. Anaerobe 30:1–10. doi: 10.1016/j.anaerobe.2014.07.004
Rosenberg, M., Gutnick, D. & Rosenberg, E. (1980). Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiology Letters 9: 29–33. doi: 10.1111/j.1574-6968.1980.tb05599.x
Santos, A., San, M. M., Sanchez, A., Torres, J. M. & Marquina, D. (2003). The antimicrobial properties of different strains of Lactobacillus spp. isolated from kefir. Systematic and Applied Microbiology 26(3): 434–437. doi: 10.1078/072320203322497464
Savastano, M. L., Pati, S., Bevilacqua, A., Corbo, M. R., Rizzuti, A., Pischetsrieder, M. & Losito, I. (2020). Influence of the production technology on kefir characteristics: Evaluation of microbiological aspects and profiling of phosphopeptides by LC-ESI-QTOF-MS/MS. Food Research International 129: 108853. doi: 10.1016/j.foodres.2019.108853
Sumeri, I., Arike, L., Stekolštšikova, J., Uusna, R., Adamberg, S., Adamberg, K. & Paalme, T. (2010). Effect of stress pretreatment on survival of probiotic bacteria in gastrointestinal tract simulator. Applied Microbiology and Biotechnology 86(6): 1925–1931. doi: 10.1007/s00253-009-2429-2
Tang, W., Xing, Z., Li, C., Wang, J. & Wang, Y. (2017). Molecular mechanisms and in vitro antioxidant effects of Lactobacillus plantarum MA2. Food Chemistry 221: 1642–1649. doi: 10.1016/j.foodchem.2016.10.124
Tulini, F. L., Winkelströter, L. K. & De Martinis, E.C.P. (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. doi: 10.1016/j.anaerobe.2013.06.006
Tuo, Y., Zhang, W., Zhang, L., Ai, L., Zhang, Y., Han, X. & Yi, H. (2013). Study of probiotic potential of four wild Lactobacillus rhamnosus strain. Anaerobe 21: 22–27. doi: 10.1016/j.anaerobe.2013.03.007
Vimercati, W. C., Silva, C. A., Macedo, L. L., Fonseca, H. C., Guimarães, J. S., Abreu, L. R. & Pinto, S. M. (2020). Physicochemical, rheological, microbiological, and sensory properties of newly developed coffee flavored kefir. LWT - Food Science and Technology 123: 109069. doi: 10.1016/j.lwt.2020.109069
Vinderola, C.G. & Reinheimer, J.A. (2003). Lactic acid starter and probiotic bacteria: a comparative “in vitro” study of probiotic characteristics and biological barrier resistance. Food Research International 36(9–10): 895–904. doi: 10.1016/S0963-9969(03)00098-X
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. doi: 10.1371/journal.pone.0069868.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2020 Meire dos Santos Falcão de Lima; Karoline Mirella Soares de Souza; Lorenzo Pastrana; Maria Taciana Cavalcanti Vieira Soares; Ana Lúcia Figueiredo Porto
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
1) Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
2) Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
3) Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.
