In vitro digestion as a tool for functional isolation of a probiotic potential Lactobacillus rhamnosus

Authors

DOI:

https://doi.org/10.33448/rsd-v9i10.8544

Keywords:

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.

Published

27/09/2020

How to Cite

LIMA, M. dos S. F. de .; SOUZA, K. M. S. de .; PASTRANA, L. .; SOARES, M. T. C. V. .; PORTO, A. L. F. . In vitro digestion as a tool for functional isolation of a probiotic potential Lactobacillus rhamnosus . Research, Society and Development, [S. l.], v. 9, n. 10, p. e3119108544, 2020. DOI: 10.33448/rsd-v9i10.8544. Disponível em: https://www.rsdjournal.org/index.php/rsd/article/view/8544. Acesso em: 8 aug. 2022.

Issue

Section

Agrarian and Biological Sciences