In vitro evaluation of certain probiotic properties of lactic acid bacteria isolates
Article Sidebar
Main Article Content
Abstract
Bacteria carrying genes responsible for antibiotic resistance cannot be used in food production. For this reason,
exploring the antibiotic resistance profile of probiotic candidates and the antimicrobial substances they
produce are essential for probiotic strain selection. The aim of this study was to develop and evaluate additional
elements of a complex in vitro test system for rapid and efficient selection of a large number of putative
probiotic isolates. In a previous work, we had tested bacterial strains (n=217) isolated from Transylvanian raw
sheep milk, cultured sheep milk, and sheep cheese samples and we reduced the sample number to a total
of six Gram-positive, non-hemolytic, catalase-negative, well-aggregating, good acid and bile acid tolerating
strains. In this research, we investigated the antibiotic resistance and antimicrobial production capacity of the
pre-selected strains (n=6). The antimicrobial activity of the isolates was determined by the agar well diffusion
assay. Strains E15, E66, E173, E198, and E216 were found to inhibit the growth of both Salmonella Enteridis
ATCC 13076 and the control strain (i.e., Lactobacillus acidophilus ATCC 4356). Antibiotic resistance tests
were performed by the agar disk diffusion method. All six isolates belonging to the species of Levilactobacillus
brevis and Lactiplantibacillus plantarum were found to be resistant to several antibiotics and, therefore,
cannot be used for the manufacture of commercial probiotic products. In conclusion, our in vitro test system
proved to be capable of effectively screening out unsafe isolates.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
Fuller, R. (1989): Probiotics in man and animals. Journal of Applied Bacteriology 66 pp. 365-378. https://doi.org/10.1111/j.1365-2672.1989.tb05105.x
Hill, C., Guarner, F., Reid, G., Gibson, G.R., Merenstein, D.J., Pot, B., Morelli, L., Canani, R.B., Flint, H.J., Salminen, S., Calder, P.C., Sanders, M.E. (2014): The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nature Reviews Gastroenterology and Hepatology 11 pp. 506-514. https://doi.org/10.1038/nrgastro.2014.66
Fijan, S., Frauwallner, A., Varga, L., Langerholc, T., Rogelj, I., Lorber, M., Lewis, P., Povalej-Bržan, P. (2019): Health professionals’ knowledge of probiotics: an international survey. International Journal of Environmental Research and Public Health 16 3128. https://doi.org/10.3390/ijerph16173128
Fijan, S., ter Haar, J.A., Varga, L. (2021) Probiotic microorganisms and their benefit to human health. In Advances in Probiotics: Microorganisms in Food and Health, eds. Dhanasekaran, D., Sankaranarayanan, A. pp. 3-22. Academic Press and Elsevier: London, San Diego, Cambridge, Oxford. https://doi.org/10.1016/B978-0-12-822909-5.00001-0
Süle, J., Varga, L., Varga, K., Hatvan, Z., Kerényi, Z. (2022) Probiotikus baktériumtörzsek szelektálására alkalmas kísérleti rendszer egyes elemeinek kidolgozása (Developing basic elements of an experimental system for selection of probiotic bacterial strains). Magyar Állatorvosok Lapja 144 (közlésre benyújtva).
Papadimitriou, K., Zoumpopoulou, G., Foligné, B., Alexandraki, V., Kazou, M., Pot, B., Tsakalidou, E. (2015): Discovering probiotic microorganisms: in vitro, in vivo, genetic and omics approaches. Frontiers in Microbiology 6 58. https://doi.org/10.3389/fmicb.2015.00058
Williams, C.F., Walton, G.E., Jiang, L., Plummer, S., Garaiova, I., Gibson, G.R. (2015): Comparative analysis of intestinal tract models. Annual Review of Food Science and Technology 6 pp. 329-350. https://doi.org/10.1146/annurev-food-022814-015429
Antal, O., Némethné Szerdahelyi, E., Takács, K. (2020): In vitro humán emésztési modellek alkalmazása a táplálkozástudomány területén (Application of in vitro human digestion models in the field of nutrition science). Élelmiszervizsgálati Közlemények - Journal of Food Investigation 66 pp. 3141-3157.
Süle, J., Varga, L., Hatvan, Z., Kerényi, Z. (2022) In vitro tesztrendszer alkalmazása probiotikus baktériumtörzsek szelektálására (Application of an in vitro test system for the selection of probiotic bacterial strains). Élelmiszervizsgálati Közlemények - Journal of Food Investigation 68 pp. 3904-3927.
Ouwehand, A.C., Forssten, S., Hibberd, A.A., Lyra, A., Stahl, B. (2016): Probiotic approach to prevent antibiotic resistance. Annals of Medicine 48 pp. 246-255. https://doi.org/10.3109/07853890.2016.1 161232
Zheng, J.S., Wittouck, S., Salvetti, E., Franz, C.M.A.P., Harris, H.M.B., Mattarelli, P., O’Toole, P.W., Pot, B., Vandamme, P., Walter, J., Watanabe, K., Wuyts, S., Felis, G.E., Gänzle, M.G., Lebeer, S. (2020): A taxonomic note on the genus Lactobacillus: description of 23 novel genera, emended description of the genus Lactobacillus Beijerinck 1901, and union of Lactobacillaceae and Leuconostocaceae. International Journal of Systematic Evolutionary Microbiology 70 pp. 2782-2858. https://doi. org/10.1099/ijsem.0.004107
URL1: https://www.mn-net.com/media/pdf/91/f9/1f/Instruction-NucleoSpin-Microbial-DNA.pdf (Hozzáférés: 2022. 05. 03.)
URL2: https://blast.ncbi.nlm.nih.gov/Blast.cgi (Hozzáférés: 2022. 04. 11.)
Miao, J.Y., Guo, H.X., Ou, Y.W., Liu, G., Fang, X., Liao, Z.L., Ke, C., Chen, Y.J., Zhao, L.C., Cao, Y. (2014): Purification and characterization of bacteriocin F1, a novel bacteriocin produced by Lactobacillus paracasei subsp. tolerans FX-6 from Tibetan kefir, a traditional fermented milk from Tibet, China. Food Control 42 pp. 48-53. https://doi.org/10.1016/j.foodcont.2014.01.041
Maragkoudakis, P.A., Zoumpopoulou, G., Miaris, C., Kalantzopoulos, G., Pot, B., Tsakalidou, E. (2006): Probiotic potential of Lactobacillus strains isolated from dairy products. International Dairy Journal 16 pp. 189-199. https://doi.org/10.1016/j.idairyj.2005.02.009
International Organization for Standardization (ISO), International Dairy Federation (IDF) (2010): Milk and milk products – Determination of the minimal inhibitory concentration (MIC) of antibiotics applicable to bifidobacteria and non-enterococcal lactic acid bacteria (LAB). International Standard ISO 10932:2010(E) and IDF 223:2010(E). ISO, Geneva, Switzerland and IDF, Brussels, Belgium.
Huys, G., D’Haene, K., Swings, J. (2002): Influence of the culture medium on antibiotic susceptibility testing of food-associated lactic acid bacteria with the agar overlay disc diffusion method. Letters in Applied Microbiology 34 pp. 402-406. https://doi.org/10.1046/j.1472-765X.2002.01109.x
Charteris, W.P., Kelly, P.M., Morelli, L., Collins, J.K. (2001): Gradient diffusion antibiotic susceptibility testing of potentially probiotic lactobacilli. Journal of Food Protection 64 pp. 2007-2014. https://doi.org/10.4315/0362-028X-64.12.2007
Wolupeck, H.L., Morete, C.A., DallaSanta, O.R., Luciano, F.B., Madeira, H.M.F., de Macedo, R.E.F. (2017): Methods for the evaluation of antibiotic resistance in Lactobacillus isolated from fermented sausages. Ciência Rural 47 e20160966. https://doi.org/10.1590/0103-8478cr20160966