Characterization and Selection of <i>Lactobacillus</i> Strains with Potential Probiotic Applications

In the present study, four <i>Lactobacillus</i> strains (<i>Lacticaseibacillus casei</i> ATCC 393, <i>Lacticaseibacillus casei</i> RC-1, <i>Lactobacillus acidophilus</i> RC-2 and <i>Lacticaseibacillus rhamnosus</i> RC-10) were investigated...

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Main Authors: Yulian Tumbarski, Ivelina Peykova-Shapkova, Mihaela Ivanova, Remzi Cholakov, Agata Dutkiewicz, Krzysztof Grzymajło
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Applied Sciences
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Online Access:https://www.mdpi.com/2076-3417/15/6/2902
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author Yulian Tumbarski
Ivelina Peykova-Shapkova
Mihaela Ivanova
Remzi Cholakov
Agata Dutkiewicz
Krzysztof Grzymajło
author_facet Yulian Tumbarski
Ivelina Peykova-Shapkova
Mihaela Ivanova
Remzi Cholakov
Agata Dutkiewicz
Krzysztof Grzymajło
author_sort Yulian Tumbarski
collection DOAJ
description In the present study, four <i>Lactobacillus</i> strains (<i>Lacticaseibacillus casei</i> ATCC 393, <i>Lacticaseibacillus casei</i> RC-1, <i>Lactobacillus acidophilus</i> RC-2 and <i>Lacticaseibacillus rhamnosus</i> RC-10) were investigated to determine their probiotic potential. The strains were evaluated based on their resistance to simulated upper gastrointestinal conditions, tolerance to bile salts, antimicrobial activity, antibiotic resistance and ability to adhere to intestinal cells. All the strains were resistant to simulated upper gastrointestinal conditions (pH 2.5 + pepsin and pH 7.4 + pancreatin) and exhibited cell recovery rates varying between 74.24 and 87.50% (pH 2.5 + pepsin) and between 93.95 and 98.15% (pH 7.4 + pancreatin) at the 24th h of cultivation. The strains demonstrated resistance to various concentrations of bile salts (0.25, 0.5, 0.75 and 1%) with survival rates > 50% (<i>L. casei</i> ATCC 393 and <i>L. casei</i> RC-1) and >60% (<i>L. acidophilus</i> RC-2 and <i>L. rhamnosus</i> RC-10) in bile salt concentration of 0.25% at the 24th h of incubation. The four <i>Lactobacillus</i> components—<i>Lactobacillus</i> culture (LC), cell biomass (CB) and cell-free supernatant (CFS) exhibited moderate to high antimicrobial activity against six Gram-positive and six Gram-negative bacteria; however, they failed to inhibit the growth of yeasts and fungi tested in the antimicrobial screening. In addition, the neutralized to pH = 7.0 cell-free supernatant (NCFS) of the four strains completely lost its antimicrobial effect. The results for antibiotic susceptibility of four <i>Lactobacillus</i> strains showed that <i>L. casei</i> ATCC 393 was resistant to 11, <i>L. casei</i> RC-1 was resistant to 9, <i>L. acidophilus</i> RC-2 was resistant to 8, and <i>L. rhamnosus</i> RC-10 was resistant to 11 of the total 23 antibiotics tested. The highest degree of adhesion of the studied isolates to the intestinal cell line IPEC-J2 was observed in <i>L. casei</i> RC-1 (39.45%)<i>, L. rhamnosus</i> RC-10 (23.38%) and <i>L. acidophilus</i> RC-2 (19.25%) in comparison with the reference probiotic strain <i>L. casei</i> ATCC 393. Based on the results obtained, the strains studied can be considered as having probiotic potential.
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spelling doaj-art-36c29b99a91a4e0aa70da5ca1ca9b2fb2025-08-20T03:40:43ZengMDPI AGApplied Sciences2076-34172025-03-01156290210.3390/app15062902Characterization and Selection of <i>Lactobacillus</i> Strains with Potential Probiotic ApplicationsYulian Tumbarski0Ivelina Peykova-Shapkova1Mihaela Ivanova2Remzi Cholakov3Agata Dutkiewicz4Krzysztof Grzymajło5Department of Microbiology and Biotechnology, University of Food Technologies, 4002 Plovdiv, BulgariaDepartment of Milk and Dairy Products, University of Food Technologies, 4002 Plovdiv, BulgariaDepartment of Milk and Dairy Products, University of Food Technologies, 4002 Plovdiv, BulgariaÖzgazi B. V. Dairy Foods, 4879 AP Etten-Leur, The NetherlandsDepartment of Biochemistry and Molecular Biology, Wrocław University of Environmental and Life Sciences, 50-375 Wrocław, PolandDepartment of Biochemistry and Molecular Biology, Wrocław University of Environmental and Life Sciences, 50-375 Wrocław, PolandIn the present study, four <i>Lactobacillus</i> strains (<i>Lacticaseibacillus casei</i> ATCC 393, <i>Lacticaseibacillus casei</i> RC-1, <i>Lactobacillus acidophilus</i> RC-2 and <i>Lacticaseibacillus rhamnosus</i> RC-10) were investigated to determine their probiotic potential. The strains were evaluated based on their resistance to simulated upper gastrointestinal conditions, tolerance to bile salts, antimicrobial activity, antibiotic resistance and ability to adhere to intestinal cells. All the strains were resistant to simulated upper gastrointestinal conditions (pH 2.5 + pepsin and pH 7.4 + pancreatin) and exhibited cell recovery rates varying between 74.24 and 87.50% (pH 2.5 + pepsin) and between 93.95 and 98.15% (pH 7.4 + pancreatin) at the 24th h of cultivation. The strains demonstrated resistance to various concentrations of bile salts (0.25, 0.5, 0.75 and 1%) with survival rates > 50% (<i>L. casei</i> ATCC 393 and <i>L. casei</i> RC-1) and >60% (<i>L. acidophilus</i> RC-2 and <i>L. rhamnosus</i> RC-10) in bile salt concentration of 0.25% at the 24th h of incubation. The four <i>Lactobacillus</i> components—<i>Lactobacillus</i> culture (LC), cell biomass (CB) and cell-free supernatant (CFS) exhibited moderate to high antimicrobial activity against six Gram-positive and six Gram-negative bacteria; however, they failed to inhibit the growth of yeasts and fungi tested in the antimicrobial screening. In addition, the neutralized to pH = 7.0 cell-free supernatant (NCFS) of the four strains completely lost its antimicrobial effect. The results for antibiotic susceptibility of four <i>Lactobacillus</i> strains showed that <i>L. casei</i> ATCC 393 was resistant to 11, <i>L. casei</i> RC-1 was resistant to 9, <i>L. acidophilus</i> RC-2 was resistant to 8, and <i>L. rhamnosus</i> RC-10 was resistant to 11 of the total 23 antibiotics tested. The highest degree of adhesion of the studied isolates to the intestinal cell line IPEC-J2 was observed in <i>L. casei</i> RC-1 (39.45%)<i>, L. rhamnosus</i> RC-10 (23.38%) and <i>L. acidophilus</i> RC-2 (19.25%) in comparison with the reference probiotic strain <i>L. casei</i> ATCC 393. Based on the results obtained, the strains studied can be considered as having probiotic potential.https://www.mdpi.com/2076-3417/15/6/2902<i>Lactobacillus</i>probiotic strainsprobiotic propertiesprobioticsbiological activities
spellingShingle Yulian Tumbarski
Ivelina Peykova-Shapkova
Mihaela Ivanova
Remzi Cholakov
Agata Dutkiewicz
Krzysztof Grzymajło
Characterization and Selection of <i>Lactobacillus</i> Strains with Potential Probiotic Applications
Applied Sciences
<i>Lactobacillus</i>
probiotic strains
probiotic properties
probiotics
biological activities
title Characterization and Selection of <i>Lactobacillus</i> Strains with Potential Probiotic Applications
title_full Characterization and Selection of <i>Lactobacillus</i> Strains with Potential Probiotic Applications
title_fullStr Characterization and Selection of <i>Lactobacillus</i> Strains with Potential Probiotic Applications
title_full_unstemmed Characterization and Selection of <i>Lactobacillus</i> Strains with Potential Probiotic Applications
title_short Characterization and Selection of <i>Lactobacillus</i> Strains with Potential Probiotic Applications
title_sort characterization and selection of i lactobacillus i strains with potential probiotic applications
topic <i>Lactobacillus</i>
probiotic strains
probiotic properties
probiotics
biological activities
url https://www.mdpi.com/2076-3417/15/6/2902
work_keys_str_mv AT yuliantumbarski characterizationandselectionofilactobacillusistrainswithpotentialprobioticapplications
AT ivelinapeykovashapkova characterizationandselectionofilactobacillusistrainswithpotentialprobioticapplications
AT mihaelaivanova characterizationandselectionofilactobacillusistrainswithpotentialprobioticapplications
AT remzicholakov characterizationandselectionofilactobacillusistrainswithpotentialprobioticapplications
AT agatadutkiewicz characterizationandselectionofilactobacillusistrainswithpotentialprobioticapplications
AT krzysztofgrzymajło characterizationandselectionofilactobacillusistrainswithpotentialprobioticapplications