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In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (Capra hircus)

SIMPLE SUMMARY: This is the first report of highly safe and potential probiotic strains, Lacticaseibacillus rhamnosus YM2-1 and YM2-3, isolated from the raw milk of dairy Japanese-Saanen goats. The antipathogenic activities of the two strains were identified based on their probiotic features. Althou...

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Autores principales: Tanaka, Yukimune, Aryantini, Ni Putu Desy, Yamasaki, Eiki, Saito, Makoto, Tsukigase, Yui, Nakatsuka, Hirotaka, Urashima, Tadasu, Horiuchi, Risa, Fukuda, Kenji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9817676/
https://www.ncbi.nlm.nih.gov/pubmed/36611617
http://dx.doi.org/10.3390/ani13010007
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author Tanaka, Yukimune
Aryantini, Ni Putu Desy
Yamasaki, Eiki
Saito, Makoto
Tsukigase, Yui
Nakatsuka, Hirotaka
Urashima, Tadasu
Horiuchi, Risa
Fukuda, Kenji
author_facet Tanaka, Yukimune
Aryantini, Ni Putu Desy
Yamasaki, Eiki
Saito, Makoto
Tsukigase, Yui
Nakatsuka, Hirotaka
Urashima, Tadasu
Horiuchi, Risa
Fukuda, Kenji
author_sort Tanaka, Yukimune
collection PubMed
description SIMPLE SUMMARY: This is the first report of highly safe and potential probiotic strains, Lacticaseibacillus rhamnosus YM2-1 and YM2-3, isolated from the raw milk of dairy Japanese-Saanen goats. The antipathogenic activities of the two strains were identified based on their probiotic features. Although L. rhamnosus is on the list of qualified presumption of safety (QPS)-recommended biological agents assessed by the European Food Safety Authority (EFSA), additional in vitro examinations performed in this study ensure the safety of both strains, considering the risk of possible strain-dependent unfavorable characteristics. ABSTRACT: Two novel probiotic strains of lactic acid bacteria were successfully isolated from the raw milk of dairy Japanese-Saanen goats. Selection criteria for positive candidates were grown on de Man–Rogosa–Sharpe or M17 selective medium at 30, 35, or 42 °C anaerobically, and characterized based on Gram reaction, catalase test, and tolerance to low pH and bile salts. Among the 101 isolated positive candidates, two strains, YM2-1 and YM2-3, were selected and identified as Lacticaseibacillus rhamnosus using 16S rDNA sequence similarity. Culture supernatants of the two strains exhibited antipathogenic activity against Salmonella enterica subsp. enterica serovar. Typhimurium, Shigella sonnei, methicillin-resistant Staphylococcus aureus, methicillin-sensitive Staphylococcus aureus, Listeria monocytogenes, and Escherichia coli O157. The antipathogenic activities were retained to some extent after neutralization, indicating the presence of antipathogenic substances other than organic acids in the culture supernatants. The two strains were sensitive with coincidental minimum inhibition concentrations (indicated in the parentheses hereafter) to ampicillin (0.25 μg/mL), chloramphenicol (4 μg/mL), gentamycin (4 μg/mL), kanamycin (64 μg/mL), streptomycin (16 μg/mL), and tetracycline (4 μg/mL). Furthermore, the two strains were resistant to clindamycin (16 μg/mL) and erythromycin (4 μg/mL). In addition, both YM2-1 and YM2-3 strains showed less unfavorable activities, including bile acid bioconversion, carcinogenic-related enzymes, mucin degradation, plasminogen activation, and hemolysis, than the detection limits of in vitro evaluation methods used in this study. In summary, L. rhamnosus YM2-1 and YM2-3 are highly safe and promising probiotic strains applicable in the dairy industry, and were first isolated from the raw milk of Japanese-Saanen goats.
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spelling pubmed-98176762023-01-07 In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (Capra hircus) Tanaka, Yukimune Aryantini, Ni Putu Desy Yamasaki, Eiki Saito, Makoto Tsukigase, Yui Nakatsuka, Hirotaka Urashima, Tadasu Horiuchi, Risa Fukuda, Kenji Animals (Basel) Article SIMPLE SUMMARY: This is the first report of highly safe and potential probiotic strains, Lacticaseibacillus rhamnosus YM2-1 and YM2-3, isolated from the raw milk of dairy Japanese-Saanen goats. The antipathogenic activities of the two strains were identified based on their probiotic features. Although L. rhamnosus is on the list of qualified presumption of safety (QPS)-recommended biological agents assessed by the European Food Safety Authority (EFSA), additional in vitro examinations performed in this study ensure the safety of both strains, considering the risk of possible strain-dependent unfavorable characteristics. ABSTRACT: Two novel probiotic strains of lactic acid bacteria were successfully isolated from the raw milk of dairy Japanese-Saanen goats. Selection criteria for positive candidates were grown on de Man–Rogosa–Sharpe or M17 selective medium at 30, 35, or 42 °C anaerobically, and characterized based on Gram reaction, catalase test, and tolerance to low pH and bile salts. Among the 101 isolated positive candidates, two strains, YM2-1 and YM2-3, were selected and identified as Lacticaseibacillus rhamnosus using 16S rDNA sequence similarity. Culture supernatants of the two strains exhibited antipathogenic activity against Salmonella enterica subsp. enterica serovar. Typhimurium, Shigella sonnei, methicillin-resistant Staphylococcus aureus, methicillin-sensitive Staphylococcus aureus, Listeria monocytogenes, and Escherichia coli O157. The antipathogenic activities were retained to some extent after neutralization, indicating the presence of antipathogenic substances other than organic acids in the culture supernatants. The two strains were sensitive with coincidental minimum inhibition concentrations (indicated in the parentheses hereafter) to ampicillin (0.25 μg/mL), chloramphenicol (4 μg/mL), gentamycin (4 μg/mL), kanamycin (64 μg/mL), streptomycin (16 μg/mL), and tetracycline (4 μg/mL). Furthermore, the two strains were resistant to clindamycin (16 μg/mL) and erythromycin (4 μg/mL). In addition, both YM2-1 and YM2-3 strains showed less unfavorable activities, including bile acid bioconversion, carcinogenic-related enzymes, mucin degradation, plasminogen activation, and hemolysis, than the detection limits of in vitro evaluation methods used in this study. In summary, L. rhamnosus YM2-1 and YM2-3 are highly safe and promising probiotic strains applicable in the dairy industry, and were first isolated from the raw milk of Japanese-Saanen goats. MDPI 2022-12-20 /pmc/articles/PMC9817676/ /pubmed/36611617 http://dx.doi.org/10.3390/ani13010007 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Tanaka, Yukimune
Aryantini, Ni Putu Desy
Yamasaki, Eiki
Saito, Makoto
Tsukigase, Yui
Nakatsuka, Hirotaka
Urashima, Tadasu
Horiuchi, Risa
Fukuda, Kenji
In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (Capra hircus)
title In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (Capra hircus)
title_full In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (Capra hircus)
title_fullStr In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (Capra hircus)
title_full_unstemmed In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (Capra hircus)
title_short In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (Capra hircus)
title_sort in vitro probiotic characterization and safety assessment of lactic acid bacteria isolated from raw milk of japanese-saanen goat (capra hircus)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9817676/
https://www.ncbi.nlm.nih.gov/pubmed/36611617
http://dx.doi.org/10.3390/ani13010007
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