In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (<i>Capra hircus</i>)
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 re...
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MDPI AG
2022-12-01
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| author | Yukimune Tanaka Ni Putu Desy Aryantini Eiki Yamasaki Makoto Saito Yui Tsukigase Hirotaka Nakatsuka Tadasu Urashima Risa Horiuchi Kenji Fukuda |
| author_facet | Yukimune Tanaka Ni Putu Desy Aryantini Eiki Yamasaki Makoto Saito Yui Tsukigase Hirotaka Nakatsuka Tadasu Urashima Risa Horiuchi Kenji Fukuda |
| author_sort | Yukimune Tanaka |
| collection | DOAJ |
| description | 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 <i>Lacticaseibacillus rhamnosus</i> using 16S rDNA sequence similarity. Culture supernatants of the two strains exhibited antipathogenic activity against <i>Salmonella enterica</i> subsp. <i>enterica</i> serovar. Typhimurium, <i>Shigella sonnei</i>, methicillin-resistant <i>Staphylococcus aureus</i>, methicillin-sensitive <i>Staphylococcus aureus</i>, <i>Listeria monocytogenes</i>, and <i>Escherichia coli</i> 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, <i>L. rhamnosus</i> 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. |
| first_indexed | 2024-03-11T10:12:32Z |
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| institution | Directory Open Access Journal |
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| last_indexed | 2024-03-11T10:12:32Z |
| publishDate | 2022-12-01 |
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| spelling | doaj.art-2b8b98a89d3c45d7986407a36e58b8d22023-11-16T14:17:54ZengMDPI AGAnimals2076-26152022-12-01131710.3390/ani13010007In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (<i>Capra hircus</i>)Yukimune Tanaka0Ni Putu Desy Aryantini1Eiki Yamasaki2Makoto Saito3Yui Tsukigase4Hirotaka Nakatsuka5Tadasu Urashima6Risa Horiuchi7Kenji Fukuda8Department of Life and Food Sciences, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada-cho, Obihiro, Hokkaido 080-8555, JapanDepartment of Life and Food Sciences, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada-cho, Obihiro, Hokkaido 080-8555, JapanDepartment of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada-cho, Obihiro, Hokkaido 080-8555, JapanKisara Farm Cheese Factory, Haobiminami 10, Shimizu-cho, Kamikawa-gun, Hokkaido 089-0356, JapanDepartment of Life and Food Sciences, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada-cho, Obihiro, Hokkaido 080-8555, JapanDepartment of Life and Food Sciences, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada-cho, Obihiro, Hokkaido 080-8555, JapanDepartment of Life and Food Sciences, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada-cho, Obihiro, Hokkaido 080-8555, JapanDepartment of Life and Food Sciences, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada-cho, Obihiro, Hokkaido 080-8555, JapanDepartment of Life and Food Sciences, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada-cho, Obihiro, Hokkaido 080-8555, JapanTwo 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 <i>Lacticaseibacillus rhamnosus</i> using 16S rDNA sequence similarity. Culture supernatants of the two strains exhibited antipathogenic activity against <i>Salmonella enterica</i> subsp. <i>enterica</i> serovar. Typhimurium, <i>Shigella sonnei</i>, methicillin-resistant <i>Staphylococcus aureus</i>, methicillin-sensitive <i>Staphylococcus aureus</i>, <i>Listeria monocytogenes</i>, and <i>Escherichia coli</i> 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, <i>L. rhamnosus</i> 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.https://www.mdpi.com/2076-2615/13/1/7antipathogenic activityantimicrobial resistancecheese starter<i>Lacticaseibacillus rhamnosus</i> |
| spellingShingle | Yukimune Tanaka Ni Putu Desy Aryantini Eiki Yamasaki Makoto Saito Yui Tsukigase Hirotaka Nakatsuka Tadasu Urashima Risa Horiuchi Kenji Fukuda In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (<i>Capra hircus</i>) Animals antipathogenic activity antimicrobial resistance cheese starter <i>Lacticaseibacillus rhamnosus</i> |
| title | In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (<i>Capra hircus</i>) |
| title_full | In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (<i>Capra hircus</i>) |
| title_fullStr | In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (<i>Capra hircus</i>) |
| title_full_unstemmed | In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (<i>Capra hircus</i>) |
| title_short | In Vitro Probiotic Characterization and Safety Assessment of Lactic Acid Bacteria Isolated from Raw Milk of Japanese-Saanen Goat (<i>Capra hircus</i>) |
| title_sort | in vitro probiotic characterization and safety assessment of lactic acid bacteria isolated from raw milk of japanese saanen goat i capra hircus i |
| topic | antipathogenic activity antimicrobial resistance cheese starter <i>Lacticaseibacillus rhamnosus</i> |
| url | https://www.mdpi.com/2076-2615/13/1/7 |
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