16S-Amplified Ribosomal DNA Restriction Analysis Assay for Discriminating Potentially Probiotic Lactobacillus Species Isolated from Traditional Dairy Products
Background: Traditional dairy products are the main sources for probiotic bacteria. This study aimed to isolate and characterize the potentially probiotic Lactobacillus strains isolated from traditional dairy products in Iran. Methods: Microbial population of each dairy product was enriched and scre...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Kerman University of Medical Sciences
2018-03-01
|
| Series: | Journal of Kerman University of Medical Sciences |
| Subjects: | |
| Online Access: | https://jkmu.kmu.ac.ir/article_66887_f67310c67edab8f9704ae66867b03473.pdf |
| _version_ | 1797790671211331584 |
|---|---|
| author | Soudeh Ghafouri-Fard Mohammad Amin Hejazi Davoud Afshar Abolfazl Barzegari Solat Eslami |
| author_facet | Soudeh Ghafouri-Fard Mohammad Amin Hejazi Davoud Afshar Abolfazl Barzegari Solat Eslami |
| author_sort | Soudeh Ghafouri-Fard |
| collection | DOAJ |
| description | Background: Traditional dairy products are the main sources for probiotic bacteria. This study aimed to isolate and characterize the potentially probiotic Lactobacillus strains isolated from traditional dairy products in Iran. Methods: Microbial population of each dairy product was enriched and screened for acid- and bile- resistant strains. The isolates were primarily characterized by conventional phenotypic and biochemical tests. The species were finally identified using restriction analysis and 16S rDNA sequencing. In addition, the antibacterial activity of the four most common species was evaluated against seven gastrointestinal pathogens. Results: From a total of nine yogurt and cheese samples, 17 acid-resistant Lactobacillus strains were isolated (survival>50% in PBS buffer, pH 2.5 for 3 h). Eight of them also showed high tolerance to bile. Also, the superiority of molecular techniques over phenotypic tests for effective identification of species was observed. Biochemical tests were useful in combination with 16S-amplified ribosomal DNA restriction analysis (ARDRA) technique to discriminate the isolates at species level. Conclusion: 16S–ARDRA in combination with biochemical tests is suggested as a potentially useful method for accurate identification of Lactobacillus species where the 16S-rDNA sequencing could not discriminate between two closely related species. Considering the acid and bile resistance capacity of isolated strains, traditional dairy products may be used as important sources of probiotic strains. |
| first_indexed | 2024-03-13T02:07:52Z |
| format | Article |
| id | doaj.art-210688653e2b4a41878ebe48285c1961 |
| institution | Directory Open Access Journal |
| issn | 2008-2843 |
| language | English |
| last_indexed | 2024-03-13T02:07:52Z |
| publishDate | 2018-03-01 |
| publisher | Kerman University of Medical Sciences |
| record_format | Article |
| series | Journal of Kerman University of Medical Sciences |
| spelling | doaj.art-210688653e2b4a41878ebe48285c19612023-07-01T05:17:19ZengKerman University of Medical SciencesJournal of Kerman University of Medical Sciences2008-28432018-03-012521751866688716S-Amplified Ribosomal DNA Restriction Analysis Assay for Discriminating Potentially Probiotic Lactobacillus Species Isolated from Traditional Dairy ProductsSoudeh Ghafouri-Fard0Mohammad Amin Hejazi1Davoud Afshar2Abolfazl Barzegari3Solat Eslami4Associate Professor of Medical Genetics, Department of Medical Genetics, Shahid Beheshti University of Medical Sciences, Tehran, IranAssociate Professor of Food Biotechnology, Department of Food Biotechnology, Branch for Northwest & West region, Agricultural Biotechnology Research Institute of Iran, Agricultural Research, Education and Extension Organization (AREEO), Tabriz, IranAssistant Professor of Medical Bacteriology, Department of Microbiology and Virology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, IranPh.D. Candidate of Medical Biotechnology, Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz, IranAssistant Professor of Medical Biotechnology, Dietary Supplements and Probiotic Research Center, Alborz University of Medical Sciences, Karaj, IranBackground: Traditional dairy products are the main sources for probiotic bacteria. This study aimed to isolate and characterize the potentially probiotic Lactobacillus strains isolated from traditional dairy products in Iran. Methods: Microbial population of each dairy product was enriched and screened for acid- and bile- resistant strains. The isolates were primarily characterized by conventional phenotypic and biochemical tests. The species were finally identified using restriction analysis and 16S rDNA sequencing. In addition, the antibacterial activity of the four most common species was evaluated against seven gastrointestinal pathogens. Results: From a total of nine yogurt and cheese samples, 17 acid-resistant Lactobacillus strains were isolated (survival>50% in PBS buffer, pH 2.5 for 3 h). Eight of them also showed high tolerance to bile. Also, the superiority of molecular techniques over phenotypic tests for effective identification of species was observed. Biochemical tests were useful in combination with 16S-amplified ribosomal DNA restriction analysis (ARDRA) technique to discriminate the isolates at species level. Conclusion: 16S–ARDRA in combination with biochemical tests is suggested as a potentially useful method for accurate identification of Lactobacillus species where the 16S-rDNA sequencing could not discriminate between two closely related species. Considering the acid and bile resistance capacity of isolated strains, traditional dairy products may be used as important sources of probiotic strains.https://jkmu.kmu.ac.ir/article_66887_f67310c67edab8f9704ae66867b03473.pdfdairy productsprobioticslactobacillusmolecular techniques16s-ardra |
| spellingShingle | Soudeh Ghafouri-Fard Mohammad Amin Hejazi Davoud Afshar Abolfazl Barzegari Solat Eslami 16S-Amplified Ribosomal DNA Restriction Analysis Assay for Discriminating Potentially Probiotic Lactobacillus Species Isolated from Traditional Dairy Products Journal of Kerman University of Medical Sciences dairy products probiotics lactobacillus molecular techniques 16s-ardra |
| title | 16S-Amplified Ribosomal DNA Restriction Analysis Assay for Discriminating Potentially Probiotic Lactobacillus Species Isolated from Traditional Dairy Products |
| title_full | 16S-Amplified Ribosomal DNA Restriction Analysis Assay for Discriminating Potentially Probiotic Lactobacillus Species Isolated from Traditional Dairy Products |
| title_fullStr | 16S-Amplified Ribosomal DNA Restriction Analysis Assay for Discriminating Potentially Probiotic Lactobacillus Species Isolated from Traditional Dairy Products |
| title_full_unstemmed | 16S-Amplified Ribosomal DNA Restriction Analysis Assay for Discriminating Potentially Probiotic Lactobacillus Species Isolated from Traditional Dairy Products |
| title_short | 16S-Amplified Ribosomal DNA Restriction Analysis Assay for Discriminating Potentially Probiotic Lactobacillus Species Isolated from Traditional Dairy Products |
| title_sort | 16s amplified ribosomal dna restriction analysis assay for discriminating potentially probiotic lactobacillus species isolated from traditional dairy products |
| topic | dairy products probiotics lactobacillus molecular techniques 16s-ardra |
| url | https://jkmu.kmu.ac.ir/article_66887_f67310c67edab8f9704ae66867b03473.pdf |
| work_keys_str_mv | AT soudehghafourifard 16samplifiedribosomaldnarestrictionanalysisassayfordiscriminatingpotentiallyprobioticlactobacillusspeciesisolatedfromtraditionaldairyproducts AT mohammadaminhejazi 16samplifiedribosomaldnarestrictionanalysisassayfordiscriminatingpotentiallyprobioticlactobacillusspeciesisolatedfromtraditionaldairyproducts AT davoudafshar 16samplifiedribosomaldnarestrictionanalysisassayfordiscriminatingpotentiallyprobioticlactobacillusspeciesisolatedfromtraditionaldairyproducts AT abolfazlbarzegari 16samplifiedribosomaldnarestrictionanalysisassayfordiscriminatingpotentiallyprobioticlactobacillusspeciesisolatedfromtraditionaldairyproducts AT solateslami 16samplifiedribosomaldnarestrictionanalysisassayfordiscriminatingpotentiallyprobioticlactobacillusspeciesisolatedfromtraditionaldairyproducts |