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Probiotics could be a promising alternative for fighting antibiotic resistance, displaying beneficial effects to the host by combating diseases, improving growth, and stimulating the host immune responses against infection. This study was conducted to evaluate the probiotic, antibacterial, and antibiofilm potential of <jats:italic>Streptomyces levis<\/jats:italic> strain HFM-2 isolated from the healthy human gut.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>In vitro antibacterial activity in the cell-free supernatant of <jats:italic>S. levis<\/jats:italic> strain HFM-2 was evaluated against different pathogens viz. <jats:italic>K. pneumoniae<\/jats:italic> sub sp. <jats:italic>pneumoniae, S. aureus, B. subtilis<\/jats:italic>, VRE, <jats:italic>S. typhi, S. epidermidis<\/jats:italic>, MRSA, <jats:italic>V. cholerae, M. smegmatis, E. coli, P. aeruginosa<\/jats:italic> and <jats:italic>E. aerogenes<\/jats:italic>. Further, the ethyl acetate extract from <jats:italic>S. levis<\/jats:italic> strain HFM-2 showed strong biofilm inhibition against <jats:italic>S. typhi, K. pneumoniae<\/jats:italic> sub sp. <jats:italic>pneumoniae, P. aeruginosa<\/jats:italic> and <jats:italic>E. coli.<\/jats:italic> Fluorescence microscopy was used to detect biofilm inhibition properties. MIC and MBC values of EtOAc extract were determined at 500 and 1000\u00a0\u00b5g\/mL, respectively. Further, strain HFM-2 showed high tolerance in gastric juice, pancreatin, bile, and at low pH. It exhibited efficient adhesion properties, displaying auto-aggregation (97.0%), hydrophobicity (95.71%, 88.96%, and 81.15% for ethyl acetate, chloroform and xylene, respectively), and showed 89.75%, 86.53%, 83.06% and 76.13% co-aggregation with <jats:italic>S. typhi<\/jats:italic>, MRSA, <jats:italic>S. pyogenes<\/jats:italic> and <jats:italic>E. coli<\/jats:italic>, respectively after 60\u00a0min of incubation. The <jats:italic>S. levis<\/jats:italic> strain HFM-2 was susceptible to different antibiotics such as tetracycline, streptomycin, kanamycin, ciprofloxacin, erythromycin, linezolid, meropenem, amikacin, gentamycin, clindamycin, moxifloxacin and vancomycin, but resistant to ampicillin and penicillin G.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusion<\/jats:title>\n                <jats:p>The study shows that <jats:italic>S. levis<\/jats:italic> strain HFM-2 has significant probiotic properties such as good viability in bile, gastric juice, pancreatin environment, and at low pH; proficient adhesion properties, and antibiotic susceptibility. Further, the EtOAc extract of <jats:italic>Streptomyces levis<\/jats:italic> strain HFM-2 has a potent antibiofilm and antibacterial activity against antibacterial-resistant clinical pathogens.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s12866-024-03353-x","type":"journal-article","created":{"date-parts":[[2024,6,11]],"date-time":"2024-06-11T11:14:21Z","timestamp":1718104461000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Probiotic potential of Streptomyces levis strain HFM-2 isolated from human gut and its antibiofilm properties against pathogenic bacteria"],"prefix":"10.1186","volume":"24","author":[{"given":"Jaya","family":"Verma","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sapna","family":"Devi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anmol","family":"Narang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sukhraj","family":"Kaur","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rajesh Kumari","family":"Manhas","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,6,11]]},"reference":[{"key":"3353_CR1","doi-asserted-by":"publisher","first-page":"2928doi","DOI":"10.3389\/fmicb.2018.02928","volume":"9","author":"E Peterson","year":"2018","unstructured":"Peterson E, Kaur P. 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