{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T05:19:01Z","timestamp":1788153541562,"version":"build-2803163510"},"reference-count":33,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"\n Introduction<\/jats:title>\n \n Biofilm formation by\n Streptococcus agalactiae<\/jats:italic>\n (\n S. agalactiae<\/jats:italic>\n ) is a major driver of chronic bovine mastitis that resists conventional antibiotic treatment, yet targeted strategies that disarm its biofilm machinery remain limited.\n Sophora flavescens (S. flavescens)<\/jats:italic>\n is a medicinal plant rich in prenylated flavonoids with antimicrobial properties, but the antibiofilm mechanisms of these compounds against\n S. agalactiae<\/jats:italic>\n remain unexplored.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n The antibacterial activity of 15 prenylated flavonoids from\n S. flavescens<\/jats:italic>\n was evaluated against major mastitis-associated bacteria, and minimum inhibitory concentrations (MIC\u2089\u2080) were determined. Biofilm inhibition was assessed by crystal violet and acridine orange staining. Molecular docking, quantitative real-time PCR (qPCR), untargeted metabolomics, and phenotypic validation assays were integrated to elucidate the antibiofilm mechanisms. A mouse mastitis model was employed to evaluate\n in vivo<\/jats:italic>\n efficacy, and cytotoxicity and acute toxicity assays were performed to assess safety.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Kurarinone and sophoraflavanone G exhibited potent antibacterial activity against\n S. agalactiae<\/jats:italic>\n ,\n Streptococcus dysgalactiae<\/jats:italic>\n , and\n Staphylococcus aureus<\/jats:italic>\n , with MIC\u2089\u2080 values of 3.12\u202f\u03bcg\/mL against\n S. agalactiae<\/jats:italic>\n , superior to norfloxacin (6.25\u202f\u03bcg\/mL). Mechanistically, these compounds acted through two complementary pathways: (1) strong binding affinity to the adhesion-related proteins FbsA, FbsB, FbsC, and Lmb in molecular docking analysis, with transcriptional downregulation of their encoding genes confirmed by qPCR, and (2) perturbation of the ATP-binding cassette (ABC) transporter pathway, as suggested by untargeted metabolomics, leading to markedly reduced secretion of extracellular polysaccharides (by up to 60%), capsular polysaccharides, and extracellular DNA. Notably, this dual antibiofilm activity occurred at sub-MIC concentrations, indicating an antibiofilm mechanism that operates independently of direct bacterial killing and is distinct from the previously reported membrane-disruptive effects of these compounds. In a mouse mastitis model, intragastric administration of both compounds significantly ameliorated mammary tissue histopathology, reduced pro-inflammatory cytokines (TNF-\n \u03b1<\/jats:italic>\n , IL-1\u03b2, IL-6, IL-8), and decreased myeloperoxidase activity, with favorable safety profiles (LD\u2085\u2080\u202f&gt;\u202f2000\u202fmg\/kg).\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n These findings suggest that adhesion proteins and the ABC transporter pathway represent potential dual targets against\n S. agalactiae<\/jats:italic>\n biofilms, and position kurarinone and sophoraflavanone G as promising leads for antibiofilm drug development targeting biofilm-associated bovine mastitis.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1905109","type":"journal-article","created":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T04:29:44Z","timestamp":1788150584000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Kurarinone and sophoraflavanone G from Sophora flavescens suppress Streptococcus agalactiae biofilm via dual antibiofilm mechanisms involving adhesion protein binding and ABC transporter pathway perturbation"],"prefix":"10.3389","volume":"17","author":[{"given":"Jihao","family":"Shi","sequence":"first","affiliation":[{"name":"College of Pharmacy and Food, Southwest Minzu University","place":["Chengdu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yun","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Food, Southwest Minzu University","place":["Chengdu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junxia","family":"An","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Food, Southwest Minzu University","place":["Chengdu, China"]},{"name":"Academy of Agricultural and Forestry Sciences, Qinghai University","place":["Xining, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linglin","family":"Li","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Food, Southwest Minzu University","place":["Chengdu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anping","family":"Li","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Food, Southwest Minzu University","place":["Chengdu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"17576","DOI":"10.1021\/acs.jafc.5c01595","article-title":"Antibacterial activities of prenylated flavonoids from Sophora flavescens against Xanthomonas oryzae pv. 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