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Their use in drug development is often inspired by their traditional or historical medical use. For many decades, this discovery pipeline has focused on identifying a single molecule responsible for much of the biological activity of a \u201craw\u201d natural product preparation (e.g., a whole-plant extract) and scoping this molecule for clinical potential. However, it is increasingly realized that historical\/traditional remedies with significant biological activity may owe this activity to the combined action of multiple molecules. Concomitantly, microbiologists increasingly argue that effectively fighting antimicrobial-resistant infections will rely on combination therapies that combine multiple antimicrobials and\/or adjuvant molecules. We previously reconstructed a complex historical remedy, Bald\u2019s eyesalve. Our reconstruction of this remedy had strong antibiofilm activity, which relied on the presence of multiple ingredients. Here, we report that Bald\u2019s eyesalve has multiple antibacterial effects on exemplar gram-positive (\n                      <jats:italic toggle=\"yes\">Staphylococcus aureus<\/jats:italic>\n                      ) and gram-negative (\n                      <jats:italic toggle=\"yes\">Acinetobacter baumannii<\/jats:italic>\n                      ) pathogens. Bald\u2019s eyesalve disrupts bacterial membrane integrity; inhibits the expression of genes associated with bacterial adhesins, virulence factors, and efflux pumps in both\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">A. baumannii<\/jats:italic>\n                      ; inhibits quorum sensing in\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      ; and causes the downregulation of genes involved in\n                      <jats:italic toggle=\"yes\">de novo<\/jats:italic>\n                      nucleotide biosynthesis in\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      . Finally, we show that this multifaceted mechanism of action makes it difficult for\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">A. baumannii<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">Pseudomonas aeruginosa<\/jats:italic>\n                      to evolve resistance against Bald\u2019s eyesalve. Bald\u2019s eyesalve could be used to identify a defined cocktail of natural products suitable for preclinical testing as a multi-target antibacterial preparation to which resistance may arise more slowly than current single-molecule antibiotics.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>The increasing mortality and economic cost of antimicrobial resistance (AMR) have made it one of the biggest threats to global health. Available antibiotics are in short supply due to the slow progress in the discovery of new antibiotics. Hence, there is a need for alternative treatment options against difficult-to-treat infections. We have identified a natural product cocktail based on a historical remedy with broad-spectrum antibacterial activity and a multifaceted mechanism of action. We have shown that there is slower resistance evolution to this cocktail compared to mainline antibiotics, and it could be used as the foundation of an alternative treatment to antibiotics in clinical settings.<\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/msphere.00494-26","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T13:00:22Z","timestamp":1788440422000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Uncovering the mechanism of action of a historical antimicrobial against\n                    <i>S. aureus<\/i>\n                    and\n                    <i>A. baumannii<\/i>"],"prefix":"10.1128","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8905-9927","authenticated-orcid":true,"given":"Oluwatosin 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