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Compared with planktonic cells, biofilm-associated microorganisms can exhibit up to 10- to 1000-fold increased tolerance to antimicrobial agents, contributing to the persistence of biofilm-associated infections (BAIs). These infections remain difficult to eradicate due to reduced penetration, altered metabolic states, and the presence of dormant or persister cells. Anti-biofilm strategies can be broadly classified into physical approaches (e.g., ultrasound, mechanical stress, and light-based approaches) that target biofilm structure; chemical and enzymatic methods (e.g., EPS-degrading enzymes) that destabilize the matrix; and biological and molecular strategies (e.g., quorum-sensing (QS) inhibitors, anti-virulence agents, bacteriophages, phage-derived antimicrobial molecules, antimicrobial peptides, and natural bioactive compounds) that modulate biofilm development and integrity by targeting regulatory pathways and matrix stability through distinct mechanisms of action. Natural compounds, including lactoferrin, lactoferrin-derived peptides, and probiotic and postbiotic fractions of lactic acid bacteria (LAB), as well as plant-derived metabolites, have shown promising anti-biofilm effects, with efficacy often enhanced through complementary or potentially synergistic interactions. However, despite these advancements, clinical translation remains limited. For example, BAIs account for approximately 80% of chronic infections, with high recurrence rates and therapeutic failure reported in device-associated infections and chronic wounds. These limitations highlight the need for clinically translatable, multimodal approaches that integrate structural biofilm disruption, antimicrobial targeting, and host response modulation to design more effective and sustainable anti-biofilm strategies.<\/jats:p>","DOI":"10.3390\/biom16060887","type":"journal-article","created":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T00:49:42Z","timestamp":1781743782000},"page":"887","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Innovative Strategies to Abolish Microbial Persistence in Biofilm Fortresses"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-9625-8922","authenticated-orcid":false,"given":"Diana-Antonia","family":"Costea","sequence":"first","affiliation":[{"name":"Faculty of Biology, University of Bucharest, Splaiul Independentei 91\u201395, R-050095 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-3455-1945","authenticated-orcid":false,"given":"Valentina-Alexandra","family":"Badaluta","sequence":"additional","affiliation":[{"name":"Faculty of Biology, University of Bucharest, Splaiul Independentei 91\u201395, R-050095 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-2738-6926","authenticated-orcid":false,"given":"Ioana","family":"Zachia-Zlatea","sequence":"additional","affiliation":[{"name":"Faculty of Biology, University of Bucharest, Splaiul Independentei 91\u201395, R-050095 Bucharest, Romania"},{"name":"Doctoral School of Urban Planning, \u201cIon Mincu\u201d University of Architecture and Urbanism, Academiei Street 18\u201320, R-010014 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alina-Maria","family":"Holban","sequence":"additional","affiliation":[{"name":"Faculty of Biology, University of Bucharest, Splaiul Independentei 91\u201395, R-050095 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6593-255X","authenticated-orcid":false,"given":"Lia-Mara","family":"Ditu","sequence":"additional","affiliation":[{"name":"Faculty of Biology, University of Bucharest, Splaiul Independentei 91\u201395, R-050095 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veronica","family":"Lazar","sequence":"additional","affiliation":[{"name":"Faculty of Biology, University of Bucharest, Splaiul Independentei 91\u201395, R-050095 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1038\/nrmicro.2016.94","article-title":"Biofilms: An Emergent Form of Bacterial Life","volume":"14","author":"Flemming","year":"2016","journal-title":"Nat. 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