{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T21:50:29Z","timestamp":1793397029728,"version":"4.5.0"},"reference-count":73,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T00:00:00Z","timestamp":1779667200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Royal Higher Institute for Defence","award":["HFM-21-04"],"award-info":[{"award-number":["HFM-21-04"]}]},{"award":["HFM-21-04"],"award-info":[{"award-number":["HFM-21-04"]}],"id":[{"id":"https:\/\/ror.org\/00kxpat18","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Background\/Objectives: Biofilms are a form of defense that enables bacteria to withstand antibiotic pressure and demonstrate antibiotic resistance. It is crucial to develop anti-biofilm strategies in order to combat chronic and persistent multidrug-resistant (MDR) infections. Methods: In this study, we developed 3D biofilms of single-, dual-, and poly-species MDR ESKAPE components, including the pathogens P. aeruginosa S. aureus and K. pneumoniae, in CF Mu3Gel. We evaluated the efficacy of using a phage, a di-hetero phage cocktail or a poly-hetero phage cocktail in combination with ciprofloxacin to eliminate mature biofilm biomass after 72 h or one week in a single treatment. Results: The phage components mostly exhibited synergistic behavior when combined with ciprofloxacin and with each other in di- and poly-hetero-cocktails. The reduction in 72-h dual- and poly-species biofilms was one log higher than that of one-week biofilms treated with the phage\u2013antibiotic combination. The greatest reductions were observed in the 72-h single-species biofilm with combination therapy, at 1.4\u20133.0 log. Reductions of 2.16 and 1.6 log were observed in the dual-species P. aeruginosa and S. aureus biofilm and the poly-species biofilm, respectively. Conclusions: This study examined how a single application of phages or phage cocktails, either alone or in combination with ciprofloxacin, impacted established biofilm models, and how this affected the proportion of microcolonies of different species within each model. These insights will facilitate the development of strategies for multiple follow-up treatments, as well as the reordering of phages, phage cocktails, and combinations with antibiotics, to improve outcomes. The 3D biofilm models developed here could be used to screen phages or phage cocktails either on their own or alongside other therapies. This would facilitate the application of in vitro findings to real physiological settings.<\/jats:p>","DOI":"10.3390\/antibiotics15060537","type":"journal-article","created":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T14:10:22Z","timestamp":1779718222000},"page":"537","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Single Treatment of Mature 3D Single-, Dual- and Poly-Species Biofilms Using a Combination Therapy of Phage or Phage-Hetero-Cocktails and Ciprofloxacin"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5814-3563","authenticated-orcid":false,"given":"Tea","family":"Glonti","sequence":"first","affiliation":[{"name":"Laboratory for Molecular and Cellular Technology, Queen Astrid Military Hospital, Bruynstraat 1, 1120 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6014-1580","authenticated-orcid":false,"given":"Merve K\u00fcbra","family":"Aktan","sequence":"additional","affiliation":[{"name":"Biomaterials and Tissue Engineering Research Group, Department of Materials Engineering (MTM), KU Leuven, Kasteelpark Arenberg 44, 3001 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christel","family":"Cochez","sequence":"additional","affiliation":[{"name":"Laboratory for Molecular and Cellular Technology, Queen Astrid Military Hospital, Bruynstraat 1, 1120 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7773-8199","authenticated-orcid":false,"given":"Naiera","family":"Zayed","sequence":"additional","affiliation":[{"name":"Periodontology & Oral Microbiology, Oral Health Sciences, KU Leuven, Kapucijnenvoer 7, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4890-0177","authenticated-orcid":false,"given":"Annabel","family":"Braem","sequence":"additional","affiliation":[{"name":"Biomaterials and Tissue Engineering Research Group, Department of Materials Engineering (MTM), KU Leuven, Kasteelpark Arenberg 44, 3001 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wim","family":"Teughels","sequence":"additional","affiliation":[{"name":"Periodontology & Oral Microbiology, Oral Health Sciences, KU Leuven, Kapucijnenvoer 7, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6863-5219","authenticated-orcid":false,"given":"Jean-Paul","family":"Pirnay","sequence":"additional","affiliation":[{"name":"Laboratory for Molecular and Cellular Technology, Queen Astrid Military Hospital, Bruynstraat 1, 1120 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"270","DOI":"10.3390\/antibiotics3030270","article-title":"Bacteriophages and Biofilms","volume":"3","author":"Harper","year":"2014","journal-title":"Antibiotics"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1111\/joim.12782","article-title":"Biofilm formation\u2014What we can learn from recent developments","volume":"284","author":"Bjarnsholt","year":"2018","journal-title":"J. Intern Med."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Harper, D., Abedon, S., Burrowes, B., and McConville, M. (2019). Biofilm Applications of Bacteriophages. Bacteriophages, Springer.","DOI":"10.1007\/978-3-319-40598-8"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Woods, P.W., Haynes, Z.M., Mina, E.G., and Marques, C.N.H. (2019). Maintenance of S. aureus in Co-culture With P. aeruginosa While Growing as Biofilms. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.03291"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e00111-25","DOI":"10.1128\/msystems.00111-25","article-title":"Proteomic study of evolved Pseudomonas aeruginosa strains grown in Staphylococcus aureus- and Klebsiella pneumoniae-conditioned media","volume":"10","author":"Pan","year":"2025","journal-title":"mSystems"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4112","DOI":"10.1128\/IAI.00521-13","article-title":"MrkD1P from Klebsiella pneumoniae Strain IA565 Allows for Coexistence with Pseudomonas aeruginosa and Protection from Protease-Mediated Biofilm Detachment","volume":"81","author":"Childers","year":"2013","journal-title":"Infect. Immun."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1038\/s41522-024-00546-0","article-title":"Klebsiella pneumoniae AI-2 transporters mediate interspecies interactions and composition in a three-species biofilm community","volume":"10","author":"Subramoni","year":"2024","journal-title":"npj Biofilms Microbiomes"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1038\/nrmicro821","article-title":"Bacterial biofilms: From the Natural environment to infectious diseases","volume":"2","author":"Costerton","year":"2004","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Camus, L., Briaud, P., Vandenesch, F., and Moreau, K. (2021). How Bacterial Adaptation to Cystic Fibrosis Environment Shapes Interactions Between Pseudomonas aeruginosa and Staphylococcus aureus. Front Microbiol., 12.","DOI":"10.3389\/fmicb.2021.617784"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1259\/bjr\/28999734","article-title":"Acute Klebsiella pneumoniae pneumonia alone and with concurrent infection: Comparison of clinical and thin-section CT findings","volume":"83","author":"Okada","year":"2010","journal-title":"Br. J. Radiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1111\/j.1469-0691.2012.04001.x","article-title":"The polymicrobial nature of biofilm infection","volume":"19","author":"Wolcott","year":"2013","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1016\/j.tim.2014.05.007","article-title":"Polybacterial human disease: The ills of social networking","volume":"22","author":"Short","year":"2014","journal-title":"Trends Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Viksne, R., Racenis, K., Broks, R., Balode, A.O., Kise, L., and Kroica, J. (2023). In Vitro Assessment of Biofilm Production, Antibacterial Resistance of Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter spp. Obtained from Tonsillar Crypts of Healthy Adults. Microorganisms, 11.","DOI":"10.3390\/microorganisms11020258"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1016\/S0140-6736(21)02724-0","article-title":"Global burden of bacterial antimicrobial resistance in 2019: A systematic analysis","volume":"399","author":"Murray","year":"2022","journal-title":"Lancet"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Beaume, M., K\u00f6hler, T., Fontana, T., Tognon, M., Renzoni, A., and van Delden, C. (2015). Metabolic pathways of Pseudomonas aeruginosa involved in competition with respiratory bacterial pathogens. Front Microbiol., 6.","DOI":"10.3389\/fmicb.2015.00321"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1007\/s12275-014-4067-3","article-title":"Mechanisms of synergy in polymicrobial infections","volume":"52","author":"Murray","year":"2014","journal-title":"J. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1038\/s41392-022-01056-1","article-title":"Pseudomonas aeruginosa: Pathogenesis, virulence factors, antibiotic resistance, interaction with host, technology advances and emerging therapeutics","volume":"7","author":"Qin","year":"2022","journal-title":"Sig. Transduct. Target. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.1016\/S1473-3099(25)00118-5","article-title":"The WHO Bacterial Priority Pathogens List 2024: A prioritization study to guide research, development, and public health strategies against antimicrobial resistance","volume":"25","author":"Sati","year":"2025","journal-title":"Lancet Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e0060724","DOI":"10.1128\/msystems.00607-24","article-title":"Antibacterial effect of phage cocktails and phage-antibiotic synergy against pathogenic Klebsiella pneumoniae","volume":"9","author":"Zhao","year":"2024","journal-title":"mSystems"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Forti, F., Roach, D.R., Cafora, M., Pasini, M.E., Horner, D.S., Fiscarelli, E.V., Rossitto, M., Cariani, L., Briani, F., and Debarbieux, L. (2018). Design of a Broad-Range Bacteriophage Cocktail That Reduces Pseudomonas aeruginosa Biofilms and Treats Acute Infections in Two Animal Models. Antimicrob. Agents Chemother., 62.","DOI":"10.1128\/AAC.02573-17"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1376","DOI":"10.1039\/D5PM00094G","article-title":"Biofilms: Structure, resistance mechanism, emerging control strategies, and applications","volume":"2","author":"Ugwu","year":"2025","journal-title":"RSC Pharm."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"162","DOI":"10.4292\/wjgpt.v8.i3.162","article-title":"Phage therapy: An alternative to antibiotics in the age of multi-drug resistance","volume":"8","author":"Lin","year":"2017","journal-title":"World J. Gastrointest. Pharmacol. Ther."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"66","DOI":"10.4161\/bact.1.2.15845","article-title":"Phage treatment of human infections","volume":"1","author":"Abedon","year":"2011","journal-title":"Bacteriophage"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"769","DOI":"10.2217\/fmb.13.47","article-title":"Phage cocktails and the future of phage therapy","volume":"8","author":"Chan","year":"2013","journal-title":"Future Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Cooper, C.J., Mirzaei, M.K., and Nilsson, A.S. (2016). Adapting drug approval pathways for bacteriophage-based therapeutics. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.01209"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1434","DOI":"10.1038\/s41564-024-01705-x","article-title":"Personalized bacteriophage therapy outcomes for 100 consecutive cases: A multicentre, multinational, retrospective observational study","volume":"9","author":"Pirnay","year":"2024","journal-title":"Nat. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e00954-17","DOI":"10.1128\/AAC.00954-17","article-title":"Development and use of personalized bacteriophage-based therapeutic cocktails to treat a patient with a disseminated resistant Acinetobacter baumannii infection","volume":"61","author":"Schooley","year":"2017","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e01462-20","DOI":"10.1128\/mBio.01462-20","article-title":"Phage-antibiotic synergy is driven by a unique combination of antibacterial mechanism of action and stoichiometry","volume":"11","author":"Green","year":"2020","journal-title":"mBio"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Abedon, S.T. (2019). Phage-antibiotic combination treatments: Antagonistic impacts of antibiotics on the pharmacodynamics of phage therapy?. Antibiotics, 8.","DOI":"10.3390\/antibiotics8040182"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Tagliaferri, T.L., Jansen, M., and Horz, H.P. (2019). Fighting Pathogenic Bacteria on Two Fronts: Phages and Antibiotics as Combined Strategy. Front Cell Infect. Microbiol., 9.","DOI":"10.3389\/fcimb.2019.00022"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"109145","DOI":"10.1016\/j.lwt.2020.109145","article-title":"Characterization of two novel lytic bacteriophages for reducing biofilms of zoonotic multidrug-resistant Staphylococcus aureus and controlling their growth in milk","volume":"124","author":"Gharieb","year":"2020","journal-title":"LWT"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Pearl, S., Gabay, C., Kishony, R., Oppenheim, A., and Balaban, N.Q. (2008). Nongenetic individuality in the host-phage interaction. PLoS Biol., 6.","DOI":"10.1371\/journal.pbio.0060120"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"EMBR202357309","DOI":"10.15252\/embr.202357309","article-title":"Environmental, mechanistic and evolutionary landscape of antibiotic persistence","volume":"24","author":"Bollen","year":"2023","journal-title":"EMBO Rep."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Akturk, E., Oliveira, H., Santos, S.B., Costa, S., Kuyumcu, S., Melo, L.D.R., and Azeredo, J. (2019). Synergistic action of phage and antibiotics: Parameters to enhance the killing efficacy against mono and dual-species biofilms. Antibiotics, 8.","DOI":"10.3390\/antibiotics8030103"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1186\/s12941-020-00389-5","article-title":"Bacteriophage therapy against Pseudomonas aeruginosa biofilms: A review","volume":"19","author":"Chegini","year":"2020","journal-title":"Ann. Clin. Microbiol. Antimicrob."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1208\/s12248-019-0315-0","article-title":"Bacteriophage PEV20 and ciprofloxacin combination treatment enhances removal of Pseudomonas aeruginosa biofilm isolated from cystic fibrosis and wound patients","volume":"21","author":"Chang","year":"2019","journal-title":"AAPS J."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.ijpharm.2018.09.024","article-title":"Synergy of nebulized phage PEV20 and ciprofloxacin combination against Pseudomonas aeruginosa","volume":"551","author":"Lin","year":"2018","journal-title":"Int. J. Pharm."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3778","DOI":"10.3390\/v6103778","article-title":"Effect of bacteriophage infection in combination with tobramycin on the emergence of resistance in Escherichia coli and Pseudomonas aeruginosa biofilms","volume":"6","author":"Coulter","year":"2014","journal-title":"Viruses"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Kumaran, D., Taha, M., Yi, Q., Ramirez-Arcos, S., Diallo, J.-S., Carli, A., and Abdelbary, H. (2018). Does treatment order matter? investigating the ability of bacteriophage to augment antibiotic activity against Staphylococcus aureus biofilms. Front Microbiol., 9.","DOI":"10.3389\/fmicb.2018.00127"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"117","DOI":"10.2106\/JBJS.K.01135","article-title":"Bacteriophage therapy in implant-related infections an experimental study","volume":"95A","author":"Yilmaz","year":"2013","journal-title":"J. Bone Jt. Surg.-Am."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1128\/mBio.00029-12","article-title":"Bacteriophages phi MR299-2 and phi NH-4 can eliminate Pseudomonas aeruginosa in the murine lung and on cystic fibrosis lung airway cells","volume":"3","author":"Alemayehu","year":"2012","journal-title":"Mbio"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1111\/1751-7915.12316","article-title":"A novel bacteriophage cocktail reduces and disperses Pseudomonas aeruginosa biofilms under static and flow conditions","volume":"9","author":"Alves","year":"2016","journal-title":"Microb. Biotechnol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Racenis, K., Kroica, J., Rezevska, D., Avotins, L., Skuditis, E., Popova, A., Puide, I., Kuzema, V., and Petersons, A. (2020). S. aureus colonization, biofilm production, and phage susceptibility in peritoneal dialysis patients. Antibiotics, 9.","DOI":"10.3390\/antibiotics9090582"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Martinet, M.G., Thomas, M., Bojunga, J., Pletz, M.W., Vehreschild, M.J.G.T., and W\u00fcrstle, S. (2025). The landscape of biofilm models for phage therapy: Mimicking biofilms in diabetic foot ulcers using 3D models. Front Microbiol., 16.","DOI":"10.3389\/fmicb.2025.1553979"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2306116","DOI":"10.1002\/adfm.202306116","article-title":"Heterogeneity Governs 3D-Cultures of Clinically Relevant Microbial Communities","volume":"33","author":"Bertoglio","year":"2023","journal-title":"Adv. Funct. Mater."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Glonti, T., Goossens, M., Cochez, C., Green, S., Gorivale, S., Wagemans, J., Lavigne, R., and Pirnay, J.-P. (2024). Use of the Naturally Occurring Bacteriophage Grouping Model for the Design of Potent Therapeutic Cocktails. Antibiotics, 13.","DOI":"10.3390\/antibiotics13050385"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1111\/j.1751-7915.2011.00259.x","article-title":"Ealuation of lytic activity of staphylococcal bacteriophage Sb-1 against freshly isolated clinical pathogens","volume":"4","author":"Kvachadze","year":"2011","journal-title":"Microb. Biotechnol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.jgar.2024.01.021","article-title":"Evaluation of antibiofilm activity of cefiderocol alone and in combination with imipenem against Pseudomonas aeruginosa","volume":"37","author":"Ferretti","year":"2024","journal-title":"J. Glob. Antimicrob. Resist."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Abedon, S.T., Danis-Wlodarczyk, K.M., Wozniak, D.J., and Sullivan, M.B. (2021). Improving Phage-Biofilm In Vitro Experimentation. Viruses, 13.","DOI":"10.3390\/v13061175"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2321","DOI":"10.3168\/jds.2009-2800","article-title":"Microscopic observation of multispecies biofilm of various structures on whey concentration membranes","volume":"93","author":"Hassan","year":"2010","journal-title":"J. Dairy Sci."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"fnv246","DOI":"10.1093\/femsle\/fnv246","article-title":"Bacteriophage exploitation of bacterial biofilms: Phage preference for less mature targets?","volume":"363","author":"Abedon","year":"2016","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/j.meegid.2013.05.016","article-title":"Small colony variants (SCVs) of Staphylococcus aureus\u2014A bacterial survival strategy","volume":"21","author":"KahlB","year":"2014","journal-title":"Infect. Genet. Evol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"e946","DOI":"10.1002\/mbo3.946","article-title":"Biofilm formation and prevalence of adhesion genes among Staphylococcus aureus isolates from different food sources","volume":"9","author":"Chen","year":"2020","journal-title":"MicrobiologyOpen"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Kasetty, S., Katharios-Lanwermeyer, S., O\u2019Toole, G.A., and Nadell, C.D. (2021). Differential Surface Competition and Biofilm Invasion Strategies of Pseudomonas aeruginosa PA14 and PAO1. J. Bacteriol., 203.","DOI":"10.1128\/JB.00265-21"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Plotkin, B.J., Halkyard, S., Spoolstra, E., Micklo, A., Kaminski, A., Sigar, I.M., and Konaklieva, M.I. (2023). The Role of the Insulin\/Glucose Ratio in the Regulation of Pathogen Biofilm Formation. Biology, 12.","DOI":"10.3390\/biology12111432"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"623","DOI":"10.1038\/nrmicro2415","article-title":"The biofilm matrix","volume":"8","author":"Flemming","year":"2010","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.mib.2013.01.003","article-title":"Exploiting social evolution in biofilms","volume":"16","author":"Boyle","year":"2013","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1038\/s41522-017-0035-0","article-title":"Staphylococcus aureus interaction with Pseudomonas aeruginosa biofilm enhances tobramycin resistance","volume":"3","author":"Beaudoin","year":"2017","journal-title":"npj Biofilms Microbiomes"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"19890","DOI":"10.1073\/pnas.0606756104","article-title":"Selection for Staphylococcus aureus small-colony variants due to growth in the presence of Pseudomonas aeruginosa","volume":"103","author":"Hoffman","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.molcel.2018.10.022","article-title":"ATP-Dependent Dynamic Protein Aggregation Regulates Bacterial Dormancy Depth Critical for Antibiotic Tolerance","volume":"73","author":"Pu","year":"2019","journal-title":"Mol. Cell"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"102984","DOI":"10.1016\/j.xpro.2024.102984","article-title":"Protocol for assessing single-cell persister recovery kinetics and physiology in Escherichia coli using spectrophotometry","volume":"5","author":"Nguyen","year":"2024","journal-title":"STAR Protoc."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"S39","DOI":"10.1016\/S1569-1993(10)60153-1","article-title":"An ex vivo sputum model to evaluate antibacterial treatments in CF patients chronically colonized with Pseudomonas aeruginosa","volume":"9","author":"Glonti","year":"2010","journal-title":"J. Cyst. Fibros."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Salcedo-Sora, J.E., and Kell, D.B. (2020). A Quantitative Survey of Bacterial Persistence in the Presence of Antibiotics: Towards Antipersister Antimicrobial Discovery. Antibiotics, 9.","DOI":"10.3390\/antibiotics9080508"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"EMBR202051034","DOI":"10.15252\/embr.202051034","article-title":"Antibiotic resistance and persistence\u2014Implications for human health and treatment perspectives","volume":"21","author":"Huemer","year":"2020","journal-title":"EMBO Rep."},{"key":"ref_65","first-page":"e0114925","article-title":"Toroidal displacement of Klebsiella pneumoniae by Pseudomonas aeruginosa is a unique mechanism to avoid competition for iron","volume":"16","author":"Pradhan","year":"2022","journal-title":"bioRxiv"},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Luo, Z.X., Li, Y., Liu, M.F., and Zhao, R. (2023). Ciprofloxacin enhances the biofilm formation of Staphylococcus aureus via an agrC-dependent mechanism. Front Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1328947"},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Engeman, E., Freyberger, H.R., Corey, B.W., Ward, A.M., He, Y., Nikolich, M.P., Filippov, A.A., Tyner, S.D., and Jacobs, A.C. (2021). Synergistic Killing and Re-Sensitization of Pseudomonas aeruginosa to Antibiotics by Phage-Antibiotic Combination Treatment. Pharmaceuticals, 14.","DOI":"10.3390\/ph14030184"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"e00143-21","DOI":"10.1128\/AAC.00143-21","article-title":"Lytic bacteriophages facilitate antibiotic sensitization of Enterococcus faecium","volume":"65","author":"Canfield","year":"2021","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_69","unstructured":"Shiffman, M., and Low, M. (2018). Bacteriophage-Mediated Biocontrol of Wound Infections, and Ecological Exploitation of Biofilms by Phages. Biofilm, Pilonidal Cysts and Sinuses. Recent Clinical Techniques, Results, and Research in Wounds, Springer."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"2299","DOI":"10.1038\/s41467-025-57508-z","article-title":"Antibiotic-mediated microbial community restructuring is dictated by variability in antibiotic-induced lysis rates and population interactions","volume":"16","author":"Kim","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"4585","DOI":"10.1128\/JB.185.15.4585-4592.2003","article-title":"Cell death in Pseudomonas aeruginosa biofilm development","volume":"185","author":"Webb","year":"2003","journal-title":"J. Bacteriol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1038\/s44259-024-00051-6","article-title":"Antibiotic susceptibility testing using minimum inhibitory concentration (MIC) assays","volume":"2","author":"Mahmood","year":"2024","journal-title":"npj Antimicrob. Resist"},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Glonti, T., and Pirnay, J.-P. (2022). In Vitro Techniques and Measurements of Phage Characteristics That Are Important for Phage Therapy Success. 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