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scholarship","award":["#ISF1349\/20"],"award-info":[{"award-number":["#ISF1349\/20"]}]},{"name":"The Foulks Foundation scholarship","award":["A2232"],"award-info":[{"award-number":["A2232"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Background: Antibiotic-resistant Pseudomonas aeruginosa (P. aeruginosa) strains are an increasing cause of morbidity and mortality. Pulsed blue light (PBL) enhances porphyrin-induced reactive oxygen species and has been clinically shown to be harmless to the skin at low doses. Bacteriophages, viruses that infect bacteria, offer a promising non-antibiotic bactericidal approach. This study investigates the potential synergism between low-dose PBL and phage therapy against P. aeruginosa in planktonic cultures and preformed biofilms. Methods: We conducted a factorial dose\u2013response in vitro study combining P. aeruginosa-specific phages with PBL (457 nm, 33 kHz) on both PA14 and multidrug-resistant PATZ2 strains. After excluding direct PBL effects on phage titer or activity, we assessed effectiveness on planktonic cultures using growth curve analysis (via growth_curve_outcomes, a newly developed, Python-based tool available on GitHub) , CFU, and PFU. Biofilm efficacy was evaluated using CFU post-sonication, crystal violet staining, and live\/dead staining with confocal microscopy. Finally, we assessed reactive oxygen species (ROS) as a potential mechanism using the nitro blue tetrazolium reduction assay. ANOVA or Kruskal\u2013Wallis tests with post hoc Tukey or Conover\u2013Iman tests were used for comparisons (n = 5 biological replicates and technical triplicates). Results: The bacterial growth lag phase was significantly extended for phage alone or PBL alone, with a synergistic effect of up to 144% (p &lt; 0.001 for all), achieving a 9 log CFU\/mL reduction at 24 h (p &lt; 0.001). In preformed biofilms, synergistic combinations significantly reduced biofilm biomass and bacterial viability (% Live, median (IQR): Control 80%; Phage 40%; PBL 25%; PBL&amp;Phage 15%, p &lt; 0.001). Mechanistically, PBL triggered transient ROS in planktonic cultures, amplified by phage co-treatment, while a biphasic ROS pattern in biofilms reflected time-dependent synergy. Conclusions: Phage therapy combined with PBL demonstrates a synergistic bactericidal effect against P. aeruginosa in both planktonic cultures and biofilms. Given the strong safety profile of PBL and phages, this approach may lead to a novel, antibiotic-complementary, safe treatment modality for patients suffering from difficult-to-treat antibiotic-resistant infections and biofilm-associated infections.<\/jats:p>","DOI":"10.3390\/antibiotics14050481","type":"journal-article","created":{"date-parts":[[2025,5,9]],"date-time":"2025-05-09T08:20:23Z","timestamp":1746778823000},"page":"481","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Pulsed Blue Light and Phage Therapy: A Novel Synergistic Bactericide"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1749-4786","authenticated-orcid":false,"given":"Amit","family":"Rimon","sequence":"first","affiliation":[{"name":"Institute of Biomedical and Oral Research (IBOR), Faculty of Dental Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"},{"name":"Tzameret, The Military Track of Medicine, The Hebrew University-Hadassah Medical School, Jerusalem 9112001, Israel"},{"name":"The Israeli Phage Therapy Center (IPTC) of the Hebrew University and Hadassah Medical Center, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-7363-4004","authenticated-orcid":false,"given":"Jonathan","family":"Belin","sequence":"additional","affiliation":[{"name":"Institute of Biomedical and Oral Research (IBOR), Faculty of Dental Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ortal","family":"Yerushalmy","sequence":"additional","affiliation":[{"name":"Institute of Biomedical and Oral Research (IBOR), Faculty of Dental Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"},{"name":"The Israeli Phage Therapy Center (IPTC) of the Hebrew University and Hadassah Medical Center, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yonatan","family":"Eavri","sequence":"additional","affiliation":[{"name":"Institute for Research in Military Medicine (IRMM), Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anatoly","family":"Shapochnikov","sequence":"additional","affiliation":[{"name":"The Department of Medical Neurobiology, Institute for Medical Research (IMRIC), Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shunit","family":"Coppenhagen-Glazer","sequence":"additional","affiliation":[{"name":"Institute of Biomedical and Oral Research (IBOR), Faculty of Dental Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"},{"name":"The Israeli Phage Therapy Center (IPTC) of the Hebrew University and Hadassah Medical Center, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9967-730X","authenticated-orcid":false,"given":"Ronen","family":"Hazan","sequence":"additional","affiliation":[{"name":"Institute of Biomedical and Oral Research (IBOR), Faculty of Dental Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"},{"name":"The Israeli Phage Therapy Center (IPTC) of the Hebrew University and Hadassah Medical Center, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0392-8958","authenticated-orcid":false,"given":"Lilach","family":"Gavish","sequence":"additional","affiliation":[{"name":"Institute for Research in Military Medicine (IRMM), Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, Israel"},{"name":"The Department of Medical Neurobiology, Institute for Medical Research (IMRIC), Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, Israel"},{"name":"The Saul and Joyce Brandman Hub for Cardiovascular Research, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,9]]},"reference":[{"key":"ref_1","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_2","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1016\/S1473-3099(16)00065-7","article-title":"Current Evidence on Hospital Antimicrobial Stewardship Objectives: A Systematic Review and Meta-Analysis","volume":"16","author":"Schuts","year":"2016","journal-title":"Lancet Infect. 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