{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T23:17:56Z","timestamp":1792883876347,"version":"4.5.0"},"reference-count":50,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,2,20]],"date-time":"2023-02-20T00:00:00Z","timestamp":1676851200000},"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":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Multidrug-resistant (MDR) pathogens are being recognized as a critical threat to human health if they can form biofilm and, in this sense, biofilm-forming MDR-methicillin resistant\n                      <jats:italic>Staphylococcus aureus<\/jats:italic>\n                      (MRSA) and -\n                      <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                      strains are a worse concern. Hence, a growing body of documents has introduced antimicrobial peptides (AMPs) as a substitute candidate for conventional antimicrobial agents against drug-resistant and biofilm-associated infections. We evaluated melittin\u2019s antibacterial and antibiofilm activity alone and\/or in combination with gentamicin, ciprofloxacin, rifampin, and vancomycin on biofilm-forming MDR-\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      and MDR-MRSA strains.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Antibacterial tests [antibiogram, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC)], anti-biofilm tests [minimum biofilm inhibition concentration (MBIC), and minimum biofilm eradication concentration (MBEC)], as well as synergistic antibiofilm activity of melittin and antibiotics, were performed. Besides, the influence of melittin alone on the biofilm encoding genes and the cytotoxicity and hemolytic effects of melittin were examined.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      MIC, MBC, MBIC, and MBEC indices for melittin were in the range of 0.625\u20135, 1.25\u201310, 2.5\u201320, and 10\u201340 \u03bcg\/ml, respectively. The findings found that the combination of melittin AMP with antibiotics was synergistic and fractional biofilm inhibitory concentration index (FBICi) for most tested concentrations was &amp;lt;0.5, resulting in a significant reduction in melittin, gentamicin, ciprofloxacin, vancomycin, and rifampin concentrations by 2\u2013256.4, 2\u2013128, 2\u201316, 4\u201364 and 4\u20138 folds, respectively. This phenomenon reduced the toxicity of melittin, whereby its synergist concentration required for biofilm inhibition did not show cytotoxicity and hemolytic activity. Our findings found that melittin decreased the expression of\n                      <jats:italic>icaA<\/jats:italic>\n                      in\n                      <jats:italic>S. aureus<\/jats:italic>\n                      and\n                      <jats:italic>LasR<\/jats:italic>\n                      in\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      genes from 0.1 to 4.11 fold for\n                      <jats:italic>icaA<\/jats:italic>\n                      , and 0.11 to 3.7 fold for\n                      <jats:italic>LasR<\/jats:italic>\n                      , respectively.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Overall, the results obtained from our study show that melittin alone is effective against the strong biofilm of MDR pathogens and also offers sound synergistic effects with antibiotics without toxicity. Hence, combining melittin and antibiotics can be a potential candidate for further evaluation of\n                      <jats:italic>in vivo<\/jats:italic>\n                      infections by MDR pathogens.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2023.1030401","type":"journal-article","created":{"date-parts":[[2023,2,20]],"date-time":"2023-02-20T00:17:15Z","timestamp":1676852235000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":40,"title":["Antibiofilm effect of melittin alone and in combination with conventional antibiotics toward strong biofilm of MDR-MRSA and -Pseudomonas aeruginosa"],"prefix":"10.3389","volume":"14","author":[{"given":"Rasoul","family":"Mirzaei","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hadi","family":"Esmaeili Gouvarchin Ghaleh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reza","family":"Ranjbar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,2,20]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"100646","DOI":"10.1016\/j.mgene.2020.100646","article-title":"Detection of biofilm formation and assessment of biofilm genes expression in different Pseudomonas aeruginosa clinical isolates","volume":"23","author":"Abdelraheem","year":"2020","journal-title":"Meta Gene"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"1275","DOI":"10.1007\/s00726-022-03180-2","article-title":"Fast killing kinetics, significant therapeutic index, and high stability of melittin-derived antimicrobial peptide","volume":"54","author":"Akbari","year":"2022","journal-title":"Amino Acids"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1089\/mdr.2018.0016","article-title":"Highly synergistic effects of melittin with conventional antibiotics against multidrug-resistant isolates of acinetobacter baumannii and pseudomonas aeruginosa","volume":"25","author":"Akbari","year":"2019","journal-title":"Microb. Drug Resist."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1186\/s40360-021-00503-z","article-title":"In vitro and in vivo toxicity and antibacterial efficacy of melittin against clinical extensively drug-resistant bacteria","volume":"22","author":"Askari","year":"2021","journal-title":"BMC Pharmacol. Toxicol."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2012\/976972","article-title":"Prevalence of adhesion and regulation of biofilm-related genes in different clones of Staphylococcus aureus","volume":"2012","author":"Atshan","year":"2012","journal-title":"J. Biomed. Biotechnol."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"e0227927","DOI":"10.1371\/journal.pone.0227927","article-title":"Selection and validation of reference genes for gene expression studies in pseudomonas brassicacearum GS20 using real-time quantitative reverse transcription PCR","volume":"15","author":"Bai","year":"2020","journal-title":"PLoS One"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"443","DOI":"10.1007\/s10096-018-3189-7","article-title":"Highly synergistic activity of melittin with imipenem and colistin in biofilm inhibition against multidrug-resistant strong biofilm producer strains of Acinetobacter baumannii","volume":"37","author":"Bardbari","year":"2018","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"iv23","DOI":"10.1093\/jac\/dkab352","article-title":"Current and future perspectives in the treatment of multidrug-resistant gram-negative infections","volume":"76","author":"Bassetti","year":"2021","journal-title":"J. Antimicrob. Chemother."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/j.toxicon.2021.05.015","article-title":"Eradication of vancomycin-resistant Staphylococcus aureus on a mouse model of third-degree burn infection by melittin: an antimicrobial peptide from bee venom","volume":"199","author":"Bevalian","year":"2021","journal-title":"Toxicon"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"6483","DOI":"10.3892\/mmr.2015.4275","article-title":"Melittin, a honeybee venom-derived antimicrobial peptide, may target methicillin-resistant Staphylococcus aureus","volume":"12","author":"Choi","year":"2015","journal-title":"Mol. Med. Rep."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/srep23347","article-title":"Attenuation of Pseudomonas aeruginosa biofilm formation by Vitexin: a combinatorial study with azithromycin and gentamicin","volume":"6","author":"Das","year":"2016","journal-title":"Sci. Rep."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"S113","DOI":"10.1016\/j.jcot.2019.10.002","article-title":"Effects of powdered rifampin and vancomycin solutions on biofilm production of staphylococcus aureus on orthopedic implants","volume":"11","author":"Douthit","year":"2020","journal-title":"J. clinical orthopaedics and trauma"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"687","DOI":"10.1111\/cbdd.12702","article-title":"A new, high yield, rapid, and cost-effective protocol to deprotection of cysteine-rich conopeptide, omega-conotoxin MVIIA","volume":"87","author":"Eisapoor","year":"2016","journal-title":"Chem. Biol. Drug Des."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1002\/ppul.10127","article-title":"Pseudomonas aeruginosa and other predictors of mortality and morbidity in young children with cystic fibrosis","volume":"34","author":"Emerson","year":"2002","journal-title":"Pediatr. Pulmonol."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"430","DOI":"10.1093\/femsre\/fux007","article-title":"Antibiotic resistance in Staphylococcus aureus. Current status and future prospects","volume":"41","author":"Foster","year":"2017","journal-title":"FEMS Microbiol. Rev."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"951","DOI":"10.1586\/14787210.5.6.951","article-title":"Alternative mechanisms of action of cationic antimicrobial peptides on bacteria","volume":"5","author":"Hale","year":"2007","journal-title":"Expert Rev. Anti-Infect. Ther."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"593","DOI":"10.1016\/j.jiac.2014.08.001","article-title":"Multi-drug-resistant Staphylococcus aureus and future chemotherapy","volume":"20","author":"Hiramatsu","year":"2014","journal-title":"J. Infect. Chemother."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"309","DOI":"10.1016\/j.ajic.2008.03.002","article-title":"CDC\/NHSN surveillance definition of health care\u2013associated infection and criteria for specific types of infections in the acute care setting","volume":"36","author":"Horan","year":"2008","journal-title":"Am. J. Infect. Control"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"00031-19","DOI":"10.1128\/CMR.00031-19","article-title":"Epidemiology and treatment of multidrug-resistant and extensively drug-resistant Pseudomonas aeruginosa infections","volume":"32","author":"Horcajada","year":"2019","journal-title":"Clin. Microbiol. Rev."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1128\/CMR.00042-09","article-title":"Reduced vancomycin susceptibility in Staphylococcus aureus, including vancomycin-intermediate and heterogeneous vancomycin-intermediate strains: resistance mechanisms, laboratory detection, and clinical implications","volume":"23","author":"Howden","year":"2010","journal-title":"Clin. Microbiol. Rev."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"2033","DOI":"10.1128\/JCM.00196-13","article-title":"Detection of methicillin-resistant Staphylococcus aureus by a duplex droplet digital PCR assay","volume":"51","author":"Kelley","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1007\/s10989-018-9675-z","article-title":"Kinetics study of antimicrobial peptide, melittin, in simultaneous biofilm degradation and eradication of potent biofilm producing MDR Pseudomonas aeruginosa isolates","volume":"25","author":"Khozani","year":"2019","journal-title":"Int. J. Pept. Res. Ther."},{"key":"ref23","doi-asserted-by":"publisher","first-page":"1","DOI":"10.18869\/acadpub.mlj.10.1.1","article-title":"Association of agr gene expression with Staphylococcus aureus virulence genes in BHI broth","volume":"10","author":"Koohsari","year":"2016","journal-title":"Medical Lab. J."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"18033","DOI":"10.1038\/nrdp.2018.33","article-title":"Methicillin-resistant Staphylococcus aureus","volume":"4","author":"Lee","year":"2018","journal-title":"Nat. Rev. Dis. 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Microbiol."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"869650","DOI":"10.3389\/fmicb.2022.869650","article-title":"Highly synergistic effects of Melittin with vancomycin and rifampin against vancomycin and rifampin resistant Staphylococcus epidermidis","volume":"13","author":"Mirzaei","year":"","journal-title":"Front. Microbiol."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"112670","DOI":"10.1016\/j.biopha.2022.112670","article-title":"Prevention, inhibition, and degradation effects of melittin alone and in combination with vancomycin and rifampin against strong biofilm producer strains of methicillin-resistant Staphylococcus epidermidis","volume":"147","author":"Mirzaei","year":"","journal-title":"Biomed. Pharmacother."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"1271","DOI":"10.1002\/iub.2266","article-title":"The biofilm-associated bacterial infections unrelated to indwelling devices","volume":"72","author":"Mirzaei","year":"","journal-title":"IUBMB Life"},{"key":"ref34","doi-asserted-by":"publisher","first-page":"104393","DOI":"10.1016\/j.micpath.2020.104393","article-title":"The importance of intracellular bacterial biofilm in infectious diseases","volume":"147","author":"Mirzaei","year":"","journal-title":"Microb. Pathog."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"108471","DOI":"10.1016\/j.intimp.2021.108471","article-title":"Hijacking host components for bacterial biofilm formation: an advanced mechanism","volume":"103","author":"Mirzaei","year":"2022","journal-title":"Int. Immunopharmacol."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"e0277287","DOI":"10.1371\/journal.pone.0277287","article-title":"The issue beyond resistance: methicillin-resistant Staphylococcus epidermidis biofilm formation is induced by subinhibitory concentrations of cloxacillin, cefazolin, and clindamycin","volume":"17","author":"Mirzaei","year":"","journal-title":"PLoS One"},{"key":"ref37","first-page":"449","article-title":"Manual of clinical microbiology","volume":"3","author":"Murray","year":"1995","journal-title":"Trends Microbiol."},{"key":"ref38","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1016\/j.biotechadv.2018.11.013","article-title":"Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies","volume":"37","author":"Pang","year":"2019","journal-title":"Biotechnol. Adv."},{"key":"ref39","first-page":"71","article-title":"An improvement of the 2\u02c6 (\u2212delta delta CT) method for quantitative real-time polymerase chain reaction data analysis","volume":"3","author":"Rao","year":"2013","journal-title":"Biostat., bioinfo. biomathemat."},{"key":"ref40","volume-title":"Anti-biofilm effect of Melittin peptide on clinical isolates of pseudomonas aeruginosa isolated from hospital burn infections","author":"Shams","year":"2020"},{"key":"ref41","doi-asserted-by":"publisher","first-page":"543","DOI":"10.3389\/fmicb.2019.00543","article-title":"Biofilm forming antibiotic resistant gram-positive pathogens isolated from surfaces on the international space station","volume":"10","author":"Sobisch","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref42","doi-asserted-by":"publisher","DOI":"10.3390\/metabo10050191","article-title":"The effect of bee venom peptides Melittin, Tertiapin, and Apamin on the human erythrocytes ghosts: a preliminary study","volume":"10","author":"\u015awiat\u0142y-B\u0142aszkiewicz","year":"2020","journal-title":"Meta"},{"key":"ref43","doi-asserted-by":"publisher","first-page":"1735","DOI":"10.1128\/AAC.01022-08","article-title":"Inhibitory and bactericidal activities of daptomycin, vancomycin, and teicoplanin against methicillin-resistant Staphylococcus aureus isolates collected from 1985 to 2007","volume":"53","author":"Traczewski","year":"2009","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"377","DOI":"10.1016\/j.idc.2016.02.004","article-title":"Multidrug-resistant bacteria in the community: trends and lessons learned","volume":"30","author":"Van Duin","year":"2016","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref45","doi-asserted-by":"publisher","first-page":"709","DOI":"10.1016\/j.idc.2020.08.002","article-title":"Multidrug-resistant bacteria in the community: an update","volume":"34","author":"Van Duin","year":"2020","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref46","first-page":"M100","article-title":"Clinical and laboratory standards institute: performance standards for antimicrobial susceptibility testing: 20th informational supplement","author":"Wayne","year":"2010","journal-title":"CLSI document"},{"key":"ref47","doi-asserted-by":"publisher","first-page":"124432","DOI":"10.1016\/j.matchemphys.2021.124432","article-title":"Melittin antimicrobial peptide thin layer on bone implant chitosan-antibiotic coatings and their bactericidal properties","volume":"263","author":"Zarghami","year":"","journal-title":"Mater. Chem. Phys."},{"key":"ref48","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s10856-021-06551-5","article-title":"Prevention the formation of biofilm on orthopedic implants by melittin thin layer on chitosan\/bioactive glass\/vancomycin coatings","volume":"32","author":"Zarghami","year":"","journal-title":"J. Mater. Sci. Mater. Med."},{"key":"ref49","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s10856-022-06666-3","article-title":"Improving bactericidal performance of implant composite coatings by synergism between Melittin and tetracycline","volume":"33","author":"Zarghami","year":"2022","journal-title":"J. Mater. Sci. Mater. Med."},{"key":"ref50","doi-asserted-by":"publisher","first-page":"563","DOI":"10.1007\/s10989-017-9641-1","article-title":"Apoptotic effect of melittin purified from Iranian honey bee venom on human cervical cancer HeLa cell line","volume":"24","author":"Zarrinnahad","year":"2018","journal-title":"Int. J. Pept. Res. 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