{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T13:04:14Z","timestamp":1790168654903,"version":"4.1.0"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,1,17]],"date-time":"2024-01-17T00:00:00Z","timestamp":1705449600000},"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. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Bacteriophage therapy is becoming part of mainstream Western medicine since antibiotics of clinical use tend to fail. It involves applying lytic bacteriophages that self-replicate and induce cell lysis, thus killing their hosts. Nevertheless, bacterial killing promotes the selection of resistant clones which sometimes may exhibit a decrease in bacterial virulence or antibiotic resistance.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In this work, we studied the <jats:italic>Pseudomonas aeruginosa<\/jats:italic> lytic phage \u03c6DCL-PA6 and its variant \u03c6DCL-PA6\u03b1. Additionally, we characterized and evaluated the production of virulence factors and the virulence in a Galleria mellonella model of resistant mutants against each phage for PA14 and two clinical strains.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Phage \u03c6DCL-PA6\u03b1 differs from the original by only two amino acids: one in the baseplate wedge subunit and another in the tail fiber protein. According to genomic data and cross-resistance experiments, these changes may promote the change of the phage receptor from the O-antigen to the core lipopolysaccharide. Interestingly, the host range of the two phages differs as determined against the <jats:italic>Pseudomonas aeruginosa<\/jats:italic> reference strains PA14 and PAO1 and against nine multidrug-resistant isolates from ventilator associated pneumonia.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>We show as well that phage resistance impacts virulence factor production. Specifically, phage resistance led to decreased biofilm formation, swarming, and type III secretion; therefore, the virulence towards Galleria mellonella was dramatically attenuated. Furthermore, antibiotic resistance decreased for one clinical strain. Our study highlights important potential advantages of phage therapy\u2019s evolutionary impact that may be exploited to generate robust therapy schemes.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2023.1280265","type":"journal-article","created":{"date-parts":[[2024,1,17]],"date-time":"2024-01-17T04:34:10Z","timestamp":1705466050000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":22,"title":["Resistance against two lytic phage variants attenuates virulence and antibiotic resistance in Pseudomonas aeruginosa"],"prefix":"10.3389","volume":"13","author":[{"given":"Juan Carlos","family":"Garc\u00eda-Cruz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xareni","family":"Rebollar-Juarez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aldo","family":"Limones-Martinez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristian Sadalis","family":"Santos-Lopez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shotaro","family":"Toya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshinari","family":"Maeda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Corina Diana","family":"Ceap\u0103","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucia","family":"Blasco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda","family":"Tom\u00e1s","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clara Estela","family":"D\u00edaz-Vel\u00e1squez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felipe","family":"Vaca-Paniagua","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miguel","family":"D\u00edaz-Guerrero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Cazares","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adri\u00e1n","family":"Cazares","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melisa","family":"Hern\u00e1ndez-Dur\u00e1n","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis Esa\u00fa","family":"L\u00f3pez-J\u00e1come","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael","family":"Franco-Cendejas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fohad Mabood","family":"Husain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Altaf","family":"Khan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammed","family":"Arshad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosario","family":"Morales-Espinosa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Mar\u00eda","family":"Fern\u00e1ndez-Presas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederic","family":"Cadet","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas K.","family":"Wood","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodolfo","family":"Garc\u00eda-Contreras","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,1,17]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local alignment search tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J. Mol. Biol."},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1089\/cmb.2012.0021","article-title":"SPAdes: A new genome assembly algorithm and its applications to single-cell sequencing","volume":"19","author":"Bankevich","year":"2012","journal-title":"J. Comput. Biol"},{"key":"B3","doi-asserted-by":"publisher","first-page":"4488","DOI":"10.1038\/s41598-022-08111-5","article-title":"The role of PemIK (PemK\/PemI) type II TA system from Klebsiella pneumoniae clinical strains in lytic phage infection","volume":"12","author":"Bleriot","year":"2022","journal-title":"Sci. Rep."},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu170","article-title":"Trimmomatic: A flexible trimmer for Illumina sequence data","volume":"30","author":"Bolger","year":"2014","journal-title":"Bioinformatics"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1038\/nature11723","article-title":"Bacteriophage genes that inactivate the CRISPR\/Cas bacterial immune system","volume":"493","author":"Bondy-Denomy","year":"2012","journal-title":"Nature"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2104592118","article-title":"Coevolutionary phage training leads to greater bacterial suppression and delays the evolution of phage resistance","volume":"118","author":"Borin","year":"2021","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.7554\/ELIFE.73679","article-title":"Parallel evolution of Pseudomonas aeruginosa phage resistance and virulence loss in response to phage treatment in vivo and in vitro","volume":"11","author":"Castledine","year":"2022","journal-title":"Elife"},{"key":"B8","doi-asserted-by":"publisher","first-page":"26717","DOI":"10.1038\/srep26717","article-title":"Phage selection restores antibiotic sensitivity in MDR Pseudomonas aerugin","volume":"6","author":"Chan","year":"2016","journal-title":"Sci. Rep."},{"key":"B9","volume-title":"Performance standards for antimicrobial Susceptibility Testing, 30th ed. CLSI supplement M100","year":"2020"},{"key":"B10","doi-asserted-by":"publisher","first-page":"884","DOI":"10.1128\/jb.172.2.884-900.1990","article-title":"Identification and characterization of genes for a second anthranilate synthase in Pseudomonas aeruginosa: Interchangeability of the two anthranilate synthase and evolutionary implications","volume":"172","author":"Essar","year":"1990","journal-title":"J. Bacteriol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"7084","DOI":"10.3390\/ijms23137084","article-title":"Flagellotropic bacteriophages: opportunities and challenges for antimicrobial applications","volume":"23","author":"Esteves","year":"2022","journal-title":"Int. J. Mol. Sci."},{"key":"B12","doi-asserted-by":"publisher","first-page":"246","DOI":"10.3390\/microorganisms9020246","article-title":"Taxonomic Evaluation of the Heyndrickxia (Basonym Bacillus) sporothermodurans Group (H. sporothermodurans, H. vini, H. oleronia) Based on Whole Genome Sequences","volume":"9","author":"Fiedler","year":"2021","journal-title":"Microorganisms"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1002\/cpbi.108","article-title":"Protein sequence analysis using the MPI bioinformatics toolkit","volume":"72","author":"Gabler","year":"2020","journal-title":"Curr. Protoc. Bioinforma."},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1093\/femsle\/fnaa080","article-title":"Rhamnolipids stabilize quorum sensing mediated cooperation in Pseudomonas aeruginosa","volume":"367","author":"Garc\u00eda-Contreras","year":"2020","journal-title":"FEMS Microbiol. Lett."},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00066-18","article-title":"Phage therapy in the postantibiotic era","volume":"32","author":"Gordillo Altamirano","year":"2019","journal-title":"Clin. Microbiol. Rev."},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-020-00830-7","article-title":"Bacteriophage-resistant Acinetobacter baumannii are resensitized to antimicrobials","volume":"6","author":"Gordillo Altamirano","year":"2021","journal-title":"Nat. Microbiol."},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2019.10.007","article-title":"Steering phages to combat bacterial pathogens","volume":"28","author":"Gurney","year":"2020","journal-title":"Trends Microbiol."},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2011.06.009","article-title":"Bacteriophages and their genomes","volume":"1","author":"Hatfull","year":"2011","journal-title":"Curr. Opin. Virol."},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00165-19","article-title":"Unique regions of the polysaccharide copolymerase wzz2 from pseudomonas aeruginosa are essential for o-specific antigen chain length control","volume":"201","author":"Huszczynski","year":"2019","journal-title":"J. Bacteriol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"6","DOI":"10.3390\/pathogens9010006","article-title":"The role of Pseudomonas aeruginosa lipopolysaccharide in bacterial pathogenesis and physiology","volume":"9","author":"Huszczynski","year":"2020","journal-title":"Pathogens"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1002\/bies.201400152","article-title":"A century of phage research: Bacteriophages and the shaping of modern biology","volume":"37","author":"Keen","year":"2015","journal-title":"BioEssays"},{"key":"B22","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1177\/1753425909106436","article-title":"Lipopolysaccharide biosynthesis in Pseudomonas aeruginosa","volume":"15","author":"King","year":"2009","journal-title":"Innate Immun."},{"key":"B23","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1007\/978-160327-164-6_14","article-title":"Phage host range and efficiency of plating","volume":"501","author":"Kutter","year":"2009","journal-title":"Methods Mol. Biol."},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.02072-22","article-title":"Characterization of phage resistance and their impacts on bacterial fitness in pseudomonas aeruginosa","volume":"10","author":"Li","year":"2022","journal-title":"Microbiol. Spectr."},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.02322","article-title":"Seeding public goods is essential for maintaining cooperation in pseudomonas aeruginosa","volume":"10","author":"Loarca","year":"2019","journal-title":"Front. Microbiol."},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1016\/j.clinthera.2020.07.014","article-title":"Phage therapy in the resistance era: where do we stand and where are we going","volume":"42","author":"Luong","year":"2020","journal-title":"Clin. Ther."},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00926-19","article-title":"Fitness trade-offs resulting from bacteriophage resistance potentiate synergistic antibacterial strategies","volume":"88","author":"Mangalea","year":"2020","journal-title":"Infect. Immun."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1332","DOI":"10.3390\/v13071332","article-title":"Emerging phage resistance in pseudomonas aeruginosa PAO1 is accompanied by an enhanced heterogeneity and reduced virulence","volume":"13","author":"Markwitz","year":"2021","journal-title":"Viruses"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1002\/ece3.9712","article-title":"Phage-resistant mutations impact bacteria susceptibility to future phage infections and antibiotic response","volume":"13","author":"McGee","year":"2023","journal-title":"Ecol. Evol."},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1002\/jobm.202200432","article-title":"Unraveling the regulation of pyocyanin synthesis by RsmA through MvaU and RpoS in Pseudomonas aeruginosa ID4365","volume":"63","author":"Montelongo-Mart\u00ednez","year":"2023","journal-title":"J. Basic Microbiol."},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.3791\/52338","article-title":"Preparation, imaging, and quantification of bacterial surface motility assays","volume":"2015","author":"Morales-Soto","year":"2015","journal-title":"J. Vis. Exp."},{"key":"B32","doi-asserted-by":"publisher","first-page":"D678","DOI":"10.1093\/nar\/gkac1003","article-title":"Introducing the Bacterial and Viral Bioinformatics Resource Center (BV-BRC): a resource combining PATRIC, IRD and ViPR","volume":"51","author":"Olson","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkac044","article-title":"Adaptation of clinical isolates of Klebsiella pneumoniae to the combination of niclosamide with the efflux pump inhibitor phenyl-arginine-\u03b2-naphthylamide (Pa\u03b2N): co-resistance to antimicrobials","volume":"77","author":"Pacios","year":"2022","journal-title":"J. Antimicrob. Chemother."},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-7343-9_16","article-title":"\u201cAnnotation of bacteriophage genome sequences using DNA master: An overview,\u201d","volume":"1681","author":"Pope","year":"2018","journal-title":"Methods Mol. Biol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky448","article-title":"HMMER web server: 2018 update","volume":"46","author":"Potter","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2020.12.011","article-title":"A phage-encoded anti-activator inhibits quorum sensing in Pseudomonas aeruginosa","volume":"81","author":"Shah","year":"2021","journal-title":"Mol Cell."},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1038\/s41396-021-00938-6","article-title":"Iron limitation by transferrin promotes simultaneous cheating of pyoverdine and exoprotease in Pseudomonas aeruginosa","volume":"15","author":"Tostado-Islas","year":"2021","journal-title":"ISME J"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics10020145","article-title":"Pseudomonas aeruginosa resistance to bacteriophages and its prevention by strategic therapeutic cocktail formulation","volume":"10","author":"Vaitekenas","year":"2021","journal-title":"Antibiotics"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2022.1004733","article-title":"Phage-resistant Pseudomonas aeruginosa against a novel lytic phage JJ01 exhibits hypersensitivity to colistin and reduces biofilm production","volume":"13","author":"Wannasrichan","year":"2022","journal-title":"Front. Microbiol."},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/16.22.10925","article-title":"Rapid isolation of \u03bb phage DNA in micro- and macro-variants","volume":"16","author":"Zabarovsky","year":"1988","journal-title":"Nucleic Acids Res."},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00345-22","article-title":"Phage resistance accompanies reduced fitness of uropathogenic escherichia coli in the urinary environment","volume":"7","author":"Zulk","year":"2022","journal-title":"mSphere"}],"updated-by":[{"DOI":"10.3389\/fcimb.2024.1391783","type":"erratum","label":"Erratum","source":"publisher","updated":{"date-parts":[[2024,3,6]],"date-time":"2024-03-06T00:00:00Z","timestamp":1709683200000}}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1280265\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,3,6]],"date-time":"2024-03-06T10:44:34Z","timestamp":1709721874000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1280265\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,17]]},"references-count":41,"alternative-id":["10.3389\/fcimb.2023.1280265"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2023.1280265","relation":{"erratum":[{"id-type":"doi","id":"10.3389\/fcimb.2024.1391783","asserted-by":"object"}]},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,17]]},"article-number":"1280265"}}