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However, development of phage resistance has led to unsuccessful therapeutic outcomes. In the current study, we applied phage training to minimize bacterial phage resistance and to improve treatment outcome by adapting the phage to their target hosts during co-evolution. We isolated and characterized a novel Pseudomonas aeruginosa N4-like lytic phage (PWJ) from wastewater in Yangzhou, China. PWJ is a double-stranded DNA podovirus that can efficiently lyse the model strain ATCC 27,853 and opportunistic pathogen PAO1. Genome sequencing of PWJ revealed features similar to those of the N4-like P. aeruginosa phage YH6. We used PWJ to screen for an evolved trained phage (WJ_Ev14) that restored infectivity to PWJ phage bacterial resisters. BLASTN analysis revealed that WJ_Ev14 is identical to its ancestor PWJ except for the amino acid substitution R1051S in its tail fiber protein. Moreover, phage adsorption tests and transmission electron microscopy of resistant bacteria demonstrated that the R1051S substitution was most likely the reason WJ_Ev14 could re-adsorb and regain infectivity. Furthermore, phage therapy assays in vitro and in a mouse P. aeruginosa lung infection model demonstrated that PWJ treatment resulted in improved clinical results and a reduction in lung bacterial load whereas the joint phage cocktail (PWJ+ WJ_Ev14) was better able to delay the emergence of resister bacteria. The phage cocktail (PWJ +WJ_Ev14) represents a promising candidate for inclusion in phage cocktails developed for clinical applications.<\/jats:p>","DOI":"10.1093\/ve\/vead067","type":"journal-article","created":{"date-parts":[[2023,12,11]],"date-time":"2023-12-11T20:43:47Z","timestamp":1702327427000},"source":"Crossref","is-referenced-by-count":12,"title":["Coevolutionary phage training and\u00a0Joint application delays the emergence of\u00a0phage resistance in\u00a0<i>Pseudomonas aeruginosa<\/i>"],"prefix":"10.1093","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0593-2475","authenticated-orcid":false,"given":"Mianzhi","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyi","family":"Wei","sequence":"additional","affiliation":[{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University , Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]},{"name":"Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) , College of Veterinary Medicine, Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Jiang","sequence":"additional","affiliation":[{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University , Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]},{"name":"Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) , College of Veterinary Medicine, Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Jiang","sequence":"additional","affiliation":[{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University , Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]},{"name":"Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) , College of Veterinary Medicine, Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junxuan","family":"Zhang","sequence":"additional","affiliation":[{"name":"National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, College of Veterinary Medicine, South China Agricultural University , Wushan Rd 483, Guangzhou, Guangdong 510642,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaolu","family":"He","sequence":"additional","affiliation":[{"name":"National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, College of Veterinary Medicine, South China Agricultural University , Wushan Rd 483, Guangzhou, Guangdong 510642,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiwen","family":"Ren","sequence":"additional","affiliation":[{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University , Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]},{"name":"Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) , College of Veterinary Medicine, Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zixuan","family":"Wang","sequence":"additional","affiliation":[{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University , Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]},{"name":"Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) , College of Veterinary Medicine, Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongxue","family":"Sun","sequence":"additional","affiliation":[{"name":"National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, College of Veterinary Medicine, South China Agricultural University , Wushan Rd 483, Guangzhou, Guangdong 510642,","place":["China"]},{"name":"South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture , Wushan Rd 483, Guangzhou, Guangdong 510642,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiqiang","family":"Wang","sequence":"additional","affiliation":[{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University , Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]},{"name":"Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) , College of Veterinary Medicine, Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]},{"name":"International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China , Daxue Rd 888, Yangzhou, Jiangsu 225009,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,11,18]]},"reference":[{"key":"2025071105263056600_R1","doi-asserted-by":"crossref","DOI":"10.3390\/ph11020060","article-title":"Parallel Evolution of\u00a0Host-Attachment Proteins in\u00a0Phage PP01 Populations Adapting to Escherichia Coli O157:H7","volume":"11","author":"Akusobi","year":"2018","journal-title":"Pharmaceuticals"},{"key":"2025071105263056600_R2","first-page":"e00066","article-title":"Phage Therapy in\u00a0the Postantibiotic 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