{"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":1790168654982,"version":"4.1.0"},"reference-count":43,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,7,5]],"date-time":"2025-07-05T00:00:00Z","timestamp":1751673600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"research grants from Shandong Provincial Natural Science Foundation","award":["ZR2022MC001"],"award-info":[{"award-number":["ZR2022MC001"]}]},{"name":"research grants from Shandong Provincial Natural Science Foundation","award":["25-1-5-xdny-30-nsh"],"award-info":[{"award-number":["25-1-5-xdny-30-nsh"]}]},{"name":"research grants from Shandong Provincial Natural Science Foundation","award":["24-1-8-xdny-4-nsh"],"award-info":[{"award-number":["24-1-8-xdny-4-nsh"]}]},{"name":"research grants from Shandong Provincial Natural Science Foundation","award":["24-1-8-xdny-21-nsh"],"award-info":[{"award-number":["24-1-8-xdny-21-nsh"]}]},{"name":"research grants from Shandong Provincial Natural Science Foundation","award":["TS20220701"],"award-info":[{"award-number":["TS20220701"]}]},{"name":"the Scientific and Technological Projects of Qingdao","award":["ZR2022MC001"],"award-info":[{"award-number":["ZR2022MC001"]}]},{"name":"the Scientific and Technological Projects of Qingdao","award":["25-1-5-xdny-30-nsh"],"award-info":[{"award-number":["25-1-5-xdny-30-nsh"]}]},{"name":"the Scientific and Technological Projects of Qingdao","award":["24-1-8-xdny-4-nsh"],"award-info":[{"award-number":["24-1-8-xdny-4-nsh"]}]},{"name":"the Scientific and Technological Projects of Qingdao","award":["24-1-8-xdny-21-nsh"],"award-info":[{"award-number":["24-1-8-xdny-21-nsh"]}]},{"name":"the Scientific and Technological Projects of Qingdao","award":["TS20220701"],"award-info":[{"award-number":["TS20220701"]}]},{"name":"Taishan Industrial Experts Program","award":["ZR2022MC001"],"award-info":[{"award-number":["ZR2022MC001"]}]},{"name":"Taishan Industrial Experts Program","award":["25-1-5-xdny-30-nsh"],"award-info":[{"award-number":["25-1-5-xdny-30-nsh"]}]},{"name":"Taishan Industrial Experts Program","award":["24-1-8-xdny-4-nsh"],"award-info":[{"award-number":["24-1-8-xdny-4-nsh"]}]},{"name":"Taishan Industrial Experts Program","award":["24-1-8-xdny-21-nsh"],"award-info":[{"award-number":["24-1-8-xdny-21-nsh"]}]},{"name":"Taishan Industrial Experts Program","award":["TS20220701"],"award-info":[{"award-number":["TS20220701"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The high pathogenicity rate of carbapenem-resistant Pseudomonas aeruginosa (CRPA) has resulted in substantial economic losses for humans and the breeding industry. Consequently, there is an urgent need to develop new alternatives to mitigate antibiotic use. Phage therapy has demonstrated promising results in numerous studies. In this study, lytic phages targeting CRPA were isolated from feces and river water samples in Shandong, China. A total of 94 phage strains with CRPA as hosts were obtained, exhibiting lysis rates that ranged from 29% to 76% for P. aeruginosa derived from humans and different types of animals (n = 246). We further examined five representative phages, the host bacteria of which were CRPA from clinical patients and poultry, and these phages included two myoviruses and three podoviruses. Their optimal multiplicities of infection (MOIs) ranged from 10\u22123 to 10\u22125, with latent periods of less than 5 to 15 min and burst durations of 140 to 175 min, resulting in burst sizes of 133 to 352 PFU\/cell. All five phages exhibited the ability to survive at temperatures up to 60 \u00b0C and within pH levels of 3 to 11. Whole-genome sequencing revealed that these five phages were all double-stranded DNA phages and did not possess resistance genes or virulence factors. The two myoviruses, sharing similar sequences, were classified into the genus Pakpunavirus, with a size of 92,509 bp and 92,293 bp, 149 to 152 ORFs and 20 to 22 tRNAs. In contrast, the three similar podoviruses belong to the genus Phikmvvirus and all contained a perforin\u2013lyase system, with a size of 43.35 kb, a GC content of 62%, 49 to 50 ORFs and 16 to 20 tRNAs. A spray disinfection experiment demonstrated that the phage cocktail exhibited a high sterilization effect after spraying and showed good efficacy against cement and metal surfaces. This study provides foundational information for further research into the elimination of CRPA in the environment.<\/jats:p>","DOI":"10.3390\/microorganisms13071587","type":"journal-article","created":{"date-parts":[[2025,7,7]],"date-time":"2025-07-07T02:53:07Z","timestamp":1751856787000},"page":"1587","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Characterization, Genomic Analysis and Application of Five Lytic Phages Against Carbapenem-Resistant Pseudomonas aeruginosa"],"prefix":"10.3390","volume":"13","author":[{"given":"Li-Ping","family":"Zhang","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang-An","family":"Li","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongda","family":"Zhao","sequence":"additional","affiliation":[{"name":"Qingdao Bolin Biotechnology Co., Ltd., Qingdao 266000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zeqing","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junjie","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng-Jing","family":"Song","sequence":"additional","affiliation":[{"name":"Institute of Plant Protection, Qingdao Academy of Agricultural Sciences, Qingdao 266100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1178-2493","authenticated-orcid":false,"given":"Bao-Tao","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Handley, J.A., Park, S.H., Kim, S.A., and Ricke, S.C. (2018). Microbiome Profiles of Commercial Broilers Through Evisceration and Immersion Chilling During Poultry Slaughter and the Identification of Potential Indicator Microorganisms. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.00345"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Gomila, A., Carratal\u00e0, J., Badia, J.M., Camprub\u00ed, D., Piriz, M., Shaw, E., Diaz-Brito, V., Espejo, E., Nicol\u00e1s, C., and Brugu\u00e9s, M. (2018). Preoperative oral antibiotic prophylaxis reduces Pseudomonas aeruginosa surgical site infections after elective colorectal surgery: A multicenter prospective cohort study. BMC Infect. Dis., 18.","DOI":"10.1186\/s12879-018-3413-1"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/j.drudis.2018.07.003","article-title":"Updates on the pathogenicity status of Pseudomonas aeruginosa","volume":"24","author":"Azam","year":"2019","journal-title":"Drug Discov. Today"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e71","DOI":"10.1590\/s1678-9946202163071","article-title":"Carbapenem-resistant Pseudomonas aeruginosa strains: A worrying health problem in intensive care units","volume":"63","author":"Souza","year":"2021","journal-title":"Rev. Inst. Med. Trop. Sao Paulo"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1016\/j.jmii.2021.07.015","article-title":"Multicenter surveillance of antimicrobial susceptibilities and resistance mechanisms among Enterobacterales species and non-fermenting Gram-negative bacteria from different infection sources in Taiwan from 2016 to 2018","volume":"55","author":"Jean","year":"2022","journal-title":"J. Microbiol. Immunol. Infect. = Wei Mian Yu Gan Ran Za Zhi"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/j.diagmicrobio.2017.11.003","article-title":"SPM-1-producing Pseudomonas aeruginosa ST277 clone recovered from microbiota of migratory birds","volume":"90","author":"Martins","year":"2018","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"114378","DOI":"10.1016\/j.addr.2022.114378","article-title":"Novel antimicrobial agents for combating antibiotic-resistant bacteria","volume":"187","author":"Chang","year":"2022","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1146\/annurev-med-080219-122208","article-title":"Phage Therapy for Antibiotic-Resistant Bacterial Infections","volume":"73","author":"Hatfull","year":"2022","journal-title":"Annu. Rev. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"veaa060","DOI":"10.1093\/ve\/veaa060","article-title":"Broadscale phage therapy is unlikely to select for widespread evolution of bacterial resistance to virus infection","volume":"6","author":"Cohan","year":"2020","journal-title":"Virus Evol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1038\/icb.1960.34","article-title":"Lysogeny in Pseudomonas aeruginosa","volume":"38","author":"Holloway","year":"1960","journal-title":"Aust. J. Exp. Biol. Med. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1016\/j.micinf.2012.11.002","article-title":"Efficacy of cocktail phage therapy in treating Vibrio cholerae infection in rabbit model","volume":"15","author":"Jaiswal","year":"2013","journal-title":"Microbes Infect."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5646","DOI":"10.1128\/AEM.00757-12","article-title":"Effects of sequential and simultaneous applications of bacteriophages on populations of Pseudomonas aeruginosa in vitro and in wax moth larvae","volume":"78","author":"Hall","year":"2012","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1111\/j.1749-4486.2009.01973.x","article-title":"A controlled clinical trial of a therapeutic bacteriophage preparation in chronic otitis due to antibiotic-resistant Pseudomonas aeruginosa; a preliminary report of efficacy","volume":"34","author":"Wright","year":"2009","journal-title":"Clin. Otolaryngol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Fong, S.A., Drilling, A., Morales, S., Cornet, M.E., Woodworth, B.A., Fokkens, W.J., Psaltis, A.J., Vreugde, S., and Wormald, P.J. (2017). Activity of Bacteriophages in Removing Biofilms of Pseudomonas aeruginosa Isolates from Chronic Rhinosinusitis Patients. Front. Cell. Infect. Microbiol., 7.","DOI":"10.3389\/fcimb.2017.00418"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5340","DOI":"10.1128\/AEM.01434-14","article-title":"Inhibition of biofilm formation by T7 bacteriophages producing Quorum-Quenching enzymes","volume":"80","author":"Pei","year":"2014","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.chom.2019.01.014","article-title":"Phage Therapy: A Renewed Approach to Combat Antibiotic-Resistant Bacteria","volume":"25","author":"Kortright","year":"2019","journal-title":"Cell Host Microbe"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.cell.2022.11.017","article-title":"Phage therapy: From biological mechanisms to future directions","volume":"186","author":"Strathdee","year":"2023","journal-title":"Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"522","DOI":"10.3947\/ic.2024.0097","article-title":"Effect of Antimicrobial Wipes on Hospital-Associated Bacterial and Fungal Strains","volume":"56","author":"Chun","year":"2024","journal-title":"Infect. Chemother."},{"key":"ref_19","first-page":"2155","article-title":"Pseudomonas aeruginosa infection of avian origin: Zoonosis and one health implications","volume":"14","year":"2021","journal-title":"Vet. World"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Li, Z., Li, W., Ma, W., Ding, Y., Zhang, Y., Yang, Q., Wang, J., and Wang, X. (2021). Characterization and Application of a Lytic Phage D10 against Multidrug-Resistant Salmonella. Viruses, 13.","DOI":"10.3390\/v13081626"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"110480","DOI":"10.1016\/j.foodres.2021.110480","article-title":"Isolation, characterization and comparison of lytic Epseptimavirus phages targeting Salmonella","volume":"147","author":"Cong","year":"2021","journal-title":"Food Res. Int."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/S0065-2164(10)70007-1","article-title":"Bacteriophage host range and bacterial resistance","volume":"70","author":"Hyman","year":"2010","journal-title":"Adv. Appl. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"491","DOI":"10.3923\/pjbs.2020.491.500","article-title":"Isolation and Characterization of Pseudomonas aeruginosa and its Virulent Bacteriophages","volume":"23","author":"Marei","year":"2020","journal-title":"Pak. J. Biol. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Shmidov, E., Lebenthal-Loinger, I., Roth, S., Karako-Lampert, S., Zander, I., Shoshani, S., Danielli, A., and Banin, E. (2022). PrrT\/A, a Pseudomonas aeruginosa Bacterial Encoded Toxin-Antitoxin System Involved in Prophage Regulation and Biofilm Formation. Microbiol. Spectr., 10.","DOI":"10.1128\/spectrum.01182-22"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"199178","DOI":"10.1016\/j.virusres.2023.199178","article-title":"Therapeutic effect and anti-biofilm ability assessment of a novel phage, phiPA1-3, against carbapenem-resistant Pseudomonas aeruginosa","volume":"335","author":"Tsai","year":"2023","journal-title":"Virus Res."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Sun, W.J., Liu, C.F., Yu, L., Cui, F.J., Zhou, Q., Yu, S.L., and Sun, L. (2012). A novel bacteriophage KSL-1 of 2-Keto-gluconic acid producer Pseudomonas fluorescens K1005: Isolation, characterization and its remedial action. BMC Microbiol., 12.","DOI":"10.1186\/1471-2180-12-127"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Wang, X., Tang, J., Dang, W., Xie, Z., Zhang, F., Hao, X., Sun, S., Liu, X., Luo, Y., and Li, M. (2023). Isolation and Characterization of Three Pseudomonas aeruginosa Viruses with Therapeutic Potential. Microbiol. Spectr., 11.","DOI":"10.1128\/spectrum.04636-22"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e2055","DOI":"10.7717\/peerj.2055","article-title":"Assessing Illumina technology for the high-throughput sequencing of bacteriophage genomes","volume":"4","author":"Rihtman","year":"2016","journal-title":"PeerJ"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Tang, Z., Tang, N., Wang, X., Ren, H., Zhang, C., Zou, L., Han, L., Guo, L., and Liu, W. (2023). Characterization of a lytic Escherichia coli phage CE1 and its potential use in therapy against avian pathogenic Escherichia coli infections. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1091442"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2173","DOI":"10.1007\/s11095-014-1617-7","article-title":"Quality and safety requirements for sustainable phage therapy products","volume":"32","author":"Blasdel","year":"2015","journal-title":"Pharm. Res."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Victor, K., Olga, S., Elena, P., Maria, B., Sergey, K., Alla, K., Elena, C., Leonid, K., Damian, M., and Olga, P. (2016). Modular Approach to Select Bacteriophages Targeting Pseudomonas aeruginosa for Their Application to Children Suffering with Cystic Fibrosis. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.01631"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6570","DOI":"10.3390\/v7122958","article-title":"What Can We Learn from a Metagenomic Analysis of a Georgian Bacteriophage Cocktail?","volume":"7","author":"Zschach","year":"2015","journal-title":"Viruses"},{"key":"ref_33","first-page":"8","article-title":"Isolation, characterization, and genomic analysis of vBPaePTUMSP121, a new lytic bacteriophage infecting Pseudomonas aeruginosa","volume":"168","author":"Haniyeh","year":"2022","journal-title":"Arch. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Khashayar, S., Mohadeseh, B., Lili, Z., Abolghasem, H., Tao, H., Hongduo, B., Mojtaba, M., Maoda, P., Heye, W., and Ruicheng, W. (2021). Biodiversity of New Lytic Bacteriophages Infecting Shigella spp. in Freshwater Environment. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.619323"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1613","DOI":"10.4014\/jmb.1208.08072","article-title":"Biocontrol potential of a lytic bacteriophage PE204 against bacterial wilt of tomato","volume":"22","author":"Bae","year":"2012","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Tan, C.W., Rukayadi, Y., Hasan, H., Abdul-Mutalib, N.A., Jambari, N.N., Hara, H., Thung, T.Y., Lee, E., and Radu, S. (2021). Isolation and Characterization of Six Vibrio parahaemolyticus Lytic Bacteriophages From Seafood Samples. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.616548"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Zamora, P.F., Reidy, T.G., Armbruster, C.R., Sun, M., Van Tyne, D., Turner, P.E., Koff, J.L., and Bomberger, J.M. (2024). Lytic bacteriophages induce the secretion of antiviral and proinflammatory cytokines from human respiratory epithelial cells. PLoS Biol., 22.","DOI":"10.1371\/journal.pbio.3002566"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Wang, X., Han, L., Rong, J., Ren, H., Liu, W., and Zhang, C. (2022). Endolysins of bacteriophage vB_Sal-S-S10 can naturally lyse Salmonella enteritidis. BMC Vet. Res., 18.","DOI":"10.1186\/s12917-022-03514-y"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Amgarten, D., Martins, L.F., Lombardi, K.C., Antunes, L.P., de Souza, A.P.S., Nicastro, G.G., Kitajima, E.W., Quaggio, R.B., Upton, C., and Setubal, J.C. (2017). Three novel Pseudomonas phages isolated from composting provide insights into the evolution and diversity of tailed phages. BMC Genom., 18.","DOI":"10.1186\/s12864-017-3729-z"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Chegini, Z., Khoshbayan, A., Taati Moghadam, M., Farahani, I., Jazireian, P., and Shariati, A. (2020). Bacteriophage therapy against Pseudomonas aeruginosa biofilms: A review. Ann. Clin. Microbiol. Antimicrob., 19.","DOI":"10.1186\/s12941-020-00389-5"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4239","DOI":"10.1038\/s41467-022-31837-9","article-title":"Personalized bacteriophage therapy to treat pandrug-resistant spinal Pseudomonas aeruginosa infection","volume":"13","author":"Ferry","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1352","DOI":"10.1016\/j.chom.2022.09.010","article-title":"Host happy hour: Phage cocktail targets IBD-associated microbes","volume":"30","author":"Mimee","year":"2022","journal-title":"Cell Host Microbe"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Kovacs, C.J., Rapp, E.M., Rankin, W.R., McKenzie, S.M., Brasko, B.K., Hebert, K.E., Bachert, B.A., Kick, A.R., Burpo, F.J., and Barnhill, J.C. (2024). Combinations of Bacteriophage Are Efficacious against Multidrug-Resistant Pseudomonas aeruginosa and Enhance Sensitivity to Carbapenem Antibiotics. Viruses, 16.","DOI":"10.3390\/v16071000"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/7\/1587\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:05:08Z","timestamp":1760033108000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/7\/1587"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,5]]},"references-count":43,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["microorganisms13071587"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms13071587","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,5]]}}}