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This study characterizes Pseudomonas phage Banzai, a newly isolated Pbunavirus (family Lindbergviridae) with lytic activity against multiple P. aeruginosa isolates, including multidrug-resistant strains. Genomic analysis revealed a 66,189 bp genome, lacking antibiotic resistance or virulence factors, and suggested a headful packaging mechanism and the presence of a bidirectional component in the replication. In vivo experiments using Galleria mellonella showed therapeutic potential, significantly improving larval survival (87% at 24 h). Host range analysis revealed activity against 13 of 30 P. aeruginosa isolates, including members of O1, O3, O5 and O6 in silico predicted serogroups. Phylogenomic analyses place phage Banzai within the genus Pbunavirus, sharing 94.8% intergenomic similarity with its closest relatives, supporting its classification as a novel species. These findings highlight phage Banzai as a potential candidate for phage therapy, demonstrating genomic stability, a strictly lytic lifestyle, and in vivo efficacy.<\/jats:p>","DOI":"10.3390\/v17081088","type":"journal-article","created":{"date-parts":[[2025,8,6]],"date-time":"2025-08-06T13:25:11Z","timestamp":1754486711000},"page":"1088","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Pseudomonas Phage Banzai: Genomic and Functional Analysis of Novel Pbunavirus with Lytic Activity Against Pseudomonas aeruginosa"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1377-7153","authenticated-orcid":false,"given":"Andrei V.","family":"Chaplin","sequence":"first","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nina N.","family":"Sykilinda","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str. 16\/10, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-2136-4486","authenticated-orcid":false,"given":"George A.","family":"Skvortsov","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-5018-1265","authenticated-orcid":false,"given":"Konstantin S.","family":"Troshin","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-0362-0086","authenticated-orcid":false,"given":"Anna A.","family":"Vasilyeva","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sofia A.","family":"Shuraleva","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Artem A.","family":"Malkov","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladislav S.","family":"Simonov","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-8879-6516","authenticated-orcid":false,"given":"Boris A.","family":"Efimov","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lyudmila I.","family":"Kafarskaia","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4468-4091","authenticated-orcid":false,"given":"Konstantin A.","family":"Miroshnikov","sequence":"additional","affiliation":[{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str. 16\/10, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-5168-0927","authenticated-orcid":false,"given":"Anna A.","family":"Kuznetsova","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1646-9802","authenticated-orcid":false,"given":"Peter V.","family":"Evseev","sequence":"additional","affiliation":[{"name":"Pirogov Russian National Research Medical University, Ostrovityanova 1, 117997 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,6]]},"reference":[{"key":"ref_1","unstructured":"Naureen, Z., Dautaj, A., Anpilogov, K., Camilleri, G., Dhuli, K., Tanzi, B., Maltese, P.E., Cristofoli, F., De Antoni, L., and Beccari, T. 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