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This pathogen can acquire various antibiotic resistance genes, including those conferring resistance to last-resort antibiotics such as carbapenems. MDR <jats:italic>A. baumannii<\/jats:italic> is known to cause several infections, including pneumonia and urinary tract infections. Consequently, there is an urgent need to explore alternative therapies, and bacteriophage (phage) has emerged as a promising therapeutic approach for combating multidrug-resistant (MDR) infections.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Materials and methods<\/jats:title>\n            <jats:p>This study investigates the therapeutic potential of specific bacteriophages against MDR, particularly carbapenem-resistant <jats:italic>A. baumannii<\/jats:italic>, and evaluates lytic activity against 41 clinical isolates of MDR <jats:italic>A. baumannii<\/jats:italic>. The phages morphotypes were identified by transmission electron microscope. The stability of these phages was assessed under different conditions, including pH (2, 3, 4, 7, and 10\u201312), temperature (-80, -20, 4, 37, 50, 60, 70, and 80 <jats:sup>o<\/jats:sup>C), UV exposure (15, 30, 45, 60. 75, 90). Their antibacterial activity was also evaluated using a time-killing assay. Bacteriophage Insensitive Mutants (BIM) was assessed by MOI of 100. Genomic characterization was performed to predict protein-coding genes, life cycle, and suitability for therapeutic applications. Additionally, the safety and therapeutic efficacy of the phage were assessed using a cell viability MTT assay on adenocarcinomic human alveolar basal epithelial (A549) cells to evaluate the ability to rescue the lung cells from infection.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>Two phages, vB_AbaP_ZC2 (\u03a6ZC2) and vB_AbaM_ZC3 (\u03a6ZC3), were isolated from hospital wastewater in Egypt. The phages demonstrated lytic activity against 24.3% (<jats:italic>n<\/jats:italic>\u2009=\u200910) and 31.7% (<jats:italic>n<\/jats:italic>\u2009=\u200913) of the isolates, respectively. Phage \u03a6ZC2 demonstrated high EOP values (0.75\u20131) against AB23 and AB26, moderate activity on AB34 and AB35 (EOP\u2009=\u20090.19), and low or no activity on AB10, AB24, and AB31. Similarly, phage \u03a6ZC3 exhibited high EOP on AB24 (EOP\u2009=\u20091), moderate levels on AB12, AB29, and AB38, while showing low or no efficacy against the remaining tested isolates. The morphotypes of \u03a6ZC2 and \u03a6ZC3 are podovirus and myovirus, respectively. The two phages were amplified using a bioreactor and reached titers of approximately 10\u00b9\u2070 PFU\/ml in 2\u00a0L.\u03a6ZC2 was stable at a pH range from 3 to 12 approximately 10<jats:sup>8<\/jats:sup> PFU\/ml, while \u03a6ZC3 was stable at a pH range from 3 to 11 approximately 10<jats:sup>9<\/jats:sup> PFU\/ml compared to pH 7. \u03a6ZC2 was stable at -80, 37, and 50\u00a0\u00b0C approximately 10<jats:sup>8<\/jats:sup> PFU\/ml, while \u03a6ZC3 was stable at -80, 37,50, 60, and 70\u00a0\u00b0C with approximately 10<jats:sup>9<\/jats:sup> PFU\/ml compared to 4\u00a0\u00b0C. Additionally, the \u03a6ZC2 phage exhibited stability at 90\u00a0min, while \u03a6ZC3 phage exhibited stability at 75\u00a0min of exposure to UV light. The optimum MOI at which the \u03a6ZC2 and \u03a6ZC3 significantly reduced bacterial growth 0.1 and 0.01, respectively. The BIM frequency was higher for phage \u03a6ZC3 compared to \u03a6ZC2, indicating a slightly greater emergence of phage-resistant mutants with \u03a6ZC3. Whole genome sequencing and annotation did not identify markers for lysogeny or antibiotic resistance. Phylogenetic analysis classified \u03a6ZC2 and \u03a6ZC3 within the genera of <jats:italic>Obolenskvirus<\/jats:italic> and <jats:italic>Friunavirus<\/jats:italic>, respectively. \u03a6ZC3 was selected for its broad host range to be evaluated for rescuing A549 cells from MDR <jats:italic>A. baumannii<\/jats:italic> infection. \u03a6ZC3 phage was not cytotoxic to A549 cells and rescued lung cells cocultured, reducing the concentration of bacteria by approximately 5 logs with different MOIs, after 6\u00a0h of incubation.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>In this study, the two lytic phages have antibacterial activity against MDR <jats:italic>A. baumannii<\/jats:italic>. particularly, \u03a6ZC3 can be a potential therapy for pulmonary infections.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/s12985-025-02885-6","type":"journal-article","created":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T03:19:33Z","timestamp":1753845573000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Isolation and characterization of phages \u03a6ZC2 and \u03a6ZC3 against carbapenem-resistant Acinetobacter baumannii, and efficacy of \u03a6ZC3 on A549 cells"],"prefix":"10.1186","volume":"22","author":[{"given":"Kareem","family":"Essam","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Azza G.","family":"Kamel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bishoy Maher","family":"Zaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed","family":"Elhadidy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amal Ahmed Abdel","family":"Aziz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aysam","family":"Fayed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tamer","family":"Roshdy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayman","family":"El-Shibiny","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,7,30]]},"reference":[{"issue":"MAR","key":"2885_CR1","doi-asserted-by":"publisher","first-page":"249706","DOI":"10.3389\/FCIMB.2017.00055\/BIBTEX","volume":"7","author":"CR Lee","year":"2017","unstructured":"Lee CR, Lee JH, Park M, et al. 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