Antimicrobial resistance poses a growing threat to public health globally. Multidrug resistant Pseudomonas (P.) aeruginosa is detected in many infected wounds and is very challenging to treat with antibiotics. An alternative to antibiotics is to use bacteriophages, highly specific viruses able to kill even resistant bacteria. This work incorporates anti‐ P. aeruginosa Neko phages into monoaxial and coaxial electrospun fibers to explore their potential for treating infected wounds. Phages are blended with polyvinyl alcohol (PVA) solution and either processed directly into fibers or used as the core in coaxial electrospinning with polyvinylpyrrolidone (PVP) and PVP/ethyl cellulose (EC) shell solutions. Coaxial fibers stored at −20 °C show promising stability results, with negligible phage titer loss after 6 months of storage. Phage release can be controlled by varying the shell composition. Coaxial fibers with PVP as the shell (PVA/Su PVP + phage) demonstrate immediate release while fibers with PVP/EC as the shell (PVA/Su PVP/EC + phage) display extended‐release. The antibacterial efficacy of phage lysate and phage‐loaded fibers is studied by isothermal calorimetry and found to be unaffected by electrospinning. Thus, it appears that phage‐loaded electrospun fibers merit further investigation as potential wound dressing materials.

Pseudomonas aeruginosa is a major opportunistic pathogen with high levels of antibiotic resistance. Phage therapy represents a promising alternative for the treatment of difficult infections both alone and in combination with antibiotics. Here, we isolated and characterized three novel lytic myoviruses, Cisa, Nello, and Moonstruck. Genomic analysis revealed that Cisa and Nello belong to the Pbunavirus genus, while Moonstruck is a novel Pakpunavirus species. All lacked lysogeny, virulence, or resistance-associated genes, supporting their therapeutic suitability. Phage Nello and Moonstruck were active against P. aeruginosa Pa3GrPv, isolated from a patient with lung infection candidate for phage therapy. Moonstruck exhibited superior lytic activity with ciprofloxacin sub-MIC value (0.125 µg/mL), achieving bacterial suppression for 48 h. However, to improve the lytic efficacy of the phages on the clinical isolate, phage adaptation via serial passage was investigated. The killing efficacy of Nello was enhanced, whereas Moonstruck showed a less consistent improvement, suggesting phage-specific differences in evolutionary dynamics. Sequencing of the evolved phages revealed point mutations in tail-associated genes, potentially linked to a better phage–host interaction. These results support the use of phage–antibiotic combinations and directed evolution as strategies to enhance phage efficacy against drug-resistant infections. Overall, these findings support the therapeutic potential of the newly isolated phages in treating P. aeruginosa lung infections.