Development of Bacteriophage Therapy for Treatment of A. baumannii Infected Wounds

Panacea Pharmaceuticals proposes to develop an antibiotic product consisting of lytic bacteriophage that kills Acinetobacter baumannii for eventual use in human wound infections. The Phase 1 project will identify candidate bacteriophages from raw sewage and soil by mixing sewage/soil concentrates with A. baumannii, growing the bacteria on agar plates and isolating the resulting bacteriophage plaques. The isolated bacteriophage will be amplified by preparation of lysates on agar plates, then in liquid culture, followed by purification by either cesium chloride density gradient centrifugation or anion exchange column chromatography. The purified bacteriophage will be characterized by restriction endonuclease cleavage patterns on agarose gels, morphology by electron microscopy, estimation of latent period and burst size by plaque assay, virulence by lytic spectrum assay and plaque assay, host specificity for A. baumannii strains and related organisms, DNA sequencing of the bacteriophage genome and phylogenetic analysis by serological relationship, nucleic acid homology and morphological characteristics. Various dry and liquid storage conditions will be assessed for optimal stability of the bacteriophage. Safety and efficacy of the bacteriophage will be assessed by DNA sequence analysis for known toxin genes and by testing in two animal wound models, the burn and induced abscess models.