Animal-free peptones support bacteriophage propagation for therapeutic use without altering bactericidal activity or genomic integrity

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ID: 320982
2026
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Abstract
AIMS: Bacteriophage (phage) propagation has traditionally relied on bacterial culture media containing animal-derived ingredients; however, safety concerns with animal-derived materials for production of phages for therapeutic use limit their acceptability. We compared animal-free and traditional media formulations, and evaluated effects on phage yield, bactericidal activity, and genomic characteristics, hypothesizing no significant differences would be observed. METHODS AND RESULTS: Phages targeting Pseudomonas aeruginosa (n = 8) and Staphylococcus aureus (n = 1) were propagated in solid and liquid media containing animal-free (AF) or animal-derived (LB) peptones. Kinetic assays were used to assess phage suppression of host bacterial growth. Whole genome sequencing of phages and their bacterial hosts propagated in AF or LB broth was used to observe genomic differences between media formulations. In a mock therapeutic phage screen, spot tests, efficiency of plating (EOP) and kinetic assays were performed against non-host bacterial targets. Animal-free peptone did not impact phage yield, with both AF and LB phage stocks growing to high titers (≥108 PFU mL-1). Kinetic assay results showed similar suppression indices for AF and LB-grown phages. Comparisons of phage and bacterial genome annotations showed no major differences arising from media formulation. Spot test, EOP, and kinetic assay results in phage susceptibility tests were similar between AF and LB phages. CONCLUSIONS: Findings suggest animal-free peptones do not alter phage yield, bactericidal activity, or genomic characteristics, supporting use of animal-free medium for medicinal phage manufacture. This is one of the first studies to systematically combine phenotypic and genomic assessment of phages and hosts across animal-free and traditional media.
Reference Key
openalex_W7168250715 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Daniel R Laucirica, Phoebe G Carr, Mitchell G Hedges, Andrew Vaitekenas, Zlatibor Velickovic, Stephen M Stick, Samuel T Montgomery, Anthony Kicic
Journal Journal of applied microbiology
Year 2026
DOI
10.1093/jambio/lxag174
URL
https://doi.org/10.1093/jambio/lxag174
Keywords Keywords not found

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