Optimization of the large-scale production for Erwinia amylovora bacteriophages.

BACKGROUND

Fire blight, caused by Erwinia amylovora, poses a significant threat to global agriculture, with antibiotic-resistant strains necessitating alternative solutions such as phage therapy. Scaling phage therapy to an industrial level requires efficient mass-production methods, particularly in optimizing the seed culture process. In this study, we investigated large-scale E. amylovora phage production by optimizing media supplementation and fermenter conditions, focusing on minimizing seed phages and pathogenic strains to reduce risks and improve the seed culture process.

RESULTS

We optimized the phage inoculum concentrations and media supplements to achieve higher phage yields comparable to or exceeding conventional methods. Laboratory-scale validation and refinement for fermenter-scale production allowed us to reduce bacterial and phage inoculum levels to 10⁵ CFU/mL and 10³ PFU/mL, respectively. Using fructose and sucrose supplements, the yields were comparable to conventional methods that use 10⁸ CFU/mL host bacteria and 10⁷ PFU/mL phages. Further pH adjustments in the fermenter increased yields by 16-303% across all phages tested.

CONCLUSIONS

We demonstrated the successful optimization and scale-up of E. amylovora phage production, emphasizing the potential for industrial bioprocessing with the reduced use of host cells and phage seeds. Overall, by refining key production parameters, we established a robust and scalable method for enhancing phage production efficiency.