Development of a lyophilization process for Campylobacter bacteriophage storage and transport

Liang, Lu; Carrigy, Nicholas B.; Kariuki, Samuel; Muturi, Peter; Onsare, Robert; Nagel, Tobi; Vehring, Reinhard; Connerton, Phillippa L.; Connerton, Ian F.

Abstract

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. Bacteriophages are a sustainable alternative to control pathogenic bacteria in the post-antibiotic era. Despite promising reports, there are still obstacles to phage use, notably titer stability and transport‐associated expenses for applications in food and agriculture. In this study, we have developed a lyophilization approach to maintain phage titers, ensure efficacy and reduce transport costs of Campylobacter bacteriophages. Lyophilization methods were adopted with various excipients to enhance stabilization in combination with packaging options for international transport. Lyophilization of Eucampyvirinae CP30A using tryptone formed a cake that limited processing titer reduction to 0.35 ± 0.09 log10 PFU mL‐1. Transmission electron microscopy revealed the initial titer reduction was associated with capsid collapse of a subpopulation. Freeze‐dried phages were generally stable under refrigerated vacuum conditions and showed no significant titer changes over 3 months incubation at 4 °C (p = 0.29). Reduced stability was observed for lyophilized phages that were incubated either at 30 °C under vacuum or at 4 °C at 70% or 90% relative humidity. Refrigerated international transport and rehydration of the cake resulted in a total phage titer reduction of 0.81 ± 0.44 log10 PFU mL‐1. A significantly higher titer loss was observed for phages that were not refrigerated during transport (2.03 ± 0.32 log10 PFU mL‐1). We propose that lyophilization offers a convenient method to preserve and transport Campylobacter phages, with minimal titer reduction after the drying process.

Citation

Liang, L., Carrigy, N. B., Kariuki, S., Muturi, P., Onsare, R., Nagel, T., Vehring, R., Connerton, P. L., & Connerton, I. F. (2020). Development of a lyophilization process for Campylobacter bacteriophage storage and transport. Microorganisms, 8(2), 1-13. https://doi.org/10.3390/microorganisms8020282

• Journal Article Type: Article
• Acceptance Date: Feb 17, 2020
• Online Publication Date: Feb 19, 2020
• Publication Date: Feb 19, 2020
• Deposit Date: Feb 19, 2020
• Publicly Available Date: Feb 19, 2020
• Journal: Microorganisms
• Electronic ISSN: 2076-2607
• Publisher: MDPI
• Peer Reviewed: Peer Reviewed
• Volume: 8
• Issue: 2
• Article Number: 282
• Pages: 1-13
• DOI: https://doi.org/10.3390/microorganisms8020282
• Keywords: Campylobacter 2; bacteriophage 3; lyophilization 4; phage transportation 5
• Public URL: https://nottingham-repository.worktribe.com/output/3992861
• Publisher URL: https://www.mdpi.com/2076-2607/8/2/282