Comparative analysis of different preservation techniques for the storage of Staphylococcus phages aimed for the industrial development of phage-based antimicrobial products

Bacteriophages have been proven as effective antimicrobial agents in the treatment of infectious diseases and in other biocontrol applications including food preservation and disinfection. The extensive use of bacteriophages requires improved methodologies for medium- and long-term storage as well a...

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Bibliographic Details
Published in:PloS one 2018-10, Vol.13 (10), p.e0205728-e0205728
Main Authors: González-Menéndez, Eva, Fernández, Lucía, Gutiérrez, Diana, Rodríguez, Ana, Martínez, Beatriz, García, Pilar
Format: Article
Language:English
Subjects:
Additives
Alginates
Alginic acid
Antiinfectives and antibacterials
Antimicrobial agents
Bacteria
Bacterial infections
Bacteriophages
Biofilms
Biological control
Biology and Life Sciences
Comparative analysis
Diabetes
Disaccharides
Disinfection
Encapsulation
Food
Food preservation
Freeze drying
Glycerol
Industrial development
Infectious diseases
Medical treatment
Medicine and Health Sciences
Milk
Phages
Physical Sciences
Planning
Powder
Preservation
Research and Analysis Methods
Shipping
Skim milk
Sorbitol
Stabilizers (agents)
Staphylococcus
Staphylococcus infections
Storage
Studies
Trehalose
Viability
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Summary:Bacteriophages have been proven as effective antimicrobial agents in the treatment of infectious diseases and in other biocontrol applications including food preservation and disinfection. The extensive use of bacteriophages requires improved methodologies for medium- and long-term storage as well as for easy shipping. To this aim, we have determined the stability of four Staphylococcus phages (phiIPLA88, phiIPLA35, phiIPLA-RODI and phiIPLA-C1C) with antimicrobial potential at different temperatures (20°C/25°C, 4°C, -20°C, -80°C, -196°C) and during lyophilization (freeze drying) using several stabilizing additives (disaccharides, glycerol, sorbitol and skim milk). Differences between phages were observed at different temperatures (20°C/25°C, 4°C and -20°C), where phages were less stable. At lower temperatures (-80°C and -196°C), all phages showed good viability after 24 months regardless of the stabilizer. Differences between phages were also observed after lyophilization although the addition of skim milk yielded a dry powder with a stable titer after 24 months. As an alternative to facilitate storage and transportation, phage encapsulation has been also explored. Phage phiIPLA-RODI encapsulated in alginate capsules retained high viability when stored at 4°C for 6 months and at 20°C for 1 month. Moreover, the spray-dryer technique allowed obtaining dry powders containing viable encapsulated phages (phiIPLA-RODI and phiIPLA88) in both skim milk and trehalose for 12 months at 4°C. Storage of phages at 20°C was less effective; in fact, phiIPLA88 was stable for at least 12 months in trehalose but not in skim milk, while phiIPLA-RODI was stable only for 6 months in either stabilizer. These results suggest that encapsulated phages might be a suitable way for shipping phages.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0205728