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Use of Trehalose as an Additive to Bacteriophage Vb_Pd_PDCC-1: Long-Term Preservation Analysis and Its Biocontrol Against Vibrio diabolicus Infection

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Abstract

Phage therapy is a promising alternative to control bacterial diseases and the increasing problem of antibiotic resistance. In this sense, this research evaluates the viability of lyophilized vibrio phage vB_Pd_PDCC-1 using trehalose as a preservative excipient at different concentrations (4, 2, 1, and 0.5% w/v) and its potential for phage therapy application against a pathogenic bacteria Vibrio diabolicus in brine shrimp nauplii (Artemia franciscana). The lyophilized phages were stored at 4 and 23 °C and rehydrated using biological sterile saline solution to test their viability at days 1, 15, and 60 post-lyophilization. The results showed that trehalose is beneficial in maintaining the viability of post-lyophilization phages (without titer losses) at 4 °C and even at room temperature (23 °C). When lyophilized phages with 4% w/v trehalose concentration were stored at 23 °C, they had not titer losses among the trials; viability and titer concentration were maintained up to 60 days at log 7. The use of lyophilized phage PDCC-1 increased brine shrimp survival and reduced Vibrio concentrations. The present study has identified trehalose as a promising lyophilization excipient to effectively preserve lyophilized bacteriophages for biotechnological applications and long-term storage.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors acknowledge the technical assistance of Norma Angélica Ochoa Álvarez, José Manuel Melero Astorga, and María Sofía Ramos Galván at CIBNOR. Dr. Luis Daniel Moreno-Figueroa (CVU: 336817) is a recipient of a postdoctoral fellowship from the Consejo Nacional de Ciencia y Tecnología of Mexico (CONACYT) and Diana Fischer for English edition.

Funding

This work was funded by the Project “Infraestructura CONACYT” No. 316934 awarded to Dr. Luis Hernández-Adame.

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Authors and Affiliations

  1. Nanotechnology and Microbial Biocontrol Group, Centro de Investigaciones Biológicas del Noroeste, 23096, La Paz, Baja California Sur, Mexico

    Luis Daniel Moreno-Figueroa & Luis Hernández-Adame

  2. Centro de Investigaciones Biológicas del Noroeste S.C. (CIBNOR), Av. IPN 195, Col Playa Palo de Sta. Rita Sur, C.P. 23096, La Paz, B.C.S, Mexico

    Eduardo Quiroz-Guzmán & Dariel Tovar-Ramírez

  3. Laboratorio de Biopolímeros y Nanoestructuras, Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, C.P. 78210, San Luis Potosí, Mexico

    Gabriela Palestino & Cándida A. Cisneros-Covarrubias

  4. Cátedras CONACYT-Centro de Investigaciones Biológicas del Noroeste, 23096, La Paz, Baja California Sur, Mexico

    Luis Hernández-Adame

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Contributions

LDMF was contributed to conceptualization, data curation, formal analysis, methodology, and writing of the original draft. LHA contributed to conceptualization, data curation, formal analysis, funding acquisition, project administration, supervision, and writing, reviewing, and editing of the manuscript. GP contributed to data curation, validation, and formal analysis. CACC contributed to data curation, validation, and formal analysis. EQG contributed to data curation, validation, formal analysis, and writing, reviewing, and editing of the manuscript. DTR contributed to data curation, validation, formal analysis, and writing, reviewing, and editing of the manuscript.

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Correspondence to Luis Hernández-Adame.

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Moreno-Figueroa, L.D., Quiroz-Guzmán, E., Tovar-Ramírez, D. et al. Use of Trehalose as an Additive to Bacteriophage Vb_Pd_PDCC-1: Long-Term Preservation Analysis and Its Biocontrol Against Vibrio diabolicus Infection. Curr Microbiol 80, 372 (2023). https://doi.org/10.1007/s00284-023-03487-7

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