Abstract
Yersinia pestis, the causative agent of plague, is a highly virulent bacterium that poses a significant threat to human health. Preserving this bacterium in a viable state is crucial for research and diagnostic purposes. This paper presents and evaluates a simple lyophilization protocol for the long-term storage of Y. pestis strains from Fiocruz-CYP, aiming to explore its impact on viability and long-term stability, while replacing the currently used methodologies. The lyophilization tests were conducted using the non-virulent Y. pestis strain EV76, subjected to the lyophilization process under vacuum conditions. Viability assessment was performed to evaluate the effects of lyophilization and storage conditions on Y. pestis under multiple temperature conditions (− 80 °C, − 20 °C, 4–8 °C and room temperature). The lyophilization protocol employed in this study consistently demonstrated its efficacy in maintaining high viability rates for Y. pestis samples in a up to one year follow-up. The storage temperature that consistently exhibited the highest recovery rates was − 80 °C, followed by − 20 °C and 4–8 °C. Microscopic analysis of the post-lyophilized cultures revealed preserved morphological features, consistent with viable bacteria. The high viability rates observed in the preserved samples indicate the successful preservation of Y. pestis using this protocol. Overall, the presented lyophilization protocol provides a valuable tool for the long-term storage of Y. pestis, offering stability, viability, and functionality. By refining the currently used methods of lyophilization, this protocol can improve long-term preservation for Y. pestis strains collections, facilitating research efforts, diagnostic procedures, and the development of preventive and therapeutic strategies against plague.
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Acknowledgements
We are thankful to the Brazilian National Plague Reference Service staff for providing the samples to this study and to the Aggeu Magalhães Institute (FIOCRUZ-PE) for all furtherance.
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This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq grant #400734/2019–2) and by the Inova Novos Talentos—Fiocruz/Fundação Oswaldo Cruz (grant #VPPCB-008-FIO-18-2-73).
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All authors contributed to the study conception and design, as follows: I.V.R.: Conceptualization, Data curation, Formal analysis, Methodology, Writing (original draft). M.F.B.: Conceptualization, Data curation, Formal analysis, Methodology, Writing (original draft). M.S.: Project administration, Methodology. N.C.L.: Project administration. A.M.P.A.: Conceptualization, Funding acquisition, Project administration, Writing (original draft). All authors read and approved the final manuscript.
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Rocha, I.V., Bezerra, M.F., Sobreira, M. et al. Lyophilization for bacteria preservation: a promising approach for Yersinia pestis strains from an unique collection in Brazil (Fiocruz-CYP). Antonie van Leeuwenhoek 117, 61 (2024). https://doi.org/10.1007/s10482-024-01949-x
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DOI: https://doi.org/10.1007/s10482-024-01949-x