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Monitoring bacterial spore metabolic activity using heavy water-induced Raman peak evolution
Umeå University, Faculty of Science and Technology, Department of Physics.ORCID iD: 0000-0002-0168-0197
Umeå University, Faculty of Science and Technology, Department of Physics.
Umeå University, Faculty of Science and Technology, Department of Physics.ORCID iD: 0000-0002-0496-6692
Umeå University, Faculty of Science and Technology, Department of Physics. Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR). (The Biophysics and Biophotonics group)ORCID iD: 0000-0002-9835-3263
2023 (English)In: The Analyst, ISSN 0003-2654, E-ISSN 1364-5528, Vol. 148, no 9, p. 2141-2148Article in journal (Refereed) Published
Abstract [en]

Endospore-forming bacteria are associated with food spoilage, food poisoning, and infection in hospitals. Therefore, methods to monitor spore metabolic activity and verify sterilization are of great interest. However, current methods for tracking metabolic activity are time-consuming and resource intensive. This work investigates isotope labeling and Raman microscopy as a low-cost rapid alternative. Specifically, we monitor the Raman spectrum of enterotoxic \textit{B. cereus} spores undergoing germination and cell division in D2O-infused broth. During germination and cell division, water is metabolized and deuterium from the broth is incorporated into proteins and lipids, resulting in the appearance of a Raman peak related to C-D bonds at 2190 cm-1. We find that a significant C-D peak appears after 2 h of incubation at 37◦C. Further, we found that the peak appearance coincides with the observed first cell division indicating little metabolic activity during germination. Lastly, the germination and cell growth rate of spores were not affected by adding 30 % heavy water to the broth. This shows the potential for real-time monitoring of metabolic activity from a bacterial spore to a dividing cell. In conclusion, our work proposes tracking the evolution of the C-D Raman peak in spores incubated with D2O-infused broth as an effective and time-, and cost-efficient method to monitor the outgrowth of a spore population, simultaneously allowing us to track for how long the bacteria have grown and divided.

Place, publisher, year, edition, pages
Royal Society of Chemistry, 2023. Vol. 148, no 9, p. 2141-2148
Keywords [en]
Bacterial spores, Heavy water, D2O, Raman spectroscopy, Viability, Germination
National Category
Other Physics Topics Analytical Chemistry Microbiology
Identifiers
URN: urn:nbn:se:umu:diva-206398DOI: 10.1039/D2AN02047EISI: 000968915700001PubMedID: 37040186Scopus ID: 2-s2.0-85153492235OAI: oai:DiVA.org:umu-206398DiVA, id: diva2:1748838
Funder
Swedish Research Council, 2019-04016The Kempe Foundations, JCK1916.2Swedish Armed Forces, 470-A400821Available from: 2023-04-04 Created: 2023-04-04 Last updated: 2023-06-09Bibliographically approved

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Öberg, RasmusDahlberg, TobiasMalyshev, DmitryAndersson, Magnus

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