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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: 2025-09-30Bibliographically approved
In thesis
1. Spotlight the killer: detecting harmful chemical and biological agents using optical spectroscopy
Open this publication in new window or tab >>Spotlight the killer: detecting harmful chemical and biological agents using optical spectroscopy
2025 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Lyset på mördaren : detektion av skadliga kemiska och biologiska ämnen med hjälp av optisk spektroskopi
Abstract [en]

Harmful chemical and biological agents are a significant threat to health and prosperity worldwide. Recent years have seen an increase in wars and conflicts around the globe, raising concerns about the potential deployment of chemical and biological warfare agents. On a less speculative level, harmful chemicals such as narcotic substances cause immense humanitarian and economic damage through overdoses and associated healthcare costs, while microbes such as pathogenic bacteria and parasites cause hospital-acquired infections and food spoilage at a cost of approximately 1 trillion euros every year. To combat the threat of these harmful agents, we must thus develop rapid and effective detection and diagnostic methods for harmful agents, allowing us to effectively deploy specific treatments and preventative measures.

Classically, while there exist numerous methods for the detection of both harmful chemical and biological agents, they often come with limitations that inhibit their effectiveness. These inhibitions often take the form of bulky equipment that is difficult to apply in the field or time-consuming preparation and measurement processes.

In this thesis we will explore an alternative category of assays for detecting and characterizing harmful materials – optical spectroscopy. Optical spectroscopy is a category of material characterization methods that use light to probe a material. While probing the material, we receive a signal characteristic of the molecules, chemical, and biological structure of our material. These optical spectroscopic methods, such as Raman spectroscopy and fluorescence spectroscopy, can be used to characterize a material within the span of minutes or even seconds, making them ideal for detection applications. Furthermore, they can often be made portable or even handheld, making them a great tool for initial field indication of harmful materials, ahead of thorough lab analysis.

I sincerely hope the studies presented herein can serve as a stepping stone to future technologies and detection assays, capable of saving both money and lives. 

Place, publisher, year, edition, pages
Umeå: Umeå University, 2025. p. 72
Keywords
Sensing, Raman spectroscopy, SERS, Fluorescence spectroscopy, CWA, nerve agents, bacterial spores, Cryptosporidium
National Category
Atom and Molecular Physics and Optics
Identifiers
urn:nbn:se:umu:diva-244830 (URN)978-91-8070-780-0 (ISBN)978-91-8070-779-4 (ISBN)
Public defence
2025-10-24, Aula Anatomica, Biologihuset, 907 36, Umeå, Umeå, 13:00 (English)
Opponent
Supervisors
Note

This work was done in collaboration with, and with support from, the Swedish Defece Research Agency (FOI).

Available from: 2025-10-03 Created: 2025-09-30 Last updated: 2025-10-22Bibliographically approved

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Öberg, RasmusDahlberg, TobiasMalyshev, DmitryAndersson, Magnus

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