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Assessing CaDPA levels, metabolic activity, and spore detection through deuterium labeling
Umeå University, Faculty of Science and Technology, Department of Physics. Swedish Defence Research Agency (FOI), Umeå, Sweden.ORCID iD: 0000-0002-0168-0197
Umeå University, Faculty of Science and Technology, Department of Physics.ORCID iD: 0009-0003-0399-9569
Umeå University, Faculty of Science and Technology, Department of Chemistry.ORCID iD: 0000-0002-3804-6421
Umeå University, Faculty of Science and Technology, Department of Physics. (The Biophysics and Biophotonics group)ORCID iD: 0000-0002-9835-3263
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2024 (English)In: The Analyst, ISSN 0003-2654, E-ISSN 1364-5528, Vol. 149, no 6, p. 1861-1871Article in journal (Refereed) Published
Abstract [en]

Many strains among spore-forming bacteria species are associated with food spoilage, foodborne disease, and hospital-acquired infections. Understanding the impact of environmental conditions and decontamination techniques on the metabolic activity, viability, and biomarkers of these spores is crucial for combatting them. To distinguish and track spores and to understand metabolic mechanisms, spores must be labeled. Staining or genetic modification are current methods for this, however, these methods can be time-consuming, and affect the viability and function of spore samples. In this work, we investigate the use of heavy water for permanent isotope labeling of spores and Raman spectroscopy for tracking sporulation/germination mechanisms. We also discuss the potential of this method in observing decontamination. We find that steady-state deuterium levels in the spore are achieved after only ∼48 h of incubation with 30% D2O-infused broth and sporulation, generating Raman peaks at cell silent region of 2200 and 2300 cm−1. These deuterium levels then decrease rapidly upon spore germination in non-deuterated media. We further find that unlike live spores, spores inactivated using various methods do not lose these Raman peaks upon incubation in growth media, suggesting these peaks may be used to indicate the viability of a spore sample. We further observe several Raman peaks exclusive to deuterated DPA, a spore-specific chemical biomarker, at e.g. 988 and 2300 cm−1, which can be used to track underlying changes in spores involving DPA. In conclusion, permanent spore labeling using deuterium offers a robust and non-invasive way of labeling bacterial spores for marking, viability determination, and characterising spore activity.

Place, publisher, year, edition, pages
Royal Society of Chemistry, 2024. Vol. 149, no 6, p. 1861-1871
National Category
Other Physics Topics Analytical Chemistry Other Biological Topics
Identifiers
URN: urn:nbn:se:umu:diva-221377DOI: 10.1039/d3an02162aISI: 001160646800001PubMedID: 38348676Scopus ID: 2-s2.0-85185190708OAI: oai:DiVA.org:umu-221377DiVA, id: diva2:1839729
Funder
Swedish Research Council, 2019-04016The Kempe Foundations, JCK-1916.2Swedish Armed Forces, 470-A400823Available from: 2024-02-21 Created: 2024-02-21 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, RasmusSil, Timir BaranOhlin, C. AndréAndersson, MagnusMalyshev, Dmitry

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