Issue 9, 2023

Monitoring bacterial spore metabolic activity using heavy water-induced Raman peak evolution

Abstract

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 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.

Graphical abstract: Monitoring bacterial spore metabolic activity using heavy water-induced Raman peak evolution

Supplementary files

Article information

Article type
Paper
Submitted
16 Dec 2022
Accepted
04 Apr 2023
First published
04 Apr 2023
This article is Open Access
Creative Commons BY license

Analyst, 2023,148, 2141-2148

Monitoring bacterial spore metabolic activity using heavy water-induced Raman peak evolution

R. Öberg, T. Dahlberg, D. Malyshev and M. Andersson, Analyst, 2023, 148, 2141 DOI: 10.1039/D2AN02047E

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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