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Real-time monitoring of H₂O₂ sterilization on individual bacillus atrophaeus spores by optical sensing with trapping Raman spectroscopy

  • Hydrogen peroxide (H₂O₂), a strong oxidizer, is a commonly used sterilization agent employed during aseptic food processing and medical applications. To assess the sterilization efficiency with H₂O₂, bacterial spores are common microbial systems due to their remarkable robustness against a wide variety of decontamination strategies. Despite their widespread use, there is, however, only little information about the detailed time-resolved mechanism underlying the oxidative spore death by H₂O₂. In this work, we investigate chemical and morphological changes of individual Bacillus atrophaeus spores undergoing oxidative damage using optical sensing with trapping Raman microscopy in real-time. The time-resolved experiments reveal that spore death involves two distinct phases: (i) an initial phase dominated by the fast release of dipicolinic acid (DPA), a major spore biomarker, which indicates the rupture of the spore’s core; and (ii) the oxidation of the remaining spore material resulting in the subsequent fragmentation of the spores’ coat. Simultaneous observation of the spore morphology by optical microscopy corroborates these mechanisms. The dependence of the onset of DPA release and the time constant of spore fragmentation on H₂O₂ shows that the formation of reactive oxygen species from H₂O₂ is the rate-limiting factor of oxidative spore death.

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Author:Morten Bertz, Denise MolinnusORCiD, Michael Josef SchöningORCiD, Takayuki Homma
Parent Title (English):Chemosensors
Place of publication:Basel
Document Type:Article
Year of Completion:2023
Date of first Publication:2023/08/10
Date of the Publication (Server):2023/09/21
Tag:Bacillus atrophaeus spores; DPA (dipicolinic acid); Raman spectroscopy; optical sensor setup; optical trapping; sterilization
First Page:Artikel 445
This article belongs to the Special Issue "Biosensors and Chemical Sensors for Food and Healthcare Monitoring—Celebrating the 10th Anniversary"
Institutes:FH Aachen / INB - Institut für Nano- und Biotechnologien
collections:Verlag / MDPI
Licence (German):License LogoCreative Commons - Namensnennung