Autofluorescence properties of wound-associated bacteria cultured under various temperature, salinity, and pH conditions

Abstract Background Bacterial autofluorescence plays a vital role in photodiagnosis (PD) and antimicrobial photodynamic therapy (aPDT), yet the autofluorescence properties of wound-associated bacteria and their responses to the physicochemical microenvironment, remain underexplored. Here, we investi...

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Main Authors: Xiaofen Sun, Yikun Wang, Ao Du, Meili Dong, Yuhan Wang, Yuanzhi Zhang, Yang Zhang, Yao Huang, Xiang Huang, Yong Liu, Jingshu Ni
Format: Article
Language:English
Published: BMC 2025-08-01
Series:BMC Microbiology
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Online Access:https://doi.org/10.1186/s12866-025-04200-3
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author Xiaofen Sun
Yikun Wang
Ao Du
Meili Dong
Yuhan Wang
Yuanzhi Zhang
Yang Zhang
Yao Huang
Xiang Huang
Yong Liu
Jingshu Ni
author_facet Xiaofen Sun
Yikun Wang
Ao Du
Meili Dong
Yuhan Wang
Yuanzhi Zhang
Yang Zhang
Yao Huang
Xiang Huang
Yong Liu
Jingshu Ni
author_sort Xiaofen Sun
collection DOAJ
description Abstract Background Bacterial autofluorescence plays a vital role in photodiagnosis (PD) and antimicrobial photodynamic therapy (aPDT), yet the autofluorescence properties of wound-associated bacteria and their responses to the physicochemical microenvironment, remain underexplored. Here, we investigated the bacterial autofluorescence of Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus under various culture conditions, including different temperatures, NaCl concentrations, and pH levels found within wounds. Fluorescence imaging was employed to quantify red fluorescence intensity, while fluorescence spectrometry was used to correlate the observed fluorescence with spectral profiles, benchmarking them against coproporphyrin and protoporphyrin IX. Results Our results revealed that the selected bacteria emitted red fluorescence in vitro, consistent with their known porphyrin biosynthesis capabilities. The intensity of red fluorescence was primarily dependent on the bacterial species, growth phase, and culture conditions. Elevated culture temperature accelerated the fluorescence metabolism, whereas increasing NaCl concentrations and alkaline pH levels inhibited red fluorescence in a dose-dependent manner. Linear regression revealed a strong positive correlation between red fluorescence intensity and peak fluorescence emission when excited at 405 nm. The biosynthesis of endogenous porphyrins varied both across and within bacterial species, with distinct porphyrins produced under specific conditions. The emission spectra of the gram-positive S. aureus consistently aligned with coproporphyrin, while the gram-negative A. baumannii, E. coli, and K. pneumoniae typically displayed fluorescence peaks characteristic of protoporphyrin IX. Notably, K. pneumoniae shifted to coproporphyrin with extended culture duration or more favourable conditions, and E. coli exhibited a similar transition as the pH increased to 9. Conclusions We concluded that the local physicochemical conditions found within wounds could affect the autofluorescence and endogenous porphyrins of the bacteria, which may, in turn, have connotations for PD and aPDT.
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issn 1471-2180
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publishDate 2025-08-01
publisher BMC
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series BMC Microbiology
spelling doaj-art-4c9783f0bf1c41419bbe88634ffb1e3a2025-08-20T03:42:23ZengBMCBMC Microbiology1471-21802025-08-0125111710.1186/s12866-025-04200-3Autofluorescence properties of wound-associated bacteria cultured under various temperature, salinity, and pH conditionsXiaofen Sun0Yikun Wang1Ao Du2Meili Dong3Yuhan Wang4Yuanzhi Zhang5Yang Zhang6Yao Huang7Xiang Huang8Yong Liu9Jingshu Ni10Anhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesDepartment of Anesthesiology, Division of Life Sciences and Medicine, First Affiliated Hospital of USTC, University of Science and Technology of ChinaAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAnhui Institute of Optics and Fine Mechanics, Hefei Institutes of Physical Science, Chinese Academy of SciencesAbstract Background Bacterial autofluorescence plays a vital role in photodiagnosis (PD) and antimicrobial photodynamic therapy (aPDT), yet the autofluorescence properties of wound-associated bacteria and their responses to the physicochemical microenvironment, remain underexplored. Here, we investigated the bacterial autofluorescence of Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus under various culture conditions, including different temperatures, NaCl concentrations, and pH levels found within wounds. Fluorescence imaging was employed to quantify red fluorescence intensity, while fluorescence spectrometry was used to correlate the observed fluorescence with spectral profiles, benchmarking them against coproporphyrin and protoporphyrin IX. Results Our results revealed that the selected bacteria emitted red fluorescence in vitro, consistent with their known porphyrin biosynthesis capabilities. The intensity of red fluorescence was primarily dependent on the bacterial species, growth phase, and culture conditions. Elevated culture temperature accelerated the fluorescence metabolism, whereas increasing NaCl concentrations and alkaline pH levels inhibited red fluorescence in a dose-dependent manner. Linear regression revealed a strong positive correlation between red fluorescence intensity and peak fluorescence emission when excited at 405 nm. The biosynthesis of endogenous porphyrins varied both across and within bacterial species, with distinct porphyrins produced under specific conditions. The emission spectra of the gram-positive S. aureus consistently aligned with coproporphyrin, while the gram-negative A. baumannii, E. coli, and K. pneumoniae typically displayed fluorescence peaks characteristic of protoporphyrin IX. Notably, K. pneumoniae shifted to coproporphyrin with extended culture duration or more favourable conditions, and E. coli exhibited a similar transition as the pH increased to 9. Conclusions We concluded that the local physicochemical conditions found within wounds could affect the autofluorescence and endogenous porphyrins of the bacteria, which may, in turn, have connotations for PD and aPDT.https://doi.org/10.1186/s12866-025-04200-3Bacterial autofluorescenceEndogenous porphyrinsTemperatureNaCl concentrationpH
spellingShingle Xiaofen Sun
Yikun Wang
Ao Du
Meili Dong
Yuhan Wang
Yuanzhi Zhang
Yang Zhang
Yao Huang
Xiang Huang
Yong Liu
Jingshu Ni
Autofluorescence properties of wound-associated bacteria cultured under various temperature, salinity, and pH conditions
BMC Microbiology
Bacterial autofluorescence
Endogenous porphyrins
Temperature
NaCl concentration
pH
title Autofluorescence properties of wound-associated bacteria cultured under various temperature, salinity, and pH conditions
title_full Autofluorescence properties of wound-associated bacteria cultured under various temperature, salinity, and pH conditions
title_fullStr Autofluorescence properties of wound-associated bacteria cultured under various temperature, salinity, and pH conditions
title_full_unstemmed Autofluorescence properties of wound-associated bacteria cultured under various temperature, salinity, and pH conditions
title_short Autofluorescence properties of wound-associated bacteria cultured under various temperature, salinity, and pH conditions
title_sort autofluorescence properties of wound associated bacteria cultured under various temperature salinity and ph conditions
topic Bacterial autofluorescence
Endogenous porphyrins
Temperature
NaCl concentration
pH
url https://doi.org/10.1186/s12866-025-04200-3
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