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Published on: April 6, 2022
Paraburkholderia fungorum Photoinactivation by Different Wavelengths
Robin Haag1,2, Martin Heßling1
1Institute of Medical Engineering and Mechatronics, Ulm University of Applied Sciences, Albert-Einstein-Allee 55, 89081 Ulm, Germany.
Abstract:
Paraburkholderia fungorum (P. fungorum) is an environmental bacterium with biotechnological applications, yet clinical isolations raise concerns about opportunistic infection risk. Genetically related pathogens exhibit substantial antibiotic resistance, motivating the investigation of alternative control strategies. This paper investigates P. fungorum photoinactivation across ultraviolet (222 nm, 254 nm, 313 nm, and 365 nm) and visible (400 nm and 464 nm) wavelengths including ROS (reactive oxygen species) quantification via DCFH-DA fluorescence assay. A two-way ANOVA analysis demonstrated that the wavelength is the dominant determinant of photoinactivation efficacy (F = 100.4, p < 0.001) with ROS generation as a more powerful predictor of inactivation than fluence dose alone (F = 60.6, p < 0.001) at 365 nm, 400 nm, and 464 nm. Ultraviolet irradiation at 254 nm achieved the highest efficiency (5.4 log reduction at 24 mJ/cm2), while 365 nm irradiation demonstrated a high efficacy of 5.2 log reduction at 122 J/cm2 with extraordinary ROS production (12,642-fold fluorescence increase). Conversely, inactivation efficiency declined at 400 nm (4.8 log reduction at 378 J/cm2 with 122-fold ROS increase) and 464 nm (3.4 log reduction at 3017 J/cm2 with lesser ROS detection at 27-fold increase). Wavelength-dependent ROS production correlated directly with bacterial inactivation efficacy, explaining the approximately 500-fold ROS differential between 365 nm and 464 nm. The demonstrated photosensitivity of P. fungorum across multiple wavelengths, with the statistical validation of wavelength-dependent mechanisms, provides a foundation for developing practical, mechanism-based phototherapy protocols tailored to specific clinical and environmental decontamination scenarios.
Insights
Investigating Paraburkholderia fungorum photoinactivation revealed that light wavelength significantly impacts bacterial reduction. Reactive oxygen species (ROS) generation, driven by specific wavelengths, is a key factor in this inactivation process.
Area of Science:
- Microbiology
- Photobiology
- Biotechnology
Background:
- Paraburkholderia fungorum is an environmental bacterium with biotechnological uses.
- Clinical isolation of P. fungorum raises concerns due to potential opportunistic infections.
- Antibiotic resistance in related pathogens necessitates alternative control strategies.
Purpose of the Study:
- To investigate the photoinactivation of Paraburkholderia fungorum across various UV and visible light wavelengths.
- To quantify reactive oxygen species (ROS) generation during photoinactivation.
- To determine the relationship between wavelength, ROS production, and bacterial inactivation efficacy.
Main Methods:
- Photoinactivation assays using ultraviolet (222, 254, 313, 365 nm) and visible (400, 464 nm) light.
- Quantification of reactive oxygen species (ROS) using the DCFH-DA fluorescence assay.
- Statistical analysis using two-way ANOVA to determine the significance of wavelength and fluence dose.
Main Results:
- Wavelength was the dominant factor in photoinactivation efficacy (p < 0.001).
- 254 nm UV achieved the highest inactivation (5.4 log reduction at 24 mJ/cm²).
- 365 nm irradiation showed high efficacy (5.2 log reduction at 122 J/cm²) with significant ROS production (12,642-fold increase).
- ROS generation strongly correlated with bacterial inactivation across tested wavelengths.
Conclusions:
- Paraburkholderia fungorum exhibits significant photosensitivity across multiple wavelengths.
- Wavelength-dependent ROS production is a primary mechanism for P. fungorum photoinactivation.
- Findings support the development of targeted phototherapy protocols for clinical and environmental decontamination.

