Paraburkholderia fungorum Photoinactivation by Different Wavelengths

<i>Paraburkholderia fungorum</i> (<i>P. fungorum</i>) 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 <i>P. fungorum</i> 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, <i>p</i> < 0.001) with ROS generation as a more powerful predictor of inactivation than fluence dose alone (F = 60.6, <i>p</i> < 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 <i>P. fungorum</i> 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.