Photodynamic inactivation of Escherichia coli by methylene blue and malachite green under red LED light

Abstract

This study assessed the effectiveness of methylene blue (MB) and malachite green (MG) on photodynamic inactivation (PDI) of Escherichia coli. The photosensitizers methylene blue (1000 mol L-1) and malachite green (250 mol L-1) were activated with a red light-emitting diode (LED) lamp (max = 636 nm). Bacterial suspensions containing 106 CFU mL-1 were irradiated for 5, 10 and 15 minutes (energy density = 119.9 J cm-2, 223.9 J cm-2 and 335.8 J cm-2, respectively). The following experimental conditions were performed for each photosensitizer: no light irradiation or photosensitizer, irradiation only, photosensitizer only or irradiation in the presence of a photosensitizer. Next, serial dilutions were prepared and seeded onto PCA medium for the determination of the number of colony-forming units per milliliter (CFU mL-1). The results were subjected to analysis of variance (ANOVA) and Tukey test (P<0.05). Photodynamic inactivation using MB and MG was effective in reducing the number of E. coli. Malachite green (250 µmol L-1) photosensitization was able to achieve reductions of over 89% in the viable counts after 15 min of irradiation and methylene blue (1000 µmol L-1), at the same conditions of irradiation, showed a rate growth inhibition of 94.6%. The red LED light used has proven to be effective in the photosensitizing dyes and proved a good alternative to conventional light sources such as laser.