Photodynamic inactivation of methicillin-resistant Staphylococcus aureus by using Giemsa dye as a photosensitizer.

The rise of antibiotic-resistant bacteria calls for innovative approaches to combat multidrug-resistant strains. Here, the potential of the standard histological stain, Giemsa, to act as a photosensitizer (PS) for antimicrobial photodynamic inactivation (aPDI) against methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) strains is reported. Bioassays were performed using various Giemsa concentrations (ranging from 0.0 to 20.0 µM) under 625 nm illumination at a light dose of 30 J cm-2. Remarkably, Giemsa completely inhibited the growth of MSSA and MRSA bacterial colonies for concentrations at 10 µM and higher but exhibited no inhibitory effect without light exposure. Partition coefficient analysis revealed Giemsa's affinity for membranes. Furthermore, we quantified the production of reactive oxygen species (ROS) and singlet oxygen (1O2) to elucidate the aPDI mechanisms underlying bacterial inactivation mediated by Giemsa. These findings highlight Giemsa stain's potential as a PS in aPDI for targeting multidrug-resistant bacteria.

Photodynamic Inactivation of Methicillin-Resistant Staphylococcus aureus by a Natural Food Colorant (E-141ii).

This study evaluates the photosensitizing effectiveness of sodium copper chlorophyllin, a natural green colorant commonly used as a food additive (E-141ii), to inactivate methicillin-sensitive and methicillin-resistant Staphylococcus aureus under red-light illumination. Antimicrobial photodynamic inactivation (aPDI) was tested on a methicillin-sensitive reference strain (ATCC 25923) and a methicillin-resistant Staphylococcus aureus strain (GenBank accession number Mh087437) isolated from a clinical sample. The photoinactivation efficacy was investigated by exposing the bacterial strains to different E-141ii concentrations (0.0, 1.0, 2.5, 5.0, 10.0, and 20.0 µM) and to red light (625 nm) at 30 J cm-2. The results showed that E-141ii itself did not prevent bacterial growth for all tested concentrations when cultures were placed in the dark. By contrast, E-141ii photoinactivated both methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) under red-light illumination. However, different dose responses were observed for MSSA and MRSA. Whilst the MSSA growth was inhibited to the detection limit of the method with E-141ii at 2.5 µM, >10 µM concentrations were required to inhibit the growth of MRSA. The data also suggest that E-141ii can produce reactive oxygen species (ROS) via Type I reaction by electron transfer from its first excited singlet state to oxygen molecules. Our findings demonstrate that the tested food colorant has great potential to be used in aPDI of MRSA.

Effective killing of bacteria under blue-light irradiation promoted by green synthesized silver nanoparticles loaded on reduced graphene oxide sheets.

Graphene oxide (GO) materials loaded with silver nanoparticles (AgNPs) have drawn considerable attention due to their capacity to efficiently inactivate bacteria though a multifaceted mechanism of action, as well as for presenting a synergetic effect against bacteria when compared to the activity of AgNPs and GO alone. In this investigation, we present an inexpensive and environmentally-friendly method for synthesizing reduced GO sheets coated with silver nanoparticles (AgNPs/r-GO) using a coffee extract solution as a green reducing agent. The physical and chemical properties of the produced materials were extensively characterized by scanning electron microscopy (SEM), field-emission gun transmission electron microscopy (FEG-TEM), ultraviolet and visible absorption (UV-Vis), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), inductively coupled plasma-optical emission spectroscopy (ICP-OES) and ion release determination. The results demonstrated that AgNPs/r-GO composites were successfully produced, revealing the formation of micrometer-sized r-GO sheets decorated by AgNPs of approximately 70 nm diameter. Finally, bactericidal and photobactericidal effects of the AgNPs/r-GO composites were tested against Staphylococcus aureus, in which the results showed that the composites presented antimicrobial and photoantimicrobial activities. Moreover, our results demonstrated for the first time, to our knowledge, that an efficient process of bacterial inactivation can be achieved by using AgNPs/r-GO composites under blue light irradiation as a result of three different bacterial killing processes: (i) chemical effect promoted by Ag+ ion release from AgNPs; (ii) photocatalytic activity induced by AgNPs/r-GO composites, enhancing the bacterial photoinactivation due to the excited-Plasmons of the AgNPs when anchored on r-GO; and (iii) photodynamic effect produced by bacterial endogenous photosensitizers under blue-light irradiation. In summary, the present findings demonstrated that AgNPs/r-GO can be obtained by a non-toxic procedure with great potential for biomedical-related applications.

Evaluation of Eosin-Methylene Blue as a Photosensitizer for Larval Control of Aedes aegypti by a Photodynamic Process.

Aedes aegypti (Ae. aegypti) is a competent vector for transmitting important viral diseases such as yellow fever, dengue, chikungunya, and Zika. Several strategies have been applied to avoid Ae. aegypti proliferation by using environmental management, biological, and chemical approaches. However, the development of new methods for effective control of the insect vector population is still needed. Photodynamic control is an alternative way to control the vector population by using a physical approach based on the larval phototoxicity of a photosensitizer. In this context, the present study evaluated the use of eosin-methylene blue (EMB) as a new photosensitizer for photodynamic control of Ae. aegypti larval populations. The photodynamic assays were performed submitting Ae. aegypti third-instar larvae to different EMB concentrations (0.0, 0.5, 1.0, 5.0, 10.0, 50.0, and 100.0 µg mL-1) in combination of three different light doses (24.3, 48.6, and 97.2 J cm-2) under either white-light radiation from RGB LEDs or sunlight. The results demonstrated that EMB presented a rapid internalization into the larvae and was phototoxic. The photodynamic action induced 100% of larval mortality after about 40 min of sunlight irradiation even using low EMB concentration (0.5 µg mL-1). The findings reveal EMB as an effective photoactive compound to control larval populations of Ae. aegypti by photodynamic process induced by either sunlight or white-light from RGB LEDs.

Photoinactivation effect of eosin methylene blue and chlorophyllin sodium-copper against Staphylococcus aureus and Escherichia coli.

The use of eosin methylene blue according to Giemsa as photosensitizer is presented for the first time in this paper. The present study evaluated the potential application of chlorophyllin sodium copper salt (CuChlNa) and eosin methylene blue according to Giemsa (EMB) as antimicrobial photosensitizers (aPS) for photodynamic inactivation (PDI) of Staphylococcus aureus (gram-positive) and Escherichia coli (gram-negative) bacteria. The experiments were performed using S. aureus stain ATCC 25923 and E. coli ATCC 25922 in which five aPS concentrations (0.0, 1.0, 2.5, 5.0, 10.0, and 20.0 µM for S. aureus and 0.0, 5.0, 10.0, 20.0, 40.0, and 50.0 µM for E. coli) were prepared and added in 2 mL of a saline solution containing the bacterial inoculum. After aPS incubation, the samples were divided into two groups, one kept in the dark and another submitted to the illumination. Then, the bacterial inactivation was determined 18 h after the incubation at 37 °C by counting the colony-forming units (CFU). The results revealed that both EMB and CuChlNa can be used as aPS for the photoinactivation of S. aureus, while only EMB was able to photoinactivate E. coli. Nevertheless, a more complex experimental setup was needed for photoinactivation of E. coli. The data showed that EMB and CuChlNa presented similar photoinactivation effects on S. aureus, in which bacterial growth was completely inhibited at photosensitizer (PS) concentrations over 5 µM, when samples were previously incubated for 30 min and irradiated by a light dose of 30 J cm-2 as a result of an illumination of 1 h at 8.3 mW cm-2 by using a red light at 625 nm with a 1 cm beam diameter and output power of 6.5 mW. In the case of E. coli, bacterial growth was completely inhibited only when combining a PS incubation period of 120 min with concentrations over 20 µM.