Antimicrobial Photodynamic Inactivation Research Articles

Efflux Pump Inhibitor Potentiates the Antimicrobial Photodynamic Inactivation of Multidrug-Resistant Acinetobacter baumannii.

Background: Acinetobacter baumannii, a nosocomial pathogen, poses a major public health problem due to generating resistance to several antimicrobial agents. Antimicrobial photodynamic inactivation (APDI) employs a nontoxic dye as a photosensitizer (PS) and light to produce reactive oxygen species that destroy bacterial cells. The intracellular concentration of PS could be affected by factors such as the function of efflux pumps to emit PS from the cytosol. Objective: To evaluate the augmentation effect of an efflux pump inhibitor, verapamil, three multidrug-resistant A. baumannii were subjected to APDI by erythrosine B (EB). Methods and results: The combination of EB and verapamil along with irradiation at 530 nm induced a lethal effect and more than 3 log colony-forming unit reduction to all A. baumannii strains in planktonic state. In contrast, EB and irradiation alone could produce only a sublethal effect on two of the strains. Conclusions: These data suggest that verapamil increases the intracellular concentration of EB, which potentiates the lethal efficacy of APDI. Verapamil could be applied with EB and green light to improve their antimicrobial efficacy against A. baumannii-localized infections.

On the Possibility of Using 5-Aminolevulinic Acid in the Light-Induced Destruction of Microorganisms.

Antimicrobial photodynamic inactivation (aPDI) is a method that specifically kills target cells by combining a photosensitizer and irradiation with light at the appropriate wavelength. The natural amino acid, 5-aminolevulinic acid (5-ALA), is the precursor of endogenous porphyrins in the heme biosynthesis pathway. This review summarizes the recent progress in understanding the biosynthetic pathways and regulatory mechanisms of 5-ALA synthesis in biological hosts. The effectiveness of 5-ALA-aPDI in destroying various groups of pathogens (viruses, fungi, yeasts, parasites) was presented, but greater attention was focused on the antibacterial activity of this technique. Finally, the clinical applications of 5-ALA in therapies using 5-ALA and visible light (treatment of ulcers and disinfection of dental canals) were described.

Design of polymeric carriers to enhance antimicrobial photodynamic inactivation

Mixed polymeric micelles are a promising tool for the design of nanocarriers with pharmaceutical properties. These micellar systems allow reversing numerous properties of lipophilic drugs. Particularly, when applied to transport photosensitizers (PSs), they give rise to a wide variety of nanophotosensitizers (nPSs). These compounds are used to inactivate numerous bacteria resistant to conventional antibiotics. This research proposes the development of new mixed micelles, with better properties than simple micelles, for the trapping of Neutral Red (NR) and its monobrominated derivative (NRBr). Based on the combination of Pluronic P-123 with Pluronic F-108, Pluronic L-61 and Tween 80, 15 binary micellar systems were obtained. These mixed micelles presented a single population of homogeneous particles, with a diameter less than 70 nm. Moreover, several mixed micelles show smaller CMC than the simple system. These properties are predictors of high micellar stability and good anti-dilution properties, favoring their drug delivery applications. For these reasons, were loaded with NR and NRBr, showing good encapsulation efficiency. The 30 new nPSs presented an adequate size for pharmaceutical use and markedly enhanced the photochemical reactivity of the free PSs. Many of the mixed micellar systems showed greater production of singlet oxygen than the simple micelles of Pluronic P-123. Furthermore, all micellar systems increased the chemical stability of NRBr, which degrades in pH 7.4 buffer solution. Finally, all nPSs presented greater phototoxic activity against methicillin-resistant Staphylococcus aureus than free PSs. The nPSs formed by P-123: F-108 (90:10) + NR, P-123: F-108 (90:10) + NRBr, P-123: L-61 (80:20) + NRBr and P −123: T80 (80:20) + NRBr caused greater photodynamic inactivation of the microorganism than simple P-123 micellar systems. These nPSs could be considered excellent candidates for application in the treatment of infections caused by bacteria resistant to antibiotics.

Antimicrobial Potency of Fmoc-Phe-Phe Dipeptide Hydrogels with Encapsulated Porphyrin Chromophores Is a Promising Alternative in Antimicrobial Resistance.

Antimicrobial resistance (AMR) poses a significant global health risk as a consequence of misuse of antibiotics. Owing to the increasing antimicrobial resistance, it became imperative to develop novel molecules and materials with antimicrobial properties. Porphyrins and metalloporphyrins are compounds which present antimicrobial properties especially after irradiation. As a consequence, porphyrinoids have recently been utilized as antimicrobial agents in antimicrobial photodynamic inactivation in bacteria and other microorganisms. Herein, we report the encapsulation of porphyrins into peptide hydrogels which serve as delivery vehicles. We selected the self-assembling Fmoc-Phe-Phe dipeptide, a potent gelator, as a scaffold due to its previously reported biocompatibility and three different water-soluble porphyrins as photosensitizers. We evaluated the structural, mechanical and in vitro degradation properties of these hydrogels, their interaction with NIH3T3 mouse skin fibroblasts, and we assessed their antimicrobial efficacy against Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Escherichia coli (E. coli) bacteria. We found out that the hydrogels are cytocompatible and display antimicrobial efficiency against both strains with the zinc porphyrins being more efficient. Therefore, these hydrogels present a promising alternative for combating bacterial infections in the face of growing AMR concerns.

Antimicrobial photodynamic inactivation pH-responsive films based on gelatin/chitosan incorporated with aloe-emodin

Using novel active food packaging has gradually become a daily necessity in terms of impeding microbial contamination. Here, an antimicrobial photodynamic inactivation (PDI) pH-responsive film is developed by incorporating aloe-emodin (AE) into a vehicle of gelatin/chitosan (GC). Besides enhancement in hydrophobicity, the well-dispersed crystals of AE in the GC matrix by hydrogen bonding can upgrade the film's mechanical strength and barrier. The matrix is capable of regulating the release of AE in response to acidic stimuli by a combination mechanism of diffusion and polymer relaxation. Being benefitted from the inherent bioactivity of AE and the PDI activity under visible light irradiation (i.e., 456 nm), the target film of GC-AE2 has excellent antibacterial effect towards Staphylococcus aureus and Escherichia coli, showing bacterial viability of 9.93 ± 1.33 % and 14.85 ± 1.16 %, respectively. Furthermore, the film can effectively thwart Botrytis cinerea infection in cherry tomatoes, demonstrating its potential in preventing the microbial spoilage of postharvest fruits.

APDI meets PPE: photochemical decontamination in healthcare using methylene blue-where are we now, where will we go?

Personal protective equipment (PPE) reuse, first recommended in the context of the SARS-CoV-2 pandemic, can mitigate shortages in crisis situations and can greatly reduce the environmental impact of typically single-use PPE. Prior to safe reuse, PPE must be sanitized and contaminating pathogens-in current circumstances viruses in particular-must be inactivated. However, many established decontamination procedures are not equitable and remain unavailable in low-resource settings. In mid-2020, an interdisciplinary consortium of researchers first studied the potential of implementing cheap and easy-to-use antimicrobial photodynamic inactivation (aPDI) using methylene blue as photosensitizer to decontaminate face masks and filtering facepiece respirators. In this perspective piece, we describe the development of this novel method, discuss recent advances, and offer insights into how equitable PPE decontamination via methylene blue-based aPDI may be integrated into circular economy policies in the healthcare sector.

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.

Phenylalanine-Arginine-β-Naphthylamide Enhances the Photobactericidal Effect of Methylene Blue on Pseudomonas aeruginosa.

Objective: To investigate the effectiveness, dosing sequence, concentration, and mechanism of antimicrobial photodynamic inactivation (aPDI) using methylene blue (MB) plus phenylalanine-arginine-β-naphthylamide (PAβN) against Pseudomonas aeruginosa. Methods: P. aeruginosa bacterial suspension was incubated with MB for different times (5-240 min), and then, 10 J/cm2 red light was irradiated. The efflux pump inhibitor (EPI) PAβN (10-100 μg/mL) was combined with MB (1-20 μM) in different sequences (PAβN-first, PAβN+MB, PAβN-after). Colony-forming units were then determined by serial dilution. Results: Using MB 10 μM plus 10 J/cm2, the killing effect of MB-aPDI on P. aeruginosa increased first and then decreased with longer incubation time. The killing effect of MB+PAβN-aPDI on P. aeruginosa was better than that of MB-aPDI (p < 0.05) by up to 2 logs. PAβN-first had the best killing effect, whereas PAβN-after had the worst killing effect. The killing effect increased with PAβN concentration and at 100 μg/mL reached 5.1 logs. Conclusions: The EPI PAβN enhanced the bactericidal effect of MB-aPDI on P. aeruginosa, especially when added before MB. It is proposed that MB is a substrate of the resistance-nodulation-division family efflux pump.

An alternative approach for daily perineal care of patients with indwelling urinary catheterization: Photodynamic inactivation with cationic porphyrin derivatives

Background: Catheter-associated urinary tract infections (CAUTI) constitute a significant portion of healthcare-associated infections. Using antiseptic for routine daily perineal care of patients with IUC may reduce CAUTIs. Aim: This study aimed to examine antimicrobial photodynamic inactivation (aPDI) against clinical isolates for use in the daily perineal care of patients with IUC. In addition, it was also aimed to compare the antimicrobial activities of aPDI and 0.1% chlorhexidine gluconate. Methods:  In this in-vitro study, cationic porphyrin derivatives (CPDs) were used as photosensitizers in the experiments. CPDs, named PM, PE, PN, and PL were synthesized by the researchers. A diode laser device emitting light with a wavelength of 450 nm (blue light) was used as the light source. Methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli and Klebsiella pneumoniae with multidrug-resistant (MDR) properties and Candida albicans were used.  Photosensitizer (PS), aPDI, light (L), and control (C) groups in aPDI experiments; control (C) and chlorhexidine gluconate 0.1% groups were used in the chlorhexidine gluconate experiments. Survival was calculated based on CFU/mL in the control group. Results: In experiments, combinations of 25 J/cm² with 6.25 and 3.125 µM PM, PE reduced E. coli, K. pneumoniae, MRSA, and C. albicans survival in the range of 8.70 to 11.53 log₁₀. In aPDI experiments performed with 6.25 and 3.125 µM PN and PL concentrations at the same energy density, reductions in the range of 4.41 to 0.17 log₁₀ were observed in all four clinical isolates. In experiments where 1.5625 µM concentration was used, survival decreased in the range of 8.29 to 10.87 log₁₀ in PM and PE, while antimicrobial activity was limited in PN and PL. In the 0.1% chlorhexidine gluconate experiments, the survival reduction in all four clinical isolates ranged from 8.87 to 10.24 log₁₀. Conclusion: For PM and PE, a very strong aPDI was obtained in C. albicans, E.coli, K. pneumoniae, and MRSA at low concentrations and energy density. The same antimicrobial activity was found in experiments using 0.1% chlorhexidine gluconate. In this context, we would like to inform you that aPDI to be performed with a combination of 25 J/cm² at 6.25 and 3.125 µM concentrations of PM and PE has the potential to be an antiseptic in the daily perineal care of patients with IUC.

Photoactivated Gallium Porphyrin Reduces Staphylococcus aureus Colonization on the Skin and Suppresses Its Ability to Produce Enterotoxin C and TSST-1.

Staphylococcus aureus is a key pathogen in atopic dermatitis (AD) pathogenicity. Over half of AD patients are carriers of S. aureus. Clinical isolates derived from AD patients produce various staphylococcal enterotoxins, such as staphylococcal enterotoxin C or toxic shock syndrome toxin. The production of these virulence factors is correlated with more severe AD. In this study, we propose cationic heme-mimetic gallium porphyrin (Ga3+CHP), a novel gallium metalloporphyrin, as an anti-staphylococcal agent that functions through dual mechanisms: a light-dependent mechanism (antimicrobial photodynamic inactivation, aPDI) and a light-independent mechanism (suppressing iron metabolism). Ga3+CHP has two additive quaternary ammonium groups that increase its water solubility. Furthermore, Ga3+CHP is an efficient generator of singlet oxygen and can be recognized by heme-target systems such as Isd, which improves the intracellular accumulation of this compound. Ga3+CHP activated with green light effectively reduced the survival of clinical S. aureus isolates derived from AD patients (>5 log10 CFU/mL) and affected their enterotoxin gene expression. Additionally, there was a decrease in the biological functionality of studied toxins regarding their superantigenicity. In aPDI conditions, there was no pronounced toxicity in HaCaT keratinocytes with both normal and suppressed filaggrin gene expression, which occurs in ∼50% of AD patients. Additionally, no mutagenic activity was observed. Green light-activated gallium metalloporphyrins may be a promising chemotherapeutic to reduce S. aureus colonization on the skin of AD patients.

Photodynamic Activity of Chlorophyllin and Polyethylenimine on Pseudomonas aeruginosa Planktonic, Biofilm and Persister Cells.

Antimicrobial photodynamic inactivation is considered a promising antimicrobial approach that may not develop resistance in the near future. Here, we investigate the influence of the photosensitizer chlorophyllin (CHL) and the cationic permeabilizer polyethylenimine (PEI), exposed to a red light-emitting diode, on the human pathogen Pseudomonas aeruginosa free-living planktonic cells, the sessile biofilm and persister cells. The broth microdilution checkerboard method was used to test antimicrobial susceptibility. As a substrate for biofilms, the Calgary biofilm device was used, and the quantification of the biofilm biomass was carried out using a crystal violet assay. Serine hydroxamate was used for the induction of persisters. Our findings reveal that PEI ameliorates the antimicrobial activity of CHL against P. aeruginosa planktonic and biofilm states, and the concentration required to eradicate the bacteria in the biofilm is more than fourfold that is required to eradicate planktonic cells. Interestingly, the persister cells are more susceptible to CHL/PEI (31.25/100 µg mL-1) than the growing cells by 1.7 ± 0.12 and 0.4 ± 0.1 log10 reduction, respectively, after 15 min of illumination. These data demonstrate that CHL excited with red light together with PEI is promising for the eradication of P. aeruginosa, and the susceptibility of P. aeruginosa to CHL/PEI is influenced by the concentrations and the exposure time.

Antimicrobial Photodynamic Inactivation: An Alternative for Group B Streptococcus Vaginal Colonization in a Murine Experimental Model.

Streptococcus agalactiae, referred to as Group B Streptococcus (GBS), is a prominent bacterium causing life-threatening neonatal infections. Although antibiotics are efficient against GBS, growing antibiotic resistance forces the search for alternative treatments and/or prevention approaches. Antimicrobial photodynamic inactivation (aPDI) appears to be a potent alternative non-antibiotic strategy against GBS. The effect of rose bengal aPDI on various GBS serotypes, Lactobacillus species, human eukaryotic cell lines and microbial vaginal flora composition was evaluated. RB-mediated aPDI was evidenced to exert high bactericidal efficacy towards S. agalactiae in vitro (>4 log10 units of viability reduction for planktonic and >2 log10 units for multispecies biofilm culture) and in vivo (ca. 2 log10 units of viability reduction in mice vaginal GBS colonization model) in microbiological and metagenomic analyses. At the same time, RB-mediated aPDI was evidenced to be not mutagenic and safe for human vaginal cells, as well as capable of maintaining the balance and viability of vaginal microbial flora. aPDI can efficiently kill GBS and serve as an alternative approach against GBS vaginal colonization and/or infections.

Photoantimicrobial activity of Schiff-base morpholino phthalocyanines against drug resistant micro-organisms in their planktonic and biofilm forms

Antimicrobial photodynamic inactivation (aPDI) is a treatment for the eradication of drug-resistant micro-organisms. One of the advantages of this technique, is that there is minimal possibility of microbial resistance. Hence, herein, the preparation and characterization of novel neutral and cationic morpholine containing Schiff base phthalocyanines are reported. The cationic complexes gave moderate singlet oxygen quantum yields (ΦΔ) of ∼0.2 in aqueous media. Conversely, the neutral complexes generated very low ΦΔ values making them very poor candidates for antimicrobial studies. The cationic phthalocyanines showed excellent photodynamic activity against planktonic cells of all micro-organisms (Candida albicans, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Salmonella enterica subspecies enterica serovar Choleraesuis, vancomycin-resistant Enterococcus faecium, and methicillin-resistant Staphylococcus aureus). The efficiency of aPDI was shown to be both concentration and light-dose-dependent. Mono biofilms were susceptible when treated with 200 µM of cationic Pcs at 108 J/cm2. However, ∼10% of the mixed biofilm survived after treatment.

Photosensitizer to the rescue: in planta and field application of photodynamic inactivation against plant pathogenic bacteria.

Control of plant pathogens using chemical and synthetic pesticides raises a major safety concern for humans and the environment. Despite the ongoing exploration of sustainable alternative methods, management practices for pathogens, especially bacteria, have remained almost unchanged over decades, whereby long-term uses of copper and antibiotics has led to widespread bacterial resistance in the field. Antimicrobial photodynamic inactivation (aPDI) of bacteria is emerging as an alternative strategy to combat resistant plant pathogens. aPDI utilizes light-sensitive molecules (photosensitizers) that upon illumination produce reactive oxygen species able to kill pathogens. Here we explore the potential of an anionic semisynthetic water-soluble derivative of chlorophyl (Sodium Magnesium Chlorophyllin: Mg-chl), as an antibacterial agent in planta, by simulating processes naturally occurring in the field. Mg-chl in combination with Na2EDTA (cell wall permeabilizing agent) was able to effectively inhibit Pseudomonas syringae pv. tomato DC3000 in vitro and in planta in both tomato and N. benthamiana. Notably, Mg-chl in combination with Na2EDTA and the common surfactant Morwet D-400 significantly reduced Xanthomonas hortorum pv. gardneri and Xanthomonas fragarie, respectively, in a commercial greenhouse trial against bacterial spot disease in tomato and in field experiments against angular leaf spot disease in strawberries.

Rose bengal-decorated rice husk-derived silica nanoparticles enhanced singlet oxygen generation for antimicrobial photodynamic inactivation.

The online version contains supplementary material available at 10.1007/s10853-023-08194-z.

Antimicrobial photodynamic inactivation of planktonic and biofilm cells by covalently immobilized porphyrin on polyethylene terephthalate surface

The appearance of resistant strains and the persistence of biofilms on different surfaces in a wide range of settings represent serious public health threats. Antimicrobial photodynamic inactivation (aPDI) is a promising alternative technology to overcome these challenges. The current study assessed the antimicrobial effect of polyethylene terephthalate (PET) discs covalently functionalized with a cationic porphyrin, against E. coli ATCC 25922 and Pseudomonas aeruginosa (Schroeter) Migula (ATCC® 10145™) growth. Irradiation with white LED light for 6 h resulted in 1.51 ± 0.03 and 3.26 ± 0.24 log reduction of planktonic P. aeruginosa and E. coli, respectively. The study also assessed the effect of the functionalized discs on biofilm formation by E. coli, P. aeruginosa, and Staphylococcus aureus subsp. aureus (ATCC® 6538P™). The biovolumes of S. aureus, P. aeruginosa, and E. coli biofilms were decreased by 0.6 ± 0.1, 0.56 ± 0.13 and 0.74 ± 0.06 log reduction, respectively. These results emphasize the ability of porphyrin-functionalized photoactive surfaces to kill bacterial cells and consequently prevent biofilm formation.

Antimicrobial photodynamic inactivation as an alternative approach to inhibit the growth of Cronobacter sakazakii by fine-tuning the activity of CpxRA two-component system.

Cronobacter sakazakii is an opportunistic foodborne pathogen primarily found in powdered infant formula (PIF). To date, it remains challenging to control the growth of this ubiquitous bacterium. Herein, antimicrobial photodynamic inactivation (aPDI) was first employed to inactivate C. sakazakii. Through 460 nm light irradiation coupled with hypocrellin B, the survival rate of C. sakazakii was diminished by 3~4 log. The photokilling effect was mediated by the attenuated membrane integrity, as evidenced by PI staining. Besides, scanning electron microscopy showed the deformed and aggregated cell cluster, and intracellular ROS was augmented by 2~3 folds when light doses increase. In addition to planktonic cells, the biofilm formation of C. sakazakii was also affected, showing an OD590nm decline from 0.85 to 0.25. In terms of molecular aspects, a two-component system called CpxRA, along with their target genes, was deregulated during illumination. Using the knock-out strain of ΔCpxA, the bacterial viability was reduced by 2 log under aPDI, a wider gap than the wildtype strain. Based on the promoted expression of CpxR and OmpC, aPDI is likely to play its part through attenuating the function of CpxRA-OmpC pathway. Finally, the aPDI system was applied to PIF, and C. sakazakii was inactivated under various desiccated or heated storage conditions. Collectively, aPDI serves as an alternative approach to decontaminate C. sakazakii, providing a new strategy to reduce the health risks caused by this prevalent foodborne pathogen.

Photodynamic toluidine blue-gold nanoconjugates as a novel therapeutic for Staphylococcal biofilms.

Staphylococci are among the most frequent bacteria known to cause biofilm-related infections. Pathogenic biofilms represent a global healthcare challenge due to their high tolerance to antimicrobials. In this study, water soluble polyethylene glycol (PEG)-coated gold nanospheres (28 ppm) and nanostars (15 ppm) with electrostatically adsorbed photosensitizer (PS) Toluidine Blue O (TBO) ∼4 μM were successfully synthesized and characterized as PEG-GNPs@TBO and PEG-GNSs@TBO. Both nanoconjugates and the TBO 4 μM solution showed remarkable, if similar, antimicrobial photodynamic inactivation (aPDI) effects at 638 nm, inhibiting the formation of biofilms by two Staphylococcal strains: a clinical methicillin-resistant Staphylococcus aureus (MRSA) isolate and Staphylococcus epidermidis (S. epidermidis) RP62A. Alternatively in biofilm eradication treatments, the aPDI effects of PEG-GNSs@TBO were more effective and yielded a 75% and 50% reduction in viable count of MRSA and S. epidermidis RP62A preformed biofilms, respectively and when compared with untreated samples. This reduction in viable count was even greater than that obtained through aPDI treatment using a 40 μM TBO solution. Confocal laser microscopy (CLSM) and scanning electron microscope (SEM) images of PEG-GNSs@TBO's aPDI treatments revealed significant changes in the integrity and morphology of biofilms, with fewer colony masses. The generation of reactive oxygen species (ROS) upon PEG-GNSs@TBO's aPDI treatment was detected by CLSM using a specific ROS fluorescent probe, demonstrating bright fluorescence red spots across the surfaces of the treated biofilms. Our findings shine a light on the potential synergism between gold nanoparticles (AuNPs) and photosensitizers in developing novel nanoplatforms to target Staphylococcal biofilm related infections.

Photodynamic inactivation of Staphylococcus aureus by ecological antibacterial solutions associating LED (ʎ 450 ± 10 nm) with curcumin and olive leaf extracts

Antimicrobial resistance is a problem in contemporary society, with Staphylococcus aureus standing out as a threat due to its ability to colonize, its pathogenicity, and its expression of several virulence factors. In this context, antimicrobial photodynamic inactivation (aPDI) emerges as an alternative to conventional microbicidal or microbiostatic systems, enabling numerous and successive applications without developing side effects and microbial resistance. In this context, an aPDI system against cultures of S. aureus based on a water-in-oil (W/O) emulsion incorporating curcumin as the photosensitizer (PS), with and without olive leaf extract (OLE), was developed and the antibacterial efficacy evaluated under LED activation (ʎ450±10nm) by depositing an energy density of 14J/cm2. The produced emulsified systems showed no significant differences in the droplet size and morphology, remaining stable along the tested period of 30days. The bacterial reduction achieved after the first aPDI application for the emulsions added with curcumin and curcumin combined with the OLE was 5 log10 CFU.mL-1 and 6 log10 CFU.mL-1, respectively, revealing a significant difference between the two groups (p<0.0001). After the second aPDI application, an increased microbial reduction (7 log10 CFU.mL-1) was observed for both studied groups even with a low significant difference (p<0.05). The PS loading through an emulsified system for aPDI obtained a bactericidal action against S. aureus, increased by applying two aPDI, showing a significant synergy between photodynamic inactivation, OLE delivery and antibacterial activity. In addition, the developed solutions were produced using natural products by an ecologically correct process.

Antimicrobial curcumin-mediated photodynamic inactivation of bacteria in natural bovine casing

Background: Outbreaks related to food contamination by resistant microorganisms is a worldwide concern that, motivates industries and research institutions to search for affordable solutions. Among the solutions that have been proposed, Photodynamic Inactivation (PDI) of microorganisms has gained prominence, among other aspects, because it is easy to apply and does not generate microbial resistance.Methods: In this study, we used the association between curcumin solubilized with Tween and light in the photodynamic inactivation process, using light-emitting diodes with a wavelength of 430 nm for decontamination S. Typhimurium and K. pneumoniae from bovine casings used as wrappers for meat products. The result was verified by counting and comparing the number of colony-forming units of the treatment concerning the negative control.Results: The solubilizer, Tween 80, used does not change the optical absorption of curcumin. An optical fluence of 150J/cm2 induces a microbial log reduction of 3.8±0.2 and 2.7±0.1 for S. Typhimurium, and K. pneumoniae contaminated guts, respectively. For the 200μM concentration of curcumin, the PDI provided a microbial log reduction of 3.16±0.03 for S. Typhimurium. For K. pneumoniae, the minimal inhibitory concentration of curcumin occurs up to 12.5μM, causing an microbial log reduction of 2.08±0.03.Conclusion: Both curcumin and tween are already used as additives in food production and do not pose health risks at the concentrations used. Furthermore, in the case of the material studied, the addition of curcumin favors the organoleptic quality associated with the color of the food, unlike the green or blue photossensitizers. The results pave the way for possible application of curcumin in finished meat products.