Outbreaks of foodborne diseases are regularly reported worldwide. In particular, uncooked plant food is considered risky in terms of microbiological safety. Food is also the most important transmission route for resistant microorganisms from animals to humans. Photodynamic Decontamination (PDc) of foodstuff was recently introduced as a novel approach for increasing microbiological food safety. We investigated the efficiency of PDc on plant food with different geometries (flat, spherical and complex) using a two-dimensional LED array as a light source (435 nm, 33.8 J cm−2) and the cationic curcumin derivative SACUR-3 as a photosensitiser. A photoantibacterial effect (>3 log10 CFU reduction) was achieved on all flat substrates (slices of cucumber, tomato and lettuce) with 10 μM, 50 μM or 100 μM SACUR-3. The maximal photokilling with a relative inactivation of 5.6 log10 was measured on lettuce using 50 μM of the photoactive compound. Phototreatment of non-germinated fenugreek seeds and mung beans was successful if the spherical objects were rotated while under illumination (antibacterial effect at 100 μM SACUR-3). The decontamination of mung bean germlings with a more complex geometry using the PDc approach was ineffective with the two-dimensional light source. In conclusion, PDc based on the cationic curcumin derivative SACUR-3 is very effective at improving the microbiological safety of plant food with a flat or spherical geometry. More complex objects will require the development of novel illumination devices.
5 June 2014 in Photochemical & Photobiological Sciences
Increasing antibiotic resistance is one of the world's greatest health problems. The food chain is an important factor in the transfer of resistant germs from animals to humans. This study focuses on photodynamic inactivation (PDI), employing curcumin bound to polyvinylpyrrolidone (PVP-C) and NovaSol®-curcumin as photosensitizers, as potent tool for the decontamination of cucumber, pepper and chicken meat from Staphylococcus aureus (serving as the model for methicillin-resistant S. aureus, MRSA). Both curcumin and PVP have been approved as food additives, consequently exhibiting excellent biocompatibility. Vegetables and meat were contaminated with S. aureus and sprinkled with PVP-C and NovaSol®-curcumin at concentrations of 50 and 100 μM, respectively. Illumination was performed immediately using visible light (435 nm, 9.4 mW cm−2, 33.8 J cm−2). The PDI efficiency was determined by quantitative analyses of colony forming units 24 h post illumination. Additionally, the long-term effects of the photodynamic inactivation on cucumbers were investigated by quantitative analyses of the viable bacterial fraction after 24 and 48 h. Photodynamic inactivation of S. aureus revealed a mean reduction of 2.6 log10 (99.8%) for cucumbers, 2.5 log10 (99.7%) for pepper and 1.7 log10 (98%) for chicken meat relative to control samples. The bactericidal effect compared to controls seems to last for at least 48 h. Furthermore, no visible changes of the exterior appearance of foodstuff after photodynamic decontamination were observed. Photodynamic inactivation may therefore constitute a safe, economic and effective decontamination technique, which is harmless to health and not noticeable to consumers.
3 February 2010 in Photochemical & Photobiological Sciences
Antibacterial photodynamic treatment (APDT) might prove valuable as an alternative to the application of chemoantibiotics for the treatment of staphylococcal infections. The rapid uptake of the photosensitizing agent into bacteria allows for selective killing of microorganisms whilst sparing the eukaryotic host tissue, but also requires removal of excessive dye after the incubation period. We tested water-soluble formulations of hypericin (PVP-hypericin) and m-tetrahydroxyphenylchlorin (Fospeg®), which could be applied as aqueous sprays and removed easily by flushing with buffers, for their efficiency in killing Staphylococcus aureus. For both sensitizers, 100 nM of the photoactive substance incubated for 5 min and illuminated for 30 min at 75 mW cm−2 lead to a 4–5 log unit reduction in bacterial count. At a concentration of 300 nM (incubation time 5 min), 30 min illumination at 25 mW cm−2 is more effective than 10 min illumination at 75 mW cm−2 (both resulting in the same fluence). We suggest both substances as promising candidates for treatment of staphylococcal infections in wounds with APDT.
5 July 2013 in Photochemical & Photobiological Sciences
Photodynamic inactivation (PDI), the light-induced and photosensitizer-mediated overproduction of reactive oxygen species in microorganisms, represents a convincing approach to treat infections with (multi-resistant) pathogens. Due to its favourable photoactive properties combined with excellent biocompatibility, curcumin derived from the roots of turmeric (Curcuma longa) has been identified as an advantageous photosensitizer for PDI. To overcome the poor water solubility and the rapid decay of the natural substance at physiological pH, we examined the applicability of polyvinylpyrrolidone curcumin (PVP-C) in an acidified aqueous solution (solubility of PVP-C up to 2.7 mM) for photoinactivation of Gram(+) and Gram(−) bacteria. Five micromolar PVP-C incubated for 5 minutes and illuminated using a blue light LED array (435 ± 10 nm, 33.8 J cm−2) resulted in a >6 log10 reduction of the number of viable Staphylococcus aureus. At this concentration, longer incubation periods result in a lower phototoxicity, most likely due to degeneration of curcumin. Upon an increase of the PVP-C concentration to 50 μM (incubation for 15 or 25 min) a complete eradication of Staphylococcus aureus can be achieved. As expected for a non-cationic photosensitizer, cell wall permeabilization with CaCl2 prior to addition of 50 μM PVP-C for 15 min is necessary to induce a drop in the count of the Gram(−) Escherichia coli for more than 3 log10. As both constituents of the formulation, curcumin (E number E100) and polyvinylpyrrolidone (E1201), have been approved as food additives, a PDI based on PCP-C might allow for a very sparing clinical application (e.g. for disinfection of wounds) or even for employment in aseptic production of foodstuffs.
In vitroresearch performed on eukaryotic or prokaryotic cell cultures usually represents the initial step for characterization of a novel photosensitizer (PS) intended for application in photodynamic therapy (PDT) of cancer or photodynamic inactivation (PDI) of microorganisms. Although many experimental steps of PS testing make use of the wide spectrum of methods readily employed in cell biology, special aspects of working with photoactive substances, such as the autofluorescence of the PS molecule or the requirement of light protection, need to be considered when performingin vitroexperiments in PDT/PDI. This tutorial represents a comprehensive collection of operative instructions, by which, based on photochemical and photophysical properties of a PS, its uptake into cells, the intracellular localization and photodynamic action in both tumor cells and microorganisms novel photoactive molecules may be characterized for their suitability for PDT/PDI. Furthermore, it shall stimulate the efforts to expand the convincing benefits of photodynamic therapy and photodynamic inactivation within both established and new fields of applications and motivate scientists of all disciplines to get involved in photodynamic research.
19 June 2019 in Photochemical & Photobiological Sciences
The ever growing world-population poses challenges concerning the need for more food free of pesticide residues. The most common means to control plant pathogens is through the application of pesticides, which raises concerns over safety for humans and the environment. Recently, Photodynamic Inactivation (PDI) of microorganisms using natural photosensitizers has shown itself to be a powerful tool to combat bacteria and fungi. This study investigates the efficacy of PDI against the Gram(+) bacterial plant pathogen Rhodococcus fascians and Gram(−) Xanthomonas axonopodis and Erwinia amylovora using two chlorin e6 derivatives as photosensitizers: anionic sodium magnesium chlorophyllin (Chl, approved as food additive E140) in combination with cell wall permeabilizing agents (Na2EDTA or Polyaspartic acid sodium salt (PA)) and B17-0024, a mixture of chlorin e6 derivatives with cationic moieties at physiological pH. Both photosensitizers show excellent efficacy against R. fascians, whereby B17-0024 is phototoxic at a one order of magnitude lower concentration than Chl (10 μM B17-0024: relative inactivation (r.i.) >7.5 × 106, 100 μM Chl: r.i. 2.2 × 106, illumination with 26.6 J cm−2, 395 nm). The phototreatment of Gram(−) bacteria with Chl requires the obligatory use of cell wall permeabilizing agents like Na2EDTA (X. axonopodis) or PA (E. amylovora) to induce significant killing (more than 7 log units at 100 μM). On the other hand, B17-0024 proves to be a highly effective photosensitizer inducing bacterial inactivation at very low concentrations (10 μM for R. fascians and X. axonopodis, 100 μM for E. amylovora) without additives. In summary, PDI using both the natural photosensitizer Chl in combination with cell wall permeabilizing agents is effective and environmentally friendly. As an alternative, B17-0024 is highly photoactive against all model strains tested – even without cell wall permeabilizing agents. The photodynamic approach based on chlorin e6 derivatives should add to the growers’ toolbox as a preferred alternative for the control of phytopathogens.