Approximately 70% of the terrestrial area is covered with water, but only a small water fraction is compatible with terrestrial life forms. Due to the increment in human consumption, the need for water resources is increasing, and it is estimated that more than 40% of the population worldwide will face water stress/scarcity within the next few decades. Water recycling and reuse may offer the opportunity to expand water resources. For that, the wastewater treatment paradigm should be changed and adequately treated wastewater should be seen as a valuable resource instead of a waste product. It is easily understandable that the exact composition and constituent concentration of wastewater vary according to its different sources (industrial, agricultural, urban usage of water). Consequently, a variety of known and emerging pollutants like heavy metals, antibiotics, pesticides, phthalates, polyaromatic hydrocarbons, halogenated compounds and endocrine disruptors have been found in natural water reservoirs, due to the limited effectiveness of conventional wastewater treatment. The conventional approach consists of a combination of physical, chemical and biological processes, aiming at the removal of large sediments such as heavier solids, scum and grease and of organic content in order to avoid the growth of microorganisms and eutrophication of the receiving water bodies. However, this approach is not sufficient to reduce the chemical pollutants and much less the emerging chemical pollutants. In this review, after some considerations concerning chemical pollutants and the problematic efficiency of their removal by conventional methods, an update is presented on the successes and challenges of novel approaches for wastewater remediation based on advanced oxidation processes. An insight into wastewater remediation involving the photodynamic approach mediated by tetrapyrrolic derivatives will be underlined.

Staphylococcus aureus is a gram-positive bacterium who integrates the human microbiota. Nevertheless, these bacteria can be pathogenic to the humans. Due to the increasing occurrence of antibiotic-resistant S. aureus strains, new approaches to control this pathogen are necessary. The antimicrobial photodynamic inactivation (PDI) process is based in the combined use of light, oxygen and an intermediary agent (a photosensitizer). These three components interact to generate cytotoxic reactive oxygen species that irreversibly damage vital constituents of the microbial cells and ultimately lead to cell death. Although PDI is being shown to be a promising alternative to the antibiotic approach for the inactivation of pathogenic microorganisms, information on effects of photosensitization on particular virulence factors is strikingly scarce. The objective of this work was to evaluate the effect of PDI on virulence factors of S. aureus and to assess the potential development of resistance of this bacterium as well as the recovery of the expression of the virulence factors after successive PDI cycles. For this, the photosensitizer 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin tetra-iodide (Tetra-Py+-Me) and six strains of S. aureus (one reference strain, one strain with 1 enterotoxin, two strains with 3 enterotoxins and two strains methicillin resistant (MRSA) – one with 5 enterotoxins and the other without enterotoxins) were used. The effect of photosensitization on catalase activity, beta hemolysis, lipases, thermonuclease, enterotoxins, coagulase production and resistance/susceptibility to methicillin was tested. To assess the development of resistance after successive cycles of treatment, three strains of S. aureus (ATCC 6538, 2065 MA and SA 3 MRSA) were used. The surviving colonies of a first cycle of PDI were collected from the solid medium and subjected to further nine consecutive cycles of PDI. The results indicate that the expression of some external virulence factors is affected by PDI and enterotoxin producing strains are more susceptible to PDI than non-toxigenic strains. The surviving bacteria neither developed resistance nor recovered the expression of the virulence factors after 10 cycles of treatment. PDI, contrarily to traditional antibiotics, inhibits the expression of virulence factors, inactivating even more efficiently highly virulent strains than low virulent S. aureus strains, inactivating also antibiotic susceptible and

Wastewater (WW) from urban and industrial activities is often contaminated with microorganisms and chemical pollutants. To reduce the concentration of microorganisms in WW to levels comparable to those found in natural waters, the sewage effluent is usually subjected to disinfection with chlorine, ozone, or ultraviolet light, which may lead to the formation of toxic products and contribute to the selection of resistant genes. Moreover, the changing patterns of infectious diseases and the emerging of multidrug resistant microbial strains entail the development of new technologies for WW decontamination. Microbial photodynamic inactivation (PDI) with photosensitizers, oxygen, and visible light has demonstrated to be effective in the inactivation of microorganisms via photogeneration of reactive oxygen species able to induce microbial damage at the external structures level. The promising results of PDI suggest that this principle can be applied to WW treatment to inactivate microorganisms but also to photodegrade chemical pollutants. The aim of this study was to assess the applicability of PDI for the microbial and chemical decontamination of secondarily treated WW. To evaluate the efficiency of bacterial inactivation in WW, experiments were done in both phosphate buffer saline (PBS) and filtered WW with the bioluminescent Escherichia coli, using small and large volumes of WW. The potential of PDI to inactivate the native bacteria (E. coli and Enterococcus) present in WW was tested and assays without the adding of bacteria to the WW were performed. It was also tested if the same PDI protocol was able to induce phototransformation of phenol. The cationic porphyrin 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin tetra-iodide (Tetra-Py⁺-Me) was shown to be effective against both bacterial groups representing both Gram-negative and Gram-positive bacteria used as microbiological parameters to instigate water quality and even showing the power to photooxidate organic compounds. As the photosensitizer when immobilized on solid matrixes can be easily removed, recovered, and reused, an effective, less-expensive, easy-applicable, and environmentally friendly technology can be applied to treat WW, inactivating microorganisms and degrading chemical contaminants at the same time.

Pathogenic viruses are frequently present in marine and estuarine waters, due to poor wastewater (WW) treatments, which consequently affect water quality and human health. Chlorination, one of the most common methods used to ensure microbiological safety in tertiarily treated effluents, may lead to the formation of toxic chemical disinfection by-products on reaction with organic matter present in the effluents. Antimicrobial photodynamic therapy (aPDT) can be a promising disinfecting approach for the inactivation of pathogens, without the formation of known toxic by-products. Additionally, some studies have reported the potentiator effect on aPDT of some compounds, such as potassium iodide (KI) and hydrogen peroxide (H₂O₂). In the present study, the aPDT efficiency of a PS formulation constituted of five cationic porphyrins (Form) in the inactivation of E. coli T4-like bacteriophage, a model of mammalian viruses, in different aqueous matrices with different organic matter content, was evaluated. Photoinactivation studies were performed at different concentrations of Form and in the presence of the adjuvants KI and H₂O₂. The results showed that the efficiency of bacteriophage photoinactivation is correlated with the Form concentration, the amount of the organic matter in WW, and the adjuvant type. Form can be an effective alternative to controlling viruses in WW, particularly if combined with H₂O₂, allowing to significantly reduce PS concentration and treatment time. When combined with KI, the Form is less effective in inactivating T4-like bacteriophage in WW.

The last two years have been marked by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. This virus is found in the intestinal tract; it reaches wastewater systems and, consequently, the natural receiving water bodies. As such, inefficiently treated wastewater (WW) can be a means of contamination. The currently used methods for the disinfection of WW can lead to the formation of toxic compounds and can be expensive or inefficient. As such, new and alternative approaches must be considered, namely, photodynamic inactivation (PDI). In this work, the bacteriophage φ6 (or, simply, phage φ6), which has been used as a suitable model for enveloped RNA viruses, such as coronaviruses (CoVs), was used as a model of SARS-CoV-2. Firstly, to understand the virus’s survival in the environment, phage φ6 was subjected to different laboratory-controlled environmental conditions (temperature, pH, salinity, and solar and UV-B irradiation), and its persistence over time was assessed. Second, to assess the efficiency of PDI towards the virus, assays were performed in both phosphate-buffered saline (PBS), a commonly used aqueous matrix, and a secondarily treated WW (a real WW matrix). Third, as WW is generally discharged into the marine environment after treatment, the safety of PDI-treated WW was assessed through the determination of the viability of native marine water microorganisms after their contact with the PDI-treated effluent. Overall, the results showed that, when used as a surrogate for SARS-CoV-2, phage φ6 remains viable in different environmental conditions for a considerable period. Moreover, PDI proved to be an efficient approach in the inactivation of the viruses, and the PDI-treated effluent showed no toxicity to native aquatic microorganisms under realistic dilution conditions, thus endorsing PDI as an efficient and safe tertiary WW disinfection method. Although all studies were performed with phage φ6, which is considered a suitable model of SARS-CoV-2, further studies using SARS-CoV-2 are necessary; nevertheless, the findings show the potential of PDI for controlling SARS-CoV-2 in WW.

Antimicrobial photodynamic treatment (aPDT) offers an alternative option for combating microbial pathogens, and in this way, addressing the challenges of growing antimicrobial resistance. In this promising and effective approach, cationic porphyrins and related macrocycles have emerged as leading photosensitizers (PS) for aPDT. In general, their preparation occurs via N-alkylation of nitrogen-based moieties with alkyl halides, which limits the ability to fine-tune the features of porphyrin-based PS. Herein, is reported that the conjugation of porphyrin macrocycles with triphenylphosphonium units created a series of effective cationic porphyrin-based PS for aPDT. The presence of positive charges at both the porphyrin macrocycle and triphenylphosphonium moieties significantly enhances the photodynamic activity of porphyrin-based PS against both Gram-positive and Gram-negative bacterial strains. Moreover, bacterial photoinactivation is achieved with a notable reduction in irradiation time, exceeding 50%, compared to 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin (TMPyP), used as the reference and known as good PS. The improved capability of the porphyrin macrocycle to generate singlet oxygen combined with the enhanced membrane interaction promoted by the presence of triphenylphosphonium moieties represents a promising approach to developing porphyrin-based PS with enhanced photosensitizing activity.