Purification of living environments using photocatalysts: Inactivation of microorganisms and decomposition of allergens.

Many emerging and re-emerging infectious diseases are prevalent, and the number of patients with allergies is increasing. Therefore, the importance of purifying the living environment is increasing. Photocatalysts undergo extreme redox reactions and decompose organic matter upon exposure to the excitation light. In contrast to ultraviolet light and disinfectants, which are standard methods for inactivating viruses and eliminating microorganisms, photocatalysts can decompose toxic substances, such as endotoxins and allergens, rendering them harmless to the human body. Photocatalysts have attracted significant attention as potential antiviral and antimicrobial agents. This review outlines the antiviral, antimicrobial, and anti-allergenic effects of photocatalysts. Especially, we have discussed the inactivation of SARS-CoV-2 in liquids and aerosols, elimination of Legionella pneumophila in liquids, decomposition of its endotoxin, decomposition of cat and dog allergens, and elimination of their allergenicity using photocatalysts. Furthermore, we discuss future perspectives on how photocatalysts can purify living environments, and how photocatalytic technology can be applied to companion animals and the livestock industry.

Photocatalytic biocidal effect of copper doped TiO2 nanotube coated surfaces under laminar flow, illuminated with UVA light on Legionella pneumophila.

Legionella pneumophila can cause a potentially fatal form of humane pneumonia (Legionnaires' disease), which is most problematic in immunocompromised and in elderly people. Legionella species is present at low concentrations in soil, natural and artificial aquatic systems and is therefore constantly entering man-made water systems. The environment temperature for it's ideal growth range is between 32 and 42°C, thus hot water pipes represent ideal environment for spread of Legionella. The bacteria are dormant below 20°C and do not survive above 60°C. The primary method used to control the risk from Legionella is therefore water temperature control. There are several other effective treatments to prevent growth of Legionella in water systems, however current disinfection methods can be applied only intermittently thus allowing Legionella to grow in between treatments. Here we present an alternative disinfection method based on antibacterial coatings with Cu-TiO2 nanotubes deposited on preformed surfaces. In the experiment the microbiocidal efficiency of submicron coatings on polystyrene to the bacterium of the genus Legionella pneumophila with a potential use in a water supply system was tested. The treatment thus constantly prevents growth of Legionella pneumophila in presence of water at room temperature. Here we show that 24-hour illumination with low power UVA light source (15 W/m2 UVA illumination) of copper doped TiO2 nanotube coated surfaces is effective in preventing growth of Legionella pneumophila. Microbiocidal effects of Cu-TiO2 nanotube coatings were dependent on the flow of the medium and the intensity of UV-A light. It was determined that tested submicron coatings have microbiocidal effects specially in a non-flow or low-flow conditions, as in higher flow rates, probably to a greater possibility of Legionella pneumophila sedimentation on the coated polystyrene surfaces, meanwhile no significant differences among bacteria reduction was noted regarding to non or low flow of medium.

Peptidyl-Prolyl-cis/trans-Isomerases Mip and PpiB of Legionella pneumophila Contribute to Surface Translocation, Growth at Suboptimal Temperature, and Infection.

The gammaproteobacterium Legionella pneumophila is the causative agent of Legionnaires' disease, an atypical pneumonia that manifests itself with severe lung damage. L. pneumophila, a common inhabitant of freshwater environments, replicates in free-living amoebae and persists in biofilms in natural and man-made water systems. Its environmental versatility is reflected in its ability to survive and grow within a broad temperature range as well as its capability to colonize and infect a wide range of hosts, including protozoa and humans. Peptidyl-prolyl-cis/trans-isomerases (PPIases) are multifunctional proteins that are mainly involved in protein folding and secretion in bacteria. In L. pneumophila the surface-associated PPIase Mip was shown to facilitate the establishment of the intracellular infection cycle in its early stages. The cytoplasmic PpiB was shown to promote cold tolerance. Here, we set out to analyze the interrelationship of these two relevant PPIases in the context of environmental fitness and infection. We demonstrate that the PPIases Mip and PpiB are important for surfactant-dependent sliding motility and adaptation to suboptimal temperatures, features that contribute to the environmental fitness of L. pneumophila Furthermore, they contribute to infection of the natural host Acanthamoeba castellanii as well as human macrophages and human explanted lung tissue. These effects were additive in the case of sliding motility or synergistic in the case of temperature tolerance and infection, as assessed by the behavior of the double mutant. Accordingly, we propose that Mip and PpiB are virulence modulators of L. pneumophila with compensatory action and pleiotropic effects.

Inactivation kinetics and efficiencies of UV-LEDs against Pseudomonas aeruginosa, Legionella pneumophila, and surrogate microorganisms.

To demonstrate the effectiveness of UV light-emitting diodes (UV-LEDs) to disinfect water, UV-LEDs at peak emission wavelengths of 265, 280, and 300 nm were adopted to inactivate pathogenic species, including Pseudomonas aeruginosa and Legionella pneumophila, and surrogate species, including Escherichia coli, Bacillus subtilis spores, and bacteriophage Qß in water, compared to conventional low-pressure UV lamp emitting at 254 nm. The inactivation profiles of each species showed either a linear or sigmoidal survival curve, which both fit well with the Geeraerd's model. Based on the inactivation rate constant, the 265-nm UV-LED showed most effective fluence, except for with E. coli which showed similar inactivation rates at 265 and 254 nm. Electrical energy consumption required for 3-log10 inactivation (EE,3) was lowest for the 280-nm UV-LED for all microbial species tested. Taken together, the findings of this study determined the inactivation profiles and kinetics of both pathogenic bacteria and surrogate species under UV-LED exposure at different wavelengths. We also demonstrated that not only inactivation rate constants, but also energy efficiency should be considered when selecting an emission wavelength for UV-LEDs.

Control of Legionella Contamination and Risk of Corrosion in Hospital Water Networks following Various Disinfection Procedures.

Physical and chemical disinfection methods have been proposed with the aim of controlling Legionella water contamination. To date, the most effective procedures for reducing bacterial contamination have not yet been defined. The aim of this study was to assess the long-term effectiveness of various disinfection procedures in order to reduce both culturable and nonculturable (NC) legionellae in different hospital water networks treated with heat, chlorine dioxide, monochloramine, and hydrogen peroxide. The temperature levels and biocide concentrations that proved to give reliable results were analyzed. In order to study the possible effects on the water pipes, we verified the extent of corrosion on experimental coupons after applying each method for 6 months. The percentage of positive points was at its lowest after treatment with monochloramine, followed by chlorine dioxide, hydrogen peroxide, and hyperthermia. Different selections of Legionella spp. were observed, as networks treated with chlorine-based disinfectants were contaminated mainly by Legionella pneumophila serogroup 1, hyperthermia was associated with serogroups 2 to 14, and hydrogen peroxide treatment was associated mainly with non-pneumophila species. NC cells were detected only in heat-treated waters, and also when the temperature was approximately 60°C. The corrosion rates of the coupons were within a satisfactory limit for water networks, but the morphologies differed. We confirm here that chemical disinfection controls Legionella colonization more effectively than hyperthermia does. Monochloramine was the most effective treatment, while hydrogen peroxide may be a promising alternative to chlorine-based disinfectants due to its ability to select for other, less virulent or nonpathogenic species.

Effects of ultraviolet germicidal irradiation and swirling motion on airborne Staphylococcus aureus, Pseudomonas aeruginosa and Legionella pneumophila under various relative humidities.

UNLABELLED: Staphylococcus aureus, Pseudomonas aeruginosa, and Legionella pneumophila have been detected in indoor air and linked to human infection. It is essential to adopt control methods to inactivate airborne pathogens. By passing bioaerosols horizontally into a UV device at two flow rates (Qs) and moving cells around a central UVC lamp at relative humidity (RH) of 12.7-16.7%, 58.7-59.6%, and 87.3-90%, the effects of swirling motion and 254-nm ultraviolet germicidal irradiation (UVGI) against bioaerosols were assessed under UV-off and UV-on settings, respectively. An inverse relationship between RH and UVGI effectiveness was observed for every test bioaerosol (r = -0.74 ∼ -0.81, P < 0.0001). Increased UV resistance with RH is likely associated with the hygroscopicity of bioaerosols, evident by increased aerodynamic diameters at high RH (P < 0.05). UVGI effectiveness was significantly increased with decreasing Q (P < 0.0001). Moreover, P. aeruginosa was the most susceptible to UVGI, while the greatest UV resistance occurred in L. pneumophila at low RH and S. aureus at medium and high RH (P < 0.05). Results of UV off show P. aeruginosa and L. pneumophila were more sensitive to air-swirling motion than S. aureus (P < 0.05). Overall, test bioaerosols were reduced by 1.7-4.9 and 0.2-1.7 log units because of the UVGI and swirling movement, respectively. PRACTICAL IMPLICATIONS: The studied UV device, with a combination of swirling motion and UVGI, is effective to inactivate airborne S. aureus, P. aeruginosa, and L. pneumophila. This study also explores the factors governing the UVGI and swirling motion against infectious bioaerosols. With understanding the environmental and operational parameters, the studied UV device has the potential to be installed indoors where people are simultaneously present, for example, hospital wards and nursing homes, to prevent the humans from acquiring infectious diseases.

Induction of competence for natural transformation in Legionella pneumophila and exploitation for mutant construction.

Many Gram-positive and Gram-negative bacteria possess natural competence mechanisms for DNA -capture and internalization that play an important role in diversifying adaptation of bacteria through horizontal gene transfer. Natural transformation and other mechanisms of horizontal gene transfer are dependent on DNA recombination. Natural competence can be exploited both for studying adaptation and horizontal gene transfer as well as for genetic engineering of a strain. We report here different approaches to measure competence on solid and in liquid media by using a reporter plasmid where GFP is fused to the comEA gene or by inducing competence and measuring transformability induced by DNA-damaging stress. Finally we describe a method where competence is induced through a combined temperature and aeration shift, which may be exploited for the construction of mutants in Legionella pneumophila. This approach seems to be less prone to the appearance of secondary mutations during mutant construction as compared to procedures using electroporation.

Prevention of Legionnaires' disease in hospitals.

BACKGROUND: The first instance of Legionella infection in a Norwegian hospital was confirmed in 2005. We describe the best-known methods of eradicating Legionella in hospitals. MATERIALS AND METHOD: The article is based on the authors' experience of measures to prevent Legionnaires' disease in hospitals and on a non-systematic search in PubMed. RESULTS: There are several methods of combating Legionella in hospitals. These include chlorination, heat treatment, and the use of filters. However, recontamination easily re-occurs after eradication. The silver and copper ionisation treatment of water is a well-documented method for the systematic and long-term eradication of Legionella in water. The disadvantages of this method are that it is expensive, that there is a risk of discolouring the water, and that there is a possibility of developing resistance in environmental bacteria. This resistance mechanism can theoretically be transferred to bacteria that cause illness. INTERPRETATION: We recommend the silver and copper ionisation treatment of water as a method of preventing nosocomial Legionnaires' disease when standard methods fail and there is a high prevalence of Legionella in the water. The discolouration of operation instruments that occurs as a result of high silver concentrations can be avoided by using a separate water supply for operation units.

Antibiotics and UV radiation induce competence for natural transformation in Legionella pneumophila.

Natural transformation by competence is a major mechanism of horizontal gene transfer in bacteria. Competence is defined as the genetically programmed physiological state that enables bacteria to actively take up DNA from the environment. The conditions that signal competence development are multiple and elusive, complicating the understanding of its evolutionary significance. We used expression of the competence gene comEA as a reporter of competence development and screened several hundred molecules for their ability to induce competence in the freshwater living pathogen Legionella pneumophila. We found that comEA expression is induced by chronic exposure to genotoxic molecules such as mitomycin C and antibiotics of the fluoroquinolone family. These results indicated that, in L. pneumophila, competence may be a response to genotoxic stress. Sunlight-emitted UV light represents a major source of genotoxic stress in the environment and we found that exposure to UV radiation effectively induces competence development. For the first time, we show that genetic exchanges by natural transformation occur within an UV-stressed population. Genotoxic stress induces the RecA-dependent SOS response in many bacteria. However, genetic and phenotypic evidence suggest that L. pneumophila lacks a prototypic SOS response and competence development in response to genotoxic stress is RecA independent. Our results strengthen the hypothesis that competence may have evolved as a DNA damage response in SOS-deficient bacteria. This parasexual response to DNA damage may have enabled L. pneumophila to acquire and propagate foreign genes, contributing to the emergence of this human pathogen.

Photocatalytic treatment of bioaerosols: impact of the reactor design.

Comparing the UV-A photocatalytic treatment of bioaerosols contaminated with different airborne microorganisms such as L. pneumophila bacteria, T2 bacteriophage viruses and B. atrophaeus bacterial spores, pointed out a decontamination sensitivity following the bacteria > virus > bacterial spore ranking order, differing from that obtained for liquid-phase or surface UV-A photocatalytic disinfection. First-principles CFD investigation applied to a model annular photoreactor evidenced that larger the microorganism size, higher the hit probability with the photocatalytic surfaces. Applied to a commercial photocatalytic purifier case-study, the CFD calculations showed that the performances of the studied purifier could strongly benefit from rational reactor design engineering. The results obtained highlighted the required necessity to specifically investigate the removal of airborne microorganisms in terms of reactor design, and not to simply transpose the results obtained from studies performed toward chemical pollutants, especially for a successful commercial implementation of air decontamination photoreactors. This illustrated the importance of the aerodynamics in air decontamination, directly resulting from the microorganism morphology.

Ultraviolet light disinfection of hospital water for preventing nosocomial Legionella infection: a 13-year follow-up.

BACKGROUND AND OBJECTIVE: CDC has estimated that 23% of Legionella infections are nosocomial. When a new hospital was being constructed and a substantial increase in transplantation was anticipated, an ultraviolet light apparatus was installed in the water main of the new building because 27% of water samples from taps in the old hospital contained Legionella. This study reports the rate of nosocomial Legionella infection and water contamination since opening the new hospital. METHODS: Charts of all patients with positive Legionella cultures, direct immunofluorescent antibody (DFA), or urine antigen between April 1989 and November 2001 were reviewed. Frequencies of DFAs and urine antigens were obtained from the laboratory. RESULTS: None of the 930 cultures of hospital water have been positive since moving into the new building. Fifty-three (0.02%) of 219,521 patients had a positive Legionella test; 41 had pneumonia (40 community acquired). One definite L. pneumophila pneumonia confirmed by culture and DFA in August 1994 was nosocomial (0.0005%) by dates. This patient was transferred after prolonged hospitalization in another country, was transplanted 11 days after admission, and developed symptoms 5 days after liver transplant. However, tap water from the patient's room did not grow Legionella. Seventeen (2.5%) of 670 urine antigens were positive for Legionella (none nosocomial). Thirty-three (1.2%) of 2,671 DFAs were positive, including 7 patients (21%) without evidence of pneumonia and 6 (18%) who had an alternative diagnosis. CONCLUSION: Ultraviolet light usage was associated with negative water cultures and lack of clearly documented nosocomial Legionella infection for 13 years at this hospital.

A plasmid from a virulent strain of Legionella pneumophila is conjugative and confers resistance to ultraviolet light.

The presence of a single apparently cryptic plasmid of approximately 36 MDa was demonstrated in the virulent Dodge strain of Legionella pneumophila. 'Tagging' of the plasmid with Tn5 enabled transfer to be demonstrated to other strains of Legionella (though not to Escherichia coli or Pseudomonas aeruginosa) as well as a definitive assessment to be made of its stability. Plasmid carriage confers resistance to UV light probably by means of an error-prone UV repair system. The plasmid is compatible with plasmids of the IncP and IncW incompatibility groups.