Photoreactivation of Legionella pneumophila after inactivation by low- or medium-pressure ultraviolet lamp

Photoreactivation of Legionella pneumophila after the inactivation by low-pressure (LP) or medium-pressure (MP) UV lamp was investigated in comparison with that of Escherichia coli. An endonuclease sensitive site (ESS) assay was used to determine the number of UV-induced pyrimidine dimers in the genome DNA of L. pneumophila or E. coli, while the survival ratio of each bacterium was also investigated by cultivation methods. L. pneumophila performed photoreactivation with almost complete repair of pyrimidine dimers associated with the quick recovery of survival ratio. A 3 log inactivation of L. pneumophila by LP or MP UV lamp was, respectively, resulted in 0.5 log or 0.4 log inactivation when photoreactivation was completed. Interestingly, L. pneumophila performed equivalent photoreactivation after LP and MP UV lamp exposures while photoreactivation of E. coli was significantly repressed after the inactivation by MP UV lamp. This study indicated that an attention would be required to design and operate a UV disinfection system targeting L. pneumophila. It was further implied that E. coli would not correctly indicate the fate of L. pneumophila in UV disinfection systems when photoreactivation takes place.     

The application of bioluminescence assay with culturing for evaluating quantitative disinfection performance

Various methods, including bioluminescence assay, were investigated regarding their suitability for quantitatively evaluating the disinfection performance. Although bioluminescence assay itself has been widely reported as a rapid, easy and suitable method for analyzing live microorganisms, the limited sensitivity of its measurement (approximately 10(3)-10(4)cells/assay vial), which is insufficient for disinfection study, requires further study. Among three methods (amplifying by enzymatic method, membrane filtration, and amplification by culturing) examined for increasing the detection sensitivity, amplification by culturing showed the best performance as Escherichia coli was employed as an indicating microorganism. Even with a short culturing time of 4h, the detection limit of E. coli measurement was successfully improved 200-fold, and the analytical results were not dependent upon the state of E. coli growth (stationary state with E. coli stock suspension vs. growth state with E. coli). In addition, the analytical integrity of bioluminescence assay with culturing was further demonstrated in comparison with spread plate method as free chlorine and UV irradiation were employed in the disinfection study.     

MBR出水的紫外线消毒试验研究

考察了紫外线（UV）对MBR出水中微生物的灭活情况，以及光活化和暗修复对细菌灭活效果的影响。结果表明：UV对MBR出水中的微生物具有良好的灭活效果，当UV剂量为16mJ／cm^2时，对细菌的对数灭活率为3-lg；在相同的UV剂量下，不同UV强度对细菌的灭活效果无显著影响；经UV消毒后的细菌在3h内未出现明显的暗修复现象，但在日光灯和太阳光辐射下可发生明显的光活化现象，且不同光源下的光活化速率和达到饱和的时间有所不同。     

紫外消毒出水的微生物光复活及其控制技术

污水厂出水经紫外线（UV）消毒后在排放过程中会出现微生物的复活现象，为此考察了采用UV-氯和UV-过氧乙酸（PAA）控制光复活的效果。经研究发现：在UV照射剂量为5．4mJ／cm^2、投氯量为2．5mg／L、反应时间为10min和UV照射剂量为5．4mJ／cm^2、过氧乙酸投量为10mg／L、反应时间为10min的条件下，对大肠菌群的灭活率均可达4个对数级以上，并能控制光复活现象。从消毒稳定性、经济适用性、安全毒副性等方面考虑，可采用UV—PAA作为污水厂出水消毒及抑制光复活的技术。     

Comparison of low- and medium-pressure ultraviolet lamps: Photoreactivation of Escherichia coli and total coliforms in secondary effluents of municipal wastewater treatment plants

It has been reported that Medium-Pressure (MP) ultraviolet (UV) lamps have an advantage over low-pressure (LP) lamps for water disinfection in terms of the photoreactivation of pure cultured bacteria. However, few studies have investigated the behavior of microorganisms in wastewater. Hence, in this study, the degree of photoreactivation, after UV exposure using both LP and MP lamps, in municipal wastewater samples was examined under a variety of conditions. Pure cultured Escherichia coli was also used to provide a comparison with previous studies.E. coli was found to undergo photoreactivation after both LP and MP exposure. The Colony Forming Ability (CFA) ratios were 0.60 and 0.32, and the percentage of photoreactivation was 50% and 20%, respectively, for LP and MP lamps with a germicidal UV dose of 5 mJ/cm². However, the advantage of the MP lamp was diminished for larger UV doses, since no photoreactivation was detected when the UV dose was 15 mJ/cm² for either LP or MP lamps. The microorganisms present in wastewater showed similar results to those of E. coli, however, no significant difference was found between the use of either a LP or a MP lamp. Also, when a UV dose of 40 mJ/cm² was applied, the percentage photoreactivation was less than 1%, no matter which type of lamp was used. From this work, it is concluded that the selection of the type of UV lamp for wastewater treatment plants, as regards photoreactivation of total coliforms, is not critical as long as the applied germicidal UV dose is greater than 40 mJ/cm².     

紫外线消毒对光复活时大肠杆菌活性的影响

以大肠杆菌为研究对象，考察了紫外线消毒对大肠杆菌的灭活效果及紫外线消毒后大肠杆菌在可见光下发生光复活的情况，同时考察了紫外线消毒和光复活过程中大肠杆菌电子传递体系（ETS）活性的变化。结果表明，大肠杆菌的灭活率随着紫外线剂量的增加而提高，在相同剂量下高强度的紫外线对大肠杆菌的灭活效果好于低强度的紫外线。经紫外线灭活后的大肠杆菌在可见光下会发生光复活，紫外线剂量对大肠杆菌的光复活有一定影响，高剂量下大肠杆菌发生光复活的能力比低剂量下的差；相同剂量下，较高强度的紫外线有利于控制大肠杆菌的光复活。对大肠杆菌ETS活性的研究表明，随着紫外线强度和剂量的增加，大肠杆菌的活性不断降低；紫外线消毒后的大肠杆菌经可见光照射后发生光复活时，其活性有所增加，在较高的紫外线剂量和强度下大肠杆菌的活性增加较少，而在较低剂量和强度下大肠杆菌的活性恢复能力较强。     

Sequential UV- and chlorine-based disinfection to mitigate Escherichia coli in drinking water biofilms

This study was designed to examine the potential downstream benefits of sequential disinfection to control the persistence of Escherichia coli under conditions relevant to drinking water distribution systems. Eight annular reactors (four polycarbonate and four cast iron) were setup in parallel to address various factors that could influence biofilm growth in distribution systems. Eight reactors were treated with chlorine, chlorine dioxide and monochloramine alone or in combination with UV to examine the effects on Escherichia coli growth and persistence in both the effluent and biofilm. In general, UV-treated systems in combination with chlorine or chlorine dioxide and monochloramine achieved greater log reductions in both effluent and biofilm than systems treated with chlorine-based disinfectants alone. However, during UV-low chlorine disinfection, E. coli was found to persist at low levels, suggesting that the UV treatment had instigated an adaptive mutation. During UV-chlorine-dioxide treatment, the E. coli that was initially below the detection limit reappeared during a low level of disinfection (0.2 mg/L) in the cast iron systems. Chloramine was shown to be effective in disinfecting suspended E. coli in the effluent but was unable to reduce biofilm counts to below the detection limit. Issues such as repair mechanism of E. coli and nitrification could help explain some of these aberrations. Improved understanding of the ability of chlorine-based disinfectant in combination with UV to provide sufficient disinfection will ultimately effect in improved management and safety of drinking water.     

紫外线和氯组合方式对大肠杆菌灭活效果的影响研究

系统地比较了紫外线和氯不同的组合方式对大肠杆菌的灭活效果.结果表明,紫外线和氯联合顺序消毒在一定程度上提高了消毒效果,但不同的组合方式以及两者不同的剂量对灭活效果都有影响.紫外线和氯先后作用的消毒效果优于紫外线和氯同时消毒.先氯后紫外线消毒时,加较高浓度的氯且短时间接触即可达到较好的消毒效果;先紫外线后氯消毒时,氯剂量越大则灭活率越高,若氯浓度相同而接触时间不同,则灭活效果差别不大.     

Ultraviolet and ionizing radiation for microorganism inactivation

The impacts of UV irradiation, gamma irradiation, and a combination of both on Escherichia coli inactivation in primary and secondary wastewater effluents were investigated. UV doses of 35 and 62 J/m(2) were required for a 1-log inactivation of E. coli in the primary and secondary wastewater samples, respectively. A gamma dose of 170 Gy (J/kg) was required for a 1-log inactivation of E. coli in both wastewater samples. Variation in gamma radiation dose rates did not have a significant impact on the extent of inactivation at a given total dose. Gamma irradiation of previously UV-irradiated samples indicated that particle-associated microorganisms, which are protected from UV, can be inactivated by ionizing radiation at a rate similar to that for free microorganism inactivation. An estimation of the energy required for disinfection indicated that, in general, the required energy and the energy cost for E. coli inactivation using ionizing radiation are considerably higher than those for UV radiation.     

Modelling of reactivation after UV disinfection: effect of UV-C dose on subsequent photoreactivation and dark repair

The increased use of UV radiation as a wastewater treatment technology has stimulated studies of the repair potential of microorganisms following treatment. In this study, samples of unfiltered secondary effluent were irradiated with seven levels of UV-C doses (50-200 mW s/cm(2)) from six low-pressure lamps in an open-channel UV disinfection system. Following irradiation, samples were incubated at 20 degrees C under photoreactivating light or in darkness. Samples were analysed for 240 min following incubation. The logistic model is proposed to explain the relation between photoreactivation and the UV-C dose received by the microorganisms. That model accurately fitted the data obtained in photoreactivation experiments, permitting interpretation of the estimated kinetic parameters: S(m) and k(2). In the experiments carried out in darkness, a slight reactivation is observed (<0.1%), followed by a decay period in which survival decreases. In order to model this last period, a modification was made to the logistic model by including a term of mortality that assumes a zero-order kinetic. The parameters S(m) and k(2), in both photoreactivation and darkness, show an exponential dependence on the UV-C inactivating dose. It is possible to predict their values, and hence the reactivation curve, from the equations proposed in this work.     

Inactivation/reactivation of antibiotic-resistant bacteria by a novel UVA/LED/TiO2 system

In this study, an effective photocatalytic disinfection system was established using the newly emerged high power UVA/LED lamp. Crystallizing dish coated with TiO₂ was prepared by 32-times impregnation-drying processes, and served as the supporting container for water samples. This study focused on the application of this UVA/LED system for the photocatalytic disinfection of selected antibiotic-resistant bacteria, Escherichia coli ATCC 700891. The disinfection performances were studied under various light intensities and illumination modes. Results show that higher light intensity could reach more significant inactivation of E. coli ATCC 700891. With the same UV dose, log-removal of antibiotic-resistant bacteria decreased with circle time in the studied range, while increased with duty circle. A “residual disinfecting effect” was found in the following dark period for bacteria collected at different phases of photocatalytic process. Residual disinfecting effect was found not significant for bacteria with 30 min periodic illumination. While residual disinfecting effect could kill almost all bacteria after 90 min UV periodic illumination within the following 240 min dark period.     

Inactivation of enteric microorganisms with chemical disinfectants, UV irradiation and combined chemical/UV treatments

The relative disinfection efficiencies of peracetic acid (PAA), hydrogen peroxide (H2O2) and sodium hypochlorite (NaOCl) against Escherichia coli, Enterococcus faecalis, Salmonella enteritidis and coliphage MS2 virus were studied in laboratory-scale experiments. This study also evaluated the efficiency of combined PAA/ultraviolet irradiation (UV) and H2O2/UV treatments to determine if the microbial inactivation was synergistic. Microbial cultures were added into a synthetic wastewater-like test medium and treated by chemical disinfectants with a 10 min contact time, UV irradiation or the combination of chemical and UV treatments. A peracetic acid dose of 3 mg/l resulted in approximately 2-3 log enteric bacterial reductions, whereas 7-15 mg/l PAA was needed to achieve 1-1.5 log coliphage MS2 reductions. Doses of 3-150 mg/l hydrogen peroxide achieved below 0.2 log microbial reductions. Sodium hypochlorite treatments caused 0.3-1 log microbial reductions at an 18 mg/l chlorine dose, while 2.6 log reductions of E. faecalis were achieved at a 12 mg/l chlorine dose. The results indicate that PAA could represent a good alternative to chlorine compounds in disinfection procedures, especially in wastewaters containing easily oxidizable organic matter. Hydrogen peroxide is not an efficient disinfectant against enteric microorganisms in wastewaters. The combined PAA/UV disinfection showed increased disinfection efficiency and synergistic benefits with all the enteric bacteria tested but lower synergies for the coliphage MS2. This suggests that this method could improve the efficiency and reliability of disinfection in wastewater treatment plants. The combined H2O2/UV disinfection only slightly influenced the microbial reductions compared to UV treatments and showed some antagonism and no synergies.     

Linear correlation between inactivation of E. coli and OH radical concentration in TiO2 photocatalytic disinfection

The biocidal action of the TiO2 photocatalyst has been now well recognized from massive experimental evidences, which demonstrates that the photocatalytic disinfection process could be technically feasible. However, the understanding on the photochemical mechanism of the biocidal action largely remains unclear. In particular, the identity of main acting photooxidants and their roles in the mechanism of killing microorganisms is under active investigation. It is generally accepted that reactive oxygen species (ROS) and OH radicals play the role. The aim of this study is to determine how the OH radical, acting either independently or in collaboration with other ROS, is quantitatively related to the inactivation of E. coli. The steady-state concentrations of OH radicals ([*OH]ss) in UV-illuminated TiO2 suspensions could be quantified from the measured photocatalytic degradation rates of p-chlorobenzoic acid (a probe compound) and its literature bimolecular rate constant with OH radicals. The results demonstrated an excellent linear correlation between [*OH]ss and the rates of E. coli inactivation, which indicates that the OH radical is the primary oxidant species responsible for inactivating E. coli in the UV/TiO2 process. The CT value of OH radical for achieving 2 log E. coli inactivation was initially found to be 0.8x10(-5) mg min/l, as predicted by the delayed Chick-Watson model. Although the primary role of OH radicals in photocatalytic disinfection processes has been frequently assumed, this is the first quantitative demonstration that the concentration of OH radicals and the biocidal activity is linearly correlated.     

Inactivation and potential repair of Cryptosporidium parvum following low- and medium-pressure ultraviolet irradiation

This study investigated the level of inactivation and the potential for Cryptosporidium parvum to repair following low doses (1 and 3mJ/cm(2)) of ultraviolet (UV) irradiation from both low- and medium-pressure UV lamps. Cryptosporidium parvum oocysts suspended in phosphate buffered saline were exposed to UV using a bench-scale collimated beam apparatus. Oocyst suspensions were incubated at 5 degrees C or 25 degrees C under light and dark conditions up to 120 h (5 days) following exposure to UV irradiation, to examine photoreactivation and dark repair potential, respectively. Cryptosporidium parvum infectivity was determined throughout the incubation period using an HCT-8 cell culture and an antibody staining procedure for detection. No detectable evidence of repair was observed after incubation under light or dark conditions following either LP or MP UV lamp irradiation.     

Impact of lamp shadowing and reflection on the fluence rate distribution in a multiple low-pressure UV lamp array

Use of mathematical modeling for determination of ultraviolet (UV) fluence in disinfection reactors requires accurate knowledge of the fluence rate distribution in a multiple lamp array. A method for measuring the fluence rate among a multiple lamp array was demonstrated using spherical actinometry. A matrix of four low-pressure UV lamps in air were investigated to evaluate the potential for shadowing and reflection to impact the fluence rate within and surrounding the lamp array. Two fluence rate distribution models were tested to determine the ability to predict the fluence rate distribution measured by the actinometers. Shadowing proved to attenuate UV light. Reflection from the lamp surface added 3-9% to the fluence rate, depending upon position in the reactor. These effects, as well as the fluence rate at various points in the lamp matrix were effectively modeled using RAD-LSI and UVCalc3D fluence rate distribution models. At fluence rates above 8mWcm(-2), the actinometry measured fluence rate was lower than the modeled rate, presumably from saturation of the actinometer solution at high fluence rates (close to the lamp). With multiple lamp reactors, the impact of shadowing can significantly affect fluence rate distribution and thus the level of microbial inactivation. If shadowing is not included in fluence rate distribution models, the fluence rate will be over predicted in the shadow zone of a neighboring lamp, falsely skewing model inactivation predictions.     

Application of a molecular biology concept for the detection of DNA damage and repair during UV disinfection

As nucleic acids are major targets in bacteria during standardised UV disinfection (254 nm), inactivation rates also depend on bacterial DNA repair. Due to UV-related DNA modifications, PCR-based approaches allow for a direct detection of DNA damage and repair during UV disinfection. By applying different primer sets, the correlation between amplicon length and PCR amplification became obvious. The longer the targeted DNA fragment was, the more UV-induced DNA lesions inhibited the PCR. Regeneration of Pseudomonas aeruginosa, Enterococcus faecium, and complex wastewater communities was recorded over a time period of 66 h. While phases of intensive repair and proliferation were found for P. aeruginosa, no DNA repair was detected by qPCR in E. faecium. Cultivation experiments verified these results. Despite high UV mediated inactivation rates original wastewater bacteria seem to express an enhanced robustness against irradiation. Regeneration of dominant and proliferation of low-abundant, probably UV-resistant species contributed to a strong post-irradiation recovery accompanied by a selection for β-Proteobacteria.     

UV-induced dark repair mechanisms in bacteria associated with drinking water

Caulobacter crescentus and Aquabacterium commune, both isolated from drinking water, as well as environmental isolates of Pseudomonas aeruginosa and Enterococcus faecium were treated with different UV fluences to study their capacity to restore induced DNA damages. Here, the induction of a key mechanism of bacterial dark repair, the so-called recA system, was analysed. With newly designed probes, the specific recA mRNA was detected by Northern blot. Additionally, the RecA protein was measured by the Western blot technique using a specific antibody. In drinking water bacteria as well as in opportunistic microorganisms, a specific induction of dark repair mechanisms was found even at UV fluences higher than 400J/m(2), the German standard for UV disinfection. This induction depended on the incubation time after UV treatment. Nevertheless, the UV-induced recA expressions were found to differ in the bacteria under investigation.     

Performance of UV disinfection and the microbial quality of greywater effluent along a reuse system for toilet flushing

This paper examines the microbial quality of treated RBC (Rotating Biological Contactor) and MBR (Membrane Bioreactor) light greywater along a continuous pilot-scale reuse system for toilet flushing, quantifies the efficiency of UV disinfection unit, and evaluates the regrowth potential of selected microorganisms along the system. The UV disinfection unit was found to be very efficient in reducing faecal coliforms and Staphylococcus aureus. On the other hand, its efficiency of inactivation of HPC (Heterotrophic Plate Count) and Pseudomonas aeruginosa was lower. Some regrowth occurred in the reuse system as a result of HPC regrowth which included opportunistic pathogens such as P. aeruginosa. Although the membrane (UF) of the MBR system removed all bacteria from the greywater, bacteria were observed in the reuse system due to “hopping phenomenon.” The microbial quality of the disinfected greywater was found to be equal or even better than the microbial quality of “clean” water in toilet bowls flushed with potable water (and used for excretion). Thus, the added health risk associated with reusing the UV-disinfected greywater for toilet flushing (regarding P. aeruginosa and S. aureus), was found to be insignificant. The UV disinfection unit totally removed (100%) the viral indicator (F-RNA phage, host: E. coli F_amp⁺) injected to the treatment systems simulating transient viral contamination. To conclude, this work contributes to better design of UV disinfection reactors and provides an insight into the long-term behavior of selected microorganisms along on-site greywater reuse systems for toilet flushing.     

Inactivation of bacteriophages in water by means of non-ionizing (UV-253.7 nm) and ionizing (gamma) radiation: a comparative approach

Thc inactivation behaviour of the bacteriophages PHI X 174 (ssDNA virus). MS2 (ssRNA virus) and B40-8 (dsDNA) toward non-ionizing (UV-253.7 nm) as well as to ionizing radiation (gamma radiation) was studied in order to evaluate their potential as viral indicators for water disinfection by irradiation. Previous findings of the high UV-253.7 nm resistance of MS2 were confirmed whereas an unexpected high sensitivity to gamma radiation compared to the two other phages was found. On the other hand, PHI X 174 revealed an enhanced UV sensitivity but a high resistance to ionizing radiation. B40-8 had an intermediate position between the other two bacteriophages relative to both types of radiation. As expected, the data of E. coli reconfirmed the unreliability of fecal indicator bacteria for the purpose of predicting responses of viruses to water treatment. In UV disinfection the influence of water matrix may be adequately controlled by considering the UV (253.7 nm) absorption of the water whereas so far no such parameter has existed for the influence of the water quality on ionizing irradiation with respect to the scavenger concentration.     

The effect of UV/H2O2 treatment on disinfection by-product formation potential under simulated distribution system conditions

Advanced oxidation with ultraviolet light and hydrogen peroxide (UV/H₂O₂) produces hydroxyl radicals that have the potential to degrade a wide-range of organic micro-pollutants in water. Yet, when this technology is used to reduce target contaminants, natural organic matter can be altered. This study evaluated disinfection by-product (DBP) precursor formation for UV/H₂O₂ while reducing trace organic contaminants in natural water (>90% for target pharmaceuticals, pesticides and taste and odor producing compounds and 80% atrazine degradation). A year-long UV/H₂O₂ pilot study was conducted to evaluate DBP precursor formation with varying water quality. The UV pilot reactors were operated to consistently achieve 80% atrazine degradation, allowing comparison of low pressure (LP) and medium pressure (MP) lamp technologies for DBP precursor formation. Two process waters of differing quality were used as pilot influent, i.e., before and after granular activated carbon adsorption. DBP precursors increased under most of the conditions studied. Regulated trihalomethane formation potential increased through the UV/H₂O₂ reactors from 20 to 118%, depending on temperature and water quality. When Post-GAC water served as reactor influent, less DBPs were produced in comparison to conventionally treated water. Haloacetic acid (HAA5) increased when conventionally treated water served as UV/H₂O₂ pilot influent, but only increased slightly (MP lamp) when GAC treated water served as pilot influent. No difference in 3-day simulated distribution system DBP concentration was observed between LP and MP UV reactors when 80% atrazine degradation was targeted.