Approaching easy water disinfection for all: Can in situ electrochlorination outperform conventional chlorination under realistic conditions?

Electrochlorination has gained research interest for its potential application as decentralized water treatment. A number of studies have displayed promising efficiency for water disinfection. However, a comprehensive comparison of in situ electrodisinfection to existing disinfection techniques, particularly under realistic water composition and flow rates, still needs additional research efforts. The aim of this study is to evaluate in situ electrochlorination while comparing the treatment with conventional chemical chlorination for point-of-entry decentralized disinfection at the household level. An electrochemical flow cell reactor was operated in a single pass mode considering water flow and water consumption for a household of four family members. Disinfection efficiency assessment of both electrochemical and chemical chlorination was conducted using bacterial and viral surrogates, E. coli and MS2 bacteriophage. Furthermore, a techno-economic analysis was conducted, using the levelized cost of water, to compare two electrochemical chlorination scenarios (i.e., electrical grid energy use, and solar panel powered system) and benchmarked against the baseline treatment of chemical chlorination. The findings revealed increased inactivation efficiency of in situ electrochlorination over conventional chlorination (p-value < 0.05). The synergetic impact of radicals and chlorine, and/or contribution of high chlorine concentration at acidic pH near anode surface were identified as key factors that could enhance disinfection performance of in situ electrochlorination. The techno-economic analysis demonstrated that electrochemical treatment, when operated using renewable energy sources, is not only a more environmentally sustainable approach, but also emerges as a more economically feasible solution for decentralized water treatment application. The results highlight that in situ electrochlorination is a more advanced alternative to decentralized water chlorination. However, further fundamental research on products and by-products formation under various water matrices is required.

Impact of Pipe Material and Temperature on Drinking Water Microbiome and Prevalence of Legionella, Mycobacterium, and Pseudomonas Species.

In drinking water distribution systems (DWDSs), pipe material and water temperature are some of the critical factors affecting the microbial flora of water. Six model DWDSs consisting of three pipe materials (galvanized steel, copper, and PEX) were constructed. The temperature in three systems was maintained at 22 °C and the other 3 at 32 °C to study microbial and elemental contaminants in a 6-week survey using 16S rRNA next-generation sequencing (NGS) and inductively coupled plasma-optical emission spectrometry (ICP-OES). Pipe material and temperature were preferentially linked with the composition of trace elements and the microbiome of the DWDSs, respectively. Proteobacteria was the most dominant phylum across all water samples ranging from 60.9% to 91.1%. Species richness (alpha diversity) ranking was PEX < steel ≤ copper system and elevated temperature resulted in decreased alpha diversity. Legionellaceae were omni-prevalent, while Mycobacteriaceae were more prevalent at 32 °C (100% vs. 58.6%) and Pseudomonadaceae at 22 °C (53.3% vs. 62.9%). Heterogeneity between communities was disproportionately driven by the pipe material and water temperature. The elevated temperature resulted in well-defined microbial clusters (high pseudo-F index) in all systems, with the highest impact in PEX (10.928) followed by copper (9.696) and steel (5.448). Legionellaceae and Mycobacteriaceae are preferentially prevalent in warmer waters. The results suggest that the water temperature has a higher magnitude of impact on the microbiome than the pipe material.

Implication of cell culture methods and biases on UV inactivation of viruses.

Inactivation of human respiratory viruses in air and on surfaces is important to control their spread. Exposure to germicidal ultraviolet (UV-C) light damages viral nucleic acid rendering them non-infectious. Most of the recent viral inactivation studies have not considered potential artifacts caused by interactions between UV-C light and culture media used to suspend and deposit virus on surfaces. We show that the reactive oxygen and nitrogen species (ROS and RNS) form when commonly used virus culture media is exposed to 265 nm irradiation from light emitting diodes (LEDs) at UV-C doses (4 or 40 mJ/cm2) commonly considered to achieve multiple log-inactivation of virus. Surface viral inactivation values were enhanced from 0.49 to 2.92 log10 of viruses in DMEM, EMEM or EMEM-F as compared to absence of culture media (only suspended in Tris-buffer). The mechanisms responsible for the enhanced surface inactivate is hypothesized to involve photo-activation of vitamins and dyes present in the culture media, deposited with the virus on surfaces to be disinfected, which produce ROS and RNS. Given the rapidly growing research and commercial markets for UV-C disinfecting devices, there is a need to establish surface disinfecting protocols that avoid viral inactivation enhancement artifacts associated with selection and use of common cell culture media in the presence of UV-C light. This study addresses this weak link in the literature and highlights that inadequate selection of virus suspension media may cause a bias (i.e., over-estimation) for the UV-C dosages required for virus inactivation on surfaces.

Strain-Level Characterization of Legionella Environmental Isolates via MALDI-TOF-MS.

As a waterborne pathogen of increasing concern, techniques for cost-effective and rapid characterization of Legionella are vital. This study examines the development of a Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS) analysis methodology for this microbe. First, optimal sample preparation methods for the analysis of environmental Legionella isolates via MALDI-TOF-MS were determined. These methods were then implemented to perform strain-level characterization of environmental Legionella isolates from central Arizona. Results demonstrate that a MALDI-TOF-MS method involving BCYE agar-based culturing and protein extraction-based sample preparation yield high-quality mass spectra. Twenty-eight environmental Legionella isolates originating from two separate drinking water distribution systems were analyzed. Multiple species were detected, and strain-level characterization was achieved, with 12 unique strains distinguished. In addition, isolates of L. pneumophila, the most common species observed in the study, were correctly assigned to specific sampling sites. These results demonstrate the potential for this technique to be applied for sub-species characterization of Legionella with significant benefits over established methodologies.

Legionella Occurrence beyond Cooling Towers and Premise Plumbing.

Legionella is an environmental pathogen that is responsible for respiratory disease and is a common causative agent of water-related outbreaks. Due to their ability to survive in a broad range of environments, transmission of legionellosis is possible from a variety of sources. Unfortunately, a disproportionate amount of research that is devoted to studying the occurrence of Legionella in environmental reservoirs is aimed toward cooling towers and premise plumbing. As confirmed transmission of Legionella has been linked to many other sources, an over-emphasis on the most common sources may be detrimental to increasing understanding of the spread of legionellosis. This review aims to address this issue by cataloguing studies which have examined the occurrence of Legionella in less commonly investigated environments. By summarizing and discussing reports of Legionella in fresh water, ground water, saltwater, and distribution system drinking water, future environmental and public health researchers will have a resource to aid in investigating these pathogens in relevant sources.

Portable point-of-use photoelectrocatalytic device provides rapid water disinfection.

Portable water purification devices are needed to provide safe drinking water in rural communities, developing communities with low quality centralized water distribution, and military or recreational applications. Filtration, ultraviolet light, or chemical additives provide a spectrum of alternatives to remove pathogens from water. For the first time, we design, fabricate and demonstrate the performance of a small portable photoelectric point-of-use device, and document its performance on pathogen inactivation. The device utilizes a commercial teacup from which TiO2 nanotube photoanodes were produced in-situ and, with a small rechargeable battery powered 365 nm light emitting diode, was able to achieve 5-log inactivation of Escherichia coli in 10 s and 2.6-log of Legionella in 60 s of treatment in model water samples. Treatment of natural water achieved a 1-log bacteria inactivation after 30 s due to matrix effects. The electro-photocatalytic disinfection reactor in a kup (e-DRINK) can provide a feasible and affordable solution to ensure access to clean water. More broadly, this work demonstrates the potential for illumination to improve the efficiency of electrocatalytic surfaces.

Engineered proteoliposome transporter for treatment of cesium contaminated water.

Radioactive cesium (137Cs) released from nuclear power plants and nuclear accidents continues to be a worldwide concern, and its removal from water remains a difficult problem. Here, we present the development of an innovative method to remove Cs+ present at low concentrations in water. To achieve this, a proteoliposome transporter was engineered, composed of a membrane-bound potassium uptake protein, Kup from E. coli, which was reconstituted into a liposome vesicle. Cs+ removal (10-100 µg/L) was demonstrated by incubating the constructed proteoliposome in lab-fortified water, followed by ultracentrifugation to remove captured Cs+. Inductively coupled plasma mass spectrometry (ICP-MS) results from testing water spiked with 100 µg/L Cs+ revealed that adding increasing volumes of proteoliposome solution (containing 0.015-1.2 mg of Kup membrane transporter) resulted in 0.29-12.7% removal in a linear fashion. Proteoliposome addition (containing 0.015-0.3 mg of Kup membrane transporter) to water spiked with 10 µg/L Cs+ resulted in 0.65-3.43% removal, while removal by protein-free liposomes was negligible at 0.03%. These results suggest that Kup transporters inserted into the liposomes are mainly responsible for the removal efficiencies. Consequently, a desired removal efficiency can be achieved by adding a higher volume of constructed proteoliposome and subsequently higher mg of Kup transporter to the contaminated water. This provides new insight on the effectiveness and applicability of proteoliposome transporters, and an alternative and a novel contribution to emerging technologies in removing cesium or other metal contaminants undergoing transmembrane transport.

Nanoparticle and Transparent Polymer Coatings Enable UV-C Side-Emission Optical Fibers for Inactivation of Escherichia coli in Water.

Pathogenic bacteria pose a health threat and operational challenge in drinking water. UV-C light-emitting diodes (UV-C LEDs) are becoming a competitive disinfection technology but are limited by their small irradiation area. Side-emitting optical fibers (SEOFs) can serve as a UV-C LED light delivery technology for reactors or tubing. Modifying the surfaces of conventional optical fibers with scattering centers allows for side emission of 265 nm radiation from an LED for microbial inactivation in water. Solid-material absorbance and flux measurements differentiated light absorption from scattering for all materials. Silica spheres >200 nm in diameter achieved higher scattering than smaller silica. A critical discovery was that treating the silica-coated optical fiber in a solution of high ionic strength increased UV-C side emission by greater than 6-fold. Additionally, the cladding polymer Cytop had negligible absorbance at 265 nm wavelength. A scalable four-step treatment process was developed to fabricate the novel SEOF. Attached to a 265 nm LED, the side-emitting optical fiber achieved 2.9 log inactivation of Escherichia coli at a delivery dose of 15 mJ/cm2. The results illustrate proof of concept that UV-C SEOFs can inactivate E. coli and should be further explored for delivering LED light into water.

Physisorption and chemisorption of T4 bacteriophages on amino functionalized silica particles.

Bacteriophages, or phages, are receiving increasing interest as recognition tools for the design of bioactive surfaces. However, to maintain the activity of surface-bound phages, the immobilization strategy must provide the right orientation and not compromise the phages' integrity. The objectives of this study were to characterize the phage sorption capacity and the immobilized phage activity for aminated silica particles functionalized with T4 phages. Two functionalization strategies were compared; physisorption, based on electrostatic adhesion, and chemisorption, where the phage and the particle are coupled using a carbodiimide cross-linker. We report that chemisorption, at maximum adsorption conditions on 1 µm particles, yielded 16 functional phages per particle, which is 2.5 times more than by the physisorption method. Particle diameter is shown to have an important impact on phage attachment and 1.8 µm particles were found to have ∼4 times more phages per surface area than 0.5 µm particles. Higher surface coverage is attributed to the lower steric hindrance on bigger particles. These findings provide important guidelines for the design of phage-functionalized particles for environmental, biomedical, or sensing applications.

Copper nanoparticles toxicity: Laboratory strains verses environmental bacterial isolates.

Nanoparticles have emerged as significant environmental contaminants and their impact has been studied using laboratory strains of bacteria. This study focuses on investigating the response of environmental isolate and laboratory strains of E. coli to 50 and 100 nm size of copper nanoparticles (CuNPs). The laboratory cultures included pathogenic and non-pathogenic strains. The environmental isolate and the non-pathogenic E. coli strain showed different inactivation patterns. After 2 h exposure to 50 nm CuNPs, the environmental isolate and the lab strain of E. coli lost 7.22 and 6.47 log; whereas the reduction of 6.16 and 6.68 log resulted after exposure to 100 nm CuNPs, respectively. The pathogenic E. coli O157:H7 exposed to 50 and 100 nm CuNPs for 2 h resulted in 5.24 and 6.54 log reduction, respectively. Although the environmental isolate and the laboratory strains of E. coli showed similar inactivation trends; they exhibited different toxicity elicitation mechanisms after exposure to the CuNPs. The pathogenic and non-pathogenic strains elicited significantly different levels of glutathione reductase (GR) activities, an enzyme critical for protection against radicals. Similarly, the environmental isolate and the lab strains of E. coli exhibited opposite trend in GR activities. These results clearly indicate divergence in the toxicity elicitation in the environmental isolate versus the laboratory strains from exposure to CuNPs, which highlights the need for an in-depth investigation of the impact of NPs on the biological processes and long-term effect of high load of NPs on the stability of aquatic and terrestrial ecologies.

Legionella - A threat to groundwater: Pathogen transport in recharge basin.

This study elucidates the potential risk posed by Legionella during aquifer recharge practices. Experiments were conducted using pilot-scale column simulating infiltration of bacterial surrogate and pathogen, E. coli and Legionella pneumophila, under central Arizona recharge basin conditions. A column was packed with a loamy sand media collected from a recharge basin and was fitted with six sampling ports at soil depths of 15, 30, 60, 92, 122cm and acclimated for a month with tertiary treated wastewater. Transport of Legionella appeared to be delayed compared to E. coli. The breakthrough of E. coli and Legionella at 122cm depth occurred at 3 and 24h, respectively. Slow transport of Legionella is consistent with its pleomorphic nature and variation in size and shape under low nutrient conditions. Legionella persisted for a longer time in the column, but at lower concentrations. Given the novel results of this study, the transport of Legionella into groundwater aquifers can occur through engineering recharge basin conditions creating a potential public health risk.

A Strategy to Establish a Quality Assurance/Quality Control Plan for the Application of Biosensors for the Detection of E. coli in Water.

Rapid bacterial detection using biosensors is a novel approach for microbiological testing applications. Validation of such methods is an obstacle in the adoption of new bio-sensing technologies for water testing. Therefore, establishing a quality assurance and quality control (QA/QC) plan is essential to demonstrate accuracy and reliability of the biosensor method for the detection of E. coli in drinking water samples. In this study, different reagents and assay conditions including temperatures, holding time, E. coli strains and concentrations, dissolving agents, salinity and pH effects, quality of substrates of various suppliers of 4-methylumbelliferyl glucuronide (MUG), and environmental water samples were included in the QA/QC plan and used in the assay optimization and documentation. Furthermore, the procedural QA/QC for the monitoring of drinking water samples was established to validate the performance of the biosensor platform for the detection of E. coli using a culture-based standard technique. Implementing the developed QA/QC plan, the same level of precision and accuracy was achieved using both the standard and the biosensor methods. The established procedural QA/QC for the biosensor will provide a reliable tool for a near real-time monitoring of E. coli in drinking water samples to both industry and regulatory authorities.

A tool box strategy using Bacteroides genetic markers to differentiate human from non-human sources of fecal contamination in natural water.

Bacteroides genetic markers have been widely used to identify fecal pollution of water originating from human and animal sources. Many of the assays currently used for detecting human-specific Bacteroides produce false positive results. The focus of this study was to develop a microbial source tracking (MST) tool box strategy for differentiating Bacteroides from human and animal sources. Bacteroides 16S rRNA gene sequences from fish and selected animals were aligned against human fecal Bacteroides isolates to compare and characterize the variable regions within the 16S rRNA gene sequence. Conserved sequences between 4 variable regions were deleted and the truncated sequences were combined to develop a hyper-variable genomic segment (HVGS). The cladogram created from truncated sequences show a clear separation of Bacteroides from human feces and those from animal sources. The proposed strategy was field tested by collecting water samples from central Arizona source waters and three different recreational ponds. PCR using HF134 and HF183 primer sets was performed and sequences from positive reactions were aligned against human Bacteroides sequences to identify the source of contamination. Based on PCR results, the source of fecal contamination was presumptively identified as either human or from another source. For samples testing positive using the HF183 primer set (8/13), fecal contamination was presumed to be from human sources, but to confirm the results, PCR products were sequenced and aligned against the four variable regions and then incorporated within the truncated cladogram. As expected, the sequences from water samples with human fecal contamination grouped in a separate clade. A variability matrix, developed after exclusion of conserved sequences among the four regions, was utilized to establish discrete groupings for sequences within the truncated cladogram, generally differentiating Bacteroides isolates from varying host animals, but most importantly, separating Bacteroides from human feces from Bacteroides from other animals. The proposed strategy offers a new tool box method for MST and a step-wise methodology essential for identifying human sources of fecal pollution.

A biosensor platform for rapid detection of E. coli in drinking water.

There remains a need for rapid, specific and sensitive assays for the detection of bacterial indicators for water quality monitoring. In this study, a strategy for rapid detection of Escherichia coli in drinking water has been developed. This strategy is based on the use of the substrate 4-methylumbelliferyl-ß-d-glucuronide (MUG), which is hydrolyzed rapidly by the action of E. coli ß-d-glucuronidase (GUD) enzyme to yield a fluorogenic 4-methylumbelliferone (4-MU) product that can be quantified and related to the number of E. coli cells present in water samples. In this study, the detection time required for the biosensor response ranged between 20 and 120 min, depending on the number of bacteria in the sample. This approach does not need extensive sample processing with a rapid detection capability. The specificity of the MUG substrate was examined in both, pure cultures of non-target bacterial genera such as Klebsiella, Salmonella, Enterobacter and Bacillus. Non-target substrates that included 4-methylumbelliferyl-ß-d-galactopyranoside (MUGal) and l-leucine ß-naphthylamide aminopeptidase (LLß-N) were also investigated to identify nonspecific patterns of enzymatic activities in E. coli. GUD activity was found to be specific for E. coli and no further enzymatic activity was detected by other species. In addition, fluorescence assays were performed for the detection of E. coli to generate standard curves; and the sensitivity of the GUD enzymatic response was measured and repeatedly determined to be less than 10 E. coli cells in a reaction vial. The applicability of the method was tested by performing multiple fluorescence assays under pure and mixed bacterial flora in environmental samples. The results of this study showed that the fluorescence signals generated in samples using specific substrate molecules can be utilized to develop a bio-sensing platform for the detection of E. coli in drinking water. Furthermore, this system can be applied independently or in conjunction with other methods as a part of an array of biochemical assays in order to reliably detect E. coli in water.

The Impact of Capsid Proteins on Virus Removal and Inactivation During Water Treatment Processes.

This study examined the effect of the amino acid composition of protein capsids on virus inactivation using ultraviolet (UV) irradiation and titanium dioxide photocatalysis, and physical removal via enhanced coagulation using ferric chloride. Although genomic damage is likely more extensive than protein damage for viruses treated using UV, proteins are still substantially degraded. All amino acids demonstrated significant correlations with UV susceptibility. The hydroxyl radicals produced during photocatalysis are considered nonspecific, but they likely cause greater overall damage to virus capsid proteins relative to the genome. Oxidizing chemicals, including hydroxyl radicals, preferentially degrade amino acids over nucleotides, and the amino acid tyrosine appears to strongly influence virus inactivation. Capsid composition did not correlate strongly to virus removal during physicochemical treatment, nor did virus size. Isoelectric point may play a role in virus removal, but additional factors are likely to contribute.

Impact of environmental factors on legionella populations in drinking water.

To examine the impact of environmental factors on Legionella in drinking water distribution systems, the growth and survival of Legionella under various conditions was studied. When incubated in tap water at 4 °C, 25 °C, and 32 °C, L. pneumophila survival trends varied amongst the temperatures, with the stable populations maintained for months at 25 °C and 32 °C demonstrating that survival is possible at these temperatures for extended periods in oligotrophic conditions. After inoculating coupons of PVC, copper, brass, and cast iron, L. pneumophila colonized biofilms formed on each within days to a similar extent, with the exception of cast iron, which contained 1-log less Legionella after 90 days. L. pneumophila spiked in a model drinking water distribution system colonized the system within days. Chlorination of the system had a greater effect on biofilm-associated Legionella concentrations, with populations returning to pre-chlorination levels within six weeks. Biofilms sampled from drinking water meters collected from two areas within central Arizona were analyzed via PCR for the presence of Legionella. Occurrence in only one area indicates that environmental differences in water distribution systems may have an impact on the survival of Legionella. These results document the impact of different environmental conditions on the survival of Legionella in water.

Automobile windshield washer fluid: A potential source of transmission for Legionella.

Epidemiological evidence suggesting driving cars to be a risk factor for legionellosis has prompted public health studies to investigate vehicle windshield washer fluid as a novel transmission source of this disease. The goal of the current study was to investigate whether or not windshield washer fluid could serve as a potential source of transmission for Legionella. A wide variation in the survival of L. pneumophila was observed when incubated in different washer fluids at 25 and 37 °C, however, one brand tested supported Legionella survival similar to or greater than sterilized deionized water. In addition, 1 L of tap water contained in a washer fluid reservoir was able to support population growth and survival of Legionella for several months. In a field study examining the windshield washer fluid of 12 elementary school buses, Legionella were detected from 84% of samples at a high concentration of 8.1×10(4) CFU/mL. Culturable cells were also detected in aerosolized washer fluid during washer fluid spray. By demonstrating survival in certain windshield washer fluids, growth within washer fluid reservoirs, and the presence of viable cells in bus washer fluid spray, we have provided evidence suggesting the potential for a novel route of Legionella exposure.

Occurrence of Noroviruses and Their Correlation with Microbial Indicators in Raw Milk.

This study was conducted to investigate the microbiological quality of raw cow's milk in a collection center in the city of Mashhad, Iran. A total of 19 raw cow's milk samples were collected and simultaneously analyzed for male-specific (F(+)) coliphage and Escherichia coli using culture-based methods and for enteric viruses by reverse transcriptase semi-nested PCR using primer sets specific for human norovirus Group I (HNV-GI), human norovirus Group II (HNV-GII), and enteroviruses (EV). Seven out of 19 (36.8%) raw milk samples tested positive for human noroviruses (HNV). The genotypes detected were HNV-GI and HNV-GII. Three positive samples contained both genotypes, and 2 samples were positive for either of HNV-GI and HNV-GII. No sample tested positive for EV. The correlation between the occurrence of HNV and the microbial indicators was studied. The statistical analysis using first- and second-order regression revealed that there is no correlation between F(+) coliphage and E. coli. Similarly, no correlation was noticed between the occurrence of F(+) coliphages and HNV. However, frequency distribution analysis indicated that 3 out of 4 (75%) of raw milk samples containing F(+) coliphage at a concentration higher than 10(4) pfu/100 ml were also positive for noroviruses. The limited data on the occurrence of noroviruses in raw milk suggest a poor sanitation and hygiene practices at the facility and indicate a possible correlation between the viral indicator at high concentration and human noroviruses; however, this analysis needs further investigation in a larger scale study.

Stimulation of 2-methylisoborneol (MIB) production by actinomycetes after cyclic chlorination in drinking water distribution systems.

The impact of fluctuation in chlorine residual on actinomycetes and the production of 2-methylisoborneol (MIB) were studied in cast-iron and PVC model distribution systems. Actinomycetes were spiked in each system and continued operation for a 12-day non-chlorine experiment, resulting in no changes in actinomycetes and MIB concentrations. Three cyclic chlorination events were performed and chlorine residuals were maintained as follows: 1.0 mg L(-1) for 24 h, 0 mg L(-1) for 48 h, 0.5 mg L(-1) for 48 h, 0 mg L(-1) for 48 h and 2 mg L(-1) for 24 h. After each chlorination event, 2 -3 log decrease in actinomycetes was noted in both systems. However, within 48 h at 0 mg L(-1) chlorine, the actinomycetes recovered to the pre-chlorination levels. On the contrary, MIB concentration in both systems remained un-impacted after the first cycle and increased by fourfold (< 5 to > 20 mg L(-1)) after the second cycle, which lasted through the third cycle despite the fact that actinomycetes numbers fluctuated 2-3 logs during this time period. For obtaining biofilm samples from field, water meters were collected from municipality drinking water distribution systems located in central Arizona. The actinomycetes concentration in asbestos cement pipe and cast iron pipe averaged 3.1 × 10(3) and 1.9 × 10(4) CFU cm(-2), respectively. The study shows that production of MIB is associated with changes in chlorine residual in the systems. This is the first report of cyclic chlorine shock as a stimulus for MIB production by actinomycetes in drinking water distribution system's ecology.

Evaluation of the relationship between bulk organic precursors and disinfection byproduct formation for advanced oxidation processes.

Advanced oxidation processes (AOPs) are gaining traction as they offer mineralization potential rather than transferring contaminants between media. However, AOPs operated with limited energy and/or chemical inputs can exacerbate disinfection byproduct (DBP) formation, even as precursors such as dissolved organic carbon, UV254, and specific UV absorbance (SUVA) decrease. This study examined the relationship between DBP precursors and formation using TiO2 photocatalysis experiments, external AOP and non-AOP data, and predictive DBP models. The top-performing indicator, SUVA, generally correlated positively with trihalomethanes and haloacetic acids, but limited-energy photocatalysis yielded contrasting negative correlations. The accuracy of predicted DBP values from models based on bulk parameters was generally poor, regardless of use and extent of AOP treatment and type of source water. Though performance improved for scenarios bounded by conditions used in model development, only 0.5% of the model/dataset pairings satisfied all measured parameter boundary conditions, thereby introducing skepticism toward model usefulness. Study findings suggest that caution should be employed when using bulk indicators and/or models as a metric for AOP mitigation of DBP formation potential, particularly for limited-energy/chemical inputs.