Using total adenosine triphosphate (tATP) measurements for cyanobacterial bloom monitoring and response assessment during algaecide treatments.

Total adenosine triphosphate (tATP) was investigated for its potential as a rapid indicator of cyanobacterial growth and algaecide effectiveness. tATP and other common bloom monitoring parameters were measured over the growth cycles of cyanobacteria and green algae in laboratory cultures and examined at a drinking water source during an active bloom. Strong correlations (R2>0.78) were observed between tATP and chlorophyll-a in cyanobacteria cultures. tATP offered greater sensitivity by increasing two orders of magnitude approximately 7 d before changes in chlorophyll-a or optical density were observed in Lyngbya sp. and Dolichospermum sp. cultures. Increases in tATP per cell coincided with the onset of exponential growth phases in lab cultures and increase in cell abundance in field samples, suggesting that ATP/cell is a sensitive indicator that may be used to identify the development of blooms. Bench-scale trials using samples harvested during a bloom showed that tATP exhibited a clear dose-response during copper sulfate (CuSO4) and hydrogen peroxide (H2O2) treatment compared to chlorophyll-a and cell counts, indicating that cellular production and storage of ATP decreases even when live and dead cells cannot be distinguished. During Copper (Cu) algaecide application at a reservoir used as a drinking water source, tATP and cell counts decreased following initial algaecide application; however, the bloom rebounded within 10 d showing that the Cu algaecide only has limited effectiveness. In this case, tATP was a sensitive indicator to bloom rebounding after algaecide treatments and correlated positively with cell counts (R2=0.7). These results support the use of tATP as a valuable complementary bloom monitoring tool for drinking water utilities to implement during the monitoring and treatment of cyanobacterial blooms.

Critical Review on Bromate Formation during Ozonation and Control Options for Its Minimization.

Ozone is a commonly applied disinfectant and oxidant in drinking water and has more recently been implemented for enhanced municipal wastewater treatment for potable reuse and ecosystem protection. One drawback is the potential formation of bromate, a possible human carcinogen with a strict drinking water standard of 10 µg/L. The formation of bromate from bromide during ozonation is complex and involves reactions with both ozone and secondary oxidants formed from ozone decomposition, i.e., hydroxyl radical. The underlying mechanism has been elucidated over the past several decades, and the extent of many parallel reactions occurring with either ozone or hydroxyl radicals depends strongly on the concentration, type of dissolved organic matter (DOM), and carbonate. On the basis of mechanistic considerations, several approaches minimizing bromate formation during ozonation can be applied. Removal of bromate after ozonation is less feasible. We recommend that bromate control strategies be prioritized in the following order: (1) control bromide discharge at the source and ensure optimal ozone mass-transfer design to minimize bromate formation, (2) minimize bromate formation during ozonation by chemical control strategies, such as ammonium with or without chlorine addition or hydrogen peroxide addition, which interfere with specific bromate formation steps and/or mask bromide, (3) implement a pretreatment strategy to reduce bromide and/or DOM prior to ozonation, and (4) assess the suitability of ozonation altogether or utilize a downstream treatment process that may already be in place, such as reverse osmosis, for post-ozone bromate abatement. A one-size-fits-all approach to bromate control does not exist, and treatment objectives, such as disinfection and micropollutant abatement, must also be considered.

Design and operational considerations in response to Legionella occurrence in Las Vegas Valley groundwater.

Legionella occurrence monitoring is not required by United States Environmental Protection Agency (USEPA) drinking water regulations, and few occurrence studies exist for Legionella in source water or distribution systems. Legionella occurrence was monitored in Las Vegas Valley (Las Vegas, Nevada, USA) drinking water sources, including non-treated surface water, seasonal groundwater (61 wells, before and after chlorination), finished water (after treatment at water treatment facilities), and chlorinated distribution system water (at 9 reservoirs and 75 sample locations throughout the network). Legionella pneumophila was detected at least once at each of the wells sampled before chlorination, with an overall positivity rate of 38% (343/908). During well start-up (time<2 hours; turbidity>3 NTU), L. pneumophila concentrations averaged 2,792±353 MPN/100 mL, with a median of 105 MPN/100 mL, and range of <1 to 90,490 MPN/100 mL across 61 seasonally operated (typically April-October) groundwater wells. After initial flushing (turbidity<3 NTU), the average concentration decreased by more than two orders of magnitude to 24±3 MPN/100 mL but ranged from <1 to >2,273 MPN/100 mL. This trend indicates that stagnation (up to 391 days) contributed to greater initial concentrations, and flushing alone was incapable of complete L. pneumophila elimination. L. pneumophila concentration was significantly, positively correlated with total aqueous adenosine triphosphate (ATP) (p<0.00001, r=0.41-0.71), turbidity (p<0.00001, r=0.27-0.51), orthophosphate (p=0.35-0.076, r=0.51-0.59), and pump depth (p=0.032, r=0.40). During a full-scale assessment of chlorination (Ct=0.7 to 10.5 mg-min/L; T=26.6-28.1°C), substantial reduction of Legionella spp. (up to 2.5 logs) was observed; although, detectable concentrations were still measured. Extrapolating from a Chick-Watson model (log inactivation=0.28*(Ct); R2=0.87) constructed from the full-scale chlorination results, 3- and 4-log inactivation in Las Vegas Valley groundwater would require 10.8 and 14.3 mg-min/L, respectively; at least 3-log inactivation was required to bring Legionella spp. to below detection at the studied well. Chlorine exposure (Ct=0.1 to 10.9 mg-min/L) at most wells discharging directly to the distribution system was insufficient to fully inactivate Legionella spp. After discussing these findings with the state regulatory agency, direct-to-distribution wells (38 of 61 wells) remained out of operation; the distribution system, wells, and reservoirs were monitored for Legionella and chlorine residual, and additional treatment scenarios were identified for further evaluation. Legionella was either not detected or was well controlled in surface water, finished effluent from the drinking water treatment plant, chlorinated reservoirs, and the chlorinated distribution system. This study emphasizes the importance of utility-driven, non-regulatory research in order to protect public health and also identifies the need for greater occurrence monitoring and guidance for Legionella in groundwater supplies.

Ozone disinfection of waterborne pathogens and their surrogates: A critical review.

Viruses, Giardia cysts, and Cryptosporidium parvum oocysts are all major causes of waterborne diseases that can be uniquely challenging in terms of inactivation/removal during water and wastewater treatment and water reuse. Ozone is a strong disinfectant that has been both studied and utilized in water treatment for more than a century. Despite the wealth of data examining ozone disinfection, direct comparison of results from different studies is challenging due to the complexity of aqueous ozone chemistry and the variety of the applied approaches. In this systematic review, an analysis of the available ozone disinfection data for viruses, Giardia cysts, and C. parvum oocysts, along with their corresponding surrogates, was performed. It was based on studies implementing procedures which produce reliable and comparable datasets. Datasets were compiled and compared with the current USEPA Ct models for ozone. Additionally, the use of non-pathogenic surrogate organisms for prediction of pathogen inactivation during ozone disinfection was evaluated. Based on second-order inactivation rate constants, it was determined that the inactivation efficiency of ozone decreases in the following order: Viruses >> Giardia cysts > C. parvum oocysts. The USEPA Ct models were found to be accurate to conservative in predicting inactivation of C. parvum oocysts and viruses, respectively, however they overestimate inactivation of Giardia cysts at ozone Ct values greater than ∼1 mg min L-1. Common surrogates of these pathogens, such as MS2 bacteriophage and Bacillus subtilis spores, were found to exhibit different inactivation kinetics to mammalian viruses and C. parvum oocysts, respectively. The compilation of data highlights the need for further studies on disinfection kinetics and inactivation mechanisms by ozone to better fit inactivation models as well as for proper selection of surrogate organisms.

Benthic cyanobacteria: A utility-centred field study.

Although there is growing evidence that benthic cyanobacteria represent a significant source of toxins and taste and odour (T&O) compounds in water bodies globally, water utilities rarely monitor for them. Benthic cyanobacteria grow in an array of matrices such as sediments, biofilms, and floating mats, and they can detach and colonize treatment plants. The occurrence of compounds produced by benthic species across matrix and climate types has not been systematically investigated. Consequently, there is a lack of guidance available to utilities to monitor for and mitigate the risk associated with benthic cyanobacteria. To assess toxin and T&O risk across climatic zones and provide guidance to water utilities for the monitoring of benthic mats, two field surveys were conducted across three continents. The surveys examined the occurrence of six secondary metabolites and associated genes, namely, geosmin, 2-methylisoborneol (MIB), anatoxin-a, saxitoxin, microcystin, and cylindrospermopsin, in benthic environmental samples collected across three climates (i.e., temperate, sub-tropical, and tropical) and a range of matrix types. Existing enzyme-linked immunosorbent assays (ELISAs) and qPCR assays and were used to measure compound concentrations and their associated genes in samples. A novel qPCR assay was designed to differentiate the production of MIB by actinobacteria from that of cyanobacteria. MIB occurrence was higher in warmer climates than temperate climates. Cyanobacteria in benthic mats were the major producers of taste and odour compounds. Floating mats contained significantly higher concentrations of geosmin and saxitoxins compared to other matrix types. Samples collected in warmer areas contained significantly more saxitoxin and cylindrospermopsin than samples collected in temperate climates. While these trends were mainly indicative, they can be used to establish monitoring practices. These surveys demonstrate that benthic mats are significant contributors of secondary metabolites in source water and should be monitored accordingly. Benthic cyanobacteria were the sole producers of T&O in up to 17% of the collected samples compared to actinobacteria, which were sole producers in only 1% of the samples. The surveys also provided a platform of choice for the transfer of methodologies and specific knowledge to participating utilities to assist with the establishment of monitoring practices for benthic cyanobacteria and associated secondary metabolites.

A critical review on operation and performance of source water control strategies for cyanobacterial blooms: Part I-chemical control methods.

Cyanobacterial blooms produce nuisance metabolites (e.g., cyanotoxins and T&O compounds) thereby posing water quality management issues for aquatic sources used for potable water production, aquaculture, and recreation. A variety of in-lake/reservoir control measures are implemented to reduce the abundance of nuisance cyanobacteria biomass or decrease the amount of available phosphorous (P). This paper critically reviews the chemical control strategies implemented for in-lake/reservoir management of cyanobacterial blooms, i.e., algaecides and nutrient sequestering coagulants/flocculants, by highlighting (i) their mode of action, (ii) cases of successful and unsuccessful treatment, (iii) and factors influencing performance (e.g., water quality, process control techniques, source water characteristics, etc.). Algaecides generally result in immediate improvements in water quality and offer selective cyanobacterial control when peroxide-based alagecides are used. However, they have a range of limitations: causing cell lysis and release of cyanotoxins, posing negative impacts on aquatic plants and animals, leaving behind environmentally relevant treatment residuals (e.g., Cu in water and sediments), and offering only short-term bloom control characterized by cyanobacterial rebound. Coagulants/flocculants (alum, iron, calcium, and lanthanum bentonite) offer long-term internal nutrient control when external nutrient loading is controlled. Treatment performance is often influenced by background water quality conditions, and source water characteristics (e.g., surface area, depth, mixing regimes, and residence time). The reviewed case studies highlight that external nutrient load reduction is the most fundamental aspect of cyanobacterial control. None of the reviewed control strategies provide a comprehensive solution to cyanobacterial blooms.

A critical review on operation and performance of source water control strategies for cyanobacterial blooms: Part II-mechanical and biological control methods.

This review summarizes current knowledge on mechanical (artificial mixing, hypolimnetic aeration, dredging, and sonication) and biological (biomanipulation, macrophytes, and straws) methods for the management of cyanobacterial blooms in drinking water sources. Emphasis has been given to (i) the mechanism of cyanobacterial control, (ii) successful and unsuccessful case studies, and (iii) factors influencing successful implementation. Most mechanical and biological control strategies offer long-term control. However, their application can be cost-prohibitive and treatment efficacy is influenced by source water geometry and continual nutrient inputs from external sources. When artificial mixing and hypolimnetic oxygenation units are optimized based on source water characteristics, observed water quality benefits included increased dissolved oxygen contents, reduced internal loading of nutrients, and lower concentrations of reduced ions . Treatment efficacy during oxygenation and aeration was derailed by excessive sedimentation of organic matter and sediment characteristics such as low Fe/P ratios. Dredging is beneficial for contaminated sediment removal, but it is too costly to be a practical bloom control strategy for most systems. Sonication control methods have contradictory findings requiring further research to evaluate the efficacy and applicability for field-scale control of cyanobacteria. Biological control methods such as biomanipulation offer long-term treatment benefits; however, investigations on the mechanisms of field-scale cyanobacterial control are still limited, particularly with the use of macrophytes and straws. Each control method has site-specific strengths, limitations, and ecological impacts. Reduction of external nutrient inputs should still be a significant focus of restoration efforts as treatment benefits from mechanical and biological control were commonly offset by continued nutrient inputs.

Comparative Assessment of Physical and Chemical Cyanobacteria Cell Lysis Methods for Total Microcystin-LR Analysis.

Standardization and validation of alternative cell lysis methods used for quantifying total cyanotoxins is needed to improve laboratory response time goals for total cyanotoxin analysis. In this study, five cell lysis methods (i.e., probe sonication, microwave, freeze-thaw, chemical lysis with Abraxis QuikLyseTM, and chemical lysis with copper sulfate) were assessed using laboratory-cultured Microcystis aeruginosa (M. aeruginosa) cells. Methods were evaluated for destruction of cells (as determined by optical density of the sample) and recovery of total microcystin-LR (MC-LR) using three M. aeruginosa cell densities (i.e., 1 × 105 cells/mL (low-density), 1 × 106 cells/mL (medium-density), and 1 × 107 cells/mL (high-density)). Of the physical lysis methods, both freeze-thaw (1 to 5 cycles) and pulsed probe sonication (2 to 10 min) resulted in >80% destruction of cells and consistent (>80%) release and recovery of intracellular MC-LR. Microwave (3 to 5 min) did not demonstrate the same decrease in optical density (<50%), although it provided effective release and recovery of >80% intracellular MC-LR. Abraxis QuikLyseTM was similarly effective for intracellular MC-LR recovery across the different M. aeruginosa cell densities. Copper sulfate (up to 500 mg/L Cu2+) did not lyse cells nor release intracellular MC-LR within 20 min. None of the methods appeared to cause degradation of MC-LR. Probe sonication, microwave, and Abraxis QuikLyseTM served as rapid lysis methods (within minutes) with varying associated costs, while freeze-thaw provided a viable, low-cost alternative if time permits.

Delayed Release of Intracellular Microcystin Following Partial Oxidation of Cultured and Naturally Occurring Cyanobacteria.

Oxidation processes can provide an effective barrier to eliminate cyanotoxins by damaging cyanobacteria cell membranes, releasing intracellular cyanotoxins, and subsequently oxidizing these toxins (now in extracellular form) based on published reaction kinetics. In this work, cyanobacteria cells from two natural blooms (from the United States and Canada) and a laboratory-cultured Microcystis aeruginosa strain were treated with chlorine, monochloramine, chlorine dioxide, ozone, and potassium permanganate. The release of microcystin was measured immediately after oxidation (t ≤ 20 min), and following oxidant residual quenching (stagnation times = 96 or 168 h). Oxidant exposures (CT) were determined resulting in complete release of intracellular microcystin following chlorine (21 mg-min/L), chloramine (72 mg-min/L), chlorine dioxide (58 mg-min/L), ozone (4.1 mg-min/L), and permanganate (391 mg-min/L). Required oxidant exposures using indigenous cells were greater than lab-cultured Microcystis. Following partial oxidation of cells (oxidant exposures ≤ CT values cited above), additional intracellular microcystin and dissolved organic carbon (DOC) were released while the samples remained stagnant in the absence of an oxidant (>96 h after quenching). The delayed release of microcystin from partially oxidized cells has implications for drinking water treatment as these cells may be retained on a filter surface or in solids and continue to slowly release cyanotoxins and other metabolites into the finished water.

Using discrete and online ATP measurements to evaluate regrowth potential following ozonation and (non)biological drinking water treatment.

Water utilities must control microbial regrowth in the distribution system to protect public health. In this study, an adenosine triphosphate (ATP)-based biomass production potential test using indigenous bacterial communities were used to evaluate regrowth potential following ozonation with either biofiltration (BF) or sustained chlorination (SCl2). Two full-scale water treatment plants with different upstream processes (i.e., WTP-BF: ozonation, coagulation/flocculation, biofiltration, UV irradiation, chlorination; and WTP-SCl2: ozonation, chlorination, coagulation/flocculation, filtration, chlorination) were compared. Characterization of indigenous bacteria using 16S rRNA gene sequencing, qPCR, and cellular ATP (cATP) showed microbial diversity changes across treatment, biomass sloughing from biofilters (effluent cATP = 30 ±â€¯1 ng/L), and disinfection by chlorine (cATP < 1 ng/L). For both WTPs, 14-day cumulative biomass production (CBPt =  ∑t=0tATP(t)×Δt) was highest for ozonated water samples (CBP14 = 1.2 × 103-3.0 × 103 d ngATP/L). CBP further increased with increasing ozone dose due to production of more biodegradable carbon. Growth promotion by carbon was confirmed from the consumption of ozonation byproducts (carboxylic acids, aldehydes) and the increase in CBP (9.5 × 102-2.9 × 103 d ngATP/L) after addition of 50-300 µgC/L acetate. Ozone followed by sustained chlorination (WTP-SCl2) effectively controlled biomass growth across the treatment process (CBP14 <10 d ngATP/L). In contrast, ozone followed by biofiltration (WTP-BF) reduced regrowth potential by 30% (biofilter influent CBP14 = 1.3 × 103 d ngATP/L; biofilter effluent CBP14 = 9.3 × 102 d ngATP/L). After adding chlorine to the biofilter effluent, CBP14 was reduced to <10 d ngATP/L. Lastly, online ATP measurements confirmed the discrete measurements and improved identification of the cATP peak and growth phases of indigenous bacteria.

Using rapid quantification of adenosine triphosphate (ATP) as an indicator for early detection and treatment of cyanobacterial blooms.

Early detection of harmful cyanobacterial blooms allows identification of potential risk and appropriate selection of treatment techniques to prevent exposure in recreational water bodies and drinking water supplies. Here, luminescence-based adenosine triphosphate (ATP) analysis was applied to monitor and treat cultured and naturally occurring cyanobacteria cells. When evaluating lab-cultured Microcystis aeruginosa, ATP concentrations (≤252,000 pg/mL) had improved sensitivity and correlated well (R2 = 0.969) with optical density measurements at 730 nm (OD730; ≤0.297 cm-1). Following one year of monitoring of a surface water supply, ATP concentrations (≤2000 pg/mL) correlated (R2 = 0.791) with chlorophyll-a concentrations (≤50 µg/L). A preliminary early warning threshold of 175 pg ATP/mL corresponded with 5 µg/L chlorophyll-a to initiate increased monitoring (e.g., of cyanotoxins). Following oxidation processes (i.e., chlorine, chloramine, ozone, permanganate), ATP was demonstrated as an indicator of cell lysis and a threshold value of <100 pg/mL was recommended for complete release of intracellular cyanotoxins. ATP was also used to assess efficacy of copper (Cu(II)) treatment on cyanobacteria-laden surface water. While 24-h exposure to 2.5 mg Cu(II)/L did not impact chlorophyll-a, ATP decreased from 13,500 to 128 pg/mL indicating metabolic activity was minimized. Ultimately, ATP analysis holds promise for early detection and mitigation of potentially harmful algal blooms based on superior sensitivity, independence from cell morphology artifacts, rapid time for analysis (<10 min), and ease of deployment in the field compared to conventional methods.

Using upstream oxidants to minimize surface biofouling and improve hydraulic performance in GAC biofilters.

The combination of biological growth and particle loading can adversely affect hydraulic performance in drinking water biofilters. In this study, upstream oxidant addition was used to distribute biologically-derived filter clogging in granular activated carbon (GAC) biofilters. Oxidant penetration was assessed during pilot-scale operation and backwashing of dual media (GAC/sand) and multimedia (GAC/anthracite/sand) biofilters. Influent chlorine (HOCl), monochloramine (NH2Cl), and hydrogen peroxide (H2O2) residuals were optimized to react with the GAC surface in the upper portion of the filter media bed (depth < 0.5 m) to attenuate biomass development. As the oxidant residual was quenched by surface-mediated reaction with the filter media, biomass growth was promoted deeper in the filter bed (depth > 0.5 m). The oxidant-induced effects on biomass and hydraulic performance were monitored through measurements of adenosine triphosphate (ATP) and head loss accumulation at different media depths. Addition of oxidants (e.g., 0.6 mg Cl2/L HOCl) could decrease terminal head loss by 20% in dual media filters and 40% in multimedia filters. These hydraulic benefits were achieved without significantly affecting removal of assimilable organic carbon (AOC), total organic carbon (TOC), turbidity, and particle counts. Oxidant type, residual concentration, media type, media age, and media depth influenced the passage of oxidant residuals and distribution of filter biomass. When oxidants were added during backwashing, oxidant residual was quenched through the bed depth from a combination of reactions with GAC media and biofilm degradation. This attenuation of residual oxidant may prevent the oxidant residual from penetrating the entire bed depth, potentially compromising backwashing objectives.

Investigation of biotransformation, sorption, and desorption of multiple chemical contaminants in pilot-scale drinking water biofilters.

The evolving demands of drinking water treatment necessitate processes capable of removing a diverse suite of contaminants. Biofiltration can employ biotransformation and sorption to remove various classes of chemicals from water. Here, pilot-scale virgin anthracite-sand and previously used biological activated carbon (BAC)-sand dual media filters were operated for ∼250 days to assess removals of 0.4 mg/L ammonia as nitrogen, 50-140 µg/L manganese, and ∼100 ng/L each of trace organic compounds (TOrCs) spiked into pre-ozonated Colorado River water. Anthracite achieved complete nitrification within 200 days and started removing ibuprofen at 85 days. Limited manganese (10%) removal occurred. In contrast, BAC completely nitrified ammonia within 113 days, removed all manganese at 43 days, and exhibited steady state removal of most TOrCs by 140 days. However, during the first 140 days, removal of caffeine, DEET, gemfibrozil, naproxen, and trimethoprim decreased, suggesting a shift from sorption to biotransformation. Acetaminophen and sulfamethoxazole were removed at consistent levels, with complete removal of acetaminophen achieved throughout the study; ibuprofen removal increased with time. When subjected to elevated (1 µg/L) concentrations of TOrCs, BAC removed larger masses of chemicals; with a subsequent decrease and ultimate cease in the TOrCs spike, caffeine, DEET, gemfibrozil, and trimethoprim notably desorbed. By the end of operation, anthracite and BAC exhibited equivalent quantities of biomass measured as adenosine triphosphate, but BAC harbored greater microbial diversity (examined with 16S rRNA sequencing). Improved insight was gained regarding concurrent biotransformation, sorption, and desorption of multiple organic and inorganic contaminants in pilot-scale drinking water biofilters.

Microbial community characterization of ozone-biofiltration systems in drinking water and potable reuse applications.

Microbial community structure in the ozone-biofiltration systems of two drinking water and two wastewater treatment facilities was characterized using 16S rRNA gene sequencing. Collectively, these datasets enabled comparisons by facility, water type (drinking water, wastewater), pre-oxidation (ozonation, chlorination), media type (anthracite, activated carbon), media depth, and backwash dynamics. Proteobacteria was the most abundant phylum in drinking water filters, whereas Bacteroidetes, Chloroflexi, Firmicutes, and Planctomycetes were differentially abundant in wastewater filters. A positive correlation was observed between media depth and relative abundance of Cyanobacteria in drinking water filters, but there was only a slight increase in one alpha diversity metric with depth in the wastewater filters. Media type had a significant effect on beta but not alpha diversity in drinking water and wastewater filters. Pre-ozonation caused a significant decrease in alpha diversity in the wastewater filters, but the effect on beta diversity was not statistically significant. An evaluation of backwash dynamics resulted in two notable observations: (1) endosymbionts such as Neochlamydia and Legionella increased in relative abundance following backwashing and (2) nitrogen-fixing Bradyrhizobium dominated the microbial community in wastewater filters operated with infrequent backwashing. Bradyrhizobium is known to generate extracellular polymeric substances (EPS), which may adversely impact biofilter performance and effluent water quality. These findings have important implications for public health and the operation and resiliency of biofiltration systems.

Varied influence of microcystin structural difference on ELISA cross-reactivity and chlorination efficiency of congener mixtures.

Enzyme-linked immunosorbent assay (ELISA) is an antibody-based analytical method that has been widely applied in water treatment utilities for the screening of toxic cyanobacteria metabolites such as microcystins (MCs). However, it is unknown how the minor structural difference of MCs may impact their chlorination kinetics and measurement via ELISA method. It was found in this study that, regardless of the experimental conditions (n = 21), there was no MC-YR or MC-LY residual, while different removal rates of other MCs were observed (MC-RR > MC-LR > MC-LA âˆ¼ MC-LF) as measured by liquid chromatography tandem mass spectrometry (LC-MS/MS), which was consistent with the relative reactivity of the amino acid variables with free chlorine. The removal of total MCs was generally lower as measured by ELISA than by LC-MS/MS. By incorporating both analytical results, existence of ADDA-containing byproducts or byproducts that had a higher sensitivity toward the ELISA kit was demonstrated, after excluding the contribution of the cross-reactivity of the parent MCs. It should be noted, however, that the cross-reactivities of MCs could be influenced not only by MC congeners, but also by other conditions such as mixtures and the applied ELISA kit.

Ozone regeneration of granular activated carbon for trihalomethane control.

Spatial and temporal variations of trihalomethanes (THMs) in distribution systems have challenged water treatment facilities to comply with disinfection byproduct rules. In this study, granular activated carbon (GAC) and modified GAC (i.e., Ag-GAC and TiO2-GAC) were used to treat chlorinated tap water containing CHCl3 (15-21µg/L), CHBrCl2 (13-16µg/L), CHBr2Cl (13-14µg/L), and CHBr3 (3µg/L). Following breakthrough of dissolved organic carbon (DOC), GAC were regenerated using conventional and novel methods. GAC regeneration efficiency was assessed by measuring adsorptive (DOC, UV absorbance at 254nm, and THMs) and physical (surface area and pore volume) properties. Thermal regeneration resulted in a brief period of additional DOC adsorption (bed volume, BV, ∼6000), while ozone regeneration was ineffective regardless of the GAC type. THM adsorption was restored by either method (e.g., BV for ≥80% breakthrough, CHBr3 ∼44,000>CHBr2Cl ∼35,000>CHBrCl2 ∼31,000>CHCl3 ∼7000). Cellular and attached adenosine triphosphate measurements illustrated the antimicrobial effects of Ag-GAC, which may have allowed for the extended THM adsorption compared to the other GAC types. The results illustrate that ozone regeneration may be a viable in-situ alternative for the adsorption of THMs during localized treatment in drinking water distribution systems.

Colonial cell disaggregation and intracellular microcystin release following chlorination of naturally occurring Microcystis.

Colonial cell disaggregation and release of intracellular microcystin were evaluated following chlorine treatment of naturally occurring Microcystis. Microscopic observations of water samples collected from Lake Mead, Nevada, USA, confirmed the presence of colonial Microcystis with cells protected by an outer sheath up to 30 µm thick. During chlorination, two stages of cell decomposition were observed, stage 1: colonial cell disaggregation, and stage 2: unicellular degradation. Following a [Cl2]0:DOC0 ratio of 0.15 (t = 20 min, pH = 8.2-8.5) in unfiltered Lake Havasu samples, total particle count increased from (1.0 ± 0.11) × 10(5) to 4.2 × 10(5) particles/mL and fluorescent particle count increased from (1.2 ± 0.50) × 10(4) to 1.2 × 10(5) particles/mL, illustrating colonial cell disaggregation. Although total and fluorescent particles increased, the concentration of chlorophyll-a (Chl-a) decreased from 81 µg/L to 72 µg/L, and continued to decrease at higher [Cl2]0:DOC0 ratios. The preliminary second order rate constant for the reaction between Microcystis and chlorine in natural waters was estimated using either Chl-a (k = 15 M(-1) s(-1)) or fluorescence particle count (k = 38 M(-1) s(-1)) as an indicator of cell damage following colonial disaggregation (i.e., at [Cl2]0:DOC0 ratio ≥0.15). Complete release of intracellular microcystin-LR (MC-LR) was observed in both Lake Havasu and Lake Mead samples when applying a [Cl2]0:DOC0 ratio of 0.30 (t = 20 min), which was equivalent to a chlorine exposure of 8 min-mg/L for Lake Havasu samples. With chlorination, DOC increased by 3-18% indicating release of either colony-bound or cell-bound DOC. The results demonstrated the ability of chlorine to disaggregate/inactivate natural Microcystis colonies, and identified oxidation conditions resulting in complete release of intracellular MC-LR.

Evaluating fluorescence spectroscopy as a tool to characterize cyanobacteria intracellular organic matter upon simulated release and oxidation in natural water.

Intracellular organic matter (IOM) from cyanobacteria may be released into natural waters following cell death in aquatic ecosystems and during oxidation processes in drinking water treatment plants. Fluorescence spectroscopy was evaluated to identify the presence of IOM from three cyanobacteria species during simulated release into natural water and following oxidation processes (i.e. ozone, free chlorine, chloramine, chlorine dioxide). Peak picking and the fluorescence index (FI) were explored to determine which IOM components (e.g., pigments) provide unique and persistent fluorescence signatures with minimal interferences from the background dissolved organic matter (DOM) found in Colorado River water (CRW). When IOM was added to ultrapure water, the fluorescence signature of the three cyanobacteria species showed similarities to each other. Each IOM exhibited a strong protein-like fluorescence and fluorescence at Ex 370 nm and Em 460 nm (FDOM), where commercial fluorescence sensors monitor. All species also had strong phycobiliprotein fluorescence (i.e. phycocyanin or phycoerythrin) in the higher excitation range (500-650 nm). All three IOM isolates had FI values greater than 2. When IOM was added to CRW, phycobiliprotein fluorescence was quenched through interactions between IOM and CRW-DOM. Mixing IOM and CRW demonstrated that protein-like and FDOM intensity responses were not a simple superposition of the starting material intensities, indicating that interactions between IOM and CRW-DOM fluorescing moieties were important. Fluorescence intensity in all regions decreased with exposure to ozone, free chlorine, and chlorine dioxide, but the FI still indicated compositional differences compared to CRW-DOM. The phycobiliproteins in IOM are not promising as a surrogate for IOM release, because their fluorescence intensity is quenched by interactions with DOM and decreased during oxidation processes. Increases in both FDOM intensity and FI are viable qualitative indicators of IOM release in natural waters and following oxidation and may provide a more robust real-time indication of the presence of IOM than conventional dissolved organic carbon or UV absorbance measurements.

Effect of oxidant exposure on the release of intracellular microcystin, MIB, and geosmin from three cyanobacteria species.

The release of intracellular microcystin-LR (MC-LR), 2-methylisoborneol (MIB), and geosmin was investigated after the oxidation of three cyanobacteria (Microcystis aeruginosa (MA), Oscillatoria sp. (OSC), and Lyngbya sp. (LYN)). During the oxidation of 200,000 cells/mL of MA, release of intracellular MC-LR exceeded the World Health Organization (WHO) guideline of 1 µg/L during the lowest oxidant exposures (CT) tested: ozone (0 mg-min/L, below the ozone demand), chlorine (<40 mg-min/L), chlorine dioxide (<560 mg-min/L), and chloramine (<640 mg-min/L). As the CT increased, ozone, chlorine, and chlorine dioxide were able to oxidize the released MC-LR. During the oxidation of OSC (2800 cells/mL) and LYN (1600 cells/mL), release of intracellular MIB and geosmin exceeded reported threshold odor values after exposure to chlorine, chlorine dioxide, and chloramine, which have low reactivity with these taste and odor compounds. Ozone oxidation of OSC yielded an increase in MIB concentration at lower exposures (≤2.9 mg-min/L), likely due to insufficient oxidation by hydroxyl radicals. The release of intracellular organic matter (IOM) was also measured to determine the potential of bulk measurements to act as a surrogate for cyanotoxins and metabolite release. In all cases, the dissolved organic carbon (DOC) release was less than 0.25 mgC/L, which lacked the sensitivity to indicate the release of MC-LR, MIB, or geosmin. The fluorescence index proved to be a more sensitive indicator of intracellular organic matter release than DOC for MA. These results illustrate that toxic or odorous compounds may be released from cyanobacteria cells during oxidation processes with minimal changes in the DOC concentration.

Using digital flow cytometry to assess the degradation of three cyanobacteria species after oxidation processes.

Depending on drinking water treatment conditions, oxidation processes may result in the degradation of cyanobacteria cells causing the release of toxic metabolites (microcystin), odorous metabolites (MIB, geosmin), or disinfection byproduct precursors. In this study, a digital flow cytometer (FlowCAM(®)) in combination with chlorophyll-a analysis was used to evaluate the ability of ozone, chlorine, chlorine dioxide, and chloramine to damage or lyse cyanobacteria cells added to Colorado River water. Microcystis aeruginosa (MA), Oscillatoria sp. (OSC) and Lyngbya sp. (LYN) were selected for the study due to their occurrence in surface water supplies, metabolite production, and morphology. Results showed that cell damage was observed without complete lysis or fragmentation of the cell membrane under many of the conditions tested. During ozone and chlorine experiments, the unicellular MA was more susceptible to oxidation than the filamentous OSC and LYN. Rate constants were developed based on the loss of chlorophyll-a and oxidant exposure, which showed the oxidants degraded MA, OSC, and LYN according to the order of ozone > chlorine ~ chlorine dioxide > chloramine. Digital and binary images taken by the digital flow cytometer provided qualitative insight regarding cell damage. When applying this information, drinking water utilities can better understand the risk of cell damage or lysis during oxidation processes.