Sewage Influent Research Articles

Determination of iodinated X-ray contrast media in sewage by solid-phase extraction and liquid chromatography tandem mass spectrometry

A method for the quantitative determination of five iodinated X-ray contrast media (ICMs) in sewage was developed by solid-phase extraction and high-performance liquid chromatography–tandem mass spectrometry. A fused-core analytical column was successfully applied for the first time for the separation of ICMs. Oasis HLB was selected from the sorbents tested because of its higher recoveries. The optimized method allowed the determination of the ICMs at low ng/L levels in both influent and effluent sewage, with detection limits of 40ng/L and 10ng/L for most compounds in influent and effluent sewage, respectively.The five ICMs studied were determined in all samples analysed, with iopromide being the analyte found at the highest concentration (8.9µg/L), while iopamidol was the analyte found at lowest concentration (1.3µg/L) in influent sewage. Effluent sewage did not show a significant decrease in ICM concentrations.

Experimental Studies of CS Type UV Sterilizer for the Urban Sewage Treatment

By treating the secondary effluent of Sanbaotun sewage treatment plant in Fushun with the CS type UV Sterilizer, the experiment researches the influence on the removal rate which includes the factors of turbidity, chromaticity, initial E.coli concentration of raw water, UV dose. The results show that the turbidity does not influence on UV disinfection efficiency with the turbidity from 1.5NTU to 3.8NTU. In the low-dose UV disinfection process, the influent sewage turbidity should under 4NTU to ensure the disinfection efficiency. Low chromaticity does not affect the UV disinfection efficiency, with the influent sewage chromaticity is under 15 degrees.The initial E. coli has a certain impact on little doses of UV disinfection with the concentration from 175 × 104 A/ L to 230 × 104 A/ L, no impact on high doses of UV disinfection。

Fate of stable strontium in the sewage treatment process as an analog for radiostrontium released by nuclear accidents

Radionuclides were widely released into the environment due to the nuclear accident at the Fukushima Daiichi Nuclear Power Plant. Some of these radionuclides have flowed into municipal sewage treatment plants through sewer systems. We have observed the fate of stable Sr in the sewage treatment process as a means to predict the fate of radiostrontium. Concentrations of stable Sr were determined in sewage influent, effluent, dewatered sludge, and incinerated sewage sludge ash collected from a sewage treatment plant once a month from July to December 2011. In the mass balance of Sr in the sewage treatment plant, 76% of the Sr entering the plant was discharged to the receiving water on average. Additionally, 14% of the Sr flowing through the plant was transferred to the sewage sludge and then concentrated in the sludge ash without being released to the atmosphere. We also investigated Sr sorption by activated sludge in a batch experiment. Measurements at 3 and 6h after the contact showed Sr was sorbed in the activated sludge; however, the measurements indicated Sr desorption from activated sludge occurred 48h after the contact.

Performance of UK wastewater treatment works with respect to trace contaminants

This study examined the performance of 16 wastewater treatment works to provide an overview of trace substance removal in relation to meeting the objectives of the Water Framework Directive (WFD). Collection and analysis of over 2400 samples including sewage influent, process samples at different stages in the treatment process and final effluent has provided data on the performance of current wastewater treatment processes and made it possible to evaluate the need for improved effluent quality. Results for 55 substances, including metals, industrial chemicals and pharmaceuticals are reported. Data for sanitary parameters are also provided. A wide range of removal efficiencies was observed. Removal was not clearly related to the generic process type, indicating that other operational factors tend to be important. Nonetheless, removals for many substances of current concern were high. Despite this, current proposals for stringent water quality standards mean that further improvements in effluent quality are likely to be required.

Molecular detection of Aichi virus in raw sewage in Italy

Aichi virus (AiV) is suspected to play a role in viral gastroenteritis in humans. In this study, we assessed the presence of AiV in untreated influent sewage samples collected at four wastewater treatment plants in central Italy. AiV was detected in 6 (12.5%) of the 48 specimens and in all plants. All of the Italian strains showed the highest nucleotide and amino acid sequence identity to genotype B AiV detected recently in Asia, especially in China.

A Microbial Signature Approach to Identify Fecal Pollution in the Waters Off an Urbanized Coast of Lake Michigan

Urban coasts receive watershed drainage from ecosystems that include highly developed lands with sewer and stormwater infrastructure. In these complex ecosystems, coastal waters are often contaminated with fecal pollution, where multiple delivery mechanisms that often contain multiple fecal sources make it difficult to mitigate the pollution. Here, we exploit bacterial community sequencing of the V6 and V6V4 hypervariable regions of the bacterial 16S rRNA gene to identify bacterial distributions that signal the presence of sewer, fecal, and human fecal pollution. The sequences classified to three sewer infrastructure-associated bacterial genera, Acinetobacter, Arcobacter, and Trichococcus, and five fecal-associated bacterial families, Bacteroidaceae, Porphyromonadaceae, Clostridiaceae, Lachnospiraceae, and Ruminococcaceae, served as signatures of sewer and fecal contamination, respectively. The human fecal signature was determined with the Bayesian source estimation program SourceTracker, which we applied to a set of 40 sewage influent samples collected in Milwaukee, WI, USA to identify operational taxonomic units (≥ 97 % identity) that were most likely of human fecal origin. During periods of dry weather, the magnitudes of all three signatures were relatively low in Milwaukee's urban rivers and harbor and nearly zero in Lake Michigan. However, the relative contribution of the sewer and fecal signature frequently increased to > 2 % of the measured surface water communities following sewer overflows. Also during combined sewer overflows, the ratio of the human fecal pollution signature to the fecal pollution signature in surface waters was generally close to that of sewage, but this ratio decreased dramatically during dry weather and rain events, suggesting that nonhuman fecal pollution was the dominant source during these weather-driven scenarios. The qPCR detection of two human fecal indicators, human Bacteroides and Lachno2, confirmed the urban fecal footprint in this ecosystem extends to at least 8 km offshore.

Sewage reflects the distribution of human faecal Lachnospiraceae

Faecal pollution contains a rich and diverse community of bacteria derived from animals and humans, many of which might serve as alternatives to the traditional enterococci and Escherichia coli faecal indicators. We used massively parallel sequencing (MPS) of the 16S rRNA gene to characterize microbial communities from wastewater treatment plant (WWTP) influent sewage from 12 cities geographically distributed across the USA. We examined members of the Clostridiales, which included the families Clostridiaceae, Lachnospiraceae and Ruminococcaceae for their potential as sewage indicators. Lachnospiraceae was one of the most abundant groups of faecal bacteria in sewage, and several Lachnospiraceae high-abundance sewage pyrotags occurred in at least 46 of 48 human faecal samples. Clone libraries targeting Clostridium coccoides (C. coccoides) in sewage samples demonstrated that Lachnospiraceae-annotated V6 pyrotags encompassed the previously reported C. coccoides group. We used oligotyping to profile the genus Blautia within Lachnospiraceae and found oligotypes comprised of 24 entropy components that showed patterns of host specificity. These findings suggest that indicators based on Blautia might have the capacity to discriminate between different faecal pollution sources. Development of source-specific alternative indicators would enhance water quality assessments, which leads to improved ecosystem health and reduced human health risk due to waterborne disease.

Comparison of the Microbial Community Structures of Untreated Wastewaters from Different Geographic Locales

Microbial sewage communities consist of a combination of human fecal microorganisms and nonfecal microorganisms, which may be residents of urban sewer infrastructure or flowthrough originating from gray water or rainwater inputs. Together, these different microorganism sources form an identifiable community structure that may serve as a signature for sewage discharges and as candidates for alternative indicators specific for human fecal pollution. However, the structure and variability of this community across geographic space remains uncharacterized. We used massively parallel 454 pyrosequencing of the V6 region in 16S rRNA genes to profile microbial communities from 13 untreated sewage influent samples collected from a wide range of geographic locations in the United States. We obtained a total of 380,175 high-quality sequences for sequence-based clustering, taxonomic analyses, and profile comparisons. The sewage profile included a discernible core human fecal signature made up of several abundant taxonomic groups within Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria. DNA sequences were also classified into fecal, sewage infrastructure (i.e., nonfecal), and transient groups based on data comparisons with fecal samples. Across all sewage samples, an estimated 12.1% of sequences were fecal in origin, while 81.4% were consistently associated with the sewage infrastructure. The composition of feces-derived operational taxonomic units remained congruent across all sewage samples regardless of geographic locale; however, the sewage infrastructure community composition varied among cities, with city latitude best explaining this variation. Together, these results suggest that untreated sewage microbial communities harbor a core group of fecal bacteria across geographically dispersed wastewater sewage lines and that ambient water quality indicators targeting these select core microorganisms may perform well across the United States.

Coxiella burnetii in sewage water at sewage water treatment plants in a Q fever epidemic area

During 2007-2010, over 4000 persons in The Netherlands contracted Q-fever, a zoonosis caused by the bacterium Coxiella burnetii. Goats and sheep are the main reservoir of C. burnetti and infected animals shed the bacterium with their urine, faeces and birth products. Human infections may occur through direct contact with infected animals, or through inhalation of contaminated dust particles or aerosols. Discharge of waste water from Q fever contaminated goat farms may result in the presence of C. burnetii in sewage water and aerosols at sewage water treatment plants (SWTPs) which may pose a health risk for workers or neighbouring residents. The objectives of this study were to determine the presence of C. burnetii at SWTPs and to optimize available detection methods. In March-July 2011, sewage influent and aeration tank samples from four SWTPs receiving discharge from Q fever positive goat farms were examined by using a multiplex real-time PCR detecting C. burnetii DNA by targeting IS1111 and com1 genes. Influent (44%; n=16/36) and active sludge (36%; n=13/36) samples were positive with low C. burnetii DNA content. Percentage positive samples per SWTP were 28-61%. Positive samples were most frequent in March 2011 and least frequent in May 2011. The presence of C. burnetii DNA in sewage water samples suggests that SWTPs receiving waste water from Q fever contaminated goat farms may contribute to the spread of C. burnetii to the environment. The low levels of C. burnetii DNA in sewage water during the decline of the Q fever outbreak in The Netherlands in 2011 indicate a low health risk for SWTP workers and residents.

Risk assessment for drugs of abuse in the Dutch watercycle

A screening campaign of drugs of abuse (DOA) and their relevant metabolites in the aqueous environment was performed in the Netherlands. The presence of DOA, together with the potential risks for the environment and the possible human exposure to these compounds through consumption of drinking water was investigated. Sewage water (influent and effluent), surface water of the rivers Rhine and Meuse, and drinking water (raw and finished) were analysed by four different laboratories using fully in-house validated methods for a total number of 34 DOA and metabolites. In this way, data reported for several compounds could also be confirmed by other laboratories, giving extra confidence to the results obtained in this study. In total 17 and 22 DOA were detected and quantified in influent and effluent sewage samples, respectively. The tranquilizers oxazepam and temazepam, and cocaine and its metabolite benzoylecgonine were found in high concentrations in sewage water. Nine compounds were possibly not efficiently removed during treatment and were detected in surface waters. The results indicated that substantial fractions of the total load of DOA and metabolites in the rivers Rhine and Meuse enter the Netherlands from abroad. For some compounds, loads appear to increase going downstream, which is caused by a contribution from Dutch sewage water effluents. As far as data are available, no environmental effects are expected of the measured DOA in surface waters.In raw water, three DOA were detected, whereas in only one finished drinking water out of the 17 tested, benzoylecgonine was identified, albeit at a concentration below the limit of quantification (<1 ng/L). Concentrations were well below the general signal value of 1 μg/L, which is specified for organic compounds of anthropogenic origin in the Dutch Drinking Water Act.

Performance of the linear ion trap Orbitrap mass analyzer for qualitative and quantitative analysis of drugs of abuse and relevant metabolites in sewage water

This work illustrates the potential of liquid chromatography coupled to a hybrid linear ion trap Fourier Transform Orbitrap mass spectrometer for the simultaneous identification and quantification of 24 drugs of abuse and relevant metabolites in sewage water. The developed methodology consisted of automatic solid-phase extraction using Oasis HLB cartridges, chromatographic separation of the targeted drugs, full-scan accurate mass data acquisition under positive electrospray ionization mode over an m/z range of 50–600Da at a resolution of 30,000 FWHM and simultaneous MSn measurements to obtain information of fragment ions generated in the linear ion trap. Accurate mass of the protonated molecule, together with at least one nominal mass product ion and retention time allowed the confident identification of the compounds detected in these complex matrices. In addition to the highly reliable qualitative analysis, Orbitrap analyzer also proved to have satisfactory potential for quantification at sub-ppb analyte levels, a possibility that has been very little explored in the literature until now. The limits of quantification ranged from 4 to 68ngL−1 in influent sewage water, and from 2 to 35ngL−1 in effluent, with the exception of MDA, morphine and THC that presented higher values as a consequence of the high ionization suppression in this type of samples. Satisfactory recoveries (70–120%) and precision (<30%) for the overall procedure were obtained for all compounds with the exception of meta-chlorophenylpiperazine, methylphenidate and ketamine. Isotope-labelled internal standards were added to sewage samples as surrogates in order to correct for matrix effects and also for possible losses during sample treatment. The methodology developed was applied to sewage water samples from the Netherlands (influent and effluent), and the results were compared with those obtained by LC–MS/MS with triple quadrupole. Several drugs of abuse could be identified and quantified, mainly MDMA, benzoylecgonine, codeine, oxazepam and temazepam. Orbitrap also showed potential for retrospective investigation of ketamine metabolites in the samples without the need of additional analysis.

Matrix effect in high‐performance liquid chromatography‐tandem mass spectrometry analysis of antibiotics in environmental water samples

This paper describes the matrix effect during the analysis of ten antibiotic compounds in water by SPE followed by HPLC-ESI-MS/MS. The target analytes were tetracycline, oxytetracycline (tetracyclines), sulfathiazole, sulfamethazine, sulfadiazine (sulfonamides), erythromycin-H(2)O, roxithromycine, spiramycin (macrolides), ofloxacin, and norfloxacin (quinolones). The matrix effect was examined for internal standards and the target analytes in five different water matrixes, with signal suppression being increased in the order: ultrapure water, tap water, river water, sewage effluent, and sewage influent. A combined application of the internal standards and matrix-matched extract calibration was shown to be successful in compensating the matrix effect for the analytes. The procedural recovery of the target compounds in sewage effluents and influents was higher than in river water samples, which was further enhanced by sample acidification to pH 2. The validity of the internal standard based matrix-matched calibration approach was verified by the standard addition method.

Occurrence and removal of pharmaceutical and hormone contaminants in rural wastewater treatment lagoons

Rural communities in the United States usually use a series of aerated lagoons to treat domestic wastewater. Effluents from these systems are typically discharged to receiving watersheds, which leads to a potential transfer of pharmaceuticals and personal care products (PPCPs) and steroid hormones from sanitary sewage to the environment. The primary objectives of this study are to identify and quantify PPCPs and steroid hormones in rural sewage treatment lagoons, to investigate the removal efficiency of these emerging contaminants in the treatment processes, and to monitor their occurrence in the surrounding watershed. In this study, a method has been developed to analyze thirteen PPCPs and eight steroid hormones in various water samples. Among all of the PPCPs considered, ten chemicals were detected in sewage influents, lagoon waters of different treatment stages, or effluents at concentrations in the ng/L to low μg/L range. Three hormones were observed in the influents at total concentrations as high as 164ng/L, but no hormone residues were detected in the effluents. This indicates that the aerated lagoons may effectively remove hormone contaminants. With the exception of carbamazepine, removal rates for the other detected PPCPs were relatively high in the range of 88 to 100% in September with average air temperature equal to 20°C. However, the removal efficiency of nine PPCPs in the rural wastewater treatment plant exhibited large temporal variability. The concentrations of PPCPs in the lagoon waters and effluents collected in November, with average air temperature equal to 4.4°C, were 1–2 orders of magnitude higher than those samples collected in September. Occurrence of these PPCP contaminants in the surrounding watershed was also monitored. The discharge of effluents significantly elevated the PPCP concentrations in the receiving creek and increased their occurrence in the adjacent river.

Extended Spectrum Beta-Lactamase Producing Bacteria in Waste Water Alexandria, Egypt

Waste water is a reservoir of resistant bacteria and an excellent location to describe the pattern of fecal carriage of extended-spectrum β lactamases (ESBLs) producing bacteria in the community. In this study, the aim was to determine the proportion of ESBL producing Gram-negative bacteria in waste water. Also to describe the antimicrobial susceptibility and types of beta-lactamases (TEM, SHV and CTX-M) among E. coli and K. pneumoniae isolates from waste water in the city of Alexandria, Egypt. The ESBLs compromised 69.8% in influent sewage and 57.7% in effluent sewage of the total Gram negative bacteria. The most frequently detected gene among E. coliisolates while blaTEM, while the most common among K. pneumonia isolates was blaSHV.

Determination of cyclic volatile methylsiloxanes in water, sediment, soil, biota, and biosolid using large-volume injection–gas chromatography–mass spectrometry

Several methods were developed to detect the cyclic volatile methylsiloxanes (cVMSs) including octamethylcyclotetrasiloxane (D4), decamethylcyclopentasiloxane (D5), and dodecamethylcyclohexasiloxane (D6) in water, sediment, soil, biota, and biosolid samples. Analytical techniques employed to optimize measurement of this compound class in various matrices included membrane-assisted solvent extraction in water, liquid–solid extraction for sediment, soil, biota, and biosolid samples. A subsequent analysis of the extract was conducted by large-volume injection–gas chromatography−mass spectrometry (LVI−GC−MS). These methods employed no evaporative techniques to avoid potential losses and contamination of the volatile siloxanes. To compensate for the inability to improve detection limits by concentrating final sample extract volumes we used a LVI–GC–MS. Contamination during analysis was minimized by using a septumless GC configuration to avoid cVMS’s associated with septum bleed. These methods performed well achieving good linearity, low limits of detection, good precision, recovery, and a wide dynamic range. In addition, stability of cVMS in water and sediment was assessed under various storage conditions. D4 and D5 in Type-I (Milli-Q) water stored at 4°C were stable within 29d; however, significant depletion of D6 (60–70%) occurred only after 3d. Whereas cVMS in sewage influent and effluent were stable at 4°C within 21d. cVMS in sediment sealed in amber glass jars at −20°C and in pentane extracts in vials at −15°C were stable during 1month under both storage conditions.

Investigation of degradation products of cocaine and benzoylecgonine in the aquatic environment

In this work, ultra-high-performance liquid chromatography (UHPLC) coupled to a hybrid quadrupole time-of-flight mass spectrometer (QTOF MS) has allowed the discovery and elucidation of degradation products of cocaine and its main metabolite benzoylecgonine (BE) in water. Spiked surface water was subjected to hydrolysis, chlorination and photo-degradation (both ultraviolet irradiation and simulated sunlight). After degradation of cocaine, up to sixteen compounds were detected and tentatively identified (1 resulting from hydrolysis; 8 from chlorination; 7 from photo-degradation), three of which are well known cocaine metabolites (BE, norbenzoylecgonine and norcocaine). Regarding BE degradation, up to ten compounds were found (3 from chlorination; 7 from photo-degradation), including one known metabolite (norbenzoylecgonine). Since reference standards were available for the major metabolites, they could be confirmed using information on retention time and fragment ions. The other degradates resulted from chlorination, dealkylation, hydroxylation and nitration, or from a combination of these processes. Several influent and effluent sewage water, and surface water samples were then screened for the identified compounds (known and unknown) using UHPLC–tandem MS with triple quadrupole. BE, norcocaine and norbenzoylecgonine were identified in these samples as major metabolites. Four previously unreported degradates were also found in some of the samples under study, illustrating the usefulness and applicability of the degradation experiments performed in this work.

Evaluation of two quantitative PCR assays using Bacteroidales and mitochondrial DNA markers for tracking dog fecal contamination in waterbodies

This study describes a comparative performance evaluation of two qPCR assays targeting a dog-associated Bacteroidales 16S rRNA genetic marker (CanBac-UCD) and a dog mitochondrial DNA (mtDNA) marker. The same fecal and environmental samples were assayed for the two markers thereby allowing direct comparison. A wide range of non-target species including, human, pig, horse, deer, mountain goat, bison, caribou, and moose were tested. Marker persistence was also monitored in freshwater microcosms. Both markers were prevalent in the canine samples collected in Regina, Saskatchewan and Calgary, Alberta, Canada (91% and 98% sensitivity, respectively). The mtDNA marker was detected exclusively in the target species while the CanBac-UCD marker was detected in all the non-target species (31% specificity). The CanBac-UCD marker exhibited faster decay in freshwater microcosms. The markers were rarely detected in the water samples collected from dog parks in Calgary and in Regina as well as from waterbodies and sewage influents in Saskatchewan, indicating possibly low to negligible levels of dog fecal contamination in the sampling areas. Altogether, the results of this study support the utility of the dog mtDNA assay in detecting dog fecal contamination in waterbodies.

Novel amide polar-embedded reversed-phase column for the fast liquid chromatography–tandem mass spectrometry method to determine polyether ionophores in environmental waters

A fast chromatographic method has been developed that takes less than 5min per run to determine five polyether ionophores with a novel amide polar-embedded reversed-phase column coupled to a triple quadrupole mass spectrometer. A comparison between Oasis HLB and Oasis MAX sorbents for the solid-phase extraction was done. Oasis HLB sorbent gave recoveries close to 90% and the repeatability (%RSD, 25–100ng/L, n=3) of the method was less than 7% for all compounds in all matrices. The presence of polyether ionophores in environmental waters such as river water and sewage was investigated. Monensin and narasin were frequently determined in influent and effluent sewage at concentrations from 10ng/L to 47ng/L in influents and from 6ng/L to 34ng/L in effluents. In river waters, polyether ionophores were not detected in any sample.

Improvement of Biofilm Carriers for the Treatment of Automotive Industry Wastewater

The efficient biological treatment of the wastewater of the automotive industry is difficult, mainly because of the low biological oxygen demand/chemical oxygen demand (BOD/COD) ratio. Moreover, wastewater of the automobile industry contains heavy metals and other chemical substances that may have toxic effects. Biofilm and hybrid technologies could be optimal solutions for the treatment of such heavily biodegradable wastewater streams. This paper presents the experimental results of the examination of modified biofilm carriers developed for the treatment of the wastewater of the automotive industry. Three modified biofilm carriers were examined: a high-density polyetyilene (HDPE), an HDPE modified with 50% carbon nanotube (CNT) content, and an HDPE modified with 75% CNT content. They were compared to a patented biofilm carrier having similar morphological properties (control). The examination consisted of two parts: (1) studying the colonisation of biofilm on each carrier, and (2) studying the biological COD removal efficiency of biofilm reactors filled with the four different biofilm carriers in 25% volumetric ratio each with an influent industrial emulsion sewage. The obtained results demonstrated that neither the rate of biofilm colonisation, nor the COD removal efficiency show any significant difference comparing the four biofilm carriers. The colonisation of biofilm was appropriate on each carrier and this ensured proper efficiency of COD removal in each biofilm reactor. Based on the results, it can be stated that the suggested advantageous characteristics of CNTs do not appear as it is mixed in the plastic raw material during the production of biofilm carriers. The absence of the significant difference observed between the examined carriers suggests that the production of biofilm carriers from recycled plastic could carry financial advantages compared to the control carrier. Further experiments could specify differences by various hydraulic loads or toxic effects.

Toxigenic Clostridium difficile PCR Ribotypes from Wastewater Treatment Plants in Southern Switzerland

The occurrence of Clostridium difficile in nine wastewater treatment plants in the Ticino Canton (southern Switzerland) was investigated. The samples were collected from raw sewage influents and from treated effluents. Forty-seven out of 55 characterized C. difficile strains belonged to 13 different reference PCR ribotypes (009, 010, 014, 015, 039, 052, 053, 066, 070, 078, 101, 106, and 117), whereas 8 strains did not match any of those available in our libraries. The most frequently isolated ribotype (40%) was 078, isolated from six wastewater treatment plants, whereas ribotype 066, a toxigenic emerging ribotype isolated from patients admitted to hospitals in Europe and Switzerland, was isolated from the outgoing effluent of one plant. The majority of the isolates (85%) were toxigenic. Forty-nine percent of them produced toxin A, toxin B, and the binary toxin (toxigenic profile A(+) B(+) CDT(+)), whereas 51% showed the profile A(+) B(+) CDT(-). Interestingly, eight ribotypes (010, 014, 015, 039, 066, 078, 101, and 106) were among the riboprofiles isolated from symptomatic patients admitted to the hospitals of the Ticino Canton in 2010. Despite the limitation of sampling, this study highlights that toxigenic ribotypes of C. difficile involved in human infections may occur in both incoming and outgoing biological wastewater treatment plants. Such a finding raises concern about the possible contamination of water bodies that receive wastewater treatment plant effluents and about the safe reuse of treated wastewater.