Framework to Quantify Uncertainty in Microplastic Concentrations in Wastewaters and Sludges Incorporating Analytical Recovery Information into Data Analysis.

Wastewater treatment plants (WWTPs) serve a pivotal role in transferring microplastics (MPs) from wastewater to sludge streams, thereby exerting a significant influence on their release into the environment and establishing wastewater and biosolids as vectors for MP transport and delivery. Hence, an accurate understanding of the fate and transport of MPs in WWTPs is vital. Enumeration is commonly used to estimate concentrations of MPs in performance evaluations of treatment processes, and risk assessment also typically involves MP enumeration. However, achieving high accuracy in concentration estimates is challenging due to inherent uncertainty in the analytical workflow to collect and process samples and count MPs. Here, sources of random error in MP enumeration in wastewater and other matrices were investigated using a modeling approach that addresses the sources of error associated with each step of the analysis. In particular, losses are reflected in data analysis rather than merely being measured as a validation step for MP extraction methods. A model for addressing uncertainty in the enumeration of microorganisms in water was adapted to include key assumptions relevant to the enumeration of MPs in wastewater. Critically, analytical recovery, the capacity to successfully enumerate particles considering losses and counting error, may be variable among MPs due to differences in size, shape, and type (differential analytical recovery) in addition to random variability between samples (nonconstant analytical recovery). Accordingly, differential analytical recovery among the categories of MPs was added to the existing model. This model was illustratively applied to estimate MP concentrations from simulated data and quantify uncertainty in the resulting estimates. Increasing the number of replicates, counting categories of MPs separately, and accounting for both differential and nonconstant analytical recovery improved the accuracy of MP enumeration. This work contributes to developing guidelines for analytical procedures quantifying MPs in diverse types of samples and provides a framework for enhanced interpretation of enumeration data, thereby facilitating the collection of more accurate and reliable MP data in environmental studies.

A comprehensive floc model for simulating simultaneous nitrification, denitrification, and phosphorus removal.

A comprehensive floc model for simultaneous nitrification, denitrification, and phosphorus removal (SNDPR) was designed, incorporating polyphosphate-accumulating organisms (PAOs), glycogen-accumulating organisms (GAOs), intrinsic half-saturation coefficients, and explicit external mass transfer terms. The calibrated model was able to effectively describe experimental data over a range of operating conditions. The estimated intrinsic half-saturation coefficients of oxygen values for ammonia-oxidizing bacteria, nitrite-oxidizing bacteria, ordinary heterotrophic organisms (OHOs), PAOs, and GAOs were set at 0.08, 0.18, 0.03, 0.07, and 0.1 mg/L, respectively. Simulation suggested that low dissolved oxygen (DO) environments favor K-strategist nitrifying bacteria and PAOs. In SNDPR, virtually all influent and fermentation-generated volatile fatty acids were assimilated as polyhydroxyalkanoates by PAOs in the anaerobic phase. In the aerobic phase, PAOs absorbed 997 % and 171 % of the benchmark influent total phosphorus mass loading through aerobic growth and denitrification via nitrite. These high percentages were because they were calculated relative to the influent total phosphorus, rather than total phosphorus at the end of the anaerobic period. When considering simultaneous nitrification and denitrification, about 23.1 % of influent total Kjeldahl nitrogen was eliminated through denitrification by PAOs and OHOs via nitrite, which reduced the need for both oxygen and carbon in nitrogen removal. Moreover, the microbial and DO profiles within the floc indicated a distinct stratification, with decreasing DO and OHOs, and increasing PAOs towards the inner layer. This study demonstrates a successful floc model that can be used to investigate and design SNDPR for scientific and practical purposes.

Quantifying the removal of polybrominated diphenyl ethers (PBDEs) in physical, chemical, and biological sludge treatment systems.

Polybrominated diphenyl ethers (PBDE) are priority contaminants historically used as flame retardants. PBDEs are known to occur in wastewater biosolids posing potential concerns with the beneficial land application of the biosolids. This study evaluated the removal of 21 congeners in nine full-scale sludge treatment systems including pelletization (P), alkaline stabilization (AS), and aerobic (AE) and anaerobic (AN) digestion. It is the first study to conduct a mass balance analysis of a broad spectrum of PBDEs during physical, chemical, and biological sludge treatment. The PBDE congener pattern in raw sludge and biosolids samples was consistent with commercial formulations. The fully brominated congener BDE-209 dominated biosolids from all sites with an average concentration of 620 ng/g dry weight (dw), followed by BDE-99 (173 ng/g dw) and BDE-47 (162 ng/g dw). Mass balance analysis on the P and AS processes showed no change in PBDE mass flows with treatment. However, aerobic and anaerobic digestion processes reported significant levels of removal and formation of individual congeners, though the results were not consistent between facilities. One aerobic digestion process (AE2) reported an overall average removal of 48%, whereas the other (AE1) reported very high levels of accumulation of tri- and tetraBDE congeners. Similarly, there were significant variations in PBDE behavior across the five anaerobic digestion plants studied. The plant with the longest solids retention time (SRT) (AN1) reported a moderate removal (50%) of overall PBDE loading and lower congeners, whereas other plants (AN2-AN5) showed significant low (-19%) to high (-166%) levels of formation of lower congeners. The results suggest that reduced SRTs result in formation of lower congeners while extended SRTs can lead to moderate removal of some PBDEs. Conventional sludge treatment result in low to moderate PBDE removal and advanced thermal conversion technologies may be needed to improve the contaminant removal during sludge treatment.

Implementing an adaptive, two-tiered SARS-CoV-2 wastewater surveillance program on a university campus using passive sampling.

Building-level wastewater-based surveillance (WBS) has been increasingly applied upstream from wastewater treatment plants to conduct targeted monitoring for SARS-CoV-2. In this study, a two-tiered, trigger-based wastewater surveillance program was developed on a university campus to monitor dormitory wastewater. The objective was to determine if passive sampling with cotton gauze as a sampling medium could be used to support institution-level public health action. Two nucleocapsid gene targets (N1 and N2) of SARS-CoV-2 as well as the endogenous fecal indicator pepper mild mottle virus (PMMoV) were quantified using RT-qPCR. >500 samples were analyzed during two contrasting surveillance periods. In the Fall of 2021 community viral burden was low and a tiered sampling network was able to isolate individual clinical cases at the building-scale. In the Winter of 2022 wastewater signals were quickly elevated by the emergence of the highly transmissible SARS-CoV-2 Omicron (B.1.1.529) variant. Prevalence of SARS-CoV-2 shifted surveillance objectives from isolating cases to monitoring trends, revealing both the benefits and limitations of a tiered surveillance design under different public health situations. Normalization of SARS-CoV-2 by PMMoV was not reflective of upstream population differences, suggesting saturation of the material occurred during the exposure period. The passive sampling method detected nearly all known clinical cases and in one instance was able to identify one pre-symptomatic individual days prior to confirmation by clinical test. Comparisons between campus samplers and municipal wastewater influent suggests that the spread of COVID-19 on the campus was similar to that of the broader community. The results demonstrate that passive sampling is an effective tool that can produce semi-quantitative data capable of tracking temporal trends to guide targeted public health decision-making at an institutional level. Practitioners of WBS can utilize these results to inform surveillance program designs that prioritize efficient resource use and rapid reporting.

Phosphorus release and recovery by reductive dissolution of chemically precipitated phosphorus from simulated wastewater.

Chemically mediated recovery of phosphorous (P) as vivianite from the sludges generated by chemical phosphorus removal (CPR) is a potential means of enhancing sustainability of wastewater treatment. This study marks an initial attempt to explore direct P release and recovery from lab synthetic Fe-P sludge via reductive dissolution using ascorbic acid (AA) under acidic conditions. The effects of AA/Fe molar ratio, age of Fe-P sludge and pH were examined to find the optimum conditions for Fe-P reductive solubilization and vivianite precipitation. The performance of the reductive, chelating, and acidic effects of AA toward Fe-P sludge were evaluated by comparison with hydroxylamine (reducing agent), oxalic acid (chelating agent), and inorganic acids (pH effect) including HNO3, HCl, and H2SO4. Full solubilization of Fe-P sludge and reduction of Fe3+ were observed at pH values 3 and 4 for two Fe/AA molar ratios of 1:2 and 1:4. Sludge age (up to 11 days) did not affect the reductive solubilization of Fe-P with AA addition. The reductive dissolution of Fe-P sludge with hydroxylamine was negligible, while both P (95 ± 2%) and Fe3+ (90 ± 1%) were solubilized through non-reductive dissolution by oxalic acid treatment at an Fe/oxalic acid molar ratio 1:2 and a pH 3. With sludge treatment with inorganic acids at pH 3, P and Fe release was very low (<10%) compared to AA and oxalic acid treatment. After full solubilization of Fe-P sludge by AA treatment at pH 3 it was possible to recover the phosphorus and iron as vivianite by simple pH adjustment to pH 7; P and Fe recoveries of 88 ± 2% and 90 ± 1% respectively were achieved in this manner. XRD analysis, Fe/P molar ratio measurements, and magnetic attraction confirmed vivianite formation. PHREEQC modeling showed a reasonable agreement with the measured release of P and Fe from Fe-P sludge and vivianite formation.

ß-Cyclodextrin functionalized adsorbents for removal of organic micropollutants from water.

The presence of organic micropollutants in water is an ongoing concern due to the potential risks to living organisms. ß-Cyclodextrin-based adsorbents have been developed to remove organic micropollutants from water as they are deemed to be efficient, selective and reusable. This literature review establishes the current state of the knowledge on the application of ß-Cyclodextrin adsorbents for the removal of organic micropollutants from water and determines knowledge gaps and recommendations for future studies. An inventory of organic micropollutants that have been studied was developed and it revealed that bisphenol-A has been the most commonly studied. Adsorbent configurations were reviewed and modifications to the adsorbent structures that have provided enhanced adsorption properties were identified. The size and shape of the organic micropollutants was found to affect the adsorption behavior. The surface charge of ß-Cyclodextrin adsorbents influence adsorption when repulsive forces are present and the extent of repulsion can depend on the pH of the solution. Common competitors such as natural organic matter and inorganic ions do not significantly impact the adsorption of organic micropollutants however relatively small fulvic acids may compete for the ß-Cyclodextrin cavity depending on the adsorbent type. Desorption of organic micropollutants from these adsorbents has been accomplished with alcohols and most adsorbents have been recovered and reused in adsorption/desorption cycles. The need for enhanced recovery processes that maintain water quality and adsorbent integrity was identified. The use of quantitative structure-activity relationships and molecular computational tools could potentially guide future environmental applications of ß-Cyclodextrin adsorbents.

Nitrogen removal pathways during simultaneous nitrification, denitrification, and phosphorus removal under low temperature and dissolved oxygen conditions.

Nitrogen removal pathways of simultaneous nitrification, denitrification, and phosphorus removal (SNDPR) at low dissolved oxygen (0.3 mg/L) and temperature (10℃) were explored to understand nitrogen removal mechanisms. Biological nitrogen and phosphorus removal was sustained with total inorganic nitrogen removal, phosphorus removal, and simultaneous nitrification and denitrification (SND) efficiencies of 62.6%, 97.3%, and 31.2%, respectively. The SND was observed in the first 2 h of the aerobic phase and was attributed to denitrifying ordinary heterotrophic organisms using readily biodegradable chemical oxygen demand and denitrifying phosphorus accumulating organisms (DPAOs), which removed 15.1% and 12.2% of influent nitrogen, respectively. A phosphorus accumulating organism (PAO)-rich community was indicated by stoichiometric ratios and supported by 16S rRNA gene analysis, with Dechloromonas, Zoogloea, and Paracoccus as DPAOs, and Ca. Accumulibacter and Tetrasphaera as PAOs. Even though Ca. Competibacter (10.4%) was detected, limited denitrifying glycogen accumulating organism denitrification was observed.

Removal and formation of perfluoroalkyl substances in Canadian sludge treatment systems - A mass balance approach.

Poly- and per-fluoroalkyl substances (PFAS) are an emerging class of anthropogenic contaminants whose occurrence has raised concerns with the beneficial reuse of biosolids from wastewater treatment. This study evaluated the behavior of thirteen PFAS in nine Canadian sludge treatment systems including pelletization, alkaline stabilization, aerobic and anaerobic digestion processes. The composition of the overall PFAS-fluorine (ΣPFAS-F) loading in a system fed with only primary sludge was dominated by perfluorodecanoate (PFDA), whereas systems with blended primary and waste activated sludge feeds had a mix of short and long chain PFAS in raw sludges and treated biosolids. An increase in average ΣPFAS-F mass flow was observed through pelletization (19% formation) and alkaline stabilization (99% formation) processes indicating negative removal or contaminant formation. One of the two aerobic digestion systems and three of the five anaerobic digestion systems showed modest reductions (< 40% removal) in ΣPFAS-F loading. Long chain PFAS such as perfluorodecanoate (PFDA) and perfluorooctane sulfonate (PFOS) exhibited a wide variation in behavior ranging from substantial formation (> 75% formation) to modest removal (42% removal) in the surveyed systems while short chain perfluoropentanoate (PFPeA) mass flows increased through the three systems where they occurred. Overall, the contaminant mass balances revealed that there were significant changes in mass flows of the target PFAS through all kinds of sludge treatment systems. The results of this study on PFAS fate through sludge processing can inform future global PFAS risk management activities as well as sludge treatment considerations.

Organic phosphorus removal using an integrated advanced oxidation-ultrafiltration process.

Non-reactive phosphorus (nRP) contains condensed phosphates and organic phosphorus (OP) species that are recalcitrant in secondary wastewater treatment and tend to remain in final effluents. To meet ultra-low effluent P discharge limits, persistent nRP must be removed. The objective of this study was to evaluate the use of an advanced oxidation process (AOP) which couples TiO2/UV photolysis with ultrafiltration to oxidize and remove nRP species. Initial tests utilized OP model compounds, adenosine triphosphate (ATP) and aminoethylphosphonate (AEP), in a binary mixture to evaluate AOP treatment and to elucidate possible mechanisms of phosphorus removal. The results were consistent with a model of preferential ATP binding to the TiO2 surface compared to AEP. On UV light exposure, AEP was removed from solution due to the photooxidation of ATP freeing up binding sites for AEP adsorption and subsequent oxidation. Orthophosphate released during AOP treatment was retained on the TiO2 solids. The AOP was applied to three municipal wastewaters and one automotive industry effluent for P removal. In all cases, phosphorus removal was found to occur through filtration, surface complexation and UV oxidation. Total phosphorus removal efficiencies between 90 and 97% were observed for the municipal wastewater effluents and 44% removal was observed in the industrial effluent after treatment using AOP.

Characterization of performance of full-scale tertiary membranes under stressed operating conditions.

This study sought to identify factors responsible for enhanced fouling of ultrafiltration membranes used in tertiary wastewater treatment under challenging conditions of high flow and low temperature. A detailed analysis of full-scale membrane operating data was conducted, and this was supported by data gathered through a field sampling campaign. Higher average fouling rates and average recoveries were observed during periods of highest flows and lowest temperatures. The results demonstrated that the negative impact of seasonal changes on short-term fouling are readily reversible, while hydraulically irreversible fouling, which is responsible for intermediate and long-term fouling rates, is not effectively recovered by maintenance cleans (MCs) but is recovered by recovery cleans (RCs). An examination of membrane feedwater quality revealed that high fouling rates correlated to an increase in dissolved organic carbon (DOC) concentrations, with the biopolymer fraction of the DOC being most important. Increased capillary suction time (CST) values, which indicate reduced sludge dewaterability, were also observed during high fouling events. It was concluded that seasonal variations result in the increased release of extracellular polymeric substances (EPS) by microorganisms, which leads to higher membrane fouling and worsened dewaterability of the activated sludge.

High functional diversity among Nitrospira populations that dominate rotating biological contactor microbial communities in a municipal wastewater treatment plant.

Nitrification, the oxidation of ammonia to nitrate via nitrite, is an important process in municipal wastewater treatment plants (WWTPs). Members of the Nitrospira genus that contribute to complete ammonia oxidation (comammox) have only recently been discovered and their relevance to engineered water treatment systems is poorly understood. This study investigated distributions of Nitrospira, ammonia-oxidizing archaea (AOA), and ammonia-oxidizing bacteria (AOB) in biofilm samples collected from tertiary rotating biological contactors (RBCs) of a municipal WWTP in Guelph, Ontario, Canada. Using quantitative PCR (qPCR), 16S rRNA gene sequencing, and metagenomics, our results demonstrate that Nitrospira species strongly dominate RBC biofilm samples and that comammox Nitrospira outnumber all other nitrifiers. Genome bins recovered from assembled metagenomes reveal multiple populations of comammox Nitrospira with distinct spatial and temporal distributions, including several taxa that are distinct from previously characterized Nitrospira members. Diverse functional profiles imply a high level of niche heterogeneity among comammox Nitrospira, in contrast to the sole detected AOA representative that was previously cultivated and characterized from the same RBC biofilm. Our metagenome bins also reveal two cyanase-encoding populations of comammox Nitrospira, suggesting an ability to degrade cyanate, which has only been shown previously for several Nitrospira representatives that are strict nitrite oxidizers. This study demonstrates the importance of RBCs as model systems for continued investigation of environmental factors that control the distributions and activities of AOB, AOA, comammox Nitrospira, and other nitrite oxidizers.

Impact of anaerobically digested biosolids characteristics and handling conditions on dewatering performance at multiple facilities.

Anaerobically digested biosolids (ABD) characteristics that affect dewatering were assessed at three water resource recovery facilities (WRRF) with different handling practices. Dewatering performance at the three sites corresponded to different levels of soluble chemical oxygen demand (COD), ammonia (NH4 -N), and mono- and divalent cation concentrations in ADB. Capillary suction time (CST) and a modified centrifugal technique were used to determine optimum polymer doses and to assess the impact of handling conditions on dewatering performance. Both techniques indicated that polymer dosing between 15 and 20 kg/dry tonne was optimal for all facilities and that biosolids mixing and pumping did not significantly impact dewaterability. The CST values of anaerobically digested biosolids decreased as temperature increased, but no significant difference was found for either temperature or location of dewatering facilities. Sludge viscosity and rheological properties that vary with temperature appeared to have influenced CST values. Modified centrifugal technique results indicated cake solids were not affected by polymer make-up water or ADB temperature when emulsion polymer was used. This study shows the value of laboratory testing of biosolids under controlled conditions to identify and correct potential problems in full-scale operations. PRACTITIONER POINTS: Capillary suction time and a modified centrifugal technique were used to assess the impact of different process-related and environmental factors on dewatering. Higher concentrations of soluble COD (potentially extracellular polymeric substances - EPS) and low calcium (Ca) in anaerobically digested biosolids align with reduced dewaterability. Cell disruption and break down of floc structures due to storage/mixing and pumping of biosolids did not appear to negatively impact dewatering. Modified centrifugal test results did not provide conclusive evidence of whether dewatering of anaerobically digested biosolids could be significantly impacted by temperature over the range 15-30°C, especially when emulsion polymer is used. This study shows the value of laboratory testing of biosolids under controlled conditions to identify potential problems in the full-scale operations.

Effect of solids residence time on dynamic responses in chemical P removal.

The impact of solids residence time (SRT) on the dynamics of phosphorus (P) removal by hydrous ferric oxide (HFO) floc was characterized through experimental and modeling studies. Three abiotic process conditions were considered in systems operated over a range of SRTs (~3 to 27 days): uptake in sequencing batch reactors (SBRs) under (a) constant and (b) dynamic P loading conditions, and (c) uptake in batch sorption tests with preformed HFO solids. P removal under all conditions was characterized by an initial period of fast removal followed by a period of slower removal until pseudo-equilibrium was reached. The initial removal rate increased with increasing P concentrations and was attributed to a larger concentration gradient between soluble- and adsorbed-phase concentrations. A kinetic model was developed and found to describe the dynamic behavior of P adsorption onto HFO floc under all conditions tested. A consistent mass transfer rate coefficient (k) was found to describe mass transfer over a range of SRTs for low initial P concentrations. At elevated SRTs (23-27 days) and elevated influent P concentrations, k values were found to deviate from those estimated at reduced SRTs. Differences in process mixing conditions were reflected in the estimated rate coefficients (k). Integration of the kinetic model with existing equilibrium models in wastewater process simulators will improve the ability to predict P uptake onto HFO floc under dynamic loading conditions in water resource recovery facilities. Models that consider the kinetics of P uptake will be particularly relevant for facilities that are required to achieve ultralow P concentrations. PRACTITIONER POINTS: This work provides a kinetic model that can be integrated with existing equilibrium models in wastewater process simulators to improve the ability to predict P uptake onto HFO floc under dynamic loading conditions. This research can be used to assist WRRFs to achieve ultralow effluent P requirements.

Transcriptomics investigation of thyroid hormone disruption in the olfactory system of the Rana [Lithobates] catesbeiana tadpole.

Thyroid hormones (THs) regulate vertebrate growth, development, and metabolism. Despite their importance, there is a need for effective detection of TH-disruption by endocrine disrupting chemicals (EDCs). The frog olfactory system substantially remodels during TH-dependent metamorphosis and the objective of the present study is to examine olfactory system gene expression for TH biomarkers that can evaluate the biological effects of complex mixtures such as municipal wastewater. We first examine classic TH-response gene transcripts using reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) in the olfactory epithelium (OE) and olfactory bulb (OB) of premetamorphic Rana (Lithobates) catesbeiana tadpoles after 48 h exposure to biologically-relevant concentrations of the THs, 3,5,3'-triiodothyronine (T3) and L-thyroxine (T4), or 17-beta estradiol (E2); a hormone that can crosstalk with THs. As the OE was particularly sensitive to THs, further RNA-seq analysis found >30,000 TH-responsive contigs. In contrast, E2 affected 267 contigs of which only 57 overlapped with THs suggesting that E2 has limited effect on the OE at this developmental phase. Gene ontology enrichment analyses identified sensory perception and nucleoside diphosphate phosphorylation as the top affected terms for THs and E2, respectively. Using classic and additional RNA-seq-derived TH-response gene transcripts, we queried TH-disrupting activity in municipal wastewater effluent from two different treatment systems: anaerobic membrane bioreactor (AnMBR) and membrane enhanced biological phosphorous removal (MEBPR). While we observed physical EDC removal in both systems, some TH disruption activity was retained in the effluents. This work lays an important foundation for linking TH-dependent gene expression with olfactory system function in amphibians.

Modeling the exposure of wild fish to endocrine active chemicals: Potential linkages of total estrogenicity to field-observed intersex.

Decades of studies on endocrine disruption have suggested the need to manage the release of key estrogens from municipal wastewater treatment plants (WWTP). However, the proposed thresholds are below the detection limits of most routine chemical analysis, thereby restricting the ability of watershed managers to assess the environmental exposure appropriately. In this study, we demonstrated the utility of a mechanistic model to address the data gaps on estrogen exposure. Concentrations of the prominent estrogenic contaminants in wastewaters (estrone, estradiol, and ethinylestradiol) were simulated in the Grand River in southern Ontario (Canada) for nine years, including a period when major WWTP upgrades occurred. The predicted concentrations expressed as total estrogenicity (E2 equivalent concentrations) were contrasted to a key estrogenic response (i.e., intersex) in rainbow darter (Etheostoma caeruleum), a wild sentinel fish species. A predicted total estrogenicity in the river of ≥10 ng/L E2 equivalents was associated with high intersex incidence and severity, whereas concentrations <0.1 ng/L E2 equivalents were associated with minimal intersex expression. Exposure to a predicted river concentration of 0.4 ng/L E2 equivalents, the environmental quality standard (EQS) proposed by the European Union for estradiol, was associated with 34% (95% CI:30-38) intersex incidence and a very low severity score of 0.6 (95% CI:0.5-0.7). This exposure is not predicted to cause adverse effects in rainbow darter. The analyses completed in this study were only based on the predicted presence of three major estrogens (E1, E2, EE2), so caution must be exercised when interpreting the results. Nevertheless, this study illustrates the use of models for exposure assessment, especially when measured data are not available.

Multi-year prediction of estrogenicity in municipal wastewater effluents.

In this study, the estrogenicity of two major wastewater treatment plant (WWTP) effluents located in the central reaches of the Grand River watershed in southern Ontario was estimated using population demographics, excretion rates, and treatment plant-specific removals. Due to the lack of data on estrogen concentrations from direct measurements at WWTPs, the treatment efficiencies through the plants were estimated using the information obtained from an effects-directed analysis. The results show that this approach could effectively estimate the estrogenicity of WWTP effluents, both before and after major infrastructure upgrades were made at the Kitchener WWTP. The model was then applied to several possible future scenarios including population growth and river low flow conditions. The scenario analyses showed that post-upgrade operation of the Kitchener WWTP will not release highly estrogenic effluent under the 2041 projected population increase (36%) or summer low flows. Similarly, the Waterloo WWTP treatment operation is also expected to improve once the upgrades have been fully implemented and is expected to effectively treat estrogens even under extreme scenarios of population growth and river flows. The developed model may be employed to support decision making on wastewater management strategies designed for environmental protection, especially on reducing the endocrine effects in fish exposed to WWTP effluents.

Characterization of Hydrolysis Kinetics in Staged Anaerobic Digestion of Wastewater Treatment Sludge.

The hydrolysis of mixed primary and secondary sludges in two-stage anaerobic digestion was evaluated and compared with conventional single-stage digestion, using various temperature-phased configurations of M1-M2, M1-T3, T1-T2, and T1-M3. A dual hydrolysis model best described the hydrolysis in all tests. This model was also able to consistently estimate the readily and slowly fractions of particulate chemical oxygen demand (COD) of raw sludge used in the tests. The hydrolysis kinetic coefficients (Khyd_s and Khyd_r) estimated for the mesophilic digesters were significantly greater in the short hydraulic retention time (HRT) M1 digester than those of the extended HRT digesters. Conversely, at thermophilic temperatures only Khyd_r was greater in short HRT T1 digester when compared to the extended HRT digesters. The increased Khyd_r and reduced Khyd_s values due to staging effect were explained with surface reaction models and endogenous decay. The temperature dependency of Khyd_s and Khyd_r was also explored in the staged digesters.

Behavioral and molecular analyses of olfaction-mediated avoidance responses of Rana (Lithobates) catesbeiana tadpoles: Sensitivity to thyroid hormones, estrogen, and treated municipal wastewater effluent.

Olfaction is critical for survival, facilitating predator avoidance and food location. The nature of the olfactory system changes during amphibian metamorphosis as the aquatic herbivorous tadpole transitions to a terrestrial, carnivorous frog. Metamorphosis is principally dependent on the action of thyroid hormones (THs), l-thyroxine (T4) and 3,5,3'-triiodothyronine (T3), yet little is known about their influence on olfaction during this phase of postembryonic development. We exposed Taylor Kollros stage I-XIII Rana (Lithobates) catesbeiana tadpoles to physiological concentrations of T4, T3, or 17-beta-estradiol (E2) for 48h and evaluated a predator cue avoidance response. The avoidance response in T3-exposed tadpoles was abolished while T4- or E2-exposed tadpoles were unaffected compared to control tadpoles. qPCR analyses on classic TH-response gene transcripts (thra, thrb, and thibz) in the olfactory epithelium demonstrated that, while both THs produced molecular responses, T3 elicited greater responses than T4. Municipal wastewater feed stock was spiked with a defined pharmaceutical and personal care product (PPCP) cocktail and treated with an anaerobic membrane bioreactor (AnMBR). Despite substantially reduced PPCP levels, exposure to this effluent abolished avoidance behavior relative to AnMBR effluent whose feed stock was spiked with vehicle. Thibz transcript levels increased upon exposure to either effluent indicating TH mimic activity. The present work is the first to demonstrate differential TH responsiveness of the frog tadpole olfactory system with both behavioral and molecular alterations. A systems-based analysis is warranted to further elucidate the mechanism of action on the olfactory epithelium and identify further molecular bioindicators linked to behavioral response disruption.

Modeling the biotransformation of trimethoprim in biological nutrient removal system.

A pilot scale biological nutrient removal (BNR) process, batch experiments and modeling exercises were employed to investigate the removal and biotransformation of trimethoprim (TMP) in a BNR activated sludge process. The concentrations of the active microbial groups - ammonia oxidizing bacteria (AOB), ordinary heterotrophic organisms (OHOs) and polyphosphate accumulating organisms (PAOs) - in the BNR bioreactor were quantified through modeling of the pilot bioreactor. The overall TMP removal efficiency for the pilot BNR process was 64 ± 14% while the TMP biotransformation efficiencies in the anaerobic, anoxic and aerobic zones were 22 ± 20%, 27 ± 8% and 36 ± 5% respectively. Batch tests with and without nitrification inhibition showed that AOB played a role in the biotransformation of TMP in BNR activated sludge. A pseudo first order model which incorporated the contributions of PAOs, OHOs and AOB to the overall biodegradation of TMP was found to describe the biodegradation of TMP in batch tests with and without nitrification inhibition. This model showed that PAOs, OHOs and AOB contributed towards the biotransformation of TMP in aerobic BNR activated sludge with the biotransformation rate constants following the trend of kAOB > kOHOs > kPAOs.

The Effect of Solids Residence Time on Chemical Phosphorus Removal in Low Concentration Applications.

The effect of solids residence time (SRT) on steady state phosphorus (P) removal when striving for ultralow concentrations through metal salt addition was studied. Lab-scale continuous flow sequencing batch reactors (SBRs) were operated under high (6.4 mg P/L; 1.4 mol Fe/mol P) and low (3.4 mg P/L; 2.6 mol Fe/mol P) influent phosphate concentrations to characterize P removal. Residual P concentrations, particle size distribution, and microscopy analyses were determined over a range of SRTs. A majority of P removal (94% with 3.4 mg P/L; 83% with 6.4 mg P/L) occurred immediately after iron (Fe) addition with additional removal in the SBRs (3.3-4.8% with 3.4 mg P/L; 5.5-8.8% with 6.4 mg P/L). Soluble P uptake was higher for SRTs ≤ 7.4 days with 3.4 mg P/L and ≤ 14.3 days with 6.4 mg P/L. Normalized P uptake (µg P/mg total suspended solids [TSS]) decreased with SRT providing evidence that aging changed floc properties relevant to P removal. Floc size was found to have no distinguishable influence on P removal. However, changes in floc morphology were consistent with P removal trends.