Black Carbon Impacts on Paraburkholderia xenovorans Strain LB400 Cell Enrichment and Activity: Implications toward Lower-Chlorinated Polychlorinated Biphenyls Biodegradation Potential.

Volatilization of lower-chlorinated polychlorinated biphenyls (LC-PCBs) from sediment poses health threats to nearby communities and ecosystems. Biodegradation combined with black carbon (BC) materials is an emerging bioaugmentation approach to remove PCBs from sediment, but development of aerobic biofilms on BC for long-term, sustained LC-PCBs remediation is poorly understood. This work aimed to characterize the cell enrichment and activity of biphenyl- and benzoate-grown Paraburkholderia xenovorans strain LB400 on various BCs. Biphenyl dioxygenase gene (bphA) abundance on four BC types demonstrated corn kernel biochar hosted at least 4 orders of magnitude more attached cells per gram than other feedstocks, and microscopic imaging revealed the attached live cell fraction was >1.5× more on corn kernel biochar than GAC. BC characteristics (i.e., sorption potential, pore size, pH) appear to contribute to cell attachment differences. Reverse transcription qPCR indicated that BC feedstocks significantly influenced bphA expression in attached cells. The bphA transcript-per-gene ratio of attached cells was >10-fold more than suspended cells, confirmed by transcriptomics. RNA-seq also demonstrated significant upregulation of biphenyl and benzoate degradation pathways on attached cells, as well as revealing biofilm formation potential/cell-cell communication pathways. These novel findings demonstrate aerobic PCB-degrading cell abundance and activity could be tuned by adjusting BC feedstocks/attributes to improve LC-PCBs biodegradation potential.

Functional Group Properties and Position Drive Differences in Xenobiotic Plant Uptake Rates, but Metabolism Shares a Similar Pathway.

Plant uptake of xenobiotic compounds is crucial for phytoremediation (including green stormwater infrastructure) and exposure potential during crop irrigation with recycled water. Experimentally determining the plant uptake for every relevant chemical is impractical; therefore, illuminating the role of specific functional groups on the uptake of trace organic contaminants is needed to enhance predictive power. We used benzimidazole derivatives to probe the impact of functional group electrostatic properties and position on plant uptake and metabolism using the hydroponic model plant Arabidopsis thaliana. The greatest plant uptake rates occurred with an electron-withdrawing functional group at the 2 position; however, uptake was still observed with an electron-donating group. An electron-donating group at the 1 position significantly slowed uptake for both benzimidazole- and benzotriazole-based molecules used in this study, indicating possible steric effects. For unsubstituted benzimidazole and benzotriazole structures, the additional heterocyclic nitrogen in benzotriazole increased plant uptake rates compared to benzimidazole. Analysis of quantitative structure-activity relationship parameters for the studied compounds implicates energy-related molecular descriptors as uptake drivers. Despite significantly varied uptake rates, compounds with different functional groups yielded shared metabolites, including an impact on endogenous glutathione production. Although the topic is complex and influenced by multiple factors in the field, this study provides insights into the impact of functional groups on plant uptake, with implications for environmental fate and consumer exposure.

Microbial Biotransformation Products and Pathways of Dichloroacetamide Herbicide Safeners.

Dichloroacetamide safeners are common ingredients in commercial herbicide formulations. We previously investigated the environmental fate of dichloroacetamides via photolysis and hydrolysis, but other potentially important, environmentally relevant fate processes remain uncharacterized and may yield products of concern. Here, we examined microbial biotransformation of two dichloroacetamide safeners, benoxacor and dichlormid, to identify products and elucidate pathways. Using aerobic microcosms inoculated with river sediment, we demonstrated that microbial biotransformations of benoxacor and dichlormid proceed primarily, if not exclusively, via cometabolism. Benoxacor was transformed by both hydrolysis and microbial biotransformation processes; in most cases, biotransformation rates were faster than hydrolysis rates. We identified multiple novel products of benoxacor and dichlormid not previously observed for microbial processes, with several products similar to those reported for structurally related chloroacetamide herbicides, thus indicating potential for conserved biotransformation mechanisms across both chemical classes. Observed products include monochlorinated species such as the banned herbicide CDAA (from dichlormid), glutathione conjugates, and sulfur-containing species. We propose a transformation pathway wherein benoxacor and dichlormid are first dechlorinated, likely via microbial hydrolysis, and subsequently conjugated with glutathione. This is the first study reporting biological dechlorination of dichloroacetamides to yield monochlorinated products in aerobic environments.

Transcriptome signatures of wastewater effluent exposure in larval zebrafish vary with seasonal mixture composition in an effluent-dominated stream.

Wastewater treatment plant (WWTP) effluent-dominated streams provide critical habitat for aquatic and terrestrial organisms but also continually expose them to complex mixtures of pharmaceuticals that can potentially impair growth, behavior, and reproduction. Currently, few biomarkers are available that relate to pharmaceutical-specific mechanisms of action. In the experiment reported in this paper, zebrafish (Danio rerio) embryos at two developmental stages were exposed to water samples from three sampling sites (0.1 km upstream of the outfall, at the effluent outfall, and 0.1 km below the outfall) during base-flow conditions from two months (January and May) of a temperate-region effluent-dominated stream containing a complex mixture of pharmaceuticals and other contaminants of emerging concern. RNA-sequencing identified potential biological impacts and biomarkers of WWTP effluent exposure that extend past traditional markers of endocrine disruption. Transcriptomics revealed changes to a wide range of biological functions and pathways including cardiac, neurological, visual, metabolic, and signaling pathways. These transcriptomic changes varied by developmental stage and displayed sensitivity to variable chemical composition and concentration of effluent, thus indicating a need for stage-specific biomarkers. Some transcripts are known to be associated with genes related to pharmaceuticals that were present in the collected samples. Although traditional biomarkers of endocrine disruption were not enriched in either month, a high estrogenicity signal was detected upstream in May and implicates the presence of unidentified chemical inputs not captured by the targeted chemical analysis. This work reveals associations between bioeffects of exposure, stage of development, and the composition of chemical mixtures in effluent-dominated surface water. The work underscores the importance of measuring effects beyond the endocrine system when assessing the impact of bioactive chemicals in WWTP effluent and identifies a need for non-targeted chemical analysis when bioeffects are not explained by the targeted analysis.

Sensitive and Specific Detection of Estrogens Featuring Doped Silicon Nanowire Arrays.

Estrogens and estrogen-mimicking compounds in the aquatic environment are known to cause negative impacts to both ecosystems and human health. In this initial proof-of-principle study, we developed a novel vertically oriented silicon nanowire (vSiNW) array-based biosensor for low-cost, highly sensitive and selective detection of estrogens. The vSiNW arrays were formed using an inexpensive and scalable metal-assisted chemical etching (MACE) process. A vSiNW array-based p-n junction diode (vSiNW-diode) transducer design for the biosensor was used and functionalized via 3-aminopropyltriethoxysilane (APTES)-based silane chemistry to bond estrogen receptor-alpha (ER-α) to the surface of the vSiNWs. Following receptor conjugation, the biosensors were exposed to increasing concentrations of estradiol (E2), resulting in a well-calibrated sensor response (R 2 ≥ 0.84, 1-100 ng/mL concentration range). Fluorescence measurements quantified the distribution of estrogen receptors across the vSiNW array compared to planar Si, indicating an average of 7 times higher receptor presence on the vSiNW array surface. We tested the biosensor's target selectivity by comparing it to another estrogen (estrone [E1]) and an androgen (testosterone), where we measured a high positive electrical biosensor response after E1 exposure and a minimal response after testosterone. The regeneration capacity of the biosensor was tested following three successive rinses with phosphate buffer solution (PBS) between hormone exposure. Traditional horizontally oriented Si NW field effect transistor (hSiNW-FET)-based biosensors report electrical current changes at the nanoampere (nA) level that require bulky and expensive measurement equipment making them unsuitable for field measurements, whereas the reported vSiNW-diode biosensor exhibits current changes in the microampere (µA) range, demonstrating up to 100-fold electrical signal amplification, thus enabling sensor signal measurement using inexpensive electronics. The highly sensitive and specific vSiNW-diode biosensor developed here will enable the creation of low-cost, portable, field-deployable biosensors that can detect estrogenic compounds in waterways in real-time.

Use of pilot-scale geomedia-amended biofiltration system for removal of polar trace organic and inorganic contaminants from stormwater runoff.

Stormwater runoff capture and groundwater recharge can provide a sustainable means of augmenting the local water resources in water-stressed cities while simultaneously mitigating flood risk, provided that these processes do not compromise groundwater quality. We developed and tested for one year an innovative pilot-scale stormwater treatment train that employs cost-effective engineered geomedia in a continuous-flow unit-process system to remove contaminants from urban runoff during aquifer recharge. The system consisted of an iron-enhanced sand filter for phosphate removal, a woodchip bioreactor for nitrate removal coupled to an aeration step, and columns packed with different configurations of biochar- and manganese oxide-containing sand to remove trace metals and persistent, mobile, and toxic trace organic contaminants. During conditioning with authentic stormwater runoff over an extended period (8 months), the woodchip bioreactor removed 98% of the influent nitrate (9 g-N m-3 d-1), while phosphate broke through the iron-enhanced sand filter. During the challenge test (4 months), geomedia removed more than 80% of the mass of metals and trace organic compounds. Column hydraulic performance was stable during the entire study, and the weathered biochar and manganese oxide were effective at removing trace organic contaminants and metals, respectively. Under conditions likely encountered in the field, sustained nutrient removal is probable, but polar organic compounds such as 2,4-D could breakthrough after about a decade for conditions at the study site.

Modeling Risk Dynamics of Contaminants of Emerging Concern in a Temperate-region Wastewater Effluent-dominated Stream.

Wastewater effluent-dominated streams are becoming increasingly common worldwide, including in temperate regions, with potential impacts on ecological systems and drinking water sources. We recently quantified the occurrence/ spatiotemporal dynamics of pharmaceutical mixtures in a representative temperate-region wastewater effluent-dominated stream (Muddy Creek, Iowa) under baseflow conditions and characterized relevant fate processes. Herein, we quantified the ecological risk quotients (RQs) of 19 effluent-derived contaminants of emerging concern (CECs; including: 14 pharmaceuticals, 2 industrial chemicals, and 3 neonicotinoid insecticides) and 1 run-off-derived compound (atrazine) in the stream under baseflow conditions, and estimated the probabilistic risks of effluent-derived CECs under all-flow conditions (i.e., including runoff events) using stochastic risk modeling. We determined that 11 out of 20 CECs pose medium-to-high risks to local ecological systems (i.e., algae, invertebrates, fish) based on literature-derived acute effects under measured baseflow conditions. Stochastic risk modeling indicated decreased, but still problematic, risk of effluent-derived CECs (i.e., RQ≥0.1) under all-flow conditions when runoff events were included. Dilution of effluent-derived chemicals from storm flows thus only minimally decreased risk to aquatic biota in the effluent-dominated stream. We also modeled in-stream transport. Thirteen out of 14 pharmaceuticals persisted along the stream reach (median attenuation rate constant k<0.1 h-1) and entered the Iowa River at elevated concentrations. Predicted and measured concentrations in the drinking water treatment plant were below the human health benchmarks. This study demonstrates the application of probabilistic risk assessments for effluent-derived CECs in a representative effluent-dominated stream under variable flow conditions (when measurements are less practical) and provides an enhanced prediction tool transferable to other effluent-dominated systems.

RNA-seq reveals potential gene biomarkers in fathead minnows (Pimephales promelas) for exposure to treated wastewater effluent.

Discharged wastewater treatment plant (WWTP) effluent greatly contributes to the generation of complex mixtures of contaminants of emerging concern (CECs) in aquatic environments which often contain neuropharmaceuticals and other emerging contaminants that may impact neurological function. However, there is a paucity of knowledge on the neurological impacts of these exposures to aquatic organisms. In this study, caged fathead minnows (Pimephales promelas) were exposed in situ in a temperate-region effluent-dominated stream (i.e., Muddy Creek) in Coralville, Iowa, USA upstream and downstream of a WWTP effluent outfall. The pharmaceutical composition of Muddy Creek was recently characterized by our team and revealed many compounds there were at a low microgram to high nanogram per liter concentration. Total RNA sequencing analysis on brain tissues revealed 280 gene isoforms that were significantly differentially expressed in male fish and 293 gene isoforms in female fish between the upstream and downstream site. Only 66 (13%) of such gene isoforms overlapped amongst male and female fish, demonstrating sex-dependent impacts on neuronal gene expression. By using a systems biology approach paired with functional enrichment analyses, we identified several potential novel gene biomarkers for treated effluent exposure that could be used to expand monitoring of environmental effects with respect to complex CEC mixtures. Lastly, when comparing the results of this study to those that relied on a single-compound approach, there was relatively little overlap in terms of gene-specific effects. This discovery brings into question the application of single-compound exposures in accurately characterizing environmental risks of complex mixtures and for gene biomarker identification.

Rapid plant uptake of isothiazolinone biocides and formation of metabolites by hydroponic Arabidopsis.

Isothiazolinones biocides are water-soluble, low molecular weight, nitrogenous compounds widely used to prevent microbial growth in a variety of applications including personal care products and building façade materials. Because isothiazolinones from buildings wash off and enter stormwater, interactions with terrestrial plants may represent an important part of the environmental fate of these compounds (e.g., in green stormwater infrastructure). Using the model plant Arabidopsis thaliana grown hydroponically, we observed rapid (≥99% within 24 hours), plant-driven removal of four commonly used isothiazolinones: benzisothiazolinone (BIT), chloromethylisothiazolinone, methylisothiazolinone, and octylisothiazolinone. No significant differences in uptake rate occurred between the four compounds; therefore, BIT was used for further detailed investigation. BIT uptake by Arabidopsis was concentration-dependent in a manner that implicates transporter-mediated substrate inhibition. BIT uptake was also minimally impacted by multiple BIT spikes, suggesting constituently active uptake. BIT plant uptake rate was robust, unaffected by multiple inhibitors. We investigated plant metabolism as a relevant removal process. Proposed major metabolites that significantly increased in the BIT-exposure treatment compared to the control included: endogenous plant compounds nicotinic acid (confirmed with a reference standard) and phenylthioacetohydroximic acid, a possible amino acid-BIT conjugate, and two accurate masses of interest. Two of the compounds (phenylthioacetohydroximic acid and TP 470) were also present in increased amounts in the hydroponic medium after BIT exposure, possibly via plant excretion. Upregulation of endogenous plant compounds is environmentally significant because it demonstrates that BIT impacts plant biology. The rapid plant-driven isothiazolinone removal observed here indicates that plant-isothiazolinone processes could be relevant to the environmental fate of these stormwater compounds.

Harnessing plant-microbiome interactions for bioremediation across a freshwater urbanization gradient.

Urbanization impacts land, air, and water, creating environmental gradients between cities and rural areas. Urban stormwater delivers myriad co-occurring, understudied, and mostly unregulated contaminants to aquatic ecosystems, causing a pollution gradient. Recipient ecosystems host interacting species that can affect each others' growth and responses to these contaminants. For example, plants and their microbiomes often reciprocally increase growth and contaminant tolerance. Here, we identified ecological variables affecting contaminant fate across an urban-rural gradient using 50 sources of the aquatic plant Lemna minor (duckweed) and associated microbes, and two co-occurring winter contaminants of temperate cities, benzotriazole and salt. We conducted experiments totalling >2,500 independent host-microbe-contaminant microcosms. Benzotriazole and salt negatively affected duckweed growth, but not microbial growth, and duckweeds maintained faster growth with their local, rather than disrupted, microbiota. Benzotriazole transformation products of plant, microbial, and phototransformation pathways were linked to duckweed and microbial growth, and were affected by salt co-contamination, microbiome disruption, and source sites of duckweeds and microbes. Duckweeds from urban sites grew faster and enhanced phytotransformation, but supported less total transformation of benzotriazole. Increasing microbial community diversity correlated with greater removal of benzotriazole, but taxonomic groups may explain shifts across transformation pathways: the genus Aeromonas was linked to increasing phototransformation. Because benzotriazole toxicity could depend on amount and type of in situ transformation, this variation across duckweeds and microbes could be harnessed for better management of urban stormwater. Broadly, our results demonstrate that plant-microbiome interactions harbour manipulable variation for bioremediation applications.

White Rot Fungi Produce Novel Tire Wear Compound Metabolites and Reveal Underappreciated Amino Acid Conjugation Pathways.

There is increasing concern about tire wear compounds (TWCs) in surface water and stormwater as evidence grows on their toxicity and widespread detection in the environment. Because TWCs are prevalent in stormwater, there is a need to understand fate and treatment options including biotransformation in green infrastructure (e.g., bioretention). Particularly, fungal biotransformation is not well-studied in a stormwater context despite the known ability of certain fungi to remove recalcitrant contaminants. Here, we report the first study on fungal biotransformation of the TWCs acetanilide and hexamethoxymethylmelamine (HMMM). We found that the model white rot fungus, Trametes versicolor, removed 81.9% and 69.6% of acetanilide and HMMM, respectively, with no significant sorption to biomass. The bicyclic amine 1,3-diphenylguanidine was not removed. Additionally, we identified novel TWC metabolites using semi-untargeted metabolomics via high-resolution mass spectrometry. Key metabolites include multiple isomers of HMMM biotransformation products, melamine as a possible "dead-end" product of HMMM (verified with an authentic standard), and a glutamine-conjugated product of acetanilide. These metabolites have implications for environmental toxicity and treatment. Our discovery of the first fungal glutamine-conjugated product highlights the need to investigate amino acid conjugation as an important pathway in biotransformation of contaminants, with implications in other fields including natural products discovery.

The use of recycled materials in a biofilter to polish anammox wastewater treatment plant effluent.

Anammox is gaining popularity for treating wastewater containing high-strength ammonia due to lower energy demand compared to conventional nitrification-denitrification processes; however, anammox is reported to increase nitrate loads in the effluent. The objective of this study was to assess the applicability of recycled materials [recycled concrete aggregate (RCA) and rice husks (RH)] as a polishing step to improve anammox reactor effluent quality. Anammox effluents were separately passed through two single-stage columns containing RCA and RH, and one two-stage column (50% RCA, 50% RH) to quantify total N, ammonia, nitrate, nitrite, and phosphate removal efficiencies. Langmuir isotherm experiments were conducted to quantify nitrate, nitrite, and phosphate sorption capacities in the columns. The RCA column exhibited the highest phosphate sorption capacity (0.074 mg/g), while the RH column exhibited higher nitrite and nitrate adsorption (0.063 mg/g and 0.023 mg/g respectively). We created a Hydrus-1D model to estimate pseudo-first-order reaction rates in the columns. Because RCA media can form metal-phosphate precipitates, the fastest phosphate reaction rate (1.58 min-1) occurred in the RCA column. The two-stage column demonstrated the greatest overall removals for all nutrients, and removal rates were consistent throughout the experimental period. The two-stage column achieved 15% total N, 94% ammonia-N, 38% nitrate-N, 75% nitrite-N, and 27% phosphate removal. The maximum nitrite, nitrate, and phosphate adsorption capacities in the two-stage column were 0.030 mg/g, 0.017 mg/g, and 0.014 mg/g respectively. This is the first study to demonstrate that recycled materials can successfully be integrated into a biofilter as an effluent polishing step to remove nutrients from anammox wastewater.

Combining Experimental Sorption Parameters with QSAR to Predict Neonicotinoid and Transformation Product Sorption to Carbon Nanotubes and Granular Activated Carbon.

We recently discovered that transformation of the neonicotinoid insecticidal pharmacophore alters sorption propensity to activated carbon, with products adsorbing less than parent compounds. To assess the environmental fate of novel transformation products that lack commercially available standards, researchers must rely on predictive approaches. In this study, we combined computationally derived quantitative structure-activity relationship (QSAR) parameters for neonicotinoids and neonicotinoid transformation products with experimentally determined Freundlich partition constants (log K F for sorption to carbon nanotubes [CNTs] and granular activated carbon [GAC]) to model neonicotinoid and transformation product sorption. QSAR models based on neonicotinoid sorption to functionalized/nonfunctionalized CNTs (used to generalize/simplify neonicotinoid-GAC interactions) were iteratively generated to obtain a multiple linear regression that could accurately predict neonicotinoid sorption to CNTs using internal and external validation (within 0.5 log units of the experimentally determined value). The log K F,CNT values were subsequently related to log K F,GAC where neonicotinoid sorption to GAC was predicted within 0.3 log-units of experimentally determined values. We applied our neonicotinoid-specific model to predict log K F,GAC for a suite of novel neonicotinoid transformation products (i.e., formed via hydrolysis, biotransformation, and chlorination) that do not have commercially available standards. We present this modeling approach as an innovative yet relatively simple technique to predict fate of highly specialized/unique polar emerging contaminants and/or transformation products that cannot be accurately predicted via traditional methods (e.g., pp-LFER), and highlights molecular properties that drive interactions of emerging contaminants.

Acid- and Base-Mediated Hydrolysis of Dichloroacetamide Herbicide Safeners.

Safeners are used extensively in commercial herbicide formulations. Although safeners are regulated as inert ingredients, some of their transformation products have enhanced biological activity. Here, to fill gaps in our understanding of safener environmental fate, we determined rate constants and transformation products associated with the acid- and base-mediated hydrolysis of dichloroacetamide safeners AD-67, benoxacor, dichlormid, and furilazole. Second-order rate constants for acid- (HCl) and base-mediated (NaOH) dichloroacetamide hydrolysis (2.8 × 10-3 to 0.46 and 0.3-500 M-1 h-1, respectively) were, in many cases (5 of 8), greater than those reported for their chloroacetamide herbicide co-formulants. In particular, the rate constant for base-mediated hydrolysis of benoxacor was 2 orders of magnitude greater than that of its active ingredient co-formulant, S-metolachlor. At circumneutral pH, only benoxacor underwent appreciable hydrolysis (5.3 × 10-4 h-1), and under high-pH conditions representative of lime-soda softening, benoxacor's half-life was 13 h─a timescale consistent with partial transformation during water treatment. Based on Orbitrap LC-MS/MS analysis of dichloroacetamide hydrolysis product mixtures, we propose structures for major products and three distinct mechanistic pathways that depend on the system pH and compound structure. These include base-mediated amide cleavage, acid-mediated amide cleavage, and acid-mediated oxazolidine ring opening. Collectively, this work will help to identify systems in which hydrolysis contributes to the transformation of dichloroacetamides, while also highlighting important differences in the reactivity of dichloroacetamides and their active chloroacetamide co-formulants.

Exposure and Transport of Alkaloids and Phytoestrogens from Soybeans to Agricultural Soils and Streams in the Midwestern United States.

Phytotoxins are naturally produced toxins with potencies similar/higher than many anthropogenic micropollutants. Nevertheless, little is known regarding their environmental fate and off-field transport to streams. To fill this research gap, a network of six basins in the Midwestern United States with substantial soybean production was selected for the study. Stream water (n = 110), soybean plant tissues (n = 8), and soil samples (n = 16) were analyzed for 12 phytotoxins (5 alkaloids and 7 phytoestrogens) and 2 widely used herbicides (atrazine and metolachlor). Overall, at least 1 phytotoxin was detected in 82% of the samples, with as many as 11 phytotoxins detected in a single sample (median = 5), with a concentration range from below detection to 37 and 68 ng/L for alkaloids and phytoestrogens, respectively. In contrast, the herbicides were ubiquitously detected at substantially higher concentrations (atrazine: 99% and metolachlor: 83%; the concentrations range from below detection to 150 and 410 ng/L, respectively). There was an apparent seasonal pattern for phytotoxins, where occurrence prior to and during harvest season (September to November) and during the snow melt season (March) was higher than that in December-January. Runoff events increased phytotoxin and herbicide concentrations compared to those in base-flow conditions. Phytotoxin plant concentrations were orders of magnitude higher compared to those measured in soil and streams. These results demonstrate the potential exposure of aquatic and terrestrial organisms to soybean-derived phytotoxins.

Tandem field and laboratory approaches to quantify attenuation mechanisms of pharmaceutical and pharmaceutical transformation products in a wastewater effluent-dominated stream.

Evolving complex mixtures of pharmaceuticals and transformation products in effluent-dominated streams pose potential impacts to aquatic species; thus, understanding the attenuation dynamics in the field and characterizing the prominent attenuation mechanisms of pharmaceuticals and their transformation products (TPs) is critical for hazard assessments. Herein, we determined the attenuation dynamics and the associated prominent mechanisms of pharmaceuticals and their corresponding TPs via a combined long-term field study and controlled laboratory experiments. For the field study, we quantified spatiotemporal exposure concentrations of five pharmaceuticals and six associated TPs in a small, temperate-region effluent-dominated stream during baseflow conditions where the wastewater plant was the main source of pharmaceuticals. We selected four sites (upstream, at, and two progressively downstream from effluent discharge) and collected water samples at 16 time points (64 samples in total, approximately twice monthly, depending on flows) for 1 year. Concurrently, we conducted photolysis, sorption, and biodegradation batch tests under controlled conditions to determine the major attenuation mechanisms. We observed 10-fold greater attenuation rates in the field compared to batch tests, demonstrating that connecting laboratory batch tests with field measurements to enhance predictive power is a critical need. Batch systems alone, often used for assessment, are useful for determining fate processes but poorly approximate in-stream attenuation kinetics. Sorption was the dominant attenuation process (t1/2<7.7 d) for 5 of 11 compounds in the batch tests, while the other compounds (n = 6) persisted in the batch tests and along the 5.1 km stream reach. In-stream parent-to-product transformation was minimal. Differential attenuation contributed to the evolving pharmaceutical mixture and created changing exposure conditions with concomitant implications for aquatic and terrestrial biota. Tandem field and laboratory characterization can better inform modeling efforts for transport and risk assessments.

Emerging investigator series: municipal wastewater as a year-round point source of neonicotinoid insecticides that persist in an effluent-dominated stream.

Neonicotinoids in aquatic systems have been predominantly associated with agriculture, but some are increasingly being linked to municipal wastewater. Thus, the aim of this work was to understand the municipal wastewater contribution to neonicotinoids in a representative, characterized effluent-dominated temperate-region stream. Our approach was to quantify the spatiotemporal concentrations of imidacloprid, clothianidin, thiamethoxam, and transformation product imidacloprid urea: 0.1 km upstream, the municipal wastewater effluent, and 0.1 and 5.1 km downstream from the wastewater outfall (collected twice-monthly for one year under baseflow conditions). Quantified results demonstrated that wastewater effluent was a point-source of imidacloprid (consistently) and clothianidin (episodically), where chronic invertebrate exposure benchmarks were exceeded for imidacloprid (36/52 samples; 3/52 > acute exposure benchmark) and clothianidin (8/52 samples). Neonicotinoids persisted downstream where mass loads were not significantly different than those in the effluent. The combined analysis of neonicotinoid effluent concentrations, instream seasonality, and registered uses in Iowa all indicate imidacloprid, and seasonally clothianidin, were driven by wastewater effluent, whereas thiamethoxam and imidacloprid urea were primarily from upstream non-point sources (or potential in-stream transformation for imidacloprid urea). This is the first study to quantify neonicotinoid persistence in an effluent-dominated stream throughout the year-implicating wastewater effluent as a point-source for imidacloprid (year-round) and clothianidin (seasonal). These findings suggest possible overlooked neonicotinoid indoor human exposure routes with subsequent implications for instream ecotoxicological exposure.

Improving the spatial and temporal monitoring of cyanotoxins in Iowa lakes using a multiscale and multi-modal monitoring approach.

Cyanobacterial harmful algal blooms (CyanoHABs) are pervasive and negatively impact lake water quality, resulting in economic losses and public health risks through exposure to cyanotoxins. Therefore, it is critical to better monitor and understand the complexity of CyanoHABs, but current methods do not fully describe the spatial and temporal variability of bloom events. In this work, we developed a framework for a multiscale and multi-modal monitoring approach for CyanoHABs combining drone-based near-range remote sensing with analytical measurements of microcystin cyanotoxins and chlorophyll-a. We analyzed weekly beach monitoring samples from 37 lakes geographically distributed across the state of Iowa (USA) over a 15-week period in the summer of 2019 to quantify ELISA (bioassay), 12 microcystin congeners (LC-MS/MS), and chlorophyll-a. We developed a novel microcystin congener-normalized equivalent toxin metric to compare CyanoHAB impacted waters; this microcystin-LR normalized sum-of-congeners approach yields lower predicted toxicity than parallel ELISA results suggesting ELISA is conservative for assessment. A significant linear relationship existed between chlorophyll-a and microcystin for lakes throughout Iowa (R2 = 0.39, p < 0.001); lakes with low watershed:lake area ratio and long residence times exhibited a stronger correlation. We then developed a novel geometry-based image processing approach to allow for stitching over-water drone images, a previous barrier in photogrammetry. We applied our mutli-modal framework to a case study on Green Valley Lake to assess initial viability and predicted microcystin concentrations within 33%. We concluded that multispectral imaging is possible but may presently be insufficient for predicting microcystin concentrations due to limitations in the spectral capabilities of the multispectral camera, but technologies are quickly advancing, and lightweight hyperspectral imaging could soon become feasible for investigating spatial bloom variability on lakes.

Differences in Neonicotinoid and Metabolite Sorption to Activated Carbon Are Driven by Alterations to the Insecticidal Pharmacophore.

Widespread application of neonicotinoids has led to their proliferation in waters. Despite low neonicotinoid hydrophobicity, our prior studies implicated granular activated carbon (GAC) in neonicotinoid removal. Based on known receptor binding characteristics, we hypothesized that the insecticidal pharmacophore influences neonicotinoid sorption. Our objectives were to illuminate drivers of neonicotinoid sorption for parent neonicotinoids (imidacloprid, clothianidin, thiamethoxam, and thiacloprid) and pharmacophore-altered metabolites (desnitro-imidacloprid and imidacloprid urea) to GAC, powdered activated carbon, and carbon nanotubes (CNTs). Neonicotinoid sorption to GAC was extensive and largely irreversible, with significantly greater sorption of imidacloprid than desnitro-imidacloprid. Imidacloprid and imidacloprid urea (electronegative pharmacophores) sorbed most extensively to nonfunctionalized CNTs, whereas desnitro-imidacloprid (positive pharmacophore) sorbed most to COOH-CNTs, indicating the importance of charge interactions and/or hydrogen bonding between the pharmacophore and carbon surface. Water chemistry parameters (temperature, alkalinity, ionic strength, and humic acid) inhibited overall neonicotinoid sorption, suggesting that pharmacophore-driven sorption in real waters may be diminished. Analysis of a full-scale drinking water treatment plant GAC filter influent, effluent, and spent GAC attributes neonicotinoid/metabolite removal to GAC under real-world conditions for the first time. Our results demonstrate that the neonicotinoid pharmacophore not only confers insecticide selectivity but also impacts sorption behavior, leading to less effective removal of metabolites by GAC filters in water treatment.