Analysis of Wastewater Samples to Explore Community Substance Use in the United States: Pilot Correlative and Machine Learning Study.

BACKGROUND: Substance use disorder and associated deaths have increased in the United States, but methods for detecting and monitoring substance use using rapid and unbiased techniques are lacking. Wastewater-based surveillance is a cost-effective method for monitoring community drug use. However, the examination of the results often focuses on descriptive analysis. OBJECTIVE: The objective of this study was to explore community substance use in the United States by analyzing wastewater samples. Geographic differences and commonalities of substance use were explored. METHODS: Wastewater was sampled across the United States (n=12). Selected drugs with misuse potential, prescriptions, and over-the-counter drugs and their metabolites were tested across geographic locations for 7 days. Methods used included wastewater assessment of substances and metabolites paired with machine learning, specifically discriminant analysis and cluster analysis, to explore similarities and differences in wastewater measures. RESULTS: Geographic variations in the wastewater drug or metabolite levels were found. Results revealed a higher use of methamphetamine (z=-2.27, P=.02) and opioids-to-methadone ratios (oxycodone-to-methadone: z=-1.95, P=.05; hydrocodone-to-methadone: z=-1.95, P=.05) in states west of the Mississippi River compared to the east. Discriminant analysis suggested temazepam and methadone were significant predictors of geographical locations. Precision, sensitivity, specificity, and F1-scores were 0.88, 1, 0.80, and 0.93, respectively. Finally, cluster analysis revealed similarities in substance use among communities. CONCLUSIONS: These findings suggest that wastewater-based surveillance has the potential to become an effective form of surveillance for substance use. Further, advanced analytical techniques may help uncover geographical patterns and detect communities with similar needs for resources to address substance use disorders. Using automated analytics, these advanced surveillance techniques may help communities develop timely, tailored treatment and prevention efforts.

Surveillance of plasticizers, bisphenol A, steroids and caffeine in surface water of River Ganga and Sundarban wetland along the Bay of Bengal: occurrence, sources, estrogenicity screening and ecotoxicological risk assessment.

The transboundary River Ganga serves as a conduit for meltwater from the Himalayas and is a major freshwater source for two thirds of Indian population before emptying into the Sundarban Delta, the largest estuary in the Bay of Bengal. Endocrine disrupting compounds (EDCs) such as phthalic acid esters (PAEs) and bisphenol A (BPA) used as organic plastic additives can pollute the aquatic environment receiving plastic litter. Hence, we have investigated these EDCs in water samples from Ganga and Sundarban wetland of India. Since these compounds exhibit estrogenic potential, we have further measured steroids and evaluated the estrogenic activity (estradiol equivalents, BioE2Eqs) using an in-vitro bioassay (E-Screen). Further BioE2Eqs were compared with the sum of predicted estradiol equivalents based on the chemical concentrations of PAEs and BPA by E-Screen (ChemE2Eq) and YES factors (ChemYES). Caffeine was measured as a marker for anthropogenic wastewater discharge. Results showed that the highest BioE2Eq (below the lowest observable effect of E2 on fish) was associated with sites having sewer outfalls in the middle stretch of the river, and concomitantly coinciding with the elevated concentrations of caffeine. Neither ChemE2Eq nor ChemYES correlated with measured BioE2Eqs. River concentrations of BPA (0.04-4.46 µg/L) and ∑7plasticizers (0.43-7.63 µg/L) were higher than BPA (0.21-2.82 µg/L) and ∑7plasticizers (0.85-2 µg/L) in the Sundarban wetland. The only steroids detected were androgens, found at four sites in Ganga (0.007 µg/L± 0.003, mean ± S.D.). The highest estimated ecotoxicological risk to aquatic insect and fish stemmed from BPA. A secondary effect, and a potential impact on human health could be reflected via fish consumption from the productive fisheries region along the lower stretch of River Ganga. Identification of areas of elevated estrogenicity, plasticizer and steroid concentrations in River Ganga can be used to design and implement interventions for the remediation of such emerging contaminants.

Occurrence and removal of pharmaceutical compounds and steroids at four wastewater treatment plants in Hawai'i and their environmental fate.

The occurrence of pharmaceutical and steroid compounds in groundwater due to wastewater reuse has been reported and is of concern in tropical islands which primarily rely on groundwater. The objective of this study was to investigate the occurrence and removal of 43 pharmaceutical and steroid compounds detected in wastewater at four different wastewater treatment plants (WWTPs) in Hawai'i and to understand their environmental behavior through tropical soils as the treated effluents are used in landscapes for irrigation. Eight soil sampling locations, collected at three different depths, representing the most common soil types in Hawai'i and four WWTPs located across the major Hawaiian Islands were used. Disturbed soil samples were used to conduct the soil sorption and degradation studies and to estimate the leaching risk associated to the identified compounds. Quantification of selected compounds was conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Among the investigated compounds, only ten were detected in the treated effluents at concentrations ranging from 0.004 to 0.900 µg L-1. Caffeine (64 µg L-1) and ibuprofen (96.5 µg L-1) showed the highest concentration in raw samples, while diphenhydramine (0.9 µg L-1) showed the highest concentration in treated effluent samples. Sulfamethoxazole showed the lowest removal (0-75%). Several pharmaceuticals showed consistently higher sorption capacity and longer persistency compared with steroids regardless of soil types and depths. Poamoho (Oxisol soil) and Waimanalo (Mollisol soil) showed the highest sorption capacity, while Waimea (Entisol soil) showed the lowest sorption capacity. Soil physico-chemical properties (i.e., clay content, level of organic carbon, and presence of metal oxide) and soil depth highly impacted the sorption behavior of the selected pharmaceutical compounds. In particular, the sorption capacity decreased with soil depth due to the higher level of organic carbon present in the first 30 cm compared with the deeper depths (60-90 cm).

Estrogenic effects following larval exposure to the putative anti-estrogen, fulvestrant, in the fathead minnow (Pimephales promelas).

The objective of the present study was to investigate the consequences of early-life exposure to fulvestrant on estrogenic gene expression in fathead minnow larvae. To address this objective, fathead minnow larvae were exposed to fulvestrant (ICI 182,780) during the window of sexual differentiation between 0 to 30 days post-hatch (dph). The four treatment groups in this study included: filtered water controls (never exposed), solvent controls (ethanol 0.01%), and nominally low (0.10µg/L) and high (10.0µg/L) doses of fulvestrant. Following 30 d exposure to their respective treatment, larvae were transferred to filtered water aquaria and assessed for alterations in endocrine-responsive gene expression (i.e., RT-qPCR), body size and survival. The remaining fish depurated in filtered water until reaching sexual maturity (180dph) for assessment of persistent effects on sex characteristics, reproductive performance and sex ratio. Following the 30-d early life exposure, larvae showed upregulations of the endocrine-responsive genes ar, erß and vtg in response to both low and high doses of fulvestrant, but showed no differences in survival or body mass. Upon reaching sexual maturity under depuration conditions, male minnows previously exposed to fulvestrant as larvae showed reductions in gonad mass along with the feminization of secondary sex characteristics with no observed effects in females. Exposure to fulvestrant had no effects on gonadal histology, reproductive performance or final sex ratio as adults. Results from this study demonstrate that aqueous exposure to fulvestrant is estrogenic in fathead minnow larvae and is capable of feminizing male fish as adults following early life exposure.

Sensitive and simplified analysis of natural and synthetic steroids in water and solids using on-line solid-phase extraction and microwave-assisted solvent extraction coupled to liquid chromatography tandem mass spectrometry atmospheric pressure photoionization.

Analytical methods for the determination of several natural and synthetic steroids in water and solid samples were developed using liquid chromatography tandem mass spectrometry. Atmospheric pressure photoionization (APPI) in positive mode was used as the source which appeared to reduce background and interferences as compared to electrospray ionization. Toluene was identified as the most appropriate dopant and its concentration optimized to enhance ionization efficiency. Method detection limits were in the low-nanogram-per-liter range for water samples using on-line solid-phase extraction and near 0.2 ng/g using microwave-assisted solvent extraction for solid samples. This on-line method, using APPI as a source and toluene as a dopant, is beneficial as it saves time due to on-line extraction and reduces interference and matrix effects using gas-phase photoionization. Groundwater, wastewater, runoff, and soil samples were collected and analyzed using the method indicating that traces of several androgens, estrogens, and other steroid-like compounds do occur in samples impacted by animal waste.

Using gas-phase molecular descriptors to predict dechlorination rates of chloroalkanes by zerovalent iron.

Reductive dehalogenation of chlorinated compounds is the most important process occurring within the zerovalent iron (Fe0) barrier. The relative reaction rates of individual halocarbons with Fe0 can vary considerably. This variability has been the stimulus for using various chemical descriptors for a priori predictions of transformation rates via linear free-energy relationships (LFERs). Our objective was to determine the efficacy of four molecular descriptors to describe the transformation rates of three chloromethanes, three chloroethanes, and six chloropropanes by Fe0. This was accomplished by generating an internally consistent set of rate constants under controlled environmental conditions (16 degrees C, anaerobic) and regressing the surface-area normalized rate constants (k(SA)) against (i) energy of the lowest unoccupied molecular orbital (E(LUMO)); (ii) vertical attachment energies (VAE); (iii) thermal electron attachment rate constants; and (iv) the molar response from a commercial electron capture detector (ECD). Results showed good correlations between k(SA)'s and all four descriptors (r2: 0.72-1.0), but a separate trend line was required for the chloromethanes and the chloro- ethanes/ propanes. Given the availability and ease with which ECD response can be obtained, this physical measurement may provide a practical means of determining relative rates of reactivity of various halocarbons in permeable reactive iron barriers.

Electron capture detector response and dissociative electron attachment cross sections in chloroalkanes and chloroalkenes.

Electron capture detectors (ECDs) are widely used in gas chromatography to detect electronegative compounds. In this work, we examine the connections between the ECD response and the cross sections for dissociative electron attachment (DEA) determined from low energy electron beam studies in the chloroalkane family, stressing in particular the role of temporary anion state energies. We show that attachment rate coefficients computed from these cross sections are well correlated with ECD response, and that the latter decreases exponentially with increasing energies of the lowest anion states. ECD measurements are also carried out in monochloroalkanes substituted with unsaturated ethenyl and phenyl moieties, and the response is shown to depend strongly on the mixing between the unsaturated pi* and the C-Cl sigma* temporary anions as exhibited by the vertical attachment energies (VAEs) of these states. The results show good correlations between the chloroalkene and phenyl chloride ECD responses and the VAEs for the mixed states.