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Knowledge of the number of people present in a catchment is fundamental for the assessment of spatio-temporal trends in wastewater-based epidemiology (WBE). Accurately estimating the number of people connected to wastewater catchments is challenging however, because populations are dynamic. Methods used to estimate population size can significantly influence the calculation and interpretation of population-normalised wastewater data (PNWD). This paper systematically reviews the reporting of population data in 339 WBE studies. Studies were evaluated based on their reporting of population size, the source of population data, the population calculation methods, and the uncertainties in population estimates. Most papers reported population size (96 %) and the source of population data (60 %). Fewer studies reported the uncertainties in their population data (50 %) and the methods used to calculate these estimates (28 %). This is relevant because different methods have unique strengths and limitations which can affect the accuracy of PNWD. Only 64 studies (19 %) reported all four components of population data. The reporting of population data has remained consistent in the past decade. Based on the findings, we recommend generalised reporting criteria for population data in WBE. As WBE is further mainstreamed and applied, the clear and comprehensive reporting of population data will only become increasingly important., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background and Aims: Between 2018 and 2020, Australia implemented major policy changes to improve the quality and safety of opioid prescribing, with a specific focus on oxycodone. This study used wastewater-based epidemiology to assess the efficacy of Australia's regulatory reforms by measuring change in consumption of oxycodone via exploratory analysis., Design, Setting, Participants, Measurements: Wastewater analysis data on oxycodone consumption was from the National Wastewater Drug Monitoring Program. The program captures data from more than 50 wastewater treatment plant catchments across Australia, equivalent to more than 50% of the national population. Geographic trend analyses were conducted for both major cities and regional areas within all states and territories of Australia over a 6-year period between 2017 and 2023., Findings: Oxycodone consumption showed a statistically significant increase nationally from 78 mg/day/1000 people (95% confidence interval [CI] = 71, 84) in 2017 to 120 mg/day/1000 people in August 2019 (95% CI = 110, 120), an increase of 52% (95% CI = 42, 62, P < 0.0001). From August 2019 to December 2020, there was a statistically significant decrease from 120 to 65 mg/day/1000 people (95% CI = 60, 71), a decrease of 45% (95% CI = 40, 51), followed by a modest 2.4% increase to the end of the study period in April 2023 (95% CI [2.0,2.7])., Conclusions: A 45% reduction in oxycodone consumption in Australia from 2019 to 2020 coincided with national policy changes that aimed to reduce consumption of prescription opioids. The overall declining trend in consumption was suggestive of the effectiveness of national interventions in reducing pharmaceutical opioid use. Wastewater-based epidemiology provides an effective approach for assessing the effectiveness of controlled substances policy changes., (© 2024 The Author(s). Addiction published by John Wiley & Sons Ltd on behalf of Society for the Study of Addiction.)

Tobacco-specific alkaloids and nitrosamines are important biomarkers for the estimation of tobacco use and human exposure to tobacco-specific nitrosamines that can be monitored by wastewater analysis. Thus far their analysis has used solid phase extraction, which is costly and time-consuming. In this study, we developed a direct injection liquid chromatography-tandem mass spectrometry method for the quantification of two tobacco-specific alkaloids and five nitrosamines in wastewater. The method achieved excellent linearity (R 2  > 0.99) for all analytes, with calibration ranging from 0.10 to 800 ng/L. Method limits of detection and quantification were 0.17 ng/L (N-nitrosonornicotine, NNN) and 1.0 ng/L (N-nitrosoanatabine (NAT) and NNN), with acceptable accuracy (100 % ± 20 %) and precision (± 15 %). Analyte loss during filtration was < 15 %, and the relative matrix effect was < 10 %. The method was applied to 43 pooled wastewater samples collected from three wastewater treatment plants in Australia between 2017 and 2021. Anabasine and anatabine were detected in all samples at concentrations of 5.0 - 33 ng/L and 12 - 41 ng/L, respectively. Three of the five tobacco-specific nitrosamines (NAT, NNN, and (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol) (NNAL)) were detected, in < 50 % of the wastewater samples, with concentrations nearly ten times lower than the tobacco alkaloids (< 1.0 - 6.2 ng/L). In-sewer stability of the nitrosamines was also assessed in this study, with four (NAT, NNAL, NNN, and N-nitrosoanabasine (NAB)) being stable (i.e. < 20 % transformation over 12 h in both control reactor (CR) and rising main reactor (RM) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) being moderately stable (< 40 % loss over 12 h in RM). This direct injection method provides a high-throughput approach in simultaneous investigation of tobacco use and assessment of public exposure to tobacco-specific nitrosamines., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Wastewater-based epidemiology (WBE) is proposed as a cost-effective approach to objectively monitor the antidepressant use but it requires more accurate correction factors (CF) than what had been used in previous studies. Amitriptyline is a popular prescription medicine for treating depression and nerve pain, which could be prone to misuse and need monitoring. The CF of amitriptyline employed in previous WBE studies varied from 10 to 100, leading to substantial disparities between WBE estimates and expected mass of antidepressants in wastewater. Hence, this study aimed to take amitriptyline as a case study and refine the CF by correlating mass loads measured in wastewater from 12.2 million inhabitants collected during the 2016 Census with corresponding annual sales data. The triangulation of WBE data and sales data resulted in a newly-derived CF of 7, which is significantly different from the CF values used in previous studies. The newly derived CF was applied to a secondary, multi-year (2017 to 2020) WBE dataset for validation against sales data in the same period, demonstrating the estimated amitriptyline use (380 ± 320 mg/day/1000 inhabitants) is consistent with sales data (450 ± 190 mg/day/1000 inhabitants). When we applied the new CF to previous studies, the wastewater consumption loads matched better to prescription data than previous WBE estimations. The refined CF of amitriptyline can be used in future WBE studies to improve the accuracy of the consumption estimates., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Users of novel psychoactive substances (NPS) are at risk, due to limited information about the toxicity and unpredictable effects of these compounds. Wastewater-based epidemiology (WBE) has been used as a tool to provide insight into NPS use at the population level. To understand the preferences and trends of NPS use in Australia, this study involved liquid chromatography mass spectrometry analysis of wastewater collected from Australian states and territories from February 2022 to February 2023. In total, 59 different NPS were included across two complementary analytical methods and covered up to 57 wastewater catchments over the study. The NPS detected in wastewater were 25-B-NBOMe, buphedrone, 1-benzylpiperazine (BZP), 3-chloromethcathinone, N,N-dimethylpentylone (N,N-DMP), N-ethylheptedrone, N-ethylpentylone, eutylone, 4F-phenibut, 2-fluoro deschloroketamine, hydroxetamine, mephedrone, methoxetamine, methylone, mitragynine, pentylone, phenibut, para-methoxyamphetamine (PMA), alpha-pyrrolidinovalerophenone (α-PVP) and valeryl fentanyl. The detection frequency for these NPS ranged from 3 % to 100 % of the sites analysed. A noticeable decreasing trend in eutylone detection frequency and mass loads was observed whilst simultaneously N,N-DMP and pentylone increased over the study period. The emergence of some NPS in wastewater pre-dates other sources of monitoring and provides further evidence that WBE can be used as an additional early warning system for alerting potential NPS use., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Cobus Gerber reports financial support was provided by Drug and Alcohol Services South Australia. Cobus Gerber reports financial support was provided by Australian Criminal Intelligence Commission., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

One of the primary criteria for a suitable drug biomarker for wastewater-based epidemiology (WBE) is having a unique source representing human metabolism. For WBE studies, this means it is important to identify and monitor metabolites rather than parent drugs, to capture consumption of drugs and not fractions that could be directly disposed. In this study, a high-throughput workflow based on a human liver S9 fraction in vitro metabolism assay was developed to identify human transformation products of new chemicals, using α-pyrrolidino-2-phenylacetophenone (α-D2PV) as a case study. Analysis by liquid chromatography coupled to high resolution mass spectrometry identified four metabolites. Subsequently, a targeted liquid chromatography - tandem mass spectrometry method was developed for their analysis in wastewater samples collected from a music festival in Australia. The successful application of this workflow opens the door for future work to better understand the metabolism of chemicals and their detection and application for wastewater-based epidemiology., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Plastics are ubiquitous in virtually every environment on earth. While the specific sources of plastics entering wastewater are not well known, growing evidence suggests sewage sludge (biosolids) can be a sink for plastics. One potential source could be the sewerage pipe materials used to transport sewage between premises and wastewater treatment plants (WWTPs). To evaluate the significance of sewerage piping as a source of biosolids plastics concentrations, we compared the proportion of the total network (by length and surface area) of polyethylene (PE), polyvinylchloride (PVC), and polypropylene (PP) pipes from 10 WWTPs against their biosolids mass concentrations (mg plastic/g biosolid). Among the 10 catchments, the percentage of the network consisting of PP piping ranged from 0 to 1 %, with 0.8-21 % for PE, and 8-73 % for PVC. Biosolids plastics concentrations ranged from 0.09 to 8.62 mg/g (mg plastic/g biosolid) for PP and PE, respectively. For all three plastics, there was no significant Pearson correlation (r < 0.4) between the biosolids concentration (dry weight mg/g) and the proportion of the network material of the sewerage piping as plastic (either length or surface area). A comparison of trade waste entering a subset of 6 WWTP showed the highest biosolid principal components analysis (PCA) associations between loads of plastics (g/day) and automotive wash bays, general manufacturing, hospitals, laboratories, food manufacturing, laundry and dry cleaning, and cooling towers. A stepwise regression analysis indicated pipe length and surface area, as well as automotive wash bays and food manufacturing may be significant. While our data gave mixed results on the attribution of the sources of plastics entering WWTPs, it suggests that sewerage infrastructure and trade waste may play some role. Future studies should investigate the leachability of sewerage infrastructure and contributions from specific trade waste categories to determine their significance in plastics entering WWTPs., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ben Tscharke reports financial support was provided by Australian Research Council. Elvis Okoffo reports financial support was provided by Australian Research Council. Jiaying Li reports financial support was provided by Australian Research Council. Kevin Thomas reports financial support was provided by Australian Research Council., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Previous wastewater-based epidemiology (WBE) studies have reported decreasing trends of nicotine and tobacco use in Australia before 2017, but there is concern that increasing illicit use of nicotine in vaping products and illicit tobacco could reverse this progress. This study aimed to assess temporal trends of nicotine consumption and specifically tobacco consumption via wastewater analysis in a population in Australia between 2013 and 2021. One week of daily wastewater samples were analyzed every two months from February 2013 to December 2021 in a regional city serving ∼100,000 people. A total of 340 daily samples were analyzed for anabasine (tobacco specific biomarker) and nicotine metabolites, cotinine and hydroxycotinine, using direct injection method by liquid chromatography with tandem mass spectrometry. Daily consumption estimates were calculated from daily flow data, population estimates and previously reported excretion factors. Linear spline regression was performed to identify periods when significant change of slopes occurred and to evaluate the temporal trends. Tobacco use monitored using anabasine as a biomarker, showed a decreasing trend over the whole period with a higher rate of decrease during the first two years (2013-2014, 21 % decrease) compared to the later 7 years (2015-2021, 10 % decrease). Nicotine use, monitored using cotinine and hydroxycotinine, showed a downward trend between 2013 and 2018 (2013-2014: 18 % decrease, p < 0.05; 2015-2016: 6 % increase, p = 0.48; Feb-Dec 2017: 15 % decrease, p = 0.39) followed by a significant increase from 2018 to 2021 (40 % increase, p < 0.001). This finding suggests the increasing use of non-tobacco nicotine-based products. Additionally, the tobacco use estimate by wastewater analysis was higher than the tobacco sales data, which suggests the use of illicit tobacco in the catchment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Pesticides are necessary for the control of pest plant, fungi and insect species. After application, they may find their way into waste streams, such as municipal sewage, where their spatio-temporal distribution has not been well characterised. To further understand the spatio-temporal distribution and to evaluate potential sources and fate after treatment, 64 pesticides were analysed in matched influents and effluents of 22 wastewater treatment plants (WWTPs) from across Australia. The pesticides consisted of 30 herbicides and 8 herbicide metabolites or transformation products, 16 insecticides and 10 fungicides. The samples were 1084 24-hr composite samples pooled into 113 samples. Pools represented two influent and one effluent pools at each of 22 sites in 2019, as well as two pools per year from 2009 to 2021 for an 11-year long-term temporal trend at a subset of two locations. The total population served by the 22 sites was equivalent to ~41 % of the Australian population. Of the 64 pesticides, 25 were detected in influent, with highest influent concentrations up to 100 μg/L and effluent concentrations up to 16 μg/L for the herbicide 2,4-D. The total mass of pesticides was extrapolated to Australia, suggesting ~33 t of the targeted pesticides entered WWTP influent annually nation-wide, with 14 t emitted into effluents annually. Long-term trends varied by analyte and for carbendazim decreases over time, may be related to restrictions in use. Risk quotients (RQs) were calculated for 14 analytes in the effluent. 35 % had an RQ above one, indicating a potential environmental risk. Fipronil had the highest RQ (49) at Site 6. The population-normalized mass loads of pesticides were site-specific, and in some cases correlated with land use attributes suggestive of point sources. This reflects a need to better characterise sources to enable prevention, or possible pre-treatment of pesticide-containing wastewater entering municipal sewage streams., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ben Tscharke reports financial support was provided by Australian Criminal Intelligence Commission. Jochen Mueller reports financial support was provided by Australian Research Council. Emma Knight, Ben Tscharke and Jochen Mueller reports financial support was provided by the Australian Government Department of Agriculture, Fisheries and Forestry., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Wastewater-based epidemiology (WBE) relies on representative sampling that is typically achieved with autosamplers that collect time, flow, or volume proportional samples. The expense, resources and operational know-how associated with autosampler operation means they are only typically available at major wastewater treatment plants (WWTPs). This results in a lack of data on consumption levels in regional and remote areas, or in countries that lack the financial means. The aim of this study was to estimate and investigate trends in drug consumption across varying levels of remoteness in Australia. Field-calibrated, microporous polyethylene passive samplers were deployed over 2 periods (Aug/Sept 2019 and 2020) at 43 treatment plants covering all five categories of remoteness, as per Australian Bureau of Statistics definitions (Major cities, Inner regional, Outer regional, Remote, and Very remote). The per capita consumption of cocaine, methylamphetamine, nicotine, oxycodone and MDMA were estimated. No spatial trends between remoteness and drug consumption were observed, except for cocaine, where Major cities had a 5-to-10-fold higher consumption compared to the other levels of remoteness in 2019 and 2020, respectively. Outer regional sites had the highest and lowest methylamphetamine consumption. The variance in drug use among sites was much higher in Remote (and Inner/Outer regional) sites when compared with Major cities. A significant and consistent decrease in oxycodone consumption was observed at all sites between 2019 and 2020, possibly related to regulatory changes and the COVID-19 pandemic where elective surgeries were suspended. The majority of sites experienced a decrease in cocaine and methylamphetamine consumption, possibly due to border restrictions or changes in supply and demand dynamics. This was the first extensive passive sampling study to assess drug consumption in urban, regional, and remote locations, demonstrating that passive samplers can facilitate extension of wastewater-based drug monitoring programs to sites where other representative sampling options are very difficult to implement., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Tobacco use is known to cause health damage, partly by changing the mouth, respiratory tract, and gut-related microbiomes. This study aims to identify the associations between the human microbiome detected in domestic wastewater and the population smoking rate. Metagenomic sequencing and a biomarker discovery algorithm were employed to identify microorganisms as potential microbial biomarkers of smoking through wastewater-based epidemiology. Wastewater samples were collected from selected catchments with low and high smoking rates, i.e., 11.2 ± 1.5% and 17.0 ± 1.6%, respectively. Using the linear discriminant analysis effect size (LEfSe) method, Neisseria , Desulfovibrio , Megamonas , Blautia , Fusicatenibacter , Granulicatella and Enterococcus were suggested as potential biomarker microorganisms. A higher abundance of pathogens, including Neisseria , Eikenella and Haemophilus , was associated with the high smoking rate, likely because of their colonization in smoking-disturbed human guts. The identified potential microbial biomarkers reflect the change of the human gut microbiome due to the long-term smoking behavior. The metagenomic analysis also indicates that smoking upregulates microbial gene expression of genetic information processing, environmental information processing, and cell wall peptidoglycan cleavage, while it downregulates amino acid, lipid, and galactose metabolisms. The findings demonstrate the potential of microbial biomarkers for the surveillance of smoking through a wastewater-based epidemiology approach., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Co-published by Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, and American Chemical Society.)

Drug consumption in prisons is a concern for the safety of incarcerated people and staff. Typically, drug use prevalence in prisons is estimated through urinalysis and intelligence operations, which can be intrusive and stressful. An alternative approach, wastewater-based epidemiology (WBE), was used in this study to estimate the consumption of licit and illicit drugs for the entire population of a prison in Australia. Wastewater samples were collected from March to December 2020, covering periods of no restrictions and periods when prison access was restricted to prevent the transmission of COVID-19. Target biomarkers were analysed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The average consumption of common illicit drugs (MDMA, methamphetamine and cocaine) over the sampling period in the prison (0.5 - 4.5 mg/1000 people/day) was two to three orders of magnitude lower than in the community population (254 - 1000 mg/1000 people/day). Comparison of WBE estimates against pharmacy dispensing data suggested potential illicit buprenorphine consumption at the prison. Methamphetamine and buprenorphine use decreased when no visitors were allowed (18% - 72% decrease for methamphetamine; about half decrease for buprenorphine) and increased once these restrictions were eased (22% - 39% increase for methamphetamine; 44% - 67% increase for buprenorphine). The changes in drug use may be attributed in part to a reduction of drug trafficking into the prison from visitors or non-essential staffs and in part to the reduced contribution of urine from staff who used toilets within the prison. This study provided useful information on the scale of illicit drug use and extra-medical use of licit drugs in prison, and its changes under different security conditions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

In wastewater-based epidemiology (WBE), nicotine metabolites have been used as biomarkers for monitoring tobacco use. Recently, the minor tobacco alkaloids anabasine and anatabine have been suggested as more specific biomarkers for tobacco use since nicotine use can be from both tobacco and non-tobacco sources. This study aimed to provide an in-depth evaluation of the suitability of anabasine and anatabine as WBE biomarkers of tobacco and subsequently estimate their excretion factors for WBE applications. Pooled urine ( n = 64) and wastewater samples ( n = 277), collected between 2009 and 2019 in Queensland, Australia, were analyzed for nicotine and its metabolites (cotinine and hydroxycotinine), as well as anabasine and anatabine. Anabasine performed as the better biomarker, showing a similar per capita load in pooled urine (2.2 ± 0.3 μg/day/person) and wastewater samples (2.3 ± 0.3 μg/day/person), while the per capita load of anatabine in wastewater was 50% higher than its load in urine. It is estimated that 0.9 μg of anabasine was excreted per cigarette smoked. Triangulation of tobacco sales data and tobacco use estimated from either anabasine or cotinine showed that anabasine-based estimates were 5% higher than sales data, while cotinine-based estimates were between 2 and 28% higher. Our results provided concrete evidence to confirm the suitability of anabasine as a specific biomarker for monitoring tobacco use by WBE.

Introduction: Mixed findings have been reported about the impact of the COVID-19 pandemic on smoking behavior in different populations., Aims and Methods: In this study, we aimed to quantify changes in smoking prevalence through the proxy of nicotine consumption in the Australian population from 2017 to 2020 inclusive. Estimates of nicotine consumption between 2017 and 2020 were retrieved from a national wastewater monitoring program that covers up to 50% of the Australian population. National sales data for nicotine replacement therapy (NRT) products from 2017 to 2020 were also acquired. Linear regression and pairwise comparison were conducted to identify data trends and to test differences between time periods., Results: The average consumption of nicotine in Australia decreased between 2017 and 2019 but increased in 2020. Estimated consumption in the first half of 2020 was significantly higher (~30%) than the previous period. Sales of NRT products increased gradually from 2017 to 2020 although sales in the first half of the year were consistently lower than in the second half., Conclusion: Total nicotine consumption increased in Australia during the early stage of the pandemic in 2020. Increased nicotine consumption may be due to people managing higher stress levels, such as from loneliness due to control measures, and also greater opportunities to smoke/vape while working from home and during lockdowns in the early stage of the pandemic., Implications: Tobacco and nicotine consumption have been decreasing in Australia but the COVID-19 pandemic may have temporarily disrupted this trend. In 2020, the higher impacts of lockdowns and working from home arrangements may have led to a temporary reversal of the previous downward trend in smoking during the early stage of the pandemic., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Research on Nicotine and Tobacco.)

Background and Aim: From 1 February 2018, codeine was rescheduled from an over-the-counter (OTC) to a prescription-only medicine in Australia. We used wastewater-based epidemiology to measure changes in population codeine consumption before and after rescheduling., Methods: We analysed 3703 wastewater samples from 48 wastewater treatment plants, taken between August 2016 and August 2019. Our samples represented 10.6 million people, 45% of the Australian population in state capitals and regional areas in each state or territory. Codeine concentrations were determined by liquid chromatography-tandem mass spectrometry and converted to per-capita consumption estimates using the site daily wastewater volume, catchment populations and codeine excretion kinetics., Results: Average per-capita consumption of codeine decreased by 37% nationally immediately after the rescheduling in February 2018 [95% confidence interval (CI) = 35.3-39.4%] and substantially in all states between 24 and 51% (95% CI = 22.4-27.0% and 41.8-59.4%). The decrease was sustained at the lower level to August 2019. Locations with least pharmacy access decreased by 51% (95% CI = 41.7-61.7%), a greater decrease than 37% observed for those with greater pharmacy access (95% CI = 35.1-39.4%). Regional areas decreased by a smaller margin to cities (32 versus 38%, 95% CI = 30.2-34.1% versus 34.9-40.4%, respectively) from a base per-capita usage approximately 40% higher than cities., Conclusion: Wastewater analysis shows that codeine consumption in Australia decreased by approximately 37% following its rescheduling as a prescription-only medicine in 2018. Wastewater-based epidemiology can be used to evaluate changes in population pharmaceutical consumption in responses to changes in drug scheduling., (© 2022 The Authors. Addiction published by John Wiley & Sons Ltd on behalf of Society for the Study of Addiction.)

Consumption of amphetamine and methamphetamine, two common illicit drugs, has been monitored by wastewater-based epidemiology (WBE) in many countries over the past decade. There is potential for the estimated amount of amphetamine used to be skewed at locations where methamphetamine is also consumed, because amphetamine is also excreted to wastewater following methamphetamine consumption. The present study aims to review the available data in the literature to identify an average ratio of amphetamine/methamphetamine (AMP/METH) that is excreted to wastewater after methamphetamine consumption. This ratio could then be used to refine the estimation of amphetamine consumption in catchments where there is both amphetamine and methamphetamine use. Using data from more than 6000 wastewater samples from Australia where methamphetamine is the dominant illicit amphetamine-type substance on the market, we were able to subtract the contribution of legal sources of amphetamine contribution and obtain the median AMP/METH ratio in wastewater of 0.09. Using this value, the amphetamine derived from methamphetamine consumption can be calculated and subtracted from the total amphetamine mass loads in wastewater samples. Without considering the contribution of amphetamine from methamphetamine use, selected European catchments with comparable consumption of amphetamine and methamphetamine showed up to 83% overestimation of amphetamine use. For catchments with AMP/METH ratio greater than 1.00, the impact of amphetamine from methamphetamine would be negligible; for catchments with AMP/METH ratio in the range of 0.04-0.19, it will be difficult to accurately estimate amphetamine consumption., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

Analysis of untreated municipal wastewater is recognized as an innovative approach to assess population exposure to or consumption of various substances. Currently, there are no published wastewater-based studies investigating the relationships between catchment social, demographic, and economic characteristics with chemicals using advanced non-targeted techniques. In this study, fifteen wastewater samples covering 27% of the Australian population were collected during a population Census. The samples were analysed with a workflow employing liquid chromatography high-resolution mass spectrometry and chemometric tools for non-target analysis. Socioeconomic characteristics of catchment areas were generated using Geospatial Information Systems software. Potential correlations were explored between pseudo-mass loads of the identified compounds and socioeconomic and demographic descriptors of the wastewater catchments derived from Census data. Markers of public health (e.g., cardiac arrhythmia, cardiovascular disease, anxiety disorder and type 2 diabetes) were identified in the wastewater samples by the proposed workflow. They were positively correlated with descriptors of disadvantage in education, occupation, marital status and income, and negatively correlated with descriptors of advantage in education and occupation. In addition, markers of polypropylene glycol (PPG) and polyethylene glycol (PEG) related compounds were positively correlated with housing and occupation disadvantage. High positive correlations were found between separated and divorced people and specific drugs used to treat cardiac arrhythmia, cardiovascular disease, and depression. Our robust non-targeted methodology in combination with Census data can identify relationships between biomarkers of public health, human behaviour and lifestyle and socio-demographics of whole populations. Furthermore, it can identify specific areas and socioeconomic groups that may need more assistance than others for public health issues. This approach complements important public health information and enables large-scale national coverage with a relatively small number of samples., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Opioids are commonly used as analgesics to relieve chronic pain and have high abuse potential. Due to their strong potency and trace concentration in plasma, a robust analytical method is necessary for quantification in forensic and pharmacology fields. Hence, this study developed and validated a simple, rapid, and robust method for the simultaneous determination of 12 opioids and metabolites which were available legally by prescription or abused for non-medical purposes, in plasma samples by simple liquid extraction and high-performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). We compared the extraction recovery of our sample pre-treatment to two other sample pre-treatments (namely QuEChERS and simplified QuEChERS) and showed that the method used in our study gave the highest recoveries. The method validation followed the European Medicines Agency guidelines, including selectivity, carryover, accuracy and precision, dilution integrity, matrix effect and freeze/thaw stability. This method's accuracy ranged from 85% to 115% with a precision less than 15%, within the acceptable range of the validation protocol. The lower limit of quantification of the method ranged between 0.05 μg L -1 and 0.38 μg L -1 among 12 opioids/metabolites. Stability was assessed, with all opioids observed as relatively stable at 0.5 μg L -1 and 5 μg L -1 levels under -20 °C and 25 °C storage conditions. In summary, the developed method has the potential to achieve simultaneous analysis for monitoring opioids in forensic and pain management regimens using a simple sample pre-treatment.

This study investigated the occurrence and contribution of plastic particles associated with size fractionated biosolids to the total concentration in biosolids (treated sewage sludge) samples collected from 20 wastewater treatment plants (WWTP) across Australia. This was achieved through sequential size fractionation of biosolids samples to quantify the mass concentration of 7 common plastics across a range of biosolids size fractions, including below 25 μm which has not been assessed in many previous studies. Quantitative analysis was performed by pressurized liquid extraction followed by pyrolysis coupled to gas chromatography - mass spectrometry. Of the total quantified plastics (Σ 7 plastics), the greatest proportion (27%) of the total mass were identified in the nominal <25 μm sized biosolids fraction. Polyethylene dominated the polymer mass in every size fraction, even though profiles varied between WWTPs. When comparing the sum of all sites for each sized biosolids fraction, the plurality of the polyethylene, polyvinyl-chloride, polystyrene, polypropylene, polycarbonate, and polyethylene-terephthalate concentrations were associated with the smallest size fraction (<25 μm). We confirm for the first time the presence of plastic particles in biosolids below a size fraction that is not captured by many methods. This is important, because of the potential greater significance of plastics in the low sizes to environmental and human health., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Wastewater-based epidemiology is a potential complementary technique for monitoring the use of performance- and image-enhancing drugs (PIEDs), such as anabolic steroids and selective androgen receptor modulators (SARMs), within the general population. Assessing in-sewer transformation and degradation is critical for understanding uncertainties associated with wastewater analysis. An electrospray ionization liquid chromatography mass spectrometry method for the quantification of 59 anabolic agents in wastewater influent was developed. Limits of detection and limits of quantification ranged from 0.004 to 1.56 μg/L and 0.01 to 4.75 μg/L, respectively. Method performance was acceptable for linearity ( R 2 > 0.995, few exceptions), accuracy (68-119%), and precision (1-21%RSD), and applicability was successfully demonstrated. To assess the stability of the selected biomarkers in wastewater, we used laboratory-scale sewer reactors to subject the anabolic agents to simulated realistic sewer environments for 12 h. Anabolic agents, including parent compounds and metabolites, were spiked into freshly collected wastewater that was then fed into three sewer reactor types: control sewer (no biofilm), gravity sewer (aerobic conditions), and rising main sewer (anaerobic conditions). Our results revealed that while most glucuronide conjugates were completely transformed following 12 h in the sewer reactors, 50% of the investigated biomarkers had half-lives longer than 4 h (mean residence time) under gravity sewer conditions. Most anabolic agents were likely subject to biofilm sorption and desorption. These novel results lay the groundwork for any future wastewater-based epidemiology research involving anabolic steroids and SARMs.

Aim: To test if there was a reduction in alcohol consumption in wastewater samples in the Northern Territory of Australia after the implementation of a minimum unit alcohol price policy (MUP) in October 2018., Design, Setting, Cases: Between August 2016 and February 2020, wastewater samples were collected across 66 sites in the Northern Territory and all other states and territories in Australia. Samples were collected every 2 months in capital cities and every 4 months in regional places during this period. Overall, 4917 samples were taken (2816 before MUP and 2101 after)., Measurements: The number of standard drinks per 1000 people per day in the respective catchment areas was estimated based on the concentration of an alcohol-specific metabolite, ethyl sulphate in the samples (using the excretion factor of ethyl sulphate, the flow of wastewater entering the wastewater treatment plants and the population of each wastewater catchment)., Findings: Results from a linear mixed model showed that there was a large drop in alcohol consumption immediately after the MUP in Northern Territory [estimated drop = 1231, 99% confidence interval (CI) = 830, 1633; 38.75%]. There was no significant drop in all other states/territories except for Queensland, which showed a significant but much smaller drop (estimated drop: 310; 99% CI = 114, 550). One year after the MUP, the drop narrowed to 520 (99% CI = 189, 851) and was no longer statistically significant in February 2020 (15 months after MUP; estimated drop = 283, 99% CI = -114, 681)., Conclusions: Per-capita consumption of alcohol appears to have decreased substantially in the Northern Territory of Australia immediately after the implementation of a minimum unit price but consumption steadily recovered and almost returned to the pre-MUP consumption level after 15 months., (© 2021 Society for the Study of Addiction.)

Performance- and image-enhancing drug (PIED) misuse is a significant public health issue. Currently, seizure data, surveys, anti-doping testing, and needle service provider data are used to estimate PIED use in populations. These methods are time consuming, single point-in-time measurements that often consist of small sample sizes and do not truly capture PIED prevalence. Wastewater-based epidemiology (WBE) has been used globally to assess and monitor licit and illicit drug consumption within the general community. This method can objectively cover large populations as well as specific subpopulations (gyms, music festivals, prisons), and has potential as a complementary monitoring method for PIED use. Information obtained through WBE could be used to aid public health authorities in developing targeted prevention and education programmes. Research on PIED analysis in wastewater is limited and presents a significant gap in the literature. The focus is on anabolic steroids, and one steroid alternative currently growing in popularity; selective androgenic receptor modulators. This encompasses medical uses, addiction, prevalence, user typology, and associated public health implications. An overview of WBE is described including its benefits, limitations and potential as a monitoring method for PIED use. A summary of previous work in this field is presented. Finally, we summarise gaps in the literature, future perspectives, and recommendations for monitoring PIEDs in wastewater., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Passive sampling approaches to monitor licit and illicit drugs of concern in wastewater shows promise as a supplementary sampling technique to grab sampling when conventional composite autosampling may not be possible. Recent studies have assessed the applicability of passive sampling at a single wastewater treatment plant (WWTP). However, it remains unknown whether a single-site calibration can be applied to other WWTPs. This study evaluated the in situ calibration of microporous polyethylene tube passive samplers (MPTs) against simultaneously collected composite samples for 22 different WWTPs. Samples were analyzed for methylamphetamine, amphetamine, hydroxycotinine, cotinine, benzoylecgonine, 3,4-methylenedioxymethamphetamine, and noroxycodone. Estimated rates of chemical uptake (sampling rates) were calculated using the mass accumulated in the samplers, the concentration measured in composite samples, and the duration of deployment. The estimated sampling rates were consistent between WWTPs (>90% within a factor of two) and ranged from 5 mL day -1 (amphetamine) to 9 mL day -1 (noroxycodone). The samplers were effective at estimating analyte concentrations, with 77% of results having a normalized difference to 24 h composite samples of below 30%. Our study suggests that MPT passive samplers provide a tool for the spatiotemporal monitoring of drug use where automated integrative sampling techniques may not be feasible.

Understanding the in-sewer stability of analgesic biomarkers is important for interpreting wastewater-based epidemiology (WBE) data to estimate community-wide analgesic drugs consumption. The in-sewer stability of a suite of 19 analgesics and their metabolites was assessed using lab-scale sewer reactors. Target biomarkers were spiked into wastewater circulating in simulated gravity, rising main and control (no biofilm) sewer reactors. In-sewer transformation was observed over a hydraulic retention time of 12 h. All investigated biomarkers were stable under control reactor conditions. In gravity sewer conditions, diclofenac, desmetramadol, ibuprofen carboxylic acid, ketoprofen, lidocaine and tapentadol were highly stable (0-20% transformation in 12 h). Valdecoxib, parecoxib, etoricoxib, indomethacin, naltrexone, naloxone, piroxicam, ketoprofen, lidocaine, tapentadol, oxymorphone, hydrocodone, meperidine, hydromorphone were considered as moderately stable biomarkers (20-50% transformation in 12 h). Celecoxib and sulindac were considered unstable biomarkers (>50% transformation in 12 h). Ketoprofen, lidocaine, tapentadol, meperidine, hydromorphone were transformed to 0-20% whereas diclofenac, desmetramadol, ibuprofen carboxylic acid, valdecoxib, parecoxib, etoricoxib, indomethacin, naltrexone, piroxicam were transformed up to 20-50% in 12 h in rising main reactor (RMR). These biomarkers were considered as highly stable and stable biomarkers in RMR, respectively. Sulindac, celecoxib, naloxone, oxymorphone and hydrocodone were transformed more than 50% in 12 h and considered as unstable biomarkers in RMR. This study provides the information for a better understanding of the in-sewer loss of the analgesics before using them in WBE biomarkers for estimating drug loads at the population level., (Copyright © 2021. Published by Elsevier Ltd.)

Methamphetamine, MDMA, cocaine, cannabis, and alcohol in samples from 20 wastewater treatment plants servicing the eight state or territory capitals of Australia were analyzed, with equivalent coverage of >45% of the national population. Trends in drug consumption were calculated and assessed from samples collected from 2016 to 2020, with a focus on pre-COVID-19 (August 2016 to December 2019), versus February to June 2020, when Australia observed a nationwide lockdown. Results showed delayed but significant decreases in methamphetamine, >50% in Western Australia. In contrast, significant increases in cannabis in most jurisdictions were observed. This suggests changes in consumption may be somewhat linked to reduced supply of imported substances, with increased use of locally produced drugs. Initial decreases in cocaine and MDMA consumption were evident in many parts of the country, but pre-COVID trends were re-established after April 2020. Interestingly, weekend-weekday differences were narrowed for cocaine, MDMA, and alcohol during lockdown, which might be expected due to bars being closed and social gathering not allowed. With this study providing insight into the first four months of COVID-19 restrictions in Australia, it remains to be seen what the longer-term effect of the pandemic will be., Competing Interests: The authors declare no competing financial interest., (© 2021 American Chemical Society.)

Plastics are ubiquitous contaminants that leak into the environment from multiple pathways including the use of treated sewage sludge (biosolids). Seven common plastics (polymers) were quantified in the solid fraction of archived biosolids samples from Australia and the United Kingdom from between 1950 and 2016. Six plastics were detected, with increasing concentrations observed over time for each plastic. Biosolids plastic concentrations correlated with plastic production estimates, implying a potential link between plastics production, consumption and leakage into the environment. Prior to the 1990s, the leakage of plastics into biosolids was limited except for polystyrene. Increased leakage was observed from the 1990s onwards; potentially driven by increased consumption of polyethylene, polyethylene terephthalate and polyvinyl chloride. We show that looking back in time along specific plastic pollution pathways may help unravel the potential sources of plastics leakage into the environment and provide quantitative evidence to support the development of source control interventions or regulations., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Artificial sweeteners are contaminants of emerging concern that can enter the aquatic and terrestrial environments via wastewater effluent discharge and the environmental application of biosolids. The release of artificial sweeteners from the use of biosolids in Australia was assessed. The concentration of seven artificial sweeteners was quantified in biosolids samples collected from 71 wastewater treatment plants (WWTPs) across Australia during Census 2016. Sucralose, saccharin, acesulfame, aspartame and cyclamate were detected in biosolids samples at median concentrations ranging from 0.18 ng/g (dry weight) (range: -1  person -1 for cyclamate to 4,000 (1,000, 15,000) µg year -1  person -1 for sucralose with 223 kg of artificial sweeteners released to terrestrial environment from biosolids end-use annually in Australia. Due to the low loads of artificial sweeteners in biosolids compared with wastewater effluent, risks associated with artificial sweeteners in biosolids are likely limited., (Copyright © 2021. Published by Elsevier Ltd.)

Aims: A discrete experiment in wastewater-based epidemiology (WBE) timed to coincide with the census was used to investigate the spatial, temporal and socioeconomic patterns of illicit drug consumption in Auckland, Bay of Plenty and Canterbury., Methods: For seven consecutive days over census week (6 March 2018), wastewater was sampled from seven wastewater treatment plants and analysed for methamphetamine, cocaine (as benzoylecgonine) and 3,4-methylenedioxymethamphetamine (MDMA). Detailed sewer catchment maps were developed and, together with the data, were used to analyse drug consumption., Results: Methamphetamine (mean 22.9 ± 9.9 doses/day/1000 people) was the most consumed drug, followed by MDMA (mean 1.7 ± 1.5 doses/day/1000 people) and cocaine (mean 0.5 ± 0.3 doses/day/1000 people). Methamphetamine consumption (and to a lesser extent MDMA) was high compared to that reported for Western nations, while cocaine consumption was extremely low. Cocaine and MDMA consumption were higher in cities compared to towns. In contrast, methamphetamine was typically higher in towns. Cocaine and MDMA were consumed more at weekends. Methamphetamine use was more consistent throughout the week. MDMA and cocaine were correlated with socioeconomic advantage, whereas methamphetamine was correlated with disadvantage., Conclusions: This paper contextualises illicit drug use in three New Zealand regions containing 18.3% of the national population and confirms the pervasiveness of methamphetamine consumption in New Zealand towns. This work demonstrates how WBE can be used to explore the socioeconomic dimensions of drug use when duly combined with other data sources like censuses., Competing Interests: Nil.

The in-sample stability of selected pharmaceuticals, illicit drugs, and their metabolites in wastewater was assessed under six different conditions-untreated, addition of hydrochloric acid or sodium metabisulfite solution, combined with or without sterile filtration, and at four representative temperatures, at 35 °C for up to 28 days, 22 °C for 56 days, and 4 °C and -20 °C for 196 days, or freeze/thaw cycles for 24 weeks. Paracetamol, 6-monoacetylmorphine, morphine, and cocaine were poorly stable in untreated wastewater-e.g., with 50% transformation within 1.2-8.1 days at 22 °C, and acidification reduced their in-sample transformations. Acesulfame, carbamazepine, cotinine, methamphetamine, 3,4-methylenedioxy-methamphetamine (MDMA), ketamine, norfentanyl, 3,4-methylenedioxy- N -ethylamphetamine (MDEA), and norbuprenorphine were highly or moderately stable over the observed period, even in untreated wastewater. Fitting of pseudo-first-order kinetics and the Arrhenius equation was used to develop a multistage transformation estimation model combined with an interactive tool to evaluate possible transformation scenarios of selected biomarkers for the processes from sampling to preanalysis. However, as the wastewater composition can vary between sites and over time, the variability of in-sample stability requires further exploration.

The correction factor (CF) is a critical parameter in wastewater-based epidemiology (WBE) that significantly influences the accuracy of the final consumption estimates. However, most CFs have been derived from a few old pharmacokinetic studies and should be re-evaluated and refined to improve the accuracy of the WBE approach. This study aimed to review and estimate the CFs for atenolol, carbamazepine, and naproxen for WBE using the daily mass loads of those pharmaceuticals in wastewater and their corresponding dispensed prescription data in Australia. Influent wastewater samples were collected from wastewater treatment plants serving approximately 24% of the Australian population and annual national dispensed prescription data. The estimated CFs for atenolol and carbamazepine are 1.37 (95% CI: 1.17-1.66) and 8.69 (95% CI: 7.66-10.03), respectively. Due to significant over-the-counter sales of naproxen, a reliable CF could not be estimated based on prescription statistics. Using an independent dataset of 186 and 149 wastewater samples collected in an urban catchment in 2011 and 2012, WBE results calculated using the new CFs matched well with the dispensed data for atenolol and carbamazepine in the catchment area.

Wastewater-based epidemiology studies use catchment populations to normalise chemical marker mass loads in 24-h composite wastewater samples. However, one of the biggest uncertainties within the field is the accuracy of the population used. A population marker in wastewater may significantly reduce the uncertainty. This study evaluated the catecholamine metabolites - homovanillic acid (HVA) and vanillylmandelic acid (VMA) - as potential population biomarkers. Influent wastewater 24-h composite samples were collected from 38 wastewater catchments from around Australia (representing ~33% of Australia's population), extracted and analysed by liquid chromatography tandem mass spectrometry. Measured mass loads were compared to population sizes determined by mapping catchment maps against high-resolution census data. Both biomarkers correlated with coefficient of determinations (r 2 ) of 0.908 and 0.922 for HVA and VMA, respectively. From the regression analysis, a slope (i.e. the daily per-capita excretion) of 1.241 and 1.067 mg.day -1 .person -1 was obtained for HVA and VMA, respectively. The mass load ratio between VMA:HVA were very similar to that reported in literature for urinary analysis among all catchments. Overall, this study provided further evidence that catecholamine metabolites are suitable candidates as population biomarkers for future studies., Competing Interests: Declaration of competing interest The authorshave no affiliation with any organization with a direct or indirect financial interest in the subject matter discussed in the manuscript, (Copyright © 2020 Elsevier B.V. All rights reserved.)

Increased production and use of plastics has resulted in growth in the amount of plastic debris accumulating in the environment, potentially fragmenting into smaller pieces. Fragments <5 mm are typically defined as microplastics, while fragments <0.1 μm are defined as nanoplastics. Over the past decade, an increasing number of studies have reported the occurrence and potential hazards of plastic particles in the aquatic environment. However, less is understood about plastic particles in the terrestrial environment and specifically how much plastic accumulates in soils, the possible sources, potential ecological impacts, interaction of plastic particles with the soil environment, and appropriate extraction and analytical techniques for assessing the above. In this review, a comprehensive overview and a critical perspective on the current state of knowledge on plastic pollution in the soil environment is provided: detailing known sources, occurrence and distribution, analytical techniques used for identification and quantification and the ecological impacts of particles on soil. In addition, knowledge gaps are identified along with suggestions for future research.

A 7-year temporal trend study of artificial sweetener consumption was performed by determining per capital mass loads in 293 influent wastewater samples collected from a wastewater treatment plant in Australia between 2012 and 2018. Population-weighted per capita mass loads of the four detected artificial sweeteners ranged from 2.4 ± 0.8 mg d -1 p -1 for saccharin to 7.8 ± 2.0 mg d -1 p -1 for acesulfame over the study period. Negligible intra-week fluctuations were observed, however the consumption of acesulfame was seen to be significantly influenced by season with the highest consumption in summer. The consumption of sucralose and saccharin significantly increased with an annual increase rate of 10% and 6.0%. Cyclamate consumption declined over the same period with average annual decrease rate of 11%, which agrees with data from market surveys. Sucrose equivalence of total artificial sweeteners consumption showed an increase between 2012 and 2016, then decreased in 2018. This is the first long-term trend study of artificial sweetener consumption by wastewater analysis and highlights the feasibility to quantitatively measure artificial sweeter consumption over time., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Measurement of population tobacco use via wastewater-based epidemiology (WBE) provides objective data to evaluate the efficacy of tobacco control strategies. However, current WBE tobacco-use estimates based on nicotine metabolites (cotinine and hydroxycotinine) can be masked by use of non-tobacco nicotine-containing products. To better understand nicotine and tobacco use, we analysed tobacco-specific biomarkers, anabasine and anatabine, as well as nicotine metabolites, cotinine and hydroxycotinine, in wastewater samples collected for 6 weeks per year over 6 years (2012-2017) from an Australian wastewater treatment plant serving approximately 100,000 people. Population-normalised mass loads were used to estimate tobacco and nicotine use trends and were compared with surveys and taxation statistics. Significant annual declines were observed for anabasine, anatabine, cotinine and hydroxycotinine of -3.0%, -2.7%, -2.4%, and -2.1%, respectively. The results corresponded with the annual declining trends reported from surveys (-5%) and taxation statistics (-4%). Significant annual decreases in the ratios of anabasine to cotinine (-1.2%) and anatabine to cotinine (-1.0%) suggested a relative increase in the use of non-tobacco nicotine products at the same time that tobacco use was declining. Monitoring tobacco use with anabasine and anatabine removed influence from nicotine-containing products, showing larger reductions in this Australian city than via nicotine biomarkers, whilst also demonstrating their suitability for monitoring long-term trends., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Plastics are contaminants of emerging concern that can enter the environment from multiple sources, including via land application of treated sewage sludge (biosolids). Biosolids samples collected from 82 wastewater treatment plants (WWTPs) across Australia and covering 34% of the population during census week in 2016 were quantitatively analyzed to estimate the release of seven common plastics. Quantitative analysis was performed by pressurized liquid extraction followed by double-shot microfurnace pyrolysis coupled to gas chromatography mass spectrometry. Ninety nine percent of the samples contained plastics (Σ 6 plastics) at concentrations of between 0.4 and 23.5 mg/g dry weight (median; 10.4 mg/g dry weight), while polycarbonate was not detected in any sample. Per-capita mass loads of plastics (Σ 6 plastics) released were between 8 and 877 g/person/year across all investigated WWTPs. Polyethylene was the predominant plastic detected, contributing to 69% of Σ 6 plastics. Based on the concentrations measured, it was projected that around 4700 metric tons (Mt) of plastics are released into the Australian environment through biosolids end-use each year, equating to approximately 200 g/person/year, which represents 0.13% of total plastics use in Australia. Of this, 3700 Mt of plastics are released to agricultural lands and 140 Mt to landscape topsoil. Our results provide a first quantitative per-capita mass loads and emission estimate of plastic types through biosolids end-use.

Wastewater-based epidemiology (WBE) has been used to estimate tobacco use in the population. However, the increased use of nicotine replacement therapies and e-cigarettes contributes to the load of nicotine metabolites in wastewater, causing over-estimation of tobacco use if nicotine metabolites were used in WBE back-estimation. This study aims to develop a rapid method for determining the tobacco-specific biomarkers, anabasine and anatabine, in wastewater and to evaluate their in-sewer stability for better estimation of tobacco use by WBE. An enhanced direct injection LC-MS/MS was developed to quantify anabasine and anatabine as well as nicotine biomarkers (nicotine, cotinine and hydroxycotinine). The method was optimal when wastewater was filtered through 0.2 μm RC syringe filters and a pre-conditioned SPE cartridge (Oasis HLB 1 cc, 30 mg) before 50 μL was injected into the LC-MS/MS system. Limits of quantification varied between 2.7 and 54.9 ng/L with recoveries from 76% to 103% for all five compounds. In sewer reactors, anabasine and anatabine were less stable than cotinine and hydroxycotinine. They were more stable in the gravity sewer reactor with <20% loss in 12 h than in the rising main sewer reactor with ~30% loss in the same period. We then applied the new method to 42 daily wastewater influent samples collected from an Australian wastewater treatment plant. The five biomarkers were detected in all samples with concentrations ranging from 9.2 to 7430 ng/L. All five compounds were positively correlated with one another. Our results suggested a high throughput analytical method for feasible application in anabasine and anatabine as biomarkers of tobacco use in routine wastewater monitoring., Competing Interests: Declaration of competing interest Authors declare no competing interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Artificial sweeteners are used as sugar substitutes in our daily lives yet consumption and release patterns are currently unknown in Australia. The spatial distribution of artificial sweetener consumption and WWTP effluent emission in Australia was estimated by wastewater analysis. Wastewater influent and effluent samples were collected from 69 WWTPs across Australia during the week of the 2016 Australian census. Mean population-weighted per capita loads for individual artificial sweeteners (cyclamate, aspartame, acesulfame, sucralose, saccharin) ranged from 0.12 ± 0.14 mg d -1 p -1 for aspartame to 6.9 ± 2.8 mg d -1 p -1 for acesulfame with 1004 kg of these artificial sweeteners being consumed daily in Australia. Significant removal of aspartame (100%), cyclamate (92 ± 18%) and saccharin (88 ± 21%) was observed during wastewater treatment. The average per capita release to the environment for individual artificial sweeteners (cyclamate, acesulfame, sucralose, saccharin) ranged from 230 ± 780 mg d -1 1000p -1 (cyclamate) to 3800 ± 1400 mg d -1 1000p -1 (sucralose). The daily release of artificial sweeteners from Australian WWTPs was estimated to be 142 kg suggesting that 14% of the artificial sweeteners consumed in Australia are released into the environment. To the best of our knowledge, this is the first wastewater study to estimate the occurrence and population-normalized artificial sweetener consumption and emission in Australia., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Owing to the risks and dangers of xenobiotic cannabinoids (phyto and synthetic), studies are required to evaluate community consumption. The analysis of excreted cannabinoids in wastewater can provide information about community consumption for a defined catchment area. The recovery of cannabinoids from complex wastewater matrices is difficult due to the hydrophobic properties of these compounds. In this study, a liquid chromatography tandem mass spectrometry (LC-MS/MS) method was optimised for the recovery of 30 cannabinoids from wastewater, including the cannabis urinary biomarker 11-nor-9-carboxy-Δ 9 -tetrahydrocannabinol (THC-COOH), cannabidiol (CBD), and a variety of different generational synthetic cannabinoids and metabolites. Method validation assessed criteria including linearity, selectivity, recovery, ion-suppression, filtration losses and matrix effect. Two sample preparation approaches-liquid-liquid extraction (LLE) and solid phase extraction (SPE)-were compared, with comparable limits of quantification between 0.001 and 0.5 µg L -1 in wastewater. Filtration was found to reduce the recovery for many of the investigated cannabinoids, where up to 97% of analyte was lost. The method was applied to 15 different catchment areas across Australia to gauge the community use of the cannabinoids in this study. The cannabis biomarker THC-COOH was quantified at all locations, and cannabidiol was measured at eight catchment areas. Three synthetic cannabinoids were detected at the limit of detection: 5-fluoro APINACA, JWH-073 (4-hydroxypentyl), and MDMB-CHMICA., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Community tobacco use can be monitored over time using wastewater-based epidemiological approaches by estimating the mass loads of nicotine and its metabolites, cotinine, or hydroxycotinine, in wastewater. However, due to the use of nicotine in smoking cessation products, other sources of nicotine contribute to cotinine and hydroxycotinine loads. The use of nicotine replacement therapies could vary in space and time and mask the true rates of tobacco consumption. Therefore, this work evaluated the content of tobacco specific markers, anatabine and anabasine, in cigarettes, in urine of smokers, and in wastewater. The results indicated that the anabasine content in both licit and illicit cigarettes in Australia is less variable than anatabine and is therefore considered a better measure of tobacco consumption. A study determining the excretion of tobacco-specific alkaloids of smoking and non-smoking volunteers gave an average urinary mass load of anabasine of 4.38 μg/L/person and a daily mass load of 1.13 μg/day/person. Finally, this was compared with the mass loads of anabasine from wastewater-based epidemiology data of 3 μg/day/person to estimate cigarette rates in a South Australian city: equivalent to 2.6 cigarettes/person/day. The rate of decline of cigarette use was greater when using anabasine as a measure of consumption compared with cotinine. This is the first study to estimate the rate of anabasine excretion, which can be used to estimate tobacco use independent of therapeutically prescribed nicotine., (© 2020 John Wiley & Sons, Ltd.)

Measuring community consumption of new psychoactive substances (NPS) is notoriously difficult to assess by traditional means such as surveys and seizure data. Previously, we used the approach to demonstrate the prevalence of NPS on a national scale. In the current study we explored the temporal resolution for the analysis of NPS in wastewater from Australia. Samples covering all States and Territories in Australia and both metropolitan and regional areas and were collected bimonthly from October 2017-June 2018 and October 2019-February 2020. A qualitative screening method was applied, screening for 201 NPS. In total, 15 substances were found from a variety of classes of NPS. The most prevalent class was synthetic cathinones, with pentylone, N-ethylpentylone and ethylone found in all periods in at least one site in the earlier sampling period, as well as the amphetamine-like paramethoxyamphetamine (PMA). In the latter period, synthetic cathinones were also the most prevalent, including eutylone, marking the first time that this compound has been detected in wastewater. This study shows the application of wastewater analysis to detect outbreaks of NPS use and temporal differences among sites., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Health concerns and related regulation of bisphenol A (BPA) in some countries have led to an increase in the production and use of unregulated and poorly understood BPA analogues, including bisphenol S (BPS), bisphenol F (BPF), bisphenol B (BPB), and bisphenol AF (BPAF). To assess the temporal trends of human exposure to BPA analogues, urine and wastewater samples were collected from South East Queensland, Australia between 2012 and 2017 and analyzed for five bisphenols using validated isotope dilution liquid chromatography tandem mass spectrometry methods. BPA and BPS were the predominant bisphenols detected in both urine and wastewater samples, with median concentrations of 2.5 and 0.64 μg/L in urine and 0.94 and 1.1 μg/L in wastewater, respectively. BPB, BPF, and BPAF had low detection frequencies in both urine and wastewater samples. Concentrations of BPA in both urine and wastewater decreased over the sampling period, whereas concentrations of BPS increased, suggesting that BPS has become a BPA replacement. The contributions of urinary excretion to wastewater were calculated by the ratio of daily per capita urinary excretion to wastewater-based mass loads of bisphenols. Urinary BPA and BPS contributed to less than 1% of the load found in wastewater, indicating that much of the BPA and BPS originates from other sources.

The identification and quantification of selected plastics (polystyrene (PS), polycarbonate (PC), poly-(methyl methacrylate) (PMMA), polypropylene (PP), polyethylene terephthalate (PET), polyethylene (PE) and polyvinyl chloride (PVC)) in biosolids (treated sewage sludge) was performed by pressurized liquid extraction (PLE) combined with double-shot pyrolysis gas chromatography-mass spectrometry. Validation of the method yielded recoveries of between 85 and 128% (mean RSD 11%) at a linear range of between 0.01 and 2 μg. The distribution of plastics within 25 biosolid samples from a single wastewater treatment plant in Australia was assessed. The mass concentration of PE, PVC, PP, PS and PMMA was between 0.1 and 4.1 mg/g dry weight (dw) across all samples, with a total plastic concentration Ʃ Plastics of between 2.8 and 6.6 mg/g dw (median = 4.1 mg/g dw). PE was the predominant plastic detected (mean concentration of 2.2 mg/g dw), contributing to 50% of the total of all plastics. Overall, this study demonstrates that pressurized liquid extraction (PLE) combined with double-shot pyrolysis gas chromatography-mass spectrometry can be used to identify and quantify PE, PP, PVC, PS, and PMMA in biosolids., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Aims: To compare long-term trends in wastewater data with other indicators of stimulant use in three locations and to test the reliability of estimates based on 1 week of sampling., Design: Comparison of trends in quantities ('loads') of stimulants or their metabolites in wastewater with trends in other indicators of stimulant use (e.g. treatment, police, population survey data)., Setting and Participants: Populations in Oslo (Norway), South-East Queensland (Australia) and Eindhoven (the Netherlands)., Measurements: Wastewater data were modelled for MDMA (3,4-methyl​enedioxy​methamphetamine), benzoylecgonine (a metabolite of cocaine), amphetamine and methamphetamine in Oslo; benzoylecgonine in Eindhoven; and methamphetamine in South-East Queensland. Choice of stimulants modelled in each region was primarily determined by availability of useable data., Findings: In Oslo, wastewater data, driving under the influence of drugs statistics and seizure data all suggested increasing MDMA use between 2009 and 2017. In South-East Queensland, there was an estimated 31.1% [95% confidence interval (CI) = 29.4-32.9%] annual increase in daily loads of methamphetamine in wastewater between 2009 and 2016, compared with a 14.1% (95% CI = 10.9-17.3%) annual increase in seizures. Some of the increase in wastewater can be explained by increased purity. In Eindhoven, there was no evidence of a change in cocaine consumption from wastewater, but a reduction was observed in numbers in treatment for cocaine use from 2012 to 2017. In approximately half the cases examined in Oslo, credible intervals around estimates of annual average loads from a regression model versus estimates based on a single week of sampling did not overlap., Conclusions: Long-term trends in loads of stimulants in wastewater appear to be broadly consistent with trends in other indicators of stimulant use in three locations. Wastewater data should be interpreted alongside epidemiological indicators and purity data. One week of wastewater sampling may not be sufficient for valid inference about drug consumption., (© 2019 Society for the Study of Addiction.)

Wastewater-based epidemiology is a growing research field which provides valuable information on community drug use and chemical exposure. One parameter critical to estimations of drug use is the catchment area population. A population biomarker could be used to provide this information. This study evaluated the analytical suitability of three endogenous biomarkers of human activity: the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) which has previously been proposed and two further candidates, the catecholamine metabolites vanillylmandelic acid (VMA) and homovanillic acid (HVA). An analytical method involving derivatization was developed and validated for two candidates, 5-HIAA and HVA by liquid chromatography - mass spectrometry. The best performance was obtained for VMA as the underivatized analyte. The derivatized extracts produced a 100 times better sensitivity. The three neurotransmitter metabolites were evaluated as population biomarkers in wastewater samples. All were stable in sample, not lost upon filtration and showed stable inter-day mass loads over seven days for a metropolitan wastewater treatment plant. When applied to a small community during a festival period, mass loads of both HVA and VMA reflected the increase in the catchment population, whilst 5-HIAA proved to be more variable., Competing Interests: Declaration of Competing Interest All authors have participated in (a) conception and design, or analysis and interpretation of the data; (b) drafting the article or revising it critically for important intellectual content; and (c) approval of the final version. This manuscript has not been submitted to, nor is under review at, another journal or other publishing venue. The authors have no affiliation with any organization with a direct or indirect financial interest in the subject matter discussed in the manuscript, (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Wastewater-based epidemiology (WBE) provides complementary information to traditional self-report methods for estimating substance use within a population. WBE was applied to estimate the consumption of alcohol in an Australian rural city (population estimated 100,000) over 6 years., Methods: A total of 352 wastewater samples were analysed from a wastewater treatment plant located in South-East Queensland, Australia, from 2012 to 2017. The concentration of an alcohol biomarker, ethyl sulphate, was quantified by liquid chromatography tandem mass spectrometry and used to estimate per-capita consumption. The WBE results were compared with alcohol consumption estimates based on national taxation data and self-reported national survey data in Australia., Results: Average daily alcohol consumption estimated by WBE was between 19 and 30 mL/person/day for the population aged 15 years and older during the six-year period. Alcohol consumption decreased 4 % per annum on average over the study period. Our data showed higher rates of consumption on weekends and public holidays when compared to consumption between Monday and Thursday. The comparative trend of WBE data was consistent with the national alcohol survey and taxation statistics on alcoholic beverages over the same period., Conclusions: A clear decline in alcohol consumption in the catchment was observed during the sampling period, which reflected similar changes in consumption from taxation statistics and self-report survey data. Expected variations in weekly consumption and public holidays were also identified. This study demonstrates the potential of WBE for long-term monitoring of alcohol consumption in evaluating the effectiveness of local and national alcohol policies and prevention programs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background and Aims: Wastewater-based epidemiology is an additional indicator of drug use that is gaining reliability to complement the current established panel of indicators. The aims of this study were to: (i) assess spatial and temporal trends of population-normalized mass loads of benzoylecgonine, amphetamine, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) in raw wastewater over 7 years (2011-17); (ii) address overall drug use by estimating the average number of combined doses consumed per day in each city; and (iii) compare these with existing prevalence and seizure data., Design: Analysis of daily raw wastewater composite samples collected over 1 week per year from 2011 to 2017., Setting and Participants: Catchment areas of 143 wastewater treatment plants in 120 cities in 37 countries., Measurements: Parent substances (amphetamine, methamphetamine and MDMA) and the metabolites of cocaine (benzoylecgonine) and of Δ 9 -tetrahydrocannabinol (11-nor-9-carboxy-Δ 9 -tetrahydrocannabinol) were measured in wastewater using liquid chromatography-tandem mass spectrometry. Daily mass loads (mg/day) were normalized to catchment population (mg/1000 people/day) and converted to the number of combined doses consumed per day. Spatial differences were assessed world-wide, and temporal trends were discerned at European level by comparing 2011-13 drug loads versus 2014-17 loads., Findings: Benzoylecgonine was the stimulant metabolite detected at higher loads in southern and western Europe, and amphetamine, MDMA and methamphetamine in East and North-Central Europe. In other continents, methamphetamine showed the highest levels in the United States and Australia and benzoylecgonine in South America. During the reporting period, benzoylecgonine loads increased in general across Europe, amphetamine and methamphetamine levels fluctuated and MDMA underwent an intermittent upsurge., Conclusions: The analysis of wastewater to quantify drug loads provides near real-time drug use estimates that globally correspond to prevalence and seizure data., (© 2019 The Authors. Addiction published by John Wiley & Sons Ltd on behalf of Society for the Study of Addiction.)

Wastewater contains a wealth of information about the population who contribute to it including biological and chemical markers of human activity and exposures. F 2 -isoprostanes have been proposed as oxidative stress biomarkers that can be measured in wastewater to provide a measure of oxidative stress at the population level. While an association between tobacco use and their level in wastewater has been demonstrated, an in-sewer stability assessment has not been conducted to support their use as oxidative stress biomarkers for wastewater-based epidemiology studies. In this study we investigated the stability of 8-iso-prostaglandin F 2α (PGF 2α ), its metabolite dinor-11β-Prostaglandin F 2α (dnPGF 2α ) and Prostaglandin E 2 (PGE 2 ) (representative of other classes of prostaglandins) in laboratory-scale sewer reactors simulating real sewers. PGF 2α , dnPGF 2α and PGE 2 were all found to be sufficiently stable under typical sewer conditions therefore satisfying the stability requirement of wastewater-based epidemiology population health biomarkers., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Wastewater studies that provide per capita estimates of consumption (influent) or release (effluent) via wastewater systems rely heavily on accurate population data. This study evaluated the accuracy of Wastewater Treatment Plant (WWTP) reported populations, as well as hydrochemical parameters, against accurate populations from a population census. 104 catchment maps were received from WWTPs, geolocated in geospatial software and overlaid with the smallest area unit of the Australian census, equating to 14.9 million Australians or 64% of the national population. We characterized each catchment for population counts, as well as by age profile, income profile, and education level. For a subset of sites, population estimates using hydrochemical parameters BOD, COD, and dissolved ammonia were evaluated for accuracy against census populations. Population estimates provided by WWTP personnel were on average 18% higher than census-based populations. Furthermore, hydrochemical-based population estimates had high RSD (>44%) for BOD, COD, and ammonium between sites, suggesting that their applicability for use in population estimation may not be appropriate for every WWTP. Catchment age distributions were evaluated and 46% of catchments had skewed age distributions: 6% were skewed older, and 40% were skewed younger. Through this process WWTP catchment populations can be characterized in a way that will enhance the interpretations of per capita estimates.

Anabolic-androgenic steroids are synthetic compounds prohibited due to their performance-enhancing characteristics. The use of these substances is known to cause health-related issues, which highlights the importance of being able to evaluate the scale of consumption by the general population. However, most available research on the analysis of anabolic steroids is focused on animals and athletes in connection with doping. The potential of wastewater-based epidemiology as an intelligence tool for the assessment of community level use of anabolic steroids is presented herein. A liquid chromatography tandem mass spectrometry method was developed for the analysis of 10 anabolic-androgenic steroids and 14 endogenous hormones in influent wastewater. The validated method was applied to sixteen 24-hour composite wastewater influent samples that were collected over a period of five years from two wastewater treatment plants in Queensland, Australia. Nine investigated compounds were found to be present at concentrations between 14 and 611 ng L -1 which translated into 3-104 mg excreted per 1000 individuals per day. It was concluded that the developed analytical method is suitable for the analysis of AAS in wastewater matrix. Additionally, both the inclusion of metabolites and further investigation into deconjugation by enzymatic hydrolysis would aid in understanding and evaluating community anabolic steroid use. For the first time, this study presents the application of wastewater-based epidemiology on anabolic-androgenic steroids in Australia., (© 2019 John Wiley & Sons, Ltd.)