High throughput and sensitive quantitation of tobacco-specific alkaloids and nitrosamines in wastewater.

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 (R2 > 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.

Analysis of wastewater from 2013 to 2021 detected a recent increase in nicotine use in Queensland, Australia.

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.

Wastewater-based epidemiology of Campylobacter spp.: A systematic review and meta-analysis of influent, effluent, and removal of wastewater treatment plants.

Campylobacter spp. is one of the four leading causes of diarrhoeal diseases worldwide, which are generally mild but can be fatal in children, the elderly, and immunosuppressed persons. The existing disease surveillance for Campylobacter infections is usually based on untimely clinical reports. Wastewater surveillance or wastewater-based epidemiology (WBE) has been developed for the early warning of disease outbreaks and the detection of the emerging new variants of human pathogens, especially after the global pandemic of COVID-19. However, the WBE monitoring of Campylobacter infections in communities is rare due to a few large data gaps. This study is a meta-analysis and systematic review of the prevalence of Campylobacter spp. in various wastewater samples, primarily the influent of wastewater treatment plants. The results showed that the overall prevalence of Campylobacter spp. was 53.26 % in influent wastewater and 52.97 % in all types of wastewater samples. The mean concentration in the influent was 3.31 ± 0.39 log10 gene copies or most probable number (MPN) per 100 mL. The detection method combining culture and PCR yielded the highest positive rate of 90.86 %, while RT-qPCR and qPCR were the two most frequently used quantification methods. In addition, the Campylobacter concentration in influent wastewater showed a seasonal fluctuation, with the highest concentration in the autumn at 3.46 ± 0.41 log10 gene copies or MPN per 100 mL. Based on the isolates of all positive samples, Campylobacter jejuni (62.34 %) was identified as the most prevalent species in wastewater, followed by Campylobacter coli (30.85 %) and Campylobacter lari (4.4 %). These findings provided significant data to further develop and optimize the wastewater surveillance of Campylobacter spp. infections. In addition, large data gaps were found in the decay of Campylobacter spp. in wastewater, indicating insufficient research on the persistence of Campylobacter spp. in wastewater.

There's something in the air: A review of sources, prevalence and behaviour of microplastics in the atmosphere.

Literature regarding microplastics in the atmosphere has advanced in recent years. However, studies have been undertaken in isolation with minimal collaboration and exploration of the relationships between air, deposition and dust. This review collates concentrations (particle count and mass-based), shape, size and polymetric characteristics for microplastics in ambient air (m3), deposition (m2/day), dust (microplastics/g) and snow (microplastics/L) from 124 peer-reviewed articles to provide a holistic overview and analysis of our current knowledge. In summary, ambient air featured concentrations between <1 to >1000 microplastics/m3 (outdoor) and <1 microplastic/m3 to 1583 ± 1181 (mean) microplastics/m3 (indoor), consisting of polyethylene terephthalate, polyethylene, polypropylene. No difference (p > 0.05) was observed between indoor and outdoor concentrations or the minimum size of microplastics (p > 0.5). Maximum microplastic sizes were larger indoors (p < 0.05). Deposition concentrations ranged between 0.5 and 1357 microplastics/m2/day (outdoor) and 475 to 19,600 microplastics/m2/day (indoor), including polyethylene, polystyrene, polypropylene, polyethylene terephthalate. Concentrations varied between indoor and outdoor deposition (p < 0.05), being more abundant indoors, potentially closer to sources/sinks. No difference was observed between the minimum or maximum reported microplastic sizes within indoor and outdoor deposition (p > 0.05). Road dust concentrations varied between 2 ± 2 and 477 microplastics/g (mean), consisting of polyvinyl chloride, polyethylene, polypropylene. Mean outdoor dust concentrations ranged from <1 microplastic/g (remote desert) to between 18 and 225 microplastics/g, comprised of polyethylene terephthalate, polyamide, polypropylene. Snow concentrations varied between 0.1 and 30,000 microplastics/L, containing polyethylene, polyamide, polypropylene. Concentrations within indoor dust varied between 10 and 67,000 microplastics/g, including polyethylene terephthalate, polyethylene, polypropylene. No difference was observed between indoor and outdoor concentrations (microplastics/g) or maximum size (p > 0.05). The minimum size of microplastics were smaller within outdoor dust (p > 0.05). Although comparability is hindered by differing sampling methods, analytical techniques, polymers investigated, spectral libraries and inconsistent terminology, this review provides a synopsis of knowledge to date regarding atmospheric microplastics.

Socioeconomic status and public health in Australia: A wastewater-based study.

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.

Does size matter? Quantification of plastics associated with size fractionated biosolids.

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 (Σ7plastics), 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.

Plastics contamination of store-bought rice.

This study investigated mass concentrations of selected plastics in store-bought rice, the staple of more than half the world's population. Polyethylene, polyethylene terephthalate, poly-(methyl methacrylate), polypropylene, polystyrene and polyvinyl chloride were quantified using pressurized liquid extraction coupled to double-shot pyrolysis gas chromatography/mass spectrometry. Polyethylene, polypropylene and polyethylene terephthalate were quantifiable in the rice samples with polyethylene the most frequently detected (95%). There was no statistical difference between total plastic concentration in paper and plastic packaged rice. Shaking the rice in its packaging had no significant difference on the concentration of plastics. Washing the rice with water significantly reduced plastic contamination. Instant (pre-cooked) rice contained fourfold higher levels of plastics, suggesting that industrial processing potentially increases contamination. A preliminary estimate of the intake of plastic through rice consumption for Australians established 3.7 mg per serve (100 g) if not washed and 2.8 mg if washed. Annual consumption was estimated around 1 g/person.

Quantification of selected microplastics in Australian urban road dust.

Microplastics (1 - 5000 µm) are pervasive in every compartment of our environment. However, little is understood regarding the concentration and size distribution of microplastics in road dust, and how they change in relation to human activity. Within road dust, microplastics move through the environment via atmospheric transportation and stormwater run-off into waterways. Human exposure pathways to road dust include dermal contact, inhalation and ingestion. In this study, road dust along an urban to rural transect within South-East Queensland, Australia was analysed using Accelerated Solvent Extraction followed by pyrolysis Gas Chromatography-Mass Spectrometry (Pyr-GC/MS). Polypropylene, polystyrene, polyethylene terephthalate, polyvinyl chloride, poly (methyl methacrylate) and polyethylene were quantified. Microplastic concentrations ranged from ~0.5 mg/g (rural site) to 6 mg/g (Brisbane city), consisting primarily of polyvinyl chloride (29%) and polyethylene terephthalate (29%). Size fractionation (< 250 µm, 250-500 µm, 500-1000 µm, 1000-2000 µm and 2000-5000 µm) established that the < 250 µm size fraction contained the majority of microplastics by mass (mg/g). Microplastic concentrations in road dust demonstrated a significant relationship with the volume of vehicles (r2 = 0.63), suggesting traffic, as a proxy for human movement, is associated with increased microplastic concentrations in the built environment.

Application of catecholamine metabolites as endogenous population biomarkers for wastewater-based epidemiology.

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 (r2) 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.

SARS-CoV-2 RNA monitoring in wastewater as a potential early warning system for COVID-19 transmission in the community: A temporal case study.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus which causes coronavirus disease (COVID-19), has spread rapidly across the globe infecting millions of people and causing significant health and economic impacts. Authorities are exploring complimentary approaches to monitor this infectious disease at the community level. Wastewater-based epidemiology (WBE) approaches to detect SARS-CoV-2 RNA in municipal wastewater are being implemented worldwide as an environmental surveillance approach to inform health authority decision-making. Owing to the extended excretion of SARS-CoV-2 RNA in stool, WBE can surveil large populated areas with a longer detection window providing unique information on the presence of pre-symptomatic and asymptomatic cases that are unlikely to be screened by clinical testing. Herein, we analysed SARS-CoV-2 RNA in 24-h composite wastewater samples (n = 63) from three wastewater treatment plants (WWTPs) in Brisbane, Queensland, Australia from 24th of February to 1st of May 2020. A total of 21 samples were positive for SARS-CoV-2, ranging from 135 to 11,992 gene copies (GC)/100 mL of wastewater. Detections were made in a Southern Brisbane WWTP in late February 2020, up to three weeks before the first clininal case was reported there. Wastewater samples were generally positive during the period with highest caseload data. The positive SARS-CoV-2 RNA detection in wastewater while there were limited clinical reported cases demonstrates the potential of WBE as an early warning system to identify hotspots and target localised public health responses, such as increased individual testing and the provision of health warnings.

Long-term trends in tobacco use assessed by wastewater-based epidemiology and its relationship with consumption of nicotine containing products.

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.

Quantitative Analysis of Selected Plastics in High-Commercial-Value Australian Seafood by Pyrolysis Gas Chromatography Mass Spectrometry.

Microplastic contamination of the marine environment is widespread, but the extent to which the marine food web is contaminated is not yet known. The aims of this study were to go beyond visual identification techniques and develop and apply a simple seafood sample cleanup, extraction, and quantitative analysis method using pyrolysis gas chromatography mass spectrometry to improve the detection of plastic contamination. This method allows the identification and quantification of polystyrene, polyethylene, polyvinyl chloride, polypropylene, and poly(methyl methacrylate) in the edible portion of five different seafood organisms: oysters, prawns, squid, crabs, and sardines. Polyvinyl chloride was detected in all samples and polyethylene at the highest total concentration of between 0.04 and 2.4 mg g-1 of tissue. Sardines contained the highest total plastic mass concentration (0.3 mg g-1 tissue) and squid the lowest (0.04 mg g-1 tissue). Our findings show that the total concentration of plastics is highly variable among species and that microplastic concentration differs between organisms of the same species. The sources of microplastic exposure, such as packaging and handling with consequent transference and adherence to the tissues, are discussed. This method is a major development in the standardization of plastic quantification techniques used in seafood.

Determination of anabasine, anatabine, and nicotine biomarkers in wastewater by enhanced direct injection LC-MS/MS and evaluation of their in-sewer stability.

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.

Identification and quantification of selected plastics in biosolids by pressurized liquid extraction combined with double-shot pyrolysis gas chromatography-mass spectrometry.

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 Æ©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.

Evaluating the stability of three oxidative stress biomarkers under sewer conditions and potential impact for use in wastewater-based epidemiology.

Wastewater contains a wealth of information about the population who contribute to it including biological and chemical markers of human activity and exposures. F2-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 F2α (PGF2α), its metabolite dinor-11ß-Prostaglandin F2α (dnPGF2α) and Prostaglandin E2 (PGE2) (representative of other classes of prostaglandins) in laboratory-scale sewer reactors simulating real sewers. PGF2α, dnPGF2α and PGE2 were all found to be sufficiently stable under typical sewer conditions therefore satisfying the stability requirement of wastewater-based epidemiology population health biomarkers.

Chlorinated paraffins in indoor dust from Australia: Levels, congener patterns and preliminary assessment of human exposure.

Chlorinated paraffins (CPs) are a group of polychlorinated n-alkanes with high production volumes. Until now, there are only limited data on the levels of CPs in the environment, especially in the indoor environment. In this study, dust samples were collected from 44 indoor environments, including 27 private houses, 10 offices, and 7 vehicles. Short-, medium-, and long-chain CPs were detected in all dust samples. The median concentration of ∑CPs (C10-C21) was 57, 160 and 290 µg/g, in houses, offices, and vehicles, respectively. Medium-chain CPs were the dominant group, on average accounting for 86% of ∑CPs. Cl6 and Cl8 groups had the highest contributions to ∑CPs across all the different microenvironments, while C13 and C14 were the predominant groups of SCCPs and MCCPs. Median exposure to ∑CPs via indoor dust were estimated at 80 ng/kg/day and 620 µg/kg/day for Australian adults and toddlers respectively. The daily intake of CPs via dust, in the worse scenario, was still 2-3 orders of magnitudes lower than the reference doses based on neoplastic effects.

Trends in nicotine consumption between 2010 and 2017 in an Australian city using the wastewater-based epidemiology approach.

Monitoring smoking prevalence is key to assessing responses to tobacco control measures, and evaluating associated health and economic costs. Estimates of tobacco consumed in Australia are based on various data sources - tax excise clearances, sales, and self-report surveys. There are limitations with each of these data sources which makes triangulation of cigarette use estimates by multiple methods important. Wastewater-based epidemiology, the systematic sampling and analysis of wastewater, is now a routine method to measure and monitor human exposure to a range of chemicals. This study provides a high frequency long-term temporal assessment of exposure to nicotine, the main addictive component of tobacco, using this approach. 291 archived wastewater samples collected from a regional city catchment from 2010 to 2017 were analysed for human-specific nicotine metabolites (cotinine and trans-3'-hydroxycotinine), to estimate per capita nicotine use. Temporal trends in nicotine use determined by wastewater-based epidemiology were compared with national sales and survey data. Wastewater analysis showed a 25% reduction in the mean number of cigarette equivalents consumed from 2010 to 2017, representing a 3% annual decline. These findings are in good agreement with estimates based on surveys and sales data, indicating annual declines of 5% and 4%, respectively. Findings of this study demonstrate WBE to be a relatively cost-effective and objective approach to reporting long-term data on nicotine consumption. When combined with alternative data sources, and valuable sociodemographic information of surveys, wastewater-based epidemiology helps to refine our estimates and understanding of the total impacts of smoking.

Population histamine burden assessed using wastewater-based epidemiology: The association of 1,4­methylimidazole acetic acid and fexofenadine.

Systematic sampling and analysis of wastewater has become an important tool for monitoring consumption of drugs and other substances, and has been proposed as a method to evaluate aspects of population health using endogenous biomarkers. 1,4­methylimidazoleacetic acid (MIAA) is an endogenous biomarker and metabolite of histamine turnover. Its urinary excretion is elevated in conditions such as mastocytosis, hay fever, hives, food allergies and anaphylaxis. The aim of this study was to develop and apply methods for MIAA in wastewater and compare its occurrence with antihistamine use in wastewater. Consecutive daily samples were collected from seven catchments serving populations from 3000 to 2 million and covering rural and urban communities during the 2016 Census in Australia. MIAA and the antihistamines (ranitidine, fexofenadine, cetirizine) were quantified consistently. Per capita excretion of MIAA (mg/d/capita) estimated from the WW concentrations were consistent with findings from previous clinical studies. We found significant positive correlations between loads of MIAA and fexofenadine (R2 = 0.68, p < 0.0001) and cetirizine (R2 = 0.25, p = 0.03) across the various catchments. Sewer reactor experiments on the degradation of MIAA and the antihistamines found that fexofenadine is stable for at least 24 h while MIAA, ranitidine and cetirizine are subject to degradation, and this should be considered in interpretations. To the best of our knowledge, this study is the first wastewater study to introduce and monitor an endogenous metabolite of histamine, and the first study to monitor and relate proxies of disease and treatment of disease.