Characterising differences between self-reported and wastewater-identified drug use at two consecutive years of an Australian music festival.

BackgroundIn the context of drug prohibition, potential adulteration and variable purity pose additional health risks for people who use drugs, with these risks often compounded by the outdoor music festival environment. Ahead of the imminent implementation of drug checking services in Queensland, Australia, this study aims to characterise this problem using triangulated survey and wastewater data to understand self-reported and detected drug use among attendees of a multi-day Queensland-based music festival in 2021 and 2022.MethodsWe administered an in-situ survey focusing on drug use at the festival to two convenience samples of 136 and 140 festival attendees in 2021 and 2022 respectively. We compared survey findings to wastewater collected concurrently from the festival's site-specific wastewater treatment plant, which was analysed using Liquid Chromatography Tandem Mass Spectrometry.ResultsMost survey respondents (82 % in 2021, 92 % in 2022) reported using or intending to use an illicit drug at the festival. Some respondents reported potentially risky drug use practices such as using drugs found on the ground (2 % in 2021, 4 % in 2022). Substances detected in wastewater but not surveys include MDEA, mephedrone, methylone, 3-MMC, alpha-D2PV, etizolam, eutylone, and N,N-dimethylpentylone.ConclusionMany substances detected in wastewater but not self-reported in surveys likely represent substitutions or adulterants. These findings highlight the benefits of drug checking services to prevent harms from adulterants and provide education on safer drug use practices. These findings also provide useful information on socio-demographic characteristics and drug use patterns of potential users of Queensland's future drug checking service

Annual release of selected UV filters via effluent from wastewater treatment plants in Australia

Studies conducted globally have identified wastewater effluent as a key source of UV filters released into the aquatic environment. We assessed the annual release of UV filters from wastewater treatment plant effluent in Australia and evaluated the removal of these chemicals during wastewater treatment. Effluent samples were collected from 33 sites alongside matching influent samples. Sample collection predominately occurred during the Australian Census in August 2016, which allowed for accurate per capita normalisation of the results. A subset of sites was also sampled over the Southern Hemisphere summer (December-February) period. Five UV filters were detected with at least one detected in 95% of effluent samples. The summed concentration of UV filters ranged from 130\ua0ng\ua0L to 8400\ua0ng\ua0L and averaged 2800 (±1900) ng L. Of the target UV filters, 2-phenylbenzimidazole-5-sulfonic acid (PBSA) and benzophenone 4 (BP4) showed the lowest removal efficiencies (11\ua0±\ua036% and 51\ua0±\ua043%, respectively) across all sites and were the most abundant in effluent. Average estimated removal efficiencies of the other compounds were between 59 (±24) % (4-methylbenzylidene camphor (4-MBC)) and 74 (±22) % (benzophenone 1 (BP1)). We did not find a trend in seasonal differences in the per capita release of UV filters in effluent samples. We estimate that approximately 40% of UV filter loads measured in influent are breaking through to the effluent resulting in the release of approximately 20\ua0kg day of the selected UV filters into the aquatic environment from treated wastewater effluent in Australia

Calibration parameters for the passive sampling of organic UV filters by silicone; diffusion coefficients and silicone–water partition coefficients

In recent years, organic ultraviolet filters (UVFs) received considerable attention as a group of emerging contaminants, including in Australia where the use of UVFs is particularly relevant. Passive sampling using polymers has become widely used for routine monitoring of chemicals in the aquatic environment. Application of passive samplers for monitoring chemicals in the water relies on calibration data such as chemical's polymer-water partition coefficient (K) and diffusion coefficients in the sampling material (D), for understanding uptake and kinetic limitations. In the present study, K and D for nine UVFs were estimated. K values were determined in different water - polymer partition experiments where (1) a given mass of chemicals was dosed into the water and (2) into the polymer. Diffusion coefficients were determined using the stacking method. The estimated log K and log D ranged from 2.9 to 6.4 L kg and -11.1 to\ua0-10.5 ms, respectively. The sufficient high D allows application of kinetic models that only consider water boundary-controlled uptake for converting silicone sampler uptake into an aqueous phase concentration using the presented K

The presence of selected UV filters in a freshwater recreational reservoir and fate in controlled experiments

UV filters present in sunscreen and other cosmetics are directly released into the environment during aquatic recreational activities. The extent to which the wide range of UV filters pose a risk to the environment remains unclear. This study investigated the occurrence and dissipation of selected organic UV filters at a recreational site (Enoggera Reservoir, Queensland, Australia) over 12 h. Furthermore, different possible degradation processes were investigated in a controlled off-site experiment with surface water exposed to natural light. Half-lives were estimated for ten UV filters. In Enoggera Reservoir, seven UV filters were detected, of which the most prevalent were octocrylene, avobenzone (BMDBM) and enzacamene (4-MBC). Summed concentrations of the seven UV filters ranged from 7330 ng L at 13:00 h to 2550 ng L at 21:00 h. In the degradation experiment, four UV filters showed no significant change over time. The fate of these compounds in the environment is likely to be mainly influenced by dispersion. Half-lives of the remaining UV filters were 6.6 h for amiloxate (IMC), 20 h for benzophenone 1, 23 h for octinoxate (EHMC), 30 h for 3-benzylidene camphor, 34 h for 4-MBC and 140 h for dioxybenzone (BP8). The degree of susceptibility to photodegradation and biodegradation was generally consistent within a structural class. The fate and half-lives of UV filters are variable and should be considered on a per site basis when assessing environmental risk

Improving Wastewater-Based Epidemiology for New Psychoactive Substance Surveillance by Combining a High-Throughput In Vitro Metabolism Assay and LC−HRMS Metabolite Identification

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

In Situ Calibration of Passive Samplers for Viruses in Wastewater

Wastewater surveillance for SARS-CoV-2 RNAhas rapidly developed worldwide. In low-prevalence settings, sampling in sewage networks is proposed to monitor community transmission. Passive samplers are cost-effective and suitable for catchments where autosamplers cannot be operated. This resulted in their pioneering applications in some countries, even though their sampling kinetics for viruses remains unclear. We conducted in situ calibration of passive sampling materials (membranes, swabs, gauzes, and tampons) for the uptake of pepper mild mottle virus (PMMoV), enterovirus, and human adenovirus 40/41. Passive samplers were deployed in wastewater influent and retrieved sequentially over 48 h. Membranes performed continuous sampling over 48 h with estimated linear sampling rates of 1 mL h-1for PMMoV, 0.3 mL h-1for enterovirus, and 33.1 mL h-1for adenovirus. Tampons and swabs showed a rapid initial uptake of viruses and reached equilibrium after 8 h, while gauze uptake rates were potentially confounded by either inhibitors or viral losses during extended exposure. Additionally, monitoring SARS-CoV-2 at 17 sewer manholes showed that the detection ratio of membranes (14 of 17) was higher than that of tampons (8 of 17). This study demonstrated the ability of passive samplers to retain viral fragments, making them a practical tool for wastewater surveillance for the detection of disease outbreaks in communities.</p

Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater

There is currently a clear benefit for many countries to utilize wastewater-based epidemiology (WBE) as part of ongoing measures to manage the coronavirus disease 2019 (COVID-19) global pandemic. Since most wastewater virus concentration methods were developed and validated for nonenveloped viruses, it is imperative to determine the efficiency of the most commonly used methods for the enveloped severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Municipal wastewater seeded with a human coronavirus (CoV) surrogate, murine hepatitis virus (MHV), was used to test the efficiency of seven wastewater virus concentration methods: (A–C) adsorption-extraction with three different pre-treatment options, (D–E) centrifugal filter device methods with two different devices, (F) polyethylene glycol (PEG 8000) precipitation, and (G) ultracentrifugation. MHV was quantified by reverse-transcription quantitative polymerase chain reaction and the recovery efficiency was calculated for each method. The mean MHV recoveries ranged from 26.7 to 65.7%. The most efficient methods were adsorption-extraction methods with MgCl2 pre-treatment (Method C), and without pre-treatment (Method B). The third most efficient method used the Amicon® Ultra-15 centrifugal filter device (Method D) and its recovery efficiency was not statistically different from the most efficient methods. The methods with the worst recovery efficiency included the adsorption-extraction method with acidification (A), followed by PEG precipitation (F). Our results suggest that absorption-extraction methods with minimal or without pre-treatment can provide suitably rapid, cost-effective and relatively straightforward recovery of enveloped viruses in wastewater. The MHV is a promising process control for SARS-CoV-2 surveillance and can be used as a quality control measure to support community-level epidemic mitigation and risk assessment

First confirmed detection of SARS-CoV-2 in untreated wastewater in Australia: A proof of concept for the wastewater surveillance of COVID-19 in the community

Infection with SARS-CoV-2, the etiologic agent of the ongoing COVID-19 pandemic, is accompanied by the shedding of the virus in stool. Therefore, the quantification of SARS-CoV-2 in wastewater affords the ability to monitor the prevalence of infections among the population via wastewater-based epidemiology (WBE). In the current work, SARS-CoV-2 RNA was concentrated from wastewater in a catchment in Australia and viral RNA copies were enumerated using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) resulting in two positive detections within a six day period from the same wastewater treatment plant (WWTP). The estimated viral RNA copy numbers observed in the wastewater were then used to estimate the number of infected individuals in the catchment via Monte Carlo simulation. Given the uncertainty and variation in the input parameters, the model estimated a median range of 171 to 1,090 infected persons in the catchment, which is in reasonable agreement with clinical observations. This work highlights the viability of WBE for monitoring infectious diseases, such as COVID-19, in communities. The work also draws attention to the need for further methodological and molecular assay validation for enveloped viruses in wastewater