Influent Samples Research Articles

Potential Exposure to Respiratory and Enteric Bacterial Pathogens among Wastewater Treatment Plant Workers, South Africa

Wastewater handling has been associated with an increased risk of developing adverse health effects, including respiratory and gastrointestinal illnesses. However, there is a paucity of information in the literature, and occupational health risks are not well quantified. Grab influent samples were analysed using Illumina Miseq 16S amplicon sequencing to assess potential worker exposure to bacterial pathogens occurring in five municipal wastewater treatment plants (WWTPs). The most predominant phyla were Bacteroidota, Campilobacterota, Proteobacteria, Firmicutes, and Desulfobacterota, accounting for 85.4% of the total bacterial community. Taxonomic analysis showed a relatively low diversity of bacterial composition of the predominant genera across all WWTPs, indicating a high degree of bacterial community stability in the influent source. Pathogenic bacterial genera of human health concern included Mycobacterium, Coxiella, Escherichia/Shigella, Arcobacter, Acinetobacter, Streptococcus, Treponema, and Aeromonas. Furthermore, WHO-listed inherently resistant opportunistic bacterial genera were identified. These results suggest that WWTP workers may be occupationally exposed to several bacterial genera classified as hazardous biological agents for humans. Therefore, there is a need for comprehensive risk assessments to ascertain the actual risks and health outcomes among WWTP workers and inform effective intervention strategies to reduce worker exposure.

Application of PARAFAC Technique to Compare Fluorescence Property of Different Dissolved Organic Matters

Objectives : It provides a detailed procedure for analyzing dissolved organic matter (DOM) by the PARAFAC technique and aims to identify compare different fluorescent components of DOM with various water sources.Methods : PARAFAC modeling was performed based on the PARAFAC tutorial by Stedmon et al. [11] using a total of 160 samples originated from wetlands, river, wastewater treatment plant influent and effluent. The procedure includes data preprocessing, initial model creation, outlier analysis, split-half analysis, core consistency, and sum of squared error analysis (SSE). After finishing the model validation step, the samples were analyzed through the verified PARAFAC model.Results and Discussion : An initial model with 2 to 4 components was created, and as a result of outlier analysis, the four WWTP Influent samples were removed as outliers. As a result of SSE analysis, the model with two components has a relatively higher SSE value than the model with three or four components. In the case of core consistency, the model with 2, 3, and 4 components scored 99.6%, 94.9%, and 65.7%, respectively. As a result of the split-half analysis, the spectra of the model having the three components of the split data were the same, so it was finally determined that the number of three components was appropriate in the model. The DOM samples were analyzed by the developed PARAFAC model. It was confirmed that the ratio of humic components was higher in the order of WWTP effluent ≈ wetland > river > WWTP influent, and the ratio of protein components affected by microbial activity was in the order of WWTP influent > river > wetland ≈ WWTP effluent.Conclusion : In this paper, the procedure of PARAFAC modeling was described and the DOM fluorescence properties were compared. This research provides a practical method for increasing the utilization of the PARAFAC model in the water treatment research field. In addition, the result of the PARAFAC model is useful data to control DOM efficiently in water treatment facilities.

Do the lockdown-imposed changes in a wastewater treatment plant catchment's socio-demographics impact longitudinal temporal trends in psychoactive pharmaceutical use?

Wastewater-based epidemiology (WBE) includes the analysis of human metabolic biomarkers of xenobiotics in influent wastewater. WBE complements existing drug utilization approaches and provides objective, spatio-temporal information on the consumption of pharmaceuticals in the general population. This approach was applied to 24-h composite influent wastewater samples from Leuven, Belgium. Daily samples were analysed from September 2019 to December 2019 (n = 76), and on three days of the week (Monday, Wednesday, Saturday) from January 2020 to April 2022 (n = 367). Sample analysis consisted of 96-well solid-phase extraction and liquid chromatography coupled to tandem mass spectrometry. Measured concentrations of 21 biomarkers for antidepressant and opioid use were converted to population-normalized mass loads (PNML) by considering the flow rate and catchment population. To capture population movements, mobile phone data was used. Amitriptyline, hydroxy-bupropion, norcitalopram, citalopram, normirtazapine, trazodone, O-desmethylvenlafaxine, codeine, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), methadone, morphine, O-desmethyltramadol, and tramadol were included in the temporal assessment since concentrations were above the lower limit of quantification. The PNML of most biomarkers increased (with 3–119 %) throughout the sampling period. The population disruption during the COVID-19 pandemic led to a major change in the socio-demographics of the catchment area, resulting in temporal differences in the PNML of the different biomarkers. As such, higher PNML were observed during the different lockdown phases, which were characterized by the outflow of university students and a decreasing commuting in and out the catchment area. The effects of the fluctuating socio-demographics of the catchment population were further evidenced by the different week-weekend pattern of PNMLs over the course of the sampling campaign. Mean parent/metabolite ratios (i.e., citalopram/norcitalopram, tramadol/O-desmethyltramadol, venlafaxine/O-desmethylvenlafaxine, and methadone/EDDP) remained relatively stable throughout the entire sampling campaign (RSD% below 25 % for all ratios, except for methadone/EDDP) and therefore were not affected by this population change.

Detection of the Omicron BA.1 Variant of SARS-CoV-2 in Wastewater From a Las Vegas Tourist Area

Interpretation of wastewater surveillance data is potentially confounded in communities with mobile populations, so it is important to account for this issue when conducting wastewater-based epidemiology (WBE). To leverage spatial and temporal differences in wastewater whole-genome sequencing (WGS) data to quantify relative SARS-CoV-2 contributions from visitors to southern Nevada. This cross-sectional wastewater surveillance study was performed during the COVID-19 pandemic (March 2020 to February 2022) and included weekly influent wastewater samples that were analyzed by reverse transcription-quantitative polymerase chain reaction to quantify SARS-CoV-2 RNA and WGS for identification of variants of concern. This study was conducted in the Las Vegas, Nevada, metropolitan area, which is a semi-urban area with approximately 2.3 million residents and nearly 1 million weekly visitors. Samples were collected from 7 wastewater treatment plant (WWTP) locations that collectively serve the vast majority of southern Nevada (excluding the small number of septic systems) and 1 manhole serving the southern portion of the Las Vegas Strip. With Las Vegas tourism returning to prepandemic levels in 2021, it was hypothesized that visitors were contributing a disproportionate fraction of SARS-CoV-2 RNA to the largest WWTP in southern Nevada, potentially confounding efforts to estimate COVID-19 incidence in the local community through WBE. Relative SARS-CoV-2 load and variants from visitors vs the local population. The Omicron BA.1 VOC was detected in the Las Vegas Strip manhole approximately 1 week before its detection at the WWTP locations (December 13, 2021) and by clinical testing (December 14, 2021). On December 13, Omicron-specific mutations represented a mean (SD) of 48.0% (4.2%) of all genomes from the Las Vegas Strip manhole and 4.1% (1.4%) of all genomes at facilities 2 and 3; by December 20, Omicron-specific mutations represented means (SD) of 82.0% (3.0%) of all genomes at the Las Vegas Strip manhole and 48.0% (2.8%) of all genomes at facilities 2 and 3, respectively. During this time, it was estimated that visitors contributed more than 60% of the SARS-CoV-2 load to the sewershed serving the Las Vegas Strip and that Omicron prevalence among visitors was 40% to 60% on December 13 and 80% to 100% on December 20th. Wastewater surveillance is a valuable complement to clinical tools and can provide time-sensitive data for decision-makers and policy makers. This study represents a novel approach for quantifying the confounding effects of mobile populations on wastewater surveillance data, thereby allowing for modification of an existing WBE framework for estimating COVID-19 incidence in southern Nevada.

Applicability of crAssphage as a performance indicator for viral reduction during activated sludge wastewater treatment

A major threat to water quality is the discharge of human-derived wastewater, which can cause waterborne illnesses associated with enteric viruses. A poor association exists between fecal indicator bacteria and virus fate in the environment, especially during wastewater treatment. In the current study, the potential of using a novel human gut bacteriophage crAssphage as a wastewater treatment process indicator was evaluated. Using qPCR, influent and effluent wastewater samples of two wastewater treatment plants were analyzed for crAssphage and human viruses including human bocavirus (HBoV), human adenovirus (HAdV), and human polyomavirus (HPyV). All samples were positive for crAssphage. The annual crAssphage concentrations varied between 1.45E + 04 and 2.39E + 08 gc/l in influent samples and from 1.25E + 04 to 7.88E + 06 gc/l in effluent samples. Human viruses concentrations were some orders of magnitude lower than that of crAssphage. Data demonstrated a significant correlation between crAssphage, HAdV, and HPyV during the wastewater treatment process, suggesting that crAssphage and human viral pathogens have similar removal mechanisms. Ultimately, this work concludes that crAssphage could be a performance indicator for viral reduction in the wastewater treatment process.

Heavy metals in influent and effluent from 146 drinking water treatment plants across China: Occurrence, explanatory factors, probabilistic health risk, and removal efficiency

Heavy metals (HMs) in drinking water have drawn worldwide attention due to their risks to public health; however, a systematic assessment of the occurrence of HMs in drinking water treatment plants (DWTPs) at a large geographical scale across China and the removal efficiency, human health risks, and the correlation with environmental factors have yet to be established. Therefore, this study characterised the occurrence patterns of nine conventional dissolved HMs in the influent and effluent water samples from 146 typical DWTPs in seven major river basins across China (which consist of the Yangtze River, the Yellow River, the Songhua River, the Pearl River, the Huaihe River, the Liaohe River and the Haihe River) for the first time and removal efficiency, probabilistic health risks, and the correlation with water quality. According to the findings, a total of eight HMs (beryllium (Be), antimony (Sb), barium (Ba), molybdenum (Mo), nickel (Ni), vanadium (V), cobalt (Co) and titanium (Ti)) were detected, with detection frequencies in influent and effluent water ranging from 2.90 (Mo) to 99.30% (Ba) and 1.40 (Ti) to 97.90% (Ba), respectively. The average concentration range was 0.41 (Be)− 77.36 (Sb) μg/L. Among them, Sb (exceeding standard rate 8%), Ba (2.89%), Ni (21.43%), and V (1.33%) were exceeded the national standard (GB5749–2022). By combining Spearman’s results and redundancy analysis, our results revealed a close correlation among pH, turbidity (TURB), potassium permanganate index (CODMn), and total nitrogen (TN) along with the concentration and composition of HMs. In addition, the concentration of HMs in finished water was strongly affected by the concentration of HMs in raw water, as evidenced by the fact that HMs in surface water poses a risk to the quality of finished water. Metal concentration was the primary factor in assessing the health risk of a single metal, and the carcinogenic risk of Ba, Mo, Ni, and Sb should be paid attention to. In DWTPs, the removal efficiencies of various HMs also vary greatly, with an average removal rate ranging from 16.30% to 95.64%. In summary, our findings provide insights into the water quality and health risks caused by HMs in drinking water.

Surveillance on ESBL-Escherichia coli and Indicator ARG in Wastewater and Reclaimed Water of Four Regions of Spain: Impact of Different Disinfection Treatments

In the present study, the occurrence of indicator antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) both in the influent and the effluent of four Spanish wastewater treatment plants (WWTPs) was monitored for 12 months, and the susceptibility profiles of 89 recovered extended spectrum β-lactamase (ESBL)-producing Escherichia coli isolates were obtained against a wide range of antimicrobials. The aim of the study was to better understand whether the current wastewater treatment practices allow us to obtain safe reclaimed water mitigating the spread of ARB and ARGs to the environment. Results showed high concentrations of ESBL-producing E. coli as well as a high prevalence of a range of ARGs in the influent samples. The reclamation treatments implemented in the WWTPs were effective in reducing both the occurrence of ESBL E. coli and ARGs, although significant differences were observed among WWTPs. Despite these reductions in occurrence observed upon wastewater treatment, our findings suggest that WWTP effluents may represent an important source of ARGs, which could be transferred among environmental bacteria and disseminate antimicrobial resistance through the food chain. Remarkably, no major differences were observed in the susceptibility profiles of the ESBL E. coli isolated from influent and effluent waters, indicating that water treatments do not give rise to the emergence of new resistance phenotypes.

Metagenome sequencing to unveil the occurrence and distribution of antibiotic resistome and in a wastewater treatment plant

ABSTRACT The emergence and persistence of antibiotic resistance genes (ARGs) in wastewater treatment plants (WWTPs) has aroused growing public concern for its risk to human health and ecological safety. Moreover, heavy metals concentrated in sewage and sludge could potentially favour co-selection of ARGs and heavy metal resistance genes (HMRGs). In this study, the profile and abundance of antibiotic and metal resistance genes in influent, sludge and effluent were characterized based on the Structured ARG Datebase (SARG) and Antibacterial Biocide and Metal Resistance Gene Datebase (BacMet) by metagenomic analysis. Sequences were aligning against the INTEGRALL, ISFinder, ICEberg and NCBI RefSeq databases to obtain the diversity and abundance of mobile genetic elements (MGEs, e.g.plasmid and transposon). Among them, 20 types of ARGs and 16 types of HMRG were detected in all samples, the influent metagenomes contained many more resistance genes (both ARGs and HMRGs) than the sludge and the influent sample, large reductions in the relatively abundance and diversity of ARG were achieved by biological treatment. ARGs and HMRGs cannot be completely eliminated during the oxidation ditch. A total of 32 species of the potential pathogens were detected, relative abundances of pathogens had no obvious changes. It is suggested that more specific treatments are required to limit their proliferation in the environment. This study can be helpful for further understanding the removal of antibiotic resistance genes in the sewage treatment process via metagenomic sequencing.

Occurrence of quantifiable and semi-quantifiable poly- and perfluoroalkyl substances in united states wastewater treatment plants

Both quantifiable and semi-quantifiable poly- and perfluoroalkyl substances (PFAS) were evaluated in the influent, effluent, and biosolids of 38 wastewater treatment plants. PFAS were detected in all streams at all facilities. For the means of the sums of detected, quantifiable PFAS concentrations were 98 ± 28 ng/L, 80 ± 24 ng/L, and 160,000 ± 46,000 ng/kg (dry weight basis) in the influent, effluent, and biosolids (respectively). In the aqueous influent and effluent streams this quantifiable PFAS mass was typically associated with perfluoroalkyl acids (PFAAs). In contrast, quantifiable PFAS in the biosolids were primarily polyfluoroalkyl substances that potentially serve as precursors to the more recalcitrant PFAAs. Results of the total oxidizable precursor (TOP) assay on select influent and effluent samples showed that semi-quantified (or, unidentified) precursors accounted for a substantial portion (21 to 88%) of the fluorine mass compared to that associated with quantified PFAS, and that this fluorine precursor mass was not appreciably transformed to perfluoroalkyl acids within the WWTPs, as influent and effluent precursor concentrations via the TOP assay were statistically identical. Evaluation of semi-quantified PFAS, consistent with results of the TOP assay, showed the presence of several classes of precursors in the influent, effluent, and biosolids; perfluorophosphonic acids (PFPAs) and fluorotelomer phosphate diesters (di-PAPs) occurred in 100 and 92% of biosolid samples, respectively. Analysis of mass flows showed that, for both quantified (on a fluorine mass basis) and semi-quantified PFAS, the majority of PFAS exited WWTPs through the aqueous effluent compared to the biosolids stream. Overall, these results highlight the importance of semi-quantified PFAS precursors in WWTPs, and the need to further understand the impacts of their ultimate fate in the environment.

Metagenomic surveillance of antibiotic resistome in influent and effluent of wastewater treatment plants located on the Qinghai-Tibetan Plateau

As hotspots for the dissemination of antibiotic resistance genes (ARGs), wastewater treatment plants (WWTPs) have attracted global attention. However, there lacks a sufficient metagenomic surveillance of antibiotic resistome in the WWTPs located on the Qinghai-Tibet Plateau. Here, metagenomic approaches were used to comprehensively investigate the occurrence, mobility potential, and bacterial hosts of ARGs in influent and effluent of 18 WWTPs located on the Qinghai-Tibet Plateau. The total ARG relative abundances and diversity were significantly decreased from influent to effluent across the WWTPs. Multidrug, bacitracin, sulfonamide, aminoglycoside, and beta-lactam ARGs generally consisted of the main ARG types in effluent samples, which were distinct from influent samples. A group of 72 core ARGs accounting for 61.8–95.8 % of the total ARG abundances were shared by all samples. Clinically relevant ARGs mainly conferring resistance to beta-lactams were detected in influent (277 ARGs) and effluent (178 ARGs). Metagenomic assembly revealed that the genetic location of an ARG on a plasmid or a chromosome was related to its corresponding ARG type, demonstrating the distinction in the mobility potential of different ARG types. The abundance of plasmid-mediated ARGs accounted for a much higher proportion than that of chromosome-mediated ARGs in both influent and effluent. Moreover, the ARGs co-occurring with diverse mobile genetic elements in the effluent exhibited a comparable mobility potential with the influent. Furthermore, 137 metagenome-assembled genomes (MAGs) assigned to 13 bacterial phyla were identified as the ARG hosts, which could be effectively treated in most WWTPs. Notably, 46 MAGs were found to carry multiple ARG types and the potential pathogens frequently exhibited multi-antibiotic resistance. Some ARG types tended to be carried by certain bacteria, showing a specific host-resistance association pattern. This study highlights the necessity for metagenomic surveillance and will facilitate risk assessment and control of antibiotic resistome in WWTPs located on the vulnerable area.

Wastewater treatment plants discharges disseminated more Giardia than Cryptosporidium.

Parasitic protozoa Giardia intestinalis and Cryptosporidium parvum are causative agents for giardiasis and cryptosporidiosis, respectively. These infections are mostly associated with waterborne diseases. The discharges from wastewater treatment plants (WWTPs) that reach surface waters cause waterborne transmission because there are no regulations for monitoring these protozoa. This emphasizes how crucial the removal capacities of WWTPs to prevent the spread of infectious parasitic pathogens. For this reason, in this study, five different types of WWTPs including conventional activated sludge (CAS), biological nutrient removal (BNR), sequencing batch reactor (SBR), membrane bioreactor (MBR), and WWTP with coagulation-flocculation and UV disinfection (CoFlUV) units were investigated over a year, seasonally in terms of their G. intestinalis and C. parvum removal capacities. The seasonal abundances of these protozoa-specific genes in both the influents and effluents of each WWTP were determined by qPCR. The reduction of protozoan rDNA copies in the effluent wastewater samples compared with the influent wastewater samples was assessed as log10 reduction values (LRVs). LRVs >3 were reachable for C. parvum in all types of WWTPs tested. However, only LRVs 1-2 were reachable for G. intestinalis in CAS, SBR, CoFlUV, and MBR. Significant seasonal variations were just observed in SBR and CAS for G. intestinalis and C. parvum (p < 0.05), respectively. The findings depicted that WWTPs tested disseminated more giardiasis causative agents than cryptosporidiosis. Therefore, G. intestinalis needs to be monitored in WWTPs' discharges to reduce any potential damage of this parasite to public health. PRACTITIONER POINTS: Removal of G. intestinalis and C. parvum in WWTPs was affected by the process. LRV 2.92 was the highest LRV achieved for G. intestinalis. LRV >3 was reachable for C. parvum. WWTPs discharges disseminated more G. intestinalis than C. parvum. WWTPs effluents should be monitored in terms of G. intestinalis.

Extraction of selected benzothiazoles, benzotriazoles and benzenesulfonamides from environmental water samples using a home-made sol-gel silica-based mixed-mode zwitterionic sorbent modified with graphene

A novel sol-gel silica-based mixed-mode zwitterionic sorbent modified with graphene microparticles was synthesized. Thanks to the inclusion of multiple functional groups and graphene microparticles to exert a wide range of intermolecular/interionic interactions including dipole-dipole interactions, ion-exchange interactions and π-π interactions, the sorbent showed high retention in the solid-phase extraction (SPE) of benzothiazoles, benzotriazoles and benzenesulfonamides.The SPE protocol was optimized in terms of pH, sample loading volume and elution conditions using liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS). The method based on SPE followed by LC-HRMS was validated. Apparent recoveries at two levels of concentration were in the range from 48 to 85% (in most cases) in matrices such as influent wastewater, matrix effect was lower than ±30% in most cases, method detection and quantification limits being lower than 20 ng/L and repeatability and reproducibility between days were lower than 18% (n = 4).River, effluent and influent wastewaters samples were analyzed, obtaining concentrations ranging from 3 to 175 ng/L in river samples, from 12 to 499 ng/L in effluent samples and from 15 to 632 ng/L in influent samples, when the compounds were above the method quantification limits.

Unveiling the Occurrence and Potential Ecological Risks of Organophosphate Esters in Municipal Wastewater Treatment Plants across China.

Organophosphate esters (OPEs) discharged from wastewater treatment plants (WWTPs) have attracted increasing concerns because of their potential risks to aquatic ecosystems. The identification of the structures of OPEs is a prerequisite for subsequent assessment of their environmental impacts, which could hardly be accomplished using traditional target analytical methods. In this study, we describe the use of suspect and nontarget screening techniques for identification of organophosphate triesters and diesters (tri-OPEs and di-OPEs) in the influent and effluent samples acquired from 25 municipal WWTPs across China. There are totally 33 different OPE molecules identified, 11 of which are detected in wastewater for the first time and 4 are new to the public. In all tested samples, di-OPEs account for a significant portion (53% on average) of the total OPEs (ng/L-μg/L). More importantly, most of the OPEs could not be eliminated after treatment in these WWTPs, while some of the di-OPEs even accumulate. The research priority of OPEs in the effluent based on ecological risk was also analyzed, and the results reflected a previously unrecognized exposure risk of emerging OPEs for aquatic living organisms. These findings present a holistic understanding of the environmental relevance of OPEs in WWTPs on a country scale, which will hopefully provide guidance for the upgrade of treatment protocols in WWTPs and even for the modification of governmental regulations in the future.

Effective removal of trace 1,4-dioxane by biological treatments augmented with propanotrophic single culture versus synthetic consortium

1,4-Dioxane (dioxane) is historically used as a stabilizer for chlorinated solvents and has emerged with increasing attention due to its frequent detection at numerous sites in the US. However, trace concentrations of dioxane and intricate environmental conditions hinder its effective treatment in the field. A number of propanotrophic microorganisms are reputed for their ability to degrade dioxane via metabolism (e.g., Pseudonocardia dioxanivorans CB1190 and Mycobacterium dioxanotrophicus PH-06) and cometabolism (e.g., Azoarcus sp. DD4). In this study, we assessed the feasibility of the novel dioxane cometabolizer, Azoarcus sp. DD4, and the mixture of CB1190, PH-06, and DD4 at the ratio of 1:1:2 to remove dioxane in various water samples from a landfill site in northern New Jersey. After being fed with propane for 6∼9 days, DD4 or the synthetic consortium can effectively degrade dioxane to below our method detection limit (i.e., 0.38 µg/L) in both influent and effluent samples from the existing pump-and-treat facility, as well as the contaminated groundwater from the monitoring well near the source zone of the plume. Relative abundances of Azoarcus and DD4’s toluene monooxygenase gene tmoA were both positively correlated with dioxane degradation rates observed in microcosms, suggesting DD4’s pivotal contribution. In light of both qPCR and 16S rRNA amplicon sequencing analyses, DD4 remained dominant (29∼52%) in these three types of groundwater, indicating DD4’s compatibility for both in situ and ex situ treatments. Interestingly, inoculation with the synthetic consortium did not elicit significant improvement in dioxane removal as compared with the treatments seeded with DD4 alone. Unlike DD4, CB1190 and PH-06 were absent after the complete course of the microcosm study, indicating the decay of both two exogenous species. This was probably due to outcompetion by DD4 and/or indigenous microbiomes and hindrance by field factors (e.g., inhibitors and nutrients). Native Rhodococcus species emerged and exhibited significant correlation with the prmA gene encoding the alpha subunit of propane monooxygenase of PH-06, suggesting them as possible recipients of this dioxane degradation gene via horizontal transfer. Collectively, this study demonstrated the DD4’s high viability and excellent compatibility with diverse field samples suited as an effective inoculum to mitigate trace levels of dioxane in the subsurface or in bioreactors. Amendment of Gram-positive propanotrophic metabolizers has the potential to spread dioxane degradation genes though their viability can be low at certain sites.

Multi-class determination of pharmaceuticals as emerging contaminants in wastewater from Eastern Province, Saudi Arabia using eco-friendly SPE-UHPLC-MS/MS: Occurrence, removal and environmental risk assessment

Occurrence of pharmaceuticals in wastewater, which might be released to different water environments causing contamination might lead to health hazards. Therefore, sensitive, selective and reliable analytical methods are required to monitor pharmaceuticals in such complex environmental matrix. In this work, an eco-friendly, sensitive, robust and selective ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) method was developed to simultaneously identify and quantify multiple classes of pharmaceuticals in influent and effluent wastewater samples in Eastern Province, Saudi Arabia using solid phase extraction (SPE) for sample preparation. The extraction conditions were optimized to achieve the best extraction efficiencies and achieved recoveries of 70.4 – 112.2% and 74.0 – 107.8% in influent (IWW) and effluent wastewater (EWW) samples, respectively. The optimized method had satisfactory precision represented as %RSD lower than 13.04%. Limits of detection were 0.06 – 16.0 and 0.07 – 13.23 ng L-1 for IWW and EWW, respectively. The developed method was successfully employed for the analysis of 32 pharmaceuticals in wastewater. The greenness profile was assessed using two assessment tools; Analytical Eco-scale and AGREE metric. Caffeine and Ciprofloxacin were the most frequent and highest concentration in influent wastewater with concentration range 23,600 – 37,602 ng L-1 and 1,684 –3,585 ng L-1, respectively. Average effluent concentrations range was from below method quantification limit to 1,241 ng L-1 (ciprofloxacin). Removal efficiency ranged from - 3.1% (trimethoprim) to 99% (caffeine). Environmental risk assessment was calculated using risk quotient approach for each individual analyte and for the mixture. The results revealed 4 and 6 compounds with high and medium risk, respectively. This work provides new insights into the occurrence, removal and ecotoxicological risk of pharmaceuticals from wastewater in Eastern Province, Saudi Arabia. It emphasizes the importance of monitoring these compounds as an input for future mitigation actions.

Community-Scale Wastewater Surveillance of Candida auris during an Ongoing Outbreak in Southern Nevada

Candida auris is an opportunistic fungal pathogen and an emerging global public health threat, given its high mortality among infected individuals, antifungal resistance, and persistence in healthcare environments. This study explored the applicability of wastewater surveillance for C. auris in a metropolitan area with reported outbreaks across multiple healthcare facilities. Influent or primary effluent samples were collected over 10 weeks from seven sewersheds in Southern Nevada. Pelleted solids were analyzed using an adapted quantitative polymerase chain reaction (qPCR) assay targeting the ITS2 region of the C. auris genome. Positive detection was observed in 72 of 91 samples (79%), with higher detection frequencies in sewersheds serving healthcare facilities involved in the outbreak (94 vs 20% sample positivity). Influent wastewater concentrations ranged from 2.8 to 5.7 log10 gene copies per liter (gc/L), and primary clarification achieved an average log reduction value (LRV) of 1.24 ± 0.34. Presumptive negative surface water and wastewater controls were non-detect. These results demonstrate that wastewater surveillance may assist in tracking the spread of C. auris and serve as an early warning tool for public health action. These findings provide the foundation for future application of wastewater-based epidemiology (WBE) to community- or facility-level surveillance of C. auris and other high consequence, healthcare-associated infectious agents.

National Wastewater Reconnaissance of Analgesic Consumption in Australia.

A wastewater-based epidemiology (WBE) method is presented to estimate analgesic consumption and assess the burden of treated pain in Australian communities. Wastewater influent samples from 60 communities, representing ∼52% of Australia's population, were analyzed to quantify the concentration of analgesics used to treat pain and converted to estimates of the amount of drug consumed per day per 1000 inhabitants using pharmacokinetics and WBE data. Consumption was standardized to the defined daily dose per day per 1000 people. The population burden of pain treatment was classified as mild to moderate pain (for non-opioid analgesics) and strong to severe pain (for opioid analgesics). The mean per capita weighted total DDD of non-opioid analgesics was 0.029 DDD/day/person, and that of opioid-based analgesics was 0.037 DDD/day/person across Australia. A greater burden of pain (mild to moderate or strong to severe pain index) was observed at regional and remote sites. The correlation analysis of pain indices with different socioeconomic descriptors revealed that pain affects populations from high to low socioeconomic groups. Australians spent an estimated US $3.5 (AU $5) per day on analgesics. Our findings suggest that WBE could be an effective surveillance tool for estimating the consumption of analgesics at a population scale and assessing the total treated pain burden in communities.

Direct wastewater extraction as a simple and effective method for SARS-CoV-2 surveillance and COVID-19 community-level monitoring.

Wastewater surveillance has proven to be an effective tool to monitor the transmission and emergence of infectious agents at a community scale. Workflows for wastewater surveillance generally rely on concentration steps to increase the probability of detection of low-abundance targets, but preconcentration can substantially increase the time and cost of analyses while also introducing additional loss of target during processing. To address some of these issues, we conducted a longitudinal study implementing a simplified workflow for SARS-CoV-2 detection from wastewater, using a direct column-based extraction approach. Composite influent wastewater samples were collected weekly for 1 year between June 2020 and June 2021 in Athens-Clarke County, Georgia, USA. Bypassing any concentration step, low volumes (280µl) of influent wastewater were extracted using a commercial kit, and immediately analyzed by RT-qPCR for the SARS-CoV-2 N1 and N2 gene targets. SARS-CoV-2 viral RNA was detected in 76% (193/254) of influent samples, and the recovery of the surrogate bovine coronavirus was 42% (IQR: 28%, 59%). N1 and N2 assay positivity, viral concentration, and flow-adjusted daily viral load correlated significantly with per-capita case reports of COVID-19 at the county-level (ρ = 0.69-0.82). To compensate for the method's high limit of detection (approximately 106-107 copies l-1 in wastewater), we extracted multiple small-volume replicates of each wastewater sample. With this approach, we detected as few as five cases of COVID-19 per 100000 individuals. These results indicate that a direct-extraction-based workflow for SARS-CoV-2 wastewater surveillance can provide informative and actionable results.

The removal of ciprofloxacin from synthetic wastewater in constructed wetland

Considering the health effects of antibiotics in the environment, effective monitoring and treatment technologies are needed to mitigate social and environmental impacts. The present study was carried out to investigate the efficiency of the constructed wetland (CW) on the removal of Ciprofloxacin (CIP) from aqueous samples. Experiments were conducted in pilot scale CWs planted with single plants of Cyperus alternifolius, Canna indica and one planted with both plant species. Analysis of CIP concentrations in the influent and effluent samples was done using Cary 60 UV–Vis spectrophotometer, while physical-chemical parameters were monitored for the influent and effluent samples. The removal efficiency of physico-chemical parameters was ˃70% for Nitrate, ˃60% for Phosphate, ˃70% for BOD and ˃77% for COD. The maximum removal of CIP (77.1%) was observed in CW planted with Cyperus alternifolius during a 7 days hydraulic retention time (HRT). The results of this study show superior performance of Cyperus alternifolius than Canna indica. There was no significance difference (p > 0.05) produced by mixing the two plants in a CW. However, mixing of plants especially ornamental plants in CWs brings good visual impression of the systems while treating the wastewater. This study demonstrate that CW can remove antibiotics from wastewater. The best performance depends on best selection and best combination of the plants.

Removal of arsenic in a sand filter coupled with zero valent iron

Arsenic (As) in wastewater has negative effects on the environment and human health, hence As containing wastes must be handled properly. Given the accessibility of metallic iron, studies investigating into the potential application of zerovalent iron in the removal of arsenic are promising. In this study, the performance of sand filter blended with several kinds of zero valent iron (Fe0), such as iron wool, iron fillings, and iron nails, were compared. These materials were combined in a sand filter column, and the efficiency was calculated using the As concentrations in the influent and effluent samples. Experiments were carried out in order to compare performance as a function of Fe0 dose and contact time. The outcome of this investigation showed that sand filter containing iron wool had a better removal efficiency of arsenic removal than iron filings and iron nails. The results in all columns showed that as dosage was increased, removal efficiency of arsenic increased significantly. In case of contact time the results revealed that arsenic can effectively be removed from water in the first 48 h. The early adsorption response is quick in all columns, but get slower as time goes on. The highest removal efficiency was 99.6% and the lowest removal efficiency was 82.7%.