Raw Wastewater Samples Research Articles

Antibiotic resistance genes and crAssphage in hospital wastewater and a canal receiving the treatment effluent

Hospital wastewater is a major hotspot for the spread of antimicrobial resistance (AMR) in aquatic ecosystems. This study aimed to investigate the prevalence of antibiotic resistance genes (ARGs) and their correlation with crAssphage in a hospital wastewater treatment plant (HWWTP) and a receiving canal. Water samples were analyzed for 94 ARGs and crAssphage relative to the 16S rRNA using high-throughput quantitative polymerase chain reaction (HT-qPCR). Subsequently, 7 ARGs and crAssphage were selected and quantified using qPCR. The results showed that the detected genes ranged from 79 to 93 out of 95 genes. The raw wastewater (WW) samples had the highest gene diversity compared to the upstream canal, which had less diversity than downstream samples, as determined by HT-qPCR. The blaGES was the most abundant in WW samples, while qacEΔ1, merA, IS6100, tnpA, and IS26 showed high prevalence throughout the treatment processes. The concentrations of intI1, sul1, blaTEM,blaNDM,blaVIM,tetQ, mcr-1, crAssphage, and 16S rRNA, measured using qPCR, were the highest in WW and significantly reduced in treated water samples. Although some water quality parameters, such as total suspended solids and dissolved oxygen, did not significantly differ before and after treatment, removal efficiency ranged from 0.60 to 3.23 log reduction values (LRV). The highest LRV was observed for the tetQ, whereas the mcr-1 had the lowest LRV. Strong positive correlations among the absolute concentrations of ARGs and crAssphage were observed (Spearman's rho = 0.6–1.0), and biochemical oxygen demand correlated with blaTEM and blaVIM (Spearman's rho = 0.6). These results indicate that crAssphage and water quality could reflect the distribution of other ARGs throughout the HWWTP. Further studies are needed to underscore the importance of monitoring ARGs and genetic markers such as crAssphage in HWWTPs and their receiving waters to enhance our understanding of ARG distribution.

Implications of long-term sample storage on the recovery of viruses from wastewater and biobanking

Wastewater-based monitoring has been widely implemented worldwide for the tracking of SARS-CoV-2 outbreaks and other viral diseases. In many surveillance programmes, unprocessed and processed wastewater samples are often frozen and stored for long periods of time in case the identification and tracing of an emerging health threat becomes necessary. However, extensive sample bioarchives may be difficult to maintain due to limitations in ultra-freezer capacity and associated cost. Furthermore, the stability of viruses in such samples has not been systematically investigated and hence the usefulness of bioarchives is unknown. In this study, we assessed the stability of SARS-CoV-2, influenza viruses, noroviruses and the faecal indicator virus, crAssphage, in raw wastewater and purified nucleic aacid extracts stored at -80 °C for 6–24 months. We found that the isolated viral RNA and DNA showed little signs of degradation in storage over 8–24 months, whereas extensive decay viral and loss of qPCR signal was observed during the storage of raw unprocessed wastewater. The most stable viruses were noroviruses and crAssphage, followed by SARS-CoV-2 and influenza A virus. Based on our findings, we conclude that bioarchives comprised of nucleic acid extracts derived from concentrated wastewater samples may be archived long-term, for at least two years, whereas raw wastewater samples may be discarded after one year.

Combined electrocoagulation/flotation technique and membrane desalination for textile wastewater reuse

A novel integrated system that combined electrocoagulation/flotation (ECF) technique with membrane desalination was used for textile wastewater treatment to reduce the environmental pollution of textile wastewater. Two scenarios were examined: the first is ECF followed by membrane desalination; the second is the application of membrane desalination for treatment of raw wastewater. Iron electrodes were used in batch-wise tests to examine the effects of electrolysis time and current intensity on percentage removals. The SiO2/PA(TFC) membrane was used to treat raw wastewater and ECF effluents. As the current intensity increased from 50 mA to 600 mA, the color elimination was improved from 94 % to 99 % and the COD elimination improved from 59.1 % to 81.5 %. The findings demonstrate the efficacy of ECF in removing color and COD, although the treated wastewater sample's TDS increased from 4200 mg/L to 19,400 mg/L (raw textile wastewater sample). The SiO2/PA(TFC) membrane displays the salt rejection and water flux reduction of the raw wastewater samples 95 %, and 22 (L/m2.h). Corresponding results for ECF treated wastewater were 94, 93, 91 % and 13, 11.5, and 10 (L/m2.h), respectively. The SiO2/PA(TFC) membrane displays the reduction of COD from 760 mg O2/L (raw) and 310 mg O2/L (ECF effluent) to zero%. Also, it proves the capabilities of SiO2/PA(TFC) membrane for the removal of TDS, COD and color. This integrated system can provide sustainable source for fresh water supply used for landscape, irrigation, industry and various purposes.

Wastewater-based epidemiology methodology to investigate human exposure to bisphenol A, bisphenol F and bisphenol S.

Wastewater-based epidemiology (WBE) has become an invaluable tool for tracking the evolution of use or exposure of/to numerous substances. Bisphenols, commonly utilized in manufacturing plastic goods, have been categorized as endocrine disrupting chemicals, underscoring the critical need for real-time data on their local-level exposure to safeguard public health. In this study, we have developed a novel analytical method and WBE framework for the assessment of population-level exposure to bisphenol A (BPA) and its most prominent substitutes, bisphenols F and S (BPF and BPS), through the determination their Phase II metabolites in wastewater by WBE. Stability and exclusivity tests denoted that glucuronides are not stable in sewage, whereas sulfate metabolites are good biomarkers. Therefore, a solid-phase extraction followed by liquid chromatography-tandem mass spectrometry method was developed for the bisphenols’ monosulfates and BPA bissulfate. The analytical method was validated with three different wastewater matrices, providing trueness (as recovery) in the 79–112 % range with relative standard deviations < 12 %, and method quantification limits below 2 ng L−1 for monosulfates, but higher (35 ng L−1) for BPA bissulfate. Subsequently, the method was applied to 24h-composite raw wastewater samples collected over a week in 4 different locations in Spain and Portugal. BPA bissulfate was not detected, but the three monosulfate metabolites of each bisphenol were positively detected in the samples, being the metabolite of BPA the most prevalent, followed by those of BPF and BPS. Community-wide BPA intake was then estimated to be higher than the European Food Safety Agency (EFSA) tolerable daily intake (TDI) of 2 × 10−4 µg kg−1day−1 in all locations. In the case of BPF and BPS, there is not enough metabolism data or even established limit, but they would also surpass safe levels in several locations if a similar metabolism and TDI would be assumed. This innovative method could be used to a larger set of wastewater-treatment plants as an early-warning approach on human exposure to bisphenols.

Integrated hepatitis e virus monitoring in central Argentina: a six-year analysis of clinical surveillance and wastewater-based epidemiology

Wastewater-based epidemiology (WBE) has gained prominence worldwide as a powerful tool in public health. This study aimed to monitor the circulation of Hepatitis E Virus (HEV) from wastewater samples collected during a six-year period and compare these results with clinical surveillance in the central region of Argentina. From 2017 to 2022, 1008 raw wastewater samples were analyzed, including four wastewater treatment plants from four cities (n=319), and 7 local neighborhood collector sewers in Córdoba city (n=689). Serum and/or stool samples from patients suspected of HEV infection were also analyzed (n=48). HEV molecular detection and viral load quantification were performed by real time RT-qPCR, and genetic characterization by two RT-Nested PCRs (targeting partial ORF-1 and ORF-2 genomic regions), sequencing and phylogenetic analysis.Fifty-three (5.3%) wastewater samples were RNA-HEV positive by real time RT-qPCR, with variations according to the location and year (0.0% - 21.6%). Out of these, ORF-2 genomic region was amplified in 20 samples (37.7%) and ORF-1 partial region in 12 (22.6%), and eighteen sequences were obtained. Throughout the study period, two (4.2%) HEV confirmed infections were reported, and one sequence was obtained. Phylogenetic analyses for both genomic regions showed that all the isolates were genotype HEV-3 clade abchijklm.Our study detected HEV in wastewater over a six-year period, despite a low number of clinical cases, emphasizing WBE as a valuable tool that complements clinical surveillance, by detecting pathogens' presence; identifying their transmission, circulation dynamics and excretion hotspots; and revealing changes in their genomic diversity.

Assessing the prevalence of FRNA bacteriophages and their correlation with SARS-CoV-2 RNA in the wastewater of Mumbai city

ABSTRACT Bacteriophages have been explored as indicators of the presence of human viruses in water. F+ coliphages are of particular interest due to their abundance in wastewater and some groups surviving the treatment process. This study assessed the prevalence of F+ coliphages and FRNA bacteriophage (FRNAPH) groups in the wastewaters of Mumbai city and explored its correlation with the presence of SARS-CoV-2. Wastewater samples (raw = 63, treated = 99) collected from three wastewater treatment plants (WWTPs) were assessed for F+ coliphages, FRNAPH groups and SARS-CoV-2. Of the 63 raw wastewater samples, 92% were positive for F+ coliphages and FRNAPH, while SARS-CoV-2 was detected in 76% of samples; FRNAPH GI RNA was the most prevalent (96.82%) followed by FRNAPH GII (77.77%), FRNAPH GIII (36.50%), and FRNAPH GIV (4.76%). A significant correlation was observed between the occurrence of SARS-CoV-2 and F+ coliphages, FRNAPH GII and FRNAPH GIII RNA copies and also between F+ coliphages and FRNAPH GII and GIII. These observations suggest that F+ coliphages, FRNAPH GII and GIII can be explored further as indicators of SARS-CoV-2 in wastewaters. Also, F+ coliphage detection may be a cost-effective and practical approach for monitoring virus elimination during wastewater treatment.

Dynamics of antimicrobial resistance and susceptibility profile in full-scale hospital wastewater treatment plants.

Drug resistance has become a matter of great concern, with many bacteria now resist multiple antibiotics. This study depicts the occurrence of antibiotic-resistant bacteria (ARB) and resistance patterns in five full-scale hospital wastewater treatment plants (WWTPs). Samples of raw influent wastewater, as well as pre- and post-disinfected effluents, were monitored for targeted ARB and resistance genes in September 2022 and February 2023. Shifts in resistance profiles of Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii antimicrobial-resistant indicators in the treated effluent compared to that in the raw wastewater were also worked out. Ceftazidime (6.78 × 105 CFU/mL) and cefotaxime (6.14 × 105 CFU/mL) resistant species showed the highest concentrations followed by ciprofloxacin (6.29 × 104 CFU/mL), and gentamicin (4.88 × 104 CFU/mL), in raw influent respectively. WWTP-D employing a combination of biological treatment and coagulation/clarification for wastewater decontamination showed promising results for reducing ARB emissions from wastewater. Relationships between treated effluent quality parameters and ARB loadings showed that high BOD5 and nitrate levels were possibly contributing to the persistence and/or selection of ARBs in WWTPs. Furthermore, antimicrobial susceptibility tests of targeted species revealed dynamic shifts in resistance profiles through treatment processes, highlighting the potential for ARB and ARGs in hospital wastewater to persist or amplify during treatment.

Consumption of illicit drugs and benzodiazepines in six Spanish cities during different periods of the COVID-19 pandemic

Wastewater-based epidemiology (WBE) can provide objective and real time information about the use of addictive substances. A national study was conducted by measuring the most consumed illicit drugs, other drugs whose consumption is not so widespread but has increased significantly in recent years, and benzodiazepines in untreated wastewater from seven wastewater treatment plants (WWTPs) in six Spanish cities. Raw composite wastewater samples were collected from December 2020 to December 2021, a period in which the Spanish and regional governments adopted different restriction measures to contain the spread of the COVID-19 pandemic. Samples were analyzed using a validated analytical methodology for the simultaneous determination of 18 substances, based on solid-phase extraction and liquid-chromatography tandem mass spectrometry. Except for heroin, fentanyl, 6-acetylmorphine and alprazolam, all the compounds were found in at least one city and 9 out of 18 compounds were found in all the samples. In general, the consumption of illicit drugs was particularly high in one of the cities monitored in December 2020, when the restrictions were more severe, especially for cannabis and cocaine with values up to 46 and 6.9 g/day/1000 inhabitants (g/day/1000 inh), respectively. The consumption of MDMA, methamphetamine and mephedrone was notably higher in June 2021, after the end of the state of alarm, in the biggest population investigated in this study. Regarding the use of benzodiazepines, the highest mass loads corresponded to lorazepam. This study demonstrates that WBE is suitable for complementing epidemiological studies about the prevalence of illicit drugs and benzodiazepines during the COVID-19 pandemic restrictions.

Analysis of chlorhexidine, antibiotics and bacterial community composition in water environments from Brazil, Cameroon and Madagascar during the COVID-19 pandemic

The widespread of chlorhexidine and antibiotics in the water bodies, which grew during the global COVID-19 pandemic, can increase the dispersion of antibiotic resistance. We assessed the occurrence of these pharmaceutical compounds as well as SARS-CoV-2 and analysed the bacterial community structure of hospital and urban wastewaters from Brazil, Cameroon, and Madagascar. Water and wastewater samples (n = 59) were collected between January–June 2022. Chlorhexidine, azithromycin, levofloxacin, ceftriaxone, gentamicin and meropenem were screened by Ultra-High-Performance Liquid Chromatography coupled with mass spectrometer. SARS-CoV-2 was detected based on the nucleocapsid gene (in Cameroon and Madagascar), and envelope and spike protein-encoding genes (in Brazil). The total community-DNA was extracted and used for bacterial community analysis based on the 16S rRNA gene. To unravel likely interaction between pharmaceutical compounds and/or SARS-CoV-2 with the water bacterial community, multivariate statistics were performed. Chlorhexidine was found in hospital wastewater effluent from Brazil with a maximum concentration value of 89.28 μg/L. Additionally, antibiotic residues such as azithromycin and levofloxacin were also present at concentrations between 0.32–7.37 μg/L and 0.11–118.91 μg/L, respectively. In Cameroon, azithromycin was the most found antibiotic present at concentrations from 1.14 to 1.21 μg/L. In Madagascar instead, ceftriaxone (0.68–11.53 μg/L) and levofloxacin (0.15–0.30 μg/L) were commonly found. The bacterial phyla statistically significant different (P < 0,05) among participating countries were Proteobacteria, Patescibacteria and Dependentiae which were mainly abundant in waters sampled in Africa and, other phyla such as Firmicutes, Campylobacterota and Fusobacteriota were more abundant in Brazil. The phylum Caldisericota was only found in raw hospital wastewater samples from Madagascar. The canonical correspondence analysis results suggest significant correlation of azithromycin, meropenem and levofloxacin with bacteria families such as Enterococcaceae, Flavobacteriaceae, Deinococcaceae, Thermacetogeniaceae and Desulfomonilaceae, Spirochaetaceae, Methanosaetaceae, Synergistaceae, respectively. Water samples were also positive for SARS-CoV-2 with the lowest number of hospitalized COVID-19 patients in Madagascar (n = 7) and Brazil (n = 30). Our work provides new data about the bacterial community profile and the presence of pharmaceutical compounds in the hospital effluents from Brazil, Cameroon, and Madagascar, whose limited information is available. These compounds can exacerbate the spreading of antibiotic resistance and therefore pose a risk to public health.

Exposure to hull cleaning wastewater induces mortality through oxidative stress and cholinergic disturbance in the marine polychaete Perinereis aibuhitensis

While wastewater and paint particles discharged from the in-water cleaning process of ship hulls are consistently released into benthic ecosystems, their hazardous effects on non-target animals remain largely unclear. In this study, we provide evidence on acute harmful effects of hull cleaning wastewater in marine polychaete Perinereis aibuhitensis by analyzing physiological and biochemical parameters such as survival, burrowing activity, and oxidative status. Raw wastewater samples were collected during ship hull cleaning processes in the field. Two wastewater samples for the exposure experiment were prepared in the laboratory: 1) mechanically filtered in the in-water cleaning system (MF) and 2) additionally filtered with a 0.45 μm filter in the laboratory (LF). These wastewater samples contained high concentrations of metals (zinc and copper) and metal–based booster biocides (copper pyrithione and zinc pyrithione) compared to those analyzed in seawater. Polycheates were exposed to different concentrations of the two wastewater samples for 96 h. Higher mortality was observed in response to MF compared to LF–exposed polychaetes. Both wastewater samples dose-dependently decreased burrowing activity and AChE activity. Drastic oxidative stress was observed in response to the two wastewater samples. MDA levels were significantly increased by MF and LF samples. Significant GSH depletion was observed with MF exposure, while increased and decreased GSH contents were observed in LF-exposed polychaetes. Enzymatic activities of antioxidant components, catalase, superoxide dismutase, and glutathione S-transferase were significantly modulated by both wastewater samples. These results indicate that even filtered hull cleaning wastewater can have deleterious effects on the health status of polychaetes.

Multidrug-Resistant Staphylococcus sp. and Enterococcus sp. in Municipal and Hospital Wastewater: A Longitudinal Study.

The objective of the study was to detect multidrug-resistant Staphylococcus sp. and Enterococcus sp. isolates in municipal and hospital wastewater and to determine their elimination or persistence after wastewater treatment. Between August 2021 and September 2022, raw and treated wastewater samples were collected at two hospital and two community wastewater treatment plants (WWTPs). In each season of the year, two treated and two raw wastewater samples were collected in duplicate at each of the WWTPs studied. Screening and presumptive identification of staphylococci and enterococci was performed using chromoagars, and identification was performed with the Matrix Assisted Laser Desorption Ionization Time of Flight mass spectrometry (MALDI-TOF MS®). Antimicrobial susceptibility was performed using VITEK 2® automated system. There were 56 wastewater samples obtained during the study period. A total of 182 Staphylococcus sp. and 248 Enterococcus sp. were identified. The highest frequency of Staphylococcus sp. isolation was in spring and summer (n = 129, 70.8%), and for Enterococcus sp. it was in autumn and winter (n = 143, 57.7%). Sixteen isolates of Staphylococcus sp. and sixty-three of Enterococcus sp. persisted during WWTP treatments. Thirteen species of staphylococci and seven species of enterococci were identified. Thirty-one isolates of Staphylococcus sp. and ninety-four of Enterococcus sp. were multidrug-resistant. Resistance to vancomycin (1.1%), linezolid (2.7%), and daptomycin (8.2%/10.9%%), and a lower susceptibility to tigecycline (2.7%), was observed. This study evidences the presence of Staphylococcus sp. and Enterococcus sp. resistant to antibiotics of last choice of clinical treatment, in community and hospital wastewater and their ability to survive WWTP treatment systems.

Are lagoons efficient for summer wastewater treatment in northern decentralized communities to ensure environmental and human health?

Among wastewater treatment methods, wastewater treatment lagoons are popular in northern decentralized regions. There is however a shortage of studies on northern wastewater treatment efficiency. This research addresses this gap by sequentially evaluating lagoon efficiency in a northern Canadian community, aiming to determine the feasibility or the necessity for improvement or alternative methods. The study encompasses physicochemical and human enteric viruses' detection in wastewater, crucial for mitigating environmental and public health risks. Wastewater lagoons exhibited removal efficiencies of 94.10 % for total suspended solids (TSS), 70.5 % for total solids (TS), 99.89 % for ammonium (NH4), 61.53 % for total nitrogen (TN), 65.85 % for phosphate (PO43−), and 91.3 % for chemical oxygen demand (COD). Human noroviruses GI and GII were detected in raw wastewater and lagoon samples, indicating their presence in the community's sewage system. The inefficiency of the lagoon system in reducing TSS, COD, and PO43− to below standard levels, along with the presence of viruses, raises concerns for environmental and human health in the region, especially considering the community's heavy dependence on land and water resources. Urgent measures are imperative to address these inefficiencies for the well-being of both the environment and the community.

Remediation of Brewery Wastewater Using Green Synthesized Nano-Particles

The brewing industry consumes a large amount of water needed for brewing, rinsing, and cooling purposes, and therefore produces huge amount of effluents. Therefore, the objective of this paper was to evaluate the use of Moringa oleiferra (MO) powder and synthesized 1.0 and 2.0 g TiO2NPs as green synthesized nano-particles for the remediation of brewery wastewater using standard methods. The raw wastewater sample characterization for pH, BOD, COD, Lead and coliform count were: 7.26, 935, 1045, 0.083 mg/L and 136 cfu/100 mL respectively. Results of the UV – Visible spectrophotometer showed the maximum wavelength of 275, 275, 278 and 282.50 nm for 5:20, 10:20, 15:20, 20:20 of MO and TiO2 ratio respectively, while the FTIR results show the presence of oxygen surface complex groups such as hydroxyl and carbonyl. The SEM reveals a porous surface area accompanied by several wide opening pores of different sizes and shapes, while EDX shows the concentration of titanium, Sulphur and silicon in percent weight; 85.79, 2.96 and 1.46 % respectively. Results of the wastewater treated with 50 g defatted M. oleiferra revealed the removal efficiency of 47, 93.2, 56.2, 18, 31.3, 97, 76.1, 81 and 71% for Turbidity, COD, EC, Nitrate, Nitrite, BOD, TS, TDS and TSS respectively. Results of wastewater treated with 1.0 g of TiO2 NPs showed the removal efficiency of 97.8, 94.64, 53.5, 34.2 and 35.1% for COD, BOD, EC, Nitrate and Nitrite respectively. That of 2.0 g of TiO2 NPs showed the removal efficiency of 67, 58, and 87% for Cu, Pb, and Ag respectively. Conclusively, M. oleiferra and varying proportions of green synthesized TiO2 NPs were effective in the remediation of the wastewater from brewery industry as it improves its physicochemical properties, but not so much for the heavy metal concentration.

Real-Time On-Site Monitoring of Viruses in Wastewater Using Nanotrap® Particles and RICCA Technologies.

Wastewater-based epidemiology (WBE) is an effective and efficient tool for the early detection of infectious disease outbreaks in a community. However, currently available methods are laborious, costly, and time-consuming due to the low concentration of viruses and the presence of matrix chemicals in wastewater that may interfere with molecular analyses. In the present study, we designed a highly sensitive "Quick Poop (wastewater with fecal waste) Sensor" (termed, QPsor) using a joint approach of Nanotrap microbiome particles and RICCA (RNA Isothermal Co-Assisted and Coupled Amplification). Using QPsor, the WBE study showed a strong correlation with standard PEG concentrations and the qPCR technique. Using a closed format for a paper-based lateral flow assay, we were able to demonstrate the potential of our assay as a real-time, point-of-care test by detecting the heat-inactivated SARS-CoV-2 virus in wastewater at concentrations of 100 copies/mL and within one hour. As a proof-of-concept demonstration, we analyzed the presence of viral RNA of the SARS-CoV-2 virus and PMMoV in raw wastewater samples from wastewater treatment plants on-site and within 60 min. The results show that the QPsor method can be an effective tool for disease outbreak detection by combining an AI-enabled case detection model with real-time on-site viral RNA extraction and amplification, especially in the absence of intensive clinical laboratory facilities. The lab-free, lab-quality test capabilities of QPsor for viral prevalence and transmission in the community can contribute to the efficient management of pandemic situations.

Enterococci, Van Gene-Carrying Enterococci, and Vancomycin Concentrations in the Influent of a Wastewater Treatment Plant in Southeast Germany.

Vancomycin-resistant (VR) Enterococcus spp. can be detected in high concentrations in wastewaters and pose a risk to public health. During a one-year study (September 2022-August 2023), 24 h composite raw wastewater samples (n = 192) of a municipal wastewater treatment plant were investigated for cultivable enterococci. After growth on Slanetz-Bartley agar (SBA), a mean concentration of 29,736 ± 9919 cfu/mL was calculated. Using MALDI-TOF MS to characterize randomly picked colonies (n = 576), the most common species were found to be Enterococcus faecium (72.6%), E. hirae (13.7%), and E. faecalis (8.0%). Parallel incubation of wastewater samples on SBA and VRESelect agar resulted in a mean rate of VR enterococci of 2.0 ± 1.5%. All the tested strains grown on the VRESelect agar (n = 172) were E. faecium and carried the vanA (54.6%) or vanB gene (45.4%) with limited sequence differences. In susceptibility experiments, these isolates showed a high-level resistance to vancomycin (>256 µg/mL). Concentration of vancomycin was determined in 93.7% of 112 wastewater samples (mean: 123.1 ± 64.0 ng/L) and varied between below 100 ng/L (the detection limit) and 246.6 ng/L. A correlation between the concentration of vancomycin and the rate of VR strains among the total enterococci could not be found. The combination of incubation of samples on SBA and a commercial vancomycin-containing agar applied in clinical microbiology with a multiplex PCR for detection of van genes is an easy-to-use tool to quantify and characterize VR Enterococcus spp. in water samples.

Impact of Corona virus pandemic on wastewater characteristics, treatment, and water reuse in a municipal plant

The COVID-19 pandemic is considered one of the most significant threats to health. The effect of the pandemic on performance of wastewater treatment plants has not received much attention in the literature. This study assessed the impact of Coronavirus pandemic on the wastewater quantity, characteristics, treatment processes, and water reuse in Kuwait. It focused on three municipal wastewater treatment plants (WWTPs) with different design capacities, namely Sulaibiya (600,000 m3/d), Kabd (180,000 m3/d), and Umm Al-Hayman (27,000 m3/d), all using tertiary treatment. Daily data records were obtained on influent and effluent over the years 2018–2020 (before and during the pandemic). In addition, influent and effluent samples were collected from the Sulaibiya and Kabd plants for laboratory analysis to determine the effect of increase in the use of home disinfectants. The results indicate an increase (&amp;lt;50%) in the average daily flow rates received in WWTPs during the 2020 home quarantine lockdowns. Statistical analysis showed stability of the Sulaibiya in BOD5, COD, and TSS removal efficiency while a decrease in the Kabd and UAH plants removal efficiency was observed during the pandemic. During the lockdown period, GC-MS analysis showed a high probability of chloroxylenol (&amp;gt;90), one of the main components of Dettol disinfectant, existing in the raw wastewater samples while the GC-FID analysis indicated the presence of (PAH) as emerging contaminants, at higher concentrations in the influent and effluent samples. The pandemic adversely affected wastewater treatment plant performance and excessive use of Dettol disinfectant by homes resulted in the presence of objectionable organic pollutants in the tertiary-treated effluent which would impair water reuse. Reverse osmosis process used in Sulaibiya plant proved to be effective in removing residual organics and improving effluent quality for reuse during the pandemic.

Candidate biomarkers of antibiotic resistance for the monitoring of wastewater and the downstream environment

Urban wastewater treatment plants (UWTPs) are essential for reducing the pollutants load and protecting water bodies. However, wastewater catchment areas and UWTPs emit continuously antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs), with recognized impacts on the downstream environments. Recently, the European Commission recommended to monitor antibiotic resistance in UWTPs serving more than 100 000 population equivalents. Antibiotic resistance monitoring in environmental samples can be challenging. The expected complexity of these systems can jeopardize the interpretation capacity regarding, for instance, wastewater treatment efficiency, impacts of environmental contamination, or risks due to human exposure. Simplified monitoring frameworks will be essential for the successful implementation of analytical procedures, data analysis, and data sharing. This study aimed to test a set of biomarkers representative of ARG contamination, selected based on their frequent human association and, simultaneously, rare presence in pristine environments. In addition to the 16S rRNA gene, ten potential biomarkers (intI1, sul1, ermB, ermF, aph(3′’)-Ib, qacEΔ1, uidA, mefC, tetX, and crAssphage) were monitored in DNA extracts (n = 116) from raw wastewater, activated sludge, treated wastewater, and surface water (upstream and downstream of UWTPs) samples collected in the Czech Republic, Denmark, Israel, the Netherlands, and Portugal. Each biomarker was sensitive enough to measure decreases (on average by up to 2.5 log-units gene copy/mL) from raw wastewater to surface water, with variations in the same order of magnitude as for the 16S rRNA gene. The use of the 10 biomarkers allowed the typing of water samples whose origin or quality could be predicted in a blind test. The results show that, based on appropriate biomarkers, qPCR can be used for a cost-effective and technically accessible approach to monitoring wastewater and the downstream environment.

Long-term monitoring of COVID-19 prevalence in raw and treated wastewater in Salvador, the largest capital of the Brazilian Northeast

Wastewater-based epidemiology (WBE) becomes an interesting epidemiological approach to monitoring the prevalence of SARS-CoV-2 broadly and non-invasively. Herein, we employ for the first time WBE, associated or not with the PEG 8000 precipitation method, for the detection of SARS-CoV-2 in samples of raw or treated wastewater from 22 municipal wastewater treatment stations (WWTPs) located in Salvador, the fourth most populous city in Brazil. Our results demonstrate the success of the application of WBE for detecting SARS-CoV-2 in both types of evaluated samples, regardless of the usage of PEG 8000 concentration procedure. Further, an increase in SARS-CoV-2 positivity rate was observed in samples collected in months that presented the highest number of confirmed COVID-19 cases (May/2021, June/2021 and January/2022). While PEG 8000 concentration step was found to significantly increase the positivity rate in treated wastewater samples (p < 0.005), a strong positive correlation (r: 0.84; p < 0.002) between non-concentrated raw wastewater samples with the number of new cases of COVID-19 (April/2021–February/2022) was observed. In general, the present results reinforce the efficiency of WBE approach to monitoring the presence of SARS-CoV-2 in either low- or high-capacity WWTPs. The successful usage of WBE even in raw wastewater samples makes it an interesting low-cost tool for epidemiological surveillance.

Early warning of statewide COVID-19 Omicron wave by sentineled urbanized sewer network monitoring using digital PCR in a province capital city, of Gujarat, India

Wastewater-based epidemiology (WBE) has been implemented globally. However, there remains confusion about the number and frequency of samples to be collected, as well as which types of treatment systems can provide reliable specific details about the virus prevalence in specific areas or communities, enabling prompt management and intervention measures. More research is necessary to fully comprehend the possibility of deploying sentinel locations in sewer networks in larger geographic areas. The present study introduces the first report on wastewater-based surveillance in Gandhinagar City using digital PCR (d-PCR) as a SARS-Cov-2 quantification tool, which describes the viral load from five pumping stations in Gandhinagar from October 2021 to March 2022. Raw wastewater samples (n = 119) were received and analyzed weekly to detect SARS-CoV-2 RNA, 109 of which were positive for N1 or N2 genes. The monthly variation analysis in viral genome copies depicted the highest concentrations in January 2022 and February 2022 (p < 0.05; Wilcoxon signed rank test) coincided with the Omicron wave, which contributed mainly from Vavol and Jaspur pumping stations. Cross-correlation analysis indicated that WBE from five stations in Gandhinagar, i.e., capital city sewer networks, provided two-week lead times to the citywide and statewide active cases (time-series cross-correlation function [CCF]; 0.666 and 0.648, respectively), mainly from individual contributions of the urbanized Kudasan and Vavol stations (CCF; 0.729 and 0.647, respectively). These findings suggest that sewer pumping stations in urbanized neighborhoods can be used as sentinel sites for statewide clinical surveillance and that WBE surveillance using digital PCR can be an efficient monitoring and management tool.

Assessment of rapid wastewater surveillance for determination of communicable disease spread in municipalities

Wastewater surveillance (WS) helps to improve the understanding of the spread of communicable diseases in communities. WS can assist public health decision-makers in the design and implementation of timely mitigation measures. There is an increased need to use reliable, cost-effective, simple, and rapid WS systems, given traditional analytical (or ‘gold-standard’) programs are instrument/time-intensive, and dependent on highly skilled personnel. This study investigated the application of the portable GeneXpert platform for WS of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus (IAV), influenza B virus (IBV), and respiratory syncytial virus (RSV). The GeneXpert system with the Xpert Xpress-SARS-CoV-2/Flu/RSV test kit uses reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to analyze wastewater samples. From September 2022 through January 2023, wastewater samples were collected from the influents of municipal wastewater treatment plants (MWTPs) of Saskatoon, Prince Albert, and North Battleford in the province of Saskatchewan, Canada. Both raw and concentrated wastewater samples were subjected to the GeneXpert analysis. Results showed that the Saskatoon wastewater viral loads were significantly correlated to Saskatchewan's influenza and COVID-19 clinical cases, with a lead time of 10 days for IAV and a lag time of 4 days for SARS-CoV-2. Additionally, the GeneXpert analysis of the three cities' wastewater samples showed that the raw WS could capture the dynamics of SARS-CoV-2 and IAV due to their correlation with concentrated WS. Interestingly, IBV loads were not detected in any wastewater samples, while the Saskatoon and Prince Albert wastewater samples collected following the 2023 holiday season (end of December and beginning of January) were positive for RSV. This study indicates that the GeneXpert has excellent potential for use in the development of an early warning system for transmissible disease in municipalities and limited-resource communities while simultaneously providing stakeholders with an efficient WS methodology.