Influent Samples Research Articles

Comparative Genotoxic Potential of Effluent and Influent Generated by a Beverage Industrial Plant in Lagos, Nigeria

This study was carried out to evaluate the effect of effluent and influent, from a beverage plant in Lagos state, using haematology, sperm assay, and impact of the physicochemical parameters on mice. The effluent and influent were collected in sterile bottles and were separately administered orally to 5 groups of mice at test concentrations of 6.5%, 12.5%, 25%, 50% and 100%. Mice in each group weighing approximately between 30 and 35 g were given 0.3ml/body weight of mice in each test concentration orally, daily for 21 days. Each treatment group and the control group comprised 3 mice. The resulting data showed that both test samples induced a dose-dependent, statistically significant (p<0.05) increase in the number of sperms with abnormal morphology, haematology, as well as diverse histopathological changes in the liver. The samples were also analyzed for physicochemical properties with standard laboratory techniques. The results obtained revealed that both the effluent and influent samples contain constituents (e.g. Cd, Cr and Ni) in quantities and values that are capable of inducing mutation in the biological system. The interaction of some of these constituents with the genetic material in the differentiating cells during gametogenesis may be responsible for many sperm abnormalities observed in this study. Therefore, this study is important in environmental waste management, and for the assessment of the genotoxic effects of chemicals (both solid and dissolved) in wastewater.

Municipal Wastewaters Carry Important Carbapenemase Genes Independent of Hospital Input and Can Mirror Clinical Resistance Patterns.

ABSTRACTThe spatiotemporal variation of several carbapenemase-encoding genes (CRGs) was investigated in the influent and effluent of municipal WWTPs, with or without hospital sewage input. Correlations among gene abundances, bacterial community composition, and wastewater quality parameters were tested to identify possible predictors of CRGs presence. Also, the possible role of wastewaters in mirroring clinical resistance is discussed. The taxonomic groups and gene abundances showed an even distribution among wastewater types, meaning that hospital sewage does not influence the microbial diversity and the CRG pool. The bacterial community was composed mainly of Proteobacteria, Firmicutes, Actinobacteria, Patescibacteria, and Bacteroidetes. Acinetobacter spp. was the most abundant group and had the majority of operational taxonomic units (OTUs) positively correlated with CRGs. This agrees with recent reports on clinical data. The influent samples were dominated by blaKPC, as opposed to effluent, where blaIMP was dominant. Also, blaIMP was the most frequent CRG family observed to correlate with bacterial taxa, especially with the Mycobacterium genus in effluent samples. Bacterial load, blaNDM, blaKPC, and blaOXA-48 abundances were positively correlated with BOD5, TSS, HEM, Cr, Cu, and Fe concentrations in wastewaters. When influent gene abundance values were converted into population equivalent (PE) data, the highest copies/1 PE were identified for blaKPC and blaOXA-48, agreeing with previous studies regarding clinical isolates. Both hospital and non-hospital-type samples followed a similar temporal trend of CRG incidence, but with differences among gene groups. Colder seasons favored the presence of blaNDM, blaKPC and blaOXA-48, whereas warmer temperatures show increased PE values for blaVIM and blaIMP.IMPORTANCE Wastewater-based epidemiology has recently been recognized as a valuable, cost-effective tool for antimicrobial resistance surveillance. It can help gain insights into the characteristics and distribution of antibiotic resistance elements at a local, national, and even global scale. In this study, we investigated the possible use of municipal wastewaters in the surveillance of clinically relevant carbapenemase-encoding genes (CRGs), seen as critical antibiotic resistance determinants. In this matter, our results highlight positive correlations among CRGs, microbial diversity, and wastewater physical and chemical parameters. Identified predictors can provide valuable data regarding the level of raw and treated wastewater contamination with these important antibiotic resistance genes. Also, wastewater-based gene abundances were used for the first time to observe possible spatiotemporal trends of CRGs incidence in the general population. Therefore, possible hot spots of carbapenem resistance could be easily identified at the community level, surpassing the limitations of health care-associated settings.

Influence of manganese oxide‐coated granular media on disinfection byproduct formation

AbstractThe objective of this study was to determine the influence of manganese oxide (MnOx) coated granular media on disinfection byproduct (DBP) formation for dissolved manganese (Mn(II)) and natural organic matter (NOM) concentrations typical in drinking water treatment (DWT). Instantaneous and 24‐h haloacetic acid and total trihalomethane concentrations of influent and effluent samples from two DWT plants, one using MnOx‐coated dual media (DM) filters and the other using second‐stage contactors for Mn(II) sorption by coarse MnOx‐coated ceramic media after DM filtration, were examined. Results showed that interaction with MnOx‐coated surfaces did not increase measured DBP concentrations. Observed increases in 24‐h DBP concentrations for the surface water plant were attributed to filter‐associated NOM, and not the MnOx coating. The findings of this study indicate that a second‐stage contactor – with MnOx‐coated media utilized for Mn(II) removal after a particle removal filter – does not increase DBP formation under typical treatment conditions.

Detection of SARS-CoV-2 RNA throughout wastewater treatment plants and a modeling approach to understand COVID-19 infection dynamics in Winnipeg, Canada

Although numerous studies have detected SARS-CoV-2 RNA in wastewater and attempted to find correlations between the concentration of SARS-CoV-2 RNA and the number of cases, no consensus has been reached on sample collection and processing, and data analysis. Moreover, the fate of SARS-CoV-2 in wastewater treatment plants is another issue, specifically regarding the discharge of the virus into environmental settings and the water cycle. The current study monitored SARS-CoV-2 RNA in influent and effluent wastewater samples with three different concentration methods and sludge samples over six months (July to December 2020) to compare different virus concentration methods, assess the fate of SARS-CoV-2 RNA in wastewater treatment plants, and describe the potential relationship between SARS-CoV-2 RNA concentrations in influent and infection dynamics. Skimmed milk flocculation (SMF) resulted in 15.27 ± 3.32% recovery of an internal positive control, Armored RNA, and a high positivity rate of SARS-CoV-2 RNA in stored wastewater samples compared to ultrafiltration methods employing a prefiltration step to eliminate solids in fresh wastewater samples. Our results suggested that SARS-CoV-2 RNA may predominate in solids, and therefore, concentration methods focusing on both supernatant and solid fractions may result in better recovery. SARS-CoV-2 RNA was detected in influent and primary sludge samples but not in secondary and final effluent samples, indicating a significant reduction during primary and secondary treatments. SARS-CoV-2 RNA was first detected in influent on September 30th, 2020. A decay-rate formula was applied to estimate initial concentrations of late-processed samples with SMF. A model based on shedding rate and new cases was applied to estimate SARS-CoV-2 RNA concentrations and the number of active shedders. Inferred sensitivity of observed and modeled concentrations to the fluctuations in new cases and test-positivity rates indicated a potential contribution of newly infected individuals to SARS-CoV-2 RNA loads in wastewater.

Detection and quantification of SARS-CoV-2 RNA in wastewater influent in relation to reported COVID-19 incidence in Finland

Wastewater-based surveillance is a cost-effective concept for monitoring COVID-19 pandemics at a population level. Here, SARS-CoV-2 RNA was monitored from a total of 693 wastewater (WW) influent samples from 28 wastewater treatment plants (WWTP, N = 21–42 samples per WWTP) in Finland from August 2020 to May 2021, covering WW of ca. 3.3 million inhabitants (∼ 60% of the Finnish population). Quantity of SARS-CoV-2 RNA fragments in 24 h-composite samples was determined by using the ultrafiltration method followed by nucleic acid extraction and CDC N2 RT-qPCR assay. SARS-CoV-2 RNA signals at each WWTP were compared over time to the numbers of confirmed COVID-19 cases (14-day case incidence rate) in the sewer network area.Over the 10-month surveillance period with an extensive total number of samples, the detection rate of SARS-CoV-2 RNA in WW was 79% (including 6% uncertain results, i.e., amplified only in one out of four, two original and two ten-fold diluted replicates), while only 24% of all samples exhibited gene copy numbers above the quantification limit. The range of the SARS-CoV-2 detection rate in WW varied from 33% (including 10% uncertain results) in Pietarsaari to 100% in Espoo. Only six out of 693 WW samples were positive with SARS-COV-2 RNA when the reported COVID-19 case number from the preceding 14 days was zero. Overall, the 14-day COVID-19 incidence was 7.0, 18, and 36 cases per 100 000 persons within the sewer network area when the probability to detect SARS-CoV-2 RNA in wastewater samples was 50%, 75% and 95%, respectively. The quantification of SARS-CoV-2 RNA required significantly more COVID-19 cases: the quantification rate was 50%, 75%, and 95% when the 14-day incidence was 110, 152, and 223 COVID-19 cases, respectively, per 100 000 persons. Multiple linear regression confirmed the relationship between the COVID-19 incidence and the SARS-CoV-2 RNA quantified in WW at 15 out of 28 WWTPs (overall R2 = 0.36, p < 0.001). At four of the 13 WWTPs where a significant relationship was not found, the SARS-CoV-2 RNA remained below the quantification limit during the whole study period. In the five other WWTPs, the sewer coverage was less than 80% of the total population in the area and thus the COVID-19 cases may have been inhabitants from the areas not covered.Based on the results obtained, WW-based surveillance of SARS-CoV-2 could be used as an indicator for local and national COVID-19 incidence trends. Importantly, the determination of SARS-CoV-2 RNA fragments from WW is a powerful and non-invasive public health surveillance measure, independent of possible changes in the clinical testing strategies or in the willingness of individuals to be tested for COVID-19.

Suspect and non-target screening workflow for studying the occurrence, fate, and environmental risk of contaminants in wastewater using data-independent acquisition

A comprehensive suspect and non-target screening workflow based on liquid chromatography coupled to a quadrupole-time-of-flight mass spectrometer was developed for the detection and identification of contaminants in wastewater using data-independent acquisition. The suspect screening workflow could identify 74 compounds from different classes (mainly pharmaceuticals and pesticides), of which37 compounds were confirmed by reference standards.The remaining 37 compounds were tentatively identified based on MS/MS spectra match. The occurrence and elimination of the identified compounds were studied and discussed in detail. Furthermore, the confirmed compounds were quantified where pharmaceuticals had the greatest overall concentrations in all samples, followed by flame retardants. The non-steroidal antiandrogen, bicalutamide, was detected at the highest concentration (843.9 to 3838 ng/L) at the wastewater effluents, where the flame retardant, tris(2-butoxyethyl) phosphate, exhibited a concentration in the range of 337.2 to 1304.6 ng/L. Consequently, the environmental toxicity and risk of the confirmed compounds were investigated. The pharmaceutical, telmisartan, with the insecticide, fipronil exhibited high-risk quotients ( 600∼1400 and 102∼290, respectively), demonstrating their potential toxicity at ecologically relevant amounts. Finally, multivariate analysis was applied to evaluate the efficiency of wastewater treatment . Principal component analysis was able to clearly discriminate between influent and effluent samples, demonstrating an effective treatment process..

Naive Bayes classification model for isotopologue detection in LC-HRMS data

Isotopologue identification or removal is a necessary step to reduce the number of features that need to be identified in samples analyzed with non-targeted analysis. Currently available approaches rely on either predicted isotopic patterns or an arbitrary mass tolerance, requiring information on the molecular formula or instrumental error, respectively. Therefore, a Naive Bayes isotopologue classification model was developed that does not depend on any thresholds or molecular formula information. This classification model uses the elemental mass defects of six elemental ratios and successfully identified isotopologues for both theoretical isotopic patterns and wastewater influent samples, outperforming one of the most commonly used approaches (i.e., 1.0033 ​Da mass difference method - CAMERA). For the theoretical isotopologues, the classification model outperformed an “in-house” mass difference method with a true positive rate (TPr) of 99.0% and false positive rate (FPr) of 1.8% compared to a TPr of 16.2% and an FPr of 0.02%, assuming no error. As for the wastewater influent samples, the classification model, with a TPr of 99.8% and false detection rate (FDr) of 0.5%, again performed better than the mass difference method, with a TPr of 96.3% and FDr of 4.8%. Therefore, it can be concluded that the classification model can be used for isotopologue identification, requiring no thresholds or information on the molecular formula.

Detection of SARS-CoV-2 RNA in wastewater, river water, and hospital wastewater of Nepal

The applicability of wastewater-based epidemiology (WBE) has been extensively studied throughout the world with remarkable findings. This study reports the presence and reduction of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at two wastewater treatment plants (WWTPs) of Nepal, along with river water, hospital wastewater (HWW), and wastewater from sewer lines collected between July 2020 and February 2021. SARS-CoV-2 RNA was detected in 50%, 54%, 100%, and 100% of water samples from WWTPs, river hospitals, and sewer lines, respectively, by at least one of four quantitative PCR assays tested (CDC-N1, CDC-N2, NIID_2019-nCOV_N, and N_Sarbeco). The CDC-N2 assay detected SARS-CoV-2 RNA in the highest number of raw influent samples of both WWTPs. The highest concentration was observed for an influent sample of WWTP A (5.5 ± 1.0 log10 genome copies/L) by the N_Sarbeco assay. SARS-CoV-2 was detected in 47% (16/34) of the total treated effluents of WWTPs, indicating that biological treatments installed at the tested WWTPs are not enough to eliminate SARS-CoV-2 RNA. One influent sample was positive for N501Y mutation using the mutation-specific qPCR, highlighting a need for further typing of water samples to detect Variants of Concern. Furthermore, crAssphage-normalized SARS-CoV-2 RNA concentrations in raw wastewater did not show any significant association with the number of new coronavirus disease 2019 (COVID-19) cases in the whole district where the WWTPs were located, suggesting a need for further studies focusing on suitability of viral as well as biochemical markers as a population normalizing factor. Detection of SARS-CoV-2 RNA before, after, and during the peaking in number of COVID-19 cases suggests that WBE is a useful tool for COVID-19 case estimation in developing countries.

Wastewater surveillance of SARS-CoV-2 in Austria: development, implementation, and operation of the Tyrolean wastewater monitoring program.

Wastewater-based epidemiology (WBE) is an effective approach for tracking information on spatial distribution and temporal trends of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the community level. Herein, the development, implementation, and operation of the wastewater monitoring program serving Tyrol - a federal province of Austria - are described. The development of this program was initiated by Tyrolean health authorities at the end of the first phase of the Coronavirus disease 2019 (COVID-19) pandemic (May 2020). In close co-operation with the water sector and academic institutions, efficient and effective workflows and processes for wastewater surveillance were established. The monitoring program went into operation in November 2020. By the end of July 2021, a total of 5,270 wastewater influent samples collected at 43 sites were analyzed. The monitoring program provided valuable insights into the development of the pandemic situation in Tyrol and fulfilled several tasks that are of importance in different phases of the pandemic. It represented an early-warning system, provided independent confirmation of temporal trends in COVID-19 prevalence, enabled the assessment of the effectiveness of measures, alerted about bursts of disease activity, and provided evidence for the absence of COVID-19. These findings underline the importance of establishing national wastewater monitoring programs as a complementary source of information for efficient and effective pandemic management.

Coprostanol as a Population Biomarker for SARS-CoV-2 Wastewater Surveillance Studies

Wastewater surveillance is a cost-effective tool for monitoring SARS-CoV-2 transmission in a community. However, challenges remain with regard to interpretating such studies, not least in how to compare SARS-CoV-2 levels between different-sized wastewater treatment plants. Viral faecal indicators, including crAssphage and pepper mild mottle virus, have been proposed as population biomarkers to normalise SARS-CoV-2 levels in wastewater. However, as these indicators exhibit variability between individuals and may not be excreted by everyone, their utility as population biomarkers may be limited. Coprostanol, meanwhile, is a bacterial metabolite of cholesterol which is excreted by all individuals. In this study, composite influent samples were collected from a large- and medium-sized wastewater treatment plant in Dublin, Ireland and SARS-CoV-2 N1, crAssphage, pepper mild mottle virus, HF183 and coprostanol levels were determined. SARS-CoV-2 N1 RNA was detected and quantified in all samples from both treatment plants. Regardless of treatment plant size, coprostanol levels exhibited the lowest variation in composite influent samples, while crAssphage exhibited the greatest variation. Moreover, the strongest correlations were observed between SARS-CoV-2 levels and national and Dublin COVID-19 cases when levels were normalised to coprostanol. This work demonstrates the usefulness of coprostanol as a population biomarker for wastewater surveillance studies.

Detection and Phylogenetic Analysis of Hepatitis A Virus in the Wastewater Treatment Plant of Ekbatan Town in Tehran, Iran

Background: Limited sources of fresh water necessitate the application of health policies for treatment and decontamination of human sewage for further use. A wide variety of infectious agents, including bacteria, fungi, parasites, and viruses, can be found in sewage. Enteric viruses such as hepatitis A virus (HAV) can survive the current treatments and infect susceptible hosts. Objectives: This study aimed to evaluate HAV contamination in human sewage before and after treatment in the wastewater treatment plant of Ekbatan town in Tehran, Iran, and analyze the phylogenetic properties of the identified viruses. Methods: Over a 12-month period, we collected the wastewater samples including influent, before chlorination, and effluent, from the wastewater treatment plant of Ekbatan town in Tehran, Iran. Ribonucleic acid (RNA) extraction, complementary deoxyribonucleic acid (cDNA) synthesis, and semi-nested polymerase chain reaction (PCR) were performed to identify HAV contamination. Phylogenetic analysis was performed to investigate subgenotypes of the virus. Results: HAV was detected in all influents and samples before chlorination, while the virus was detected in 50% of the effluent samples. All detected viruses belonged to subgenotype IB. Conclusions: Investigating the presence of HAV in sewage provides a general picture of the virus spread in the population of interest. HAV was detected in all influent samples, indicating that the infection is endemic in this area all year round. This also indicates the inability of the current treatment protocols in virus removal, which can be a threat to the public health.

Triclosan in Influents and Effluents from Sewage Treatment Plants Using Chlorine and UV Disinfection

Triclosan is widely used in personal care products, and has been reported to be toxic to aquatic organisms. The purpose of this study was to determine the triclosan levels in influents and effluents from three different sizes of sewage treatment plants using chlorine and ultraviolet (UV) for disinfection (i.e., STPs A, B, and C). Variations of triclosan concentration in influents and effluents (i.e., 9:00 a.m., 11:00 a.m., 13:00 p.m. and 15:00 p.m.) were also observed, respectively. The influent and effluent samples of the three STPs were composite samples collected every 2h during 9:00 a.m.-3:00 p.m by manual grab method, respectively. The samples were analyzed using a solid phase extraction (SPE) procedure, followed by liquid chromatography with tandem mass spectrometry (LC-MS-MS). Results demonstrated all of influent and effluent samples from three STPs contained triclosan. Significantly higher levels of triclosan were observed in effluent samples collected at STPs A (Median: 27.0 ng/L) as compared to in those collected at STPs B (15.0 ng/L), and C (8.0 ng/L) (p=0.012). STP A had the most serviced inhabitants and sewage flow. The triclosan levels at STP C with UV disinfection were lowest. The daytime peak of triclosan concentration in the collected influent samples was seen around 9:00 a.m. Notably, sewage discharge is the contamination source of triclosan in water bodies. Moreover, sewage flow and methods of disinfection may be the factors that affect the concentration of triclosan in effluents from sewage treatment plants.

Bio-purification of domestic wastewater through constructed wetland planted with Paspalidium flavidum.

The current study was aimed to designed laboratory scale constructed wetland (CW) for the treatment of domestic wastewater under temperature range (18-38°C) and hydraulic retention times (24 and 48 h). Besides, the soil of vegetative unit of CW was assessed using conventional culturing techniques, and 13 different bacterial species (Escherichia coli, Micrococcus, Pseudomonas, Proteus, Klebsiella, Streptococcus, Alcaligenes, Salmonella, Bacillus, Enterobacter, Staphylococcus, Shigella and Corynebacterium spp.) were determined. The pathogenic microbial load was high in influent samples, but after treatment, about 73.1-99.7% and 43.5-86.7% reduction in CFU/ml and MPN/100 ml index, respectively, were observed. Moreover, the organic loads in terms of COD, TDS, TSS, and turbidity were high in all influent samples, but after treatment, average percentage removal in different physico-chemical parameters was observed during overall treatment operations, that is, COD (59.7-65.6%), TDS (59.6-76.8%), TSS (64.9-76.7%), and turbidity (72.7-91.6%), while pH of the effluent samples was observed in the prescribed limits. It was concluded that laboratory scale CW using natural flora Paspalidium flavidum and bacterial species was efficient in the reduction of different pollution indicators and hence a best option to be modified on pilot-scale for wastewater treatment in the rural regions of Peshawar. PRACTITIONER POINTS: Role of retention times on performance of CW were studied. The commonly existing vegetation was utilized to treat domestic waste water. Both vegetations and HRT are key ingredients in obtaining high treatment efficiency.

SARS-CoV-2 RNA is enriched by orders of magnitude in primary settled solids relative to liquid wastewater at publicly owned treatment works.

Wastewater-based epidemiology has gained attention throughout the world for detection of SARS-CoV-2 RNA in wastewater to supplement clinical testing. Raw wastewater consists of small particles, or solids, suspended in liquid. Methods have been developed to measure SARS-CoV-2 RNA in the liquid and the solid fraction of wastewater, with some studies reporting higher concentrations in the solid fraction. To investigate this relationship further, six laboratories collaborated to conduct a study across five publicly owned treatment works (POTWs) where both primary settled solids obtained from primary clarifiers and raw wastewater influent samples were collected and quantified for SARS-CoV-2 RNA. Settled solids and influent samples were processed by participating laboratories using their respective methods and retrospectively paired based on date of collection. SARS-CoV-2 RNA concentrations, on a mass equivalent basis, were higher in settled solids than in influent by approximately three orders of magnitude. Concentrations in matched settled solids and influent were positively and significantly correlated at all five POTWs. RNA concentrations in both settled solids and influent were correlated to COVID-19 incidence rates in the sewersheds and thus representative of disease occurrence; the settled solids methods appeared to produce a comparable relationship between SARS-CoV-2 RNA concentration measurements and incidence rates across all POTWs. Settled solids and influent methods showed comparable sensitivity, N gene detection frequency, and calculated empirical incidence rate lower limits. Analysis of settled solids for SARS-CoV-2 RNA has the advantage of using less sample volume to achieve similar sensitivity to influent methods.

The impact of light intensity and wavelength on the performance of algal-bacterial culture treating domestic wastewater

Light is the main energy source for microalgae, and the intensity and wavelength of light influence cell metabolism and biomass composition, which, in turn, affects wastewater treatment. The objective of this study is to examine how different light intensities and light wavelengths affect the growth of mixed algal-bacterial culture while treating sewage. Three different light intensities (100, 200, and 300 μmol/m2-s) of four different light wavelengths (blue, red, white, and yellow) were selected for this study. The dissolved organic carbon (DOC), dissolved nitrogen (DN), and dissolved phosphorus (DP) in influent and effluent samples were measured, along with chlorophyll content in the biomass. The highest chlorophyll concentration of 3.5 mg/L was observed at 100 μmol/m2-s intensity of red light. The concentration of chlorophyll decreased as light intensity increased, with exception of white light. The highest DOC removal of 84% was observed at 300 μmol/m2-s intensity of blue light whereas the highest DN (51%) and DP (80%) removal was observed with a red light intensity of 100 μmol/m2-s. Overall, blue light with an intensity of 300 μmol/m2-s and red light with an intensity of 100 μmol/m2-s were found to be the most efficient at removing carbon and nutrients. The results suggested that the color and intensity of light influence algal-bacterial growth and wastewater treatment efficiency.

A method to remove cellulose from rich organic samples to analyse microplastics

Knowing the amount of microplastics that currently reach wastewater is extremely important today. Furthermore, carrying out a good quantification and detection of the type of plastic provides valuable information. However, the wastewater is loaded, in addition to a high concentration of organic matter, with a high concentration of cellulose at the treatment plant influent, which seriously hinders detection, quantification and classification of microplastics. The abundance of cellulose materials makes them possible to become false positives for microplastics. Numerous studies on the analysis of microplastics in different matrices show how to remove organic matter from samples, but there are very few studies on the removal of cellulose, which is also found in the samples and hinders their analysis. This study offers a method that combines, for the analysis of microplastics, the already known advanced oxidation treatments for the elimination of organic matter with the novel cellulose removal treatment of the samples with the aim of reducing the amount of cellulose in the influent samples of conventional wastewater treatment plants. To remove the cellulose, 40 mL of a solution of urea 8%, sodium hydroxide 8% and thiourea 6.5% (by weight) were added for every 100 mg of dry sample. The beakers were placed in the freezer at minus 20 °C for 40 min and were then placed in agitation until they reached room temperature. After that, the samples were passed through a 53 μm mesh sieve. They were washed 15 times with 30 mL of ultra-pure water. The method is called UTS because of the acronym of its reagents (Urea/Thiourea/Sodium Hydroxide). By using the UTS method it is possible to reduce almost completely the cellulose residues from the influent sewage, and sludge samples by 97.6% in a second UTS treatment and 98.2% in a third UTS treatment. In all cases analyzed, the microplastics were identified as high density polyethylene (HDPE) with correlation indices higher than 0.97, which shows that the treatment is harmless for this type of plastic material. The UTS method in combination with the WPO is an efficient and effective method for the analysis of microplastics in different matrices where cellulose and organic matter may cause possible interferences.

Removal efficiency for emerging contaminants in a WWTP from Madrid (Spain) after secondary and tertiary treatment and environmental impact on the Manzanares River

The effluents from wastewater treatment plants (WWTPs) can be an important contamination source for receiving waters. In this work, a comprehensive study on the impact of a WWTP from Madrid on the aquatic environment has been performed, including a wide number of pharmaceuticals and pesticides, among them those included in the European Watch List. 24-h composite samples of influent (IWW) and effluent wastewater after secondary (EWW2) and after secondary + tertiary treatment (EWW3) were monitored along two campaigns. Average weekly concentrations in IWW and EWW2 and EWW3 allowed estimating the removal efficiency of the WWTP for pharmaceutical active substances (PhACs). In addition, the impact of EWW3 on the water quality of the Manzanares River was assessed, in terms of PhAC and pesticide concentrations, through analysis of the river water collected upstream and downstream of the discharge point. After a preliminary risk assessment, a detailed evaluation of the impact on the aquatic environment, including a toxicological study and screening of pharmaceutical metabolites, was made for the seven most relevant PhACs: sulfamethoxazole, azithromycin and clarithromycin (antibiotics), metoprolol (antihypertensive), diclofenac (anti-inflammatory/analgesic), irbesartan (antihypertensive), and the antidepressant venlafaxine. Among selected PhACs, irbesartan, clarithromycin and venlafaxine presented moderate or high risk in the river water downstream of the discharge. Albeit no acute toxicity was detected, more detailed studies should be carried out for these substances, including additional toxicological studies, to set up potential sublethal and chronic effects on aquatic organisms.

Evaluation of multiple analytical methods for SARS-CoV-2 surveillance in wastewater samples

In this study, 14 virus concentration protocols based on centrifugation, filtration, polyethylene glycol (PEG) precipitation and ultrafiltration were tested for their efficacy for the quantification of SARS-CoV-2 in wastewater samples. These protocols were paired with four RNA extraction procedures resulting in a combination of 50 unique approaches. Bovine respiratory syncytial virus (BRSV) was used as a process control and seeded in each wastewater sample subjected to all 50 protocols. The recovery of BRSV obtained through the application of 50 unique approaches ranged from <0.03 to 64.7% (±1.6%). Combination of centrifugation as the solid removal step, ultrafiltration (Amicon-UF-15; 100 kDa cut-off; protocol 9) as the primary virus concentration method, and Zymo Quick-RNA extraction kit provided the highest BRSV recovery (64.7 ± 1.6%). To determine the impact of prolonged storage of large wastewater sample volume (900 mL) at −20 °C on enveloped virus decay, the BRSV seeded wastewaters samples were stored at −20 °C up to 110 days and analyzed using the most efficient concentration (protocol 9) and extraction (Zymo Quick-RNA kit) methods. BRSV RNA followed a first-order decay rate (k = 0.04/h with r2 = 0.99) in wastewater. Finally, 21 wastewater influent samples from five wastewater treatment plants (WWTPs) in southern Maryland, USA were analyzed between May to August 2020 to determine SARS-CoV-2 RNA concentrations. SARS-CoV-2 RNA was quantifiable in 17/21 (81%) of the influent wastewater samples with concentration ranging from 1.10 (±0.10) × 104 to 2.38 (±0.16) × 106 gene copies/L. Among the RT-qPCR assays tested, US CDC N1 assay was the most sensitive followed by US CDC N2, E_Sarbeco, and RdRp assays. Data presented in this study may enhance our understanding on the effective concentration and extraction of SARS-CoV-2 from wastewater.

Occurrence and Distribution of Antibiotic Resistance Genes in Municipal Wastewater Treatment Plants with D-Type Filters

Filters are popularly used in municipal wastewater treatment plants (WWTPs) as the final guards against effluent solids; however, their impacts on antibiotic resistance gene (ARG) removal in the WWTPs are still unclear. In this study, metagenomic analysis was used to find out the distribution characteristics of ARGs in two WWTPs equipped with the same D-Type fiber filters. Samples of influent, activated sludge liquor, secondary clarifier effluent, and D-Type filter effluent were found to host 695, 609, 675, and 643 ARG subtypes, respectively. The detected ARGs mainly included macB (4.1–8.9%), sav1866 (1.7–3.4%), and oleC (1.6–3.8%). Co-occurrence network analysis combined with contribution analysis helped to identify the ARG-related risks in the samples. Microbacterium, Acinetobacter, Gordonia, and Streptomyces significantly correlated with more than ten kinds of ARG subtypes, implying that they are potential hosts for these resistance gene subtypes. The number of ARG subtypes in the D-Type filter was less than those in the secondary clarifier effluent, indicating the potential of D-Type filters to effectively reduce the ARGs released into the environment. However, the abundance of two pathogens, Mycobacterium and PmrA, increased after the treatment by the D-Type filter, which may reveal the adverse effects of intercepting ARGs inside the fibers. The results may help the understanding of the complex role of the D-Type fiber filter on ARG distribution in WWTPs.

Bioaccumulation and health risk assessment of exposure to potentially toxic elements by consuming agricultural products irrigated with wastewater effluents

Potentially toxic elements (PTEs) have many adverse health effects due to bioaccumulation capability and their long persistence in the environment. As a renewable water source, the effluents of municipal wastewater treatment systems have been used to irrigate agricultural products widely. However, the evidence on the bioaccumulation of PTEs in crops irrigated with these effluents is still scarce, with no available study in low and middle-income countries. Therefore, this study aimed to assess the PTEs concentration in the soil and crops irrigated with effluents of Sabzevar wastewater treatment plant and the related health risk by that. The clustered method was used to determine the soil and craps samples. Seventy cumulative samples were randomly prepared in summer and autumn 2016 and 2017 from crops, soil and effluent. Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure PTEs. The health risk of exposure to PTEs was assessed using Monte Carlo simulation technique. Kruskal Wallis test and Posthoc Tukey HSD test were used to assess the mean difference of PTEs between soil, effluent and crops as well as between crops together. The bioaccumulation factor (BAF) magnitude order in different crop samples was Cd > Sr > Cu > Pb > Zn > Co > As > Cr > Ni, respectively. The Cd accumulation in Sugar beet plant was significantly higher than in other samples. The highest hazard quotient (HQ) based on single PTEs was observed for As (mineral) (mean: 5.62 × 10−1 and percentile 95th: 2.13) in Okra. Regarding total HQ (THQ), the highest and lowest mean (percentile 95th) values were 1.50 (3.22) and 2.40 × 10−1 (4.01 × 10−1) for Okra and Watermelon, respectively. The mean concentrations of Co, Cr, Ni and Zn were significantly higher in crops compared to soil and influent samples. Posthoc tests indicated that the concentration of PTEs between investigated crop samples were not statistically significant different (p > 0.05). Overall, our study suggested that irrigation with the effluent of stabilization pond wastewater treatment system exerts a potential health risk due to bioaccumulation of PTEs in crops.