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Pathogen Detection and Identification in Wastewater

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A special issue of Water (ISSN 2073-4441). This special issue belongs to the section "Wastewater Treatment and Reuse".

Deadline for manuscript submissions: closed (20 August 2023) | Viewed by 50143

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Special Issue Editors

Dr. Guangming Jiang

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Guest Editor

School of Civil, Mining and Environmental Engineering, University of Wollongong, Wollongong, NSW 2522, Australia
Interests: microbiologically influenced concrete corrosion; bio-concrete; corrosion-resistant concrete; sewer corrosion and odour management; corrosion processes; free nitrous acid; environmental microbiology; biocorrosion testing
Special Issues, Collections and Topics in MDPI journals

Dr. Ryo Honda

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Guest Editor

School of Geosciences and Civil Engineering, Kanazawa University, Kakumamachi, Kanazawa, Ishikawa 920-1192, Japan
Interests: antimicrobial resistance and SARS-CoV-2 in wastewater and water environment; membrane-applied process for wastewater treatment

Dr. Sudipti Arora

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Guest Editor

Dr. B. Lal Institute of Biotechnology, Jaipur, Rajasthan, India
Interests: wastewater treatment; wastewater based epimediology; vermifiltration; antimicrobial resistance; SARS-CoV-2 in wastewater and water environment

Special Issue Information

Dear Colleagues,

The COVID-19 pandemic has renewed research needs on the detection and monitoring of various pathogens in urban wastewater systems. Not only have the detection methods evolved from culture-based to molecular-based technology over the years, but also the purposes and applications have expanded from water quality monitoring to wastewater-based epidemiology (WBE). To ensure the effectiveness of wastewater treatment and the protection of public health, much research has been done to enhance wastewater sampling, pathogen recovery and concentration, detection sensitivity and specificity, and data analysis and modelling. These recent developments allow the more accurate and timely identification of the occurrence of pathogenic bacteria and viruses in wastewater. Meanwhile, it is still a challenge to detect new pathogens, such as SARS-CoV-2, at very low levels in wastewater. Significant research is needed to improve the detection and identifiction capability for any emerging pathogens. Genome sequencing methods are promising in providing comprehensive information including the identification of pathogens, resistance genes, and the design of PCR probes.

Public health and safety impose an ever-increasing importance on the fate and spread of pathogens in wastewater systems. To address the research needs for the detection and identification of pathogens in wastewater, we would like to invite research articles or review papers working on various aspects, including, but not limited to:

Rapid detection and identification methods
Detection of new pathogens
Development of biosensors
Sequencing-based methods
WBE studies for pathogens in wastewater
Optimisation of conventional methods
Sampling methods or passive samplers
Antimicrobial resistance in wastewater

Dr. Guangming Jiang
Dr. Ryo Honda
Dr. Sudipti Arora
Guest Editors

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Keywords

pathogen detection
wastewater-based epidemiology
sewer
wastewater treatment plant
bacteria
viruses
biosensor
PCR
DNA sequencing
antimicrobial resistance
quantitative risk assessment
variants of concern
early warning of outbreaks
transmission monitoring
wastewater surveillance

Published Papers (12 papers)

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Editorial

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4 pages, 169 KiB

Open AccessEditorial

Pathogen Detection and Identification in Wastewater

by Guangming Jiang, Ryo Honda and Sudipti Arora

Water 2024, 16(4), 611; https://0-doi-org.brum.beds.ac.uk/10.3390/w16040611 - 19 Feb 2024

Viewed by 860

Abstract

The COVID-19 pandemic has renewed research needs for the detection and monitoring of various pathogens in urban wastewater systems including sewerage systems and wastewater treatment or recycling plants [...] Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

Research

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15 pages, 1098 KiB

Open AccessCommunication

The Reduction of SARS-CoV-2 RNA Concentration in the Presence of Sewer Biofilms

by Shuxin Zhang, Elipsha Sharma, Ananda Tiwari, Yan Chen, Samendra P. Sherchan, Shuhong Gao, Xu Zhou, Jiahua Shi and Guangming Jiang

Water 2023, 15(11), 2132; https://0-doi-org.brum.beds.ac.uk/10.3390/w15112132 - 04 Jun 2023

Cited by 2 | Viewed by 1891

Abstract

Wastewater surveillance has been widely used to track the prevalence of SARS-CoV-2 in communities. Although some studies have investigated the decay of SARS-CoV-2 RNA in wastewater, understanding about its fate during wastewater transport in real sewers is still limited. This study aims to assess the impact of sewer biofilms on the dynamics of SARS-CoV-2 RNA concentration in naturally contaminated real wastewater (raw influent wastewater without extra SARS-CoV-2 virus/gene seeding) using a simulated laboratory-scale sewer system. The results indicated that, with the sewer biofilms, a 90% concentration reduction of the SARS-CoV-2 RNA was observed within 2 h both in wastewater of gravity (GS, gravity-driven sewers) and rising main (RM, pressurized sewers) sewer reactors. In contrast, the 90% reduction time was 8–26 h in control reactors without biofilms. The concentration reduction of SARS-CoV-2 RNA in wastewater was significantly more in the presence of sewer biofilms. In addition, an accumulation of c.a. 260 and 110 genome copies/cm² of the SARS-CoV-2 E gene was observed in the sewer biofilm samples from RM and GS reactors within 12 h, respectively. These results confirmed that the in-sewer concentration reduction of SARS-CoV-2 RNA in wastewater was likely caused by the partition to sewer biofilms. The need to investigate the in-sewer dynamic of SARS-CoV-2 RNA, such as the variation of RNA concentration in influent wastewater caused by biofilm attachment and detachment, was highlighted by the significantly enhanced reduction rate of SARS-CoV-2 RNA in wastewater of sewer biofilm reactors and the accumulation of SARS-CoV-2 RNA in sewer biofilms. Further research should be conducted to investigate the in-sewer transportation of SARS-CoV-2 and their RNA and evaluate the role of sewer biofilms in leading to underestimates of COVID-19 prevalence in communities. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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12 pages, 2776 KiB

Open AccessArticle

Molecular Identification of Human Adenovirus Isolated from Different Wastewater Treatment Plants in Riyadh, Saudi Arabia: Surveillance and Meteorological Impacts

by Khalid Maniah, Islam Nour, Atif Hanif, Mohamed Taha Yassin, Abdulrahman Alkathiri, Ibrahim Al-Ashkar and Saleh Eifan

Water 2023, 15(7), 1367; https://0-doi-org.brum.beds.ac.uk/10.3390/w15071367 - 02 Apr 2023

Cited by 2 | Viewed by 1985

Abstract

Regular water environment monitoring is crucial for minimizing contamination caused by waterborne viruses and reducing health risks. As the human adenovirus (HAdV) is linked to clinical episodes of gastroenteritis in children, the present investigation aimed to detect HAdVs in three wastewater treatment plants in Riyadh, Saudi Arabia (King Saud University (KSU-WWTP), Manfoha (MN-WWTP), and Embassy Quarter (EMB-WWTP)). The impact of seasonal variability and meteorological factors on the prevalence of HAdVs was also investigated. The HAdV hexon sequences of the isolated human adenoviruses were phylogenetically analyzed and revealed that the F species of HAdV, especially serotype 41, dominated. The highest prevalence of HAdV was detected in KSU-WWTP (83.3%), followed by MN-WWTP (75%), and EMB-WWTP (66.6%). Seasonal distribution insignificantly influenced the HAdV prevalence among sampling areas (p > 0.05). The highest prevalence of HAdVs (100%) was detected in late Summer and Autumn at temperatures (high: 34–43 °C, low: 18–32 °C) and moderate prevalence of 66.67% in Winter (particularly, in January and February) at lower temperature ranges (high: 26 °C, low: 10 °C–12 °C). The large variation of HAdV prevalence detected at different humidity ranges emphasized the significant impact of relative humidity on HAdV incidence in raw water of WWTPs (p = 0.009, R² = 0.419). In contrast, wind speed was detected to have insignificant influence on HAdV prevalence among different WWTPs (p > 0.05, R² = 0.03). The study provides important data for the incidence of HAdVs in wastewater treatments plants in Riyadh, Saudi Arabia, which enabled the successful management of health hazards of viral diseases transmitted via fecal-oral route. In addition, the non-significant influence of seasonal variability on HAdV prevalence highlights the potentiality of utilizing HAdVs as a potential fecal indicator of wastewater contamination. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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10 pages, 1517 KiB

Open AccessCommunication

MIMS as a Low-Impact Tool to Identify Pathogens in Water

by Salvia Sajid, Ishika Aryal, Suleman Farooq Chaudhri, Frants Roager Lauritsen, Mikkel Girke Jørgensen, Håvard Jenssen and Bala Krishna Prabhala

Water 2023, 15(1), 184; https://0-doi-org.brum.beds.ac.uk/10.3390/w15010184 - 02 Jan 2023

Cited by 2 | Viewed by 2483

Abstract

Bacteria produce many kinds of volatile compounds throughout their lifecycle. Identifying these volatile compounds can help to understand bacterial interactions with the host and/or other surrounding pathogens of the same or different species. Some commonly used techniques to detect these volatile compounds are GC and/or LC coupled to mass spectrometric techniques. However, these methods can sometimes become challenging owing to tedious sample preparation steps. Thus, identifying an easier method to detect these volatile compounds was investigated in the present study. Here, Membrane-inlet mass spectrometry (MIMS) provided a facile low-impact alternative to the existing strategies. MIMS was able to differentiate between the pathogenic and nonpathogenic bacterial strains, implying that it can be used as a bioprocess monitoring tool to analyze water samples from either water treatment plants or biotechnological industries. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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12 pages, 1862 KiB

Open AccessArticle

Extensive Wastewater-Based Epidemiology as a Resourceful Tool for SARS-CoV-2 Surveillance in a Low-to-Middle-Income Country through a Successful Collaborative Quest: WBE, Mobility, and Clinical Tests

by Juan Eduardo Sosa-Hernández, Mariel Araceli Oyervides-Muñoz, Elda M. Melchor-Martínez, Erin M. Driver, Devin A. Bowes, Simona Kraberger, Sofia Liliana Lucero-Saucedo, Rafaela S. Fontenele, Lizeth Parra-Arroyo, LaRinda A. Holland, Samantha Ayde Peña-Benavides, Melanie Engstrom Newell, Manuel Martínez-Ruiz, Sangeet Adhikari, Laura Isabel Rodas-Zuluaga, Rahul Kumar, Itzel Y. López-Pacheco, Carlos Castillo-Zacarias, Hafiz M. N. Iqbal, Efrem S. Lim, Daniel Salas-Limón, Arvind Varsani, Rolf U. Halden and Roberto Parra-Saldívar Show full author list Hide full author list

Water 2022, 14(12), 1842; https://0-doi-org.brum.beds.ac.uk/10.3390/w14121842 - 08 Jun 2022

Cited by 9 | Viewed by 3121

Abstract

The COVID-19 pandemic has challenged healthcare systems worldwide. Efforts in low-to-middle-income countries (LMICs) cannot keep stride with infection rates, especially during peaks. A strong international collaboration between Arizona State University (ASU), Tec de Monterrey (TEC), and Servicios de Agua y Drenaje de Monterrey (Local Water Utilities) is acting to integrate wastewater-based epidemiology (WBE) of SARS-CoV-2 in the region as a complementary approach to aid the healthcare system. Wastewater was collected from four sewer catchments in the Monterrey Metropolitan area in Mexico (pop. 4,643,232) from mid-April 2020 to February 2021 (44 weeks, n = 644). Raw wastewater was filtered and filter-concentrated, the RNA was extracted using columns, and the Charité/Berlin protocol was used for the RT-qPCR. The viral loads obtained between the first (June 2020) and second waves (February 2021) of the pandemic were similar; in contrast, the clinical cases were fewer during the first wave, indicating poor coverage. During the second wave of the pandemic, the SARS-CoV-2 quantification in wastewater increased 14 days earlier than the COVID-19 clinical cases reported. This is the first long-term WBE study in Mexico and demonstrates its value in pandemic management. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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12 pages, 618 KiB

Open AccessArticle

Evaluation of Pre-Analytical and Analytical Methods for Detecting SARS-CoV-2 in Municipal Wastewater Samples in Northern Italy

by Laura Pellegrinelli, Sara Castiglioni, Clementina E. Cocuzza, Barbara Bertasi, Valeria Primache, Silvia Schiarea, Giulia Salmoiraghi, Andrea Franzetti, Rosario Musumeci, Michela Tilola, Elisa Galuppini, Giorgio Bertanza, Marialuisa Callegari, Fabrizio Stefani, Andrea Turolla, Emanuela Ammoni, Danilo Cereda, Elena Pariani, Sandro Binda and the WBE Study Group

Water 2022, 14(5), 833; https://0-doi-org.brum.beds.ac.uk/10.3390/w14050833 - 07 Mar 2022

Cited by 9 | Viewed by 2509

Abstract

(1) Background: The surveillance of SARS-CoV-2 RNA in urban wastewaters allows one to monitor the presence of the virus in a population, including asymptomatic and symptomatic individuals, capturing the real circulation of this pathogen. The aim of this study was to evaluate the performance of different pre-analytical and analytical methods for identifying the presence of SARS-CoV-2 in untreated municipal wastewaters samples by conducting an inter-laboratory proficiency test. (2) Methods: three methods of concentration, namely, (A) Dextran and PEG-6000 two-phase separation, (B) PEG-8000 precipitation without a chloroform purification step and (C) PEG-8000 precipitation with a chloroform purification step were combined with three different protocols of RNA extraction by using commercial kits and were tested by using two primers/probe sets in three different master mixes. (3) Results: PEG-8000 precipitation without chloroform treatment showed the best performance in the SARS-CoV-2 recovery; no major differences were observed among the protocol of RNA extraction and the one-step real-time RT-PCR master mix kits. The highest analytic sensitivity was observed by using primers/probe sets targeting the N1/N3 fragments of SARS-CoV-2. (4) Conclusions: PEG-8000 precipitation in combination with real-time RT-PCR targeting the N gene (two fragments) was the best performing workflow for the detection of SARS-CoV-2 RNA in municipal wastewaters. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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19 pages, 4809 KiB

Open AccessArticle

Monitoring of SARS-CoV-2 Variants by Wastewater-Based Surveillance as a Sustainable and Pragmatic Approach—A Case Study of Jaipur (India)

by Aditi Nag, Sudipti Arora, Vikky Sinha, Ekta Meena, Devanshi Sutaria, Akhilendra Bhushan Gupta and Krishna Mohan Medicherla

Water 2022, 14(3), 297; https://0-doi-org.brum.beds.ac.uk/10.3390/w14030297 - 19 Jan 2022

Cited by 15 | Viewed by 3176

Abstract

Wastewater-based surveillance has been emerging as an efficient and advantageous tool to predict COVID-19 prevalence in the population, much earlier (7–28 days) than reported clinical cases, thus providing sufficient time to organize resources and optimize their use in managing COVID-19. Since the commencement of the COVID-19 pandemic, SARS-CoV-2 genetic lineages have emerged and are circulating all over the world. The assessment of SARS-CoV-2 variants of concern (VOCs) in wastewater has recently been proven to be successful. The present research demonstrates a case study utilizing an established approach to perform monitoring of SARS-CoV-2 variants from 11 distinct wastewater treatment plants across Jaipur (India) during the second peak period of COVID-19 (from 19 February 2021 to 8 June 2021). The sequences obtained were analyzed to detect lineage using the Pangolin tool and SNPs using the mpileup utility of Samtools, which reported high genome coverage. The mutation analyses successfully identified the penetration of the B.1. in the first two weeks of sampling (19–26 February), followed by the B.1.617.2 variant into Jaipur in the first week of March 2021. B.1.617.2 was initially discovered in India in October 2020; however, it was not reported until early April 2021.The present study identified the presence of B.1.617.2 in early March, which correlates well with the clinical patient’s data (290 cases were reported much later by the government on 10 May 2021). The average total genome coverage of the samples is 94.39% when mapped onto the severe acute respiratory syndrome coronavirus 2 isolate Wuhan-Hu-1; a complete genome (NC_045512.2) sequence and SNP analysis showed that 37–51 SNPs were identified in each sample. The current study demonstrates that sewage surveillance for variant characterization is a reliable and practical method for tracking the diversity of SARS-CoV-2 strains in the community that is considerably faster than clinical genomic surveillance. As a result, this method can predict the advent of epidemiologically or clinically important mutations/variants, which can help with public health decision making. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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16 pages, 2436 KiB

Open AccessFeature PaperArticle

Imprints of Lockdown and Treatment Processes on the Wastewater Surveillance of SARS-CoV-2: A Curious Case of Fourteen Plants in Northern India

by Sudipti Arora, Aditi Nag, Ankur Rajpal, Vinay Kumar Tyagi, Satya Brat Tiwari, Jasmine Sethi, Devanshi Sutaria, Jayana Rajvanshi, Sonika Saxena, Sandeep Kumar Shrivastava, Vaibhav Srivastava, Akhilendra Bhushan Gupta, Absar Ahmed Kazmi and Manish Kumar

Water 2021, 13(16), 2265; https://0-doi-org.brum.beds.ac.uk/10.3390/w13162265 - 19 Aug 2021

Cited by 14 | Viewed by 3328

Abstract

The present study investigated the detection of severe acute respiratory syndrome–coronavirus 2 (SARS-CoV-2) genomes at each treatment stage of 14 aerobic wastewater treatment plants (WWTPs) serving the major municipalities in two states of Rajasthan and Uttarakhand in Northern India. The untreated, primary, secondary and tertiary treated wastewater samples were collected over a time frame ranging from under-lockdown to post-lockdown conditions. The results showed that SARS-CoV-2 RNA was detected in 13 out of 40 wastewater samples in Jaipur district, Rajasthan and in 5 out of 14 wastewater samples in the Haridwar District, Uttarakhand with the E gene predominantly observed as compared to the N and RdRp target genes in later time-points of sampling. The Ct values of genes present in wastewater samples were correlated with the incidence of patient and community cases of COVID-19. This study further indicates that the viral RNA could be detected after the primary treatment but was not present in secondary or tertiary treated samples. This study implies that aerobic biological wastewater treatment systems such as moving bed biofilm reactor (MBBR) technology and sequencing batch reactor (SBR) are effective in virus removal from the wastewater. This work might present a new indication that there is little to no risk in relation to SARS-CoV-2 while reusing the treated wastewater for non-potable applications. In contrast, untreated wastewater might present a potential route of viral transmission through WWTPs to sanitation workers and the public. However, there is a need to investigate the survival and infection rates of SARS-CoV-2 in wastewater. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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16 pages, 956 KiB

Open AccessArticle

Comparison of Detecting and Quantitating SARS-CoV-2 in Wastewater Using Moderate-Speed Centrifuged Solids versus an Ultrafiltration Method

by Maria Hasing, Jiaao Yu, Yuanyuan Qiu, Rasha Maal-Bared, Sudha Bhavanam, Bonita Lee, Steve Hrudey and Xiaoli Pang

Water 2021, 13(16), 2166; https://0-doi-org.brum.beds.ac.uk/10.3390/w13162166 - 06 Aug 2021

Cited by 10 | Viewed by 2600

Abstract

Mounting evidence suggests that solids are a reliable matrix for SARS-CoV-2 detection in wastewater, yet studies comparing solids-based methods and common concentration methods using the liquid fraction remain limited. In this study, we developed and optimized a method for SARS-CoV-2 detection in wastewater using moderate-speed centrifuged solids and evaluated it against an ultrafiltration reference method. SARS-CoV-2 was quantified in samples from 12 wastewater treatment plants from Alberta, Canada, using RT-qPCR targeting the N2 and E genes. PCR inhibition was examined by spiking salmon DNA. The effects of using different amounts of solids, adjusting the sample pH to 9.6–10, and modifying the elution volume at the final step of RNA extraction were evaluated. SARS-CoV-2 detection rate in solids from 20 mL of wastewater showed no statistically significant difference compared to the ultrafiltration method (97/139 versus 90/139, p = 0.26, McNemar’s mid-p test). The optimized wastewater solids-based method had a significantly lower rate of samples with PCR inhibition versus ultrafiltration (3% versus 9.5%, p = 0.014, Chi-square test). Our optimized moderate-speed centrifuged solids-based method had similar sensitivity when compared to the ultrafiltration reference method but had the added advantages of lower costs, fewer processing steps, and a shorter turnaround time. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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Review

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14 pages, 722 KiB

Open AccessReview

Effects of Temperature and Water Types on the Decay of Coronavirus: A Review

by Ying Guo, Yanchen Liu, Shuhong Gao, Xu Zhou, Muttucumaru Sivakumar and Guangming Jiang

Water 2023, 15(6), 1051; https://0-doi-org.brum.beds.ac.uk/10.3390/w15061051 - 09 Mar 2023

Cited by 4 | Viewed by 2272

Abstract

The analysis of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) gene copy numbers in wastewater samples can provide quantitative information on Coronavirus Disease-19 (COVID-19) cases within a sewer catchment. However, many wastewater-based epidemiology (WBE) studies have neglected virus decay during the wastewater transportation process in sewers while back-calculating COVID-19 prevalence. Among various sewer condition parameters, wastewater temperature and dilution by fresh/saltwater infiltration may result in a significant change to the virus decay, in terms of both infectivity and Ribonucleic Acid (RNA). This paper reviewed the literature to identify and discuss the effects of temperature and water types (i.e., wastewater, freshwater, and seawater) on coronavirus decay based on the decay rate constants that were collected from published papers. To evaluate the importance of virus decay, a sensitivity analysis was then conducted with decay rates of SARS-CoV-2 RNA based on a WBE back-calculation equation. Finally, the decay rates of coronavirus in wastewater were also compared with those of other viruses to further understand the difference among virus species. The decay of SARS-CoV-2 RNA was found to be less impacted by temperature variation than viable coronaviruses. Nevertheless, WBE back-calculation was still sensitive to the RNA decay rates increased by warm wastewater (i.e., over 26 °C), which could lead to a two-times higher relative variance in estimated COVID-19 prevalence, considering the wastewater temperature variation between 4 and 37 °C in a sewer catchment with a 12-h hydraulic retention time. Comparatively, the sensitivity of the WBE estimation to the enveloped SARS-CoV-2 was greater than nonenveloped enteric viruses, which were less easily degradable in wastewater. In addition, wastewater dilution by stormwater inflow and accompanied cold weather might alleviate the decay of coronavirus infectivity, thus increasing the potential risk of COVID-19 transmission through wastewater. Overall, this paper aims to better understand the impact of in-sewer processes on coronavirus decay and its potential implications for WBE. The outcome could quantitatively inform WBE and improve awareness of the increased risk of COVID-19 infection via wastewater during heavy rainfall events. Given the identified scarcity of data available for coronavirus decay in salt water or with chemical additions, future research on the fate of SARS-CoV-2 subjected to chemical dosing for sewer or wastewater treatment plant operations is recommended. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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19 pages, 1223 KiB

Open AccessEditor’s ChoiceReview

Tracing COVID-19 Trails in Wastewater: A Systematic Review of SARS-CoV-2 Surveillance with Viral Variants

by Ananda Tiwari, Sangeet Adhikari, Shuxin Zhang, Tamunobelema B. Solomon, Anssi Lipponen, Md. Aminul Islam, Ocean Thakali, Sarawut Sangkham, Mohamed N. F. Shaheen, Guangming Jiang, Eiji Haramoto, Payal Mazumder, Bikash Malla, Manish Kumar, Tarja Pitkänen and Samendra P. Sherchan

Water 2023, 15(6), 1018; https://0-doi-org.brum.beds.ac.uk/10.3390/w15061018 - 07 Mar 2023

Cited by 11 | Viewed by 9217

Abstract

The emergence of new variants of SARS-CoV-2 associated with varying infectivity, pathogenicity, diagnosis, and effectiveness against treatments challenged the overall management of the COVID-19 pandemic. Wastewater surveillance (WWS), i.e., monitoring COVID-19 infections in communities through detecting viruses in wastewater, was applied to track the emergence and spread of SARS-CoV-2 variants globally. However, there is a lack of comprehensive understanding of the use and effectiveness of WWS for new SARS-CoV-2 variants. Here we systematically reviewed published articles reporting monitoring of different SARS-CoV-2 variants in wastewater by following the PRISMA guidelines and provided the current state of the art of this study area. A total of 80 WWS studies were found that reported different monitoring variants of SARS-CoV-2 until November 2022. Most of these studies (66 out of the total 80, 82.5%) were conducted in Europe and North America, i.e., resource-rich countries. There was a high variation in WWS sampling strategy around the world, with composite sampling (50/66 total studies, 76%) as the primary method in resource-rich countries. In contrast, grab sampling was more common (8/14 total studies, 57%) in resource-limited countries. Among detection methods, the reverse transcriptase polymerase chain reaction (RT-PCR)-based sequencing method and quantitative RT-PCR method were commonly used for monitoring SARS-CoV-2 variants in wastewater. Among different variants, the B1.1.7 (Alpha) variant that appeared earlier in the pandemic was the most reported (48/80 total studies), followed by B.1.617.2 (Delta), B.1.351 (Beta), P.1 (Gamma), and others in wastewater. All variants reported in WWS studies followed the same pattern as the clinical reporting within the same timeline, demonstrating that WWS tracked all variants in a timely way when the variants emerged. Thus, wastewater monitoring may be utilized to identify the presence or absence of SARS-CoV-2 and follow the development and transmission of existing and emerging variants. Routine wastewater monitoring is a powerful infectious disease surveillance tool when implemented globally. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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31 pages, 492 KiB

Open AccessFeature PaperReview

Molecular Methods for Pathogenic Bacteria Detection and Recent Advances in Wastewater Analysis

by Shuxin Zhang, Xuan Li, Jiangping Wu, Lachlan Coin, Jake O’Brien, Faisal Hai and Guangming Jiang

Water 2021, 13(24), 3551; https://0-doi-org.brum.beds.ac.uk/10.3390/w13243551 - 12 Dec 2021

Cited by 17 | Viewed by 13379

Abstract

With increasing concerns about public health and the development of molecular techniques, new detection tools and the combination of existing approaches have increased the abilities of pathogenic bacteria monitoring by exploring new biomarkers, increasing the sensitivity and accuracy of detection, quantification, and analyzing various genes such as functional genes and antimicrobial resistance genes (ARG). Molecular methods are gradually emerging as the most popular detection approach for pathogens, in addition to the conventional culture-based plate enumeration methods. The analysis of pathogens in wastewater and the back-estimation of infections in the community, also known as wastewater-based epidemiology (WBE), is an emerging methodology and has a great potential to supplement current surveillance systems for the monitoring of infectious diseases and the early warning of outbreaks. However, as a complex matrix, wastewater largely challenges the analytical performance of molecular methods. This review synthesized the literature of typical pathogenic bacteria in wastewater, types of biomarkers, molecular methods for bacterial analysis, and their recent advances in wastewater analysis. The advantages and limitation of these molecular methods were evaluated, and their prospects in WBE were discussed to provide insight for future development. Full article

(This article belongs to the Special Issue Pathogen Detection and Identification in Wastewater)

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