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1.
Arch Microbiol ; 204(8): 513, 2022 Jul 22.
Article in English | MEDLINE | ID: covidwho-1941517

ABSTRACT

The plant pathogen pepper mild mottle virus (PMMoV) has recently been proposed as a water quality indicator, it is a RNA virus belonging to the genus Tobamovirus in the family Virgoviridae that causes harm to the pepper crops. After consuming processed food products containing infected peppers, such as hot sauces, PMMoV is excreted in high concentrations in feces; therefore, this is the most common RNA virus, constantly found in the feces of humans. The fecal-oral pathway is emerging as an environmental problem. The presence of high concentrations of pathogens associated with human excreta in environmental waters or water reuse supplies poses a threat to public health. Due to the difficulty in determining the presence of pathogens effectively in water, attempts to monitor microbial water quality often use surrogates or indicator organisms that can be easily detected; therefore, PMMoV is used as a viral surrogate in aquatic environment. This paper describes the incidence and persistence of PMMoV in aquatic environments and in waste treatment plants and its usefulness for quantifying virus reductions by advanced water treatment technologies. In recent research, SARS-CoV-2 was reported to be found in wastewater and utilized for the purpose of monitoring coronavirus illness outbreaks. Since PMMoV is readily identified in the human feces and can also serve as an indicator of human waste, the determined PMMoV concentrations may be utilized to give the normalized report of the SARS-CoV-2 concentration, so that, the amount of human waste found in the wastewater can be taken into consideration.


Subject(s)
COVID-19 , Tobamovirus , Feces , Humans , SARS-CoV-2 , Tobamovirus/genetics , Waste Water , Water Microbiology
2.
Int J Environ Res Public Health ; 19(13)2022 06 30.
Article in English | MEDLINE | ID: covidwho-1934060

ABSTRACT

INTRODUCTION: There is growing interest in the public health and transport sectors in research into exposure to biological hazards, considering not only the risks arising from inter-human contagion, but also those related to exposure to the flight environment itself. The aim of this paper is to report data from an investigation into the water and air-conditioning systems of commercial aircraft for the presence of Legionella contamination, with a total of 645 water samples taken during the period 2007-2021. METHODS: The investigation involved 126 aircraft of six different commercial aircraft types: MD80, Airbus A320 F, Embraer 175/190, AIRBUS A330, Boeing 767 and Boeing 777. Water samples were taken from the water systems (toilet taps, galley and boilers). Each sample was preliminarily subjected to an evaluation of the following parameters: temperature, pH and residual chlorine. The ScanVit® Legionella kit was used for bacteria detection and enumeration. RESULTS: Samples were considered positive if the number of colony-forming units/liter (CFU/L) was >100. For the entire observation period, 45% of the investigated aircraft tested positive. Regarding the overall number of samples analyzed, 68.4% (441/645) were below 100 CFU/L, and thus within the limits allowed by the Italian Guidelines. CONCLUSIONS: Water system contamination with Legionella in the air transport field is a real public health issue that should not be underestimated given the heavy passenger traffic. Infection should be considered an occupational risk to which crew members are exposed.


Subject(s)
Legionella pneumophila , Legionella , Chlorine/analysis , Humans , Water , Water Microbiology , Water Pollution
3.
Nat Commun ; 13(1): 635, 2022 02 03.
Article in English | MEDLINE | ID: covidwho-1671556

ABSTRACT

Tracking SARS-CoV-2 genetic diversity is strongly indicated because diversifying selection may lead to the emergence of novel variants resistant to naturally acquired or vaccine-induced immunity. To monitor New York City (NYC) for the presence of novel variants, we deep sequence most of the receptor binding domain coding sequence of the S protein of SARS-CoV-2 isolated from the New York City wastewater. Here we report detecting increasing frequencies of novel cryptic SARS-CoV-2 lineages not recognized in GISAID's EpiCoV database. These lineages contain mutations that had been rarely observed in clinical samples, including Q493K, Q498Y, E484A, and T572N and share many mutations with the Omicron variant of concern. Some of these mutations expand the tropism of SARS-CoV-2 pseudoviruses by allowing infection of cells expressing the human, mouse, or rat ACE2 receptor. Finally, pseudoviruses containing the spike amino acid sequence of these lineages were resistant to different classes of receptor binding domain neutralizing monoclonal antibodies. We offer several hypotheses for the anomalous presence of these lineages, including the possibility that these lineages are derived from unsampled human COVID-19 infections or that they indicate the presence of a non-human animal reservoir.


Subject(s)
SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Waste Water/virology , Water Microbiology , Adult , Aged , Animals , Antibodies, Monoclonal/immunology , Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , COVID-19/virology , Female , Genetic Variation , High-Throughput Nucleotide Sequencing , Humans , Male , Mice , Middle Aged , Mutation , New York City , Protein Binding , Rats , Spike Glycoprotein, Coronavirus/immunology , Young Adult
4.
Viruses ; 14(1)2022 01 13.
Article in English | MEDLINE | ID: covidwho-1625824

ABSTRACT

Infection with enterovirus D68 (EV-D68) has been linked with severe neurological disease such as acute flaccid myelitis (AFM) in recent years. However, active surveillance for EV-D68 is lacking, which makes full assessment of this association difficult. Although a high number of EV-D68 infections were expected in 2020 based on the EV-D68's known biannual circulation patterns, no apparent increase in EV-D68 detections or AFM cases was observed during 2020. We describe an upsurge of EV-D68 detections in wastewater samples from the United Kingdom between July and November 2021 mirroring the recently reported rise in EV-D68 detections in clinical samples from various European countries. We provide the first publicly available 2021 EV-D68 sequences showing co-circulation of EV-D68 strains from genetic clade D and sub-clade B3 as in previous years. Our results show the value of environmental surveillance (ES) for the early detection of circulating and clinically relevant human viruses. The use of a next-generation sequencing (NGS) approach helped us to estimate the prevalence of EV-D68 viruses among EV strains from other EV serotypes and to detect EV-D68 minor variants. The utility of ES at reducing gaps in virus surveillance for EV-D68 and the possible impact of nonpharmaceutical interventions introduced to control the COVID-19 pandemic on EV-D68 transmission dynamics are discussed.


Subject(s)
Enterovirus D, Human/isolation & purification , Waste Water/virology , COVID-19/epidemiology , COVID-19/prevention & control , Capsid Proteins/genetics , Enterovirus D, Human/classification , Enterovirus D, Human/genetics , Humans , Phylogeny , RNA, Viral/genetics , SARS-CoV-2 , Sequence Analysis, DNA , United Kingdom/epidemiology , Wastewater-Based Epidemiological Monitoring , Water Microbiology
5.
J Water Health ; 19(6): 918-932, 2021 Dec.
Article in English | MEDLINE | ID: covidwho-1613432

ABSTRACT

Human noroviruses are a leading cause of food- and water-borne disease, which has led to an interest in quantifying norovirus health risks using quantitative microbial risk assessment (QMRA). Given the limited availability of quantitative norovirus data to input to QMRA models, some studies have applied a conversion factor to estimate norovirus exposure based on measured fecal indicator bacteria (FIB) concentrations. We conducted a review of peer-reviewed publications to identify the concentrations of noroviruses and FIB in raw, secondary-treated, and disinfected wastewater. A meta-analysis was performed to determine the ratios of norovirus-FIB pairs in each wastewater matrix and the variables that significantly impact these ratios. Norovirus-to-FIB ratios were found to be significantly impacted by the norovirus genotype, month of sample collection, geographic location, and the extent of wastewater treatment. Additionally, we evaluated the impact of using a FIB-to-virus conversion factor in QMRA and found that the choice of conversion ratio has a great impact on estimated health risks. For example, the use of a conversion ratio previously used in the World Health Organization Guidelines for the Safe Use of Wastewater, Excreta and Greywater predicted health risks that were significantly lower than those estimated with measured norovirus concentrations used as inputs. This work emphasizes the gold standard of using measured pathogen concentrations directly as inputs to exposure assessment in QMRA. While not encouraged, if one must use a FIB-to-virus conversion ratio to estimate norovirus dose, the ratio should be chosen carefully based on the target microorganisms (i.e., strain, genotype, or class), prevalence of disease, and extent of wastewater treatment.


Subject(s)
Norovirus , Waste Water , Bacteria , Feces , Humans , Risk Assessment , Water Microbiology
6.
Viruses ; 13(11)2021 10 26.
Article in English | MEDLINE | ID: covidwho-1526862

ABSTRACT

Despite a surge of RNA virome sequencing in recent years, there are still many RNA viruses to uncover-as indicated by the relevance of viral dark matter to RNA virome studies (i.e., putative viruses that do not match to taxonomically identified viruses). This study explores a unique site, a high-rate algal pond (HRAP), for culturing industrially microalgae, to elucidate new RNA viruses. The importance of viral-host interactions in aquatic systems are well documented, and the ever-expanding microalgae industry is no exception. As the industry becomes a more important source of sustainable plastic manufacturing, a producer of cosmetic pigments and alternative protein sources, and a means of CO2 remediation in the face of climate change, studying microalgal viruses becomes a vital practice for proactive management of microalgae cultures at the industrial level. This study provides evidence of RNA microalgal viruses persisting in a CO2 remediation pilot project HRAP and uncovers the diversity of the RNA virosphere contained within it. Evidence shows that family Marnaviridae is cultured in the basin, alongside other potential microalgal infecting viruses (e.g., family Narnaviridae, family Totitiviridae, and family Yueviridae). Finally, we demonstrate that the RNA viral diversity of the HRAP is temporally dynamic across two successive culturing seasons.


Subject(s)
Microalgae/virology , Phylogeny , Ponds , RNA Viruses/classification , Water Microbiology , Animals , Biodiversity , Biomass , Metagenome , Pilot Projects , RNA Viruses/genetics , Rotifera/virology , Seasons , Water
7.
Sci Total Environ ; 806(Pt 2): 150616, 2022 Feb 01.
Article in English | MEDLINE | ID: covidwho-1440355

ABSTRACT

Stagnant water can cause water quality deterioration and, in particular, microbiological contaminations, posing potential health risks to occupants. University buildings were unoccupied with little water usage during the COVID-19 pandemic. It's an opportunity to study microbiological quality of long-term stagnant water (LTSW) in university buildings. The tap water samples were collected for three months from four types of campus buildings to monitor water quality and microbial risks after long-term stagnation. Specifically, the residual chlorine, turbidity, and iron/zinc were disqualified, and the heterotrophic plate counts (HPC) exceeded the Chinese national standard above 100 times. It took 4-54 days for these parameters to recover to the routine levels. Six species of pathogens were detected with high frequency and levels (101-105 copies/100 mL). Remarkably, L. pneumophilia occurred in 91% of samples with turbidity > 1 NTU. The absence of the culturable cells for these bacteria possibly implied their occurrence in a viable but non-culturable (VBNC) status. The bacterial community of the stagnant tap water differed significantly and reached a steady state in more than 50 days. Furthermore, a high concentration of endotoxin (>10 EU/mL) was found in LTSW, which was in accordance with the high proportion of dead bacteria. The results suggested that the increased microbiological risks require more attention and the countermeasures before the building reopens should be taken.


Subject(s)
COVID-19 , Water Supply , Humans , Pandemics , SARS-CoV-2 , Universities , Water Microbiology , Water Quality
8.
Am J Infect Control ; 49(12): 1564-1566, 2021 12.
Article in English | MEDLINE | ID: covidwho-1415167

ABSTRACT

Prolonged building closures are prevalent during the COVID-19 pandemic, resulting in extreme stagnation in building water systems. High-throughput sequencing analysis revealed significantly increased presence of Legionella due to extreme water stagnation, highlighting elevated exposure risks to Legionella from building water systems during re-opening of previously closed buildings.


Subject(s)
COVID-19 , Drinking Water , Legionella , Humans , Pandemics , SARS-CoV-2 , Water Microbiology , Water Supply
10.
Viruses ; 13(9)2021 09 10.
Article in English | MEDLINE | ID: covidwho-1411079

ABSTRACT

We used wastewater-based epidemiology and amplicon-based long-read high-throughput sequencing for surveillance of enteroviruses (EVs) in Maricopa County, Arizona, Southwest United States. We collected 48 samples from 13 sites in three municipalities between 18 June and 1 October 2020, and filtered (175 mL each; 0.45 µm pore size) and extracted RNA from the filter-trapped solids. The RNA was converted to cDNA and processed through two workflows (Sanger sequencing (SSW) and long-read Illumina sequencing (LRISW)) each including a nested polymerase chain reaction (nPCR) assay. We subjected the ~350 bp amplicon from SSW to Sanger sequencing and the ~1900-2400 bp amplicon from LRISW to Illumina sequencing. We identified EV contigs from 11 of the 13 sites and 41.67% (20/48) of screened samples. Using the LRISW, we detected nine EV genotypes from three species (Enterovirus A (CVA4, EV-A76, EV-A90), Enterovirus B (E14) and Enterovirus C (CVA1, CVA11, CVA13, CVA19 and CVA24)) with Enterovirus C representing approximately 90% of the variants. However, the SSW only detected the five Enterovirus C types. Similarity and phylogenetic analysis showed that multiple Enterovirus C lineages were circulating, co-infecting and recombining in the population during the season despite the SARS-CoV-2 pandemic and the non-pharmaceutical public health measures taken to curb transmission.


Subject(s)
Enterovirus Infections/epidemiology , Enterovirus Infections/virology , Enterovirus/genetics , Waste Water/microbiology , Water Microbiology , Arizona/epidemiology , Enterovirus/isolation & purification , Enterovirus Infections/history , High-Throughput Nucleotide Sequencing , History, 21st Century , Humans , Phylogeny , RNA, Viral , Seasons , Wastewater-Based Epidemiological Monitoring
11.
Int J Environ Res Public Health ; 18(17)2021 09 06.
Article in English | MEDLINE | ID: covidwho-1403598

ABSTRACT

"Ensure access to water for all", states Goal 6 of the UN's Sustainable Development Goals. This worldwide challenge requires identifying the best water disinfection method for each scenario. Traditional methods have limitations, which include low effectiveness towards certain pathogens and the formation of disinfection byproducts. Solar-driven methods, such as solar water disinfection (SODIS) or solar photocatalysis, are novel, effective, and financially and environmentally sustainable alternatives. We have conducted a critical study of publications in the field of water disinfection using solar energy and, hereby, present the first bibliometric analysis of scientific literature from Elsevier's Scopus database within the last 20 years. Results show that in this area of growing interest USA, Spain, and China are the most productive countries in terms of publishing, yet Europe hosts the most highly recognized research groups, i.e., Spain, Switzerland, Ireland, and UK. We have also reviewed the journals in which researchers mostly publish and, using a systematic approach to determine the actual research trends and gaps, we have analyzed the capacity of these publications to answer key research questions, pinpointing six clusters of keywords in relation to the main research challenges, open areas, and new applications that lie ahead. Most publications focused on SODIS and photocatalytic nanomaterials, while a limited number focused on ensuring adequate water disinfection levels, testing regulated microbial indicators and emerging pathogens, and real-world applications, which include complex matrices, large scale processes, and exhaustive cost evaluation.


Subject(s)
Disinfection , Water Purification , Sunlight , Water , Water Microbiology
12.
Int J Hyg Environ Health ; 237: 113836, 2021 08.
Article in English | MEDLINE | ID: covidwho-1392331

ABSTRACT

Our surrounding environment has been influenced by the COVID-19 pandemic situation. The second wave of COVID-19 in India has proven to be more devastating and aggressive than the first wave of the pandemic, which led to recognizing India as one of the world's topmost worst-hit nations considering >4000 fatalities reported in a single day in May 2021. Such "resurgence and acceleration" of COVID-19 transmission has been fuelled by the MahaKumbh festival and political mass gathering (elections rallies) events, where the COVID-19 protocols have been ignored by millions of pilgrims/followers. The present review discusses only the consequences of this year's MahaKumbh festivals, the largest religious mass gathering on earth, which was held during the COVID-19 pandemic in India, and its impact on both the spread of SARS-CoV-2 among participants and their families and its influence on the quality of the river Ganga. This article tries to give readers outside of India an overview of how much impact of any such single large gathering of any relgion in any part of the world can drive coronavirus infections and effectively commence the second/third wave outbreak with this case study. Furthermore, the religious large scale celebration are widely accepted through out the world that have played a significant role in the spread of the pandemic into remote villages and towns all over the subcontinent/world, thus affecting many areas with insufficient healthcare facilities that have been relatively spared. This review also highlights the potential risk of transmission from infected humans into the aquatic environment of the river Ganga. Besides the obvious relevance of SARS-CoV-2, a large variety of other water-related disease vectors (bacteria, viruses, and protozoa) stemming from visitors to the religious congregation were introduced into the upstream regions of the Ganga river. Their sheer number is assumed to have had a severe influence on its delicate ecosystem, including endangered mammals such as the river Dolphins. The detailed epidemiological and clinical study on transmission routes of SARS-CoV-2 is the need of the hour to understand the pathogenesis of RNA virus infection and prevent the massive spreading of such infectious respiratory diseases. An interdisciplinary approach, rooted in evidence-based efficient learning, contextual strategies, and a streamlined unified approach should be adopted to help in the development of a proactive prevention model during future MahaKumbh festival (and similar religious gatherings) instead of just "picking up the pieces" in a conventional post-event model.


Subject(s)
COVID-19/transmission , Holidays , Rivers/microbiology , SARS-CoV-2 , Animals , Endangered Species , Humans , India , Water Microbiology , Water Pollution
13.
Microb Biotechnol ; 13(6): 1689-1701, 2020 11.
Article in English | MEDLINE | ID: covidwho-1343797

ABSTRACT

The COVID-19 pandemic is not only a challenge for public health and hospitals, but affects many aspects of our societies. This Lilliput minireview deals with problems that the pandemic causes for the food industry, addressing the presence and persistence of SARS-CoV-2 in the food environment, methods of virus inactivation and the protection of the food worker and the consumer. So far food has not been implicated in the transmission of the infection, but social disruptions caused by the pandemic could cause problems with food security.


Subject(s)
Betacoronavirus , Coronavirus Infections/transmission , Food Industry , Food Supply , Pneumonia, Viral/transmission , Animals , Betacoronavirus/isolation & purification , COVID-19 , Coronavirus Infections/prevention & control , Disease Models, Animal , Feces/virology , Humans , Occupational Exposure/prevention & control , Pandemics/prevention & control , Pneumonia, Viral/prevention & control , SARS-CoV-2 , Sewage/virology , Water Microbiology
14.
Salud Publica Mex ; 63(1, ene-feb): 109-119, 2020 Dec 22.
Article in Spanish | MEDLINE | ID: covidwho-1310298

ABSTRACT

Objetivo. Describir la evidencia sobre la presencia e infectividad de SARS-CoV-2 y otros coronavirus en aguas residuales y su potencial uso como herramienta de vigilancia epidemiológica. Material y métodos. Búsqueda de publicaciones en PubMed y medRxiv desde enero 2003 hasta el 8 de junio de 2020 de acuerdo con la guía de revisiones rápidas de Cochrane. Resultados. Se incluyeron 29 publicaciones. El ARN de SARS-CoV-2 no infectivo se encontró en agua residual hospitalaria, agua residual cruda, tratada y lodos de plantas de tratamiento. Los niveles cuantitativos de ARN viral en agua residual presentan relación con el número de casos de Covid-19. SARS-CoV-1 y otros coronavirus permanecieron infectivos en agua residual cruda hasta por dos días. Conclusiones. Hasta esta revisión no existe evidencia sobre la presencia de virus infectivos de SARS-CoV-2 en agua residual cruda o tratada. La cuantificación de ARN de SARS-CoV-2 en agua residual es útil para la vigilancia epidemiológica.


Subject(s)
RNA, Viral/isolation & purification , SARS-CoV-2/isolation & purification , Waste Water/virology , Wastewater-Based Epidemiological Monitoring , Coronavirus/isolation & purification , Coronavirus/pathogenicity , Mexico , SARS Virus/isolation & purification , SARS Virus/pathogenicity , SARS-CoV-2/genetics , SARS-CoV-2/pathogenicity , Virulence , Water Microbiology
15.
PLoS One ; 16(7): e0254540, 2021.
Article in English | MEDLINE | ID: covidwho-1309963

ABSTRACT

Coronaviruses (CoVs) are a family of viruses that are best known as the causative agents of human diseases like the common cold, Middle East Respiratory Syndrome (MERS), Severe Acute Respiratory Syndrome (SARS) and COVID-19. CoVs spread by human-to-human transmission via droplets or direct contact. There is, however, concern about potential waterborne transmission of SARS-CoV-2, the virus responsible for COVID-19, as it has been found in wastewater facilities and rivers. To date, little is known about the stability of SARS-CoV-2 or any other free coronavirus in aquatic environments. The inactivation of terrestrial CoVs in seawater is rarely studied. Here, we use a porcine respiratory coronavirus (PRCV) that is commonly found in animal husbandry as a surrogate to study the stability of CoVs in natural water. A series of experiments were conducted in which PRCV (strain 91V44) was added to filtered and unfiltered fresh- and saltwater taken from the river Scheldt and the North Sea. Virus titres were then measured by TCID50-assays using swine testicle cell cultures after various incubation times. The results show that viral inactivation of PRCV in filtered seawater can be rapid, with an observed 99% decline in the viral load after just two days, which may depend on temperature and the total suspended matter concentration. PRCV degraded much slower in filtered water from the river Scheldt, taking over 15 days to decline by 99%, which was somewhat faster than the PBS control treatment (T99 = 19.2 days). Overall, the results suggest that terrestrial CoVs are not likely to accumulate in marine environments. Studies into potential interactions with exudates (proteases, nucleases) from the microbial food web are, however, recommended.


Subject(s)
Coronavirus Infections/transmission , Porcine Respiratory Coronavirus/isolation & purification , Testis/cytology , Waste Water/virology , Animals , Cells, Cultured , Filtration , Male , Pilot Projects , Porcine Respiratory Coronavirus/pathogenicity , Rivers/virology , Swine , Testis/virology , Time Factors , Viral Load , Water Microbiology
16.
Chemosphere ; 281: 130728, 2021 Oct.
Article in English | MEDLINE | ID: covidwho-1233383

ABSTRACT

As the coronavirus disease 2019 continues to spread globally, its culprit, the severe acute respiratory syndrome coronavirus 2 has been brought under scrutiny. In addition to inhalation transmission, the possible fecal-oral viral transmission via water/wastewater has also been brought under the spotlight, necessitating a timely global review on the current knowledge about waterborne viruses in drinking water treatment system - the very barrier that intercepts waterborne pathogens to terminal water users. In this article we reviewed the occurrence, concentration methods, and control strategies, also, treatment performance on waterborne viruses during drinking water treatment were summarized. Additionally, we emphasized the potential of applying the quantitative microbial risk assessment to guide drinking water treatment to mitigate the viral exposure risks, especially when the unregulated novel viral pathogens are of concern. This review paves road for better control of viruses at drinking water treatment plants to protect public health.


Subject(s)
COVID-19 , Drinking Water , Viruses , Water Purification , Humans , SARS-CoV-2 , Water Microbiology
18.
Rev Environ Health ; 36(3): 309-317, 2021 Sep 27.
Article in English | MEDLINE | ID: covidwho-1088789

ABSTRACT

An important group of viruses are Coronaviruses that affect the health of people worldwide, in particular the acute respiratory syndrome. The present work has addressed the updated literature on the topic of coronaviruses transmission through water and wastewater as well as identified gaps in research to inform future studies. In total, 198 articles were selected, then after screening, 48 eligible studies were fully reviewed. Accordingly, the studies showed that the coronavirus has been isolated and identified from water as well as wastewater. The results of researches show that the presence of SARS-Co-2 virus in municipal wastewater is possible due to the excretion of the virus in human feces. In addition, the SARS-Co-2 virus was isolated from contaminated water and rivers, but there is insufficient evidence for virus transmission by water and wastewater. Water and wastewater treatment methods are able to reduce the pollution load caused by this virus in water sources. Water disinfection has an effective role in removing it from water and wastewater sources. Due to the short period of time in the global pandemic and the small number of studies in this field, further studies are needed to make a definite statement about the transferability of virus in water and wastewater.


Subject(s)
COVID-19/transmission , SARS-CoV-2/isolation & purification , Waste Water/virology , Water Microbiology , Humans , Pandemics , Water Pollution/analysis , Water Purification/standards
20.
Int J Hyg Environ Health ; 233: 113692, 2021 04.
Article in English | MEDLINE | ID: covidwho-1056688

ABSTRACT

The aim of the present study was to develop a simple, sensitive, and specific approach to quantifying the SARS-CoV-2 genome in wastewater and to evaluate this approach as a means of epidemiological surveillance. Twelve wastewater samples were collected from a metropolitan area in north-eastern France during April and May 2020. In addition to the quantification of the SARS-CoV-2 genome, F-specific RNA phages of genogroup II (FRNAPH GGII), naturally present in wastewater, were used as an internal process control for the viral concentration and processing of RT-PCR inhibitors. A concentration method was required to allow the quantification of the SARS-CoV-2 genome over the longest possible period. A procedure combining ultrafiltration, phenol-chloroform-isoamyl alcohol purification, and the additional purification of the RNA extracts was chosen for the quantification of the SARS-CoV-2 genome in 100-mL wastewater samples. At the same time, the COVID-19 outbreak was evaluated through patients from the neighbouring University Hospital of Nancy, France. A regular decrease in the concentration of the SARS-CoV-2 genome from ~104 gc/L to ~102 gc/L of wastewater was observed over the eight weeks of the study, during which the population was placed under lockdown. The SARS-CoV-2 genome was even undetectable during one week in the second half of May and present but non-quantifiable in the last sample (28 May). A concordant circulation in the human community was highlighted by virological diagnosis using respiratory samples, which showed a decrease in the number of COVID-19 cases from 677 to 52 per week over the same period. The environmental surveillance of COVID-19 using a reliable viral quantification procedure to test wastewater is a key approach. The real-time detection of viral genomes can allow us to predict and monitor the circulation of SARS-CoV-2 in clinical settings and survey the entire urban human population.


Subject(s)
COVID-19/epidemiology , Disease Outbreaks , Environmental Monitoring/methods , Genome, Viral , SARS-CoV-2/genetics , Waste Water/microbiology , COVID-19/diagnosis , COVID-19/virology , Chemical Precipitation , Cities/epidemiology , France/epidemiology , Hospitals, University , Humans , Ultrafiltration , Viral Proteins/chemistry , Viral Proteins/isolation & purification , Water Microbiology
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