Wastewater surveillance for viral pathogens: A tool for public health.

A focus on water quality has intensified globally, considering its critical role in sustaining life and ecosystems. Wastewater, reflecting societal development, profoundly impacts public health. Wastewater-based epidemiology (WBE) has emerged as a surveillance tool for detecting outbreaks early, monitoring infectious disease trends, and providing real-time insights, particularly in vulnerable communities. WBE aids in tracking pathogens, including viruses, in sewage, offering a comprehensive understanding of community health and lifestyle habits. With the rise in global COVID-19 cases, WBE has gained prominence, aiding in monitoring SARS-CoV-2 levels worldwide. Despite advancements in water treatment, poorly treated wastewater discharge remains a threat, amplifying the spread of water-, sanitation-, and hygiene (WaSH)-related diseases. WBE, serving as complementary surveillance, is pivotal for monitoring community-level viral infections. However, there is untapped potential for WBE to expand its role in public health surveillance. This review emphasizes the importance of WBE in understanding the link between viral surveillance in wastewater and public health, highlighting the need for its further integration into public health management.

Illicit drugs in Brazil: environmental consequences and consumption patterns.

This article reviews the literature on the consumption, street drug analysis, distribution, and main environmental impacts of illicit drugs in Brazil and analyzes the III National Survey on Drug Use by the Brazilian Population. The literature review is based on articles published in national and international journals between 2018 and 2023. This review consists of two analyses, the first of which addresses publications from the last 6 years on the monitoring of illicit drugs in Brazil and a second analysis based on the III National Survey on Drug Use that addresses the different possibilities of contact with drugs. The results revealed that the Southeast region of Brazil has the highest number of studies on the subject, especially in the state of São Paulo, while the North and Northeast regions have the lowest number of studies. The Midwest regions only have studies in the federal capital city, Brasília, while no studies were found in states bordering countries that produce illicit drugs, such as Paraguay and Bolivia. Analytical methods that use the concept of miniaturization, green chemistry, and the adoption of acceptance methods are frequent in most articles. Chemometric and statistical tools are widely used for the analysis, development, and conclusion of identification and quantification methods. Among the articles studied, there was a predominance in the analysis of cocaine metabolites and cannabis metabolites in the aquatic environment, where their concentrations ranged from 0.01 to 2000 ng L-1. Studies also reported bioaccumulation in marine biota with concentrations of up to 4.58 µg kg-1 for mussels and sediments, posing a risk to algae, crustaceans, and fish. Furthermore, the data show that the consumption of illicit drugs is increasing in Brazil, especially among young people.

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

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

Detection of SARS-CoV-2 in Wastewater Associated with Scientific Stations in Antarctica and Possible Risk for Wildlife.

Before December 2020, Antarctica had remained free of COVID-19 cases. The main concern during the pandemic was the limited health facilities available at Antarctic stations to deal with the disease as well as the potential impact of SARS-CoV-2 on Antarctic wildlife through reverse zoonosis. In December 2020, 60 cases emerged in Chilean Antarctic stations, disrupting the summer campaign with ongoing isolation needs. The SARS-CoV-2 RNA was detected in the wastewater of several scientific stations. In Antarctica, treated wastewater is discharged directly into the seawater. No studies currently address the recovery of infectious virus particles from treated wastewater, but their presence raises the risk of infecting wildlife and initiating new replication cycles. This study highlights the initial virus detection in wastewater from Antarctic stations, identifying viral RNA via RT-qPCR targeting various genomic regions. The virus's RNA was found in effluent from two wastewater plants at Maxwell Bay and O'Higgins Station on King George Island and the Antarctic Peninsula, respectively. This study explores the potential for the reverse zoonotic transmission of SARS-CoV-2 from humans to Antarctic wildlife due to the direct release of viral particles into seawater. The implications of such transmission underscore the need for continued vigilance and research.

Identifying outbreaks in sewer networks: An adaptive sampling scheme under network's uncertainty.

Motivated by the implementation of a SARS-Cov-2 sewer surveillance system in Chile during the COVID-19 pandemic, we propose a set of mathematical and algorithmic tools that aim to identify the location of an outbreak under uncertainty in the network structure. Given an upper bound on the number of samples we can take on any given day, our framework allows us to detect an unknown infected node by adaptively sampling different network nodes on different days. Crucially, despite the uncertainty of the network, the method allows univocal detection of the infected node, albeit at an extra cost in time. This framework relies on a specific and well-chosen strategy that defines new nodes to test sequentially, with a heuristic that balances the granularity of the information obtained from the samples. We extensively tested our model in real and synthetic networks, showing that the uncertainty of the underlying graph only incurs a limited increase in the number of iterations, indicating that the methodology is applicable in practice.

Wastewater surveillance to assess cocaine and methylenedioxymethamphetamine use trends during a major music festival in Brazil.

Wastewater-based epidemiology was applied in northeastern Brazil during a dance festival, revealing that cocaine consumption doubled during the event days. The daily drug loads were 0.95 ± 0.03 to 11.4 ± 0.4 g/day for BE, 1.8 ± 0.4 to 7.6 ± 0.3 g/day for COC, 0.04 ± 0.02 to 0.19 ± 0.02 g/day for COE, and 0.08 ± 0.02 to 0.80 ± 0.02 g/day for MDMA.

This study employed the wastewater­based epidemiology (WBE) strategy to investigate cocaine (COC) and ecstasy (MDMA) usage during Carnatal festival in Rio Grande do Norte, Brazil. Wastewater samples were collected over the four festival days (December 12­15, 2019), as well as during corresponding days before and after the event. The samples underwent filtration, acidification, solid­phase extraction (SPE) for pre­concentration, and analysis using ultra­high­performance liquid chromatography­tandem mass spectrometry (UHPLC­MS/MS) with calibration using deuterated internal standards. The measured concentration ranges in the wastewater samples were as follows: benzoylecgonine (BE, a metabolite of COC use) ranged from 760 ± 30 to 8914 ± 252 ng/L, COC (non­metabolized or directly discharged) ranged from 1182 ± 25 to 6,760 ± 95 ng/L, cocaethylene (COE, a metabolite of alcohol­COC use) ranged from 21 ± 10 to 161 ± 10 ng/L, and MDMA (ecstasy) ranged from 18 ± 5 to 613 ± 15 ng/L. Daily wastewater loads were estimated as 0.81 ± 0.03 to 11.69 ± 0.33 g/day for BE, 1.42 ± 0.03 to 8.06 ± 0.11 g/day for COC, 0.02 ± 0.01 to 0.21 ± 0.01 g/day for COE, and 0.02 ± 0.01 to 0.80 ± 0.02 g/day for MDMA. Notably, COE and MDMA were only detected above the quantification limit during the festival days. The data enable estimating that COC consumption doubled during the event days. The COC/BE ratio indicated likely drug disposals on the event's last day and the subsequent week. The study's findings shed light on illicit substance consumption, assisting in public health programs and combating drug trafficking efforts.

Cross-continental comparative experiences of wastewater surveillance and a vision for the 21st century.

The COVID-19 pandemic has brought the epidemiological value of monitoring wastewater into sharp focus. The challenges of implementing and optimising wastewater monitoring vary significantly from one region to another, often due to the array of different wastewater systems around the globe, as well as the availability of resources to undertake the required analyses (e.g. laboratory infrastructure and expertise). Here we reflect on the local and shared challenges of implementing a SARS-CoV-2 monitoring programme in two geographically and socio-economically distinct regions, São Paulo state (Brazil) and Wales (UK), focusing on design, laboratory methods and data analysis, and identifying potential guiding principles for wastewater surveillance fit for the 21st century. Our results highlight the historical nature of region-specific challenges to the implementation of wastewater surveillance, including previous experience of using wastewater surveillance, stakeholders involved, and nature of wastewater infrastructure. Building on those challenges, we then highlight what an ideal programme would look like if restrictions such as resource were not a constraint. Finally, we demonstrate the value of bringing multidisciplinary skills and international networks together for effective wastewater surveillance.

Urine biomarkers for Alzheimer's disease: A new opportunity for wastewater-based epidemiology?

While Alzheimer's disease (AD) diagnosis, management, and care have become priorities for healthcare providers and researcher's worldwide due to rapid population aging, epidemiologic surveillance efforts are currently limited by costly, invasive diagnostic procedures, particularly in low to middle income countries (LMIC). In recent years, wastewater-based epidemiology (WBE) has emerged as a promising tool for public health assessment through detection and quantification of specific biomarkers in wastewater, but applications for non-infectious diseases such as AD remain limited. This early review seeks to summarize AD-related biomarkers and urine and other peripheral biofluids and discuss their potential integration to WBE platforms to guide the first prospective efforts in the field. Promising results have been reported in clinical settings, indicating the potential of amyloid ß, tau, neural thread protein, long non-coding RNAs, oxidative stress markers and other dysregulated metabolites for AD diagnosis, but questions regarding their concentration and stability in wastewater and the correlation between clinical levels and sewage circulation must be addressed in future studies before comprehensive WBE systems can be developed.

Detection and diversity of gastrointestinal viruses in wastewater from Caracas, Venezuela, 2021-2022.

Gastrointestinal viruses (GIV) are an important cause of childhood morbidity and mortality, particularly in developing countries. Their epidemiological impact in Venezuela during the COVID-19 pandemic remains unclear. GIV can also be detected in domestic sewage. Ninety-one wastewater samples from urban areas of Caracas collected over 12 months and concentrated by polyethylene-glycol-precipitation, were analyzed by multiplex reverse-transcription-PCR for rotavirus/calicivirus/astrovirus and enterovirus/klassevirus/cosavirus, and monoplex-PCR for adenovirus and Aichi virus. The overall frequency of virus detection was 46.2%, fluctuating over months, and peaking in the rainy season. Adenoviruses circulated throughout the year, especially type F41, and predominated (52.7%) over caliciviruses (29.1%) that peaked in the rainy months, rotaviruses (9.1%), cosaviruses (5.5%), astroviruses and enteroviruses (1.8%). Aichi-virus and klassevirus were absent. Rotavirus G9/G12, and P[4]/P[8]/P[14] predominated. The occurrence of GIV in wastewater reflects transmission within the population of Caracas and the persistence of a potential public health risk that needs to be adequately monitored.

Monkeypox Virus in Wastewater Samples from Santiago Metropolitan Region, Chile.

Sewage surveillance provides useful epidemiologic and public health information on viral infections at the population level. We detected monkeypox virus DNA from sewage samples covering 85% of the population in Santiago Metropolitan Region Chile. We also isolated infective viruses from those samples. Wastewater surveillance could complement clinical surveillance for monkeypox virus.

Risk assessment of a Brazilian urban population due to the exposure to pyrethroid insecticides during the COVID-19 pandemic using wastewater-based epidemiology.

Pyrethroids are synthetic insecticides commonly used in agriculture and homes due to their low toxicity to mammals and effectiveness at low doses. However, exposure to pyrethroids can cause various symptoms, depending on the route of exposure. To measure human exposure to pyrethroids, researchers used wastewater-based epidemiology (WBE) with polar organic chemical integrative samplers (POCIS) sampling. This approach is a cost-effective and efficient way to assess exposure to pyrethroids. The study aimed to evaluate the exposure of an urban population in Brazil to pyrethroids during the COVID-19 pandemic using WBE with POCIS sampling. Researchers analyzed 3-phenoxybenzoic acid (3-PBA) in wastewater using passive sampling with POCIS, which was extracted with methanol and analyzed using UPLC-MS/MS. The range of CTWA concentrations of 3-PBA in wastewater was 24.3-298.2 ng L-1, with a mean value of 134 ± 76.5 ng L-1. The values were used to estimate the exposure of the population to pyrethroid insecticides. Three different conversion factors were applied to determine the range of exposure to at least 20 different pyrethroid insecticides. The exposure values ranged from 18.08 to 1441.49 mg day-1 per 1000 inhabitants. The toxicological risk posed to the exposed population was evaluated by calculating the WBE toxicological level (WBE-TL). Lambda-cyhalothrin was used as a reference for risk assessment, and the WBE-TL values for lambda-cyhalothrin ranged from 0.5 to 8.29 (considering the high CF). We compared mobility trends to 3-PBA exposure during the COVID-19 pandemic. The study highlighted the effectiveness of POCIS sampling in WBE and provided useful information for policymakers and regulatory agencies. POCIS sampling has practical advantages, including analyte pre-concentration, low operational cost, and ease of use. Overall, the study shows the importance of monitoring and understanding the exposure of the population to pyrethroid insecticides, especially during the pandemic when people may be spending more time at home.

Surveillance of SARS-CoV-2, rotavirus, norovirus genogroup II, and human adenovirus in wastewater as an epidemiological tool to anticipate outbreaks of COVID-19 and acute gastroenteritis in a city without a wastewater treatment plant in the Peruvian Highlands.

The coronavirus disease 2019 (COVID-19) pandemic has demonstrated that Wastewater Based Epidemiology is a fast and economical alternative for monitoring severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the community level in high-income countries. In the present study, wastewater from a city in the Peruvian Highlands, which lacks a wastewater treatment plant, was monitored for one year to assess the relationship between the concentration of SARS-CoV-2 and the reported cases of COVID-19 in the community. Additionally, we compared the relationship between rotavirus (RV), norovirus genogroup II (NoV GGII), and human adenovirus (HAdV) with the number of reported cases of acute gastroenteritis. Before commencing the analysis of the samples, the viral recovery efficacy of three processing methods was determined in spiked wastewater with SARS-CoV-2. This evaluation demonstrated the highest recovery rate with direct analysis (72.2 %), as compared to ultrafiltration (50.8 %) and skimmed milk flocculation (5.6 %). Wastewater monitoring revealed that 72 % (36/50) of the samples tested positive for SARS-CoV-2, with direct analysis yielding the highest detection frequency and quantification of SARS-CoV-2. Furthermore, a strong correlation was observed between the concentration of SARS-CoV-2 in wastewater and the reported cases of COVID-19, mainly when we shift the concentration of SARS-CoV-2 by two weeks, which allows us to anticipate the onset of the fourth and fifth waves of the pandemic in Peru up to two weeks in advance. All samples processed using the skimmed milk flocculation method tested positive and showed high concentrations of RV, NoV GGII, and HAdV. In fact, the highest RV concentrations were detected up to four weeks before outbreaks of acute gastroenteritis reported in children under four years of age. In conclusion, the results of this study suggest that periodic wastewater monitoring is an excellent epidemiological tool for surveillance and can anticipate outbreaks of infectious diseases, such as COVID-19, in low- and middle-income countries.

Zika virus RNA persistence and recovery in water and wastewater: An approach for Zika virus surveillance in resource-constrained settings.

During the 2015-2016 Zika virus (ZIKV) epidemic in the Americas, serological cross-reactivity with other flaviviruses and relatively high costs of nucleic acid testing in the region hindered the capacity for widespread diagnostic testing. In such cases where individual testing is not feasible, wastewater monitoring approaches may offer a means of community-level public health surveillance. To inform such approaches, we characterized the persistence and recovery of ZIKV RNA in experiments where we spiked cultured ZIKV into surface water, wastewater, and a combination of both to examine the potential for detection in open sewers serving communities most affected by the ZIKV outbreak, such as those in Salvador, Bahia, Brazil. We used reverse transcription droplet digital PCR to quantify ZIKV RNA. In our persistence experiments, we found that the persistence of ZIKV RNA decreased with increasing temperature, significantly decreased in surface water versus wastewater, and significantly decreased when the initial concentration of virus was lowered by one order of magnitude. In our recovery experiments, we found higher percent recovery of ZIKV RNA in pellets versus supernatants from the same sample, higher recoveries in pellets using skimmed milk flocculation, lower recoveries of ZIKV RNA in surface water versus wastewater, and lower recoveries from a freeze thaw. We also analyzed samples collected from Salvador, Brazil during the ZIKV outbreak (2015-2016) that consisted of archived samples obtained from open sewers or environmental waters thought to be contaminated by sewage. Although we did not detect any ZIKV RNA in the archived Brazil samples, results from these persistence and recovery experiments serve to inform future wastewater monitoring efforts in open sewers, an understudied and important application of wastewater monitoring.

Evaluation of low-cost SARS-CoV-2 RNA purification methods for viral quantification by RT-qPCR and next-generation sequencing analysis: Implications for wider wastewater-based epidemiology adoption.

Based Epidemiology (WBE) consists of quantifying biomarkers in sewerage systems to derive real-time information on the health and/or lifestyle of the contributing population. WBE usefulness was vastly demonstrated in the context of the COVID-19 pandemic. Many methods for SARS-CoV-2 RNA determination in wastewater were devised, which vary in cost, infrastructure requirements and sensitivity. For most developing countries, implementing WBE for viral outbreaks, such as that of SARS-CoV-2, proved challenging due to budget, reagent availability and infrastructure constraints. In this study, we assessed low-cost methods for SARS-CoV-2 RNA quantification by RT-qPCR, and performed variant identification by NGS in wastewater samples. Results showed that the effect of adjusting pH to 4 and/or adding MgCl2 (25 mM) was negligible when using the adsorption-elution method, as well as basal physicochemical parameters in the sample. In addition, results supported the standardized use of linear rather than plasmid DNA for a more accurate viral RT-qPCR estimation. The modified TRIzol-based purification method in this study yielded comparable RT-qPCR estimation to a column-based approach, but provided better NGS results, suggesting that column-based purification for viral analysis should be revised. Overall, this work provides evaluation of a robust, sensitive and cost-effective method for SARS-CoV-2 RNA analysis that could be implemented for other viruses, for a wider WEB adoption.

20-Month monitoring of SARS-CoV-2 in wastewater of Curitiba, in Southern Brazil.

The COVID-19 pandemic resulted in the collapse of healthcare systems and led to the development and application of several approaches of wastewater-based epidemiology to monitor infected populations. The main objective of this study was to carry out a SARS-CoV-2 wastewater based surveillance in Curitiba, Southern Brazil Sewage samples were collected weekly for 20 months at the entrance of five treatment plants representing the entire city and quantified by qPCR using the N1 marker. The viral loads were correlated with epidemiological data. The correlation by sampling points showed that the relationship between the viral loads and the number of reported cases was best described by a cross-correlation function, indicating a lag between 7 and 14 days amidst the variables, whereas the data for the entire city presented a higher correlation (0.84) with the number of positive tests at lag 0 (sampling day). The results also suggest that the Omicron VOC resulted in higher titers than the Delta VOC. Overall, our results showed that the approach used was robust as an early warning system, even with the use of different epidemiological indicators or changes in the virus variants in circulation. Therefore, it can contribute to public decision-makers and health interventions, especially in vulnerable and low-income regions with limited clinical testing capacity. Looking toward the future, this approach will contribute to a new look at environmental sanitation and should even induce an increase in sewage coverage rates in emerging countries.

Changing molecular epidemiology of Hepatitis A virus in Santiago, Chile from 2010 to 2021.

OBJECTIVES: Hepatitis A (HAV) virus causes asymptomatic to life-treating fulminant hepatitis. During infection, patients show large viral excretion in their stools. Resistance of HAV to environmental conditions, allows us to recover viral nucleotide sequences from wastewater and trace its evolutionary history. METHODS: We characterize twelve years of HAV circulation in wastewater from Santiago, Chile, and conducted phylogenetic analyses to decipher the dynamics of circulating lineages. RESULTS: We observed the exclusive circulation of the HAV IA genotype. The molecular epidemiologic analyses showed a steady circulation of a dominant lineage with low genetic diversity (d = 0,007) between 2010 and 2017. An outbreak of Hepatitis A associated with men who have sex with men, in 2017 was associated with the irruption of a new lineage. Remarkably, a dramatic change in the dynamic of HAV circulation was observed in the period post-outbreak; between 2017 and 2021 when 4 different lineages were transiently detected. Exhaustive phylogenetic analyses indicate that these lineages were introduced and possibly derived from isolates from other Latin American countries. CONCLUSION: The HAV circulation in recent years in Chile is rapidly changing and suggests that this phenomenon could be a consequence of massive population migrations in Latin America caused by political instability and natural disasters.

Trends and challenges in analytical chemistry for multi-analysis of illicit drugs employing wastewater-based epidemiology.

Wastewater-based epidemiology (WBE) for quantification of illicit drug biomarkers (IDBs) in wastewater samples is an effective tool that can provide information about drug consumption. The most commonly quantified IDBs belong to different chemical classes, including cocaine, amphetamine-type stimulants, opioids, and cannabinoids, so the different chemical properties of these molecules pose a challenge in the development of analytical methods for multi-analyte analysis. Recent workflows include the steps of sampling and storage, sample preparation using solid-phase extraction (SPE) or without extraction, and quantification of analytes employing gas or liquid chromatography coupled with mass spectrometry. The greatest difficulty is due to the fact that wastewater samples are complex chemical mixtures containing analytes with different chemical properties, often present at low concentrations. Therefore, in the development of analytical methods, there is the need to simplify and optimize the analytical workflows, reducing associated uncertainties, analysis times, and costs. The present work provides a critical bibliographic survey of studies published from the year 2020 until now, highlighting the challenges and trends of published analytical workflows for the multi-analysis of IDBs in wastewater samples, considering sampling and sample preparation, method validation, and analytical techniques.

Sewershed surveillance as a tool for smart management of a pandemic in threshold countries. Case study: Tracking SARS-CoV-2 during COVID-19 pandemic in a major urban metropolis in northwestern Argentina.

Wastewater-based epidemiology is an economical and effective tool for monitoring the COVID-19 pandemic. In this study we proposed sampling campaigns that addressed spatial-temporal trends within a metropolitan area. This is a local study of detection and quantification of SARS-CoV-2 in wastewater during the onset, rise, and decline of COVID-19 cases in Salta city (Argentina) over the course of a twenty-one-week period (13 Aug to 30 Dec) in 2020. Wastewater samples were gathered from 13 sewer manholes specific to each sewershed catchment, prior to convergence or mixing with other sewer lines, resulting in samples specific to individual catchments with defined areas. The 13 sewershed catchments selected comprise 118,832 connections to the network throughout the city, representing 84.7 % (534,747 individuals) of the total population. The number of COVID19-related exposure and symptoms cases in each area were registered using an application developed for smartphones by the provincial government. Geographical coordinates provided by the devices were recorded, and consequently, it was possible to geolocalise all app-cases and track them down to which of the 13 sampling catchments belonged. RNA fragments of SARS-CoV-2 were detected in every site since the beginning of the monitoring, anticipating viral circulation in the population. Over the course of the 21-week study, the concentrations of SARS-CoV-2 ranged between 1.77 × 104 and 4.35 × 107 genome copies/L. There was a correspondence with the highest viral load in wastewater and the peak number of cases reported by the app for each catchment. The associations were evaluated with correlation analysis. The viral loads of SARS-CoV-2 in wastewater were a feasible means to describe the trends of COVID-19 infections. Surveillance at sewershed scale, provided reliable and strategic information that could be used by local health stakeholders to manage the COVID-19 pandemic.

Quantification of SARS-CoV-2 in wastewater samples from hospitals treating COVID-19 patients during the first wave of the pandemic in Brazil.

The COVID-19 pandemic has caused a global health crisis, and wastewater-based epidemiology (WBE) has emerged as an important tool to assist public health decision-making. Recent studies have shown that the SARS-CoV-2 RNA concentration in wastewater samples is a reliable indicator of the severity of the pandemic for large populations. However, few studies have established a strong correlation between the number of infected people and the viral concentration in wastewater due to variations in viral shedding over time, viral decay, infiltration, and inflow. Herein we present the relationship between the number of COVID-19-positive patients and the viral concentration in wastewater samples from three different hospitals (A, B, and C) in the city of Belo Horizonte, Minas Gerais, Brazil. A positive and strong correlation between wastewater SARS-CoV-2 concentration and the number of confirmed cases was observed for Hospital B for both regions of the N gene (R = 0.89 and 0.77 for N1 and N2, respectively), while samples from Hospitals A and C showed low and moderate correlations, respectively. Even though the effects of viral decay and infiltration were minimized in our study, the variability of viral shedding throughout the infection period and feces dilution due to water usage for different activities in the hospitals could have affected the viral concentrations. These effects were prominent in Hospital A, which had the smallest sewershed population size, and where no correlation between the number of defecations from COVID-19 patients and viral concentration in wastewater was observed. Although we could not determine trends in the number of infected patients through SARS-CoV-2 concentrations in hospitals' wastewater samples, our results suggest that wastewater monitoring can be efficient for the detection of infected individuals at a local level, complementing clinical data.