A review on mpox (monkeypox) virus shedding in wastewater and its persistence evaluation in environmental samples

Since May 2022, an outbreak of mpox (formerly known as monkeypox) has been spreading in non-endemic countries and this disease is caused by a mpox virus (MPXV) belonging to Orthopoxvirus genus of the Poxviridae family. Public health surveillance is essential to assess the disease prevalence and spread in the populations. In this purpose, wastewater surveillance has proved to be an effective and affordable tool to detect and monitor disease threats and outbreaks, such as SARS-CoV-2, poliovirus, and other viruses. Prior to the current mpox outbreak, previous studies suggested that MPXV shedding is possible after viral genome and infectious particles were detected in the feces of infected animals. On the other hand, using metagenomic and sequencing approaches to assess the diversity of human viral pathogens in wastewater, contigs assigned to orthopoxviruses and poxviruses were among the most abundant contigs detected in collected samples. Recently, MPXV DNA has been detected in wastewater samples from wastewater treatment plants in cities and airports in the United States, the Netherlands, Italy, France, and Spain. It has long been reported that poxviruses are stable in the environment for long periods and might represent as a source of transmission. However, the stability of MPXV in the environment is unknown and the presence of infectious MPXV in wastewater has not yet been determined. The present work is devoted to reviewing past and recent advances in MPXV detection in wastewater. It also addressed the survival data of orthopoxviruses, which may be of interest to evaluate the persistence of MPXV species in environmental matrices.

Assessment of Fecal Viral Shedding Among Children Who Have COVID-19 by Polymerase Chain Reaction

Objective: This study aimed to assess fecal viral shedding in children who have been confirmed COVID-19 by real time polymerase chain reaction (RT-PCR). Material(s) and Method(s): We enrolled fifty inpatient children who have been confirmed COVID-19 during first wave of outbreak in our region. All of the patients have been twice confirmed by RT-PCR within nasopharyngeal swabs. Each case was evaluated with clinical data, laboratory tests and rectal swabs. The rectal swabs were obtained five days after nasopharyngeal positivity. The clinical data was recorded within two basic categories, including common symptoms or digestive symptoms. Detection of SARS-CoV-2 in rectal swabs was performed by RT-PCR method. Result(s): Fifteen patients (30%) had digestive symptoms. On the 5th day, 50 rectal swabs were studied with PCR-RT. Only one of them was positive (2%). The only patient who was positive for SARS-CoV-2 on rectal swab was a symptomatic threeand-a-half-year-old girl. The patient, who became asymptomatic in the follow-up, was retested with a nasopharyngeal swab one week later, the result was negative and she was discharged on the 10th day. The second rectal smear of the patient was negative. Conclusion(s): We found very low rate (2%) fecal viral shedding with rectal swab PCR among children who have been confirmed COVID-19 by nasopharyngeal swab PCR. We thought that this result was due to the mild clinical course of the patients who has been diagnosed with COVID-19 we followed up. However, fecal oral transmission might still matter in children. Copyright © 2022 Ankara Pediatric Hematology Oncology Training and Research Hospital. All rights reserved.

Population level SARS-CoV-2 fecal shedding rates determined via wastewater-based epidemiology.

Wastewater-based epidemiology (WBE) has been utilized as an early warning tool to anticipate disease outbreaks, especially during the COVID-19 pandemic. However, COVID-19 disease models built from wastewater-collected data have been limited by the complexities involved in estimating SARS-CoV-2 fecal shedding rates. In this study, wastewater from six municipalities in Arizona and Florida with distinct demographics were monitored for SARS-CoV-2 RNA between September 2020 and December 2021. Virus concentrations with corresponding clinical case counts were utilized to estimate community-wide fecal shedding rates that encompassed all infected individuals. Analyses suggest that average SARS-CoV-2 RNA fecal shedding rates typically occurred within a consistent range (7.53-9.29 log10 gc/g-feces); and yet, were unique to each community and influenced by population demographics. Age, ethnicity, and socio-economic factors may have influenced shedding rates. Interestingly, populations with median age between 30 and 39 had the greatest fecal shedding rates. Additionally, rates remained relatively constant throughout the pandemic provided conditions related to vaccination and variants were unchanged. Rates significantly increased in some communities when the Delta variant became predominant. Findings in this study suggest that community-specific shedding rates may be appropriate in model development relating wastewater virus concentrations to clinical case counts.

Quantifying the Relationship between SARS-CoV-2 Wastewater Concentrations and Building-Level COVID-19 Prevalence at an Isolation Residence: A Passive Sampling Approach.

SARS-CoV-2 RNA loads can be detected in the excreta of individuals with COVID-19 and have demonstrated positive correlations with clinical infection trends. Consequently, wastewater-based epidemiology (WBE) approaches have been implemented globally as a public health surveillance tool to monitor community-level prevalence of infections. The majority of wastewater specimens are gathered as either composite samples via automatic samplers (autosamplers) or grab samples. However, autosamplers are expensive and can be challenging to maintain in cold weather, while grab samples are particularly susceptible to temporal variation when sampling sewage directly from complex matrices outside residential buildings. Passive sampling can provide an affordable, practical, and scalable sampling system while maintaining a reproducible SARS-CoV-2 signal. In this regard, we deployed tampons as passive samplers outside of a COVID-19 isolation unit (a segregated residence hall) at a university campus from 1 February 2021-21 May 2021. Samples (n = 64) were collected 3-5 times weekly and remained within the sewer for a median duration of 24 h. SARS-CoV-2 RNA was quantified using reverse-transcription quantitative polymerase chain reaction (RT-qPCR) targeting the N1 and N2 gene fragments. We quantified the mean viral load captured per individual and the association between the daily viral load and total persons, adjusting for covariates using multivariable models to provide a baseline estimate of viral shedding. Samples were processed through two distinct laboratory pipelines on campus, yielding highly correlated N2 concentrations. Data obtained here highlight the success of passive sampling utilizing tampons to capture SARS-CoV-2 in wastewater coming from a COVID-19 isolation residence, indicating that this method can help inform building-level public health responses.

Enumerating asymptomatic COVID-19 cases and estimating SARS-CoV-2 fecal shedding rates via wastewater-based epidemiology.

Wastewater-based epidemiology (WBE) was utilized to monitor SARS-CoV-2 RNA in sewage collected from manholes specific to individual student dormitories (dorms) at the University of Arizona in the fall semester of 2020, which led to successful identification and reduction of SARS-CoV-2 transmission events. Positive wastewater samples triggered clinical testing of residents within that dorm; thus, SARS-CoV-2 infected individuals were identified regardless of symptom expression. This current study examined clinical testing data to determine the abundance of asymptomatic versus symptomatic cases in these defined communities. Nasal and nasopharyngeal swab samples processed via antigen and PCR tests indicated that 79.2% of SARS-CoV-2 infections were asymptomatic, and only 20.8% of positive cases reported COVID-19 symptoms at the time of testing. Clinical data was paired with corresponding wastewater virus concentrations, which enabled calculation of viral shedding rates in feces per infected person. Mean shedding rates averaged from positive wastewater samples across all dorms were 7.30 ± 0.67 log10 genome copies per gram of feces (gc/g-feces) based on the N1 gene. Quantification of SARS-CoV-2 fecal shedding rates from infected individuals has been the critical missing component necessary for WBE models to measure and predict SARS-CoV-2 infection prevalence in communities. The findings from this study can be utilized to create models that can be used to inform public health prevention and response actions.

Early warning of COVID-19 via wastewater-based epidemiology: potential and bottlenecks.

An effective early warning tool is of great administrative and social significance to the containment and control of an epidemic. Facing the unprecedented global public health crisis caused by COVID-19, wastewater-based epidemiology (WBE) has been given high expectations as a promising surveillance complement to clinical testing which had been plagued by limited capacity and turnaround time. In particular, recent studies have highlighted the role WBE may play in being a part of the early warning system. In this study, we briefly discussed the basics of the concept, the benefits and critical points of such an application, the challenges faced by the scientific community, the progress made so far, and what awaits to be addressed by future studies to make the concept work. We identified that the shedding dynamics of infected individuals, especially in the form of a mathematical shedding model, and the back-calculation of the number of active shedders from observed viral load are the major bottlenecks of WBE application in the COVID-19 pandemic that deserve more attention, and the sampling strategy (location, timing, and interval) needs to be optimized to fit the purpose and scope of the WBE project.

Candidate Gene Markers Associated with Fecal Shedding of the Feline Enteric Coronavirus (FECV).

The Feline coronavirus (FCoV) can cause a fatal disease, the Feline Infectious Peritonitis. Persistent shedders represent the most important source of infection. The role of the host in FCoV fecal shedding is unknown. The objective of this study was to develop gene markers and to test their associations with FCoV shedding patterns. Fecal samples were taken from 57 cats of 12 breeds on the day 0 and after 2, 4 and 12 months. Variation from persistent and/or high-intensity shedding to no shedding was observed. Thirteen immunity-related genes were selected as functional and positional/functional candidates. Positional candidates were selected in a candidate region detected by a GWAS analysis. Tens to hundreds of single nucleotide polymorphisms (SNPs) per gene were identified using next generation sequencing. Associations with different phenotypes were assessed by chi-square and Fisher's exact tests. SNPs of one functional and one positional candidate (NCR1 and SLX4IP, respectively) and haplotypes of four genes (SNX5, NCR2, SLX4IP, NCR1) were associated with FCoV shedding at pcorected < 0.01. Highly significant associations were observed for extreme phenotypes (persistent/high-intensity shedders and non-shedders) suggesting that there are two major phenotypes associated with different genotypes, highly susceptible cats permanently shedding high amounts of viral particles and resistant non-shedders.

Prolonged viral shedding in feces of pediatric patients with coronavirus disease 2019.

OBJECTIVE: To determine the dynamic changes of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in respiratory and fecal specimens in children with coronavirus disease 2019 (COVID-19). METHODS: From January 17, 2020 to February 23, 2020, three paediatric cases of COVID-19 were reported in Qingdao, Shandong Province, China. Epidemiological, clinical, laboratory, and radiological characteristics and treatment data were collected. Patients were followed up to March 10, 2020, and dynamic profiles of nucleic acid testing results in throat swabs and fecal specimens were closely monitored. RESULTS: Clearance of SARS-CoV-2 in respiratory tract occurred within two weeks after abatement of fever, whereas viral RNA remained detectable in stools of pediatric patients for longer than 4 weeks. Two children had fecal SARS-CoV-2 undetectable 20 days after throat swabs showing negative, while that of another child lagged behind for 8 days. CONCLUSIONS: SARS-CoV-2 may exist in children's gastrointestinal tract for a longer time than respiratory system. Persistent shedding of SARS-CoV-2 in stools of infected children raises the possibility that the virus might be transmitted through contaminated fomites. Massive efforts should be made at all levels to prevent spreading of the infection among children after reopening of kindergartens and schools.