Investigating Non-Native Ribbon Worm as a Potential Source of Tetrodotoxin in British Bivalve Shellfish.

Tetrodotoxin (TTX) is a potent marine neurotoxin found in several phylogenetically diverse organisms, some of which are sought as seafood. Since 2015, TTX has been reported in bivalve shellfish from several estuarine locations along the Mediterranean and European Atlantic coasts, posing an emerging food safety concern. Although reports on spatial and temporal distribution have increased in recent years, processes leading to TTX accumulation in European bivalves are yet to be described.

Realising a global One Health disease surveillance approach: insights from wastewater and beyond.

One Health is a recognition of the shared environment inhabited by humans, animals and plants, and the impact of their interactions on the health of all organisms. The COVID-19 pandemic highlighted the need for a framework of pathogen surveillance in a tractable One Health paradigm to allow timely detection and response to threats to human and animal health. We present case studies centered around the recent global approach to tackle antimicrobial resistance and the current interest in wastewater testing, with the concept of "one sample many analyses" to be further explored as the most appropriate means of initiating this endeavor.

An international inter-laboratory study to compare digital PCR with ISO standardized qPCR assays for the detection of norovirus GI and GII in oyster tissue.

An optimized digital RT-PCR (RT-dPCR) assay for the detection of human norovirus GI and GII RNA was compared with ISO 15216-conform quantitative real-time RT-PCR (RT-qPCR) assays in an interlaboratory study (ILS) among eight laboratories. A duplex GI/GII RT-dPCR assay, based on the ISO 15216-oligonucleotides, was used on a Bio-Rad QX200 platform by six laboratories. Adapted assays for Qiagen Qiacuity or ThermoFisher QuantStudio 3D were used by one laboratory each.

Development and validation of a duplex RT-qPCR assay for norovirus quantification in wastewater samples.

Norovirus (NoV) is a highly contagious enteric virus that causes widespread outbreaks and a substantial number of deaths across communities. As clinical surveillance is often insufficient, wastewater-based epidemiology (WBE) may provide novel pathways of tracking outbreaks. To utilise WBE, it is important to use accurate and sensitive methods for viral quantification.

The Inhibition and Variability of Two Different RT-qPCR Assays Used for Quantifying SARS-CoV-2 RNA in Wastewater.

Faecal shedding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its subsequent detection in wastewater turned the spotlight onto wastewater-based epidemiology (WBE) for monitoring the coronavirus-disease 2019 (COVID-19) pandemic. WBE for SARS-CoV-2 has been deployed in 70 countries, providing insights into disease prevalence, forecasting and the spatiotemporal tracking and emergence of SARS-CoV-2 variants. Wastewater, however, is a complex sample matrix containing numerous reverse transcription quantitative PCR (RT-qPCR) inhibitors whose concentration and diversity are influenced by factors including population size, surrounding industry and agriculture and climate.

An analysis of 45 large-scale wastewater sites in England to estimate SARS-CoV-2 community prevalence.

Accurate surveillance of the COVID-19 pandemic can be weakened by under-reporting of cases, particularly due to asymptomatic or pre-symptomatic infections, resulting in bias. Quantification of SARS-CoV-2 RNA in wastewater can be used to infer infection prevalence, but uncertainty in sensitivity and considerable variability has meant that accurate measurement remains elusive. Here, we use data from 45 sewage sites in England, covering 31% of the population, and estimate SARS-CoV-2 prevalence to within 1.

Correction: Farkas et al. Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR. 2021, , 17.

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Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR.

Wastewater-based epidemiology has become an important tool for the surveillance of SARS-CoV-2 outbreaks. However, the detection of viruses in sewage is challenging and to date there is no standard method available which has been validated for the sensitive detection of SARS-CoV-2. In this paper, we describe a simple concentration method based on polyethylene glycol (PEG) precipitation, followed by RNA extraction and a one-step quantitative reverse transcription PCR (qRT-PCR) for viral detection in wastewater.

A qPCR-MPN method for rapid quantification of Escherichia coli in bivalve molluscan shellfish.

Faecal contamination of bivalve molluscan shellfish (BMS) can lead to infections from enteric pathogens if consumed. Across Europe, the faecal indicator bacteria Escherichia coli, is used to determine contamination of BMS harvesting areas. The reference most probable number (MPN) method for E.

Strategies to reduce norovirus (NoV) contamination from oysters under depuration conditions.

Depuration of oysters can effectively reduce levels of E. coli, however, may not be effective in safeguarding against viral contamination (EFSA, 2012). These trials assess the removal of Norovirus Genogroups I and II (NoV GI and GII) and F + RNA bacteriophage genogroup II (FRNAP-II) from oysters under depuration using molecular and viability assay methods.

Whole Genome Sequencing of Hepatitis A Virus Using a PCR-Free Single-Molecule Nanopore Sequencing Approach.

Hepatitis A virus (HAV) is one of the most common causes of acute viral hepatitis in humans. Although HAV has a relatively small genome, there are several factors limiting whole genome sequencing such as PCR amplification artefacts and ambiguities in assembly. The recently developed Oxford Nanopore technologies (ONT) allows single-molecule sequencing of long-size fragments of DNA or RNA using PCR-free strategies.

Long term reductions of faecal indicator organisms in Chichester Harbour (England) following sewerage infrastructure improvements in the catchment.

Quantitative information relating sewerage infrastructure schemes to microbial water quality improvements in recreational and shellfish harvesting areas is lacking. In this study, we assessed the effect of two sewerage schemes on concentrations of faecal indicator organisms (FIO) in Chichester Harbour, an important oyster fishery and water recreation area in the UK. The sewerage schemes comprised the installation of activated sludge and UV disinfection plants and increase in the storage capacity of storm tanks at sewage treatment works that discharge to tidal waters.

Viral indicators for tracking domestic wastewater contamination in the aquatic environment.

Waterborne enteric viruses are an emerging cause of disease outbreaks and represent a major threat to global public health. Enteric viruses may originate from human wastewater and can undergo rapid transport through aquatic environments with minimal decay. Surveillance and source apportionment of enteric viruses in environmental waters is therefore essential for accurate risk management.

Adolescents' application of the virtues across five cultural contexts.

Little is known about adolescent applications of the virtues such as honesty, responsibility and courage across different cultural contexts. Using the Adolescent Intermediate Concepts Measure we analyze samples of adolescents (ages 12 to 20; N = 9,112) from 5 contexts: North Macedonia, Mexico, Taiwan, the United Kingdom, and the United States. Across samples, adolescents provide evidence of developmental growth in the ability to apply virtue concepts as assessed by responses to dilemma-based situations.

Assessment of the Applicability of Capsid-Integrity Assays for Detecting Infectious Norovirus Inactivated by Heat or UV Irradiation.

Human noroviruses are the leading cause of viral gastroenteritis. In the absence of a practical culture technique for routine analysis of infectious noroviruses, several methods have been developed to discriminate between infectious and non-infectious viruses by removing non-viable viruses prior to analysis by RT-qPCR. In this study, two such methods (RNase and porcine gastric mucin) which were designed to remove viruses with compromised capsids (and therefore assumed to be non-viable), were assessed for their ability to quantify viable F-specific RNA bacteriophage (FRNAP) and human norovirus following inactivation by UV-C or heat.

Critical Evaluation of CrAssphage as a Molecular Marker for Human-Derived Wastewater Contamination in the Aquatic Environment.

The discharge of human-derived wastewater represents a major threat to water quality with the potential for waterborne disease outbreaks mainly associated with enteric viruses. To prevent illnesses, indicators associated with fecal contamination are monitored in polluted areas, however, their prevalence often does not correlate well with viral pathogens. In this study, we used crAssphage, a recently discovered human-specific gut-associated bacteriophage, for the surveillance of wastewater-derived viral contamination.

Escherichia coli testing and enumeration in live bivalve shellfish - Present methods and future directions.

A wide variety of pathogenic agents such as bacteria, viruses and parasites can be greatly concentrated in filter feeding bivalve molluscan shellfish (BMS), that are grown in faecally contaminated waters. Human health risks associated with the consumption of BMS are also compounded by the traditional pattern of consuming them raw or lightly cooked. Because of these well-established food safety risks, food legislation such as that in Europe stipulates that BMS production areas are monitored for faecal contamination and classified accordingly.

A highly specific Escherichia coli qPCR and its comparison with existing methods for environmental waters.

The presence of Escherichia coli in environmental waters is considered as evidence of faecal contamination and is therefore commonly used as an indicator in both water quality and food safety analysis. The long period of time between sample collection and obtaining results from existing culture based methods means that contamination events may already impact public health by the time they are detected. The adoption of molecular based methods for E.

Effectiveness of a multimedia outreach kit for families of wounded veterans.

Background: Young children in military families with a member who has a life changing injury can experience emotional difficulties and behavior changes.

Objective: This study evaluated a Sesame Workshop multimedia kit called: Talk, Listen, Connect: Changes (TLC-II(C); 2008). The kit, which included video and print materials, aimed to help caregivers (i.