Optimisation of a PMAxx™-RT-qPCR Assay and the Preceding Extraction Method to Selectively Detect Infectious Murine Norovirus Particles in Mussels

Human noroviruses are a major cause for gastroenteritis outbreaks. Filter-feeding bivalve molluscs, which accumulate noroviruses in their digestive tissues, are a typical vector for human infection. RT-qPCR, the established method for human norovirus detection in food, does not allow discrimination between infectious and non-infectious viruses and can overestimate potentially infectious viral loads. To develop a more accurate method of infectious norovirus load estimation, we combined intercalating agent propidium monoazide (PMAxx™)-pre-treatment with RT-qPCR assay using in vitro-cultivable murine norovirus. Three primer sets targeting different genome regions and diverse amplicon sizes were used to compare one-step amplification of a short genome fragment to three two-step long-range RT-qPCRs (7 kbp, 3.6 kbp and 2.3 kbp amplicons). Following initial assays performed on untreated infectious, heat-, or ultraviolet-inactivated murine noroviruses in PBS suspension, PMAxx™ RT-qPCRs were implemented to detect murine noroviruses subsequent to their extraction from mussel digestive tissues; virus extraction via anionic polymer-coated magnetic beads was compared with the proteinase K-dependent ISO norm. The long-range RT-qPCR process detecting fragments of more than 2.3 kbp allowed accurate estimation of the infectivity of UV-damaged murine noroviruses. While proteinase K extraction limited later estimation of PMAxx™ pre-treatment effects and was found to be unsuited to the assay, magnetic bead-captured murine noroviruses retained their infectivity. Genome copies of heat-inactivated murine noroviruses differed by 2.3 log₁₀ between RT-qPCR and PMAxx™-RT-qPCR analysis in bivalve molluscs, the PMAxx™ pre-treatment allowing a closer approximation of infectious titres. The combination of bead-based virus extraction and PMAxx™ RT-qPCR thus provides a more accurate model for the estimation of noroviral bivalve mollusc contamination than the conjunction of proteinase K extraction and RT-qPCR and has the potential (once validated utilising infectious human norovirus) to provide an added measure of security to food safety authorities in the hazard assessment of potential bivalve mollusc contamination.

     

Assessment of Viral Contamination of Five Brazilian Artisanal Cheese Produced from Raw Milk: a Randomized Survey

Food and Environmental Virology - Enteric viruses have been described as important contaminants in fresh and ready-to-eat foods such as sandwiches, deli meat and dairy products. This is a...     

The impact of COVID-19 pandemic in medical waste amounts: a case study from a high-populated city of Turkey

Journal of Material Cycles and Waste Management - In this study, the amount of medical waste generated in a high-populated city in Turkey between January 2018 and August 2021 was evaluated, and the...     

The combination of chemical,structural, and functional indicators to evaluate the anthropogenic impacts on agricultural stream ecosystems

Environmental Science and Pollution Research - Freshwater contamination by pesticides in agricultural landscapes is of increasing concern worldwide, with strong pesticide impacts on biodiversity,...     

Environmental and social impacts of the increasing number of geothermal power plants (Büyük Menderes Graben—Turkey)

Environmental Science and Pollution Research - The use of renewable energy is critical to the long-term development of global energy. Geothermal power plants (GPPs) differ in the technology they...     

An empirical note on tourism and sustainable development nexus

Environmental Science and Pollution Research - The goal of this research is to investigate the impact of tourism on sustainable development in the 10 most visited countries. For this purpose,...     

Are sea snakes pertinent bio-indicators for coral reefs? a comparison between species and sites

Classical sampling methods often miss important components of coral reef biodiversity, notably organisms that remain sheltered within the coral matrix. Recent studies using sea kraits (sea snakes) as bio-indicators suggest that the guild of predators represented by anguilliform fish (Congridae, Muraenidae, Ophichthidae, henceforth “eels” for simplicity) were far more abundant and diverse than previously suspected. In the current study, eel diversity (similarity and species richness indices) estimated via sea snake sampling (SSS) was compared among six areas of one of the main oceanic biodiversity hotspot of the Pacific Ocean (southwest lagoon of New Caledonia). Based on the eel diversity in the snakes’ diet, the results obtained in six areas, in two snake species, and using different estimates (ANOSIM, Shannon index…) were consistent, suggesting that SSS provided robust information. Analyses also suggested subtle, albeit significant, differences in the eel assemblages among islets. Such spatial differences are discussed in light of local management practices. As SSS is easy to use, cost-effective, and provides the best picture of eel assemblages to date, it can be employed to monitor the eel assemblages in addition to the snakes themselves in many areas of the Indo-Pacific Ocean, thereby providing an index of the top predator biodiversity of many coral reefs.     

Composition of plastics from waste electrical and electronic equipment (WEEE) by direct sampling

This paper describes a direct analysis study carried out in a recycling unit for waste electrical and electronic equipment (WEEE) in Portugal to characterize the plastic constituents of WEEE. Approximately 3400 items, including cooling appliances, small WEEE, printers, copying equipment, central processing units, cathode ray tube (CRT) monitors and CRT televisions were characterized, with the analysis finding around 6000 kg of plastics with several polymer types. The most common polymers are polystyrene, acrylonitrile-butadiene-styrene, polycarbonate blends, high-impact polystyrene and polypropylene. Additives to darken color are common contaminants in these plastics when used in CRT televisions and small WEEE. These additives can make plastic identification difficult, along with missing polymer identification and flame retardant identification marks. These drawbacks contribute to the inefficiency of manual dismantling of WEEE, which is the typical recycling process in Portugal. The information found here can be used to set a baseline for the plastics recycling industry and provide information for ecodesign in electrical and electronic equipment production.     

Aerobic Biodegradation of Polymers in Solid-State Conditions: A Review of Environmental and Physicochemical Parameter Settings in Laboratory Simulations

During the last few years, biodegradable polymers have been developed to replace petrochemical polymers. Until now, research devoted to these polymers essentially focused on their biodegradability. There is now a need to bear out their nontoxicity. To verify this, the biodegradation must be carried out in accelerated laboratory tests which allow the metabolites and residues to be recovered. To reproduce the natural conditions (compost, field) as closely as possible, degradation experiments must be run on solid-state substrates. We review studies of aerobic degradation in solid-state substrates. This article focuses in particular on the environmental, physical, and chemical parameters (such as substrate nature, moisture, temperature, C/N ratio, and pH) that influence biodegradation kinetics. This study also aims at finding the solid substrate most adapted to residues and metabolite recovery. The most significant parameters would appear to be the substrate type, moisture content, and temperature. Inert substrates such as vermiculite are well suited to residue extraction. This review also opens the field to new research aimed at optimizing conditions for aerobic solid-state biodegradation and at recovering the metabolites and residues of this degradation process.     

Megaprojects and social and environmental changes: The case of the Thai "water grid"

Large-scale development of irrigation has long been an attractive option of postwar development, and the Mekong region has been no exception. Thailand has developed approximately four million hectares of irrigated land, and its northeastern region (Isaan)--both the driest and poorest part of the country--has been the target of many water projects. However, "full development" of its potential has been constrained by the lack of storage sites and the difficulty of diverting water from the Mekong River. Several ambitious projects have been discussed during the last 50 y, all of which have been aimed at "greening Isaan." In 2003, the Thai administration launched the idea of a national "water grid" that would triple the area of irrigated land in the country. This paper analyzes the emergence of this megaproject, its governance, and its economic and environmental soundness.