Trends and Perspectives of Biosensors for Food and Environmental Virology

Food and environmental virology has become a very important and interesting area of research because of food safety and public health concerns. During the last few decades, increasing foodborne diseases and environmental generated illnesses are considered to be highly challenging issues. Biosensor technology holds great promise for the healthcare market, and the security sector. Similar to clinical diagnostic tools, biosensors are being developed for the rapid, reliable, yet inexpensive identification and enumeration of pathogenic viruses which are adulterating environment, food and feed commodities. In this modern era, bio-and nano-technologies play a pivotal role in virological diagnostics of food industry, environmental and veterinary samples. This review covers the recent advances and future prospects of nanotechnology-based bioanalytical microsystems for food and environmental virology.     

Norovirus GII.Pe Genotype: Tracking a Foodborne Outbreak on a Cruise Ship Through Molecular Epidemiology,Brazil, 2014

Norovirus (NoV) is recognized as the most common cause of foodborne outbreaks. In 2014, an outbreak of acute gastroenteritis occurred on a cruise ship in Brazil, and NoV became the suspected etiology. Here we present the molecular identification of the NoV strains and the use of sequence analysis to determine modes of virus transmission. Food (cream cheese, tuna salad, grilled fish, orange mousse, and vegetables soup) and clinical samples were analyzed by ELISA, conventional RT-PCR, qRT-PCR, and sequencing. Genogroup GII NoV was identified by ELISA and conventional RT-PCR in fecal samples from 5 of 12 patients tested (41.7%), and in the orange mousse food sample by conventional RT-PCR and qRT-PCR. Two fecal GII NoV samples and the orange mousse GII NoV sample were successfully genotyped as GII.Pe (ORF 1), revealed 98.0–98.8% identities among them, and shared phylogenetically distinct cluster. Establishing the source of a NoV outbreak can be a challenging task. In this report, the molecular analysis of the partial RdRp NoV gene provided a powerful tool for genotyping (GII.Pe) and tracking of outbreak-related samples. In addition, the same fast and simple extraction methods applied to clinical samples could be successfully used for complex food matrices, and have the potential to be introduced in routine laboratories for screening foods for presence of NoV.     

Survival and Inactivation by Advanced Oxidative Process of Foodborne Viruses in Model Low-Moisture Foods

Enteric viruses, such as human norovirus (NoV) and hepatitis A virus (HAV), are the major causes of foodborne illnesses worldwide. These viruses have low infectious dose, and may remain infectious for weeks in the environment and food. Limited information is available regarding viral survival and transmission in low-moisture foods (LMF). LMFs are generally considered as ready-to-eat products, which undergo no or minimal pathogen reduction steps. However, numerous foodborne viral outbreaks associated with LMFs have been reported in recent years. The objective of this study was to examine the survival of foodborne viruses in LMFs during 4-week storage at ambient temperature and to evaluate the efficacy of advanced oxidative process (AOP) treatment in the inactivation of these viruses. For this purpose, select LMFs such as pistachios, chocolate, and cereal were inoculated with HAV and the norovirus surrogates, murine norovirus (MNV) and feline calicivirus (FCV), then viral survival on these food matrices was measured over a four-week incubation at ambient temperature, by both plaque assay and droplet-digital RT-PCR (ddRT-PCR) using the modified ISO-15216 method as well as the magnetic bead assay for viral recovery. We observed an approximately 0.5 log reduction in viral genome copies, and 1 log reduction in viral infectivity for all three tested viruses following storage of select inoculated LMFs for 4 weeks. Therefore, the present study shows that the examined foodborne viruses can persist for a long time in LMFs. Next, we examined the inactivation efficacy of AOP treatment, which combines UV-C, ozone, and hydrogen peroxide vapor, and observed that while approximately 100% (4 log) inactivation can be achieved for FCV, and MNV in chocolate, the inactivation efficiency diminishes to approximately 90% (1 log) in pistachios and 70% (< 1 log) in cereal. AOP treatment could therefore be a good candidate for risk reduction of foodborne viruses from certain LMFs depending on the food matrix and surface of treatment.

     

Occurrence of Norovirus,Rotavirus, Hepatitis a Virus,and Enterovirus in Berries in Argentina

Food and Environmental Virology - Berries have been implicated as the probable vehicle of infection in multiple outbreaks of norovirus and hepatitis A virus (HAV). These foods often receive minimal...     

Virus Genome Quantification Does not Predict Norovirus Infectivity After Application of Food Inactivation Processing Technologies

When determining the effect of food processing on the infectivity of any contaminating virus, it is necessary to distinguish unambiguously between infectious and non-infectious viruses present. However, this can be difficult in the particular case of noroviruses (NoVs) because no reliable cell culture model is available. The aim of this study was to assess the use of molecular methods—RT real-time PCR (RT-qPCR) and enzymatic treatment (ET) coupled to RT-qPCR—to quantify the infectivity of NoV after application of various inactivating food-processing technologies. RT-qPCR and ET-RT-qPCR gave significantly different (P < 0.01) results concerning the reduction in viral genome counts by all inactivation procedures and conditions used, except for HHP treatment at 600 MPa for 5 min. These findings indicate that the ET prior to RT-qPCR has an effect on the estimation of the reduction of virus genome counts, and may eliminate genomes of affected virus particles. However, no correlation was found between the results obtained by ET-RT-qPCR and those obtained by cell culture. Therefore, the effect is presumably only partial, and not adequate to allow accurate estimation of virus inactivation. Consequently, our results indicate that the quantification of virus genomes by PCR, regardless of prior ET, is not adequate for establishing virus inactivation and/or infectivity. In addition, our results also illustrate that the general effect of virus inactivation is not directly correlated to effects on the integrity of virus genome and protein capsid. Presumably, inactivation by food processing is the consequence of effects on proteins involved in adhesion and invasion stages.     

A Review of Known and Hypothetical Transmission Routes for Noroviruses

Human noroviruses (NoVs) are considered a worldwide leading cause of acute non-bacterial gastroenteritis. Due to a combination of prolonged shedding of high virus levels in feces, virus particle shedding during asymptomatic infections, and a high environmental persistence, NoVs are easily transmitted pathogens. Norovirus (NoV) outbreaks have often been reported and tend to affect a lot of people. NoV is spread via feces and vomit, but this NoV spread can occur through several transmission routes. While person-to-person transmission is without a doubt the dominant transmission route, human infective NoV outbreaks are often initiated by contaminated food or water. Zoonotic transmission of NoV has been investigated, but has thus far not been demonstrated. The presented review aims to give an overview of these NoV transmission routes. Regarding NoV person-to-person transmission, the NoV GII.4 genotype is discussed in the current review as it has been very successful for several decades but reasons for its success have only recently been suggested. Both pre-harvest and post-harvest contamination of food products can lead to NoV food borne illness. Pre-harvest contamination of food products mainly occurs via contact with polluted irrigation water in case of fresh produce or with contaminated harvesting water in case of bivalve molluscan shellfish. On the other hand, an infected food handler is considered as a major cause of post-harvest contamination of food products. Both transmission routes are reviewed by a summary of described NoV food borne outbreaks between 2000 and 2010. A third NoV transmission route occurs via water and the spread of NoV via river water, ground water, and surface water is reviewed. Finally, although zoonotic transmission remains hypothetical, a summary on the bovine and porcine NoV presence observed in animals is given and the presence of human infective NoV in animals is discussed.     

Persistence of Human Noroviruses on Food Preparation Surfaces and Human Hands

The human noroviruses (NoV) are the major cause of acute non-bacterial gastroenteritis and are commonly transmitted by foodborne routes. Epidemiological evidence from propagated outbreaks, as well as environmental sampling, suggest that these viruses are environmentally stable. The purpose of this study was to examine the persistence of representative human NoV on the fingertips of volunteers and on commonly used food preparation surfaces. Human fingerpads and surfaces (stainless steel, Formica^®, and ceramic) were inoculated with 20% fecal suspensions of Norwalk virus (NV) or Snow Mountain virus (SMV). The virus inocula were recovered by elution at serial time points ranging from 0 to 120 min post-inoculation (for fingerpads) and after up to 42 days (for surfaces). The quantity of detectable viral RNA, expressed as genome equivalent particles (GEP) was evaluated using quantitative real-time RT-PCR (RT-qPCR). The amount of NV RNA on the surface decreased gradually over time, with an average reduction ranging from 1.5 to 2.9 log₁₀ GEP after 21–28 days storage under ambient conditions. SMV showed greater environmental persistence, with a 0.4–1.2 log₁₀ GEP reduction on all three surfaces after 42 days of ambient storage. On fingerpads, the amount of human NoV RNA declined slightly (<0.25 log₁₀) after 15 min and remained relatively unchanged thereafter (through 120 min). These results support the epidemiological evidence that food preparation surfaces and human hands can act as vehicles for human NoV transmission long after the initial contamination event has occurred.     

Control of Viral Contamination of Food and Environment

Viruses are often transmitted via food and the environment. Contamination may be controlled either by preventing its occurrence or by inactivating the contaminating virus. The majority of agents transmitted in this way are human enteric viruses, produced either in the intestines or the liver. They are shed in human feces (noroviruses also in vomitus) in a broad range of circumstances, and they are relatively stable outside the host. Non-enteric viruses are less often transmitted via foods and are generally less environmentally stable. Insofar as vaccines are available, they are able to prevent fecal shedding. Viruses shed in feces via the water-carriage toilet may be eliminated by proper treatment and disinfection of the wastewater. In the foods context, the most effective antiviral measures are cooking and hand washing. Detection methods are most useful after the fact, in investigating outbreaks and devising preventive measures.     

Norovirus Detection at Oualidia Lagoon,a Moroccan Shellfish Harvesting Area,by Reverse Transcription PCR Analysis

Norovirus (NoV) is the leading cause of acute viral gastroenteritis outbreaks in the world. These outbreaks are frequently associated with bivalve shellfish consumption, particularly because these products are often eaten raw or only slightly cooked. In Morocco, regulations concerning the acceptable levels of enteric bacteria indicator organisms in these products have been put in place. However, these regulations do not take into account the risk of viral contamination, and many gastroenteritis outbreaks have been linked to the ingestion of bivalve shellfish from areas that comply with the current food safety criteria. The aim of this study was to investigate NoV presence in shellfish samples (n = 104) collected at four sites owcff Oualidia lagoon (Moroccan Atlantic coast) from November 2015 to February 2017. Samples were analysed using real-time RT-PCR in accordance with the ISO 15216-2 method. NoVs of the genogroup II were detected in 7% of samples that were all collected during the winter months. Moreover, 71% of NoV-positive samples were harvested at sites upstream of the lagoon. These results highlight the need of regularly monitoring viral contamination in bivalve shellfish to limit the risk of viral gastroenteritis outbreaks.

     

Hepatitis A Virus: State of the Art

Hepatitis A is the most common among all hepatitis worldwide in spite of an efficient vaccine and improved hygiene. Shellfish-borne outbreaks are still of major concern causing hundreds of cases and huge economical losses in the present context of global food trade. Hepatitis A virus (HAV) is a unique picornavirus with many differences in its molecular biology including both its incapacity to induce the inhibition of the cellular protein synthesis and a highly biased and deoptimized codon usage with respect the cell. The final goal of this intriguing strategy seems to be the need for a fine-tuning control of the translation kinetics, particularly at the capsid coding region, and the underlying mechanism is the use of a right combination of common and rare codons to allow a regulated ribosome traffic rate thus ensuring the proper protein folding. Capsid folding is critical to warrant a high environmental stability for a virus transmitted through the fecal–oral route with long extracorporeal periods.     

Food security and global environmental change: emerging challenges

Most research linking global environmental change and food security focuses solely on agriculture: either the impact of climate change on agricultural production, or the impact of agriculture on the environment, e.g. on land use, greenhouse gas emissions, pollution and/or biodiversity. Important though food production is, many other factors also need to be considered to understand food security. A recent international conference on “Environmental Change and Food Security: Bridging Science, Policy and Development for Adaptation” included a range of papers that embraced the multiple dimensions of the food systems that underpin food security. The major conclusion from the conference was that technical fixes alone will not solve the food security challenge. Adapting to the additional threats to food security arising from major environmental changes requires an integrated food system approach, not just a focus on agricultural practices. Six key issues emerged for future research: (i) adapting food systems to global environmental change requires more than just technological solutions to increase agricultural yields; (ii) tradeoffs across multiple scales among food system outcomes are a pervasive feature of globalized food systems; (iii) within food systems, there are some key underexplored areas that are both sensitive to environmental change but also crucial to understanding its implications for food security and adaptation strategies; (iv) scenarios specifically designed to investigate the wider issues that underpin food security and the environmental consequences of different adaptation options are lacking; (v) price variability and volatility often threaten food security; and (vi) more attention needs to be paid to the governance of food systems.     

Enteric Viruses and Management of Shellfish Production in New Zealand

In New Zealand shellfish are a significant food resource and shellfish are harvested for both recreational and commercial use. Commercially harvested Greenshell mussels (Perna canaliculus) and Pacific oysters (Crassostrea gigas) from aquaculture farms dominate consumption in New Zealand. Other commercial species include cockles (Austrovenus stuchburyii) and surf clam species which are wild harvested. The consumption of shellfish has been associated with gastroenteritis outbreaks caused by noroviruses following faecal contamination of growing waters with human waste. In New Zealand, since 1994 over 50 norovirus outbreaks linked to consumption of either New Zealand commercially grown oysters or imported oysters have been reported. An IEC/ISO 17025 accredited method for detection of noroviruses in bivalve shellfish was established in 2007. This method has been used in outbreak investigations to analyse implicated shellfish, in virus prevalence surveys and monitoring programmes, and commercially for product clearances. Surveys have shown that enteric viruses occur frequently in non-commercial shellfish, especially near sewage outfalls and following sewage discharge events. Viral source tracking methods have assisted in identifying pollution sources. The commercial shellfish industry operates under the Bivalve Molluscan Shellfish Regulated Control Scheme (BMSRCS), administered by the New Zealand Food Safety Authority. Recently regulatory measures were introduced into the BMSRCS to manage viruses. These include the closure of harvest areas for at least 28 days after human sewage contamination events and norovirus outbreaks. These management strategies, coupled with new information on norovirus prevalence in shellfish, have helped to improve the quality and safety of New Zealand shellfish.     

The Fate of Murine Norovirus and Hepatitis A Virus During Preparation of Fresh Produce by Cutting and Grating

Human noroviruses and hepatitis A virus (HAV) are commonly associated with outbreaks occurring in restaurant establishments and catered events. Food handlers are major contributing factors to foodborne illnesses initiated in the kitchen setting. In this study, transfer of HAV and murine norovirus (MNV-1), a human norovirus surrogate, between produce (cucumbers, strawberries, tomatoes, cantaloupes, carrots, and honeydew melons) and common kitchen utensils (graters and knives) was investigated. The extent of virus transfer to produce during utensil application, in the presence and the absence of food residue, and the impact of knife surface properties (sharp, dull, serrated) was also investigated. Transfer of MNV-1 and HAV from produce items, initially contaminated with ~5.5 log PFU, to knives and graters during application ranged from 0.9 to 5.1 log PFU. MNV-1 transfer to knives was the greatest for cucumbers, strawberries, and tomatoes, and the least for honeydew melons, while transfer of HAV to knives was greater for tomatoes and honeydew melons than strawberries, cantaloupes, and cucumbers. After preparation of a contaminated produce item, knife cross-contamination easily occurred as viruses were detected on almost all of the seven produce items successively prepared. Produce residues on utensils often resulted in less virus transfer when compared to utensils without residue accumulation. Knife surface properties did not impact virus transfer. The ease of virus transfer between produce and utensils demonstrated by the current study highlights the importance of efforts aimed toward preventing cross-contamination in the kitchen environment.     

Critical Review of Norovirus Surrogates in Food Safety Research: Rationale for Considering Volunteer Studies

The inability to propagate human norovirus (NoV) or to clearly differentiate infectious from noninfectious virus particles has led to the use of surrogate viruses, like feline calicivirus (FCV) and murine norovirus-1 (MNV), which are propagatable in cell culture. The use of surrogates is predicated on the assumption that they generally mimic the viruses they represent; however, studies are proving this concept invalid. In direct comparisons between FCV and MNV, their susceptibility to temperatures, environmental and food processing conditions, and disinfectants are dramatically different. Differences have also been noted between the inactivation of NoV and its surrogates, thus questioning the validity of surrogates. Considerable research funding is provided globally each year to conduct surrogate studies on NoVs; however, there is little demonstrated benefit derived from these studies in regard to the development of virus inactivation techniques or food processing strategies. Human challenge studies are needed to determine which processing techniques are effective in reducing NoVs in foods. A major obstacle to clinical trials on NoVs is the perception that such trials are too costly and risky, but in reality, there is far more cost and risk in allowing millions of unsuspecting consumers to contract NoV illness each year, when practical interventions are only a few volunteer studies away. A number of clinical trials have been conducted, providing important insights into NoV inactivation. A shift in research priorities from surrogate research to volunteer studies is essential if we are to identify realistic, practical, and scientifically valid processing approaches to improve food safety.     

Norovirus Foodborne Outbreak Associated With the Consumption of Ice Pop,Southern Brazil, 2020

Food and Environmental Virology - Norovirus is a major cause of foodborne-associated acute gastroenteritis (AGE) outbreaks worldwide. Usually, food products are contaminated either during...     

New Proof-of-Concept in Viral Inactivation: Virucidal Efficacy of 405 nm Light Against Feline Calicivirus as a Model for Norovirus Decontamination

The requirement for novel decontamination technologies for use in hospitals is ever present. One such system uses 405 nm visible light to inactivate microorganisms via ROS-generated oxidative damage. Although effective for bacterial and fungal inactivation, little is known about the virucidal effects of 405 nm light. Norovirus (NoV) gastroenteritis outbreaks often occur in the clinical setting, and this study was designed to investigate potential inactivation effects of 405 nm light on the NoV surrogate, feline calicivirus (FCV). FCV was exposed to 405 nm light whilst suspended in minimal and organically-rich media to establish the virucidal efficacy and the effect biologically-relevant material may play in viral susceptibility. Antiviral activity was successfully demonstrated with a 4 Log₁₀ (99.99%) reduction in infectivity when suspended in minimal media evident after a dose of 2.8 kJ cm^?2. FCV exposed in artificial faeces, artificial saliva, blood plasma and other organically rich media exhibited an equivalent level of inactivation using between 50–85% less dose of the light, indicating enhanced inactivation when the virus is present in organically-rich biologically-relevant media. Further research in this area could aid in the development of 405 nm light technology for effective NoV decontamination within the hospital environment.     

渤海湾天津沿岸海水中甲肝病毒的检测和定量

甲肝病毒（hepatitis A virus, HAV）是能引起传染性甲型肝炎的单链RNA病毒.用常规RT-PCR（反转录PCR）和SYBR Green实时定量RT-PCR方法,根据甲肝病毒保守的VP1-VP2基因序列设计引物,对渤海湾天津沿岸海水中的甲肝病毒进行了检测和定量分析. 9个样品取自渤海湾天津塘沽以南沿岸海水,取样时间分别是2007年夏、秋、冬季和2008年春季. 海水样品先用小型超滤装置（Millipore Pellicon Mini TFF）或超滤离心管（Millipore Centricon Plus-70）浓缩,然后进行RT-PCR检测.结果表明,从9个海水样品中都能扩增出192 bp的HAV cDNA,这些cDNA的核苷酸序列与GenBank中的同源序列相似性为95%～100%. 用SYBR Green 实时定量RT-PCR检测了春季和冬季的6个海水样品,结果表明,海水中甲肝病毒的浓度范围为5.35×10⁶～4.51×10⁷ virus particles/L.     

国外食物系统研究综述及借鉴

食物关系到国计民生,食物的高效供给取决于食物系统的高效运转。目前国内食物系统视角的研究不足,系统梳理国外研究进展对我国食物系统研究具有积极的借鉴意义。为了更好地把握食物系统的研究脉络,论文通过对国外食物系统研究文献进行研读,在总结分析食物系统研究相关理论和方法的基础上,着重对食物系统的全球化与本地化、食物系统规划、食物系统的资源环境效应、食物安全与政策研究等问题进行了系统梳理。在借鉴国外食物系统研究的基础上,结合已有的研究基础和社会需求,提出未来我国食物系统领域的研究展望,包括开展不同尺度的食物系统实证研究、开展跨学科多领域的共同研究、食物系统定量指标体系设计问题研究、食物系统的利益相关者研究以及加强技术创新对食物系统的影响研究等。