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Microplastics in aquatic ecosystems harbor numerous microorganisms, including pathogenic species. The ingestion of these microplastics by commercial fish poses a threat to the ecosystem and human livelihood. Coastal lagoons are highly vulnerable to microplastic and microbiological pollution, yet limited understanding of the risks complicates management. Here, we present the main bacterial groups, including potentially pathogenic species, identified on microplastics in waters, sediments, and commercial fish from Ciénaga Grande de Santa Marta (CGSM), the largest coastal lagoon in Colombia. DNA metabarcoding allowed identifying 1760 bacterial genera on microplastics, with Aeromonas and Acinetobacter as the most frequent and present in all three matrices. The greatest bacterial richness and diversity were recorded on microplastics from sediments, followed by waters and fish. Biochemical analyses yielded 19 species of potentially pathogenic culturable bacteria on microplastics. Aeromonas caviae was the most frequent and, along with Pantoea sp., was found on microplastics in all three matrices. Enterobacter roggenkampii and Pseudomonas fluorescens were also found on microplastics from waters and fish. We propose management strategies for an Early Warning System against microbiological and microplastic pollution risks in coastal lagoons, illustrated by CGSM. This includes forming inter-institutional alliances for research and monitoring, accompanied by strengthening governance and health infrastructures.

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In the context of climate change and depletion of natural resources, meeting the growing demand for animal feed and human food through sufficient, nutritious, safe, and affordable sources of protein is becoming a priority. The use of Hermetia illucens, the black soldier fly (BSF), has emerged as a strategy to enhance the circularity of the agri-food chain, but its microbiological safety remains a concern. The aim of the present study was to systematically review available data on the microbiological quality of BSF and to investigate the impact of using four different rearing substrates including classic options allowed by the EU regulation (cereals, fruits, vegetables) and options not allowed by EU regulations regarding vegetable agri-food (co-products, food at shelf life, and meat). A total of 13 studies were collected and synthesized, including 910 sample results, while 102 new sample results were collected from the present experiments in three farms. Both datasets combined revealed a high level of contamination of larvae, potentially transmitted through the substrate. The main pathogenic bacteria identified were Bacillus cereus, Clostridium perfringens, Cronobacter spp., Escherichia coli, Salmonella spp., and Staphylococcus aureus coagulase-positive, while Campylobacter spp. and Listeria monocytogenes were not detected. Any of these four substrates were excluded for their use in insect rearing; however, safety concerns were confirmed and must be managed by the operators of the sector using microbial inactivation treatment after the harvest of the larvae in order to propose safe products for the market. The results obtained will guide the definition of the control criteria and optimize the following manufacturing steps.

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Organic farming is a sustainable agricultural practice emphasizing natural inputs and ecological balance, and has garnered significant attention for its potential health and environmental benefits. However, a comprehensive evaluation of the emergent contaminants, particularly resistance and virulence genes within organic farming system, remains elusive. Here, a total of 36 soil samples from paired conventional and organic vegetable farms were collected from Jiangsu province, China. A systematic metagenomic approach was employed to investigate the prevalence, dispersal capability, pathogenic potential, and drivers of resistance and virulence genes across both organic and conventional systems. Our findings revealed a higher abundance of antibiotic resistance genes (ARGs), biocide resistance genes (BRGs), and virulence factor genes (VFGs) in organic farming system, with ARGs exhibiting a particularly notable increase of 10.76% compared to the conventional one. Pathogens such as Pseudomonas aeruginosa, Escherichia coli, and Mycobacterium tuberculosis were hosts carrying all four gene categories, highlighting their potential health implications. The neutral community model captured 77.1% and 71.9% of the variance in community dynamics within the conventional and organic farming systems, respectively, indicating that stochastic process was the predominant factor shaping gene communities. The relative smaller m value calculated in organic farming system (0.021 vs 0.023) indicated diminished gene exchange with external sources. Moreover, farming practices were observed to influence the resistance and virulence gene landscape by modifying soil properties, managing heavy metal stress, and steering mobile genetic elements (MGEs) dynamics. The study offers insights that can guide agricultural strategies to address potential health and ecological concerns.

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The life cycle assessment (LCA) methodology currently covers a limited number of human health-related impact categories. Microbiological food safety is an essential aspect for the selection of an appropriate food production system and has been neglected in the LCA so far. A framework for the inclusion of a microbiological food safety indicator, expressed as disability-adjusted life year (DALY) value of the consumed food product to the human health damage category (end-point) was created, and applied in a case study model on the cooked-chilled meals as the ready-to-eat meals can be associated with the occurrence of foodborne illness cases and outbreaks. This study suggests a framework for the inclusion of microbiological risk caused by Bacillus cereus associated with the consumption of ready-to-eat meals (in Belgium) in the LCA. The results indicated that the microbiological risk of one package of the investigated ready-to-eat meal was 1.95 × 10-6 DALY, and the obtained DALY value was included as an impact category in the LCA methodology. Inclusion of other categories of food safety (including chemical safety hazards, pesticide residues, heavy metals, and mycotoxins) in LCA could be done in the same fashion.

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Quantitative microbiological risk assessment (QMRA) methodology aims to estimate and describe the transmission of pathogenic microorganisms from animals and food to humans. In microbiological literature, the availability of whole genome sequencing (WGS) data is rapidly increasing, and incorporating this data into QMRA has the potential to enhance the reliability of risk estimates. This study provides insight into which are the key pathogen properties for incorporating WGS data to enhance risk estimation, through examination of example risk assessments for important foodborne pathogens: Listeria monocytogenes (Lm), Salmonella, Campylobacter and Shiga toxin-producing Escherichia coli. By investigating the relationship between phenotypic pathogen properties and genetic traits, a better understanding was gained regarding their impact on risk assessment. Virulence of Lm was identified as a promising property for associating different symptoms observed in humans with specific genotypes. Data from a genome-wide association study were used to correlate lineages, serotypes, sequence types, clonal complexes and the presence or absence of virulence genes of each strain with patient's symptoms. We also investigated the effect of incorporating WGS data into a QMRA model including relevant genomic traits of Lm, focusing on the dose-response phase of the risk assessment model, as described with the case/exposure ratio. The results highlighted that WGS studies which include phenotypic information must be encouraged, so as to enhance the accuracy of QMRA models. This study also underscores the importance of executing more risk assessments that consider the ongoing advancements in OMICS technologies, thus allowing for a closer investigation of different bacterial subtypes relevant to human health.

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Tenebrio molitor (mealworm) is one of the most promising insect species to produce sustainable feed and food with high nutritional value. Insects may harbour microorganisms both in the gut and on the exoskeleton originating from the rearing environment. Staphylococcus aureus is a pathogenic microorganism frequently involved in food poisoning due to its enterotoxin production. This study aimed to evaluate the S. aureus growth and enterotoxins production following an experimental inoculation into the T. molitor rearing substrate (about 7 log CFU/g). Analyses on the substrate and larvae were performed over a testing period of seven days. The microbial population dynamics were also evaluated through total viable count and lactic acid bacteria count. The effects of fasting, washing, and cooking on the microbial loads of mealworms were evaluated. The results highlighted that mealworms and substrates can maintain their microbial loads of S. aureus over the tested period. Moreover, fasting and washing were generally not able to significantly reduce (p-value > 0.05) S. aureus count in mealworms. On the other hand, cooking significantly reduced (p-value < 0.001) the microbial load in almost all cases. No production of enterotoxins was revealed during the trial. Therefore, microbiological risks can be reduced by a wise choice of substrate, appropriate control measures, and thermal treatment of larvae.

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Foodborne disease caused by nontyphoidal Salmonella (NTS) is one of the most important food safety issues worldwide. The objectives of this study were to carry out microbial monitoring on the prevalence of NTS in commercial ground pork, investigate consumption patterns, and conduct a quantitative microbiological risk assessment (QMRA) that considers cross-contamination to determine the risk caused by consuming ground pork and ready-to-eat food contaminated during food handling in the kitchen in Chengdu, China. The food pathway of ground pork was simplified and assumed to be several units according to the actual situation and our survey data, which were collected from our research or references and substituted into the QMRA model for simulation. The results showed that the prevalence of NTS in ground pork purchased in Chengdu was 69.64% (95% confidence interval [CI], 60.2-78.0), with a mean contamination level of -0.164 log CFU/g. After general cooking, NTS in ground pork could be eliminated (contamination level of zero). The estimated probability of causing salmonellosis per day was 9.43E-06 (95% CI: 8.82E-06-1.00E-05), while the estimated salmonellosis cases per million people per year were 3442 (95% CI: 3218-3666). According to the sensitivity analysis, the occurrence of cross-contamination was the most important factor affecting the probability of salmonellosis. To reduce the risk of salmonellosis caused by NTS through ground pork consumption, reasonable hygiene prevention and control measures should be adopted during food preparation to reduce cross-contamination. This study provides valuable information for household cooking and food safety management in China.

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Ready to Eat (RTE) cooked meat products are among the most consumed RTE food subcategories in the EU/EEA. They are also associated with the highest number of identified listeriosis cases per year (>850), thus posing a public health risk especially among the susceptible population. This study estimated the risk of listeriosis from Italian head cheese (Coppa di Testa) consumption using a Quantitative Microbiological Risk Assessment (QMRA) based on data of prevalence and starting concentrations of Listeria monocytogenes in the product during a 3-year period (n = 1568). A consumer survey (n = 162) was conducted to provide information on domestic storage time and consumption habits, and storage conditions were determined from recordings of temperatures of domestic refrigerators (n = 57). A probabilistic model was designed for the evaluation of the growth of L. monocytogenes at each stage of the product pathway from production to consumption, using Monte Carlo simulations and employing the @Risk software. Risks associated to consumption of vacuum-packed and sliced-at-retail head cheese were assessed: The model predicted that the risk of listeriosis per serving of vacuum-packed product was in the 10-4 and 10-6 range (mean) for the high-risk and general populations respectively, and listeriosis cases were estimated to be greater than those due to consumption of sliced product (with risks in the range of 10-7 and 10-8). Overall, the model predicted that the mean number of listeriosis cases ranged from 0.001 to 0.24 and from 0.06 to 10 per one hundred thousand people, for the healthy and the high-risk population, respectively. Scenario analyses indicated that better control of the temperature of domestic refrigerators is effective in reducing the predicted risk of listeriosis for the longer stored vacuum-packed product by ~80 % for both the healthy and high-risk populations, whereas a shorter use-by-date of 30 days is an effective risk mitigation measure for both types of packed product. Model assumptions, as well as data gaps are discussed.

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Introduction: There is growing interest in the public health and transport sectors in research into exposure to biological hazards, considering not only the risks arising from inter-human contagion, but also those related to exposure to the flight environment itself. The aim of this paper is to report data from an investigation into the water and air-conditioning systems of commercial aircraft for the presence of Legionella contamination, with a total of 645 water samples taken during the period 2007−2021. Methods: The investigation involved 126 aircraft of six different commercial aircraft types: MD80, Airbus A320 F, Embraer 175/190, AIRBUS A330, Boeing 767 and Boeing 777. Water samples were taken from the water systems (toilet taps, galley and boilers). Each sample was preliminarily subjected to an evaluation of the following parameters: temperature, pH and residual chlorine. The ScanVit® Legionella kit was used for bacteria detection and enumeration. Results: Samples were considered positive if the number of colony-forming units/liter (CFU/L) was >100. For the entire observation period, 45% of the investigated aircraft tested positive. Regarding the overall number of samples analyzed, 68.4% (441/645) were below 100 CFU/L, and thus within the limits allowed by the Italian Guidelines. Conclusions: Water system contamination with Legionella in the air transport field is a real public health issue that should not be underestimated given the heavy passenger traffic. Infection should be considered an occupational risk to which crew members are exposed.

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Concerns regarding biological risk in environment have garnered increasing attention. Manure has been believed to be a significant source of antibiotic resistance genes (ARGs) in agricultural soil. Nevertheless, the profile of microbial contamination including ARGs, virulence factor genes (VFGs) and human bacterial pathogens (HBPs) in different manure-amended soils remain largely unknown. Here, we conducted the systematic metagenome-based study to explore changes in resistome, VFGs and HBPs in soils treated by frequently-used manures. The results revealed that many manure-borne ARGs, VFGs, and HBPs could be spreaded into soils, and their diversity and abundance were significantly different among chemical fertilizer, pig manure, chicken manure, cow dung and silkworm excrement application. A total of 157 potential HBPs accounting about 1.33% of total bacteria were detected. The main ARGs transferred from manures to soil conferred resistance to vancomycin and macrolide-lincosamide-streptogramin. The series analysis revealed positive co-occurrence patterns of ARGs-HBPs, VFGs-HBPs and ARGs-VFGs. Microbial contamination were more serious in pig manure and silkworm excrement sample than in the other samples, implying the usage of these two manures increased the risk of HBPs and dissemination of ARGs. This study confirmed the prevalence and discrepancy of resistome, VFGs and HBPs in different manure-amended soils.

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Microbial indicators are often used to monitor microbial safety of aquatic environments. However, information regarding the correlation between microbial indicators and ecotoxicological factors such as potential pathogens and antibiotic resistance genes (ARGs) in anthropogenically impacted waters remains highly limited. Here, we investigated the bacterial community composition, potential pathogens, ARGs diversity, ARG hosts, and horizontal gene transfer (HGT) potential in urban river and wastewater samples from Chaohu Lake Basin using 16S rRNA and metagenomic sequencing. The composition of the microbial community and potential pathogens differed significantly in wastewater and river water samples, and the total relative abundance of fecal indicator bacteria was positively correlated with the total relative abundance of potential pathogens (p < 0.001 and Pearson's r = 0.758). Network analysis indicated that partial ARG subtypes such as dfrE, sul2, and PmrE were significantly correlated with indicator bacteria (p < 0.05 and Pearson's r > 0.6). Notably, Klebsiella was the indicator bacteria significantly correlated with 4 potential pathogens and 14 ARG subtypes. ARGs coexisting with mobile gene elements were mainly found in Thauera, Pseudomonas, Escherichia, and Acinetobacter. Next-generation sequencing (NGS) can be used to conduct preliminary surveys of environmental samples to access potential health risks, thereby facilitating water resources management.

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The objective of this study was to develop a quantitative microbial risk assessment (QMRA) model to evaluate potential risk mitigation strategies to reduce the probability of acquiring hemolytic uremic syndrome (HUS) associated with beef consumption in Argentina. Five scenarios were simulated to evaluate the effect of interventions on the probability of acquiring HUS from Shiga toxin-producing Escherichia coli (STEC)-contaminated ground beef and commercial hamburger consumption. These control strategies were chosen based on previous results of the sensitivity analysis of a baseline QMRA model. The application of improvement actions in abattoirs not applying Hazard Analysis and Critical Control Points (HACCP) for STEC would result 7.6 times lower in the probability that consumers acquired HUS from ground beef consumption, while the implementation of improvements in butcher shops would lead to a smaller reduction. In abattoirs applying HACCP for STEC, the risk of acquiring HUS from commercial hamburger consumption was significantly reduced. Treatment with 2% lactic acid, hot water and irradiation reduced 4.5, 3.5 and 93.1 times the risk of HUS, respectively. The most efficient interventions, in terms of case reduction, being those that are applied in the initial stages of the meat chain.

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Since the late 1990s, the Food and Agriculture Organization (FAO) of the United Nations and the World Health Organization (WHO) has convened expert meetings and consultations to address the microbiological risk assessment (MRA). These meetings are held to provide scientific advice in response to requests for from Codex Alimentarius, the international food standard-setting body. Individuals participate in the FAO/WHO joint expert meetings on the microbiological risk assessment (JEMRA) in their personal capacity, as technical experts, yet bring diverse regional and national perspectives that contribute to practical applications, particularly for low- and middle-income countries (LMICs). Over 370 experts from around the globe have contributed to the meeting outcomes that have been published in nearly 40 monographs in the FAO/WHO microbial risk assessment (MRA) series, addressing particular food commodities with microbial hazard(s) combinations or a methodological aspect of microbial risk assessment. FAO/WHO MRA series inform Codex decision-making for the development of international standards for safe food and faire trade in food products; are consulted by risk managers such as food safety authorities and food business operators to make science-based decisions; and are used by academics to advance food safety research and educate the next generation of food safety professionals.

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OBJECTIVE: A quantitative microbiological risk assessment was conducted for Salmonella spp. in fresh pork from retail stores to consumers in China in order to provide evidence for adopting effective risk management measures. METHODS: The national food safety and risk monitoring data was used to estimate the initial contamination level of Salmonella in the retail fresh pork. The growth model and the cross-contamination model of Salmonella were referenced from the ComBase data base and scientific literature, respectively. Then the human health risk was predicted by the consumption of Salmonella-contaminated fresh pork and the Beta-Poisson dose-response model. RESULTS: The number of Salmonellosis per year among 1 million residents due to fresh pork was estimated to be 4748.The sensitivity analysis showed that the risk of Salmonellosis was mainly and positively correlated with the preparation rate of ready to eat food, the consumption quantity of pork, the initial contamination level of Salmonella in fresh pork, and the proportion of improper cleaning of knives and chopping boards. When the contamination level of Salmonella in fresh pork is controlled below 1 CFU/g, the average probability of illness will be reduced by 59.39%; the average probability of illness will be reduced by 58.97% by increasing the ratio of complete cleaning of chopping boards or the separation ratio of chopping boards for raw and ready-to-eat food to 90%. CONCLUSION: The risk could be effectively reduced by controlling the concentration of Salmonella contamination in retail pork or improving hygiene practices.

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Wastewater treatment plants (WWTPs) are a nonnegligible source of bioaerosols that can pose health risks to workers and nearby residents. Thus, this study systematically investigated the emission characteristics of the size distribution and concentration of Staphylococcus aureus bioaerosol in a WWTP. Then, the research focused on the quantitative microbiological risk assessment (QMRA) of workers and nearby residents for equipping them with various grades personal protective equipment (PPE). Results showed that the peak proportion of the size distributions of bioaerosol particles in the three sources all obtained a size range between 3.3 and 4.7 µm. In the residential building, the peak proportion was larger (>7.0 µm). Referring to the three sources, the average bioaerosol concentrations were in the following sequence: inverted umbrella aerator tank > residual sludge storage yard > microporous aerator tank. The health risks of residents were generally 1-2 orders of magnitude higher than the other two exposure scenarios and were clearly beyond the benchmarks. Meanwhile, the health risks of the field engineer were usually lower than those of the staff at the residual sludge storage yard. In general, equipping workers and residents with PPE could at least decrease the health risks by one order of magnitude, and higher-grade PPE could appropriately promote the reduction of health risks. This research systematically delivered a series of novel data about the emission characteristics of Staphylococcus aureus bioaerosol in a WWTP. It advanced the understanding of the quantitative health risks of equipping individuals with various PPE.

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The inherent differences in microbial behavior among identically treated strains of the same microbial species, referred to as "strain variability", are regarded as an important source of variability in microbiological studies. Biofilms are defined as the structured multicellular communities with complex architecture that enable microorganisms to grow adhered to abiotic or living surfaces and constitute a fundamental aspect of microbial ecology. The research studies assessing the strain variability in biofilm formation are relatively few compared to the ones evaluating other aspects of microbial behavior such as virulence, growth and stress resistance. Among the available research data on intra-species variability in biofilm formation, compiled and discussed in the present review, most of them refer to foodborne pathogens as compared to spoilage microorganisms. Molecular and physiological aspects of biofilm formation potentially related to strain-specific responses, as well as information on the characterization and quantitative description of this type of biological variability are presented and discussed. Despite the considerable amount of available information on the strain variability in biofilm formation, there are certain data gaps and still-existing challenges that future research should cover and address. Current and future advances in systems biology and omics technologies are expected to aid significantly in the explanation of phenotypic strain variability, including biofilm formation variability, allowing for its integration in microbiological risk assessment.

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Food-handling behaviors and risk perceptions among primary food handlers were investigated by consumer surveys from different subjects in 2010 (N = 609; 1st survey will be called here "Year 2010") and 2019 (N = 605; 2nd survey will be called here "Year 2019"). Year 2010 was characterized by consumers' risk perception-behavior gap (i.e., consumers knew safe methods for food-handling, but responses regarding the behaviors did not support their confidence in food safety): they 1) did not wash/trim foods before storage, 2) thawed frozen foods at room temperature, and 3) exposed leftovers to danger zone temperatures. These trends were not improved and the gaps in Year 2010 remained in Year 2019. Year 2010 was also characterized by other common high-risk behaviors improved during 8 years for the following aspects: 1) 70.0% of consumers divided a large portion of food into smaller pieces for storage, but few consumers (12.5%) labeled divided foods with relevant information, and 2) they excessively reused kitchen utensils. Whereas in Year 2019, more consumers (25.7%) labeled food and usage periods for kitchen utensils were shortened. Consumers usually conformed to food safety rules in both Year 2010 and 2019: 1) separate storage of foods, 2) storage of foods in the proper places/periods, 3) washing fruits/vegetables before eating, 4) washing hands after handling potentially hazardous foods, and 5) cooking foods and reheating leftovers to eat. Our findings provided resources for understanding consumers' high-risk behaviors/perceptions at home, highlighting the importance of behavioral control.

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The main results and prospects of fundamental and applied hygienic research of the laboratory of biosafety and nutrimicrobiome analysis of the Federal Research Centre of Nutrition, Biotechnology and Food Safety (hereinafter - the Institute of Nutrition) in the direction of developing a regulatory and methodological framework for assessing the microbiological safety of food are reviewed. The formation of microbiological regulation as a scientific analytical and administrative managerial process in the former USSR and the Russian Federation is considered in the context of historical data, including personal contribution of the scientists of the Institute of Nutrition and other specialists. The basic principles of regulation are emphasized: the scientific validity of the established criteria and requirements, the feasibility, technological attainability, differentiation according to the degree of danger to the health of consumers, preventive nature. The resource of the national normative and methodological base in the field of microbiological food safety at the turn of the century is characterized, the features of the introduction of the microbiological risk assessment (MRA) methodology in the substantiation of Russian norms and measures for the prevention of food infections are described. The information is given on the developed guidance documents on MRA and on the examples of norms adopted on its basis. The article covers the issues of reglamentation the requirements for food safety and reducing the spread of new pathogens Stx-Escherichia coli, Listeria monocytogenes, Enterobacter sakazakii, Campylobacter spp. in the food chain based on risk-oriented approaches. The necessity of taking specific measures for the prevention of cross-contamination in the poultry processing industry is substantiated, taking into account the evidence of the high adaptability of C. jejuni isolated from domestic raw poultry. In the sanitarian-mycological aspect, the monitoring perspective of mould fungi, taking into account their chemotypes, in cereals and non-grain plant products is shown to predict the risk of mycotoxin accumulation and take timely measures. The need to assess the impact on the population, taking into account the characteristics of consumption in the country, as well as the development of criteria for indirect risk of residues are argued for regulation of the antibiotics in food. In light of the challenges in the field of agro- and food technologies to public health at the present stage, contributing to the acceleration of microbial evolution and the emergence of new risks in food, the priority tasks of improving the regulatory and methodological base for assessing microbiological safety have been identified, with an emphasis on the introduction into the process of substantiating the norms of innovative OMICs-technologies based on the achievements of genomics, transcriptomics, proteomics, metabolomics, bioinformatics.

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Although consumers' food purchase/transport have been reported as causes of food safety risks, there is a lack of empirical data that are feasible to identify persistent and emerging risky behaviors of consumers. This longitudinal trend study consists of individual consumer surveys in 2010 (n = 609) and 2019 (n = 605) to analyze changes in risky behaviors linked to food purchase/transport over a decade. Overall, the results identified purchase/transport time and purchase order as the emerging and unchanged risk factors, respectively. Consumers' preferences into channels for purchase (large discount stores rather than small/traditional markets) and transport (using cars or delivery) implied the convenience as the noticeable trend. Whereas, unexpected increases in purchase/transport time highlighted the underestimated risks in long-term exposure of foods under inadequate temperature. Food should not be exposed to danger zones > 1-2 h, but consumers might be unaware of the risk especially for preferred channels (e.g., 77 and 36 min. are required for purchase and transport from large discount stores, respectively). In the case of unchanged risky behavior, more than half of consumers in both surveys did not follow proper purchasing orders. Our findings highlight the necessity for novel countermeasures and the improvement of current consumer guidelines against emerging and unchanged risky behaviors, respectively.

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