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In this study, the distribution of hygienic indicator bacteria in cattle livers and bile was examined at slaughterhouses. One hundred twenty-seven cattle livers with gallbladders were carefully eviscerated from carcasses at 10 slaughterhouses. Microbiological examination revealed that nine bile samples (7.1% prevalence) and 19 liver parenchyma samples (15.0% prevalence) were positive for Enterobacteriaceae (EB) with means ± standard deviations of 3.68 ± 4.63 log CFU/mL and 1.59 ± 2.47 log CFU/g, respectively; thus, bacterial contamination was apparent even at the postevisceration stage. Subsequently, 70 cattle livers were obtained at the postprocessing and storage stage from 7 of the 10 slaughterhouses. Microbiological analysis revealed significantly higher levels of EB in the liver parenchyma (3.00 ± 3.89 log CFU/g, P = 0.011) than those at the postevisceration stage, suggesting that bacterial dissemination and/or replication occurred in the liver parenchyma during processing and storage. According to 16S rRNA ion semiconductor sequencing analysis of representative samples from 12 cattle, Proteobacteria, Firmicutes, and Actinobacteria were dominant in both the parenchyma and bile in which EB and Escherichia coli were predominant among livers with higher EB levels. These results suggest that bile plays a role as a vehicle for bacterial transmission to the liver parenchyma. This study is the first to evaluate bacterial distribution and community structure in the liver and biliary microecosystem of cattle at slaughter. Our data support the use of EB testing of bile to screen cattle livers contaminated with high levels of fecal indicator bacteria.

Cheese made with unpasteurized milk has been associated with outbreaks of illness. However, there are limited data on the prevalence of Shiga toxin-producing Escherichia coli (STEC) in these products and a lack of clarity over the significance of E. coli as a general indicator of hygiene in raw milk cheeses. The aim of this study was to provide further data to address both of these issues, as well as assessing the overall microbiological quality of raw milk cheeses available to consumers in England. A total of 629 samples of cheese were collected from retailers, catering premises, and manufacturers throughout England. The majority (80%) were made using cow's milk, with 14% made from sheep's milk and 5% from goat's milk. Samples were from 18 different countries of origin, with the majority originating from either the United Kingdom (40%) or France (35%). When interpreted against European Union microbiological criteria and United Kingdom guidance, 82% were considered to be of satisfactory microbiological quality, 5% were borderline, and 12% were unsatisfactory. Four samples (0.6%) were potentially injurious to health due to the isolation of STEC from one,104 CFU/g of coagulase-positive staphylococci in two, and100 CFU/g of Listeria monocytogenes in the fourth sample. Indicator E. coli and Listeria species were detected more frequently in soft compared with hard cheese. Higher levels of indicator E. coli were significantly associated with a greater likelihood of detecting Shiga toxin genes (stx1 and/or stx2).

Although many neurotoxicity prediction studies of food additives have been developed, they are applicable in a qualitative way. We aimed to develop a novel prediction score that is described quantitatively and precisely. We examined cell viability, reactive oxygen species activity, intracellular calcium and RNA transcription level of potential prediction related genes to develop a high-throughput neurotoxicity test method in vitro to screen the neurotoxicity of hazardous factors in food using AI-based machine learning. We trained artificial intelligence models (random forest and neural network) to predict neurotoxicity precisely, establishing a universal classification assessment score (CA-Score) that relies on the expression status of only 13 of prediction related genes. The CA-Score system is almost universally applicable to food risk factors (p

Outbreaks of food poisoning due to the consumption of norovirus-contaminated shellfish continue to occur. Male-specific (F+) coliphage has been suggested as an indicator of viral species due to the association with animal and human wastes. Here, we compared two methods, the double agar overlay and the quantitative real-time PCR (RT-PCR)-based method, for evaluating the applicability of F+ coliphage-based detection technique in microbial contamination tracking of shellfish samples. The RT-PCR-based method showed 1.6-39 times higher coliphage PFU values from spiked shellfish samples, in relation to the double agar overlay method. These differences indicated that the RT-PCR-based technique can detect both intact viruses and non-particle-protected viral DNA/RNA, suggesting that the RT-PCR based method could be a more efficient tool for tracking microbial contamination in shellfish. However, the virome information on F+ coliphage-contaminated oyster samples revealed that the high specificity of the RT-PCR- based method has a limitation in microbial contamination tracking due to the genomic diversity of F+ coliphages. Further research on the development of appropriate primer sets for microbial contamination tracking is therefore necessary. This study provides preliminary insight that should be examined in the search for suitable microbial contamination tracking methods to control the sanitation of shellfish and related seawater.

Fresh produce continues to be the main source of foodborne illness outbreaks in the United States, implicating bacterial pathogens such as Escherichia coli O157:H7 (EHEC). The efficacy of nanoemulsified carvacrol (NCR) as a washing treatment in reducing EHEC on fresh produce was investigated. Fresh baby spinach, romaine lettuce, and iceberg lettuce leaves (2.5-cm-diameter cores) were spot inoculated with a five-strain cocktail of nalidixic acid-resistant EHEC at ∼6 log CFU/cm2. After air drying for 1 h, 20 pieces of each inoculated produce leaf were immersed in water-based treatment solutions (200 mL per group), including water alone, 25 or 50 ppm of free chlorine, and 0.25 or 0.75% NCR for 2 min. Inoculated produce leaves without any treatment served as baseline. Produce leaves were stored at 10°C, and surviving EHEC populations were enumerated on days 0, 2, 7, and 14. The viability of EHEC following NCR treatments on the fresh produce was visualized under a fluorescence microscope. NCR treatment at 0.75% immediately reduced EHEC populations on iceberg lettuce by 1.3 log CFU/cm2 as compared with the produce treated with water alone (P0.05). Antimicrobial activity of NCR against EHEC was comparable to chlorine treatments on day 0 for all produce (P0.05). After 14 days of storage at 10°C, populations of EHEC on 0.75% NCR-treated romaine lettuce were reduced by 2.3 log CFU/cm2 compared with the recovery from 50 ppm of chlorine-treated samples (P0.05). Microscopic images revealed that EHEC cells were observed to be clustered on the baseline samples, indicating the development of cell aggregation, compared with the scattered cells seen on NCR-treated leaf surfaces. Treatments with NCR did not significantly affect the color of the fresh produce leaves during 14 days of storage at 10°C. Results of this study support the potential use of NCR as a water-soluble natural antimicrobial wash treatment for controlling EHEC on fresh produce.

The present study investigated the heavy metal (HM) accumulation in some food items and associated potential human health risk in the Prakasam district. Food samples were collected from 9 villages of different mandals mainly comprising of rice, wheat, green gram, red gram, black gram, potato, and amaranthus. The HMs chromium (Cr), arsenic (As), lead (Pb), cadmium (Cd), and mercury (Hg) were analyzed by Inductively Coupled Plasma Mass Spectroscopy (ICPMS). The results showed that the HM concentrations in vegetables were higher than the cereals and pulses. The decreasing order of HM concentrations were observed as Cr > As > Pb > Cd > Hg, Cr > Pb > As = Cd > Hg and Cr > Pb > Cd > As > Hg in cereals, pulses, and vegetables, respectively. The dietary intake of cereals and vegetables contributed more exposure to HMs. The hazard index (HI) in case of cereals (3.648) and vegetables (1.003) exceeded 1 indicating the non-carcinogenic risk when these food items are consumed by adults. The HI in case of cereals (7.106) and vegetables (1.747) exceeded 1 indicating the health risks when these food items were consumed by children. The HI for cereals and vegetables in case of children is almost double to the HI of adults. The lifetime cancer risk was observed for Cr via consumption of rice and amaranthus, both in adults and children. In conclusion, health risk due to consumption of cereals and vegetables revealed both non-carcinogenic and carcinogenic risk for adults and children. The findings of the present study can be used for the development of potential strategies for risk control and management.

The concentration of meHg in freshwater fish and seafood was investigated, as well as the consumption patterns of fish and seafood by different demographic groups (age, ethnicity, gender). A potential alarm for human health hazards was also assessed, and the results were compared to the provisional tolerable weekly intakes (PTWIs) and the hazard quotient parameter (HQ). The results showed that meHg levels of 67 species ranged from 0.013 to 0.252 mg/kg of wet weight (WW) with significant differences between different fish and seafood groups (χ2KW = 49.09; p pelagic fish (0.0686 mg/kg WW) > freshwater fish 0.045 mg/kg WW) > cephalopods (0.0405 mg/kg WW) crustaceans (0.0356 mg/kg WW). The results revealed that older population (> 40 years old) consumed significantly (p = 0.000) more fish compared to younger generations and the elderly consumed the highest amounts of fish (104.0 ± 113.0 g/day). The adolescents (10–17 years old) consumed more than double of amount for both cephalopod and crustacean compared to the older populations (p p = 0.000) higher amounts of fish and seafood compared to other ethnicities, similar to male subjects (95.2 ± 102 g/day; p = 0.026) when compared to the female (86 ± 96.3 g/day). The estimated weekly intake (EWI) values showed results below 1.6 µg/kg BW/week, the tolerable levels recommended by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) for all different demographic factors except for higher consumers at 75th percentile and above. Consumption of marine fish contributed to a higher value of PTWI to all different demographic groups (the estimated weekly intake (EWI) range: 0.2988–0.6893 µg/kg BW/week) but for the adolescents, where from the consumption of crustaceans (0.3488 µg/kg BW/week or 21.8% of PTWI) and cephalopods (0.504 µg/kg BW/week or 31.5% of PTWI). The results from this study also revealed the HQ value for overall consumption of fish and seafood by the adolescents and elderly exceeded one. This was contributed from the consumption of demersal fish and cephalopods, thus indicating the nonacceptable level of noncarcinogenic adverse health effects.

Consumption advice to ensure the health and safety of fish consumers remains urgent to handle the ever-increasing panic over heavy metal toxicity. Moreover, studies of fish consumption rarely focus on the perceptions and awareness of consumers. Considering this, the present study examines the factors determining the willingness to follow fish consumption advice as well as calculate the risk–benefit ratio and produce the vulnerability map followed by formulating fish consumption advice for consumers of two commercially important fish species, tilapia (Oreochromis niloticus) and mudfish (Ophiocephalus striatus), in the Laguna de Bay area of the Philippines. Primary data on consumers’ perceptions were collected through a questionnaire, whereas heavy metal contamination data were compiled from the best available literature. We concluded that people’s willingness to adopt consumption advice is mostly dependent on their existing level of fish consumption (P = 0.000). There was no discernible difference in the mean concentration of heavy metals in fish between the dry and wet seasons, with the exception of As concentrations in the samples (t15.75 = 3.72, p

Ochratoxins (OTs) with nephrotoxic, immunosuppressive, teratogenic, and carcinogenic properties are thermostable fungal subordinate metabolites. OTs contamination can occur before or after harvesting, during the processing, packing, distribution, and storage of food. Mold development and mycotoxin contamination can occur in any crop or cereal that has not been stored properly for long periods of time and is subjected to high levels of humidity and temperature. Ochratoxin A (OTA) presents a significant health threat to creatures and individuals. There is also a concern of how human interaction with OTA will also express the remains of OTA from feedstuffs into animal-derived items. Numerous approaches have been studied for the reduction of the OTA content in agronomic products. These methods can be classified into two major classes: inhibition of OTA adulteration and decontamination or detoxification of food. A description of the various mycotoxins, the organism responsible for the development of mycotoxins, and their adverse effects are given. In the current paper, the incidence of OTA in various fodder and food materials is discussed, which is accompanied by a brief overview of the OTA mode of synthesis, physicochemical properties, toxic effects of various types of ochratoxins, and OTA decontamination adaptation methods. To our knowledge, we are the first to report on the structure of many naturally accessible OTAs and OTA metabolism. Finally, this paper seeks to be insightful and draw attention to dangerous OTA, which is too frequently neglected and overlooked in farm duplication from the list of discrepancy studies.

The study vigilantly considered the load of Pb, Cd, Cr, Cu, and Zn in a variety of tissues (muscle, gills, and liver) of 5 fish species (Mystus gulio, Notopterus notopterus, Notopterus chitala, Mugil cephalus, and Glossogobius giuris) collected from six sites in the lower Gangetic area. The study showed the lowest concentration of metals in the muscles. The accumulated patterns of heavy metals differed in different regions and concentrations fluctuated between the liver and gills. The target hazard quotient (THQ) value has been measured in contaminated fish. The THQ values for all the metals in respective fishes are below 1 that indicate that indirect intake of metals by consuming these selected fishes will not result in potential health hazard in human. The estimated daily intake (EDI) results were also calculated. EDI levels of all elements are lower than the permissible limit indicating a lower chance for health risk to occur. However, doses below the recommended levels do not indicate that they are completely safe for consumption or those above are not to be used. Thus, it can be demonstrated that occurrence of Cd, Cr, Pb, Cu, and Zn in the preferred tissues of the selected fish species in the present study may not pretense severe human health risk after consumption at its existing concentration.

Milk protein is recognised as one of the eight most common food allergens in the United States, and one of the most common causes of fatal or near-fatal food-induced anaphylactic reactions globally. Undeclared allergens in food products are the leading cause of the U.S. Food and Drug Administration (FDA) requests for food recalls, with undeclared milk protein as the most frequently cited allergen. The purpose of this study was to evaluate the prevalence of milk protein contamination in milk-alternative ice cream products sold as frozen dessert in the U.S. and labelled as 'non-dairy', 'dairy-free', and/or 'vegan', and to characterise the potential risk for allergic individuals consuming these products. Thirty-two unique milk-alternative frozen dessert products, including flavours with and without inclusions, from 16 different brands, were tested for the presence of milk proteins using enzyme-linked immunosorbent assay. Milk protein was detected in three of the tested products: two contained solid inclusions, and one did not have inclusions. One of the three products with detectable milk protein had an advisory statement indicating the product was made in a facility that processed milk. Although no consensus exists as to the minimal dose of milk protein necessary to elicit an adverse effect in a milk-sensitised individual, the milk protein quantities in one serving of each of the three products exceeded some of the published thresholds. These results suggest that one serving of certain milk-alternative frozen dessert products could potentially elicit an allergic response in highly sensitive individuals.

The ubiquity of Listeria monocytogenes in the environment affects the food industry and presents concerns for frozen food facilities. This study determined the prevalence and numbers of Listeria species and L. monocytogenes on raw produce arriving at frozen food facilities. Raw produce was collected using multilevel blinding protocols to ensure anonymity of participants and avoid traceback. Five raw vegetables were selected: corn, carrots, green beans, peas, and spinach. Raw products were collected after arrival at the facilities but before cleaning or other preprocessing steps that are typically performed inside the facility. The U.S. Food and Drug Administration's Bacteriological Analytical Manual method for detection of Listeria spp. and L. monocytogenes was followed, with PCR screening followed by selective plating methods. Listeria numbers were estimated from positive samples using the most-probable-number (MPN) methodology. A total of 290 samples were collected, with 96 and 17 samples positive for Listeria spp. (33.1%) and L. monocytogenes (5.9%), respectively. Enumeration data for the 96 Listeria spp. samples indicated 82 samples had greater than 100 MPN of Listeria spp. per g and 14 samples had less than 100 MPN Listeria spp. per g. The prevalence of Listeria spp. varied by commodity: spinach (66.7%), peas (50%), corn (32.2%), green beans (22.2%), and carrots (13%). L. monocytogenes prevalence was determined in corn (13.6%), peas (6.3%), and green beans (4.2%) arriving at processing facilities. Such data were previously unavailable to frozen vegetable processors and are valuable in implementing process control standards. The prevalence and pathogen concentration data from raw commodities found in this study can provide the industry with information to conduct more accurate quantitative risk assessments and a baseline to model and target appropriate pathogen reduction steps during processing. HIGHLIGHTS

Food contamination caused by foodborne pathogens is one of the most significant concerns in public health worldwide, and accounts for a significant portion of food loss every year. The emergence of antimicrobial resistant bacteria has turned the researchers' attention back to the potential of bacteriophages as antibacterial agents, which have been attempted in various pre-and post-harvest food production settings. While the application of phage-based antibacterial products has achieved considerable success, a number of technical, environmental and administrative challenges remain unaddressed. In this review, we summarized the current status of bacteriophage application in food industry. More importantly, we discussed the obstacles facing the further development of phage-based antibacterial products from the aspects of technology, environmental safety, and administrative policy. We also put forward some possible solutions to these challenges, serving as reference information for future studies. This article is protected by copyright. All rights reserved.

An improved fish smoking oven called FAO-Thiaroye Technique (FTT) has been introduced in Ghana and other countries in the Global South as a technical intervention for the high levels of polycyclic aromatic hydrocarbons (PAHs) in traditionally smoked fish produced in those regions. This study evaluated the extent to which the intervention reduces consumer exposure to PAHs (considering benzo(a)pyrene [BaP] as a marker) in smoked fish, using Ghana as a case. Smoked Sardinella sp. were sampled from two traditional ovens (Chorkor smoker and metal drum oven) and the FTT and their PAH levels were determined by gas chromatography-mass spectrometry. Samples of the product were also purchased from informal markets in three selected regions of Ghana and analyzed for their PAH levels. Cross-sectional consumer surveys were conducted in the selected regions to determine intakes of the commodity. A probabilistic risk assessment of PAH was then done by the margin of exposure (MoE) approach. BaP MoE as low as 1,060 and 752 were obtained for products from the traditional ovens and the informal markets, respectively, whereas the lowest value for FTT products was approximately 161,000. MoE values less than 10,000 were considered to denote a serious public health concern requiring risk management action. Therefore, the findings suggest that there is a potential health concern of high consumer exposure to PAHs in traditionally smoked fish in Ghana, and that the FTT is a technically viable intervention for the problem.

In this study, the residue data for conazole fungicides were collated and analyzed in all crop samples reported by the United States Department of Agriculture Pesticide Data Program over the period of 2009-2019. Considering all individual samples, the overall detection frequencies (DFs) of conazoles are less than 13%. Among the 18 conazoles, imazalil had the highest overall DF of 6%, followed by tebuconazole and myclobutanil, with 4% each. Conazoles were detected most frequently in raisins with 28% DF, followed by cherries (frozen and fresh) and grapes, with 12, 10, and 10%, respectively. The presence of multiple conazoles in single commodity samples is very low, below 2%. The analyses found no more than four conazoles present in any given sample. Out of the 18 conazoles, 8 of them were not detected in more than 99.9% of the commodity samples from 2009 to 2019 and, therefore, can be eliminated from screening-level cumulative risk assessment for dietary contributions from food items. While conazoles are widely used on food commodities, co-occurrence of conazole residues was observed only in a very limited number of food commodities, including raisins, grapes, cherries (frozen), nectarines, and peaches. Considering the remaining individual food commodities, the co-occurrence of conazole residues in single commodity samples is very low or not even present.

Listeria monocytogenes is a saprophytic gram-positive bacterium, and an opportunistic foodborne pathogen that can produce listeriosis in humans and animals. It has evolved an exceptional ability to adapt to stress conditions encountered in different environments, resulting in a ubiquitous distribution. Because some food preservation methods and disinfection protocols in food-processing environments cannot efficiently prevent contaminations, L. monocytogenes constitutes a threat to human health and a challenge to food safety. In the host, Listeria colonizes the gastrointestinal tract, crosses the intestinal barrier, and disseminates through the blood to target organs. In immunocompromised individuals, the elderly, and pregnant women, the pathogen can cross the blood-brain and placental barriers, leading to neurolisteriosis and materno-fetal listeriosis. Molecular and cell biology studies of infection have proven L. monocytogenes to be a versatile pathogen that deploys unique strategies to invade different cell types, survive and move inside the eukaryotic host cell, and spread from cell to cell. Here, we present the multifaceted Listeria life cycle from a comprehensive perspective. We discuss genetic features of pathogenic Listeria species, analyze factors involved in food contamination, and review bacterial strategies to tolerate stresses encountered both during food processing and along the host’s gastrointestinal tract. Then we dissect host–pathogen interactions underlying listerial pathogenesis in mammals from a cell biology and systemic point of view. Finally, we summarize the epidemiology, pathophysiology, and clinical features of listeriosis in humans and animals. This work aims to gather information from different fields crucial for a comprehensive understanding of the pathogenesis of L. monocytogenes., Graphical abstract

Listeria monocytogenes is a ubiquitous pathogen that can cause morbidity and mortality in immunocompromised individuals. Growth of L. monocytogenes is possible at refrigeration temperatures due to its psychrotrophic nature. The use of antimicrobials in dairy products is a potential way to control L. monocytogenes growth in processes with no thermal kill step, thereby enhancing the safety of such products. Microbial-based enzymes offer a clean-label approach for control of L. monocytogenes outgrowth. Lactose oxidase (LO) is a microbial-derived enzyme with antimicrobial properties. It oxidizes lactose into lactobionic acid and reduces oxygen, generating H2O2. This study investigated the effects of LO in UHT skim milk using different L. monocytogenes contamination scenarios. These LO treatments were then applied to raw milk with various modifications; higher levels of LO as well as supplementation with thiocyanate were added to activate the lactoperoxidase system, a natural antimicrobial system present in milk. In UHT skim milk, concentrations of 0.0060, 0.012, and 0.12 g/L LO each reduced L. monocytogenes counts to below the limit of detection between 14 and 21 d of refrigerated storage, dependent on the concentration of LO. In the 48-h trials in UHT skim milk, LO treatments were effective in a concentration-dependent fashion. The highest concentration of LO in the 21-d trials, 0.12 g/L, did not show great inhibition over 48 h, so concentrations were increased for these experiments. In the lower inoculum, after 48 h, a 12 g/L LO treatment reached levels of 1.7 log cfu/mL, a reduction of 1.3 log cfu/mL from the initial inoculum, whereas the control grew out to approximately 4 log cfu/mL, an increase of 1 log cfu/mL from the inoculum on d 0. When a higher challenge inoculum of 5 log cfu/mL was used, the 0.12 g/L and 1.2 g/L treatments reduced the levels by 0.2 to 0.3 log cfu/mL below the initial inoculum and the 12 g/L treatment by >1 log cfu/mL below the initial inoculum by hour 48 of storage at refrigeration temperatures. After the efficacy of LO was determined in UHT skim milk, LO treatments were applied to raw milk. Concentrations of LO were increased, and the addition of thiocyanate was investigated to supplement the effect of the lactoperoxidase system against L. monocytogenes. When raw milk was inoculated with 2 log cfu/mL, 1.2 g/L LO alone and combined with sodium thiocyanate reduced ~0.8 log cfu/mL from the initial inoculum on d 7 of storage, whereas the control grew out to >1 log cfu/mL from the initial inoculum. Furthermore, in the higher inoculum, 1.2 g/L LO combined with sodium thiocyanate reduced L. monocytogenes counts from the initial inoculum by >1 log cfu/mL, whereas the control grew out 2 log cfu/mL from the initial inoculum. Results from this study suggest that LO is inhibitory against L. monocytogenes in UHT skim milk and in raw milk. Therefore, LO may be an effective treatment to prevent L. monocytogenes outgrowth, increase the safety of raw milk, and be used as an effective agent to prevent L. monocytogenes proliferation in fresh cheese and other dairy products. This enzymatic approach is a novel application to control the foodborne pathogen L. monocytogenes in dairy products.

The first step in quantitative microbial risk assessment (QMRA) is to determine the distribution of pathogen contamination among servings of the food in question at some point in the farm-to-table chain. In the present study, the distribution of Salmonella contamination among servings of chicken liver for use in the QMRA was determined at meal preparation. Salmonella prevalence (P), most probable number (MPN, N), and serotype for different serving sizes were determined by use of a combination of five methods: (i) whole sample enrichment; (ii) quantitative PCR; (iii) culture isolation; (iv) serotyping; and (v) Monte Carlo simulation. Epidemiological data also were used to convert serotype data to virulence (V) values for use in the QMRA. A Monte Carlo simulation model based in Excel and simulated with @Risk predicted Salmonella P, N, serotype, and V as a function of a serving size of one (58 g) to eight (464 g) chicken livers. Salmonella P of chicken livers was 72.5% (58 of 80) per 58 g. Four Salmonella serotypes were isolated from chicken livers: (i) Infantis (P = 28%, V = 4.5); (ii) Enteritidis (P = 15%, V = 5); (iii) Typhimurium (P = 15%, V = 4.8); and (iv) Kentucky (P = 15%, V = 0.8). Salmonella N was 1.76 log MPN/58 g (median) with a range of 0 to 4.67 log MPN/58 g, and the median Salmonella N was not affected (P > 0.05) by serotype. The model predicted a nonlinear increase (P ≤ 0.05) of Salmonella P from 72.5%/58 g to 100%/464 g, a minimum N of 0 log MPN/58 g to 1.28 log MPN/464 g, and a median N from 1.76 log MPN/58 g to 3.22 log MPN/464 g. Regardless of serving size, predicted maximum N was 4.74 log MPN per serving, mean V was 3.9 per serving, and total N was 6.65 log MPN per lot (10,000 chicken livers). The data acquired and modeled in this study address an important data gap in the QMRA for Salmonella and whole chicken liver. HIGHLIGHTS

The United States is one of the largest catfish producers in the world. Louisiana is the leading producer of wild-caught catfish. Historically, the U.S. Food and Drug Administration inspected all seafood products; however, in 2008, Congress moved the inspection of fish in the order Siluriformes to the U.S. Department of Agriculture (USDA), Food Safety and Inspection Service. Full enforcement of the rule began on 1 September 2017. The present study was conducted to assess the impact of USDA Siluriformes fish regulation on small Louisiana wild-caught catfish processors and to determine the microbiological quality of and Salmonella prevalence in raw fillets. Nine facilities participated in the assessment study. Surveys were conducted before and after full enforcement to identify whether facilities had established prerequisite programs and record keeping associated with sanitation, hazard analysis and critical control point (HACCP) plans, food defense, and product recall. The processors' attitude about the change in regulations also was analyzed. For analysis of the microbiological quality and Salmonella prevalence, catfish samples were collected once per month for 2 years. Samples were evaluated for aerobic bacteria counts (APC), coliforms, Escherichia coli, Staphylococcus aureus, and Salmonella. The preenforcement survey revealed that only one facility had developed a HACCP plan, but it was not implemented. After 1 year of full enforcement, all the facilities developed and implemented a HACCP plan to process fresh catfish, and 78% of the processors reported a reduction in the amount of catfish processed due to limits in hours of operation and loss of fishermen. For microbiological quality, the mean (±SD) APC and counts of E. coli, coliforms, and S. aureus were 5.01 ± 0.70, 0.58 ± 0.89, 2.16 ± 0.77, and 0.73 ± 1.02 log CFU/g, respectively; 5.3% of the samples was confirmed positive for Salmonella. These findings indicate that after USDA enforcement, facilities improved food safety program documentation; however, the processing practices did not change. The microbial quality of the catfish fillets was within the acceptable levels in accordance with the International Commission on Microbiological Specifications for Foods. HIGHLIGHTS

AIMS We investigated the fate of Salmonella in lettuce seeds grown in a hydroponic system and the potentials of applying photodynamic inactivation (PDI) to enhance microbial safety of hydroponic farming systems. METHODS AND RESULTS Lettuce was grown from Salmonella-contaminated seeds, and rose bengal-mediated PDI was applied. Without intervention, Salmonella could persist in plants and hydroponic farming environment throughout 6 weeks of lettuce growth. Cross-contamination from Salmonella-inoculated to noninoculated seedlings was observed. PDI significantly decreased Salmonella from 3.90 ± 0.31 log colony-forming unit (CFU) per plant to 2.77 ± 0.49 log CFU per plant without extra illumination needed (p

Edible insects are a novel food in most countries; their popularity is growing because of their high protein/low fat content, ease of cultivation, and small environmental impact. To our knowledge, this is the first report that addresses both microbiological and chemical hazards in edible insects. Samples were collected from retail stores or purchase through e-commerce. A total of 51 samples of dried whole insects or insect powders were tested for generic Escherichia coli ( E. coli ) which serves as an indicator of the overall sanitation conditions throughout the food production chain, and the bacterial pathogen Salmonella species (spp.) Neither Salmonella spp. nor generic E. coli (>100 Colony Forming Units (CFU)/g) were found in any of the samples. 43 samples of crickets (protein bars, powders, flours, whole insects) and 4 samples of silkworm (whole insects) were analyzed for up to 511 pesticides. 39 samples contained residues from one to four pesticides; 34 samples were compliant and 5 samples were non-compliant with Canadian regulations. Seven different pesticide residues were detected with glyphosate and its metabolite, AMPA, being the predominant residues detected. Nineteen of the samples tested for pesticides were also analyzed for arsenic, cadmium, mercury, and lead; there was insufficient material remaining to allow for metals testing. The positive rate for arsenic, cadmium, lead and mercury was 100%, 79%, 58% and 74%, respectively. The detected concentrations ranged from 0.030 mg/kg to 0.34 mg/kg for arsenic, from 0.031 mg/kg to 0.23 mg/kg for cadmium, 0.019 mg/kg to 0.059 mg/kg for lead, and from 0.00094 mg/kg to 0.028 mg/kg for mercury. Based on the lack of detection of microbiological contamination, and the positive rate and levels of pesticides and metals observed in the products, Health Canada determined that all insect products analyzed were safe for human consumption. This is a limited study; CFIA will continue to monitor this novel food.

Science-based guidance was used at eight small and very small state and federally inspected ready-to-eat meat and poultry processors across Michigan. Data were collected to determine the current level of sanitary control methods used for reducing Listeria in the processing environment and compared interactions with the facility microbial results. A checklist was created to assess the current recommended sanitary control methods from the U.S. Department of Agriculture, U.S. Food and Drug Administration, and the Michigan Department of Agriculture and Rural Development. The checklist, composed of 178 items divided into 10 general content domains, was used to assess which of the recommended controls were being used in the facilities to prevent postlethality contamination of ready-to-eat products. Effectiveness of preoperational and operational sanitation was assessed through sampling 12 nonfood contact surfaces by using an ATP reader and amplified nucleic single temperature reaction test for Listeria spp., including L. monocytogenes, at each facility. In total, 288 samples were taken collectively from the eight facilities (96 ATP, 96 preoperational Listeria spp., and 96 operational Listeria spp.). Microbial outcomes did not differ (P > 0.05) based on the overall number of recommended sanitary control methods used and the type of facility inspection. There was a greater content domain compliance overall in operational sanitation (P = 0.0005), sanitation (P = 0.0030), facility (P = 0.0397), and personal hygiene (P = 0.0033) than for segregation procedures regardless of the regulating body. Findings suggest that regardless of the regulating body, the quality of sanitary control measures used is more impactful for microbial control than simply the quantity implemented. Pathogen control may be obtained without implementing all of the sanitary control methods within the guidance documents. HIGHLIGHTS

Fungi in the Fusarium genus produce trichothecene mycotoxins including deoxynivalenol (DON) and T-2 toxin which may elicit their damaging effects on the gastrointestinal tract following the consumption of contaminated cereal-based foods. The aim of our study was to evaluate the effects of these commonly occurring fusarotoxins alone and in combination using the human, non-cancerous intestinal epithelial cell line HIEC-6. Based on our experimental data, 24 h after treatment with fusarotoxins, hydrogen peroxide levels, intracellular oxidative stress and the amounts of inflammatory interleukins IL-6 and IL-8 significantly increased. Cell membrane localization of the tight junction protein claudin-1 decreased, whereas distribution of occludin remained unchanged. Taken together, the HIEC-6 cell line appears to be a suitable experimental model for monitoring the combined effects of mycotoxins at the cellular level including changes in the redox states of cells.

AIMS To gather data on agricultural practices in organic farms in Sao Paulo, Brazil, and evaluate their relationship with the microbiological characteristics of samples collected along the production chain. METHODS AND RESULTS Practices data were based on field observations and interviews with farmers in 10 selected organic lettuce producing farms. Counts of Enterobacteriaceae and surveys for Salmonella were performed in samples of lettuce (before and after washing), fertilizers, irrigation and washing water, all collected in the same farm. Water samples were also tested for total coliforms and generic Escherichia coli. Isolated Enterobacteriaceae were identified by MALDI-TOF MS. Contamination of lettuce was influenced by some agricultural practices: chicken manure-based fertilization resulted in higher Enterobacteriaceae counts in lettuce when compared to other types of manure, whereas pre-washed lettuces presented lower microbial counts than non-pre-washed samples. Salmonella was detected in one lettuce sample by qPCR. Escherichia coli was detected in all irrigation water samples. All sample types contained Enterobacteriaceae species commonly reported as opportunistic human pathogens. CONCLUSIONS The data highlight the need for improvement in the good agricultural practices in the studied farms. SIGNIFICANCE AND IMPACT OF THE STUDY This study provides information on agricultural practices and microbiological characteristics of organic lettuce, contributing to the development of more accurate risk assessments.

Although postharvest coffee fruit fermentation can improve coffee flavour and quality, the mycotoxin ochratoxin A (OTA) can also be a result of microbiological activity, albeit in the later drying step of coffee processing. To evaluate the possible occurrence of OTA contamination in postharvest fruit fermentation, fourteen coffees that entailed two different postharvest fruit fermentation times were evaluated. These coffees originated in the surroundings of the village of Pedra Menina in the qualified Denomination of Origin and coffee producer region of Caparao on the border between Minas Gerais and Espirito Santo states in Brazil. All coffees were classified according to the Specialty Coffee Association (SCA) protocol and 12 achieved specialty level. OTA was determined in all 14 coffees using immunoaffinity for sample clean-up and high-performance liquid chromatography with fluorescence detection for quantification. One sample presented an OTA concentration of 0.75 µg kg-1 and two samples showed OTA concentrations of 0.87 µg kg-1. The other samples had concentrations of OTA below the limit of quantification obtained in this work (0.64 µg kg-1). Thus, all samples showed OTA concentrations far below the most stringent maximum residue limit (MRL) of 5 µg kg-1 established for roasted coffees by European legislation. These low levels were similar to most of the previous results for Brazilian coffees listed and tabled in this work. This comparison showed that OTA contamination due to this kind of postharvest process - fruit fermentation - should not be a concern for producers and consumers of these fermented coffees.

The aim of this study was to determine the effects of deoxynivalenol (DON) contaminated diet on performance, immune system, gut morphology and jejunal gene expression in broiler chickens. Eighty-one-day old chicks were randomly allotted into two treatments with 4 replicates (10 birds in each replication). Experimental diets were the control diet (maize-soybean meal) and an experimentally contaminated diet with 10 mg/kg DON. The results indicated that DON-challenged birds had decreased (P 0.05) average feed intake (AFI) during starter period as compared to control group. Also, average daily gain (ADG), AFI and feed conversion ratio (FCR) were not affected (P 0.05) by inclusion of DON contaminated diet during the whole experimental period. Dietary addition of DON to the basal diet caused Fabricius bursa relative weight reduction, while increased the abdominal fat and serum triglyceride (TG) concentration (P 0.05). Dietary DON feeding caused an enhancement (P 0.05) in the blood aspartate aminotransferase (AST) and gamma glutamytransferase (GGT) contents. Moreover, DON decreased the serum total protein (TP) and albumin (ALB) concentrations. Inclusion of DON in diet reduced (P 0.05) the white blood cell (WBC) count, lymphocyte number and antibody titer against Newcastle disease virus, but increased (P 0.05) the blood heterophil count. The DON consumption also diminished (P 0.05) the villus height, villus to crypt ratio, mucosa thickness and villus surface area in the duodenum. Mucin-2 expression was decreased (P 0.05) by DON consumption, but toll-like receptor-4 (TLR-4) and claudin-5 (CLDN-5) expressions were not affected (P 0.05) by dietary treatments. In conclusion, although DON could not influence the performance attributes in broiler chickens, it adversely affected the immune response, muc-2 gene expressions in the jejunum and gut morphology, enhanced the liver enzyme indices and lessened the blood protein contents.

Background Aspergillus species cause aflatoxin contamination in groundnut kernels, being a health threat in agricultural products and leading to commodity rejection by domestic and international markets. Presence of Aspergillus flavus and A. parasiticus colonizing groundnut in eastern Ethiopia, as well as presence of aflatoxins have been reported, though in this region, no genetic studies have been done of these species in relation to their aflatoxin production. Results In this study, 145 Aspergillus isolates obtained from groundnut kernels in eastern Ethiopia were genetically fingerprinted using 23 Insertion/Deletion (InDel) markers within the aflatoxin-biosynthesis gene cluster (ABC), identifying 133 ABC genotypes. Eighty-four isolates were analyzed by Ultra-Performance Liquid Chromatography (UPLC) for in vitro aflatoxin production. Analysis of genetic distances based on the approximately 85 kb-ABC by Neighbor Joining (NJ), 3D-Principal Coordinate Analysis (3D-PCoA), and Structure software, clustered the isolates into three main groups as a gradient in their aflatoxin production. Group I, contained 98% A. flavus, including L- and non-producers of sclerotia (NPS), producers of B1 and B2 aflatoxins, and most of them collected from the lowland-dry Babile area. Group II was a genetic admixture population of A. flavus (NPS) and A. flavus S morphotype, both low producers of aflatoxins. Group III was primarily represented by A. parasiticus and A. flavus S morphotype isolates both producers of B1, B2 and G1, G2 aflatoxins, and originated from the regions of Darolabu and Gursum. The highest in vitro producer of aflatoxin B1 was A. flavus NPS N1436 (77.98 μg/mL), and the highest producer of aflatoxin G1 was A. parasiticus N1348 (50.33 μg/mL), these isolates were from Gursum and Darolabu, respectively. Conclusions To the best of our knowledge, this is the first study that combined the use of InDel fingerprinting of the ABC and corresponding aflatoxin production capability to describe the genetic diversity of Aspergillus isolates from groundnut in eastern Ethiopia. Three InDel markers, AFLC04, AFLC08 and AFLC19, accounted for the main assignment of individuals to the three Groups; their loci corresponded to aflC (pksA), hypC, and aflW (moxY) genes, respectively. Despite InDels within the ABC being often associated to loss of aflatoxin production, the vast InDel polymorphism observed in the Aspergillus isolates did not completely impaired their aflatoxin production in vitro.

Alternaria molds are known to cause the contamination of food with their secondary metabolites, a chemically very heterogeneous group of compounds. Yet, after decades of research on the occurrence and the toxicity of Alternaria toxins in academia, no regulation has been implemented yet, thus leaving these potential food contaminants in the status of so-called "emerging mycotoxins". However, research on this topic has been far from static, leading to the European Food Safety Authority repeatedly calling for more data on the occurrence and toxicity of genotoxic metabolites such as alternariol (AOH) and its monomethyl ether (AME). To give an overview on recent developments in the field, this comprehensive review summarizes published data and addresses current challenges arising from the chemical complexity of Alternaria's metabolome, mixture effects and the emergence of novel biological targets like cell membranes or the interaction with different receptors. Besides toxicodynamics, we review recent research on toxicokinetics, including the first in vivo studies which incorporated the rarely investigated-but highly genotoxic-perylene quinones. Furthermore, a particular focus lies on the advances of liquid chromatography/tandem mass spectrometry (LC-MS/MS)-based analytical tools for determining a broader spectrum of Alternaria toxins including modified/masked forms and assessing exposure via human biomonitoring (HBM).

During the period from late 2019 to early 2020, we performed a foodborne virus detection from shellfish collected in Singapore at retail level. Multiple human enteric viruses were included as our targets including human noroviruses (NoVs) GI and GII, hepatitis A virus, hepatitis E virus and rotavirus. Out of the 60 shellfish samples, 23 (38·3%) were detected to be positive by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) with human enteric viruses. Six samples were selected to proceed with virome capture sequencing with positive control samples spiked with serially diluted NoV GII clinical samples in oyster extract. As a result, the natural sample with comparable Ct values (34·0-35·0) of the spiked sample as detected by RT-qPCR generated much lower read counts (>7-log2 cumulative sum scaling difference) and genome coverage (406 nt. vs 3715 nt.), suggesting that the RT-qPCR positive signals detected from the shellfish samples collected at the retail market were likely from degraded RNA derived from inactive virus particles.

Transboundary movement of animal feed and feed ingredients has been identified as a route for pathogen incursions. While imports of animals and animal-derived products are highly regulated for the purpose of infectious disease prevention, there has been less consideration of the viability of infectious agents in inanimate products, such as feed. This study investigated the ability of foot-and-mouth disease virus (FMDV) to remain infectious as a contaminant of commercial whole pig feed and select pig feed ingredients, and to establish the minimum infectious dose (MIDF ) required to cause foot-and-mouth disease (FMD) in pigs that consumed contaminated feed. FMDV viability in vitro varied depending on virus strain, feed product, and storage temperature, with increased duration of infectivity in soybean meal compared to pelleted whole feed. Specifically, both strains of FMDV evaluated remained viable through to the end of the 37 day observation period in experimentally contaminated soybean meal stored at 4 or 20°C . The MIDF for pigs consuming contaminated feed varied across virus strains and exposure duration in the range of 106.2 to 107 TCID50 . The ability of FMDV to cause infection in exposed pigs was mitigated by pre-treatment of feed with two commercially available feed additives, based on either formaldehyde (SalCURB®) or lactic acid (Guardian™). Our findings demonstrate that FMDV may remain infectious in pig feed ingredients for durations compatible with transoceanic transport. Although the observed MIDF was relatively high, variations in feeding conditions and biophysical characteristics of different virus strains may alter the probability of infection. These findings may be used to parameterize modelling of the risk of FMDV incursions and to regulate feed importation to minimize the risk of inadvertent importation.

Background Alternaria toxins are ubiquitous contaminants in highly consumed food products. Therefore, they are candidates to be regulated by EU legislation. In this context, the availability of reliable analytical methods is a keystone both for protecting the health of citizens and smooth functioning of the European market. Objective This paper describes an advanced LC-MS/MS method based on isotope dilution quantification suitable for the determination of altenuene, alternariol, alternariol monomethyl ether, tenuazonic acid, and tentoxin in tomato puree, wheat, and sunflower seeds. Methods The method has been validated in an interlaboratory study that included the analysis of both spiked and naturally contaminated food commodities. Twenty-three participants contributed with analytical data. Results The average recoveries and relative standard deviations for repeatability and reproducibility obtained across the tested matrixes were: 97, 8.0, and 23%, for altenuene, respectively; 95, 9.2, and 17% for alternariol, respectively; 98, 6.4, and 13% for alternariol monomethyl ether, respectively; 97, 4.2, and 9.3% for tenuazonic acid, respectively; and 102, 5.6, and 15% for tentoxin, respectively. The method enabled the determination of all tested Alternaria toxins close to or below 1 µg/kg. Conclusion Overall, the method showed a satisfactory trueness and precision, complying with the requirements for the monitoring of mycotoxins in food in the EU. It is currently under evaluation by the European Committee for Standardization for adoption as a standard method. Highlights Isotope dilution mass spectrometry method for the determination of Alternaria toxins in food.

To evaluate the pollution level of toxic elements in edible mushrooms from Jilin Province, China, the lead, cadmium, mercury, and arsenic content present in them were monitored and evaluated. A total of 610 edible mushroom samples, including fresh and dried, were collected from nine cities in the Jilin Province. The concentrations of lead, cadmium, mercury, and arsenic ranged from 0.007 to 3.31, 0.006 to 48.52, 0.003 to 0.56, and 0.008 to 57.34 mg/kg, respectively, in dried samples, and 0.007 to 0.06, 0.006 to 0.17, 0.003 to 0.06, and 0.008 to 0.12 mg/kg, respectively, in fresh samples. The concentration of cadmium and arsenic exceeded the standard (Cd ≤ 0.2 mg/kg, Cd of Lentinus edodes ≤ 0.5 mg/kg, and As ≤ 0.5 mg/kg) in some mushroom samples. The bioconcentration levels of lead, cadmium, mercury, and arsenic depended on the edible mushroom species, environment, and the physicochemical properties of lead, cadmium, mercury, and arsenic. The results of the single factor pollution index, a method used to calculate the level of single elements in substances, showed that all edible mushrooms were at the level of unpolluted except Tricholoma matsutake and Pleurotus eryngii. The comprehensive factor (P) for T. matsutake was 1.093. The comprehensive factor pollution index, a method used to calculate the level of combined mixture in substances, was in the unpolluted level, except for T. matsutake. The pollution index results showed that cadmium and arsenic concentrations were lightly polluting in some mushroom samples. The health risk index for arsenic was the highest. Therefore, more attention should be paid to arsenic contamination in T. matsutake and P. eryngii in the Jilin Province, China. PRACTICAL APPLICATION: In this paper, we evaluated the contamination levels and associated safety issues of four toxic elements, lead, cadmium, mercury, and arsenic, in edible mushrooms. The results showed that cadmium and arsenic concentrations were lightly polluting in some mushroom samples.

Background Filamentous fungi are the main contamination agent in the viticultural sector. Use of synthetic fungicides is the regular answer to these contaminations. Nevertheless, due to several problems associated with the use of synthetic compounds the industry demands new and safer methods. In the present work, the biopreservation potential of four lactic acid bacteria (LAB) strains was studied against the principal grape contaminant fungi. Results Agar diffusion test evidenced that all four culture-free supernatant (CFS) had antifungal properties against all tested fungi. The Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC) test values evidenced that media fermented by the Lactobacillus plantarum E3 and Lactobacillus plantarum E4 strains showed the highest antifungal activity, resulting in an MFC from 6.3 to 100 g L-1 . Analysis of CFS evidenced the presence of different antifungal compounds, such as lactic acid, phenyllactic acid and pyrazines. In tests on red grapes, an average reduction of 1.32 Log10 of the spores per gram of fruit was achieved by all CFS in grapes inoculated with Aspergillus ochraceous and by 0.94 Log10 for L. plantarum E3 CFS against Botrytis cinerea. Conclusion The antifungal activity of the fermented CFS by L. plantarum E3 reduced the growth of B. cinerea and A. ochraceous in grapes, which are the main contaminant and main productor of ochratoxin A in these crops, respectively. Therefore, based on the results obtained in this work use of the strain L. plantarum E3 can be an interesting option for the biopreservation of grapes. This article is protected by copyright. All rights reserved.

In this study, 241 vegetable-oil food samples were collected from the Hangzhou market in China and analysed for fatty acid esters of 3- and 2-monochloropropanediol (3-MCPD and 2-MCPD) using non-derivative gas chromatography tandem mass spectrometry (GC-MS/MS). Food consumption data were taken from a food consumption survey of urban and rural residents in Hangzhou city performed in 2010-2011. Levels of 3-MCPD esters in edible oil ranged from not detected to 7.98 mg/kg, and the highest mean levels were found in tea seed oil, with concentrations of 2.94 mg/kg. Esters of 2-MCPD levels ranged from not detected to 4.03 mg/kg, and the highest mean levels were also found in tea seed oil, containing 1.49 mg/kg. The range of mean dietary intake of 3-MCPD esters in different groups of edible oil was from 0.096 to 1.54 μg/kg body weight (bw) per day, which is lower than the tolerable daily intake (TDI) established by the European Food Safety Authority (EFSA) (2 μg/kg bw/day). For people aged above 6 years old, the dietary intake of 3-MCPD from edible oil was 0.42 μg/kg bw per day (mean) and 1.22 μg/kg bw per day (P97.5). The range of mean dietary intake of 2-MCPD esters in different groups of edible oil was from 0.025 to 0.79 μg/kg bw/day, and 2-MCPD esters intake was 0.20 μg/kg bw per day (mean) and 0.60 μg/kg bw per day (P97.5). In addition, the dietary intake exposure to 3-MCPD and 2-MCPD esters for urban residents was lower than that for rural residents. The findings indicate that the potential health risks caused by dietary 3-MCPD esters from edible oils were of low concern for most of the Hangzhou residents. However, the exposure risk for consumers with excessive consumption of certain kind of edible oil calls for attention.

Background Honey is a naturally sweet syrup made by honeybees from floral nectar. However, high-fructose corn syrup has been prevalently used for the adulteration of honey. A novel molecular method was developed for the characterization of corn syrup-adulterated honey by specific amplification and quantification of maize residual DNA in honey. An ultra-rapid real-time polymerase chain reaction (UR-qPCR) system for rapid amplification and protocol for direct purification of residual DNA from honey were described. Results Rapidity of maize DNA amplification was acquired within 20 min for a limit of detection of around three copies of targeted DNAs. The amplification of maize residual DNA in honeys adulterated with corn syrup from 5% to 80% (v/v) showed that a minimum rate of 10% adulteration can be identified, and Maize genomic DNA in 5 mL of adulterated honeys was from 13 ± 9 copies to 2478 ± 827 copies, respectively. However, the residual DNA of maize was also detected in natural honey produced in the region where pollen and nectar of maize were collected, and the quantity of maize genomic DNA in these natural honeys was in the range of 10% adulteration with corn syrup. Therefore, detection of both pollen and residual DNA of maize in honey is important in identifying the source of maize residual DNA present in honey. Conclusion A rapid PCR assay was first developed for the accurate detection and quantification of maize residual DNA in honey. It is a useful tool for specific identification of the corn syrup used for honey adulteration. Further studies on residual DNA in various types of corn syrup and specificity of primer are recommended. © 2021 Society of Chemical Industry.

This study aimed to evaluate levels of pesticide residues commonly used on tomato crops in Western Algeria. It showed a significant use of pesticides, especially insecticides (47%) and fungicides (41%) among listed pesticides in crop fields. A total of 30 tomato samples were analysed using multi-residue analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) after QuEChERS extraction. Five pesticides (difenoconazole, triadimenol, metalaxyl, chlorpyriphos and methomyl) were detected in 14 samples (47%). Among these five samples containing residues of two different types of pesticides. The levels ranged from 2.29 to 64.3 µg/kg. The methomyl residue in one sample was above the maximum limit (MRLs) set by the European Union. Our results indicate an excessive use of pesticides and undeniable non-compliance with good agricultural practices. Routine monitoring for pesticides is recommended to preserve consumer's health.

Mercury (Hg) bioaccumulation in fish poses severe threats to the food safety and human health. This study was conducted to assess Hg bioaccumulation in fish (n = 24) and scalp hair (n = 77) of the fishing communities at up- and downstream of the river Swat, Pakistan. The mean Hg concentration in upstream fish Salmo trutta fario (Brown trout) and Schizothorax plagiostomus (Swati fish) species was 34.7±18 μg kg−1 and 29.4±15 μg kg−1, respectively. The mean Hg concentration in downstream Swati fish, Crossocheilus diplochilus (Spena deqa), and Garra gotyla (Tora deqa) was 65±21 μg kg−1, 123±33 μg kg−1, and 326±53 μg Kg−1, respectively. The mean Hg concentration in scalp hair of the up- and downstream fishing communities was 658±125 μg kg−1 and 3969±791 μg kg−1, respectively. Independent T-test showed significant difference (p < 0.001) in the mean Hg concentration in scalp hair of the up- and downstream communities. The most prevalent health problems found in the fishing community were muscle pain, headache, visual impairment, arterial blood pressure, anemia, and kidney dysfunction. Multiple linear regression indicated that daily and weekly consumption of the fish significantly increase Hg accumulation in human scalp hair. Regular consumption of fruits and cruciferous and leafy vegetables were found to reduce Hg toxicity in the population. Further studies are recommended to identify the sources of Hg and welfare impact of fish contamination on the fishing community of river Swat.

Bamboo shoot is an indispensable vegetable in Southeastern China, so a survey of heavy metals in bamboo shoots is a relevant topic of interest. The present study sought to analyse the content of seven heavy metals (As, Cd, Cr, Cu, Mn, Pb and Zn) in six bamboo shoot species from Southeastern China. Percentages of 3.8% (Cd) and 8.9% (Pb) of the samples exceeded the maximum limits as established by the Chinese legislation. Further health risk assessment exhibited that the Cd intake contribution of these samples could not be negligible. The hazard index (HI) at average exposure was less than one. The average HI values in moso bamboo shoot were significantly higher than those of other bamboo shoot species. Overall, the results indicated that the intake of bamboo shoots is safe for consumers in Southeastern China.

A database of 392 pesticides established by an Ultrahigh performance liquid chromatography-tandem Q/Orbitrap high-resolution mass spectrometer (UPLC-Q/Orbitrap-HRMS) was used to screen multiple residues of pesticides in fruit and raw eaten vegetables from planting farms in Shanghai. Risk assessment was conducted with the screened results of the determined pesticides as to evaluate food safety. In 95% of the samples, one or more pesticides had a content below the maximum residue limits (MRLs) as set in the national Chinese standard. The co-occurrence of multi-residues of pesticides was more severe in peach and muskmelon, when compared with other food. All hazard index values of different groups were in the range of 0.19% to 12.3%, demonstrating that chronic dietary risk of studied fruits and raw eaten vegetables is low and the studied food samples were safe for human consumption in terms of these detected pesticides.

The mycotoxin patulin is primarily produced as a secondary metabolite by numerous fungal species and predominantly by Aspergillus, Byssochlamys, and Penicillium species. It is generally associated with fungal infected food materials. Penicillium expansum is considered the only fungal species liable for patulin contamination in pome fruits, especially in apples and apple-based products. This toxin in food poses serious health concerns and economic threat, which has aroused the need to adopt effective detection and mitigation strategies. Understanding its origin sources and biosynthetic mechanism stands essential for efficiently designing a management strategy against this fungal contamination. This review aims to present an updated outline of the sources of patulin occurrence in different foods and their biosynthetic mechanisms. It further provides information regarding the detrimental effects of patulin on human and agriculture as well as its effective detection, management, and control strategies.

The sensitivity of pigs to deoxynivalenol (DON) might be increased by systemic inflammation (SI), which also has consequences for hepatic integrity. Liver lesions and a dys-regulated gene network might hamper hepatic handling and elimination of DON whereby the way of initiation of hepatic inflammation might play an additional role. First and second-pass exposure of the liver with LPS for triggering a SI was achieved by LPS infusion via pre- or post-hepatic venous route, respectively. Each infusion group was pre-conditioned either with a control diet (0.12 mg DON/kg diet) or with a DON-contaminated diet (4.59 mg DON/kg diet) for 4 wk. Liver transcriptome was evaluated at 195 min after starting infusions. DON exposure alone failed to modulate the mRNA expression significantly. However, pre- and post-hepatic LPS challenges prompted transcriptional responses in immune and metabolic levels. The mRNAs for B-cell lymphoma 2-like protein 11 as a key factor in apoptosis and IFN-γ released by T cells were clearly up-regulated in DON-fed group infused with LPS post-hepatically. On the other hand, mRNAs for nucleotide binding oligomerization domain containing 2, IFN-α and eukaryotic translation initiation factor 2α kinase 3 as ribosomal stress sensors were exclusively up-regulated in control pigs with pre-hepatic LPS infusion. These diverse effects were traced back to differences in TLR4 signalling.

Background Thiamethoxam is widely used to control pests in Chinese kale, popularly consumed leafy vegetables. The potential risk to the environment and human health has aroused much public concern. Therefore, it is important to investigate the degradation behavior, residue distribution and dietary risk assessment of thiamethoxam in Chinese kale. Results A sensitive analytical method for determination of thiamethoxam and its metabolite clothianidin residue in Chinese kale was established and validated through a quick, easy, cheap, effective, rugged, and safe (QuEChERS) technique with ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The recoveries were 85.4-101.2% for thiamethoxam and 79.5-108.1% for clothianidin, with the relative standard deviations (RSDs) of 0.9-10.2% and 1.8-6.0%, respectively. For the dissipation kinetics, the data showed that thiamethoxam in Chinese kale was degraded with the half-lives of 4.1 to 4.5 days. In the terminal residue experiments, the residues of thiamethoxam were 0.017-0.357 mg kg-1 after application 2-3 times with a preharvest interval (PHI) of 7 days under the designed dosages. The chronic and acute dietary exposure assessment risk quotient (RQ) values of thiamethoxam in Chinese kale for different Chinese consumers were 0.08-0.19% and 0.05-0.12%, respectively, and those of clothianidin were 0.01-0.04% and 0.02-0.04%, respectively, all of the RQ values were lower than 100%. Conclusion Thiamethoxam in Chinese kale was rapidly degraded following first-order kinetics models. The dietary risk of thiamethoxam and clothianidin through Chinese kale was negligible to consumers. The results from this study are important reference for Chinese governments to developing criteria for the safe and rational use of thiamethoxam, setting maximum residue levels (MRLs), monitoring the quality safety of agricultural products and protecting consumer health. © 2021 Society of Chemical Industry.

The study aimed to develop and evaluate a multiplex polymerase chain reaction assay (mPCR) for the concurrent detection of three major mycotoxin metabolic pathway genes, namely tri8 (T-2 toxin), tri6 (trichothecene) and pks4 (zearalenone), along with competitive internal amplification control. Specific primers for each of the aforementioned genes were optimized and validated using 14 reference strains and 10 pure culture isolates. The optimized mPCR assay detected the three metabolic pathway genes in artificially contaminated maize samples with a sensitivity of 2 × 103 CFU per g for tri6 and pks4 positive Fusarium strains, whereas 2 × 104 CFU per g for tri8 positive Fusarium strains. Application of the developed mPCR assay to 30 cereal and 20 feed samples revealed 24% (12 of 50) contamination with either one or more mycotoxins. The results of mPCR assay were further evaluated with high performance liquid chromatography (HPLC), and both methods provided unequivocal results. This mPCR assay might be a supplementary tool to conventional mycotoxin analytical techniques like thin-layer chromatography, HPLC, etc. The current mPCR assay is a rapid and reliable tool for simultaneous, sensitive and specific detection of T-2, zearalenone and trichothecene producing Fusarium spp. from naturally contaminated foods and to monitor them during the processing steps of food and feed commodities.

Food processors invest significant resources into environmental sampling to detect contamination with potential pathogens, particularly Listeria monocytogenes. To facilitate these efforts, multiple environmental sampling tests (ESTs) have been developed and commercialized that minimize workload, turnaround time, and cost while providing convenient colorimetric detection. For presumptive-positive ESTs, we hypothesized that a relatively minor additional investment could provide, in addition to species confirmation, valuable strain typing data for tracking pathogen spread through a facility, identifying harborage sites, and distinguishing sporadic from persistent or resident contaminants. This hypothesis is based on the demonstrated compatibility of polymorphic locus sequence typing (PLST) with crude samples including food enrichments. Five Listeria ESTs were tested here: broth-based InSite (Hygiena), Path-Chek (Mericon), and Pathfinder (Hardy Diagnositics); and gel-based Petrifilm (3M) and HardyChrom (Hardy Diagnostics). ESTs were inoculated with strains representing two common L. monocytogenes serotypes and nonpathogenic Listeria innocua. Following incubation, broths or suspended colonies were heat treated to inactivate bacteria. Lysates or purified DNAs were prepared and used as templates in PCRs targeting the previously described PLST loci LmiMT1 and LisMT2. Single clear products were obtained from all inoculated ESTs; uninoculated controls were negative. PCR products were subjected to Sanger sequencing, yielding high-quality chromatograms. Phylogenetic analysis confirmed identities to previously determined sequences and revealed relatedness to serotype-matched strains represented in GenBank databases. PRACTICAL APPLICATION: Multiple environmental sampling tests have been commercialized in recent years to facilitate the proactive detection of pathogens, particularly Listeria monocytogenes, within food processing facilities. Coupling a positive detection test with strain typing would enhance its value by providing data that can be used to track pathogen spread through a facility, identify harborage sites, and distinguish sporadic from resident contamination.

Fresh produce imported by Qatar are mostly sold at the wholesale produce market (WPM) located in open-air and near major animal markets and slaughterhouses. This study was the first in Qatar to monitor the effect of environmental conditions on the microbial quality and safety of fresh produce sold at the WPM over 1 year. The monitoring involved the collection of 540 produce samples along with samples of air, soil, and surface swabs. Samples were analyzed for total aerobic bacteria (TAB); generic Listeria spp., Staphylococcus spp., Salmonella spp.; total coliforms and total fungi. Bacterial and fungal isolates were identified using 16S rRNA/ITS rRNA markers. Environmental/sanitary factors significantly impacted the prevalence of microorganisms in all samples tested. Produce quality was rated 'poor' during the months of November-February or May-August, with TAB and coliform counts exceeding 6 and 4 log10 CFU/g, respectively. Bacillus subtilus, Enterobacter cloacae, E. faecium, P. expansium, P. aurantiocandidum, and A. niger were the most abundant species with prevalence rate of 11-30%. The high microbial load of environmental samples indicates that the location of the WPM near livestock markets is likely impacting the microbial quality of fresh produce. Therefore, effective control measures need to be implemented at WPM to improve produce safety yearlong.

We previously examined the associations between dietary dichlorodiphenyldichloroethylene (DDE) and polychlorinated biphenyls (PCBs) intake from fish consumption and type 2 diabetes (T2D) prevalence in Ontario and Manitoba. This study aims to further explore the relationship in a regionally representative sample of First Nations adults living on-reserve across Canada.Dietary, health and lifestyle data collected by the cross-sectional First Nations Food, Nutrition and Environment Study (2008-2018) were analyzed. This participatory study included 6091 First Nations adult participants who answered questions on T2D. The consumption of locally caught fish was estimated with a food frequency questionnaire. A total of 551 samples from 96 fish species were collected and analyzed for the presence of DDE and PCBs. The associations between fish and dietary DDE/PCBs intake with self-reported T2D were investigated using multiple logistic regression models adjusted for confounders.Dietary exposure to DDE (2.11 ng/kg/bw) and PCBs (1.47 ng/kg/bw) vs no exposure was positively associated with T2D with ORs of 2.33 (95% CI: 1.24-4.35) for DDE and 1.43 (95% CI: 1.01-3.59) for PCBs. The associations were stronger among females (DDE OR = 3.11 (1.41-6.88); PCBs OR = 1.76 (1.10-3.65)) and older individuals (DDE OR = 2.64 (1.12-6.20); PCBs OR = 1.44 (1.01-3.91)) as compared with males and younger participants. Also, significant dose-response relationships were found for fish consumption in females only.This study confirms our previous findings that dietary DDE/PCBs exposure may increase the risk of T2D. The effect of DDE/PCBs from fish consumption is driven by geographical differences in DDE/PCBs concentrations in fish and by the amount of fish consumed, and is more prominent in females than in males.RéSUMé: OBJECTIF: Nous avons précédemment examiné les associations entre l’apport alimentaire de dichlorodiphényldichloroéthylène (DDE) et de polychlorobiphényles (PCB) provenant de la consommation de poisson et la prévalence du diabète de type 2 (DT2) en Ontario et au Manitoba. Cette étude vise à explorer davantage la relation dans un échantillon régionalement représentatif d’adultes des Premières Nations vivant dans des réserves partout au Canada. MéTHODE: Les données sur l’alimentation, la santé et le mode de vie recueillies par l’Étude transversale sur l’alimentation, la nutrition et l’environnement chez les Premières Nations (2008–2018) ont été analysées. Cette étude participative comprenait 6 091 participants adultes des Premières Nations qui ont répondu à des questions sur le DT2. La consommation de poisson pêché localement a été estimée à l’aide d’un questionnaire de fréquence alimentaire. Au total, 551 échantillons de 96 espèces de poissons ont été prélevés et analysés pour la présence de DDE et de PCB. Les associations entre la consommation de poisson et l’exposition aux DDE/PCB avec le DT2 auto-déclaré ont été étudiées à l’aide de modèles de régression logistique multiples ajustés pour les facteurs de confusion. RéSULTATS: L’exposition alimentaire au DDE (2,11 ng/kg/pc) et aux PCB (1,47 ng/kg/pc) par rapport à l’absence d’exposition était positivement associée au DT2 avec des OR de 2,33 (IC à 95% : 1,24–4,35) pour le DDE et 1,43 (IC à 95% : 1,01–3,59) pour les PCB. Les associations étaient plus fortes chez les femmes (DDE OR = 3,11 (1,41–6,88); PCB OR = 1,76 (1,10–3,65)) et les individus plus âgés (DDE OR = 2,64 (1,12–6,20); PCB OR = 1,44 (1,01–3,91)) par rapport aux hommes et aux participants plus jeunes. De plus, des relations dose-réponse significatives ont été trouvées pour la consommation de poisson chez les femmes seulement. CONCLUSION: Cette étude confirme nos conclusions précédentes selon lesquelles l’exposition à travers l’alimentation aux DDE/PCB peut augmenter le risque de DT2. L’effet du DDE/PCB sur la consommation de poisson est lié aux différences géographiques dans les concentrations de DDE/PCB dans le poisson et à la quantité de poisson consommée, et est plus important chez les femmes que chez les hommes.

A microcosm study was conducted at two different temperatures under laboratory conditions to investigate the regulatory capacity and the interactive performance of two soil fauna species (Aporrectodea caliginosa, earthworms, and Proisotoma minuta, collembolans) on the reduction of Fusarium toxins in contaminated maize stubbles. Single and mixed species treatments were exposed to artificially infected maize stubbles highly contaminated with the mycotoxins deoxynivalenol (DON) (10,462 µg kg−1) and zearalenone (ZEN) (2,780 µg kg−1) at 17 °C and 25 °C for time periods of 3 and 6 weeks. Immediately after the respective end of incubation, the microcosms were heavily watered to determine the leaching potential of DON and ZEN from contaminated maize stubbles. Maize residues, soil, and eluted water (percolate) samples were analysed for mycotoxin content using liquid chromatography coupled to mass spectrometry. The biomass of introduced earthworms and number of collembolans were monitored to get information about their adaptability to the experimental conditions. While the decline of ZEN was temperature-dependent, but not influenced by faunal activities, a reduction of DON due to faunal impact was observed by trend. In the leaching experiment, 67–82% of the DON content in the residual maize stubbles leached from the plant material by irrigation and was detected in the soil (1.9–3.4 µg kg−1) and in the percolate (12–295 µg L−1). In the case of ZEN, 27–50% of the mycotoxin leached from the residual maize stubbles due to watering but was only occasionally detected in traces in the soil and not found in the percolate. The results clearly reveal a leaching potential of both DON and ZEN, respectively, but a mobilisation with water was only observed for DON. Temperature confirmed to be a key factor, affecting the fate of the mycotoxins in the soil by driving the interaction between different soil fauna members as well as functional and trophic levels within the soil food web.

The objective of this work was to evaluate the microbiological quality of cheese produced by formal and informal micro-enterprises in Paipa, Colombia, to isolate potentially pathogenic bacteria and to determine their prevalence and resistance to antimicrobials such as antibiotics and biocides. Sixteen micro-enterprises of the seventy existing in the region were sampled during 3 years. Viable concentrations of aerobic mesophiles, total and fecal coliforms, Salmonella sp., Listeria monocytogenes, Staphylococcus sp., yeasts, and molds were determined. Seventy-three bacterial isolates were identified by 16S rRNA gene sequencing. The susceptibility of the isolates to antibiotics and biocides was determined. The results indicated that between 98 and 100% of the cheese samples (n = 48 samples) of formal and informal micro-enterprises presented populations of total and fecal coliforms and Staphylococcus sp. above the limits established by Colombian regulations and varied according to the micro-enterprise. The results also indicated that 56% of Staphylococcus isolates were S. aureus. L. monocytogenes was positive in 38% of the samples. Salmonella sp. was not detected. The coliforms that prevailed were Escherichia coli (25%), Citrobacter freundii (14%), and Proteus mirabilis (8%). All L. monocytogenes were sensitive to ampicillin but resistant to erythromycin and trimethoprim-sulfamethoxazole. S. aureus isolates were susceptible to most antibiotics, except tetracycline and erythromycin (7% resistance). Likewise, 30% of coliforms (n = 36) were multidrug-resistant to antibiotics but susceptible to biocides.

In this study, concentrations of 6 different elements (copper (Cu), tin (Sn), zinc (Zn), iron (Fe), cadmium (Cd) and lead (Pb)) in 9 different processed seafood (marinated and smoked mackerel, smoked mackerel, marinated octopus tentacles, marinated octopus salad, marinated squid salad, salted and dried mackerel, marinated and smoked anchovy, smoked salmon and smoked bonito) randomly taken from markets in Mugla province (West Anatolia, Turkey) were determined with the method of inductively coupled plasma mass spectroscopy (ICP-MS) after microwave wet digestion process. In addition, risk assessment for consumer health was conducted by the determination of estimated daily intake (EDI), target hazard quotient (THQ), total target hazard quotient (TTHQ) and carcinogenic risk (CR). According to the obtained results, the seafood product samples with the highest concentrations of each metal were marinated and smoked anchovy for Fe (65.85 ± 16.03 mg kg-1), marinated and smoked anchovy for Zn (64.58 ± 25.16 mg kg-1), marinated octopus salad for Cu (26.33 ± 17.76 mg kg-1), smoked mackerel for Sn (0.42 ± 0.18 mg kg-1), smoked mackerel for Pb (0.28 ± 0.18 mg kg-1) and marinated squid salad for Cd (0.31 ± 0.12 mg kg-1). According to risk assessment for consumer health, it was found that EDI results did not exceed the tolerable daily intake (TDI) values. As a conclusion, any risk for consumer health in terms of CR, THQ and TTHQ values was not detected in the examined samples.