AIEgens enabled ultrasensitive point-of-care test for multiple targets of food safety: Aflatoxin B 1 and cyclopiazonic acid as an example
Food safety is currently a significant issue for human life and health. Various fluorescent nanomaterials have been applied in the point-of-care test (POCT) for food safety as labeling materials. However, previous fluorescent nanomaterials can cause aggregation-caused quenching (ACQ), thus reducing the detection sensitivity. Conversely, aggregation-induced emission luminogens (AIEgens) are promising candidates for POCT in the food safety field because they can enhance detection sensitivity and throughput. Mycotoxins, such as aflatoxin B1 (AFB1) and cyclopiazonic acid (CPA), are a primary threat to human life and health and a significant food safety issue, and their on-site detection from farm to table is needed. Herein, an ultrasensitive point-of-care test was developed based on TPE-Br, a blue-emissive tetraphenylethylene derivative AIEgen. Under optimal conditions, this AIEgen-based lateral-flow biosensor (ALFB) allowed for a rapid response of 8 min toward AFB1 and CPA detection, with considerable sensitivities of 0.003 and 0.01 ng/mL in peanut matrices, respectively. In peanut matrices, the recoveries were 90.3%-110.0% for both mycotoxins, with relative standard deviations (RSDs) below 6%. The ALFB was further validated via UPLC-MS/MS using spiked peanut samples. AIEgens open an avenue for on-site, ultrasensitive, high-throughput detection methods and can be extensively used in point-of-care tests in food safety.
https://doi.org/10.1016/j.bios.2021.113188
Effect of Temperature during Drying and Storage of Dried Figs on Growth, Gene Expression and Aflatoxin Production
Dried fig is susceptible to infection by Aspergillus flavus, the major producer of the carcinogenic mycotoxins. This fruit may be contaminated by the fungus throughout the entire chain production, especially during natural sun-drying, post-harvest, industrial processing, storage, and fruit retailing. Correct management of such critical stages is necessary to prevent mould growth and mycotoxin accumulation, with temperature being one of the main factors associated with these problems. The effect of different temperatures (5, 16, 25, 30, and 37 °C) related to dried-fig processing on growth, one of the regulatory genes of aflatoxin pathway (aflR) and mycotoxin production by A. flavus, was assessed. Firstly, growth and aflatoxin production of 11 A. flavus strains were checked before selecting two strains (M30 and M144) for in-depth studies. Findings showed that there were enormous differences in aflatoxin amounts and related-gene expression between the two selected strains. Based on the results, mild temperatures, and changes in temperature during drying and storage of dried figs should be avoided. Drying should be conducted at temperatures >30 °C and close to 37 °C, while industry processing, storage, and retailing of dried figs are advisable to perform at refrigeration temperatures (<10 °C) to avoid mycotoxin production.
https://doi.org/10.3390/toxins13020134
Spatial analysis of dietary exposure of aflatoxins in peanuts and peanut oil in different areas of China
Peanuts in China are heavily contaminated with aflatoxin, which pose a threaten to human health. To compare the dietary exposure risk of aflatoxins (AFT) in peanuts and peanut oil in different areas of China, the spatial distribution of AFT contamination levels in peanuts and peanut oil from different areas was analyzed. The dietary exposure was calculated by simple distributed risk assessment method before characterizing the health risk using both the margin of exposure (MOE) approach proposed by the European Food Safety Authority (EFSA) and the quantitative liver cancer risk approach proposed by the Joint Food and Agricultural Organization/World Health Organization (FAO/WHO) Expert Committee on Food Additives (JECFA). The results showed that the AFT content in peanuts and peanut oil was high with agglomeration in several provinces of East and South China under a subtropical temperate monsoon climate, and the AFT contamination in peanut oil was more substantial than peanuts. On average, the estimated dietary exposure to AFT from the total of peanuts and peanut oil for Chinese general population ranged from 1.776 to 1.940 ng/kg bw/day (LB-UB), from which the MOE values of 88-96 (UB-LB) and liver cancer risk of 0.055-0.060 cases/100,000 persons/year (LB-UB) were calculated. As for different areas in China, the mean AFT exposure ranged between 0.000 and 17.270 ng/kg bw/day. Moreover, the corresponding health risk was estimated at 10-868759 MOE values and 0.000-0.851 liver cancer cases/100,000 persons/year. Guangdong, Fujian and Jiangxi provinces were at a higher risk rank. The liver cancer risk of AFT exposure from peanuts and peanut oil was far below all-cause liver cancer incidence (18.0 cases/100,000 persons/year) in China, but several areas with relatively high risk should be of concern. Compared with other age groups, children aged 2-6 years should be paid more attention because they have the highest AFT exposure level.
Aflatoxin reduction in nuts by roasting, irradiation and fumigation: a systematic review and meta-analysis
The present study aimed to investigate the reduction of aflatoxins as a potent hazard for human health in nuts during roasting, irradiation, and fumigation processes. A systematic search was performed in PubMed, Scopus, and ISI Web of Science on 6 April 2020 to find interventional studies assessing the effects of roasting, fumigation, and irradiation methods on total and individual aflatoxins concentration in nuts. Study-specific results were pooled by using a random-effects model. A total of 19 trials were included in the analyses. In most studies, the influence of method on aflatoxin reduction was assessed on peanuts. The results showed that the roasting method significantly reduced aflatoxin B1, B2, G1, and G2 concentrations by 46.91%, 30.66%, 40.88%, and 26.19%, respectively. Such results for the fumigation method were 20.88% and 22.56% for aflatoxin B1 and aflatoxin total, respectively. There was a 58.60% reduction in aflatoxin B1 and a 74.97% reduction in aflatoxin total concentrations in nuts following the irradiation method. The findings indicated that the evaluated processes could be influential for reducing aflatoxin levels in nuts.
Aflatoxins in foodstuffs: occurrence and risk assessment in Turkey
This study aims to determine dietary exposure to aflatoxin B1 (AFB1) and the sum of aflatoxins (AFT) for the Turkish population and assess the risks to human health related to aflatoxins. A total of 1473 results on five main food categories commercialised in Turkey in 2002–2019 were used in the risk assessment. In the adult population, the mean middle bound (MB) exposure levels were 0.433 ng kg−1 b.w. per day for AFB1 and 0.511 ng kg−1 b.w. per day for AFT. The 95th percentile dietary exposure of AFB1 and AFT was estimated at 1.19 and 1.29 ng kg−1 b.w. per day, respectively. Pistachios (44.4 %) made the highest contribution to mean AFB1 exposure in Turkish adults, followed by maize/maize flour (16.2 %), groundnuts (13.8 %), chilli (10.1 %), walnuts (4.9 %), hazelnuts (4.0 %), chocolate (3.9 %) and others (2.7 %). The margin of exposure (MOE) estimates for mean and 95th percentile exposures to AFB1 are considerably lower than 10 000, which raises a potential health concern for Turkish adults.
Analysis of E.U. Rapid Alert System (RASFF) Notifications for Aflatoxins in Exported U.S. Food and Feed Products for 2010-2019
The most common, toxic, and carcinogenic mycotoxins found in human food and animal feed are the aflatoxins (AFs). The United States is a leading exporter of various nuts, with a marketing value of $9.1 billion in 2019; the European Union countries are the major importers of U.S. nuts. In the past few years, border rejections and notifications for U.S. tree nuts and peanuts exported to the E.U. countries have increased due to AF contamination. In this work, we analyzed notifications from the "Rapid Alert System for Food and Feed (RASFF)" on U.S. food and feed products contaminated with mycotoxins, primarily AFs, for the 10-year period 2010-2019. Almost 95% of U.S. mycotoxin RASFF notifications were reported for foods and only 5% for feeds. We found that 98.9% of the U.S. food notifications on mycotoxins were due to the AF contamination in almond, peanut, and pistachio nuts. Over half of these notifications (57.9%) were due to total AF levels greater than the FDA action level in food of 20 ng g-1. The Netherlands issued 27% of the AF notifications for U.S. nuts. Border rejection was reported for more than 78% of AF notifications in U.S. nuts. All U.S. feed notifications on mycotoxins occurred due to the AF contamination. Our research contributes to better understanding the main reasons behind RASFF mycotoxins notifications of U.S. food and feed products destined to E.U. countries. Furthermore, we speculate possible causes of this problem and provide a potential solution that could minimize the number of notifications for U.S. agricultural export market.
Computational modelling of survival of Aspergillus flavus in peanut kernels during hot air-assisted radio frequency pasteurization.
In recent years, radio frequency (RF) heating is getting popular as an alternative pasteurization method for agricultural commodities and low moisture foods. Computer simulation is an effective way to help understand RF interactions with food components and predict temperature distributions among food samples after RF treatments. In this study, a computer model based on Joule heating and thermal inactivation kinetic of A. flavus was established to predict both temperature distribution and microbial reduction among peanut kernels after RF processing. For the process validation, three 2-g peanut samples inoculated with 40 μL A. flavus were placed at three representative locations among 2.17 kg peanut kernels and subjected to various processing conditions in a 27.12 MHz, 6 kW RF heating unit together with hot air system. Results showed that the average difference of the sample temperature and microbial reduction between simulation and experiment was small with RMSE values of 0.009 °C and 0.012 °C, and 0.31 log CFU/g and 0.42 log CFU/g for peanut moisture contents of 7.56% and 12.02% w. b., respectively. Nonuniform RF heating resulted in the least lethality of A. flavus at the cold spot. The validated computer model was further used to estimate microbial reduction distributions at other target temperatures based on predicted temperature profiles. This computer model may help design the RF pasteurization protocols for peanut kernels without extensive experiments in food industry.
The Occurrence of Aflatoxins in Nuts and Dry Nuts Packed in Four Different Plastic Packaging from the Romanian Market.
Mycotoxins are secondary metabolites produced by various fungi. A very important category of mycotoxins are aflatoxins, considered to be the most dangerous in humans. Aflatoxin B1, well known as a favorable factor in the occurrence of hepatocellular carcinoma in humans, is the most controversial of all mycotoxins. Aflatoxins, found in naturally contaminated food, are resistant to degradation by heat. Current food processing practices and conventional storage conditions do not completely eliminate aflatoxin contamination from the food supply chain. Long storage food products-such as peanuts, pistachio, nuts in general, and dried fruits-are susceptible to aflatoxins contamination. The type of plastic material can influence the concentration of aflatoxins during storage due to the permeability to gas and moisture exchange with the external milieu. Nuts in general and dried fruits are consumed in large quantities worldwide. Therefore, herein we investigated the effect of plastic material on the total aflatoxins and aflatoxin B1 content in 64 samples of nuts and dried fruits packed and stored in low-density polyethylene (LDPE), polypropylene (PP), polyethylene (PE), and polyethylene terephthalate (PET). The method consisted in a cleanup procedure using immunoaffinity columns coupled with RIDASCREEN FAST immunoenzymatic competitive assays based on the ELISA technique. Collected data were subjected to statistical analysis and multiple comparisons tests were applied. From the total analyzed samples, 14.06% exceeded the maximum admitted European levels for total aflatoxins. The highest concentrations of total aflatoxins were obtained from samples packed in LDPE, followed by PP, PE, and PET. Aflatoxin B1 was detected in all samples packed in LDPE, PP, and PE. Most of the samples packed in PET had concentrations <1 µg/kg. These results indicate that nuts in general packed and stored in LDPE are more prone to contamination with aflatoxins, while PET is more suitable for maintaining the quality and safety of these products.
Development of PCR, LAMP and qPCR Assays for the Detection of Aflatoxigenic Strains of Aspergillus flavus and A. parasiticus in Hazelnut.
Aspergillus flavus and A. parasiticus are two species able to produce aflatoxins in foodstuffs, and in particular in hazelnuts, at harvest and during postharvest phase. As not all the strains of these species are aflatoxin producers, it is necessary to develop techniques that can detect aflatoxigenic from not aflatoxigenic strains. Two assays, a LAMP (loop-mediated isothermal amplification) and a real time PCR with TaqMan® probe were designed and validated in terms of specificity, sensitivity, reproducibility, and repeatability. The capability of the strains to produce aflatoxins was measured in vitro and both assays showed to be specific for the aflatoxigenic strains of A. flavus and A. parasiticus. The limit of detection of the LAMP assay was 100-999 picograms of DNA, while the qPCR detected 160 femtograms of DNA in hazelnuts. Both techniques were validated using artificially inoculated hazelnuts and naturally infected hazelnuts. The qPCR was able to detect as few as eight cells of aflatoxigenic Aspergillus in naturally infected hazelnut. The combination of the LAMP assay, which can be performed in less than an hour, as screening method, with the high sensitivity of the qPCR, as confirmation assay, is able to detect aflatoxigenic strains already in field, helping to preserve the food safety of hazelnuts.
Removal of aflatoxin B1 from contaminated peanut oils using magnetic attapulgite
The efficient magnetic adsorbent (Fe3O4@ATP) was prepared by precipitation through the dispersion of Fe3O4 nanoparticles on the natural attapulgite (ATP) and then tested as an adsorbent for aflatoxin B1 (AFB1) removal from contaminated oils. The adsorbent characterization results revealed that the Fe3O4 were incorporated into the ATP, affording the Fe3O4@ATP composite. This magnetic composite displayed a good ability to eliminate AFB1 from contaminated oils with a removal efficiency of 86.82% using a 0.3% dosage. The Fe3O4@ATP possessed paramagnetic character with a saturation magnetization of 50.86 emu/g, enabling its easy separation from the medium using an external magnet. The adsorption process followed the pseudo-second-order model and fitted the Freundlich isotherm well. Moreover, the thermodynamic studies showed that AFB1 adsorption onto Fe3O4@ATP was exothermic and spontaneous. The novelty of this study lies in the fabrication of magnetic composite adsorbents for AFB1 elimination from oils.