A 10-year analysis of RASFF notifications for mycotoxins in nuts. Trend in key mycotoxins and impacted countries.

The demand for tree nuts has significantly grown in recent years as epidemiological studies and clinical intervention trials demonstrated an inverse relationship between tree nut consumption and chronic diseases. However, mycotoxins are one of the main hazards responsible for increased "Rapid Alert System for Food and Feed" (RASFF) notifications and border rejections on nuts and nut products exported to the E.U. countries in the past few years. Mycotoxins are secondary metabolites that present serious threats to human and animal health. The most prevalent, toxic, and carcinogenic mycotoxins observed in human food and animal feed are the aflatoxins (AFs). This work analyzed notifications from the RASFF on nuts and nut products contaminated with mycotoxins, for a 10-year period from 2011 to 2021. A total of 4752 mycotoxin notifications were published on RASFF for food products worldwide, 63% (n = 3000) were notified in "nuts, nut products and seeds". It was observed that 95% (n = 2669) notifications were due to AFs. Over half of these notifications (52%, n = 1545) were reported for groundnuts, where 29% (n = 441) of the notifications were received for groundnuts from China alone. Border rejection was reported for 91% (n = 2560) of the nuts and nut products which received the notifications from the E.U. countries. This study proffers understanding into the major reasons for RASFF notifications on nuts and nut products exported to E.U. countries. Also, the implications of this issue with some recommendations that could reduce the incidents of notifications for tree nuts have been outlined.

A model framework to communicate the risks associated with aflatoxins.

Risk communication is defined as the interactive exchange of information and opinions concerning risk, risk-related factors and risk perceptions amongst all the stakeholders of food safety throughout the risk analysis process. The interactive exchange of information occurs at three different levels i.e. informed level, dialogue level and engagement level. For an effective food safety risk communication (FSRC), it is important that the information should adhere to the core principles of risk communication which are transparency, openness, responsiveness and timeliness. Communication of a food safety risk within all the components of risk communication strategy constitutes a complex network of information flow that can be better understood with the help of a framework. Therefore, a model framework to communicate the risks associated with aflatoxins (AFs) dietary intake has been developed with the aim of (a) creating general awareness amongst public and (b) involving industry stakeholders in the prevention and control of risk. The framework has been motivated by the learnings and best practices outlined in the identified technical guidance documents for risk communication. Risk assessors, risk managers, industry stakeholders and general public have been identified as the major stakeholders for the present framework. Amongst them, industry stakeholders and general public has been selected as the major target audience for risk managers. Moreover, population residing in low- and middle-income countries (LMIC) has been identified as the main target group to reach.

The importance and mitigation of mycotoxins and plant toxins in Southeast Asian fermented foods.

Fermented foods (ffs) and beverages are widely consumed in Southeast Asia (SEA) for their nutritional balance, flavor, and food security. They serve as vehicles for beneficial microorganisms performing a significant role in human health. However, there are still major challenges concerning the safety of ffs and beverages due to the presence of natural toxins. In this review, the common toxins found in traditional ffs in SEA are discussed with special reference to mycotoxins and plant toxins. Also, mitigation measures for preventing risks associated with their consumption are outlined. Ochratoxin, citrinin, aflatoxins were reported to be major mycotoxins present in SEA ffs. In addition, soybean-based ff food products were more vulnerable to mycotoxin contaminations. Common plant toxins recorded in ffs include cyanogenic glycosides, oxalates, phytates and saponins. Combined management strategies such as pre-harvest, harvest and post-harvest control and decontamination, through the integration of different control methods such as the use of clean seeds, biological control methods, fermentation, appropriate packaging systems, and controlled processing conditions are needed for the safe consumption of indigenous ffs in SEA.

The Occurrence and Co-Occurrence of Regulated, Emerging, and Masked Mycotoxins in Rice Bran and Maize from Southeast Asia.

Raw feed materials are often contaminated with mycotoxins, and co-occurrence of mycotoxins occurs frequently. A total of 250 samples i.e., rice bran and maize from Cambodia, Laos, Myanmar, and Thailand were analysed using state-of-the-art liquid chromatography-mass spectrometry (LC-MS/MS) for monitoring the occurrence of regulated, emerging, and masked mycotoxins. Seven regulated mycotoxins - aflatoxins, ochratoxin A, fumonisin B1, deoxynivalenol, zearalenone, HT-2, and T-2 toxin were detected as well as some emerging mycotoxins, such as beauvericin, enniatin type B, stachybotrylactam, sterigmatocystin, and masked mycotoxins, specifically zearalenone-14-glucoside, and zearalenone-16-glucoside. Aspergillus and Fusarium mycotoxins were the most prevalent compounds identified, especially aflatoxins and fumonisin B1 in 100% and 95% of samples, respectively. Of the emerging toxins, beauvericin and enniatin type B showed high occurrences, with more than 90% of rice bran and maize contaminated, whereas zearalenone-14-glucoside and zearalenone-16-glucoside were found in rice bran in the range of 56-60%. Regulated mycotoxins (DON and ZEN) were the most frequent mycotoxin combination with emerging mycotoxins (BEA and ENN type B) in rice bran and maize. This study indicates that mycotoxin occurrence and co-occurrence are common in raw feed materials, and it is critical to monitor mycotoxin levels in ASEAN's feedstuffs so that mitigation strategies can be developed and implemented.

The evolution of multiplex detection of mycotoxins using immunoassay platform technologies.

Mycotoxins present serious threats not only for public health, but also for the economy and environment. The problems become more complex and serious due to co-contamination of multiple hazardous mycotoxins in commodities and environment. To mitigate against this issue, accurate, affordable, and rapid multiplex detection methods are required. This review presents an overview of emerging rapid immuno-based multiplex methods capable of detecting mycotoxins present in agricultural products and feed ingredients published within the past five years. The scientific principles, advantages, disadvantages, and assay performance of these rapid multiplex immunoassays, including lateral flow, fluorescence polarization, chemiluminescence, surface plasmon resonance, surface enhanced Raman scattering, electrochemical sensor, and nanoarray are discussed. From the recent literature landscape, it is predicted that the future trend of the detection methods for multiple mycotoxins will rely on the advance of various sensor technologies, a variety of enhancing and reporting signals based on nanomaterials, rapid and effective sample preparation, and capacity for quantitative analysis.

Metataxonomic analysis of bacterial communities and mycotoxin reduction during processing of three millet varieties into ogi, a fermented cereal beverage.

Ogi is a fermented cereal beverage, made primarily from maize (Zea mays) and rarely from millets. Unlike maize-based ogi, little is known about the bacterial community and mycotoxin profile during the production of millet-based ogi. Therefore, the bacterial community dynamics and mycotoxin reduction during ogi processing from three millet varieties were investigated using next-generation sequencing of the 16S rRNA gene and liquid chromatography-tandem mass spectrometry, respectively. A total of 1163 amplicon sequence variants (ASVs) were obtained, with ASV diversity across time intervals influenced by processing stage and millet variety. ASV distribution among samples suggested that the souring stage was more influenced by millet variety than the steeping stage, and that souring may be crucial for the quality attributes of the ogi. Furthermore, bacterial community structure during steeping and souring was significantly differentiated (PERMANOVA, P < 0.05) between varieties, with close associations observed for closely-related millet varieties. Taxonomically, Firmicutes, followed by Actinobacteria, Bacteroidetes, Cyanobacteria and Proteobacteria phyla were relatively abundant (>1%). Lactic acid bacteria, such as Burkholderia-Caballeronia-Paraburkholderia, Lactobacillus, Lactococcus and Pediococcus, dominated most fermentation stages, suggesting their roles as key fermentative and functional bacteria in relation to mycotoxin reduction. About 52-100%, 58-100% and 100% reductions in mycotoxin (aflatoxins, beauvericin, citrinin, moniliformin, sterigmatocystin and zearalenone) concentrations were recorded after processing of white fonio, brown fonio and finger millet, respectively, into ogi. This study provides new knowledge of the dominant bacterial genera vital for the improvement of millet-based ogi through starter culture development and as well, elucidates the role of processing in reducing mycotoxins in millet ogi.

Co-occurrence and toxicological relevance of secondary metabolites in dairy cow feed from Thailand.

The occurrence of secondary metabolites and co-contaminants in dairy cow feed samples (n = 115), concentrate, roughage, and mixed feed, collected from Ratchaburi and Kanjanaburi provinces, Thailand, between August 2018 and March 2019 were investigated using LC-MS/MS based multi-toxin method. A total of 113 metabolites were found in the samples. Fungal metabolites were the predominant compounds, followed by plant metabolites. Among major mycotoxins, zearalenone and fumonisins were most frequently detected in concentrate and mixed feed samples, while deoxynivalenol and aflatoxin B1 were found at the frequency lower than 50%. Other metabolites, produced by Aspergillus, Fusarium, Penicillium, and Alternaria species, occurred in the samples. Flavoglaucin, 3-nitropropionic acid, averufin, and sterigmatocystin were the most prevalent Aspergillus metabolites. Common Fusarium metabolites occurring in the samples included moniliformin, beauvericin, and enniatins. For Penicillium metabolites, mycophenolic acid, questiomycin A, quinolactacin A, oxaline, citrinin, and dihydrocitrinone were frequently detected. The toxic Alternaria metabolites, alternariol, and alternariol monomethyl ether showed the high incidence in the samples. Plant metabolites were commonly found, mainly cyanogenic compounds and isoflavones, from cassava and soybean meal used as feed ingredients. Overall, 96.6% of feed samples contained at least two metabolites, in a range from 2 to 69. According to co-contamination of mycotoxins found in feed samples, zearalenone were mostly found in combination with fumonisin B1, deoxynivalenol, and aflatoxin B1. Fumonisin B1 co-occurred with aflatoxin B1 and deoxynivalenol. The mixtures of deoxynivalenol and aflatoxin B1, and of zearalenone, fumonisin B1 and deoxynivalenol were also found. Due to known individual toxicity of fungal and plant metabolites and possible additive or synergistic toxic effects of multi-mycotoxins, the occurrence of these metabolites and co-contaminants should be monitored continuously to ensure food safety through the dairy supply chain.

In Vitro Mechanism Assessment of Zearalenone Removal by Plant-Derived Lactobacillus plantarum BCC 47723.

Zearalenone (ZEA) is a harmful secondary fungal metabolite, produced primarily by plant pathogenic fungi mostly belonging to the genus Fusarium. It is involved in reproductive disorders in animals since its structure is similar to the estrogen hormone. This induces precocious pubertal changes, fertility problems, and hyper estrogenic disorders. The main objectives of this study were to evaluate the ZEA removal capacity of plant-derived lactic acid bacteria (LAB) and to investigate the possible components and mechanisms involved in the removal of ZEA by physically and chemically treated plant-derived LAB. The bacterial cells were characterized using scanning electron microscopy coupled with energy dispersive X-ray spectroscopy (SEM-EDS), Fourier transform infrared spectroscopy (FTIR), and the analysis of zeta potential, and hydrophobic index. Results revealed that 17 out of 33 plant-derived LAB exhibited ZEA removal from liquid medium. The percentage of removal ranged from 0.5-23% and Lactobacillus plantarum BCC 47723, isolated from wild spider flower pickle (Pag-sian-dorng), exhibited the highest removal. The alteration of proteins on L. plantarum BCC 47723 structure by Sodium dodecyl sulphate (SDS) treatment was positively affected on ZEA removal, whereas that of lipids on ZEA removal was negatively observed. Heat treatment influenced the higher ZEA adsorption. SEM images showed that the morphologies of modified bacterial cells were distinctly deformed and damaged when compared with untreated control. FTIR analysis indicated that the original functional groups, which included amide (C=O, C-N), carboxyl (C=O, C-O, O-H), methylene (C=C), and alcohol (O-H) groups, were not changed after ZEA adsorption. The zeta potential indicated that electrostatic interaction was not involved in the ZEA removal, while hydrophobicity was the main force to interact with ZEA. These findings can conclude that adsorption by hydrophobicity is the main mechanism for ZEA removal of plant-derived L. plantarum BCC 47723. The alteration of bacterial cell structure by heat treatment enhanced the efficiency of L. plantarum BCC 47723 for ZEA reduction. Its activity can be protected by the freeze-drying technique. Hence, plant-derived L. plantarum BCC 47723 can be considered as an organic adsorbent for ZEA reduction in food and feedstuff.

Aflatoxin B1-degrading activity from Bacillus subtilis BCC 42005 isolated from fermented cereal products.

Aflatoxin B1 is a naturally occurring mycotoxin that is produced as secondary metabolite by Aspergillus spp., especially A. flavus and A. parasiticus. This is the most severe toxin due to its carcinogenic, mutagenic, and teratogenic properties. Hence, methods for toxin degradation have been received increasing interest from both scientific communities and industries. In this study, 32 isolates of Bacillus spp. from various fermented cereal products were screened for their aflatoxin B1 degradation ability. The results indicated the extracellular fraction of Bacillus subtilis BCC 42005 isolated from Iru (African locust bean) potentially possessed aflatoxin B1-degrading ability. The maximum activity of the active fraction was at 50°C and pH 8.0. The activity was stable in a wide range of pH (5.0-8.0) and temperature (25-60°C). The aflatoxin B1-degrading mechanisms of this strain may be possibly involved by enzyme(s). This extracellular fraction was not toxic at IC50 4 mg/ml and it can be combined with water as a soaking agent for maize, which results in 54% of aflatoxin B1 reduction after contact time 120 min. Hence, the extracellular fraction of Bacillus subtilis BCC 42005 can be further applied as an effective soaking agent in a pretreatment process with a practical and easy-to-implement condition and also probably used to reduce the aflatoxin B1 contamination in other foods and feeds commodities.

The Effectiveness of Durian Peel as a Multi-Mycotoxin Adsorbent.

Durian peel (DP) is an agricultural waste that is widely used in dyes and for organic and inorganic pollutant adsorption. In this study, durian peel was acid-treated to enhance its mycotoxin adsorption efficacy. The acid-treated durian peel (ATDP) was assessed for simultaneous adsorption of aflatoxin B1 (AFB1), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), and fumonisin B1 (FB1). The structure of the ATDP was also characterized by SEM-EDS, FT-IR, a zetasizer, and a surface-area analyzer. The results indicated that ATDP exhibited the highest mycotoxin adsorption towards AFB1 (98.4%), ZEA (98.4%), and OTA (97.3%), followed by FB1 (86.1%) and DON (2.0%). The pH significantly affected OTA and FB1 adsorption, whereas AFB1 and ZEA adsorption was not affected. Toxin adsorption by ATDP was dose-dependent and increased exponentially as the ATDP dosage increased. The maximum adsorption capacity (Qmax), determined at pH 3 and pH 7, was 40.7 and 41.6 mmol kg-1 for AFB1, 15.4 and 17.3 mmol kg-1 for ZEA, 46.6 and 0.6 mmol kg-1 for OTA, and 28.9 and 0.1 mmol kg-1 for FB1, respectively. Interestingly, ATDP reduced the bioaccessibility of these mycotoxins after gastrointestinal digestion using an in vitro, validated, static model. The ATDP showed a more porous structure, with a larger surface area and a surface charge modification. These structural changes following acid treatment may explain the higher efficacy of ATDP in adsorbing mycotoxins. Hence, ATDP can be considered as a promising waste material for mycotoxin biosorption.

Utilising an LC-MS/MS-based multi-biomarker approach to assess mycotoxin exposure in the Bangkok metropolitan area and surrounding provinces.

Human exposures to mycotoxins through dietary intake are a major health hazard and may result in various pathophysiological effects. Although Thailand is a country at increased risk due to its climatic conditions, no comprehensive dataset is available to perform proper exposure assessment of its population with regard to mycotoxins. Therefore, this pilot study was conducted to investigate and evaluate the exposure levels of major mycotoxins (aflatoxin B1, ochratoxin A, fumonisins, zearalenone and trichothecenes). Sixty first-morning urine samples were collected from healthy volunteers who live in the Bangkok metropolitan area and surrounding provinces (Pathumthani, Nonthaburi, Samutprakarn and Samutsakorn). Urine samples were analysed by a LC-MS/MS-based multi-biomarker method following a so-called 'dilute and shoot' approach. Results generally indicated low mycotoxin exposures in most individuals through the determination of the four biomarkers that were detected in urine samples, i.e. aflatoxin M1, ochratoxin A (OTA), as well as the deoxynivalenol (DON) metabolites DON-3-glucuronide and DON-15-glucuronide in 10 of 60 individuals. The maximum concentrations were used to estimate the daily intake confirming that none of the individuals exceeded the tolerable daily intake (TDI) of DON (maximum 26% of TDI) or OTA (maximum 22% of TDI). However, the maximum exposure of aflatoxin B1, estimated to be 0.91 µg (kg bw)⁻¹ day⁻¹, should raise some concerns and suggests further studies utilising a more sensitive method. Low exposure to Fusarium toxins was also confirmed by the absence of zearalenone, α-zearalanol, ß-zearalanol and zearalenone-14-glucuronide as well as T-2 toxin, HT-2 toxin, nivalenol and free DON. This is the first multi-mycotoxin biomarker study performed in Southeast Asia.