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Aim: This study aims for the first time in Portugal to assess the early-life exposure to mycotoxins through a mother & child cohort. N/A

Deoxynivalenol (DON), T-2 toxin (T-2) and HT-2 toxin (HT-2), due to their incidence and toxicity, are the trichothecenes of major concern for cereals and cereal-based products. Moreover, natural occurrence of modified forms of DON, such as 3-acetyl-deoxynivalenol (3Ac-DON), 15-acetyl-deoxynivalenol (15Ac-DON), and deoxynivalenol-3-glucoside (DON3G), and of T-2 and HT-2, such as T-2 glucoside (T-2G) and HT-2 glucoside (HT-2G), have been reported by several authors. The development of analytical methods for the simultaneous detection of these mycotoxins and their modified forms, also expressed as the sum, is highly requested since it could meet future requirements of European or international regulations. For this reason, two fluorescence polarization immunoassays (FPIAs) have been developed and in-house validated for the rapid (

Human biomonitoring (HBM) is recognized as an important tool to assess the Human exposure to chemicals, contributing to describe trends and patterns of exposure over time and to identify population groups that could be under risk. Natural chemicals as mycotoxins, fungi metabolites that produce toxic effects in humans, are important compounds that could be found in foods usually consumed worldwide in a daily basis. Mycotoxins as aflatoxins (AFTs) are genotoxic, carcinogenic and immunosuppressive compounds. Hepatocellular carcinoma (HCC) is one of their main health toxic effects and is the third leading cause of cancer deaths worldwide. In Portugal, scarce data are available regarding exposure to AFTs and none previous study used HBM data to characterize comprehensively the burden associated to this exposure. In the scope of the National Food, Nutrition, and Physical Activity Survey of the Portuguese General Population (2015-2016), 24h-urine samples from 94 participants were analyzed by liquid chromatography–mass spectrometry (LC-MS/MS) for the simultaneous determination of AFTs (B1, B2, G1, G2, M1). A model was developed to estimate the health impact of the exposure of Portuguese population to aflatoxins, estimating the number of cases of HCC and DALYs attributed to AFTs exposure. AFTs were detected in 12.8% (AFB1), 16.0% (AFB2) and 19.1% (AFM1) of the 24h-urine samples. The estimated number of extra cases of HCC attributed to this exposure ranged from 17 to 65 cases/year; the associated DALYs for the Portuguese population ranged from 284 to 1802 years. The present study generated, for the first time and within a HBM study, reliable data on internal exposure to AFTs at individual level for the Portuguese population. These data were crucial to characterize the burden derived from this exposure and to support risk managers to establish preventive policy measures that contribute to ensure the public health protection. This research was performed with the financial support to CESAM (UID/AMB/50017/2019), to FCT/MEC through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020. This research was also supported by the project MYTOX-SOUTH, Ghent University Global Minds programme. IAN-AF national survey was funded by the EEA Grants Program, Public Health Initiatives (PT06 - 000088SI3) N/A

Alternariol (AOH) and zearalenone (ZEN) are mycotoxins that occur widely in food commodities, with particular incidence in cereals [1,2]. Both mycotoxins present estrogenic effects, therefore, it is crucial to assess the human exposure and the associated risk [2,3]. To date, there is a lack of human studies to assess exposure to ZEN and AOH through biomarkers analysis in Portugal. In the scope of the National Food, Nutrition, and Physical Activity Survey of the Portuguese General Population (IAN-AF, 2015-2016), 94 participants collected 24h-urine samples [4]. Following a salt-assisted matrix extraction, urine samples were analyzed by liquid chromatography–mass spectrometry (LC-MS/MS) for the simultaneous determination of ZEN, α-zearalenol (α-ZEL), β -zearalenol (β-ZEL), α-zearalanol (α-ZAL), β-zearalanol (β-ZAL), zearalanone (ZAN), zearalenone-14-glucoside (ZEN-14-GlcA), α-zearalenol-glucoside (α-ZEL-GlcA), β-zearalenol-glucoside (β-ZEL-GlcA), zearalenone-14-sulfate (ZEN-14-Sulf), alternariol monomethyl ether (AME) and AOH. Risk characterization was performed using the Hazard Quotient (HQ) approach. ZEN, ZEN-14-GlcA and AOH were detected in 48%, 16% and 29% of analyzed samples, respectively, and median levels were 0.17 µg/L for ZEN and ZEN-14-GlcA and 0.28 µg/L for AOH. Participants were exposed to ZEN (n=35), AOH (n=15) and some volunteers showed co-exposure to both mycotoxins (n=12). Risk characterization of exposure to ZEN was performed and 24% of participants presented a HQ > 1, thus exceeding the Tolerable Daily Intake (TDI) of 0.250 µg/kg bw/day. There is no toxicological data available to support the establishment of a TDI for AOH, thus the risk associated with this toxin was not characterized [2,5]. The present study generated, for the first time and within a human biomonitoring study, reliable data regarding the exposure of the Portuguese population to ZEN and AOH. These data are crucial to perform a more realistic risk assessment, contributing to more effective preventive measures for health protection of the Portuguese population. Thanks are due for the financial support to CESAM (UID/AMB/50017/2019), to FCT/MEC through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020. The authors also acknowledge the support of MYTOX-SOUTH project. N/A

Soybean is a critical food and nutritional security crop in Rwanda. Promoted by the Rwandan National Agricultural Research System for both adults and as an infant weaning food, soybean is grown by approximately 40% of households. Soybean may be susceptible to the growth of mycotoxin-producing moulds; however, data has been contradictory. Mycotoxin contamination is a food and feed safety issue for grains and other field crops. This study aimed to determine the extent of mycotoxin contamination in soybean, and to assess people’s awareness on mycotoxins. A farm-level survey was conducted in 2015 within three agro-ecological zones of Rwanda suitable for soybean production. Soybean samples were collected from farmers (n=300) who also completed questionnaires about pre-and post-harvest farm practices, and aflatoxin awareness. The concentration of total aflatoxin in individual soybean samples was tested by enzymelinked immunosorbent assay (ELISA) using a commercially-available kit. Other mycotoxins were analyzed using liquid chromatography-mass spectrometry (LCMS/ MS) on 10 selected sub samples. Only 7.3% of the respondents were aware of aflatoxin contamination in foods, but farmers observed good postharvest practices including harvesting the crop when the pods were dry. Using enzyme-linked immunosorbent assay (ELISA), only one sample had a concentration (11 μg/kg) above the most stringent EU maximum permitted limit of 4 μg/kg. Multi-mycotoxins liquid chromatography-mass spectrometry (LC-MS/MS) results confirmed that soybeans had low or undetectable contamination; only one sample contained 13μg/kg of sterigmatocystine. The soybean samples from Rwanda obtained acceptably low mycotoxin levels. Taken together with other studies that showed that soybean is less contaminated by mycotoxins, these results demonstrate that soybean can be promoted as a nutritious and safe food. However, there is a general need for educating farmers on mycotoxin contamination in food and feed to ensure better standards are adhered to safeguard the health of the consumers regarding these fungal secondary metabolites.Key words: soybean, safety, mould, aflatoxin, mycotoxins, sterigmatocystine, ELISA, LC-MS/MS, Rwanda

According to the General Food Law (EC) No. 178/2002, which sets the legal basis of the European Union food controls, the food and feed business operators have the primary responsibility for the safety of their products. Their demand for simplified and rapid test methods at critical control points in the entire chain has never been greater. Validated rapid screening methods are nowadays recognised as a strategic tool to tackle this issue, and, by analogy with chromatographic methods, mainly based on liquid chromatography-mass spectrometry, the new trend in this field is multiplexing. This topic also falls within the scopes of the MycoKey project, which explored different technologies for rapid multi-mycotoxin screening, including: multiplex strip test immunoassays for the semi-quantitative determination of Fusarium toxins and fluorescence polarization immunoassays for the quantitative simultaneous determination of trichothecenes and their modified forms. The process of evaluating methods fitness-for-purposes according to EC official guidelines and performance criteria (Regulation 519/2014/EU) will be presented and discussed. Validation according to harmonised protocols is critical to establish credibility of results across both scientific and regulatory boundaries. However, besides methods' performances, practicality of the different approaches for mycotoxin screening needs to be considered. These aspects will be discussed in comparison with liquid chromatography-mass spectrometry based methods when applied for mycotoxin screening purposes. Moreover, a MycoKey survey was performed among scientists and stakeholders for the collection and sharing of current knowledge and experience on rapid methods for mycotoxin screening. This, together with some lessons learned during the MycoKey round table discussion on mycotoxin detection methods, enabled a better understanding of trends in research and development areas in this field. The results from these discussions and their significance will be summarised to highlight priorities and critical issues to be addressed to provide multi-mycotoxin rapid methods that fit stakeholders' needs and expectations.

The trichothecenes of major concern for cereals and cereal-based products, due to their incidence and toxicity, are deoxynivalenol (DON), T-2 toxin (T-2) and HT-2 toxin (HT-2). These toxins can be harmful to both human and animal health. Moreover, a large number of modified forms of DON, T-2 and HT-2 generated by fungi, plants and mammals have been isolated and characterized. To date, natural occurrence of modified forms of DON, such as 3-acetyl-deoxynivalenol (3Ac-DON), 15-acetyl-deoxynivalenol (15Ac-DON), and deoxynivalenol-3-glucoside (DON3G), and of T-2 and HT-2, such as T-2 glucoside (T-2G) and HT-2 glucoside (HT-2G), have been reported by several authors. The development of analytical methods able to simultaneously detect mycotoxins and their modified forms, also expressed as the sum, has high impact because it could meet possible future requirements of European or international regulations. For this reason, within the MycoKey Project (EU, H2020), two FPIAs have been developed and validated for the rapid (

International audience; 4,15-Diacetoxyscirpenol (DAS) is a mycotoxin primarily produced by Fusarium fungi and occurring predominantly in cereal grains. As requested by the European Commission, the EFSA Panel on Contaminants in the Food Chain (CONTAM) assessed the risk of DAS to human and animal health related to its presence in food and feed. Very limited information was available on toxicity and on toxicokinetics in experimental and farm animals. Due to the limitations in the available data set, human acute and chronic health-based guidance values (HBGV) were established based on data obtained in clinical trials of DAS as an anticancer agent (anguidine) after intravenous administration to cancer patients. The CONTAM Panel considered these data as informative for the hazard characterisation of DAS after oral exposure. The main adverse effects after acute and repeated exposure were emesis, with a no-observed-adverse-effect level (NOAEL) of 32 lg DAS/kg body weight (bw), and haematotoxicity, with a NOAEL of 65 lg DAS/kg bw, respectively. An acute reference dose (ARfD) of 3.2 lg DAS/kg bw and a tolerable daily intake (TDI) of 0.65 lg DAS/kg bw were established. Based on over 15,000 occurrence data, the highest acute and chronic dietary exposures were estimated to be 0.8 and 0.49 lg DAS/kg bw per day, respectively, and were not of health concern for humans. The limited information for poultry, pigs and dogs indicated a low risk for these animals at the estimated DAS exposure levels under current feeding practices, with the possible exception of fattening chicken. Assuming similar or lower sensitivity than for poultry, the risk was considered overall low for other farm and companion animal species for which no toxicity data were available. In consideration of the similarities of several trichothecenes and the likelihood of co-exposure via food and feed, it could be appropriate to perform a cumulative risk assessment for this group of substances.