Unraveling the fate of mycotoxins during the production of legume protein and other derived products.

Consumers are increasingly looking for healthier and sustainable diets. Plant-based diets rich in legumes satisfy this demand. Legumes contain protein, dietary fibers and starch. Technological processes can separate these fractions, which can be used as supplements, or as ingredients. Nonetheless, legumes are susceptible to fungal infection, causing a potential health concern, since some fungi can produce mycotoxins: toxic secondary metabolites. The aim of this work was to analyze the fate of mycotoxins during different stages of the production process of legume derived products from the raw materials to final products. An extraction followed by liquid chromatography-tandem mass spectrometry was used for the analysis, revealing the presence of enniatin B (ENN B), alternariol monomethyl ether (AME), deoxynivalenol, T2-toxin, nivalenol, fumonisin B1 and sterigmatocystin in raw materials, intermediate products and side streams. The alkaline solubilization steps, were effective in reducing ENN B; however, AME was found in one of the final products.

Results of multi-actor collaboration in risk analysis: a simplified risk assessment toolkit for rapid detection of emerging risks.

The dynamic field of food safety faces continuous challenges, prompting stakeholders to develop collaborative actions for improved food safety systems. As part of these actions, the EU-FORA fellowship programme was dedicated to a multi-actor collaboration addressing risks of the unregulated mycotoxins T-2 and HT-2 toxins in oats. Critical gaps in risk assessment procedures were identified, leading to a joint effort to develop a strategy for rapid data collection and risk assessment, including the development of a risk assessment toolkit comprising of a training manual and two intuitive Microsoft® Excel files. The toolkit enables efficient data collection and processing, facilitating risk assessment calculations and rapid risk detection. Applying the toolkit to assess T-2 and HT-2 toxin risks in Belgian oats revealed minimal concerns, except for children aged 3-9 years, likely due to an overestimation. The toolkit is available on the FoodSafety4EU Platform and will be refined based on user feedback, promoting better risk assessment practices. This approach empowers stakeholders, from professionals to policymakers, fostering collaboration and enhancing food safety practices.

European food safety research: An explorative study with funding experts' consultation.

The European research area exhibits considerable opacity and fragmentation in food safety research funding and organizational structures, impeding the exploitation of existing research potential across European countries. Given that food safety is inherently linked to the societal challenges of our time, identifying and removing existing barriers to research funding in this area is crucial. Towards investigating this matter, interviews were conducted with funding bodies from six European countries to assess key issues related to research funding in general and food safety in particular. Funding experts were then invited to a workshop to jointly discuss the challenges identified and explore strategies to address them. Evaluation of the food safety research funding situation in selected European countries revealed both convergences and significant differences among national funding bodies. Engaging with funding experts provided invaluable insights into the issues encountered with research funding, such as inadequate call management staff or insufficient research funds, culminating in a set of recommendations for action to remedy the situation.

Dietary exposure assessment and risk characterization of citrinin and ochratoxin A in Belgium.

Exposure to mycotoxins is a worldwide problem. To ensure public health, it is imperative to characterize the risks related to these toxins. The present study aims to conduct a dietary exposure assessment of citrinin (CIT) and ochratoxin A (OTA) in the Belgian population using consumption data of a variety of foodstuffs. A total of 367 food samples from different food categories were collected in Belgian supermarkets and analysed for CIT and OTA using a validated liquid chromatography-tandem mass spectrometry method. Daily CIT and OTA exposure to the Belgian population was calculated based on the analytical results and food consumption data in three age categories (3-9, 10-17 and 18-64 years), obtained from a national food consumption survey. Furthermore, a risk characterization was performed for CIT, in which no intake values exceeded the tolerable daily intake (TDI) of 200 ng kg-1 bw day-1, indicating no health risk. However, a CIT intake level of 187 ng kg-1 bw day-1 was detected for children in the age category of 3-9 years in the worst case scenario for rice, indicating that rice consumption could contain a potential health hazard for young children. For OTA, a potential health risk was detected in several food categories (biscuits, croissants, rice, flour, meat imitates, herbs and spices) in the higher percentiles (P99) or at maximum found concentrations when calculating the margin of exposure (MoE) for neoplastic effects. An attempt to perform a cumulative health risk assessment for both toxins was done. Although a high number of uncertainties is involved, combined margin of exposure (MoET) values indicated a potential health risk related to the combined exposure to CIT and OTA. For the first time, our study demonstrated the potential health risks of CIT and OTA after individual and combined exposure, in particular related to rice consumption. Moreover, further research is recommended concerning multiple mycotoxin exposure in young children.

Instant killing of pathogenic chytrid fungi by disposable nitrile gloves prevents disease transmission between amphibians.

To prevent transmission of the pathogenic chytrid fungi Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal), hygiene protocols prescribe the single use of disposable gloves for handling amphibians. We discovered that rinse water from nitrile gloves instantly kills 99% of Bd and Bsal zoospores. Transmission experiments using midwife toads (Alytes obstetricans) and Bd, and Alpine newts (Ichthyosaura alpestris) and Bsal, show that the use of the same pair of gloves for 2 subsequent individuals does not result in significant transmission of any chytrid fungus. In contrast, handling infected amphibians bare-handed caused transmission of Bsal in 4 out of 10 replicates, but did not result in transmission of Bd. Based on the manufacturer's information, high resolution mass spectrometry (HRMS) and colorimetric tests, calcium lactate and calcium nitrate were identified as compounds with antifungal activity against both Bd and Bsal. These findings corroborate the importance of wearing gloves as an important sanitary measure in amphibian disease prevention. If the highly recommended single use of gloves is not possible, handling multiple post-metamorphic amphibians with the same pair of nitrile gloves should still be preferred above bare-handed manipulation.

Comprehensive toxicokinetic analysis reveals major interspecies differences in absorption, distribution and elimination of citrinin in pigs and broiler chickens.

A comprehensive toxicokinetic analysis of citrinin (CIT) revealed interspecies differences for all toxicokinetic parameters and in absolute oral bioavailability. Oral bioavailability for CIT was complete for broilers (113-131%), while ranging from 37 to 44% in pigs. CIT was more rapidly absorbed in pigs (Tmax = 0.92 h) compared to broiler chickens (Tmax = 7.33 h). The elimination of CIT was slower in pigs (T1/2el = 26.81 h after intravenous (IV) administration) compared to chickens (T1/2el = 1.97 h after IV administration), due to the striking difference in clearance (Cliv=9.87 mL/h/kg for pigs versus Cliv = 863.09 mL/h/kg for broilers). Also, the volume of distribution differed significantly between pigs (Vd = 0.30 L/kg after IV administration) and chickens (Vd = 2.46 L/kg after IV administration). However, plasma protein binding did not differ statistically significant (91-98%). It is imperative to further investigate biotransformation and elimination pathways in different species, including humans.

A Study of Carry-Over and Histopathological Effects after Chronic Dietary Intake of Citrinin in Pigs, Broiler Chickens and Laying Hens.

Citrinin (CIT) is a polyketide mycotoxin occurring in a variety of food and feedstuff, among which cereal grains are the most important contaminated source. Pigs and poultry are important livestock animals frequently exposed to mycotoxins, including CIT. Concerns are rising related to the toxic, and especially the potential nephrotoxic, properties of CIT. The purpose of this study was to clarify the histopathological effects on kidneys, liver, jejunum and duodenum of pigs, broiler chickens and laying hens receiving CIT contaminated feed. During 3 weeks, pigs (n = 16) were exposed to feed containing 1 mg CIT/kg feed or to control feed (n = 4), while 2 groups of broiler chickens and laying hens (n = 8 per group) received 0.1 mg CIT/kg feed (lower dose group) and 3 or 3.5 mg CIT/kg feed (higher dose group), respectively, or control feed (n = 4). CIT concentrations were quantified in plasma, kidneys, liver, muscle and eggs using a validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method. Kidneys, liver, duodenum and jejunum were evaluated histologically using light microscopy, while the kidneys were further examined using transmission electron microscopy (TEM). Histopathology did not reveal major abnormalities at the given contamination levels. However, a significant increase of swollen and degenerated mitochondria in renal cortical cells from all test groups were observed (p < 0.05). These observations could be related to oxidative stress, which is the major mechanism of CIT toxicity. Residues of CIT were detected in all collected tissues, except for muscle and egg white from layers in the lowest dose group, and egg white from layers in the highest dose group. CIT concentrations in plasma ranged between 0.1 (laying hens in lower dose group) and 20.8 ng/mL (pigs). In tissues, CIT concentrations ranged from 0.6 (muscle) to 20.3 µg/kg (liver) in pigs, while concentrations in chickens ranged from 0.1 (muscle) to 70.2 µg/kg (liver). Carry-over ratios from feed to edible tissues were between 0.1 and 2% in pigs, and between 0.1 and 6.9% in chickens, suggesting a low contribution of pig and poultry tissue-derived products towards the total dietary CIT intake for humans.

Development and validation of an LC-MS/MS method for the simultaneous determination of citrinin and ochratoxin a in a variety of feed and foodstuffs.

An ultra-performance liquid chromatography-electrospray tandem mass spectrometry (UPLC-ESI+/--MS/MS) method for the simultaneous analysis of citrinin (CIT) and ochratoxin A (OTA) in feed (chicken and pig) and food (cereal-based products, fruit, vegetable juices, nuts, seeds, herbs, spices, vegetarian and soy products, alcoholic beverages, baby food products and food supplements) was developed. The mycotoxins were extracted from these matrices using a QuEChERS-based extraction method without any further clean-up step. The samples were 5-fold concentrated. Final extracts were analyzed using a UPLC-MS/MS system and chromatographic separation was achieved by applying a gradient elution for a total run time of 10 min. Mycotoxins were quantified using an internal calibration via analyte/13C-labeled internal standard ratio. The developed method was validated according to the criteria described in Commission Regulation No. 401/2006/EC and Commission Decision No. 2002/657/EC. Specificity, linearity, apparent recovery, limit of detection and quantification, intraday and interday precision, measurement uncertainty, matrix effect, and extraction efficiency were the parameters studied. Finally, 90 Belgian chicken and pig feed samples were analyzed, revealing the simultaneous presence of CIT (