Molecularly imprinted polymer based on covalent organic framework coated steel substrate as the mass spectrometric ionization source for the direct detect of aflatoxins in complex food matrices.

Ambient mass spectrometry allows direct analysis of various sample types with minimal or no pretreatment. However, due to the influence of matrix effects, there are sensitivity and issues in analyzing trace analytes in complex food samples. In this work, we developed a spray mass spectrometry platform based on SSS@TPBD-TPA@MIPs (Stainless steel substrate (SSS), terephthalaldehyde (TPA), N, N, N', N'-tetrakis(p-aminophenyl)-p-phenylenediamine (TPBD), molecularly imprinted polymer (MIP)), for rapid, in situ, high-throughput, highly enrichment efficiency and highly selective trace analysis of aflatoxins. By simplifying the sample pretreatment and directly applying high voltage for ESI-MS, the analysis can be completed within 1 min. The established method base on SSS@TPBD-TPA@MIPs exhibited high sensitivity and accuracy when determine trace level AFs in maize and peanuts. The results demonstrated a good linear relationship within the range of 0.01-10 µg/L, with the determination coefficient (R2) ≥ 0.9956. The limits of detection (LODs) was 0.035-0.3 ng/mL and limits of quantitation (LOQs) was 0.12-0.99 ng/mL, with acceptable recovery rate of 82.09-115.66 % and good repeatability represented by the relative standard deviation (RSD) less than 17.43 %. Furthermore, SSS@TPBD-TPA@MIPs exhibited excellent reusability, with more than 8 repeated uses, and showed good adsorption performance.

Simultaneous detection of multiple mycotoxins in Radix Dipsaci and estimation of exposure risk for consumers.

Like many traditional Chinese herbal medicines, preparations from Radix Dipsaci are at risk of contamination by harmful mycotoxins; however, there have been no reports of actual contamination. In this study, we developed an analytical method to simultaneously detect eight mycotoxins in Radix Dipsaci and estimate the exposure risk for consumers. We have developed an analytical method utilizing ultra-high performance liquid chromatography and tandem mass spectrometry to accurately determine the levels of AFB1, AFB2, AFG1, AFG2, OTA, ZEN, T-2 and ST mycotoxins in 45 batches of Radix Dipsaci sourced from major medicinal herb markets across five regions in China. We also analyzed migration of mycotoxins from the raw herbs into water decoction. Based on these results and data on human consumption of the herbal medicine, we estimated risk of exposure and acceptable exposure limits to mycotoxins in the Radix Dipsaci using the "margin of exposure (MOE)" method. Of the 45 batches of Radix Dipsaci, 48.89% contained at least one of the eight mycotoxins, 24.44% contained one, 17.78% contained two and 6.67% contained three. The most frequent mycotoxins were aflatoxin B1, present in 35.56% of batches (at 0.25-34.84 µg/kg); aflatoxin G1, 15.56% (1.99-44.05 µg/kg); and ochratoxin A, 22.22% (16.11-143.38 µg/kg). These three mycotoxins transferred from the raw herb into water decoction at respective rates of 20.20%, 29.14%, and 24.80%. The 95th percentile values of the MOE risk factors for health effects of AFB1 were below 10,000 at high doses but above 10,000 at low doses of Radix Dipsaci long-term treatment. With the reduction in duration of exposure years, the MOE values of AFB1 and AFG1 gradually reverted to within the acceptable range. The mean, 50th, and 95th percentile values of the MOE risk factors for health effects of OTA exceeded 10,000 regardless of whether consumers received a low or high dose of Radix Dipsaci treatment for durations ranging from 1 to lifetime. Based on this exposure and a typical human diet, we have estimated the respective 20-year exposure limits for Radix Dipsaci to be 5.821 µg/kg, 4.035 µg/kg, and 56.073 µg/kg for the three mycotoxins under consideration. Contamination with multiple mycotoxins is frequently observed in Radix Dipsaci, and the three most prevalent contaminants have been found to leach into water decoctions, thereby posing a potential health hazard for individuals consuming this herbal preparation. This work highlights the need to monitor herbal medicines for mycotoxin contamination in order to protect consumers.

Aflatoxins in dried chilli products in Gansu Province and health risk.

In order to understand the status of aflatoxin contamination in dried chilli products in Gansu Province and the risk of dietary exposure, a total of 106 samples of dried chilli products from farmers' markets and supermarkets in 14 prefecture-cities of Gansu Province were collected and analysed by isotope dilution liquid chromatography-tandem mass spectrometry. The results showed that the detection rate of aflatoxin in dried chilli products in Gansu Province was 30.2%, and the average level was 1.57 µg/kg. The detection rates of dried chillies, paprika, and chilli powders were 16.7%, 43.6%, and 46.2%, respectively. The detection rates of aflatoxin in dried chilli products from shops and farmers' markets were 22.5% and 40.0%, respectively. The dietary exposure of AFB1 was 0.0001 µg/kg bw/day, and the MOE calculated from its average concentration was 305.

Mycobiota, Total Aflatoxins, And Ochratoxin A of Black And Green Cumin.

Because of the medical importance of cumin as well as it being one of the food additives to many Saudi dishes, there was a need to study the fungal load of this type of spice. This study aimed to determine the mycological profile of the retail black and green cumin distributed in different markets at western region, Saudi Arabia, using the dilution plat method on dichloran 18% glycerol (DG18) agar and incubation at 25°C. Using morphological criteria and molecular markers (internal transcribed spacer sequence), 39 species belonging to 18 genera were collected from different black cumin (33 species belonging to 17 genera) and green cumin (25 species belonging to 9 genera). Alternaria alternata, Aspergillus flavus, A. niger, A. ochraceus, Cladosporium cladosporioides, and Stemphylium botryosum were the most prevalent. Black cumin harbors fungal counts reaching 545 colony-forming units (CFU)/g, while green cumin included 500 CFU/g. Also, the natural occurrence of aflatoxins and ochratoxin A was also measured. Seventy-two cumin samples (90% of tested samples) showed toxin contamination. Aflatoxins and ochratoxin A ranged from 9.35 to 3.9 PPB in black cumin samples and from 4.08 to 5.75 PPB in green cumin samples.

Climatic effects on aflatoxin contamination of maize.

Aflatoxins are frequent contaminants of maize especially in the face of climate change with deleterious health and socio-economic impacts. South Africa is ranked 9th maize exporter globally; hence, insights need to be gained in terms of the maize value chain in South Africa with respect to aflatoxin contamination to evaluate consumers' exposure. High-performance liquid chromatography (HPLC) technique was used in this study to quantify aflatoxins in South African commercial maize. One thousand and twenty-eight (1028) maize samples were collected across six distinct agro-climatic regions over five harvest seasons (2017 - 2021). A total of 205 samples (19.94 %) were found to be contaminated with aflatoxins, with mean total aflatoxin concentration of 64.17 ppb amongst the contaminated samples, which is above the SA regulatory limit of 20 ppb for animal consumption. The year 2018 recorded the highest mean total aflatoxin value while North-West agro-climatic region had the highest mean total aflatoxin value. Drastic reduction in average rainfall significantly influence aflatoxin contamination of South African maize.

Safe detoxification on acid-washed activated carbon combined with chitosan for aflatoxins from contaminated peanut oil.

Aflatoxins are one of the most toxic mycotoxins and can cause serious harm to humans and animals. Adsorption is a practical decontamination technique favored by the industry because of its advantages of low cost, speed and simplicity, and environmental friendliness. In this work, the adsorption features of activated carbon and chitosan were fabricated in a composite through chemical co-precipitation to improve its properties for adsorption. Furthermore, the capacity of the synthesized chitosan and acid-washed activated carbon composite (CS-AAC) to attenuate the aflatoxins in contaminated peanut oil and the adsorption capacity at different initial aflatoxins content, contact duration, and temperature were evaluated. The results showed a higher adsorption capacity (removal efficiency to 93.45% of AFB1, 94.05% of AFB2, 89.16% of AFG1, 83.26% of AFG2). The Freundlich isothermal and D-R model and the pseudo-second-order rate expression both implied a good correlation with the test data and explained the adsorption mechanism well. The adsorption mechanism was found to be accomplished primarily via ion exchange and chelation. According to thermodynamic results (△G < 0, △H > 0, △S > 0), the adsorption process was endothermic and spontaneous. Compared to acid-washed activated carbon, CS-AAC enhanced the retention of VE and sterols (especially VE by 23%), and the safety of CS-AAC adsorbent was explored by cellular experiments. In conclusion, CS-AAC is a promising adsorbent material for the removal of aflatoxins from edible oils.

Development and validation of a DLLME-HPLC-FLD method for determination of aflatoxins in Chrysanthemum morifolium based on quality by design principles.

INTRODUCTION: Aflatoxins, potent carcinogens produced by Aspergillus species, present significant health risks and commonly contaminate herbal products such as Chrysanthemum morifolium. Detecting these toxins in C. morifolium proves challenging due to the complex nature of the herbal matrix and the fluctuating levels of toxins found in different samples. OBJECTIVES: This study aimed to develop and optimize a novel method for the detection of aflatoxins in C. morifolium using dispersive liquid-liquid microextraction combined with high-performance liquid chromatography-fluorescence detection based on quality by design principles. METHODOLOGY: The method involved determining critical method attributes and parameters through the Plackett-Burman design, followed by optimization using the Box-Behnken design. Monte Carlo simulation was employed to establish a design space, which was experimentally verified. Method validation was performed to confirm accuracy, precision, and stability. RESULTS: The developed method exhibited excellent linearity (R2 > 0.9991) for aflatoxins B1, B2, G1, and G2 across a range of concentrations, with recovery rates between 85.52% and 102.01%. The validated method effectively quantified aflatoxins in C. morifolium under different storage conditions, highlighting the impact of temperature and storage time on aflatoxin production. CONCLUSION: This study successfully established a reliable and effective method for the detection of aflatoxins in C. morifolium, highlighting the importance of strict storage conditions to reduce aflatoxin contamination. Using a quality by design framework, the method demonstrated robustness and high analytical performance, making it suitable for routine quality control of herbal products.

A novel triple-signal biosensor based on ZrFe-MOF@PtNPs for ultrasensitive aflatoxins detection.

The development of more sensitive, stable, and portable biosensors is crucial for meeting the growing demands of diverse and complex detection environments. MOF-based nanozymes have emerged as excellent optical reporters, making them ideal signal donors for constructing multi-signal lateral flow immunoassays (LFIA). In this study, a ZrFe-MOF@PtNPs nanocomposite was synthesized by uniformly depositing platinum nanoparticles (PtNPs) onto the surface of ZrFe-MOFs using an impregnation-reduction method. The ZrFe-MOF@PtNPs exhibited broad absorption spectra, excellent peroxidase-like activity (SA = 21.77 U/mg), high solvent stability, and efficient antibody binding capability. A portable LFIA platform was developed based on ZrFe-MOF@PtNPs and a smartphone for the targeted detection of carcinogenic aflatoxins. This method enabled the readout of colorimetric, fluorescent, and catalytic signals, significantly enhancing detection sensitivity, ensuring result accuracy, and expanding the dynamic detection range. For aflatoxin M1, the calculation of the detection limit of the three signal modes reached as low as 0.0062 ng/mL, which is two orders of magnitude more sensitive than AuNPs-LFIA (0.1839 ng/mL). This study provides effective guidance for multifunctional modification of MOFs and serves as a reference for the application of MOF-based nanozymes in point-of-care biosensors.

Preparation and application of a pseudo-templated multi-monomer aflatoxins imprinted polymer.

A functional material was developed with specific recognition properties for aflatoxins for pre-processing enrichment and separation in the detection of aflatoxins in Chinese herbal medicines. In the experiment, ethyl coumarin-3-carboxylate, which has a highly similar structure to the oxonaphthalene o-ketone of aflatoxin, was selected as a pseudo-template, zinc acrylate, neutral red derivative, and methacrylic acid, which have complementary functions, were selected as co-monomers to prepare a pseudo-template multifunctional monomer molecularly imprinted polymer (MIP). The MIP obtained under the optimal preparation conditions has a maximum adsorption capacity of 0.036 mg/mg and an imprinting factor of 3.67. The physical property evaluation of the polymers by Fourier infrared spectrometer, scanning electron microscopy, pore size analyzer, thermogravimetric analyzer, and diffuse reflectance spectroscopy showed that the MIP were successfully prepared and porous spherical-like particles were obtained. The synthesized polymer was used as a solid-phase extraction agent for the separation of aflatoxins from the extract of spina date seed. The linear range of the developed method was 10-1000 ng/mL, the limit of detection was 0.36 ng/mL, the limit of quantification was 1.19 ng/mL, and the recoveries of the extracts at the concentration level of 0.2 µg/mL were in the range 88.0-93.4%, with relative standard deviations (RSDs) of 1.97% (n). The results showed that the preparation of MIPs using ethyl coumarin-3-carboxylate as a template was simple, economical, and convenient. It is expected to become a promising functional material for the enrichment and separation aflatoxins from complex matrices.

Optimization and Validation of a Cheaper, Safer, and More Sustainable Methodology for Aflatoxins Determination in Rich-Lipidic Matrices (Pistachio Nuts) Using Deep Eutectic Solvent Extraction and UHPLC-FLD Analysis.

Aflatoxins pose a major health concern and require strict monitoring in food products. Existing methods rely on hazardous organic solvents for extraction, prompting the development of a greener alternative. This study explores deep eutectic solvents (DESs) for aflatoxin extraction from pistachios, a valuable food product prone to aflatoxin contamination. The proposed method utilizes DES extraction followed by solid-phase extraction cleanup and ultrahigh-performance liquid chromatography coupled with fluorescence detector analysis. Recovery rates ranged from 85.5 to 99.1% for pistachios spiked with 1-8 ng/g aflatoxins, in compliance with EU regulations, with coefficients of variation less than 2.94%. The method demonstrates good sensitivity with limits of detection and quantification in the range of 0.02-0.22 ng/g and 0.05-0.72 ng/g, respectively. Greenness assessment using AGREEPrep and White Analytical Chemistry metrics confirms its environmental sustainability. This approach offers a promising, safer, and more eco-friendly alternative for aflatoxin extraction from complex food matrices like pistachios.

Rapid Detection of Aflatoxins in Ground Maize Using Near Infrared Spectroscopy.

Aflatoxins are carcinogenic mycotoxins that may contaminate many crops and more especially maize. To protect consumers from these contaminants, many countries set up low regulatory thresholds of few µg/kg. The control of food requires time-consuming analysis for which sampling is a key step. It would therefore of key sanitary and economic relevance to develop rapid, sensitive and accurate methods that could even be applied on line at harvest, to identify batches to be excluded as soon as possible. In this study, we analyzed more than 500 maize samples taken at harvest during 3 years for their aflatoxin contamination using HPLC-MS. Among them, only 7% were contaminated but sometimes at levels largely exceeding European regulations. We demonstrate that Near InfraRed Spectroscopy (NIRS) could be of great help to classify cereal samples according to their level of aflatoxin contamination (below or higher than E.U. regulation). To build the model, all AF contaminated samples as well as an equivalent number of AF free samples were used. NIRS performance was not sufficient to quantify the toxins with adequate precision. However, its ability to discriminate naturally contaminated maize samples according to their level of contamination with aflatoxins in relation to European regulations using a quadratic PCA-DA model was excellent. Accuracy of the model was 97.4% for aflatoxin B1 and 100% for total aflatoxins.

Effective extraction and detection of aflatoxins in cereals using nitrogen-rich benzodiimidazole linkage magnetic covalent organic framework based solid phase extraction and HPLC-MS/MS analysis.

Cereals are frequently contaminated by aflatoxins (AFs). The objective of this study was to develop an efficient extraction materials for rapidly extracting and detecting AFs. A novel amino-functionalized benzodiimidazole linkage magnetic covalent organic framework (Fe3O4@BB-COF) was simply fabricated by one-step cyclization and aromatization. The Fe3O4@BB-COF, having multiple N-containing active sites, exhibited excellent extraction capability towards AFs due to synergistic interactions, including the π-π interactions, hydrogen bonding interactions, polar interactions, electrostatic interactions and Lewis acid-base interactions. The Fe3O4@BB-COF based MSPE method for detecting aflatoxins has advantages of simple operation, short extraction time (6 min), and low material consumption (2 mg). This method exhibited satisfactory linearity (0.05-20 µg/kg), and sensitivity (0.01-0.45 µg/L for the detection limits) and accuracy (76.8-97.1 % for recovery) and was successfully applied for extracting and detecting AFs in cereals.

Pre- and post-harvest aflatoxin contamination and management strategies of Aspergillus spoilage in East African Community maize: review of etiology and climatic susceptibility.

Globally, maize (Zea mays L.) is deemed an important cereal that serves as a staple food and feed for humans and animals, respectively. Across the East African Community, maize is the staple food responsible for providing over one-third of calories in diets. Ideally, stored maize functions as man-made grain ecosystems, with nutritive quality changes influenced predominantly by chemical, biological, and physical factors. Food spoilage and fungal contamination are convergent reasons that contribute to the exacerbation of mycotoxins prevalence, particularly when storage conditions have deteriorated. In Kenya, aflatoxins are known to be endemic with the 2004 acute aflatoxicosis outbreak being described as one of the most ravaging epidemics in the history of human mycotoxin poisoning. In Tanzania, the worst aflatoxin outbreak occurred in 2016 with case fatalities reaching 50%. Similar cases of aflatoxicoses have also been reported in Uganda, scenarios that depict the severity of mycotoxin contamination across this region. Rwanda, Burundi, and South Sudan seemingly have minimal occurrences and fatalities of aflatoxicoses and aflatoxin contamination. Low diet diversity tends to aggravate human exposure to aflatoxins since maize, as a dietetic staple, is highly aflatoxin-prone. In light of this, it becomes imperative to formulate and develop workable control frameworks that can be embraced in minimizing aflatoxin contamination throughout the food chain. This review evaluates the scope and magnitude of aflatoxin contamination in post-harvest maize and climate susceptibility within an East African Community context. The paper also treats the potential green control strategies against Aspergillus spoilage including biocontrol-prophylactic handling for better and durable maize production.

Nutritional conditions affecting of selenium nanoparticles synthesized by Fusarium oxysporum (CCASU-2023-F9), and their biological activities against mycotoxin-producing fungi isolated from animal feed.

One of the most promising biologically based nanomanufacturing processes is the production of selenium nanoparticles (SeNPs) by fungi. The use of these biosynthesized nanoparticles in agricultural practices has emerged as a new approach for controlling pathogen growth and mycotoxin production. In the present study, different chemical and physical parameters were investigated for the growth of Fusarium oxysporum (CCASU-2023-F9) to increase selenite reduction and obtain the highest yield of selenium nanoparticles (SeNPs). Fusarium oxysporum (CCASU-2023-F9) exhibited tolerance to up to 1 mM sodium selenite (Na2SeO3), accompanied by red coloration of the medium, which suggested the reduction of selenite and the formation of selenium nanoparticles (SeNPs). Reduced selenite was quantified using inductively coupled plasma‒mass spectrometry (ICP-MS), and the results revealed that Fusarium oxysporum (CCASU-2023-F9) is able to transform 45.5% and 50.9% of selenite into elemental selenium by using fructose and urea as the best carbon and nitrogen sources, respectively. An incubation temperature of 30 °C was the best physical condition at which 67.4% of the selenite was transformed into elemental selenium. The results also indicated that pH 7 was the optimum pH, as it displayed 27.2% selenite reduction with a net dry weight of 6.8 mg/mL. Increasing the concentration of sulfate resulted in a significant increase in selenite reduction, as it reached a maximum value of 75.3% at 0.15% g/ml sulfate. The maximum reduction in sodium selenite content was 85.2% at a C/N ratio of 2:1. The biosynthesized SeNPs exhibited antifungal activity against several fungi, such as Aspergillus flavus, Aspergillus niger, and Fusarium oxysporum, that were isolated from animal and poultry feed. Elevated SeNP concentrations (10500 ppm) significantly inhibited fungal growth. SeNPs at a concentration of 5000 ppm inhibited aflatoxin production (B1, B2, G1, and G2) by A. flavus, in addition to inhibiting mycotoxin production (T2 toxin, fumonisin B1, zearaleone, fusarin C, and moniliformin) by F. oxysporum. In conclusion, the results revealed favorable nutritional conditions for the maximum production of SeNPs by Fusarium oxysporum (CCASU-2023-F9) and indicated the marked inhibitory effect of SeNPs on mycotoxins that contaminate animal feed, causing serious consequences for animal health, and that lead to improving the quality of commercially produced animal feed. The obtained results can serve as a basis for commercial applicability.

Guide DNA dephosphorylation-modulated Pyrococcus furiosus Argonaute fluorescence biosensor for the detection of alkaline phosphatase and aflatoxins B1.

Foodborne hazardous factors pose a significant risk to public health, emphasizing the need for the development of sensitive and user-friendly detection strategies to effectively manage and control these risks in the food supply chain. Pyrococcus furiosus argonaute (PfAgo)-based biosensing approaches have been extensively explored due to its built-in signal amplification. However, the property that PfAgo is a DNA-guided DNA endonuclease has enabled almost all the existing PfAgo-based reports to be used for the detection of nucleic acids. To lend PfAgo toolbox to extended non-nucleic acid detection, we systematically investigated the mechanism characteristic of PfAgo' preference for guide DNA (gDNA) and proposed a gDNA dephosphorylation-modulated PfAgo sensor for the detection of non-nucleic acid targets. Our results indicated that PfAgo exhibits preference for 5'-phosphorylated gDNA at a specific ratio of PfAgo to gDNA concentration. Leveraging this PfAgo' preference and the dephosphorylation activity of alkaline phosphatase (ALP), ALP could be detected as low as 2.7 U/L. Furthermore, the PfAgo was coupled with immunolabelled ALP to develop a PfAgo-based fluorescence immunosensor, which achieves aflatoxins B1 detection with a detection limit of 29.89 pg/mL and exhibits satisfactory recoveries in wheat and maize samples. The developed method broadens the application scope of PfAgo toolbox, and provides a simple, sensitive, and universal detection platform for a variety targets.

Mycotoxins and coccidiosis in poultry - co-occurrence, interaction, and effects.

Avian coccidiosis, a common disease caused by Eimeria species, results in significant losses in global poultry production. Mycotoxins are low-molecular-weight natural products (i.e., small molecules) produced as secondary metabolites by filamentous fungi and they have the potential to economically and significantly affect global poultry production. Little is known about the relationship between mycotoxins and avian coccidiosis, although they often co-occur in the field. This comprehensive review examines the intricate relationship between mycotoxins and avian coccidiosis, in particular how mycotoxins, including aflatoxins, ochratoxins, trichothecenes as well as Fusarium mycotoxins, compromise the health of the poultry flock and open the door to Eimeria parasites in the gut. In addition, this review sheds light on the immunosuppressive effects of mycotoxins, their disruption of cellular signaling pathways, and the consequent exacerbation of coccidiosis infections. The mechanisms of mycotoxin toxicity are also reviewed, emphasizing direct damage to intestinal epithelial cells, impaired nutrient absorption, inflammation, oxidative stress, and changes in the gut microbiota. Finally, the consequences for the prevention and treatment of coccidiosis when mycotoxins are present in the feed are discussed. This review emphasizes the need for effective management strategies to mitigate the combined risks of mycotoxins and coccidiosis and highlights the complexity of diagnosing and controlling these interrelated problems in poultry. The review advocates a holistic approach that includes strict feed management, disease prevention measures and regular monitoring to maintain the health and productivity of poultry against these significant challenges.

Processing complementary foods to reduce mycotoxins in a medium scale Tanzanian mill: A hazard analysis critical control point (HACCP) approach.

Designing and implementing processing procedures for producing safe complementary foods in dynamic and unregulated food systems where common food staples are frequently contaminated with mycotoxins is challenging. This paper presents lessons about minimizing aflatoxins (AF) in groundnut flour and AF and/or fumonisins (FUM) in maize and groundnut pre-blended flour for complementary feeding in the context of a dietary research intervention in rural Tanzania. The flours were processed in collaboration with Halisi Products Limited (Halisi), a medium scale enterprise with experience in milling cereal-based flours in Arusha, Tanzania. Using a hazard analysis critical control point (HACCP) approach for quality assurance, two critical control points (CCPs) for AF in processing the pre-blended flour were identified: 1) screening maize before procurement, and 2) blending during the processing of each constituent flour. Blending of maize flour was also identified as a CCP for FUM. Visual inspection during screening and sorting were identified as important control measures for reducing AF, but these steps did not meet the criteria for a CCP due to lack of objective measurement and verifiable standards for AF. The HACCP approach enabled the production of low AF (<5 µg/kg) and FUM (<2 µg/g) flours with low rejection rates for the final products. The paper presents practical lessons that could be of value to a range of commercial processors in similar low- and middle-income contexts who are keen on improving food quality.

Potential Impact of Current Agricultural Practices on Mycotoxin Occurrence and Mycotoxin Knowledge Along the Cassava Value Chain in Uganda.

Cassava is the second most important staple food crop for Uganda and is prone to contamination with mycotoxins. This study aimed at understanding the current agricultural practices, their potential influence on mycotoxin occurrence, as well as assessing mycotoxin knowledge among key cassava value chain actors, including farmers, wholesalers, and processors. Data were collected through individual interviews (210), key informant interviews (34), and 4 focus group discussions. The findings revealed that 51% of farmers peeled cassava directly on bare ground, resulting in direct contact with soil that potentially harbors mycotoxin-producing fungi, such as Aspergillus section Flavi. During postharvest handling, 51.6% of farmers dried cassava chips directly on bare ground. Nearly, all (95.2%) of wholesalers packed cassava chips in local gunny bags and placed them on ground instead of pallets. In the processing of cassava chips into flour, only one of the 14 processing machines was certified by the Uganda National Bureau of Standards. Additionally, there was only one processing machine available for every 180 (1:180) consumers bringing their cassava for processing. 50.8% of cassava consumers interviewed admitted to consuming cassava flour regardless of quality, while 73% blended cassava flour with flour from mycotoxin-susceptible crops mainly maize, millet, and sorghum. Most (96.2%) of the people along the cassava value chain did not understand what the term mycotoxins meant. However, 56% of interviewed respondents were familiar with the term aflatoxins. Of the cassava value chain actors aware of mycotoxins, 82.9% knew of methods for reducing aflatoxin contamination, but only 40.9% were putting such methods into practice. More farmers (47.9%) managed aflatoxins compared to wholesalers (33.3%) and processors (21.4%). Knowledge on aflatoxins was significantly associated with value chain actor (P = 0.026), head of household (P = 0.004), region (P = 0.033), age (P = 0.001), and experience (P = 0.001). This study highlights the critical areas of mycotoxin contamination within the cassava value chain in Uganda and underscores the need to improve the knowledge among value chain actors especially farmers.

Analysis and evaluation of in vitro bioaccessibility of aflatoxins B1, B2, G1 and G2 in plant-based milks.

Plant-based milks emerge as a healthy and vegan alternative for human diet, but these foodstuffs are susceptible to be contaminated by aflatoxins. A new method based on SPE and HPLC-MS/MS analysis was optimized and validated to test the presence of aflatoxins B1, B2, G1 and G2 analysis in almond, oat, rice and soy commercial milks. Moreover, aflatoxin bioaccessibility was evaluated through an in vitro digestion assay applied to each type of spiked milk. Aflatoxins B1, B2 and G1 were detected in one soy milk sample below the LOQ, fulfilling the limits stablished by the European Legislation. The final bioaccessibility percentages were highly dependent on the type of mycotoxin and sample matrix, the highest and the lowest values were obtained for AFB2 (82%-92%) and AFG1 (15%-30%), whereas AFB1 (28%-50%) and AFG2 (32%-76%) values resulted more influenced by the plant-based milk matrix.