Mycobiota contaminating some market cake samples with reference to their toxin and enzyme.

Fungi can spoil the majority of baked products. Spoilage of cake during storage is commonly associated with fungi. Therefore, this study aimed to assess the quality of different types of cakes sold in the market. The most predominant fungal genera in the tested cake samples (14 samples) were Aspergillus spp., and Penicillium spp. On Potato Dextrose Agar (PDA), the medium fungal total count was 43.3 colonies /g. Aspergillus was the most dominant genus and was isolated from six samples of cake. Aspergillus was represented by 3 species namely, A. flavus, A. niger, and A. nidulans, represented by 13.32, 19.99, and 3.33 colonies /g respectively. On Malt Extract Agar (MEA) Medium, the fungal total count was 123.24 colonies / g. Aspergillus was the most dominant isolated genus from 11 samples of cake and was represented by 5 species, namely, A. flavus, A. niger, A. ochraceous, A. terreus, and A. versicolor (26. 65, 63.29, 3.33, 6.66, and 3.33 colonies / g , respectively). Twenty-four isolates (88.88 %) of the total tested twenty-seven filamentous fungi showed positive results for amylase production. Ten isolates (37.03%) of the total tested filamentous fungi showed positive results for lipase production, and finally eleven isolates (40.74 %) of the total fungal isolates showed positive results for protease production. Aflatoxins B1, B2, G1, G2, and ochratoxin A were not detected in fourteen collected samples of cake. In this study, clove oil was the best choice overpeppermint oil and olive oil for preventing mold development when natural agents were compared. It might be due to the presence of a varietyof bioactive chemical compounds in clove oil, whose major bioactive component is eugenol, which acts as an antifungal reagent. Therefore, freshly baked cake should be consumed within afew days to avoid individuals experiencing foodborne illnesses.

Mycotoxin detection in selected medicinal plants using chromatographic techniques.

Mycotoxins are toxic mycological products that when consumed, absorbed or inhaled cause sickness or even the death of humans. Therefore, the present study aimed to evaluate the contamination levels of mycotoxins (aflatoxins, AFB1 , AFB2 , AFG1 , AFG2 , and ochratoxin A, OTA) in selected medicinal herbs and shrubs using thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). A total of 15 samples of medicinal herbs and shrubs were selected. Among them, four samples were aflatoxin contaminated while two samples were ochratoxin A contaminated. The highest level of aflatoxin was detected in Justicia adhathoda (4,704.94 ppb) through HPLC (153.4 ppb) and through TLC, while the lowest level of aflatoxin was detected in Pegnum harmala (205.1 ppb) through HPLC. Similarly, the highest level of OTA was detected in Dodonia viscosa (0.53 ppb) through HPLC (0.5 ppb) and through TLC, while the lowest level was detected in J. adhathoda (O.11 ppb) through HPLC (0.4 ppb) and through TLC. The OTA concentration was very low, being negligible and below permissible limits. The present study concludes that there is a potential risk for the consumption of herbal decoctions. Therefore, regular monitoring and proper management of mycotoxins, including aflatoxins and OTA, in herbal medicines are needed to ensure the safety of herbal drugs to protect consumers.

Phytochemical-mediated regulation of aflatoxigenic fungi contamination in a shifting climate and environment.

Mycotoxin contamination poses a significant problem in developing countries, particularly in northern Pakistan's fluctuating climate. This study aimed to assess aflatoxin contamination in medicinal and condiment plants in Upper Dir (dry-temperate) and Upper Swat (moist-temperate) districts. Plant samples were collected and screened for mycotoxins (Aflatoxin-B1 and Aflatoxin-B-2). Results showed high levels of AFB-1 (11,505.42 ± 188.82) as compared to AFB-2 (846 ± 241.56). The maximum contamination of AFB-1 in Coriandrum sativum (1154.5 ± 13.43 ng to 3328 ± 9.9 ng) followed by F. vulgare (883 ± 9.89 ng to 2483 ± 8.4 ng), T. ammi (815 ± 11.31 ng to 2316 ± 7.1 ng), and C. longa (935.5 ± 2.12 ng to 2009 ± 4.2 ng) while the minimum was reported in C. cyminum (671 ± 9.91 ng to 1995 ± 5.7 ng). Antifungal tests indicated potential resistance in certain plant species (C. cyminum) while A. flavus as the most toxins contributing species due to high resistance below 80% (54.2 ± 0.55 to 79.5 ± 2.02). HPLC analysis revealed hydroxyl benzoic acid (5136 amu) as the dominant average phytochemical followed by phloroglucinol (4144.31 amu) with individual contribution of 8542.08 amu and 12,181.5 amu from C. cyaminum. The comparison of average phytochemicals revealed the maximum concentration in C. cyminum (2885.95) followed by C. longa (1892.73). The findings revealed a statistically significant and robust negative correlation (y = - 2.7239 × + 5141.9; r = - 0.8136; p < 0.05) between average mycotoxins and phytochemical concentrations. Temperature positively correlated with aflatoxin levels (p < 0.01), while humidity had a weaker correlation. Elevation showed a negative correlation (p < 0.05), while geographical factors (latitude and longitude) had mixed correlations (p < 0.05). Specific regions exhibited increasing aflatoxin trends due to climatic and geographic factors.

Preharvest insect pests of peanuts and associated aflatoxin contaminants in Georgia, USA.

On-farm losses of peanuts (Arachis hypogaea L., Fabales: Fabaceae) pose a persistent threat to the sustainable production and value of peanuts in the United States. This study presents empirical data on the spatial distribution of subterranean insect pests and various quality aspects of peanuts. Surveys were conducted in 20 randomly selected peanut fields in 10 counties in Northeast, Southeast, and Southwest Georgia. The primary insect pests found in Georgia's peanut production counties were Pangaeus bilineatus (Say), Elasmopalpus lignosellus (Zeller), and Diabrotica undecimpunctata Howardi. In the northeast counties, a high prevalence of P. bilineatus led to a significant increase in insect-damaged pods (%IDP), insect-damaged kernels (%IDK), discolored kernels (%DK), pod weight loss (%PWL), and kernel weight loss (%KWL). Similarly, southeast counties had a high %DK, cracked pods (%CP), and E. lignosellus infestation. In southwest counties, predominantly high D. undecimpunctata infestations resulted in the highest %IDP. Moisture content (%MC) was excessively high in all the counties (22.19%-23.17%). Preharvest aflatoxin contamination in peanuts was prevalent across all studied locations, particularly in counties with a high incidence of P. bilineatus and may cause increased risk in aflatoxin levels along the supply chain. Nevertheless, the diverse regional abundance of insect pests and the widespread presence of aflatoxins in Georgia's peanut fields offer valuable insights for developing integrated pest management strategies targeting subterranean insect pests. This is especially crucial in addressing the impact of P. bilineatus, E. lignosellus, and D. undecimpunctata on aflatoxins content of peanuts and determining the pathway for mitigation of aflatoxin contaminations in peanuts at harvest.

Guest adaptative supramolecular sensing strategy for warning the risky aflatoxins in contaminated cereals.

In the face of diversified analytes, it is a great challenge and infeasible task to design and synthesize corresponding macrocyclic hosts to realize the ideal supramolecular sensing. Herein, we proposed a novel supramolecular sensing strategy, guest adaptative assay (GAA), in which analyte was quantitatively transformed under mild conditions to perfectly adapt to macrocyclic host. As a health-threatening "landmine" in cereals, aflatoxins were converted by the aid of alkali hydrolysis to satisfactorily obtain aflatoxins transformants in ionic state, resulting in sensitive response by the guanidinocalix[5]arene•fluorescein reporter pair. Surprisingly, the established strategy not only exhibited effective practicality in screening out high-risk cereals contaminated with aflatoxins, but also relieved the laborious task of macrocycle design and screening in supramolecular sensing.

Cu-nanoparticles@ graphene nanocomposite: A robust and efficient nanocomposite for micro-solid phase extraction of trace aflatoxins in different foodstuffs.

Cu-nanoparticles-immobilized graphene (Cu@G) nanocomposite was fabricated in this study by reducing Cu(II) ions in the presence of graphene oxide using a simple chemical reduction step. Cu@G nanocomposite was applied as a sorbent for the SPE of four aflatoxins (AFs). A reusable syringe was filled with the fabricated nanocomposite and used as a sorbent for the micro-solid phase extraction of four AFs (AFB1, AFB2, AFG1, AFG2). The impact of different analytical factors was fully investigated and optimized. Excellent recoveries, ranging from 92.0 to 108.5 %, were detected when evaluating target AFs in samples of rice, maize, and pistachio. The LOD, LOQ, and linear ranges were attained under optimal circumstances in the ranges of 0.0062 µg kg-1, 0.0192 µg kg-1, and 0.0-20 µg kg-1, respectively. The discovered approach provided the dual benefits of a high enrichment capability of Cu-nanoparticles via AFs complexation and a huge porosity of graphene sheets.

The impact of fullerenol nanoparticles on the growth of toxigenic Aspergillus flavus and aflatoxins production in vitro and in corn flour.

Antifungal and antimycotoxigenic activity of fullerenol nanoparticles (FNPs) were investigated on Aspergillus flavus growth isolated from a real food sample and aflatoxins (AFs) (AFB1 and AFB2 ) production. The final FNPs concentrations in in vitro and in commercial corn flour after the stationary incubation period of 7 and 14 days were in the range 0.16-80 µg/mL and 0.16-80 µg/g, respectively. Nanocharacterization of FNPs revealed an average size of 5-20 nm and a zeta potential of -35 mV. The highest degree of A. flavus mycelium growth inhibition (28%) after 7 days was observed for applied FNP concentration of 8.0 µg/mL, while after 14 days FNP concentration of 0.32 µg/mL led to the maximal inhibition of A. flavus mycelium growth (36%). Spearman's correlations analysis revealed a strong positive correlation between AFB1 and AFB2 concentrations in YES broth after 7 (R = 0.994, p < 0.05) and 14 days (R = 0.976), as well as between AFs concentrations and A. flavus mycelium mass after 7 (R = 0.786 for AFB1 and R = 0.766 for AFB2 ) and 14 days (R = 0.810 for AFB1 and R = 0.833 for AFB2 ). Paired samples t-test showed the existence of a statistically significant difference (p < 0.05) between the produced AFs concentrations after the incubation of 7 and 14 days. Regarding the artificially inoculated corn flour the lower applied FNP concentrations (0.16-0.8 µg/g) achieved a reduction of AFB1 up to 42% and 60% after 7 and 14 days, respectively.

Metabolites and degradation pathways of microbial detoxification of aflatoxins: a review.

The degradation of aflatoxins using nonpathogenic microbes and their enzymes is emerging as a safe and economical alternative to chemical and physical methods for the detoxification of aflatoxins in food and feeds. Many bacteria and fungi have been identified as aflatoxin degraders. This review is focused on the chemical identification of microbial degradation products and their degradation pathways. The microbial degradations of aflatoxins are initiated by oxidation, hydroxylation, reduction, or elimination reactions mostly catalyzed by various enzymes belonging to the classes of laccase, reductases, and peroxidases. The resulting products with lesser chemical stability further undergo various reactions to form low molecular weight products. Studies on the chemical and biological nature of degraded products of aflatoxins are necessary to ensure the safety of the decontamination process. This review indicated the need for an integrated approach including decontamination studies using culture media and food matrices, proper identification and toxicity profiling of degraded products of aflatoxins, and interactions of microbes and the degradation products with food matrices for developing practical and effective microbial detoxification process.

Characterization of Aspergillus section Flavi associated with stored grains.

Increased frequencies of Aspergillus section Flavi and aflatoxins in cereal grains have been seen in recent years due to changes in climate circumstances, such as high temperatures and drought. To assess the microbiological risks of contamination, it is critical to have a reliable and accurate means of identifying the fungi. The main goal of this study was to characterize Aspergillus species from section Flavi obtained from twenty-three samples of barley and maize grains, gathered from different markets in Qena, Egypt, using morphological and molecular techniques. Twenty-three isolates were chosen, one isolate from each sample; they were identified as A. aflatoxiformans (4 isolates), A. flavus (18), and A. parasiticus (1). The existence of four aflatoxin biosynthesis genes was also investigated in relation to the strains' ability to produce total aflatoxins and aflatoxin B1, focusing on the regulatory gene aflR and the structural genes aflD and aflM. All strains producing aflatoxins were linked to the presence of aflR1 and/or aflR2, except two isolates that exhibited aflatoxins but from which aflR1 or aflR2 were not detected, which may be due to one or more missing or unstudied additional genes involved in aflatoxin production. AflD and aflM genes were amplified by 10 and 9 isolates, respectively. Five samples of barley and maize were contaminated by aflatoxins. Fifteen isolates were positive for producing total aflatoxins in the range of 0.1-240 ppm. Antagonistic activity of Trichoderma viride against A. flavus (F5) was assessed at 31.3%. Trichoderma reduced total aflatoxins in all treated seeds, particularly those subjected to Trichoderma formulation.

Adsorption of aflatoxin B1 to corn by-products.

The adsorption of aflatoxin B1 (AFB1) to natural fiber materials prepared from corn by-products was investigated in this study. The results showed that corn cob powder (CCP) dose, particle size, time (0.25-24 h), temperature (4, 20, 37, 50 and 100 °C) and pH (2-8), had significant effects on adsorption. The maximum adsorption (98%) was with particles 500-355 µm in size at 20 °C for 8 h, at the dose of 50 mg mL-1. The adsorption fitted pseudo-second-order model and Langmuir isotherm well. Besides, CCP had a higher adsorption capacity to AFB1 than any single cell wall components of corn, which indicated that capillary effect happened in cell wall might be the main reason for adsorption. The results also suggested that CCP could reduce AFB1 content from both liquid and solid food matrixes. Briefly, CCP displayed promising properties that could be developed in nature-based practical applications for food aflatoxin decontamination.

The potential for reducing aflatoxin B1 contamination of stored peanuts by soil disinfection.

Aflatoxins from the fungus Aspergillus flavus (A. flavus) that contaminate stored peanuts is a major hazard to human health worldwide. Reducing A. flavus in soil can decrease the risk of aflatoxins in stored peanuts. In this experiment, we determined whether peanuts grown on soil fumigated with dazomet (DZ), metham sodium (MS), allyl isothiocyanate (AITC), chloropicrin (PIC) or dimethyl disulfide (DMDS) would reduce of the quantity of A. flavus and its toxin's presence. The results of bioassays and field tests showed that PIC was the most effective fumigant for preventing and controlling A. flavus, followed by MS. PIC and MS applied to the soil for 14 d resulted in LD50 values against A. flavus of 3.558 and 4.893 mg kg-1, respectively, leading to almost 100% and 98.82% effectiveness of A. flavus, respectively. Peanuts harvested from fumigated soil and then stored for 60 d resulted in undetectable levels of aflatoxin B1 (AFB1) compared to unfumigated soil that contained 0.64 ug kg-1 of AFB1, which suggested that soil fumigation can reduce the probability of aflatoxin contamination during peanut storage and showed the potential to increase the safety of peanuts consumed by humans. Further research is planned to determine the practical value of our research in commercial practice.

Validation of the AgraStrip® Pro Total Aflatoxin WATEX® Method for Detection of Total Aflatoxins in Corn and Peanut Paste: AOAC Performance Tested MethodSM 032402.

BACKGROUND: To protect public and animal health against risks provoked by aflatoxins contained therein, maximum limits for aflatoxins are defined. Limit values vary depending on the intended use and regulatory authority, therefore quantitative detection is essential. OBJECTIVE: Validation of a one-step competitive lateral flow immunochromatographic assay for quantitative screening of total aflatoxin (B1, B2, G1, and G2) in corn and peanut paste for the high-sensitivity range (0-50 µg/kg). METHODS: Corn or peanut paste test portions are water-based extracted and prepared for testing within 15 min. The AgraStrip® Pro Total Aflatoxin WATEX® test method quantifies the concentration of aflatoxins in the sample. Selectivity, robustness, product consistency, and stability testing were performed in addition to matrix testing. RESULTS: No cross-reactivity was detected against possible interferants. Corn resulted in a LOD and LOQ of 0.9 and 2.8 µg/kg and overall recoveries between 74 and 108%. Peanut paste resulted internally in a LOD and LOQ of 0.8 and 2.3 µg/kg and recoveries between 86 and 98%. Stability testing showed no influence of the age of the respective lot on the result. Robustness testing demonstrated that varying the amount of water used for extraction, extraction time, and delay between extract dilution and analysis did not significantly affect the result. Due to supply chain issues, a change to the outer cartridge required an increase in the test aliquot size, which had no effect on method performance. CONCLUSION: The test kit was validated for the determination of total aflatoxins in corn and peanut paste. Recovery and precision met the requirements laid down in Codex Alimentarius CXG 71-2009 and acceptable robustness, selectivity, and product consistency and stability were demonstrated. HIGHLIGHTS: The AgraStrip Pro Total Aflatoxin WATEX test kit in the high sensitivity range (0-50 µg/kg) was approved by the AOAC AOAC Research Institute (PTM number 032402).

Aflatoxins and fumonisins in conventional and organic corn: a comprehensive review.

This review analyzes the occurrence and co-exposure of aflatoxins and fumonisins in conventional and organic corn, and compares the vulnerability to contamination of both. The risks of fungal contamination in corn are real, mainly by the genera Aspergillus and Fusarium, producers of aflatoxins and fumonisins, respectively. Aflatoxins, especially AFB1, are related to a high incidence of liver cancer, and the International Agency Research of Cancer (IARC) classified them in group 1A 'carcinogenic to humans'. The occurrence in conventional corn is reported in many countries, including at higher levels than those established by legislation. IARC classified fumonisins in group 2B 'possibly carcinogenic to humans' due to their link with incidence of esophageal cancer. However, comparing corn and organic and conventional by-products from different regions, different results are observed. The co-occurrence of both mycotoxins is a worldwide problem; nevertheless, there is little data on the comparison of the co-exposure of these mycotoxins in corn and derivatives between both systems. It was found that the agricultural system is not a decisive factor in the final contamination, indicating the necessity of effective strategies to reduce contamination and co-exposure at levels that do not pose health risks.

Evaluation of Aflatoxins Occurrence and Exposure in Cereal-Based Baby Foods: An Update Review.

PURPOSE OF REVIEW: The first stages of human life, which include the fetal period, infancy, and early childhood, are the most critical for human growth and development. This is the most vulnerable phase to health challenges due to the immature immune system and rapid development. Mycotoxins such as aflatoxins, ochratoxin A, patulin, fumonisins, zearalenone, and deoxynivalenol are secondary metabolites secreted by various fungal species, primarily Aspergillus, Fusarium, Penicillium, and Alternaria. Aflatoxins are one of the major mycotoxins produced in cereals and cereal-based foods by several species of Aspergillus, mainly Aspergillus flavus. In this context, this review provides a brief overview of the occurrence, exposure, legal regulations, and health effects of aflatoxins (B1, B2, G1, G2, and M1) in cereal-based baby foods and breast milk. RECENT FINDINGS: Human aflatoxin exposure in utero and through breast milk, infant formulas, cereals, and cereal-based foods has been linked to various health consequences, including adverse birth outcomes, impaired growth and development, immune system suppression, and hepatic dysfunction. Recent evidence suggests that especially infants and children are more susceptible to aflatoxins due to their lower body weight, lowered capacity to detoxify harmful substances, more restrictive diet, immature metabolism and elimination, and faster rates of growth and development. It is essential for both food safety and infant and child health that aflatoxins in cereal and cereal-based products are precisely detected, detoxified, and managed.

The occurrence of aflatoxins and labelling compliance of locally produced za'atar mix products.

Za'atar mix products are mainly composed of the dried and ground leaves and/or blossoms of wild and cultivated plant species (Origanum, Thymbra, Thymus, and Satureja) with the addition of condiments. The aim of this study was to evaluate the occurrence of aflatoxins, chemical composition (carbohydrates, fibre, fat, protein, moisture, ash, and acid contents), mineral content (Na, Ca, and K), and colour traits (L*a*b*) in relation to food label and food standards compliance. Measured and labelled fat content did not agree for approximately 91% of the samples. There was also no agreement between the measured and labelled fibre contents. The total content of aflatoxins in the tested samples ranged from 2 to 63.7 ng g-1. Eleven (69%) of the 16 analysed products had total aflatoxins higher than the maximum permitted limit of the European Commission. The KAS and LAZ products had significantly lighter colour (the highest L* values), while the ALAQ product had the darkest colour (lowest L* value). The range of sodium content in the tested products was 105.1-1425.3 mg/100 g. In conclusion, za'atar mix products that are available in local markets do not have accurate nutritional labelling information, and the occurrence of aflatoxins was very high. Further studies are needed to evaluate the reasons for these quality defects.

Enhanced detection of Aspergillus flavus in peanut kernels using a multi-scale attention transformer (MSAT): Advancements in food safety and contamination analysis.

In this study, a multi-scale attention transformer (MSAT) was coupled with hyperspectral imaging for classifying peanut kernels contaminated with diverse Aspergillus flavus fungi. The results underscored that the MSAT significantly outperformed classic deep learning models, due to its sophisticated multi-scale attention mechanism which enhanced its classification capabilities. The multi-scale attention mechanism was utilized by employing several multi-head attention layers to focus on both fine-scale and broad-scale features. It also integrated a series of scale processing layers to capture features at different resolutions and incorporated a self-attention mechanism to integrate information across different levels. The MSAT model achieved outstanding performance in different classification tasks, particularly in distinguishing healthy peanut kernels from those contaminated with aflatoxigenic fungi, with test accuracy achieving 98.42±0.22%. However, it faced challenges in differentiating peanut kernels contaminated with aflatoxigenic fungi from those with non-aflatoxigenic contamination. Visualization of attention weights explicitly revealed that the MSAT model's multi-scale attention mechanism progressively refined its focus from broad spatial-spectral features to more specialized signatures. Overall, the MSAT model's advanced processing capabilities marked a notable advancement in the field of food quality safety, offering a robust and reliable tool for the rapid and accurate detection of Aspergillus flavus contaminations in food.

Fluorine-functionalized covalent organic framework as efficient solid phase extraction sorbent for adsorption of aflatoxins in nuts.

In this study, a new fluorine-functionalized covalent organic framework (F-COF) was designed and fabricated by the direct polycondensation of tris(4-aminophenyl)amine and 2,3,5,6-tetra-fluoroterephthaldehyde for the first time. F-COF exhibited a remarkably enhanced adsorption capability compared with that of the fluorine-free COF. The favorable adsorption of aflatoxins was attributed to multiple interactions including pseudo hydrogen bond, F-O, π-π, F-π interactions and hydrophobic interactions between F-COF and aflatoxins. By coupling F-COF based solid phase extraction with high-performance liquid chromatography equipped with fluorescence detector, a rapid and sensitive method for determining aflatoxins (aflatoxin B1, B2, G1 and G2) in nuts (peanuts and pistachios) was established. Under optimal conditions (35 mg F-COF, 100 mL sample solution, 3 mL min-1 as sample loading rate, pH<7, 0.2 mL acetonitrile as desorption solvent), the limits of detection for aflatoxins were 0.02-0.30 ng g-1. The linear range was 0.08-16.0 ng g-1 and the recoveries of the F-COF-based method were 83.5-114 % with relative standard deviations less than 8.0 %.

Prevalence and concentration of mycotoxins in bovine feed and feed components: A global systematic review and meta-analysis.

Mycotoxins are secondary metabolites produced by fungi and identified as contaminants in animal feed. They have potentially harmful effects, including carcinogenicity, mutagenicity, and repro-toxicity in animals and humans. As a result of climate change, there is the potential for a change in the prevalence and concentration of mycotoxins in animal feed components. This necessitates an assessment of the present and emerging threats to the food supply chain from mycotoxins. This systematic review and meta-analysis study synthesised studies on mycotoxin contamination and prevalence in cattle feed components. The studies were collected from scientific databases Web of Knowledge, Scopus, and Embase between 2011 and 2022. The meta-analysis synthesised 97 studies on the prevalence and the concentration of aflatoxins, ochratoxin A, deoxynivalenol, zearalenone, fumonisin and T-2/HT-2 toxins in feed components. Aflatoxin was highly prevalent (59 %), with a concentration of 2.58-3.92 µg kg-1 in feed components. Ochratoxin A had a global prevalence of 31 % with a concentration of 5.56-12.41 µg kg-1. Deoxynivalenol had a global concentration of 233.17-327.73 µg kg-1 and a prevalence of 74 %. Zearalenone had a prevalence of 70 % and a concentration of 42.47-66.19 µg kg-1. The concentration and prevalence of fumonisins was 232.19-393.07 µg kg-1 and 65 %, respectively. The prevalence and concentration of T-2/HT-2 toxins were 45 % and 23.54-35.12 µg kg-1, respectively. The synthesised concentration of the mycotoxins in the overall feed components was lower than the regulated and guidance values set by the European Union. However, in a few cases, the 95th percentile exceeded these concentration values due to high levels of uncertainty attributed to lower sample size, and thus, need to be considered while conducting risk assessments. The study highlights climates and regions likely to be conducive to the emergence of mycotoxin risk, especially considering the potential influences of climate change.

Aspergillus population diversity and its role in aflatoxin contamination of cashew nuts from coastal Kenya.

Cashew nuts are among the main cash crops in coastal Kenya, due in large part to their high nutritional value. Unfortunately, they also make them highly susceptible to mold contamination, resulting in biodeterioration of the nutritional value and potential contamination with toxic secondary metabolites, such as aflatoxins, that cause them to be rejected for sale at the market. We determined the population diversity of the Aspergillus species and their role in aflatoxin contamination in cashew nuts in selected coastal regions of Kenya. Fifty raw cashew nut samples were collected from post-harvest storage facilities across three counties in Kenya's coastal region and examined for moisture content and the presence of Aspergillus fungi. About 63 presumptive isolates were recovered from the cashew nuts. ITS and 28S rDNA regions were sequenced. The aflD, aflM and aflR genes were amplified to identify the potentially aflatoxigenic from the Aspergillus isolates. The Aflatoxins' presence on the isolates was screened using UV and the ammonia vapour test on coconut milk agar and validated using ELISA assay. A comparison of cashew moisture content between the three counties sampled revealed a significant difference. Sixty-three isolates were recovered and identified to section based on morphological characters and their respective ITS regions were used to obtain species identifications. Three sections from the genus were represented, Flavi and Nigri, and Terrei with isolates from the section Nigri having slightly greater abundance (n = 35). The aflD, aflM and aflR genes were amplified for all isolates to assess the presence of the aflatoxin biosynthesis pathway, indicating the potential for aflatoxin production. Less than half of the Aspergillus isolates (39.68%) contained the aflatoxin pathway genes, while 22.22% isolates were aflatoxigenic, which included only the section Flavi isolates. Section Flavi isolates identification was confirmed by calmodulin gene. The presence of species from Aspergillus section Flavi and section Nigri indicate the potential for aflatoxin or ochratoxin in the cashew nuts. The study established a foundation for future investigations of the fungi and mycotoxins contaminating cashew nuts in Kenya, which necessitates developing strategies to prevent infection by mycotoxigenic fungi, especially during the storage and processing phases.

Determination of aflatoxins in rice from Penang, Malaysia by dispersive liquid-liquid micro-extraction and LC-MS/MS.

Rice is one of the crops cultivated in Malaysia, and it is the main diet for most of the population. In this study, dispersive liquid-liquid micro-extraction (DLLME) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to develop, optimise and validate a reliable, easy-to-use and quick approach to detect aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1) and aflatoxin G2 (AFG2). The extraction recoveries in DLLME were enhanced by the addition of 5% salt, utilising chloroform as the extraction solvent and acetonitrile as the dispersive solvent. The DLLME parameters - the extraction solvent volume, salt concentration and dispersive solvent volume were optimised with Box-Behnken design (BBD) and response surface methodology (RSM). Under optimised experimental conditions, excellent linearity was obtained with a limit of detection (LOD) ranging from 0.125 to 0.25 ng g-1, a limit of quantitation (LOQ) ranging from 0.25 to 0.3 ng g-1 and a correlation value (R2) of 0.990. The matrix effects were between -11.1% and 19.9%, and recoveries ranged from 87.4% to 117.3%. The optimised and validated method was used effectively to assess aflatoxins contamination in 20 commercial rice samples collected from local supermarkets in Penang, Malaysia. AFB1 was detected at 0.41-0.43 ng g-1 in two rice samples, below the regulatory limit.