Comprehensive Survey of Mycotoxin Occurrence in Indian Food Commodities.

The prevalence and concentration of mycotoxins in rice sourced from markets: A global description

The spatiotemporal trends in aflatoxin B1 (AFB1), fumonisin B1 (FB1), and deoxynivalenol (DON) accumulation were analyzed in a range of food commodities (maize, groundnut, pearl millet, rice, and wheat) in village settings in Unnao, Uttar Pradesh, India. Samples (n = 1549) were collected across six communities and six time points spanning a calendar year and were analyzed for mycotoxins using enzyme-linked immunosorbent assays. AFB1 and FB1 were common across surveyed villages, with moderate to high detection rates (45–75%) observed across commodities. AFB1 levels in maize and groundnuts and FB1 levels in maize and pearl millet frequently exceeded regulatory threshold levels of 15 μg/kg (AFB1) and 2 μg/g (FB1). DON was analyzed in wheat, with 3% of samples yielding detectable levels and none exceeding 1 μg/g. In rice, AFB1 levels were highest in the bran and husk and lower in the kernel. Commodity type significantly influenced AFB1 detection status, while commodity type, season, and visual quality influenced samples’ legal status. Storage characteristics and household socioeconomic status indicators did not have significant effects on contamination. No significant effects of any variables on FB1 detection or legal status were observed. Data on mycotoxin contamination, combined with data on local dietary intake, were used to estimate spatiotemporal mycotoxin exposure profiles. Estimated seasonal per capita exposure levels for AFB1 (5.4–39.3 ng/kg body weight/day) and FB1 (~0–2.4 μg/kg body weight/day) exceeded provisional maximum tolerable daily intake levels (1 ng/kg body weight/day for AFB1 and 2 μg/kg body weight/day for FB1) in some seasons and locations. This study demonstrates substantial dietary mycotoxin exposure risk in Unnao food systems and serves as an evidentiary foundation for participatory food safety intervention in the region.

Multiplex fluorescence quenching immunoassay based on multicolor magnetic quantum dot and dual spectral-overlapped polydopamine nanospheres for ultrasensitive detection of aflatoxin B1, zearalenone, ochratoxin A, and fumonisin B1

Fumonisins in Maize: Can We Reduce Their Occurrence?

Human oral exposure to aflatoxin B1 (AFB1 ) and fumonisin B1 (FB1 ) is associated with increased hepatocellular carcinoma. Although evidence suggested interactive AFB1 -FB1 hepatotoxicity, the underlying mechanisms remain mostly unidentified. This work was aimed at evaluating the possible AFB1 -FB1 interplay to induce genetic and cell cycle toxicities in BRL-3A rat hepatocytes, reactive oxygen species (ROS) involvement, and the AFB1 metabolizing pathways cytochrome P450 (CYP) and arachidonic acid (ArAc) metabolism as ROS contributors. Flow cytometry of stained BRL-3A hepatocytes was used to study the cell cycle (propidium iodide), ROS intracellular production (DCFH-DA, HE, DAF-2 DA), and phospholipase A activity (staining with bis-BODIPY FL C11-PC). The CYP1A activity was assessed by the 7-ethoxyresorufin-O-deethylase (EROD) assay. Despite a 48-h exposure to FB1 (30μM) not being genotoxic, the AFB1 (20μM)-induced micronucleus frequency was overcome by the AFB1 -FB1 mixture (MIX), presumably showing toxin interaction. The mycotoxins blocked G1/S-phase, but only MIX caused cell death. Overall, the oxidative stress led these alterations as the pretreatment with N-acetyl-l-cysteine reduced such toxic effects. While AFB1 had a major input to the MIX pro-oxidant activity, with CYP and ArAc metabolism being ROS contributors, these pathways were not involved in the FB1 -elicited weak oxidative stress. The MIX-induced micronucleus frequency in N-acetyl-l-cysteine pretreated cells was greater than that caused by AFB1 without antioxidants, suggesting enhanced AFB1 direct genotoxicity probably owing to the higher CYP activity and ArAc metabolism found in MIX. The metabolic pathways modulation by AFB1 -FB1 mixtures could raise its hepatocarcinogenic properties.

Liquid chromatography coupled with single or tandem mass spectrometry (LC-MS/(MS)) is routinely used for the simultaneous determination of mycotoxins in food and feed although official methods using this technique have not yet been adopted by the European Committee for Standardization and the Association of Analytical Communities. A proficiency test (PT) was conducted for the simultaneous determination of up to 11 mycotoxins (aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), ochratoxin A (OTA), deoxynivalenol (DON), T-2 toxin (T-2), HT-2 toxin (HT-2), zearalenone (ZEA), fumonisin B1 (FB1) and fumonisin B2 (FB2)) in maize using LC-MS/(MS) to benchmark laboratories currently using this technique and to obtain information on currently used methodologies and method-related performances. Each participant received the following: instructions; a comprehensive questionnaire; a mixed mycotoxins calibration solution; a spiking solution (AFB1, AFB2, AFG1 and AFG2, OTA, DON, T-2, HT-2, ZEA, FB1 and FB2); and two test materials, namely a contaminated maize sample and a blank maize sample to be spiked with a spiking solution containing 11 mycotoxins. Laboratory results were rated with z-scores. Of the 64 laboratories enrolled in the PT, 41 laboratories from 14 countries returned 43 sets of results for various combinations of analytes. The majority of laboratories (61%) reported results for all 11 mycotoxins, whereas the remaining laboratories reported results for a restricted combination (from 2 to 10 analytes). For contaminated maize and spiked maize the percentage of satisfactory z-score values (|z| ?2) were: DON 55% and 49%, FB1 50% and 30%, FB2 52% and 38%, ZEA 68% and 64%, T-2+HT-2 toxins 82% and 85%, OTA 58% and 60%, AFB1 56% and 62%, AFG1 73% and 84%, AFB2 40% and 78%, AFG2 64% and 78%, respectively. The poorest performance (|z| >3) was obtained for FB1 (31%), FB2 (32%), AFB1 (32%) and AFB2 (32%) in contaminated maize and for DON (35%), FB1 (63%) and FB2 (52%) in spiked maize. Mean recovery results were acceptable for all mycotoxins (74% to 109%), except for fumonisins, where these were unacceptably high (159% for FB1 and 163% for FB2). A robust and reliable method for simultaneous determination of 11 mycotoxins in maize could not be identified from the results of this PT. Additional experimental work is necessary to set up a method suitable for inter-laboratory validation. The results of this PT and the relevant method's details can be useful to identify methodology strengths and weaknesses.

Mycotoxin contamination in poultry feed continues to pose significant challenges for animal health, food safety, and overall public health. In this study, we investigated the antifungal and antitoxigenic activities of hydro-alcoholic extracts and crude essential oils from Clove (Syzygium aromaticum), Cinnamon bark (Cinnamomum verum), Ginger (Zingiber officinale), Neem (Azadirachta indica), and Fenugreek (Trigonella foenum-graecum). These plant-derived substances were evaluated both with and without the addition of kaolin clay, targeting major mycotoxins such as Aflatoxin B1 (AFB1), Ochratoxin A (OTA), and Fumonisin B1 (FB1) levels (Table3). Our antifungal assays focused on Aspergillus flavus, A. niger, and Fusarium proliferatum. Among the tested agents, clove and cinnamon oils demonstrated the strongest antifungal properties, with clove oil providing consistent inhibition across all fungal species. Neem extracts exhibited moderate efficacy, particularly in lowering AFB1 concentrations. Notably, the incorporation of kaolin clay (1mg/g feed) enhanced FB1 detoxification, especially when combined with ginger or clove oils. In contrast, fenugreek-derived products showed minimal antifungal or antitoxigenic effectiveness. These findings highlight the potential of certain plant-based products-namely clove, cinnamon, and neem-used in conjunction with kaolin clay as sustainable alternatives to synthetic chemicals for controlling mycotoxin contamination in poultry feed. Further research is recommended to optimize dosage and application strategies to maximize their efficacy.

Fermented food samples (n = 191) including maize gruel (ogi), sorghum gruel (ogi-baba), melon seed (ogiri), locust bean (iru) and African oil bean seed (ugba) from Southwest Nigeria were quantified for 23 mycotoxins, including aflatoxin B1 (AFB1), fumonisin B1 (FB1), and sterigmatocystin (STE) using liquid chromatography-tandem mass spectrometry. The practices, perceived understanding and health risks related to fungal and mycotoxin contamination amongst fermented food sellers was also established. Data obtained revealed that 82% of the samples had mycotoxins occurring singly or in combination. FB1 was present in 83% of ogi-baba samples, whereas 20% of ugba samples contained AFB1 (range: 3 to 36 µg/kg) and STE was present in 29% of the ogi samples. In terms of multi-mycotoxin contamination, FB1 + FB2 + FB3 + STE + AFB1 + alternariol + HT-2 co-occurred within one sample. The awareness study revealed that 98% of respondents were unaware of mycotoxin contamination, and their education level slightly correlated with their level of awareness (p < 0.01, r = 0.308). The extent to which the analyzed mycotoxins contaminated these food commodities, coupled with the poor perception of the population under study on fungi and mycotoxins, justifies the need to enact fungal and mycotoxin mitigation strategies along the food chain.

Efficacy of sodium bentonite as a detoxifier of broiler feed contaminated with aflatoxin and fumonisin

Mycotoxin contamination in maize poses significant food and feed safety risks, particularly in regions with variable climatic conditions like Serbia. This study investigated the occurrence of regulated mycotoxins in maize harvested across the Republic of Serbia from 2021 to 2023, emphasizing the impact of climatic factors. A total of 548 samples of unprocessed maize grains were analysed for the presence of key mycotoxins, including aflatoxins, ochratoxin A, zearalenone, deoxynivalenol, fumonisins, and trichothecenes type A (T-2 and HT-2 toxins), using validated analytical methods. The results revealed high contamination frequencies, with aflatoxins and fumonisins being the most prevalent. The results revealed substantial temporal variability and frequent co-contamination of mycotoxins. Aflatoxin B1 (AFB1) was the most concerning contaminant, with 73.2% of the samples in 2022 exceeding the European regulatory limit for human consumption (5 µg/kg) for un processed maize grains, reaching peak concentrations of 527 µg/kg, which is 105.4 times higher than the allowed limit. For animal feed, the limit of 20 µg/kg was exceeded in 40.5% of the samples, with the highest concentration being 26.4 times greater than the maximum allowable level. In 2021, the non-compliance rates for AFB1 in food and feed were 8.3% and 2.3%, respectively, while in 2023, they were 23.2% and 12.2%, respectively. Fumonisins contamination was also high, particularly in 2021, with fumonisin B1 (FB1) detected in 87.1% of samples and average concentrations reaching 4532 µg/kg. Although levels decreased in 2023 (70.7% occurrence, average 885 µg/kg), contamination remained significant. Deoxynivalenol (DON) contamination was consistently high (>70% of samples), with peak concentrations of 606 µg/kg recorded in 2021. Zearalenone (ZEN) and ochratoxin A (OTA) occurred less frequently, but ZEN levels peaked in 2022 at 357.6 µg/kg, which is above the regulatory limit of 350 µg/kg for food. Trichothecenes (HT-2 and T-2 toxins) were detected sporadically, with concentrations well below critical thresholds. Co-occurrence of mycotoxins was frequent, with significant mixtures detected, particularly between aflatoxins and fumonisins, as well as other fusarial toxins. The analysis demonstrated that temperature, humidity, and rainfall during both the growing and harvest seasons strongly influenced mycotoxin levels, with the most severe contamination occurring under specific climatic conditions. Notably, the highest mycotoxin levels, like aflatoxins, were linked to warmer temperatures and lower rainfall. The high non-compliance rates for aflatoxins and fumonisins and co-contamination pose significant food and feed safety risks. From a public health perspective, chronic exposure to contaminated maize increases the likelihood of carcinogenesis and reproductive disorders. Reduced productivity and bioaccumulation in animal tissues/products represent serious economic and safety concerns for livestock. This study provides insights into the potential risks to food and feed safety and the need for enhanced regulatory frameworks, continuous monitoring, and mitigation strategies in Serbia as well as other geographical regions.

This study summarizes the levels of the mycotoxins aflatoxin B1, deoxynivalenol, zearalenone, ochratoxin A, and fumonisin B1 in domestic feed produced in Japan. We collected samples of Japanese domestic feed used in livestock farming establishments or by small farmers from April 2012 to March 2014, and measured mycotoxin concentrations in each sample. Regarding corn, deoxynivalenol had the highest detection rate (84%), maximum value (2370 µg/kg), and mean value (400 µg/kg). These results suggest that deoxynivalenol is a major mycotoxin contaminant in Japanese-produced domestic feed. Zearalenone and fumonisin B1 presented the second highest detection rates. The maximum concentration of zearalenone was 1200 µg/kg in grass samples, but its median was under 25 µg/kg, and, overall, it occurred at low concentrations. The detection rate of fumonisin B1 was about 30%, but its maximum concentration in corn was 2400 μg/kg, slightly higher than deoxynivalenol. Overall, mycotoxin concentrations were lower in grass than in corn. Although deoxynivalenol was detected in samples collected from all regions, concentrations in samples from Hokkaido were particularly high. Fumonisin B1 was mainly distributed in Kanto and the southern regions. Concentrations of ochratoxin A and aflatoxin B1 were low; however, the maximum concentration of aflatoxin B1 was 22 μg/kg in corn. Although aflatoxin B1 and ochratoxin A were rarely detected, they occasionally presented high levels, therefore requiring attention. Overall, mycotoxins produced by Fusarium sp. fungi require attention in Japanese-produced domestic feed.

Plant-based beverages (popularly known as vegetable milk) have become increasingly important in recent years. However, the nonexistence of information on mycotoxin contamination is noticeable. We herein describe the development and validation of an analytical methodology that employs QuEChERS and LC-MS/MS for the simultaneous determination of nine mycotoxins (aflatoxins B1, B2, G1, and G2, fumonisins B1 and B2, ochratoxin A, zearalenone, and citreoviridin) in seven types of vegetable milk (peanut, oat, rice, cashew, maize, soybean, and coconut). The method provided the following quantification limits, recoveries at the lowest validated concentration and relative standard deviations under repeatability conditions at the lowest validated concentration, respectively: aflatoxin B1 (0.023 μg/l, 84.98 and 9.23%); aflatoxin B2 (0.024 μg/, 93.00 and 4.85%); aflatoxin G1 (0.057 μg/l, 98.85 and 5.53%); aflatoxin G2 (0.031 μg/l, 96.64 and 4.08%); fumonisin B1 (2.166 μg/l, 75.55 and 16.78%); fumonisin B2 (1.105 μg/l, 70.47 and 11.89%); ochratoxin A (0.104 μg/l, 72.05 and 5.12%); zearalenone (8.093 μg/l, 107.10 and 6.37%); citreoviridin (1.305 μg/l, 97.25 and 7.28%). The method uses small amounts of samples, solvents, and other inexpensive reagents with no need for laborious clean-up and pre-concentration steps. Its attractive characteristics (simplicity, low cost compared to procedures that use immunoaffinity columns, and full compatibility with routine analyses) make it potentially valuable. As a proof-of-principle, the validated methodology was applied to seven commercial samples of different compositions showing that some were contaminated with aflatoxins and ochratoxin A.

To evaluate mycobiota and aflatoxins B(1) (AFB(1)), B(2) (AFB(2)), G(1) (AFG(1)), G(2) (AFG(2)) and fumonisin B(1) (FB(1)) contamination in different malted barley types and brands and brewer's grain collected from a major Argentinean brewery. Total fungal counts were performed using the plate count method. Aflatoxin B(1), AFB(2), AFG(1), AFG(2) and Zearalenone (ZEA) analyses were performed by thin-layer chromatography (TLC). Fumonisin B(1) was determined by HPLC. Eighty-three percentage of the malted barley (100% M1, 50% M2 and 100% M3) and 61% of brewer's grain samples had a count >1 × 10(4) CFU g(-1). Yeasts were isolated from all malt and brewer's grain samples. Genera containing some of the most important mycotoxin producer species--Fusarium ssp., Aspergillus ssp., Penicillium ssp. and Alternaria ssp.--were isolated from the analysed samples, along with other environmental saprophytic fungi such as Geotrichum ssp., Mucorales and Cladosporium ssp. All samples were contaminated with 104-145 μg kg(-1) FB(1). Eighteen per cent of brewer's grain samples were contaminated with 19-44.52 μg kg(-1) AFB(1). Aflatoxin B(2), AFG(1), AFG(2) and ZEA were not detected in any of the analysed samples. Fungal and mycotoxin contamination in malt and brewer's grain is an actual risk for animal and human health. This study may be useful for assessing the risk of mycotoxins in Argentinean beers and especially in animal feeds.

Aflatoxins and fumonisins are ubiquitous foodborne toxicants and the co-occurrence of these mycotoxins in human foods represents a significant public health concern, which has been strongly associated with human aflatoxicosis, neural tube defects, as well as many types of primary cancers. In this study the co-contamination of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) in food and human dietary exposure was investigated in residents of three different areas of China. A total of 209 food samples were measured for AFB1 and FB1. The median AFB1 levels were 13.5, 2.3 and 1.3 µg kg−1 and the median FB1 levels were 2.6, 0.4 and 0.3 mg kg−1 in corn samples collected from Huaian (a high-risk area for oesophageal cancer), Fusui (a high-risk area for liver cancer) and Huantai (a low-risk area for both oesophageal and liver cancers), respectively. The median level of AFB1 in plant oil of Fusui was the highest (52.3 µg kg−1) among all food samples analysed. Co-contamination of these two mycotoxins was found in corn, rice and wheat flour. Based on measured food consumption data, the averaged daily dietary intake of AFB1 was 0.397 µg (range = 0.269–1.218 µg) in residents of Huantai, 1.723 µg (0.224–49.772 µg) in Huaian, and 2.685 µg (1.006–14.534 µg) in Fusui. The averaged FB1 daily dietary intake was 92.4 µg (range = 55.0–362.1 µg) for residents of Huantai, 460.0 µg (83.2–2894.5 µg) in Huaian, and 138.6 µg (30.0–10,541.6 µg) in Fusui. These data suggest that the co-exposure to AFB1 and FB1 in residents of rural China may contribute to the aetiology of human chronic diseases in high-risk areas.

Silage is one of the most important feed sources for bovines. Mycotoxin contamination of feedstuffs is a worldwide concern. The aim of this study was to compare mycobiota and levels of aflatoxin B₁ (AFB₁), fumonisin B₁ (FB₁), deoxynivalenol (DON), zearalenone (ZEA) and patulin (PAT) in corn trench silos and silo bags. Dry matter was higher in trench silos. Counts varied from not detected to 10⁸ CFU g⁻¹ in both trench silos and silo bags. Isolation frequencies of Aspergillus spp. and Fusarium spp. were higher in trench silos, whereas Penicillium spp. was higher in silo bags. Silo bags showed less diversity than trench silos. Strains isolated produced AFB₁, FB₁ and PAT. In trench silos, AFB₁ was the only mycotoxin detected (1-160 µg kg⁻¹). In silo bags AFB₁ levels varied from 5.8 to 47.4 µg kg⁻¹. DON was detected in two silo bag samples. When handling is adequate the reduction of mould and mycotoxin contamination in silo bags is considerable. This study will enable estimation of the mycotoxicological risk of different ensiling practices and determination of the most adequate method to minimize economic losses and reduce hazard to animal and human health.