Fields Institute Communications

This study evaluated aqueous Vernonia amygdalina leaf extract in drinking water as a mitigation strategy against Aflatoxin B1-induced toxicity in broilers, focusing on performance, haematology, serum biochemistry, pro-inflammatory cytokines, cellular stress markers, and liver histology. Two hundred and forty (240) day-old chicks (mixed sex), of the Cobb 500 breed were divided into four groups: control (CONT), AFB1-exposed (AFLB1), and two treatment groups (VE1AF and VE2AF) receiving 0.5 mg/kg AFB1 and Vernonia amygdalina aqueous extract at 1 g/L and 2 g/L, respectively. At 42 days, VE1AF and VE2AF chickens showed higher (P < 0.05) final weights and weight gains than CONT and AFLB1 groups. The red blood cells, packed cell volume, haemoglobin, and white blood cell counts were higher (P < 0.05) in CONT, VE1AF, and VE2AF groups compared to AFLB1. Mean cell volume, and mean cell haemaoglobin were higher (P < 0.05) in AFLB1 and VE2AF. Serum analysis revealed lower (P < 0.05) total protein, globulin, and albumin in AFLB1, which were restored by the extract. The tumor necrosis factor-α, interleukin-6, interleukin-1β, and interferon-γ, were elevated (P < 0.05) in AFLB1 but reduced in VE1AF and VE2AF. The heat shock protein 70, 8-hydroxy-2'-deoxyguanosine and adiponectin levels were higher (P < 0.05) in AFLB1, but were normalized by the extract in VE1AF and VE2AF. Leptin and triiodothyronine levels were significantly (P < 0.05) better in VE1AF and VE2AF, compared to AFLB1. Liver histology showed reduced inflammation in VE1AF and VE2AF, with near-normal hepatic architecture. In conclusion, Vernonia amygdalina leaf extract effectively counteracts AFB1 toxicity, enhancing overall health and performance in broiler chickens.

Abstract Aflatoxins are potent carcinogens and pose significant risks to food safety and public health worldwide. Aflatoxins include Aflatoxin B1 (AFB1), Aflatoxin B2 (AFB2), Aflatoxin G1 (AFG1), Aflatoxin G2 (AFG2), and Aflatoxin M1 (AFM1). AFB1 is particularly notorious for its carcinogenicity, classified as a Group 1 human carcinogen by the International Agency for Research on Cancer (IARC). Chronic exposure to aflatoxins through contaminated food and feed can lead to liver cancer, immunosuppression, growth impairment, and other systemic health issues. Efforts to mitigate aflatoxin contamination have traditionally relied on chemical treatments, physical separation methods, and biological degradation. However, these approaches often pose challenges related to safety, efficacy, and impact on food quality. Recently, cold plasma treatment has emerged as a promising alternative. Cold plasma generates reactive oxygen species, which effectively degrade aflatoxins on food surfaces without compromising nutritional integrity or safety. This review consolidates current research and advancements in aflatoxin detoxification, highlighting the potential of cold plasma technology to revolutionize food safety practices. By exploring the mechanisms of aflatoxin toxicity, evaluating existing detoxification methods, and discussing the principles and applications of cold plasma treatment. Graphical Abstract

Abstract Aflatoxin contamination of corn can occur when developing kernels are infected by the plant pathogen Aspergillus flavus. One route of infection is from airborne conidia. We executed a series of experiments within the corn canopy during two growing seasons and in two states to document the abundance and dynamics of the airborne A. flavus population. We did not observe any significant diurnal changes in the conidial density (p = 0.171) or any effect of sampler height (p = 0.882) within the canopy. Significant changes (p < 0.001) were noted during the season, with a trend towards increased airborne populations with later stages of corn development and more than a 20-fold increase from July to August. The median aflatoxigenicity of airborne isolates from a corn canopy in Texas was about 50 times higher than the corresponding population in Mississippi. It was also noteworthy that highly aflatoxigenic, weakly sporulating S-morphotypes accounted for 14–30% of the airborne isolates in Mississippi at a site with historically rare abundance of S-morphotypes. The genetic diversity was high among the 140 analyzed airborne isolates, with 76 unique haplotypes identified and 55 haplotypes occurring only in 1 isolate. Even in the context of this highly diverse population, a haplotype matching that of a commercial biocontrol strain was found in 13 of the 70 isolates from Mississippi and 1 of the 70 isolates from Texas. The airborne A. flavus population is genetically diverse (Shannon’s index = 1.4 to 1.6), similar to grain samples in other surveys, and much less aflatoxigenic in Mississippi than in Texas.

Abstract Mycotoxin exposure from contaminated food is a significant global health issue, particularly among vulnerable children. Given limited data on mycotoxin exposure among Namibian children, this study investigated mycotoxin types and levels in foods, evaluated dietary mycotoxin exposure from processed cereal foods in children under age five from rural households in Oshana region, Namibia. Mycotoxins in cereal-based food samples (n = 162) (mahangu flour (n = 35), sorghum flour (n = 13), mahangu thin/thick porridge (n = 54), oshikundu (n = 56), and omungome (n = 4)) were determined by liquid chromatography-tandem mass spectrometry. Aflatoxin B1 (AFB1, 35.8%), zearalenone (27.2%), fumonisin B1 (FB1, 24.1%), citrinin (CIT, 12.4%) and deoxynivalenol (10.5%) were the major mycotoxins quantified. Food samples (35.8% (n = 58) and 6.2% (n = 10)) exceeded the 0.1 µg/kg AFB1 and 200 µg/kg FB1 EU limit for children’s food, respectively. Several emerging mycotoxins including the neurotoxic 3-nitropropionic acid, moniliformin (MON), and tenuazonic acid were quantified in over 50% of all samples. Co-occurrence of AFB1, CIT, and FB1 detected in 4.9% (n = 8) samples, which could heighten food safety concerns. Regarding exposure assessment and risk characterization, average probable dietary intake for AFB1 from all ready-to-eat-foods was 0.036 µg/kg bw/day, which resulted in margin of exposures (MOE) of 11 and 0.65 risk cancer cases/year/100,000 people, indicating a risk of chronic aflatoxicosis. High tolerable daily intake values for FB1, and MOE for beauvericin and MON exceeded reference values. Consumption of a diversified diet and interventions including timely planting and harvesting, best grain storage, and other standard postharvest food handling practices are needed to mitigate mycotoxin exposure through contaminated cereal foods and to safeguard the health of the rural children in Namibia.

Mycotoxins are secondary metabolites of fungi and represent a serious problem for human health. Due to growing interest, various aspects have been widely studied by scientific groups. One of these aspects relates to the food industry and associated beer production. Mycotoxins can be present in the basic raw materials for beer production as well as in brewed beer. Problematic mycotoxins that pose a serious risk of toxicity are aflatoxins especially aflatoxin B1 (AFB1), fumonisins (FBs), and zearalenone (ZEN) and its metabolites, deoxynivalenol (DON) including its acetylated forms and also the modified form deoxynivalenol-3-glucoside (DON-3G), T-2 toxin, HT-2 toxin, and ochratoxin A. The Research Institute of Brewing and Malting has been dealing with the issue of mycotoxins since 2008. This study describes the analysis of the above mycotoxins during 2020–2024 in barley (n = 775), malt (n = 751), and commercially available beers (n = 522) using QuEChERS, immunoaffinity columns, and UPLC-MS/MS. The results showed positive samples of mycotoxins in brewing and malting matrices at the level of micrograms per kilogram (barley, malt) and nanograms per liter for beer. Therefore, it is a residual concentration and the accurate quantitative determination of mycotoxins, correct interpretation of the results in connection with toxicological values, and the maximum permissible levels of mycotoxins play a key role in global food safety and consumer protection.

Abstract Molds of the genus Fusarium infect nearly all types of grain, causing significant yield and quality losses. Many species of this genus produce mycotoxins, which pose significant risks to human and animal health. In beer production, the complex interaction between primary fungal metabolites and secondarily modified mycotoxins in barley, malt, and beer complicates the situation, highlighting the need for effective analytical methods to quickly and accurately monitor these toxins. We developed and validated a liquid chromatography-tandem mass spectrometry (LC–MS/MS) method to simultaneously analyze 14 Fusarium toxins, including modified forms (deoxynivalenol (DON), DON-3-glucoside, 3-acetyl-DON, 15-acetyl-DON, nivalenol, fusarenone X, HT-2 toxin, T-2 toxin, the enniatins A, A1, B, B1, beauvericin, and zearalenone) in barley and throughout the malting process. Stable isotope dilution assays (SIDAs) and matrix-matched calibration were used for quantification. A micro-malting setup was established to produce Fusarium-contaminated barley malt under reproducible conditions using targeted inoculation with F. culmorum. Mycotoxins were quantified throughout the malting process and compared to the content of fungal DNA. Further, the impact of various malting parameters was investigated, thus revealing that different malting scenarios exhibited different toxin enrichment patterns. We demonstrated that mycotoxin concentration and the ratio of DON to DON-3-glucoside changed throughout the malting processes, depending on fungal spore concentrations, germination temperature, and malting temperature. The study highlights the complexity of mycotoxin dynamics in malt production and the importance of optimized processing conditions to minimize toxin levels in final malt products.

Aflatoxins (AF), fungal metabolites, can contaminate feed in favorable environments, posing health risks to humans and animals. Dairy cows exposed to aflatoxin B1 (AFB1) excrete its metabolite, aflatoxin M1 (AFM1), in milk, compromising its safety. The current study examined the use of an AF binder in dairy feed concentrates on farms in Rwanda to mitigate AFM1 in milk. It was conducted in Nyagatare district, peri-urban areas of Kigali (Center), and Huye district (South) with 42 farmers randomly selected from a previous study evenly divided into control and intervention groups. The intervention group received an AF binder (Novasil™ Plus) and training on both the usage of the binder and AF prevention in dairy feed. Sociodemographic and dairy management data were collected through semi-structured questionnaires. Farmers in both the treatment and control groups were visited at regular intervals over a 3-month time period (five total visits per farmer) to collect samples of milk and feed for the quantification of AFM1 and AFB1, respectively, and to interview participants. The use of the AF binder evidenced a significant reduction in milk AFM1 contamination between the intervention (mean, 0.15 µg/l; median, 0.13 µg/l) and control groups (mean, 0.30 µg/l; median, 0.24 µg/l) (p < 0.05). AFB1 was detected in the feed concentrate at a mean concentration of 32.2 µg/kg (median, 36.2 µg/kg); however, mean AFM1 levels in both groups (i.e., control and intervention) did not exceed the Rwanda Standards Board (RSB) regulatory limit of 0.5 µg/kg AFM1. AF binders show potential as a low-cost strategy to reduce AFM1 contamination in the Rwandan context. However, there is a need for clear standards on the registration and use of binders before they are deployed in-country and to claim their mitigating effects on AFM1 in milk.

The silymarin complex extracted from milk thistle provides significant health benefits, particularly due to its antioxidant and hepatoprotective properties. However, plant substances can be contaminated by a number of fungi types and their secondary metabolites—mycotoxins. This work deals with the determination of aflatoxins and zearalenone and its metabolites in 39 different samples grown in 2020 and 2021. Analysis of mycotoxins was performed by UHPLC-MS/MS after immunoaffinity column AFLAPREP® and EASI-EXTRACT® ZEARALENONE clean-up. The presence of aflatoxins was not confirmed in the monitored samples, but 1/3 of the samples were contaminated with zearalenone in the range of 2.8–378.9 µg/kg. Metabolites of zearalenone such as α-zearalenol, α-zearalanol, and β-zearalanol were not detected in any of the samples. β-Zearalenol was found in two samples (2.6 µg/kg and 29.8 µg/kg).

The infant flours produced in Burkina Faso are essentially a mixture of cereals and legumes. These raw materials are frequently contaminated with mycotoxins which pose a huge food safety and public health threat. The objective of this study was to determine mycotoxin levels in raw materials and infant flours in Ouagadougou and to investigate the impact of decontamination on the raw materials used in infant flour production. A total of 22 cereals and 17 legumes as raw materials and 26 infant flour samples were analysed for aflatoxins, fumonisin B1 (FB1), and ochratoxin A (OTA) by liquid chromatography coupled to tandem mass spectrometry, while saline treatment and hand-sorting of grains in mycotoxin reduction were tested. All the samples of raw materials and infant flours were contaminated with aflatoxins, whereas 20.5% and 38.5% of raw materials and 57.7% and 61.5% of infant flours, respectively, were contaminated by FB1 and OTA. These decontamination assays significantly reduced the levels of mycotoxins. AFB1 was reduced by 48% after soaking of maize for 6 h in a 6% NaCl solution. Sorting resulted in a 92% reduction in AFB1 content in peanut. However, soaking in saline solution did not reduce the FB1 and OTA contents. Sorting did not also reduce FB1 contents in peanut. Sorting and soaking in 6% saline solution for 6 h are production processes that lead to a reduction in the level of contamination by aflatoxins in maize and peanut used as raw materials for infant flour production.

Aflatoxins are toxic secondary metabolites produced by Aspergillus species that infect staple foods like maize causing threat to public health and economic impacts. The use of atoxigenic Aspergillus species is considered one of the promising technologies to prevent aflatoxin contamination in maize. Tanzania approved the use of aflatoxin biocontrol (Aflasafe®) in 2018 and introduced it to eight districts. Adoption and effectiveness of this technology depend on many factors including application of pre- and post-harvest practices. There is scant information on awareness of biocontrol and factors which influence the adoption and effectiveness of this technology. A cross-sectional study was conducted in Tanzania to assess awareness and identify factors influencing adoption of the technology. Data was collected from 334 smallholder farmers in Kiteto and Chemba districts and analyzed using SPSS version 20; p-values < 0.05 using a two-tailed test were considered statistically significant. Results indicated 95.4% are not aware and that only 2.7% of the farmers had used biocontrol technology. The use of biocontrol was significantly associated with high income level (p = 0.001) and exposure to print media (p = 0.03) and radio (p = 0.008). The use of hybrid seed (p = 0.01), grazing (p = 0.017), and rotation of crops on yearly basis (p = 0.024) were also significantly associated with the use of biocontrol. Income limits the use of biocontrol, requiring government subsidies for Aflasafe and a premium market for aflatoxin-free maize. Aflatoxin awareness and sensitization on adherence to good pre-harvest practices should be emphasized to enhance adoption of the technology.

Abstract Mycotoxins such as aflatoxins (AFs), fumonisins (FBs), zearalenone (ZEN), and deoxynivalenol (DON) pose a risk to public health due to their carcinogenic potency (AFs and FBs) and anti-nutritional effects. The hazards associated with mycotoxins are accentuated where food management practices, control, and regulatory systems from farm to plate are sub-optimal. Information on the frequency of these mycotoxins in rice commercialized in markets in sub-Sahara Africa (SSA) is limited. The current study examined AF concentrations in 527 rice samples collected from 54 markets in five SSA countries. Grain quality characteristics, processing methods, and origin of samples were contrasted with toxin levels. In total, 72% of the samples had detectable AFs levels (range = 3.0 to 89.8 µg/kg). Forty-seven percent (47%) of the samples had AFs above 4 µg/kg, the European Union maximum level (ML), and were evaluated for cooccurrence with FBs, ZEN, and DON. Total AFs and ZEN cooccurred in 40% of the samples, and 30% of the positive ZEN samples had concentrations above the ML of 75 µg/kg. Total AFs did not co-occur with FBs and DON. Multivariate analysis revealed that length-to-width ratio (p < 0.0001), mixed variety for width (p = 0.04), and chalkiness (p = 0.009) significantly influenced aflatoxin concentrations. Slender grains had higher AFs concentrations than bold and medium grains (p < 0.0001). Possible strategies to mitigate mycotoxin contamination in rice include improving grain quality traits and practicing proper drying and hermetic storage before and after milling. These findings provide valuable insights for both domestic and international actors in establishing and strengthening regulations and management systems to mitigate rice mycotoxin contamination.

Aflatoxin B1 (AFB1) and fumonisin B1 (FB1) are poisons that contaminate poorly stored staple foods in resource-limited settings. Antenatal AFB1 and FB1 exposure may cause anaemia. We aimed to determine the associations of urinary aflatoxin M1 (AFM1) and FB1, biomarkers of AFB1 and FB1 exposure, respectively, with erythrocyte parameters and anaemia. A retrospective cross-sectional study was conducted in 68 HIV-infected and 61 HIV-uninfected pregnant women ≥ 20 weeks gestational age in Harare, Zimbabwe. AFM1 and FB1 were measured in urine via competitive ELISA, and levels were grouped into tertiles. The erythrocyte parameters assessed were haemoglobin (Hb), mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red blood cell (RBC), haematocrit (HCT), and red blood cell distribution width. Associations of urinary AFM1 and FB1 with erythrocyte parameters, and anaemia were assessed in a multiple regression controlled for potential confounders. The presence of FB1 in urine decreased Hb levels in all women (β= −0.98, 95% CI: −1.94, 0.02) and HIV-uninfected (β= −1.99, 95% CI: −3.71, −0.26). FB1 tertile 3 decreased Hb levels (β= −0.88, 95% CI: −1.74, 0.01) and HCT levels (β= −2.65, 95% CI: −5.26, 0.03) in HIV-infected. AFM1 tertile 2 decreased RBC levels in HIV-infected (β= −0.34, 95% CI: −0.71, −0.03). The presence of FB1 in urine increased anaemia risk in HIV-uninfected (OR: 10.68 95% CI: 1.02, 112.34). AFM1 tertile 2 increased macrocytic anaemia risk in HIV-infected (OR: 13.72, 95% CI: 0.92, 203.55). There is need to ensure food safety through monitoring and nutritional interventions to improve maternal-infant health outcomes. 

Abstract Mycotoxins—secondary metabolites produced by filamentous fungal species—occur as a global problem in agriculture due to the reduction in crop quality and the negative effects on human and animal health. There is a need to develop environment-friendly methods of detoxification. In recent years, a number of biological methods for the removal/degradation of mycotoxins have been described. One of them—particularly interesting due to its high effectiveness—is mycoremediation, which involves the ability of Pleurotus spp. mushrooms to remove toxic contaminants from the environment and food. Pleurotus spp. biosynthesizes ligninolytic enzymes, such as laccase and manganese peroxidase that are the main factors of enzymatic degradation of various pollutants, including mycotoxins. The degradation process of mycotoxins (especially aflatoxins) with the participation of isolated enzymes reaches approximately 30–100%, depending on the culture conditions, substrate, and mediators used. In the food industry, their application may include, among others, the detoxification of animal feed from mycotoxins or fermentation products (e.g., juices and wines). While these applications are promising, they require further research to expand toxicological knowledge and optimize their use. This review presents current research on this new and very promising topic related to the use of edible Pleurotus spp. mushrooms in the process of biological degradation of toxic fungal metabolites.

To date, there are more than 80 ergot alkaloids identified; their distribution depends on different factors (e.g. geographic regions, host plants). These toxins can cause acute and chronic toxic effects on human health and commonly infect cereal crops such as triticale and rye, wheat, barley and oats. Considering the growing consumption of plant-based foods, the European Food Safety Authority has highlighted the need to develop risk assessment strategies. This work focused on the optimization of extraction efficiency, to quantify the main ergot alkaloids and their epimers, that are available on the market without any legal restriction (ergosine, ergocristine, ergocriptyne, ergocornine, ergosinine, ergocristinine, ergocriptinine and ergocorninine). Considering the quantification of 8 out of 12 regulated compounds by EU (sum of –ine and –inine forms), this approach can be defined as a screening method for a reliable estimation of the risk, specifically devoted to industrial stakeholders that can then possibly outsource to authorized external labs only the samples suspected of significant positivity. The effectiveness of three different extraction conditions (acidic, alkaline and neutral) followed by a rapid clean-up using dispersive solid-phase extraction with C18 sorbent was evaluated by ultra-performance liquid chromatography tandem quadrupole mass spectrometry (UPLC-MS/MS), resulting in a short chromatographic run (16 min). The method was developed and validated in five different cereal production chains (rye, oat, wheat, wheat gluten and baby food). The applicability of the method was examined by analyzing a set of 54 samples, including also other cereals like spelt, tritordeum and triticale, and evaluating also some reference materials.

Deoxynivalenol ( DON) is one of the most harmful mycotoxins in food or feed or Traditional Chinese Medicine. An efficient and applicable method for the detoxification of DON is urgently developed. 1152 strains were isolated from the intestinal contents of crucian. Morganella morganii YC12-C3 and Enterococcus faecalis YC12-C10 were screened with the highest degradation rate of DON via HPLC methods. The optimal degradation condition of YC12-C3 and YC12-C10 is co-cultured 24 h and 36 h at 28 ℃ in LB medium with pH 7 and 1.0% inoculation dosage, respectively. LC–MS/MS and 1H NMR results show that YC12-C10 and YC12-C3 can transform DON to 3-deoxy-6-demethanol-DON, a new metabolite biotransformed from DON, by deoxidization at C3 hydroxy and de-methanal reaction at methanol moiety of C6. In addition, the DON-degradation in agricultural material assay showed that YC12-C10 and YC12-C3 can degrade 150 μg·kg−1 DON in Coix lacryma-jobi, with a degradation rate of 68.89% and 59.94%, respectively. This result shows that YC12-C10 and YC12-C3 have a sound efficiency in removing DON ability in Coix lacryma-jobi, providing a new strain resource and application technique for biological detoxification of DON in food or feed or TCM industry.