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AbstractBackgroundObjectivesMethodsResultsConclusionsMycotoxins, fungal metabolites prevalent in many foods, are recognized for their role in carcinogenesis, especially when interacting with oncogenic viruses.This scoping review synthesizes current evidence on the human cancer risk associated with mycotoxin exposure and oncogenic virus infections.Searches were conducted on PubMed, Embase, and Web of Science. Studies were selected based on the PECOS framework. Data extraction involved narrative and qualitative presentation of findings, with meta-analysis where feasible. Risk of bias and outcome quality were assessed using the OHAT tool and GRADE approach.From 25 included studies, 18 focused on aflatoxins and hepatitis viruses in hepatocellular carcinoma (HCC). Four studies examined aflatoxin B1 (AFB1) and human papilloma virus (HPV) in cervical cancer, while three investigated AFB1 with Epstein-Barr virus (EBV) in lymphomagenesis. The review highlights a significant synergistic effect between AFB1 and hepatitis B and C viruses in HCC development. Significant interactions between AFB1 and HPV, as well as AFB1 and EBV, were observed, but further research is needed.The synergistic impact of mycotoxins and oncogenic viruses is a critical public health concern. Future research, especially prospective cohort studies and investigations into molecular mechanisms, is essential to address this complex issue. [ABSTRACT FROM AUTHOR]

A method based on a hazard analysis critical control point (HACCP)-based standard operating procedure (SOP) has been developed and implemented in community-based (CB) production of complementary foods (CFs) in agrarian regions in rural Ethiopia. The objective of this study was to validate the method in another setup, more specifically in cash crop (coffee and khat) producing communities in Southern Ethiopia. A quasi-experimental study was employed for the method validation. Two villages in the Gedeo zone, Southern Ethiopia were selected and a total of 40 mother-child pairs, 20 in each village, were involved in the study. Sociodemographic data, knowledge, and complementary feeding practices were collected using pretested questionnaires. The training was given to each mother for 1 h about community-based CFs production and implementation of the method. Then, 30 CF samples were collected and the level of aflatoxins (AFB1, AFB2, AFG1, and AFG2) was determined using a validated method. The majority (77.5%) of the mothers understand the importance of complementary feeding and give it to their infants beyond 6 months. Nearly two-thirds (62.5%) of the mothers had knowledge about the health impacts of aflatoxins, but the remaining 37.5% lacked awareness about the health risks of aflatoxins. Mothers were very well-motivated for the training and they promised to implement the knowledge and skill gained from the training for improving the nutritional status of their children. Overall, 23.3% of the CFs were contaminated by aflatoxins. AFB1 and AFG1 were detected in 6.7 and 16.7% of the CF samples, respectively. In cash crop-producing communities around the country, the HACCP-based SOP is easily validated in CB CFs production. Therefore, a scaleup of the method at the national level and beyond is recommended. [ABSTRACT FROM AUTHOR]

Mycotoxins, being a threat to animal and human health, contribute significantly towards economic losses in the poultry sector. A liquid chromatography-mass spectrometry-based study was conducted on poultry feed samples collected from Punjab, Pakistan to evaluate the prevalence, contamination levels, and co-occurrence of multi-mycotoxins across different processed forms of the feed, types of utilities and sampling regions. All samples were found to be contaminated with aflatoxin B1 (AFB1) and fumonisin B1 (FB1). The European Commission (EC) maximum level for AFB1 in complete feedingstuffs in poultry and guidance values for FB1 and zearalenone (ZEN) were exceeded in 73%, 2%, and 14% of the contaminated samples, respectively. The corresponding median values were 39.9 µg/kg, 205 µg/kg, and 34.5 µg/kg. In addition to exceeding contamination levels, a varying co-occurrence of three to fourteen mycotoxins was observed in each of the feed samples that calls for mitigation measures to safeguard the feed and its ingredients. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is a type B trichothecene mycotoxin with worldwide high incidence in feed which is produced by Fusarium species. Strategies are needed to eliminate its health risk for livestock and to minimise its economic impact on production. In order to assess the efficacy of potential physical, chemical and biological DON detoxifying agents, a good in vitro model is necessary to perform a fast and high-throughput screening of new compounds before in vivo trials are set up. In this paper, an in vitro model was developed to screen potential commercial products for DON degradation and detoxification. Contaminated feed with potential detoxifying agents are first applied to a simulated gastrointestinal tract (GIT) of a pig, after which detoxification is assessed through a robust, inexpensive and readily applicable Lemna minor L. aquatic plant bioassay which enables evaluation of the residual toxicity of possible metabolites formed by DON detoxifying agents. The GIT simulation enables taking matrix and incubation parameters into account as they can affect the binding, removal or degradation of DON. One product could reduce DON in feed in the GIT model for almost 100% after 6 h. DON metabolites were tentatively identified with LC-MS/MS. This GIT simulation coupled to a detoxification bioassay is a valuable model for in vitro screening and assessing compounds for DON detoxification, and could be expanded towards other mycotoxins. [ABSTRACT FROM AUTHOR]

The aim of this study was to systematically review associations between dietary mycotoxins exposure and child growth and morbidity of children aged 5 years or younger. Peer-reviewed literature was searched in MEDLINE, EMBASE, COCHRANE, CINAHL, Web of Science, and PsycINFO. Experimental and observational studies were considered. The exposures were dietary mycotoxins during pregnancy, lactation and childhood, and mycotoxins concentrations in the diet, breast milk, urine, and blood. From a total of 4869 references, 86 full-text papers were extracted of which 50 were included in this review. The methodological quality and risk of bias were evaluated and quality of the collective evidence was assessed using GRADE. Uncertainty remains whether mycotoxins exposure affects child growth, immunity and mortality and the overall quality of the evidence is very low. Overall however, we cannot rule out a possible association between dietary mycotoxins, in particular, AF and FUM and child malnutrition. Our analyses were limited by the reporting quality, difference in findings, heterogeneity of outcomes, mycotoxins detection methods, and the observational nature of most studies. Robust study designs with adequate sample size, use of validated biomarkers of exposure and assessment of co-occurrence of mycotoxins and their synergistic effects are required to provide the further evidence regarding a potential effect of dietary mycotoxins exposure on child growth and immunity. [ABSTRACT FROM AUTHOR]

Mycotoxins are unavoidable environmental contaminants, which are found throughout the food chain, particularly in cereals. Mycotoxin management is not effective in developing countries, such as Zimbabwe, due to resource constraints, yet human health risk is evident. Various practical mitigation strategies that can be employed to decrease human dietary exposure to mycotoxins as a means of preliminary steps towards risk management are discussed. These strategies were stratified into two categories. First, crop/commodity-centred strategies, mainly the pre-harvest actions of cultivar selection, bio-control, as well as good agricultural practices (GAP), and the post-harvest actions including timeous harvesting, appropriate drying and storage technologies, are elaborated making use of hazard analysis critical control points (HACCP) principles. The role of legislation is also explored as a crop/commodity centred mitigation strategy. Second, human-centred strategies anchored on dietary diversity and the use of socio-cultural approaches as a direct means of reducing mycotoxin exposure are discussed. Finally, an integrated science-based mycotoxin management strategy, encompassing targeted legislation on mycotoxins, consumer education and information sharing, human and institutional capacity building, training and financing, is suggested in addition to GAP, as a means of reducing human health risk associated with mycotoxin exposure in Zimbabwe.HighlightsFarm-to-fork HACCP-based mycotoxin managementHuman-centred mycotoxin management approaches are keyAgronomy, technology and legislation critical in reducing mycotoxin exposure. [ABSTRACT FROM AUTHOR]

This research aimed at evaluating the safety, and the type, level and prevalence of mycotoxins in grain sorghum of four sub-Saharan African (SSA) countries (Burkina Faso, Ethiopia, Mali and Sudan). A multi-analyte LC-MS/MS method for quantification of 23 mycotoxins (nivalenol, deoxynivalenol, fusarenon X, neosolaniol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, diacetoxyscirpenol, roquefortine C, HT-2 toxin, alternariol, T-2 toxin, FB1, FB2, FB3, zearalenone, aflatoxin G1, aflatoxin G2, aflatoxin B1, aflatoxin B2, sterigmatocystin, OTA, altenuene, alternariol monomethylether) was applied to different sorghum matrices. Of the 1533 analysed samples, 33% were contaminated with at least one of the following mycotoxins: aflatoxins, fumonisins, sterigmatocystin, Alternaria toxins, OTA and zearalenone. Country of origin, colour, source and collection period of sorghum samples significantly influenced the type, level and prevalence of mycotoxins. Sterigmatocystin (15%), fumonisins (17%) and aflatoxins (13%) were the most prevalent. FB1 (274 ± 585 µg/kg) had the highest mean concentration followed by FB2 (214 ± 308 µg/kg) while diacetoxyscirpenol (8.12 ± 19.2 µg/kg) and HT-2 (11.9 ± 0.00 µg/kg) had the lowest concentrations. Neosolaniol, fusarenon-X, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, T-2 toxin, nivalenol and roquefortine C were not detected in any of the samples. Sudan had the lowest prevalence and mean concentration of all mycotoxins. Pink sorghum had the highest concentrations of fumonisins and aflatoxins. Mycotoxins from Aspergillus spp. and Alternaria spp. are the mycotoxins of concern in SSA grain sorghum with regard to prevalence, concentration and possible health risk from exposure. Based on the performed risk characterisation, daily consumption of sorghum containing aflatoxins, alternariol, alternariol monomethyl ether, sterigmatocystin and OTA could result in exceeding the established health-based guidance values for these toxins. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the occurrence and levels of total aflatoxins in complementary foods (CFs) and their ingredients. A total of 126 samples collected from 20 Districts from Amhara, Tigray, Oromia, and Southern Nations Nationalities and Peoples (SNNP) regions were analysed for levels of total aflatoxins using enzyme linked immunosorbent assay (ELISA). Aflatoxins were detected in 62 out of 66 pre-milling samples with mean range of 0.3-9.9 µg/kg. Aflatoxins were also detected in 19 out of 20 post-production CFs and in all of the one-month stored CFs at households and grain banks, with a mean range of 0.5-8.0, 3.6-11.3, and 0.2-12.4 µg/kg, respectively. Overall, 3 out of 126 samples exceeded the maximum limit (10 µg/kg). Although most aflatoxin levels were below the maximum limit and thus considered to be safe for consumption, more effort should be implemented to reduce contamination, as these CFs are intended for consumption by young children. [ABSTRACT FROM AUTHOR]

The direct measurement of biomarkers of exposure in biological fluids such as urine has become important for assessing aflatoxin exposure in humans as it is the only tool that integrates exposures from various routes. For this reason, a study was conducted to assess aflatoxin exposure among young children in Ethiopia using urinary biomarkers. A cross-sectional study was conducted in ten Woredas (Districts) from Amhara and Tigray regional states of Ethiopia including 200 children (aged 1–4 years). A total of 200 urine samples were collected from 200 children and assessed for the levels of aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2) and aflatoxin M1 (AFM1) using a validated LC-MS/MS method. Aflatoxins were detected in 34/200 (17%) of the urine samples whereby four out of five analysed aflatoxins were detected. AFM1 was detected in 14/200 (7%) of the urine samples in a range of 0.06–0.07 ng/mL. AFB2, AFG2 and AFG1 were detected in respectively 9/200 (4.5%), 6/200 (3%) and 5/200 (2.5%) of the urine samples whereas AFB1 was not detected in any of the samples. In this study, there was no association between the different malnutrition categories (stunted, wasting and underweight) and aflatoxin exposure. However, the biomarker analysis showed a clear exposure of young children to aflatoxins. Therefore, awareness to the public is important to prevent potential health consequences of aflatoxins. [ABSTRACT FROM AUTHOR]

Following the discovery of aflatoxins in the early 1960s, there have been many studies leading to the uncovering of many mycotoxins and the understanding of associated health effects in animals and humans. Consequently, there has been a global increase in the number of countries with mycotoxin regulations in foods. However, many African countries have only regulations for aflatoxins (or a few other mycotoxins) in specific foods, or no regulations at all. This paper critically reviews the challenges thwarting the establishment of mycotoxin regulations and their impacts on human dietary mycotoxin exposure in Africa. Mycotoxin regulatory limits for different countries are compared with mycotoxin tolerable daily intakes established by international food safety bodies taking into account consumption patterns. The agrarian setup, food insecurity, and mycotoxin analytical challenges in African countries are discussed; and more feasible mycotoxin dietary exposure reduction strategies are proposed. [ABSTRACT FROM AUTHOR]

The fungusAspergillus flavusis known for its ability to produce the toxic and carcinogenic aflatoxins in food and feed. While aflatoxins are of most concern,A. flavusis predicted to be capable of producing many more metabolites based on a study of its complete genome sequence. Some of these metabolites could be of great importance in food and feed safety. Here we describe an analytical methodology based on Orbitrap HRMS technology that allows the untargeted determination of fungal metabolites, in support of the study of the function of genes involved in secondary metabolism in fungi. The applied strategy implies the detection and identification of differentially expressed metabolites in extracts of wild-type and mutant fungal strains, using Orbitrap high-resolution mass spectrometry (HRMS) accurate mass data. The suitability of this approach was demonstrated by the confirmation of previously characterised genes involved in the aflatoxin biosynthetic pathway, namely a polyketide synthase (pksA), an oxidoreductase (ordA) and a methyltransferase (omtA) gene. Subsequently, the proposed methodology was applied for the detection and identification of metabolites produced by a yet uncharacterised gene cluster inA. favus, cluster 23. Comparative Orbitrap HRMS analysis of extracts ofA. flavuswild-type strain and an over-expression mutant for the transcription factor of gene cluster 23 (lepE) demonstrated that this gene cluster is responsible for the production a set of 2-pyridone derivatives, the leporins. Besides the known derivatives leporin B and leporin B precursor that could be identified by automatic de-replication of the accurate mass data, five other compounds belonging to this class of fungal secondary metabolites were detected and identified for the first time, combining MS and multiple-stage MS data. [ABSTRACT FROM PUBLISHER]

Maize is one of the major staple foods of Sub-Saharan Africa and is consumed as whole or dehulled grain. In this region, where the environmental conditions favour fungal growth and mycotoxin production, the majority of the population are subsistence consumers who, unfortunately, have little or no access to mycotoxin testing of their food. In an attempt to develop feasible reduction strategies in dietary mycotoxin exposure of the population, a three-factorial design experiment was conducted to examine and compare the efficacy of hand sorting, flotation, dehulling and combinations thereof in removing naturally occurring aflatoxins, fumonisins, nivalenol, deoxynivalenol and alternariol in shelled white maize. Regression analysis was used to determine the significant (p < 0.05) process variables on the removal of mycotoxins from the maize. Results from this experiment indicated that hand sorting had the greatest effect on mycotoxin removal, while flotation yielded the least effect. In particular hand sorting left < 6% of aflatoxin B1 and < 5% of fumonisin B1. Based on these results, hand sorting of maize grains is being recommended as a last line of defence against mycotoxin exposure among subsistence consumers. [ABSTRACT FROM AUTHOR]

An immunogen synthesis strategy was designed to develop anti-deoxynivalenol (DON) monoclonal antibodies with low cross-reactivity against structurally similar trichothecenes. A total of eight different DON immunogens were synthesised, differing in the type and position of the linker on the DON molecule. After immunisation, antisera from mice immunised with different DON immunogens were checked for the presence of relevant antibodies. Then, both homologous and heterologous enzyme-linked immunosorbent assays (ELISAs) were performed for hybridoma screening. Finally, three monoclonal antibodies against DON and its analogues were generated. In addition, monoclonal antibody 13H1 could recognise DON and its analogues in the order of HT-2 toxin > 15-acetyldeoxynivalenol (15-ADON) > DON, with IC50ranging from 1.14 to 2.13 µg ml–1. Another monoclonal antibody 10H10 manifested relatively close sensitivities to DON, 3-acetyldeoxynivalenol (3-ADON) and 15-ADON, with IC50values of 22, 15 and 34 ng ml–1, respectively. Using an indirect ELISA format decreases the 10H10 sensitivity to 15-ADON with 92%. A third monoclonal antibody 2A9 showed to be very specific and sensitive to 3-ADON, with IC50of 0.38 ng ml–1. Using both 2A9 and 10H10 monoclonal antibodies allows determining sole DON contamination. [ABSTRACT FROM AUTHOR]

Mycotoxins are harmful food contaminants. Currently, human exposure assessment to these toxins is often based on calculations combining mycotoxin occurrence data in food with population data on food consumption. Because of limitations inherent to that approach, biomarkers have been proposed as a suitable alternative whereby a more accurate assessment of exposure at the individual level can be performed. The BIOMYCO study is designed to assess human mycotoxin exposure using urinary biomarkers of exposure. Over the different seasons of 2013 and 2014, morning urine is gathered in a representative part of the Belgian population according to a designed study protocol, whereby 140 children (3–12 years old) and 278 adults (19–65 years old) are selected based on random cluster sampling stratified for sex, age and geographical areas. Every participant completes a food frequency questionnaire to assess the consumption of relevant foodstuffs (n = 43) of both the day before the urine collection and the previous month. Validated multi-toxin LC-MS/MS methods are used to analyse aflatoxins, fumonisins, ochratoxin A, trichothecenes, zearalenone and their metabolites in morning urine. The study protocol is approved by the ethical committee of the Ghent University Hospital. Within this paper, study design and methods are described. The BIOMYCO study is the first study whereby a multi-toxin approach is applied for mycotoxin exposure assessment in adults and children on a large scale. Moreover, it is the first study that will describe the exposure to an elaborated set of mycotoxins in the Belgian population. In first instance, descriptive analysis will be performed, describing the exposure to mycotoxins for the child and adult group. Exposure of different subgroups will be compared. Furthermore, correlations between the mycotoxin concentrations measured and the food consumption reported will be estimated to explore whether the mycotoxin exposure could be explained by the consumption of certain foods. [ABSTRACT FROM AUTHOR]

The filamentous fungusAspergillus flavusis one of the most important species in theAspergillusgenus and is distributed worldwide as a prevalent aflatoxin-producing food and feed contaminant.A. flavuscontains more than 55 gene clusters that are predicted to encode proteins involved in secondary metabolite production. One of these, cluster 27, contains a polyketide synthase (pks27) gene that encodes a protein that is highly homologous to the aflatoxin cluster PKS. Comparative metabolomics, using ultra-high performance liquid chromatography (UHPLC) coupled to high resolution Orbitrap mass spectrometry (MS) was used to detect metabolites differentially expressed in theA. flavus wild-type and∆pks27 mutant strains. Metabolite profiling was aided by a statistical differential analysis of MS data using SIEVE software. This differential analysis combined with accurate mass data from the Orbitrap and ion trap multiple stage MS allowed four metabolites to be identified that were produced only by the wild-type culture. These included asparasone A (358 Da), an anthraquinone pigment, and related anthraquinones with masses of 316, 340 and 374 Da. These latter three compounds had similar fragmentation patterns to that of asparasone A. The 316 Da anthraquinone is particularly interesting because it is most likely formed by incorporation of seven malonyl-CoA units rather than the eight units required for the formation of asparasone A. The 340 and 374 Da metabolites are the dehydration and an oxy-derivative of asparasone A, respectively. Asparasone A was also identified in extracts from several otherAspergillusspecies. [ABSTRACT FROM AUTHOR]

In this work, molecularly imprinted polymers (MIPs) using the mycotoxin zearalenone (ZEA) as template were first synthesized via precipitation polymerization and then applied as specific adsorbents in molecularly imprinted solid-phase extraction (MISPE) cartridges. The MISPE procedure was optimized with 3 mL of acetonitrile for preconditioning, 1 mL of acetonitrile:H2O (60:40) for loading, 1 mL of acetonitrile:H2O (30:70), and 3 mL of methanol:acetic acid (95:5) for elution. The obtained MIPs showed high selectivity of 96.9% towards ZEA, and low cross-reactivity (1-20%) to other Fusarium mycotoxins including deoxynivalenol, nivalenol, HT-2 toxin and T-2 toxin. The cross-reactivity to fumonisin B1 amounted to 61%. The MISPE was applied for enrichment of ZEA, which was produced by Fusarium graminearum strains. An enrichment factor above 50 was reached. Recoveries of 1 µg/mL were between 90.8% and 99.6%. A small amount of ZEA was produced by 9 F. graminearum strains with a maximum of 13 µg, then purified by the developed MISPE and analyzed by LC-MS/MS. [ABSTRACT FROM AUTHOR]

A multi-analyte method for the liquid chromatography-tandem mass spectrometric determination of mycotoxins in food supplements is presented. The analytes included A and B trichothecenes (nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, neosolaniol, fusarenon-X, diacetoxyscirpenol, HT-2 toxin and T-2 toxin), aflatoxins (aflatoxin-B1, aflatoxin-B2, aflatoxin-G1 and aflatoxin-G2), Alternaria toxins (alternariol, alternariol methyl ether and altenuene), fumonisins (fumonisin-B1, fumonisin-B2 and fumonisin-B3), ochratoxin A, zearalenone, beauvericin and sterigmatocystin. Optimization of the simultaneous extraction of these toxins and the sample pretreatment procedure, as well as method validation were performed on maca (Lepidium meyenii) food supplements. The results indicated that the solvent mixture ethyl acetate/formic acid (95:5, v/v) was the best compromise for the extraction of the analytes from food supplements. Liquid-liquid partition with n-hexane was applied as partial clean-up step to remove excess of co-extracted non-polar components. Further clean-up was performed on Oasis HLB™ cartridges. Samples were analysed using an Acquity UPLC system coupled to a Micromass Quattro Micro triple quadrupole mass spectrometer equipped with an electrospray interface operated in the positive-ion mode. Limits of detection and quantification were in the range of 0.3-30 ng g-1 and 1-100 ng g-1, respectively. Recovery yields were above 60% for most of the analytes, except for nivalenol, sterigmatocystine and the fumonisins. The method showed good precision and trueness. Analysis of different food supplements such as soy (Glycine max) isoflavones, St John's wort (Hypericum perforatum), garlic (Allium sativum), Ginkgo biloba, and black radish (Raphanus niger) demonstrated the general applicability of the method. Due to different matrix effects observed in different food supplement samples, the standard addition approach was applied to perform correct quantitative analysis. In 56 out of 62 samples analysed, none of the 23 mycotoxins investigated was detected. Positive samples contained at least one of the toxins fumonisin-B1, fumonisin-B2, fumonisin-B3 and ochratoxin A. [ABSTRACT FROM AUTHOR]

A rapid antibody-based assay for the detection of ochratoxin A in cocoa powder is described, involving sequential clean-up and visual detection of the toxin ("clean-up tandem assay column"). The screening test was developed to have a cut-off level of 2 µg kg-1 and was shown to have false positive and false negative rates of 10 and 2%, respectively. Analysis of six samples can be carried out in the field in approximately 30 min by untrained workers. Using the proposed rapid screening test, 10 retail cocoa powders were found to contain no detectable levels of ochratoxin A (<2 µg kg-1). These samples were also found to be negative (<2 µg kg-1) when analysed using an LC-MS/MS method. [ABSTRACT FROM AUTHOR]