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Mycotoxins are fungal metabolites that contaminate crops, food, and animal feeds. Aflatoxins and fumonisins are among the mycotoxins that have been increasingly reported to affect health and productivity of livestock globally. Given that the health and productivity of livestock can directly influence human food safety and security, a study was conducted to assess the levels and factors for aflatoxin and fumonisin contamination in feed and feed ingredients in Rwanda. Aflatoxins and fumonisins were analyzed in 3328 feed and feed ingredient samples collected at six time points between March and October 2017 in all 30 districts of Rwanda. Of the 612 participants providing samples, there were 10 feed processors, 68 feed vendors, 225 dairy farmers, and 309 poultry farmers. Enzyme-Linked Immunosorbent Assay (ELISA) was used for aflatoxin and fumonisin analyses. Mean aflatoxin levels of 108.83 µg/kg (Median (MD): 43.65 µg/kg), 103.81µg/kg (MD: 48.4 µg/kg), 88.64 µg/kg (MD: 30.90 µg/kg), and 94.95 µg/kg (MD: 70.45 µg/kg) were determined for dairy farmers, poultry farmers, feed vendors, and feed processors, respectively. Mean fumonisin levels were 1.52 mg/kg (MD: 0.71 mg/kg), 1.21 mg/kg (MD: 0.56 mg/kg), 1.48 mg/kg (MD: 0.76 mg/kg), and 1.03 mg/kg (MD: 0.47 mg/kg) for dairy farmers, poultry farmers, feed vendors, and feed processors, respectively. Aflatoxin contamination was significantly affected by time of sampling and district from which feed samples originated ( p < 0.05). Fumonisins did not show any correlation trends. Ninety-two percent of survey participants were unaware of aflatoxins and fumonisins and their adverse effects. This study has provided the basic understanding of the extent of feed contamination across the country and has established a baseline for future interventions in Rwanda. Further studies are needed to explore strategies for mitigating mycotoxins in the feed value chain in Rwanda.

Maize (Zea mays), sorghum (Sorghum bicolor) and pearl millet (Pennisetum glaucum) are basic staple foods for many rural or poorer communities. These crops are susceptible to plant diseases caused by multiple species of Fusarium, some of which also produce mycotoxins, including fumonisins and moniliformin that are detrimental to both humans and domesticated animals. Eighteen potentially toxigenic Fusarium strains were isolated from maize (n = 10), sorghum (n = 7) and pearl millet (n = 1) growing in the same field in Nigeria. The 17 strains from maize and sorghum were all F. proliferatum and the one strain from pearl millet was F. pseudonygamai. Under conducive conditions, the 17 F. proliferatum strains produced fumonisins, 11 in relatively large quantities (700-17,000 mg total fumonisins, i.e., FB 1  + FB 2  + FB 3 /kg culture material), and six at <45 mg/kg. Ten F. proliferatum strains produced >100 mg of moniliformin per kg culture material with a maximum of 8900 mg/kg culture material. All strains could use all grains for growth and toxin production, regardless of the host from which they were isolated. Isolates varied in the amount of toxin produced on each substrate, with toxin production a property of the strain and not the host from which the strain was recovered. However, the extent to which a toxin-producing phenotype could be altered by the grain on which the fungus was grown is consistent with subtle genetic × environment interactions that require a larger data set than the one presented here to rigorously identify. In conclusion, there is significant variation in the ability of strains of F. proliferatum to produce fumonisins and moniliformin on maize, sorghum and millet. If the amount of toxin produced on the various grains in this study reflects real-world settings, e.g., poor storage, then the consumers of these contaminated grains could be exposed to mycotoxin levels that greatly exceed the tolerable daily intakes., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim of this study was to compare the fate of fumonisins in transgenic and non-transgenic corn during industrial dry milling. For this purpose, whole corn samples and their fractions (germ, pericarp, endosperm, corn meal, and grits) were collected from one of the major Brazilian milling plants, totaling 480 samples. There was no significant difference (p > 0.05) between mean fumonisin (FB 1 + FB 2 ) levels in transgenic (1130 μg/kg) and non-transgenic (920 μg/kg) whole corn. However, in non-transgenic germ, endosperm and corn meal fraction fumonisin levels were higher (2940 μg/kg, 250 μg/kg and 190 μg/kg, respectively) than in transgenic fractions (2180 μg/kg, 130 μg/kg and 85.0 μg/kg, respectively). Furthermore, the highest percentages of fumonisins were distributed in the germ, corresponding to about 87 and 76% of the total fumonisins present in the whole corn from non-transgenic and transgenic hybrids, respectively. Concerning the endosperm from non-transgenic and transgenic corn, approximately, 23% and 13% of the total fumonisins were retained after the dry milling. Further processing in corn meal (300 to 420 μm particle size) and grits (590 to 1190 μm) decreased the percentages of remaining fumonisins to 4% and 2% (transgenic) and 10% and 3% (non-transgenic corn), respectively. These results suggested that fumonisin concentration was higher in outer and inner non-transgenic fractions when compared to transgenic ones and that the fate of fumonisins during the industrial dry milling could be affected by the transgenic status. However, it was not possible to conclude that the difference was exclusively due to this variable.

Maize has become one of the most important crops for food and feed production-both as a silage and crop residue worldwide. The present study aimed to identify the co-occurrence of Fusarium subglutinans , Fusarium verticillioides , Trichoderma atroviride , Sarocladium zeae, and Lecanicillium lecanii on maize ear rot. Further, the accumulation of mycotoxins as secondary metabolites of Fusarium spp. in maize ear samples was also analyzed. Maize ear samples were collected between 2014 and 2017 from two main maize growing areas in Poland (Greater Poland and Silesia region). A significant difference was found in the frequency of two main Fusarium spp. that infect maize ears, namely F. subglutinans and F. verticillioides . In addition to Fusarium spp. T. atroviride , S. zeae, and L. lecanii were also identified. T. atroviride species was found in 14% of maize samples examined between 2014 and 2017, particularly with a high percentage of Trichoderma spp. recorded in 2014, i.e., in 31% of samples. However, mycotoxin content (beauvericin and fumonisins) varied, depending on both the location and year of sampling. The interaction of fungi and insects inhabiting maize ear and kernel is very complex and not yet elucidated. Therefore, further research is required in this area.

Fumonisins are mycotoxins (MTs) produced mainly by the fungus Fusarium verticillioides , the main pathogens of maize which cause ear rot. The aim of this work was to evaluate some factors that may lead to high fumonisin production by F. verticillioides in maize grains, correlating the pathogen inoculation method with different genotypes grown in four Brazilian states. Experiments were conducted in 2015-2016 in maize crops from experimental maize fields located in four distinct states of Brazil. Results showed that contamination by fumonisin mycotoxins occurred even on symptomatic or asymptomatic grains. In all municipalities, the samples showed levels of fumonisin B1 that were higher than would be tolerable for the human consumption of corn products (the current tolerance limit for fumonisin is 1.5 μg g -1 ). High severity of grains infected with F. verticillioides does not always show high concentrations of fumonisins. Environments with higher temperatures may influence the production of high concentrations of fumonisin in maize hybrids. Spray inoculation methods and inoculation at the center of spikes did not influence fumonisin concentrations. Results showed that the hybrids P3630H, P32R48 and P3250 presented higher disease severity, as well as higher mycotoxin levels in the studied locations with higher temperatures.

Due to the widely occurring co-contamination of mycotoxins in raw food materials, simultaneous monitoring of multiple mycotoxins is needed. Herein, we report the design and fabrication of an electrochemical immunosensor for simultaneous detection of two mycotoxins, fumonisin B1 (FB1) and deoxynivalenol (DON), in a single test. A dual-channel three-electrode electrochemical sensor pattern was etched on a transparent indium tin oxide (ITO)-coated glass via photolithography and was integrated with capillary-driven polydimethylsiloxane (PDMS) microfluidic channel. The two working electrodes were functionalized with gold nanoparticles and anti-FB1 and anti-DON antibodies. Tests were performed by incubating the working electrodes in a sample solution introduced in the PDMS channel. The formation of toxin-antibody immunocomplexes on the working electrode surface produced electrochemical signal responses, which were recorded and compared with control signal to quantify individual mycotoxin concentrations. Using this dual-channel ITO-microfluidic electrochemical immunosensor we achieved limits of detection (LODs) of 97 pg/mL and 35 pg/mL, respectively for FB1 and DON, and their corresponding linear ranges of detection were 0.3-140 ppb and 0.2-60 ppb. The sensor performance, which remained stable for two weeks under proper storage, was validated by testing with ground corn extract used as a real food matrix., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In Argentina, wheat is the most consumed cereal by the human population. Since fumonisins occurence in wheat grains and wheat-based products have been reported worldwide, a survey was conducted in order to determine fumonisin contamination in 91 wheat-based products (white wheat flour samples, wheat flour used at bakery products and whole-wheat flour samples) collected from different retail stores of Rio Cuarto city in Argentina using HPLC-MS/MS. Sixty-seven samples (74%) showed contamination by fumonisins. From these samples, 16 showed fumonisin levels between LOD and LOQ (between 0.01 to 0.05 ng/g), while fumonisins (FB 1 + FB 2 ) in quantifiable samples ranged from 0.05 ng/g to 18.9 ng/g. Although FB 1 was more prevalent, FB 2 was foun3d in higher levels than FB 1 . Overall, fumonisin prevalence was high, but concentrations were far below EU or USA limits set for maize and maize-based products.

Mycotoxins are secondary metabolites produced by various molds that contaminate many staple foods and cause a broad range of detrimental health effects in animals and humans through chronic exposure or acute toxicity. As such, the worldwide contamination of food and feed with mycotoxins is a significant problem, especially in sub-Saharan Africa. In this study, mycotoxin occurrence in staple foods consumed in Somalia was determined. A total of 140 samples (42 maize, 40 sorghum, and 58 wheat) were collected from a number of markets in Mogadishu, Somalia, and analyzed by a UPLC-MS/MS multimycotoxin method that could detect 77 toxins. All of the maize samples tested contained eight or more mycotoxins, with aflatoxin B 1 (AFB 1 ) and fumonisin B 1 (FB 1 ) levels reaching up to 908 and 17 322 μg/kg, respectively, greatly exceeding the European Union limits and guidance values. The average probable daily intake of fumonisins (FB 1 and FB 2 ) was 16.70 μg per kilogram of body weight (kg bw) per day, representing 835% of the recommended provisional maximum tolerable daily intake value of 2 μg/(kg bw)/day. A risk characterization revealed a mean national margin of exposure of 0.62 for AFB 1 with an associated risk of developing primary liver cancer estimated at 75 cancers per year per 100 000 people for white-maize consumption alone. The results clearly indicate that aflatoxin and fumonisin exposure is a major public-health concern and that risk-management actions require prioritization in Somalia.

Fusarium temperatum is an emerging maize pathogen that causes maize ear and stalk rot diseases and produces various mycotoxins including moniliformin, beauvericin, enniatins and fumonisin B1, which poses a potential risk to the human food or animal feed supply chains. Early detection of F. temperatum is crucial to prevent its derived mycotoxins from entering the food chain, and is also a useful tool in disease management practices. Here, we describe a loop-mediated isothermal amplification (LAMP) assay for rapid diagnosis of F. temperatum. The 28S ribosomal DNA sequences (28S rDNA) of F. temperatum were used to design a set of six primers. The reaction conditions were optimized for developing a fast assay with high specificity and sensitivity, and were able to detect the presence of less than 10 pg of target DNA per reaction within 60 min. Furthermore, the resulting amplicons were visualized by adding SYBR Green I to the reaction tubes. Suspected F. temperatum infected maize stalk samples collected from Yunnan province, China were identified using the developed LAMP assay. In conclusion, the method not only provides a rapid and specific screening for the existence of F. temperatum in a bulk of maize samples without using sophisticated equipment, but also is potentially useful for other agriculturally important toxigenic fungi., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this study, 120 silage samples collected in 2015 from farms in Poland were analysed by a multimycotoxin method based on liquid chromatography coupled with tandem mass spectrometry. The study included toxins which are regulated within the European Union (aflatoxins, deoxynivalenol, fumonisins, T-2/HT-2 toxins, ochratoxin A and zearalenone) and non-regulated mycotoxins (enniatins, beauvericin, 8-ketotrichothecenes, sterigmatocystin, zearalenone derivatives). All silage samples were positive for at least one mycotoxin, and 61% of samples contained five or more mycotoxins simultaneously. The most frequently detected toxins were deoxynivalenol, nivalenol, zearalenone, enniatins and beauvericin, although the levels of these toxins were relatively low. The mean concentration of deoxynivalenol and zearalenon was 406 and 80.6 μg/kg, respectively, and two toxins were positive-correlated. This is the first study that provides information about emerging mycotoxins contaminating silage in Poland.

The influence of processing methods used to produce traditional Nigerian infant foods (ogi and processed soybean powder) on four European Union regulated Fusarium mycotoxins using naturally and artificially contaminated raw materials was studied using liquid chromatography-tandem mass spectrometry. Generally, there was a significant reduction of all the mycotoxins when compared to the initial concentration of the raw materials. Reduction in concentrations of the mycotoxins during ogi-processing started immediately after 36 h' steeping/fermentation for all the mycotoxins (fumonisin B 1 , zearalenone, deoxynivalenol, and T-2 toxin), and proceeded along the process chain (milling and sieving). In addition, deoxynivalenol-3-glucoside (16 ± 3.2 μg/kg) and 3-acetyl-deoxynivalenol (9 ± 5.5 μg/kg) initially absent in the raw maize were detected in the final ogi product. β-zearalenol, hydrolysed fumonisin B 1 , and HT-2 toxin were also detected at varying concentrations. Regarding soybean processing, a similar trend was observed with fumonisin B 1 , zearalenone, deoxynivalenol, and T-2 toxin, irrespective of the method used or the initial concentration. Other mycotoxins detected in soybean product include 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, deoxynivalenol-3-glucoside, HT-2 toxin, neosolaniol, α-zearalenol, β-zearalenol, and zearalenone-14-glucoside. Although there was a reduction in the concentration of the free mycotoxin because of processing, other mycotoxins were detected in the products and thus, may present an additional health risk on consumers., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Consumption of fumonisin-contaminated foods has a negative influence on the health of humans (carcinogen; oesophageal cancer in Eastern Cape in South Africa). Lactic acid bacteria (LAB) have emerged as a promising natural detoxification agent against mycotoxins. The aim of this study was to visualise the interaction between fumonisins (FB 1 and FB 2 ) and LAB: Lactobacillus plantarum FS2, L. delbrueckii subsp. delbrueckii CIP 57.8T and Pediococcus pentosaceus D39, isolated from traditional fermented maize-based products (ogi and mahewu) using confocal laser scanning microscopy (CLSM) and to then quantify the LAB-bound fumonisin using high performance liquid chromatography (HPLC). The objective was to obtain a physically visible and quantifiable binding interaction between fumonisins and LAB strains with the aim of utilising LAB as a possible detoxifying agent. Fumonisins were derivatised using naphthalene-2,3-dicarboxaldehyde (NDA) and then combined with non-fluorescent LAB cells (viable and non-viable). For the quantification of bound fumonisins, viable and non-viable cells were incubated in the presence of predetermined concentrations of fumonisins and the level of fumonisin in the suspension was determined. CLSM showed the derivatised green fluorescent fumonisins binding to the surface of each of the LAB cells. For viable cells, L. plantarum FS2 bound FB 1 most effectively while P. pentosaceus D39 bound the least level of FB 1 . The highest levels of FB 2 were bound by L. plantarum R 1096 and the least by L. delbrueckii CIP 57.8 T. For non-viable cells, L. plantarum FS2 was also the most effective for binding both fumonisins with P. pentosaceus D39 and L. delbrueckii CIP 57.8 T being the least effective for FB 1 and FB 2 , respectively. To our knowledge, this is the first study to visualise the interaction between LAB and fumonisins. We demonstrate that LAB isolates from indigenous fermented maize-based beverages bind fumonisins and thus present a potential strategy for their reduction in these traditional foods.

Fumonisins are a group of toxic secondary metabolites that are produced by Fusarium verticillioides which are associated with poultry health hazard and great economic losses. The objective of the present study was to develop an immunological method to detect F. verticillioides in poultry feed samples. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a polyclonal antibody against 67 kDa protein of the F. verticillioides 97K exoantigen was developed to detect this fungus. Antibody anti-67 kDa protein showed cross-reactivity against F. graminearum (2⁻7%) and F. sporotrichioides (10%), but no or low cross-reactivity against Aspergillus sp. and Penicillium sp. exoantigens. The detection limit for the 67 kDa protein of F. verticillioides was 29 ng/mL. Eighty-one poultry feed samples were analyzed for Fusarium sp. count, 67 kDa protein of F. verticillioides and fumonisin concentrations. Eighty of the 81 feed samples (98.6%) showed Fusarium sp. contamination (mean 6.2 x 10⁴ CFU/g). Mean 67 kDa protein and fumonisin concentration in the poultry feed samples was 21.0 µg/g and 1.02 µg/g, respectively. The concentration of 67 kDa protein, as determined by ic-ELISA correlated positively (p < 0.05) with fumonisin levels (r = 0.76). These results suggest that this ic-ELISA has potential to detect F. verticillioides and predict fumonisin contamination in poultry feed samples.

A multiplex Lateral Flow Immunoassay was developed based on the use of a single Test line and multicolour gold nanoparticles (GNPs) as signal reporters. Red and blue GNPs were linked to antibodies directed towards two different analytes and included in a typical lateral flow immunoassay configuration, in which the Test line was formed by the mixture of two antigens. As a result of the immunoreactions occurring at the Test zone, diverse combinations of red and blue GNPs labels were captured. Therefore, the Test line assumed different colours depending on which - and how much - analyte is present in the sample. The multiplexing capability of the 'colour-encoded assay' is illustrated by the simultaneous detection of aflatoxin B1 (AFB1) and type-B fumonisins (FMs) in wheat and food products that made with wheat. Reproducible detection of AFB1 and FMs contamination in raw and processed food was achieved with visual cut-off levels at 1 ng mL -1 and 50 ng mL -1 , respectively. The contaminant was identified based on the colour of the label according with a specific colour code. Furthermore, strips images were acquired by means of a common smartphone and analysed through RGB data analysis providing semi-quantitative detection of the two mycotoxins., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Systemic infection through the seed is one of the routes used by the mycotoxinogenic pathogen Fusarium verticillioides for colonizing maize plants. The prohibition of the use of most chemical fungicides by the EU has promoted research on plant resistance inducers as an effective and sustainable alternative. Induction of a priming state in maize seeds might affect their susceptibility to contamination and accumulation of fumonisins. This state by application of a natural fertilizer called Chamae on maize seeds, was investigated in two varieties to control the colonization by the fungus and the accumulation of fumonisins B 1 , B 2 and B 3 , germinating seeds, dead plants and yield., Results: After inoculation of F. verticillioides on germinating seeds, the colonization by the fungus and the accumulation of fumonisins were significantly lower in seedlings coming from treated seeds, but a significant number of plants stopped their development by necrosis. In a field trial, the 0.01% (v/v) application dilution showed a lower plant density, although the level of biomass at harvest was not affected., Conclusion: The priming state contributed to the control of F. verticillioides development from seed infection and fumonisin accumulation in the early stage of plant growth, without affecting the final crop yield, and could reduce fungicide use and environmental contamination. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Brazil is one of the world's largest corn producers and is a leader in exportation. Due to intense globalization, corn may be commercialized worldwide and the issue concerning the safety of corn-based products has become a topic of widespread international interest. Dietary exposure evaluation is a relevant criterion for mycotoxin risk assessment. Thus, human exposure to fumonisins were assessed for corn grain and its derivatives (endosperm, cornmeal, and grits; n = 320) sampled from one of the large-scale corn processing plants in Brazil. The total probable daily intake (PDI) for fumonisins in Brazil was 96.9 ng kg -1 body weight day -1 , which corresponds to 5% of the provisional maximum tolerable daily intake (PMTDI) of 2000 ng kg -1 b.w. day -1 for fumonisins. In countries that import Brazilian corn, the total PDI is lower in European countries (from 35.7 to 177 ng kg -1 b.w. day -1 ) and higher in Angola (1553 ng kg -1 b.w. day -1 ). Taking into account that dietary exposure in populations in Brazil and importing countries was low, the corn-based products were safe for human consumption regarding fumonisins, even for regions with high corn consumption.

Deoxynivalenol (DON), Fumonisin B 1 (FB 1 ) and Aflatoxin B 1 (AFB 1 ) have strong toxicity to humans and exist widely in grain and food. It is necessary to screen these mycotoxins to guarantee food safety. In order to develop a rapid, simple, low-cost and simultaneous-detection method, composites of antibody-nano-Au-particles, DON-BSA, FB 1 -BSA, and AFB 1 -BSA were prepared to establish a lateral flow assay based on competitive inhibition. The results suggested that the visual detection limits for DON, FB 1 and AFB 1 were 10, 30, and 10 ng mL -1 , respectively. On the other hand, it had no reactivity to T-2 toxin, zearalenone, ochratoxin A and nivalenol, which demonstrates good specificity. Besides, the strip had good repeatability and stability. Therefore, DON, FB 1 and AFB 1 would be screened simultaneously by lateral flow assay for food safety., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The fungus Fusarium verticillioides can infect maize ears, contaminating the grain with mycotoxins, including fumonisins. This global public health threat can be managed by breeding maize varieties that are resistant to colonization by F. verticillioides and by sorting grain after harvest to reduce fumonisin levels in food systems. Here, we employed two F. verticillioides inoculation techniques representing distinct infection pathways to dissect ear symptomatology and morphological resistance mechanisms in a diverse panel of maize inbred lines. The "point" method involved penetrating the ear with a spore-coated toothpick and the "inundative" method introduced a liquid spore suspension under the husk of the ear. We evaluated quantitative and qualitative indicators of external and internal symptom severity as low-cost proxies for fumonisin contamination, and found that kernel bulk density was predictive of fumonisin levels (78 to 84% sensitivity; 97 to 99% specificity). Inundative inoculation resulted in greater disease severity and fumonisin contamination than point inoculation. We also found that the two inoculation methods implicated different ear tissues in defense, with cob morphology being a more important component of resistance under point inoculation. Across both inoculation methods, traits related to cob size were positively associated with disease severity and fumonisin content. Our work demonstrates that (i) the use of diverse modes of inoculation is necessary for combining complementary mechanisms of genetic resistance, (ii) kernel bulk density can be used effectively as a proxy for fumonisin levels, and (iii) trade-offs may exist between yield potential and resistance to fumonisin contamination.

In this report, a UPLC-ESI-MS/MS method for the simultaneous determination of aflatoxins, ochratoxin A, zearalenone, deoxynivalenol, fumonisins, T-2 and HT-2 toxins, fusarenone X, diacetoxyscirpenol, and 3- and 15-acetyldeoxynivalenol in feedstuffs was developed. A quadrupole-time-of-flight mass spectrometer detector (QTOF-MS) operating in full scan mode was combined with the UPLC-ESI-MS/MS system to confirm the identity of detected mycotoxins and to identify other possible microbial metabolites occurring in samples. Sixty-two feed samples from the Spanish market were analyzed. Extraction of metabolites was carried out with acetonitrile-water-formic acid (80:19:1, v/v/v). Method detection and quantification limits and performance criteria set by Commission Regulation (EC) No 401/2006 were fulfilled. Relatively high levels of the main regulated mycotoxins and presence of non-regulated mycotoxins in feed samples were found. This is the first study in which mycotoxins and other microbial metabolites occurring in feed are studied using a UPLC-QTOF-MS system being therefore a reference report., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Homogeneous immunoassays represent an attractive alternative to traditional heterogeneous assays due to their simplicity, sensitivity, and speed. On the basis of a previously identified epitope-mimicking peptide, or mimotope, we developed a homogeneous fluorescence quenching immunoassay based on gold nanoparticles (AuNPs) and a recombinant epitope-mimicking fusion protein for the detection of mycotoxin fumonisin B 1 (FB 1 ). The fumonisin mimotope was cloned as a fusion protein with a yellow fluorescent protein that could be used directly as the tracer for FB 1 detection without the need of labeling or a secondary antibody. Furthermore, owing to the fluorescence quenching ability of AuNPs, a homogeneous immunoassay could be performed in a single step without washing steps to separate the unbound tracer. The homogeneous quenching assay showed negligible matrix effects in 5% wheat extract and high sensitivity for FB 1 detection, with a dynamic range from 7.3 to 22.6 ng mL -1 , a detection limit of 1.1 ng mL -1 , and IC 50 value of 12.9 ng mL -1 , which was significantly lower than the IC 50 value of the previously reported assay using the synthetic counterpart of the same mimotope in a microarray format. The homogeneous assay was demonstrated to be specific for fumonisins B 1 and B 2 , as no significant cross-reactivity with other mycotoxins was observed, and acceptable recoveries (86% for FB 1 2000 μg kg -1 and 103% for FB 1 4000 μg kg -1 ), with relative standard deviation less than 6.5%, were reported from spiked wheat samples, proving that the method could provide a valuable tool for simple analysis of mycotoxin-contaminated food samples.

The stability of the Fusarium mycotoxins fumonisin B 1 , deoxynivalenol, T-2 toxin, and zearalenone during processing of Nigerian traditional spices (dawadawa, okpehe, and ogiri) and beer (burukutu) using artificially contaminated raw materials was investigated. Results revealed the reduction of these toxins in all the final products. Boiling played a significant role (p < 0.05) in Fusarium mycotoxin reduction in the traditional spices. The highest percentage reduction of deoxynivalenol (76%) and zearalenone (74%) was observed during okpehe processing (boiled for 12 h). Dehulling and fermentation further demonstrated a positive influence on the reduction of these toxins with a total reduction ranging from 85 to 98% for dawadawa, 86 to 100% for okpehe, and 57 to 81% for ogiri. This trend was also observed during the production of traditional beer (burukutu), with malting and brewing playing a major impact in observed reduction. In addition, other metabolites including deoxynivalenol-3-glucoside, 15-acetyl-deoxynivalenol, α-zearalenol, and β-zearalenol which were initially not present in the raw sorghum were detected in the final beer product at the following concentrations 26 ± 11, 16 ± 7.7, 22 ± 18, and 31 ± 16 μg/kg, respectively. HT-2 toxin was also detected at a concentration of 36 ± 13 μg/kg along the processing chain (milled malted fraction) of the traditional beer. For the traditional spices, HT-2 toxin was detected (12 μg/kg) in ogiri. Although there was a reduction of mycotoxins during processing, appreciable concentrations of these toxins were still detected in the final products. Thus, the use of good quality raw materials significantly reduces mycotoxin contamination in final products.

One of the food security problems faced worldwide is the occurrence of mycotoxins in grains and their foods. Fumonisins (FBs) are mycotoxins which are prevalent in corn (Zea mays L.) and its based foods. Their intake and exposure have been epidemiologically and inconclusively associated with oesophageal cancer and neural tube defects in humans, and other harmful health effects in animals. The toxic effects of FBs can be acute or chronic and these metabolites bioaccumulate mainly in liver and kidney tissues. Among FBs, fumonisin B 1 (FB 1 ) is the most relevant moiety although the 'hidden' forms produced after food thermal processes are becoming relevant. Corn is the grain most susceptible to Fusarium and FBs contamination and the mould growth is affected both by abiotic and biotic factors during grain maturation and storage. Mould counts are mainly affected by the grain water activity, the environmental temperature during grain maturation and insect damage. The abiotic factors affected by climatic change patterns have increased their incidence in other regions of the world. Among FBs, the hidden forms are the most difficult to detect and quantify. Single or combined physical, chemical and biological methods are emerging to significantly reduce FBs in processed foods and therefore diminish their toxicological effects.

Colorimetric detection has been widely applied in daily life. However, on the other hand, the further development of colorimetric biosensors has been constrained by the lack of multiple detection capabilities. In this work, we proposed a pH-resolved colorimetric biosensor for the first time for the simultaneous detection of four targets, including ochratoxin A (OTA), aflatoxins B1 (AFB1), fumonisin B1 (FB1), and microcystin-LR (MC-LR). With allochroic dyes as the signal indicators, pH value was introduced as the new dimension to control the release of dyes. The concentrations of four targets can be obtained in order by adjusting the pH of the solution. Meanwhile, tedious and high-cost chemical modification processes in the fabrications of biosensors were also avoided by the combination of DNA-directed self-assembly of graphene oxide and magnetic separation. This biosensor provided a simple, rapid, accurate, and low-cost strategy for multiple target detection.

The aim of this study was to evaluate the potential use of an e-nose in combination with lateral flow immunoassays for rapid aflatoxin and fumonisin occurrence/co-occurrence detection in maize samples. For this purpose, 161 samples of corn have been used. Below the regulatory limits, single-contaminated, and co-contaminated samples were classified according to the detection ranges established for commercial lateral flow immunoassays (LFIAs) for mycotoxin determination. Correspondence between methods was evaluated by discriminant function analysis (DFA) procedures using IBM SPSS Statistics 22. Stepwise variable selection was done to select the e-nose sensors for classifying samples by DFA. The overall leave-out-one cross-validated percentage of samples correctly classified by the eight-variate DFA model for aflatoxin was 81%. The overall leave-out-one cross-validated percentage of samples correctly classified by the seven-variate DFA model for fumonisin was 85%. The overall leave-out-one cross-validated percentage of samples correctly classified by the nine-variate DFA model for the three classes of contamination (below the regulatory limits, single-contaminated, co-contaminated) was 65%. Therefore, even though an exhaustive evaluation will require a larger dataset to perform a validation procedure, an electronic nose (e-nose) seems to be a promising rapid/screening method to detect contamination by aflatoxin, fumonisin, or both in maize kernel stocks.

A label-free optical biosensor for the fast simultaneous determination of three mycotoxins, aflatoxin B 1 (AFB 1 ), fumonisin B 1 (FB 1 ) and deoxynivalenol (DON), in beer samples is presented. The biosensor is based on an array of ten Mach-Zehnder interferometers (MZIs) monolithically integrated along with their respective broad-band silicon light sources onto a single chip. Multi-analyte determination is accomplished by functionalizing the sensing arms of individual MZIs with mycotoxin-protein conjugates. Assay is performed by pumping over the chip mixtures of calibrators or samples with a mixture of specific monoclonal antibodies, followed by reaction with a secondary anti-mouse IgG antibody. Reactions are monitored in real-time by continuously recording the MZI output spectra, which are then subjected to Discrete Fourier Transform to convert spectrum shifts to phase shifts. The detection limits achieved for AFB 1 , FB 1 and DON were 0.8, 5.6 and 24 ng/ml, respectively, while the assay duration was 12 min. Recovery values ranging from 85 to 115% were determined in beer samples spiked with known concentrations of the three mycotoxins. In addition, beers of different types and origin were analysed with the biosensor developed and the results were compared with those provided by established laboratory methods, further supporting the accuracy of the proposed device., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Mycotoxins are secondary metabolites of fungi that are damaging to both animals and humans. Extensive contamination of foods and feeds with mycotoxins is an important problem. Fumonisins, trichothecenes, zearalenone, and aflatoxins are mycotoxins produced by Fusarium species and occur naturally in sugarcane and cereal-based foods, threatening health and food security worldwide. Their distribution in the contaminated material is of great interest for obtaining insight into infection mechanisms and the potential for reducing contamination during food processing. In this study, mycotoxins were evaluated by high-performance liquid chromatography, liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), and matrix-assisted laser/desorption ionization time of flight mass spectrometry (MALDI-TOF MS) of Fusarium species-infected sugarcane materials. A simple, sensitive, and reliable analytical method was developed for rapidly detecting eight mycotoxins in Fusarium species: fumonisin B1 and fumonisin B2, B-trichothecene mix (deoxynivalenol, nivalenol, 3-acetyl- deoxynivalenol, 15-acetyl-deoxynivalenol), zearalenone, and aflatoxin G1. Analyses were carried out in multiple reaction monitoring mode using the two primary product ions. The results generated by LC/MS and MALDI-TOF MS/MS revealed various mechanisms regulating mycotoxins production, which may help to clarify the roles of sensitive and selective compounds. The results demonstrate that this procedure is suitable for simultaneous determination of mycotoxins in sugarcane and can be performed in routine analysis in mycotoxin laboratories., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

In temperate areas, the main limitation to the use of maize in the food chain is its contamination by B-series fumonisins (FBs) during cultivation. Since the content of this group of mycotoxins may be distributed unevenly after milling, the aim of this study was to compare the distribution of FBs in maize fractions derived from two industrial dry-milling processes, that is, a dry-degermination (DD) system and a tempering-degermination (TD) system. Grain cleaning reduces FBs by about 42%. The germ of the two degermination processes showed a similar FB content of kernel after cleaning. Conversely, an animal feed flour resulted in a FB content that was two times higher than whole grain before cleaning. A significant FB reduction was observed in the milling fractions in both processes, with a higher reduction in the TD system than in the DD one. The average decontamination respect to uncleaned kernels in the DD process was 50%, 83% and 87%, for maize flour, break meal and pearl meal, respectively, while it was 78%, 88% and 94% in the TD process for small, medium and flaking grits, respectively. Among the milling fractions, the flaking grits with the highest particle size resulted in the highest FB reduction.

Naked-eye readout colorimetric signal-based assays are promising for high-throughput screening detection and point-of-care diagnostics in resource-constrained regions. Here, a novel direct competitive plasmonic enzyme-linked immunosorbent assay (dc-pELISA) based on gold nanoparticle (AuNP) aggregation with highly sensitive and robust naked-eye readout signal was developed and used to detect fumonisin B 1 (FB 1 ). AuNP aggregation was induced by a horseradish peroxidase (HRP)/hydrogen peroxide (H 2 O 2 )/tyramine (TYR) system, resulting in a dramatic color change from red to blue. In this system, H 2 O 2 was produced via GOx-mediated glucose oxidation reaction, and FB 1 -GOx was used as a competitive antigen. The proposed pELISA demonstrated good linear detection of FB 1 from 3.125 ng mL -1 to 25 ng mL -1 with a vivid color change from deep blue to red and a cutoff limit of 12.5 ng mL -1 observed by the naked eye. The average recoveries for FB 1 -spiked corn samples ranged from 76.5% to 96.8% with a relative standard derivation of 4.88~ 16.4%. Meanwhile, the proposed method exhibited excellent agreement (R 2 = 0.927) with the conventional ELISA method in blindly detecting FB 1 spiked corn samples. Additionally, the results of the proposed method for FB 1 positive corn samples also showed a high consistency with those of the ultra performance liquid chromatography method. These results indicated acceptable accuracy and precision of the proposed colorimetric ELISA for quantitative detection of FB 1 in actual corn samples., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This paper presents data on the occurrence of Fusarium toxins - zearalenone (ZEA), deoxynivalenol (DON) and fumonisins (FUMs) B1 and B2 - in corn flours and corn flakes marketed in Serbia. A total of 71 samples were collected over 2013-2016 and analysed using HPLC with UV or fluorescence detection. In the case of corn flours, none of the samples taken in 2013 exhibited the presence of ZEA or DON, whereas 90% were positive for FUMs. In 2015, occurrence was very high: ZEA 93%, DON 86% and FUMs 100% (mean 43.3, 322.6 and 323.0 μg kg -1 , respectively), with 21% of the samples exceeding the maximum level for ZEA and 7% for DON and FUMs. In 2016, a lower occurrence was recorded in the case of ZEA (75%) and DON (38%), with drastically lower mean contamination levels (six- and 10-fold, respectively), while FUMs stayed at 97%, with twofold lower mean. The maximum level was exceeded only for ZEA (3%). The frequency of ZEA-DON-FB1 co-occurrence was 86% in 2015 and 25% in 2016. Regarding corn flakes, occurrence summarised for the study period was 87% ZEA, 73% FUMs and 40% DON. One sample (7%) exceeded the maximum levels for both ZEA and DON. Observed occurrence changes were in agreement with the climatic conditions during corn growing seasons preceding the market release of the processed products.

The enzyme-linked immunosorbent assay (ELISA) has been used as a standard tool for monitoring food and animal feed contamination from the carcinogenic fumonisin B1 (FB1). Unfortunately, ELISA is not always efficient due to the instability of the antibody and enzyme components in the immunoassay, the presence of natural enzyme inhibitors in the samples and the high levels of non-specific protein binding. Additionally, the production of antibodies for ELISA can be time-consuming and costly, due to the involvement of animals in the manufacturing process. To overcome these limiting factors, a molecularly imprinted nanoparticle based assay (MINA) has been developed, where the molecularly imprinted nanoparticles (nanoMIPs) replace the primary antibody used in a competitive ELISA. Herein, computational modelling was used to design the nanoMIPs by selecting monomers that specifically interact with FB1. The affinity of the monomers to FB1 was verified by measuring their binding in affinity chromatography experiments. The nanoMIPs were produced by solid phase synthesis and the results showed that nanoMIPs had a hydrodynamic diameter of around 249 ± 29 nm. The assay tested in model samples is highly selective and does not show cross-reactivity with other mycotoxins such as fumonisin B2 (FB2), aflatoxin B1 (AFB1), citrinin (CTT), zearalenone (ZEA), and deoxynivalenol (DON). The MINA allows the detection of FB1 in the concentration range of 10 pM-10 nM with a detection limit of 1.9 pM and a recovery of 108.13-113.76%.

Background: In the present study, a novel highly sensitive magnetic enzyme chemiluminescence immunoassay (MECLIA) was developed to detect fumonisin B 1 (FB 1 ) in cereal samples. The gold-coated magnetic nanoparticles (Fe 3 O 4 @Au, GoldMag) were used as solid phase carrier to develop a competitive CLIA for detecting FB 1 , in which FB 1 in samples would compete with FB 1 -ovalbumin coated on the surface of Fe 3 O 4 @Au nanoparticles for binding with FB 1 antibodies. Successively, horseradish peroxidase labeled goat anti-rabbit IgG (HRP-IgG) was conjugated with FB 1 antibodies on the microplate. In substrate solution containing luminol and H 2 O 2 , HRP-IgG catalyzed luminol oxidation by H 2 O 2 , generating a high chemiluminescence signal. The FB 1 immune GoldMag particles were characterized by Fourier transform infrared spectroscopy, scanning electron microscope and zeta potential analysis, etc. RESULTS: The concentrations and the reaction times of these immunoreagents were optimized to improve the performances of this method. The established method could detect as low as 0.027 ng mL -1 FB 1 from 0.05 ng mL -1 to 25 ng mL -1 , demonstrating little cross-reaction (less than 2.4%) with other structurally related compounds. The average intrassay relative SD (RSD) (n = 6) was 3.4% and the average interassay RSD (n = 6) was 5.4%. This method was successfully applied for the determination of FB 1 in corn and wheat and gave recoveries of between 98-110% and 91-105%, respectively., Conclusion: The results of the present study suggest that the MECLIA approach has potential application for high-throughput fumonisin screening in cereals. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Fungicides and, for the first time, microalgal phenolic extracts (MPE) from Spirulina sp. and Nannochloropsis sp. were applied on maize culture media under field conditions to evaluate their ability to minimize Fusarium species development and fumonisin production. An in vitro assay against F. verticillioides was carried out using maize grains as the culture medium. An open-field experiment was carried out in Northwest Italy under natural infection conditions. The compared treatments were factorial combinations of two insecticide treatments (an untreated control and pyrethroid, used against European Corn Borer), four antifungal treatments (an untreated control, MPE from Spirulina sp., MPE from Nannochloropsis sp., and a synthetic fungicide), and two timings of the application of the antifungal compounds (at maize flowering and at the milk stage). The MPE compounds were capable of inhibiting fumonisin production in vitro more efficiently than tebuconazole. Insecticide application reduced the infection by Fusarium species and subsequent fumonisin contamination. However, fumonisins in maize fields were not significantly controlled by either fungicide or MPE application.

Maize ( Zea mays) is a staple in many developing countries but is known to be prone to pest (insects, birds, and rodents) and fungal infestation. In Guatemala, mycotoxin contamination of cultivated products may occur owing to such factors as environmental conditions and the use of traditional agriculture operations. To assess the current maize conditions in Guatemala, a small-scale study was performed. Mold and insect counts and mycotoxin (aflatoxin and fumonisin) concentrations were determined on 25 farms in two townships (Chiantla and Todos Santos) of the Huehuetenango Department. Total fungal counts were 3.6 to 6.83 log CFU/g with no significant differences ( P > 0.05) across farms at different altitudes. Farms where maize was not produced but was purchased were at higher risk of fumonisin contamination, whereas local producers were mostly affected by aflatoxins. Aflatoxin was present in maize from 100% of farms at 1.0 to 85.3 ppb, and fumonisin was detected on 52% of farms at 0.4 to 31.0 ppm. Average mycotoxin consumption amounts were above the recommended maximum intake for aflatoxin in both produced and purchased maize and above the provisional maximum tolerable daily intake for fumonisin in purchased maize. Estimated daily intake was 0.01 to 0.85 μg/kg of body weight per day for aflatoxin and 2.9 to 310.0 μg/kg of body weight per day for fumonisin. An entomological analysis revealed overall 32% prevalence of Ephestia kuehniella (flour moth), 16% prevalence of Sitophilus zeamais (maize weevil), and 8% prevalence of Tribolium sp. (flour beetle) on the analyzed farms. This study highlighted poor agricultural practices used in the highlands of Guatemala. Current practices should be revised for the production of maize that is safe for consumption by the population in this region.

A visual immunoaffinity test column (IATC) assay was developed to detect fumonisins in cereal samples for spot tests without the need for special instruments. The developed IATC assay had equivalent recognition capability for fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), or fumonisin B₃ (FB₃), and exhibited no cross-reactivity with aflatoxin B₁, ochratoxin A, zearalenone, or the T-2 toxin. The sample pretreatment was accomplished more rapidly and with greater ease, the entire assay procedure was completed in approximately 10 min, including sample pretreatment and testing. The limits of detection (LODs) of the IATC assay to detect fumonisins in the maize, barley, oat, and millet samples were 20 &mu;g kg &minus;1 . The results of the spiked maize, barley, oat, and millet and real maize samples by the IATC assay agreed well with the results obtained by the commercial fumonisin enzyme-linked immunosorbent assay (ELISA) test kit and liquid chromatography-tandem mass spectrometry (LC-MS/MS), respectively. The developed IATC assay can serve as a useful screening tool for the rapid, qualitative, and semi-quantitative detection of the total content of fumonisins (sum of FB₁, FB₂, and FB₃) in cereal samples on-site.

Here, we for the first time used a thiolated amino-ligand modified multi-branched gold nanoflower as skeleton to encapsulate iron porphyrins (AuNF@FeTPPCl) as alternatives to horseradish peroxidase (HRP). After the FeTPPCl encapsulation, the HRP mimicking activity of FeTPPCl was effectively blocked by the steric hindrance of the hydrophobic layer on the AuNF surface. Upon the addition of ethanol, the loaded FeTPPCl was released into the solution in its free format and exposed the catalytic sites, resulting in the recovery of catalytic activity. The proposed encapsulation method effectively avoided the loss of catalytic activity that originated from the blocking of the catalytic active sites during immobilization. Additionally, the resultant AuNF@FeTPPCl nanocomposite exhibited a high loading level with 2.4 × 10 6 FeTPPCl molecules per AuNF, and showed considerably high catalytic activity for hydrogen peroxide and 3, 3' 5, 5'-tetramethylbenzidine, which is approximately 40- and 172-folds higher than native HRP. Through using the as-prepared AuNF@FeTPPCl as an alternative of HRP for trace labeling, we successfully developed colorimetric immunosensors for fumonisin B 1 (FB 1 ) and hepatitis B surface antigen (HBsAg) with competitive and sandwich-type formats, respectively. The developed AuNF@FeTPPCl-based colorimetric immunosensor exhibited higher detection sensitivity for FB 1 and HBsAg than the corresponding HRP-based immunosensors. Thus, the proposed AuNF@FeTPPCl can be used as HRP mimicking analogs for developing highly sensitive colorimetric immunosensor and for loading other hydrophobic iron porphyrins or catalysts., (Copyright © 2017. Published by Elsevier B.V.)

Beer, a beverage consumed throughout the world, is mainly derived from cereals. In this study, fungal and mycotoxin contamination, as well as the physicochemical properties of maize-based opaque beer (umqombothi) obtained from the Gauteng province of South Africa, was investigated. The mean water activity, pH and total titratable acidity of the analysed beer samples were 0.91, 3.76 and 1.20% lactic acid, respectively. The investigation revealed Aspergillus, Penicillium, Phoma and Saccharomyces as the predominant fungal genera with a mean fungal load of 3.66 × 10 5  CFU/mL. Among the mycotoxigenic fungal species recovered, Aspergillus flavus had the highest incidence of 26%. Previously unreported strains such as P. chrysogenum strain AD25, A. sydowii strain AD 22 and A. tritici strain AD 11 were found. Furthermore, mycotoxin quantitative analysis via liquid chromatography-tandem mass spectrophotometry showed that deoxynivalenol was the dominant mycotoxin occurring in 84% of the samples. This was followed by enniatin B that occurred in 75% of samples ranging from 12 to 44 μg/L and fumonisin B 1 (FB 1 ) (incidence of 53% at a maximum level of 182 μg/L). Generally, there was low occurrence aflatoxins, whereas T-2, HT-2, nivalenol, zearalenone, 3- and 15-acetyl-deoxynivalenol were not detected. All the samples analysed had safe levels of mycotoxins tested but were contaminated by at least two mycotoxins that could pose some additive or synergistic health effects among consumers. On average: a 60 kg adult consuming 1-6 L/day of the beer was exposed to FB 1  + FB 2 at an estimated 2.20-13.20 μg/kg body weight/day. These values were far above the maximum tolerable daily intake of 2 μg/kg bw/day established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). The study demonstrates that consumption of umqombothi can significantly enhance dietary exposure to multiple mycotoxins among consumers, and therefore accentuates the need for strategies aimed at reducing toxigenic fungal colonization and mycotoxin contamination in the beer processing chain., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study investigated the presence of Aspergillus species belonging to Aspergillus section Nigri on Vitis labrusca and its hybrid grapes grown in Brazil. The ability of the fungi isolates to produce ochratoxin A (OTA) and fumonisin B 2 (FB 2 ) as well as the presence of these mycotoxins in the grapes were also studied. Eighty-eight samples were collected from the main grape producing states in Brazil: Rio Grande do Sul (n=30), Pernambuco (n=21), São Paulo (n=21) and Paraná (n=16). The highest average contamination level by A. section Nigri occurred on the grapes from Pernambuco (66.3%). A total of 2042 A. section Nigri isolates was analyzed and clustered in three groups according to morphology characterization: A. section Nigri uniseriate (79.3%), A. niger "aggregate" (18.3%) and A. carbonarius (2.4%). In order to precisely identify the Aspergillus species, two hundred and forty-eight strains were subjected to DNA sequencing. Among the A. section Nigri uniseriate group, the following species were found: A. japonicus, A. uvarum, A. brunneoviolaceus, A. aculeatus and A. labruscus. Within the A. niger "aggregate", the following species were found: A.niger sensu stricto, A. welwitschiae and A. vadensis. Regarding mycotoxin-production capacity, 3.2% of the total A. section Nigri isolates (2042) were positive for OTA production and from A. niger "aggregate" (373) tested, 42.1% were FB 2 producers. However, none of the 88 grape samples were contaminated with these mycotoxins., (Copyright © 2017. Published by Elsevier B.V.)

To assess the risk of fumonisin contamination in Korean cereals, we isolated colonies of the Fusarium fujikuroi species complex (FFSC) from barley, maize, rice and soybean samples from 2011 to 2015. A total of 878 FFSC strains were isolated mostly from maize and rice, and species identity of the isolates were determined using the DNA sequence of the translation elongation factor 1-α (TEF-1α) and RNA polymerase II (RPB2) genes. Fusaria recovered from Korean cereals included F. fujikuroi (317 isolates and a frequency of 36%), F. proliferatum (212 isolates and 24.1%), F. verticillioides (170 isolates and 19.4%), F. concentricum (86 strains and 9.8%), F. andiyazi (56 isolates and 6.4%), F. subglutinans (28 isolates and 3.2%), F. thapsinum (5 isolates and 0.6%), and F. circinatum (2 isolates and 0.2%). The rice samples were dominated by F. fujikuroi (47.4%), F. proliferatum (27.3%), and F. concentricum (15.1%), whereas maize samples were dominated by F. verticillioides (33.9%), F. fujikuroi (25.3%), and F. proliferatum (21.1%). A phylogenetic analysis of 70 representative isolates demonstrated that each species was resolved as genealogically exclusive in the ML tree. Fumonisin production potential was evaluated using a PCR assay for the fumonisin biosynthesis gene, FUM1 in all of the isolates. Most of the isolates tested (94%) were positive for FUM1. All of the isolates assigned to F. fujikuroi, F. proliferatum, F. verticillioides and F. thapsinum were positive for FUM1 irrespective of their host origin. Seventy-seven representative isolates positive for FUM1 were examined for fumonisin production in rice medium. The majority of F. proliferatum (26/27, 96.3%), F. verticillioides (16/17, 94.1%) and F. fujikuroi (19/25, 76.0%) produced both FB 1 and FB 2 . Notably, 16 of 19 fumonisin-producing F. fujikuroi produced >1000μg/g of fumonisins (FB 1 +FB 2 ) in rice medium, which is higher than that in previous reports. These results suggest that F. fujikuroi can produce high levels of fumonisins similar to F. verticillioides and F. proliferatum., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Fusarium verticillioides is the most common fungal pathogen associated with maize ear rot in Tanzania. In a two-year trial, we investigated the efficacy of crop protection (insecticide and/or fungicide) and fertilizer (nitrogen and/or phosphorus) treatments in reducing the occurrence of F. verticillioides and its mycotoxins in maize grown in Tanzania. Seasonal differences were seen to have a substantial influence on the incidence and severity of insect infestation, Fusarium ear and kernel rot, biomass of F. verticillioides and contamination with fumonisins. With regard to the application of fertilizers, it was concluded that the impact on maize stalk borer injury, Fusarium symptoms and fumonisin levels was not significant, whereas crop protection significantly reduced maize damage. The application of an insecticide was most effective in reducing insect injury and as a result of the reduced insect injury the insecticide treatment also resulted in a significant decrease in Fusarium symptoms. In 2014, fumonisin levels were also significantly lower in maize treated with an insecticide. Additionally, significant positive correlations between insect damage and Fusarium symptoms were observed. In conclusion, this study clearly shows that application of an insecticide alone or in combination with a fungicide at anthesis significantly reduces insect damage and consequently reduces F. verticillioides infection and associated fumonisin contamination., Competing Interests: The authors declare no conflict of interest.

This study was conducted to investigate the natural co-occurrence of 295 fungal and bacterial metabolites in 28 samples of dried date palm fruits collected from different shops distributed in Assiut Governorate, Upper Egypt in 2016. Extraction and quantification of the target analytes were done using the "dilute and shoot" approach followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. In total, 30 toxic fungal metabolites were detected. Among these metabolites, 4 types of ochratoxins including ochratoxin type A and B were quantified in 3 samples (11%) with a contamination range from 1.48 to 6070 μg/kg for ochratoxin A and from 0.28 to 692 μg/kg for ochratoxin B. In addition, fumonisin B 2 was observed in 2 (7%) samples with contamination levels ranging from 4.99 to 16.2 μg/kg. The simultaneous detection of fumonisin B 2 in the same contaminated samples with ochratoxins indicates the fungal attack by Aspergillus niger species during storage. Only 1 sample was contaminated with aflatoxin B 1 (14.4 μg/kg) and B 2 (2.44 μg/kg). The highest maximum concentration (90400 μg/kg) was for kojic acid that contaminated 43% of the samples. To the best of the authors' knowledge, this is the first report of the natural co-occurrence of fumonisin B 2 and ochratoxin A and B in addition to a wide range of other fungal metabolites in date palm fruits., Practical Application: Mycotoxins are secondary metabolites produced by different fungi. These metabolites pose a potential risk on human health since they contaminate many food commodities. Among these, date palm fruits which are an integral part of diet in several countries. Therefore, detection of mycotoxins is a prerequisite to insure the safety of food. Here, different types of mycotoxins have been detected in levels that may have health hazard., (© 2018 Institute of Food Technologists®.)

A lateral flow immunoassay (LFIA) was developed for the determination of fumonisin mycotoxins. The fluorescence of CdSe/ZnS quantum dots (QDs), observed at excitation/emission wavelengths of 365/525 nm, is suppressed by the addition of silver nanoparticles (SNPs) or gold nanoparticles (GNPs) because SNPs overlap the excitation bands of the QDs, and GNPs overlap the emission bands. The fluorescence of the QDs is recovered upon addition of fumonisins, allowing for the sensitive detection in "positive mode" of the target mycotoxin by monitoring the changes of the QDs fluorescence intensity. The SNPs are found to be the most effective quenchers, while the use of GNPs allows for an efficient recovery of fluorescence and can be employed in the LFIA. The method was successfully applied to the fluorometric determination of fumonisins in spiked maize flour samples. The visual detection limit is at the ng·mL -1 level. This is four times lower compared to the colorimetric LFIA based on the use of the conventional gold NPs. Graphical abstract Schematic of the fluorescence quenching lateral flow immunoassay that uses fluorescent quantum dots (QD) and metal nanoparticles (NP) as the quencher: the binding of NP-labelled antibody to the antigen (purple triangle) modulates QD luminescence at the Test line, allowing for 'positive mode' detection of fumonisins. The NP accumulation at Control line assures validity of the test.

We surveyed the presence of 22 mycotoxins in 191 Spanish cow milk samples. Mycotoxins could be carried over from diet into animal milk and have toxic effects on human and animal health. The interaction of different mycotoxins may be additive or synergetic. Therefore, surveillance of mycotoxin co-occurrence in milk is recommended. Aflatoxins M 1 , B 1 , B 2 , G 1 , and G 2 , ochratoxins A and B, nivalenol, deoxynivalenol, deepoxy-deoxynivalenol, 3- and 15-acetyldeoxynivalenol, diacetoxyscirpenol, neosolaniol, fusarenon X, T-2 and HT-2 toxins, fumonisins B 1 , B 2 , and B 3 , sterigmatocystin, and zearalenone were analyzed. Samples were treated by liquid-liquid extraction with acidified acetonitrile, followed by an acetonitrile-water phase separation using sodium acetate. The analysis was carried out by HPLC coupled to a triple quadrupole mass spectrometer. None of the analyzed mycotoxins had a concentration level higher than their detection limit (0.05-10.1 µg/L). The aflatoxin M 1 in the samples never exceeded the level established by the European Union., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Pre-column derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) was determined to be effective for quantitation of fumonisins B 1 and B 2 in feed. Liquid-solid extraction, clean-up using immunoaffinity solid phase extraction chromatography, and FMOC-derivatization preceded analysis by reverse phase HPLC with fluorescence. Instrument response was unchanged in the presence of matrix, indicating no need to use matrix-matched calibrants. Furthermore, high method recoveries indicated calibrants do not need to undergo clean-up to account for analyte loss. Established method features include linear instrument response from 0.04-2.5µg/mL and stable derivatized calibrants over 7days. Fortified cornmeal method recoveries from 0.1-30.0μg/g were determined for FB 1 (75.1%-109%) and FB 2 (96.0%-115.2%). Inter-assay precision ranged from 1.0%-16.7%. Method accuracy was further confirmed using certified reference material. Inter-laboratory comparison with naturally-contaminated field corn demonstrated equivalent results with conventional derivatization. These results indicate FMOC derivatization is a suitable alternative for fumonisins B 1 and B 2 quantitation in corn-based feeds., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A high-performance graphene-based micromotor strategy for simultaneous, fast, and reliable assessment of two highly concerning mycotoxins (fumonisin B1 (FB1) and ocratoxin A (OTA)) has successfully been developed. The assay principle is based on the selective recognition from aptamers to the target mycotoxins and further "on-the-move" fluorescence quenching of the free aptamer in the outer layer of unmodified reduced graphene (rGO; sensing layer) micromotors. Template-prepared rGO/platinum nanoparticles (PtNPs) tubular micromotors were synthesized rapidly and inexpensively by the direct electrodeposition within the conical pores of a polycarbonate template membrane. The new wash-free approach offers using just 1 μL of sample, a simultaneous and rapid "on-the-fly" detection (2 min) with high sensitivity (limits of detection of 7 and 0.4 ng/mL for OTA and FB1, respectively), and high selectivity. Remarkable accuracy (E r < 5%) during the mycotoxin determination in certified reference material as well as excellent quantitative recoveries (96-98%) during the analysis of food samples were also obtained. The excellent results obtained allow envisioning an exciting future for the development of novel applications of catalytic micromotors in unexplored fields such as food safety diagnosis.

A novel molecularly imprinted electrochemiluminescence (MIP-ECL) sensor based on Ru(bpy) 3 2+ -doped silica nanoparticles (Ru@SiO 2 NPs) is developed for highly sensitive detection of fumonisin B 1 (FB 1 ). Gold-nanoparticles (AuNPs), Ru@SiO 2 NPs with chitosan (CS) composites and a molecularly imprinted polymer (MIP) are assembled on a glassy carbon electrode (GCE) to fabricate an ECL platform step by step. AuNPs could greatly promote the ECL intensity and improve the analytical sensitivity according to the localized surface plasmon resonance (LSPR) and the electrochemical effect. In this surface-enhanced electrochemiluminescence (SEECL) system, AuNPs work as the LSPR source to improve the ECL intensity and Ru@SiO 2 NPs are used as ECL luminophores. In the phosphate buffer solution (PBS), the evident anodic ECL of Ru@SiO 2 on the above working electrode is observed in the presence of the template molecule fumonisin B 1 (FB 1 ), which could act as the coreactant of Ru@SiO 2 NPs due to the amino group of FB 1 . When the template molecules were eluted from the MIP, little coreactant was left, resulting in an apparent decrease of ECL signal. After the MIP-ECL sensor was incubated in FB 1 solution, the template molecules rebound to the MIP surface, leading to the enhancement of ECL signal again. On the basis of these results, a facile MIP-ECL sensor has been successfully fabricated, which exhibited a linear range from 0.001 to 100ngmL -1 with a detection limit of 0.35pgmL -1 for FB 1 . Moreover, the proposed MIP-ECL sensor displayed an excellent application in real samples., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Being able to quantify ichthyotoxic metabolites from microalgae allows for the determination of ecologically-relevant concentrations that can be simulated in laboratory experiments, as well as to investigate bioaccumulation and degradation. Here, the ichthyotoxin karmitoxin, produced by Karlodinium armiger , was quantified in laboratory-grown cultures using high-performance liquid chromatography (HPLC) coupled to electrospray ionisation high-resolution time-of-flight mass spectrometry (HRMS). Prior to the quantification of karmitoxin, a standard of karmitoxin was purified from K. armiger cultures (80 L). The standard was quantified by fluorescent derivatisation using Waters AccQ-Fluor reagent and derivatised fumonisin B₁ and fumonisin B₂ as standards, as each contain a primary amine. Various sample preparation methods for whole culture samples were assessed, including six different solid phase extraction substrates. During analysis of culture samples, MS source conditions were monitored with chloramphenicol and valinomycin as external standards over prolonged injection sequences (>12 h) and karmitoxin concentrations were determined using the response factor of a closely eluting iturin A2 internal standard. Using this method the limit of quantification was 0.11 μg·mL -1 , and the limit of detection was found to be 0.03 μg·mL -1 . Matrix effects were determined with the use of K. armiger cultures grown with 13 C-labelled bicarbonate as the primary carbon source., Competing Interests: The founding sponsors had no role in the design of the study, in the collection, analyses, or interpretation of data, in the writing of the manuscript, and in the decision to publish the results.

This study estimated exposure to multiple mycotoxins in 249 infants aged between 6 and 12 months in three agro-ecological zones of Tanzania. Maize-based complementary food intakes were estimated using two 24 h dietary recalls. Using @Risk software, probabilistic exposure assessment was conducted by modeling maize intake data (kg/kg body weight/day) with previously determined multiple mycotoxin (except for ochratoxin A (OTA) and zearalenone (ZEA), present in only a few samples) contamination data (μg/kg) in maize. Maize intakes ranged from 0.13 to 185 g/child/day (average = 59 ± 36 g/child/day). The estimated mean exposures were higher for aflatoxins (6-fold), fumonisins (3-fold), and deoxynivalenol (2-fold) than health-based guidance values of 0.017 ng/kg body weight/day, 2 μg/kg body weight/day, and 1 μg/kg body weight/day, respectively. The population at risk of exposures above the limits of health concern ranged from 12% for HT-2 toxin through 35% for deoxynivalenol to 100% for aflatoxins. The exposure varied among the agro-ecological zones. Strategies targeting multiple mycotoxins in maize are urgently needed to minimize exposures in Tanzania.

Mycotoxins are fungal secondary metabolites with bioaccumulation levels leading to their carry-over into animal fluids, organs, and tissues. As a consequence, mycotoxin determination in biological samples from humans and animals has been reported worldwide. Since most mycotoxins show toxic effects at low concentrations and considering the extremely low levels present in biological samples, the application of reliable detection methods is required. This review summarizes the information regarding the studies involving mycotoxin determination in biological samples over the last 10 years. Relevant data on extraction methodology, detection techniques, sample size, limits of detection, and quantitation are presented herein. Briefly, liquid-liquid extraction followed by LC-MS/MS determination was the most common technique. The most analyzed mycotoxin was ochratoxin A, followed by zearalenone and deoxynivalenol-including their metabolites, enniatins, fumonisins, aflatoxins, T-2 and HT-2 toxins. Moreover, the studies were classified by their purpose, mainly focused on the development of analytical methodologies, mycotoxin biomonitoring, and exposure assessment. The study of tissue distribution, bioaccumulation, carry-over, persistence and transference of mycotoxins, as well as, toxicokinetics and ADME (absorption, distribution, metabolism and excretion) were other proposed goals for biological sample analysis. Finally, an overview of risk assessment was discussed., Competing Interests: The authors declare no conflict of interest.

An epidemic fungal disease caused by Fusarium proliferatum, responsible for fumonisin production (FB1, FB2, and FB3), has been reported in the main garlic-producing countries in recent years. Fumonisins are a group of structurally related toxic metabolites produced by this pathogen. The aim of this work was to establish an enzyme-linked immunosorbent assay (ELISA) procedure, mostly applied to cereals, that is suitable for fumonisin detection in garlic and compare these results to those obtained by high-performance liquid chromatography (HPLC) and screening of fresh and dehydrated garlic for toxicological risk. The results show good correlation between the two analytical methods. In fresh symptomatic garlic, fumonisin levels were higher in the basal plates than those in the portions with necrotic spots. Among the 56 commercially dehydrated garlic samples screened, three were positive by ELISA test and only one was above the limit of quantitation. The same samples analyzed by HPLC showed the presence of FB1 in trace amounts that was below the limit of quantitation; FB2 and FB3 were absent. The results are reassuring, because no substantial contamination by fumonisins was found in commercial garlic.

A fast, easy, and cheap method for the simultaneous determination and quantification of aflatoxins (B1, B2, G1, G2), T-2 and HT-2 toxins, and fumonisins (B1, B2) in cereal-derived products was developed. This method involved a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction coupled with liquid chromatography-tandem mass spectrometry. The method validation was performed by analyzing samples spiked at four levels, and the recoveries ranged from 83.6 to 102.9%, whereas the maximum values of repeatability and within-laboratory reproducibility were 14.3 and 15.7% following the performance criteria set by the European legislation. The method was then applied for the analysis of 21 cereal-derived products purchased on the Italian market, which were correctly packaged and labeled as intended for human consumption. The co-occurrence of more than one mycotoxin in the analyzed samples could represent a risk for consumers, and the described method could be a valid alternative for their simultaneous detection in the framework of official control. Graphical Abstract ᅟ.