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Koji mold, which belongs to the Aspergillus section Nigri, is used in the production of shochu. The section Nigri is composed of very morphologically similar members that in some cases produce mycotoxins, which rises concerns as to whether the presence of mycotoxin-producing fungi in shochu producing sites can compromise consumer safety. Thus, we examined the presence of mycotoxin-producing sec. Nigri fungi in six shochu factories (named A-F) in Japan. Airborne fungal levels in the factories were determined, and a traditional koji called "kona-koji" made from the mold naturally present in factory C (Aogashima village) was analyzed. Isolates of sec. Nigri fungi were identified morphologically and confirmed via cytochrome b gene analysis. In factory A (Nago city), airborne fungal levels of sec. Nigri were 4,000 and 100 cfu/m3 in the koji-making and fermentation rooms, respectively. In factories B, C, and D, the levels were 40, >104 cfu/m3, and 100 cfu/m3, respectively. In factory F (Iki city), there were high levels of airborne white-koji mold (a white mutant of Asp. luchuensis). The most dominant fungal species of sec. Nigri was isolated and identified as Asp. luchuensis via genetic analysis. This is likely to have originated from the commercial fermentation culture used. Asp. niger and Asp. luchuensis were isolated from kona-koji. Mycotoxin production (ochratoxin and fumonisin B2) by Asp. luchuensis (eight strains) and Asp. niger (three strains) was virtually inexistent; only one strain of Asp. niger was positive for fumonisin B2. This study clearly shows that mycotoxin-producing fungi are not dominant in the fungal flora present in the shochu factories examined and therefore, that the liquor can be safely fermented.Implications: In this study, we examined the presence of mycotoxin-producing Aspergillus sec. Nigri fungi in six shochu (Japanese distilled beverage) factories. The most dominant fungal species of sec. Nigri was isolated and identified as Aspergillus luchuensis (black-koji mold). The proportion of mycotoxin-producing Aspergillus niger and Aspergillus carbonarius was very small. In addition, the Asp. niger isolated from koji mold did not have the ability to produce ochratoxins or fumonisin B. This study clearly shows that shochu can be safely fermented.

The aim of this study was to isolate Aspergillus section Nigri from onion samples bought in supermarkets and to analyze the fungal isolates by means of molecular data in order to differentiate A. niger and A. welwitschiae species from the other non-toxigenic species of black aspergilli, and detect genes involved in the biosynthesis of ochratoxin A and fumonisin B2. Aspergillus section Nigri were found in 98% (94/96) of the onion samples. Based on the results of multiplex PCR (performed on 500 randomly selected strains), 97.4% of the Aspergillus section Nigri strains were recognized as A. niger/A. welwitschiae. Around half of them were subjected to partial sequencing of the CaM gene to distinguish one from the other.Atotal of 97.9% of the isolates were identified as A. welwitschiae and only 2.1% as A. niger. The fum8 gene, involved in fumonisin B2 biosynthesis, was found in 36% of A. welwitschiae isolates, but radH and pks genes, involved in ochratoxin A biosynthesis, were found in only 2.8%. The presence/absence of fum8 gene in the A. welwitschiae genome is closely associated with ability/inability of the isolates to produce fumonisin in vitro. Based on these results,we suggest that in-depth studies are conducted to investigate the presence of fumonisins in onion bulbs. FAPESP / CAPES / CNPq

Nine domestic wine samples collected from a Japanese winery were examined for the presence of fumonisin B1 (FB1), fumonisin B2 (FB2), and fumonisin B3 (FB3), as well as ochratoxin A (OTA) and ochratoxin B (OTB). Wine samples spiked with 13C-labeled internal standards (13C34-FB1 and 13C20-OTA) were diluted with phosphate buffered saline (PBS) buffer, loaded on immunoaffinity cartridges to purify of fumonisins and ochratoxins, and subjected to liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis. The data revealed that the domestic wine samples were possibly contaminated with FB1 and FB3, in addition to FB2, whereas none of the tested wine samples were contaminated with OTA and OTB. These results suggest that Fusarium fungi can be associated with the fumonisin contamination of Japanese domestic wine, whereas Aspergillus niger seems to be frequently reported as the major causal fungus of fumonisin contamination of wine in Europe. Analysis of the intermediate samples during the wine brewing indicated that fumonisin concentrations did not increase during wine production, suggesting that fumonisin contamination did not occur during the brewing process, but was derived from the raw materials of grape berries.

Aspergillus spp. are a common contaminant of grapes and a major source of mycotoxins. China is the largest producer of grape in the world now, however, the toxigenic Aspergillus population on grape in this country is still largely unknown. In this study, a total of 345 strains were isolated from grapes of 13 main grapevine producing regions in China. Based on calmodulin gene sequences, eight species within Aspergillus section Nigri were identified. Among them, Aspergillus tubingensis (48.7%) was predominant, followed by Aspergillus welwitschiae (20.6%) and Aspergillus aculeatinus (11%). Average of contamination level was up to 64.19%, and we found the occurrence of section Nigri species on the surface of fresh grapes was significantly influenced by the climate (P2 (FB2) producers. Of those, Aspergillus carbonarius was the main OTA producer and A. welwitschiae and Aspergillus niger were the main FB2 producers with different toxigenic ability. Our results highlight the potential risk of OTA and FB2 contamination by A. carbonarius and A. welwitschiae on grape in China, respectively; management strategies should be considered for preventing and reducing the toxigenic Aspergillus and its mycotoxins.

The garlic contains sulfur bioactive compounds responsible for medicinal properties. The decrease of these compounds due to inadequate storage conditions reduces the beneficial properties and favors infection by microorganisms. Several studies have shown high frequency of garlic infected with Aspergillus section Nigri that potentially produce mycotoxin. Garlic samples were collected in markets of Brazil and a total of 32 samples (of 36) had the fungal infection with predominant genus Aspergillus (50.3%), Penicillium (34.7%), and Fusarium (11%). A total of 63% (649/1031) of infection with Aspergillus section Nigri, of which 60 isolates were selected for analysis of genetic variability that resulted in 4 clusters. Representatives of clusters were identified by the calmodulin gene. Isolates from cluster I were subdivided into A-I and identified as A. niger (16 isolates) and the isolates of clusters B-I, II, and III were identified as A. welwitschiae (43 isolates). Besides, an isolate of the IV-cluster was identified by A. luchuensis. Further, we used the multiplex PCR to verify genotypes of 59 isolates, and none of these had OTA production-associated genotype. Moreover, 19 A. welwitschiae and 15 A. niger were FB2 production-associated genotype. Our study is the first report to the incidence of garlic infection in Brazil and to show that A. welwitschiae causes most of these infections.

The fungal diseases in the vineyard reduce grape quality influencing the volatile profile and consequently impair the aroma, taste, and color of juice and wine. In addition to this sensory effect, the occurrence of filamentous fungi in grapes may result in mycotoxins, including alternariol, aflatoxins, tenuazonic acid, fumonisin B2, ochratoxin A (OTA), patulin citrinin, among others. OTA is the most studied mycotoxin in grape products and can effectively represent a concern for industries. This short review discloses that promising results have been obtained with the use of antagonistic microorganisms or natural products for the toxigenic fungi control in the vineyard. Reduction of OTA levels throughout the production stages of juice and wine is well documented in the literature. However, studies taking into account the impacts of climate change on vine and the resilience of toxigenic fungi are still a challenge. In addition, the co-occurrence of OTA (including modified forms) and other mycotoxins in grapes products of different cultivars and regions needs to be evaluated to verify the risk of human exposure to these compounds.

This study was conducted to detect and quantify three mycotoxins Dioxynivalenol, Nivalenol and Fumonisin B2 in seeds of seven wheat cultivars planted in 17 wheat fields in Basra province, Iraq. This was done by using High Performance Liquid chromatographs analysis. The results revealed that Fumonisin B2 was the predominant mycotoxin, which present in 10 fields. The lowest concentration rate of this mycotoxin was 110 µg/Kg and the maximum was 11,228 µg/Kg. Dioxynivalenol as a trichothecene was in the second level detected in 6 fields with a minimum concentration of 8 µg/Kg and a maximum of 1,060 µg/Kg. Nivalenol was found only in 4 fields ranging from 272-1900 µg/Kg. Fumonisin B2 Only three fields showed co-occurrence of two mycotoxins (Fumonisin B2 and Nivalenol) in each, but with various concentration rates. The seven cultivars tested in this study were varied in their reactions to subjected mycotoxins. Adana 99 (A. 99) cultivar showed the highest concentration rate of both Fumonisin B2 and Nivalenol, which present with average percentage of 64% and 58% respectively. While, for Dioxynivalenol, Ebaa 99 (E.99) was on the top, it occurred in 6 fields ranging from 8-1060 µg/Kg with an average percentage of 43%.

Our study investigated the occurrence of a wide range of mycotoxins in coffee, most of them rarely analyzed in this commodity, and the safety relevance of mycotoxin exposure from coffee consumption. A new liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the analysis of 31 regulated and emerging mycotoxins was therefore developed and validated. Quantification was performed by the matrix-matched calibration curve approach. Analyses were conducted on green coffee which permitted to reach low limits of quantification, i.e. from 0.25 μg/kg to 150 μg/kg. A total of 71 green coffee beans samples, harvested in 2015 and 2016, were collected from various geographical origins (nine countries) and comprised of Arabica or Robusta species. Types of post-harvest processes and storage times in bags were also considered during sample collection. Diversity, occurrence and levels of mycotoxins in the surveyed green coffee samples were quite low. The mycotoxins most frequently found and at highest levels were Ochratoxin A, Fumonisin B2, Sterigmatocystin, Beauvericin and Enniatin A with maximum levels at 12.2, 76.3, 19.8, 4.4 and 1.7 μg/kg, respectively. Contamination with Aflatoxin B1 was sporadic with maximum level at 1.2 μg/kg. Co-occurrence of multiple mycotoxins per sample was rare, at very low levels and mostly limited to two different toxins. A risk assessment performed on our data set indicates that coffee is a minor contributor to mycotoxin exposure from the diet and is not expected to represent a safety concern.

Ülkemizde mısır üretimindeki artış bir yandan sürerken, bir yandan da dünyada mısırın soyadan sonra en çok genetiği değiştirilmiş ürün olması nedeniyle tüketicinin mısır ile ilgili algısı her geçen gün daha olumsuz olmaktadır. Genetiği değiştirilmiş mısırın potansiyel olumsuz etkileri tartışılmakta iken, mısırda insan sağlığı için gözden kaçmaması gereken en büyük tehdit mikotoksinlerdir. Bu çalışmada, Türkiye‘nin 24 ilinde bulunan 552 köyün 634 tarlasından toplanan mısır numuneleri GDO, aflatoksin B1, toplam aflatoksin, fumonisin B1, fumonisin B2, T-2 toksin, HT-2 toksin, zearalenon ve deoksinivalenol yönünden incelenmiştir. Tarama yapılan mısır numunelerinin hiçbirinde transgenik elemente rastlanmamıştır. Analiz edilen 634 numunenin sadece bir tanesinde Türk Gıda Kodeksi limitinin üzerinde aflatoksin B1 miktarına rastlanmış, numunelerin toplam aflatoksin değerleri Türk Gıda Kodeksi limitinin altında çıkmıştır. Paçal oluşturulan numunelerin hiçbirinde T-2 toksin, HT-2 toksin, zearalenon ve deoksinivalenol tespit edilmezken, fumonisin tespit edilen numunelerin toplam fumonisin miktarı Türk Gıda Kodeksi limitinin altında bulunmuştur. Bu sonuçlar, yerli üretim mısırların gıda güvenliği açısından uygun standartları sağladığını göstermektedir.

Fusarium verticillioides is a prolific producer of useful secondary metabolites such as naphthoquinone pigments, monoterpenes, and sesquiterpenes, as well as the harmful mycotoxins fumonisins. A strategy to increase their production includes creating a proper nutritional environment that enables the fungus to produce the compounds of interest. The aim of the present work was to study the effect of different carbon sources (glucose, fructose, xylose, sucrose, and lactose) on secondary metabolites biosynthesis in F. verticillioides submerged cultures. The production of volatile terpenes was evaluated through gas chromatography coupled to mass spectrometry. The quantification and identification of pigments was conducted using a UV/VIS spectrophotometer and NMR spectrometer, respectively. The quantification of fumonisin B1 and fumonisin B2 was performed by high-performance liquid chromatography. Our results showed that the biosynthesis of naphthoquinone pigments, monoterpenes, and sesquiterpenes was highest in cultures with fructose (13.00 ± 0.71 mmol/g), lactose [564.52 × 10−11 ± 11.50 × 10−11 μg/g dry weight (DW)], and xylose (54.41 × 10−11 ± 1.55 × 10−11 μg/g DW), respectively, with fumonisin being absent or present in trace amounts in the presence of these carbon sources. The highest biosynthesis of fumonisins occurred in sucrose-containing medium (fumonisin B1: 7.85 × 103 ± 0.25 × 103 μg/g DW and fumonisin B2: 0.38 × 103 ± 0.03 × 103 μg/g DW). These results are encouraging since we were able to enhance the production of useful fungal metabolites without co-production with harmful mycotoxins by controlling the carbon source provided in the culture medium.

Along with Aspergillus and Penicillium spp., mycotoxins produced by Fusarium species are important contaminants in food and feed industry. This study was designed to investigate the biodegradation potential of a collection of Bacillus spp. In different substrates artificially contaminated with single or multiple Fusarium mycotoxins. B. megaterium BM344-1 and B. pumilus BP344-3 showed the highest degradation (100%) of zearalenone (ZEN) on liquid Luria Bertani (LB) medium. Bacillus cereus BC344-2, Bacillus cereus BC399-2, Bacillus simplex BS350-3 and Paenibacillus favisporus PF341, showed significant biodegrading capacity on T-2 toxin, leading to 72, 70, 71 and 88% decontamination, respectively. Complete degradation (100%) of ZEN was observed at a contamination level of 0.375 μg/ml in LB, while showing a decreasing trend of degradation (87 and 84%) at increasing levels of mycotoxins (0.750 and 1.50 μg/ml LB medium, respectively). Likewise, BM344-1 showed a decreasing degradation trend (81, 78% and 55%) at increasing T-2 contamination levels (0.3, 0.6 and 1.2 μg/ml, respectively). The ability of BM344-1 to degrade fumonisin B1 (FB1) and fumonisin B2 (FB2) was 14 and 12%, respectively. In LB medium concurrently contaminated with multiple mycotoxins (ZEN + T-2+HT-2+FB1+FB2), the degradation ability of BM344-1 was not affected by low-to-medium levels of each mycotoxin, while it was significantly lowered upon addition of each toxin at higher levels. In vivo mycotoxin degradation potential of BM344-1 tested on artificially contaminated maize showed 25–44% reduction of ZEN and 15–19% reduction of T-2, with a non-significant effect in case of combined mycotoxins contamination. B. megaterium BM344-1, having significant biodegrading properties on multiple Fusarium mycotoxins, can be considered for its potential application as a mycotoxin decontamination agent in the food industry.

In this work, a retrospective screening based on ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) based on Orbitrap-Q-Exactive Focus™ was used to check the occurrence of regulated and emerging mycotoxins in bulk milk samples. Milk samples were collected from dairy farms in which corn silage was the main ingredient of the feeding system. The 45 bulk milk samples were previously analyzed for a detailed untargeted metabolomic profiling and classified into five clusters according to the corn silage contamination profile, namely: (1) low levels of Aspergillus- and Penicillium-mycotoxins, (2) low levels of fumonisins and other Fusarium-mycotoxins, (3) high levels of Aspergillus-mycotoxins, (4) high levels of non-regulated Fusarium-mycotoxins, (5) high levels of fumonisins and their metabolites. Multivariate statistics based on both unsupervised and supervised analyses were used to evaluate the significant fold-change variations of the main groups of mycotoxins detected when comparing milk samples from clusters 3, 4, and 5 (high contamination levels of the corn silages) with cluster 1 and 2 (low contamination levels of the corn silages). Overall, 14 compounds showed a significant prediction ability, with antibiotic Y (VIP score = 2.579), bikaverin (VIP score = 1.975) and fumonisin B2 (VIP score = 1.846) being the best markers. The k-means clustering combined with supervised statistics showed two discriminant groups of milk samples, thus revealing a hierarchically higher impact of the whole feeding system (rather than the only corn silages) together with other factors of variability on the final mycotoxin contamination profile. Among the discriminant metabolites we found some Fusarium mycotoxins, together with the tetrapeptide tentoxin (an Alternaria toxin), the α-zearalenol (a catabolite of zearalenone), mycophenolic acid and apicidin. These preliminary findings provide new insights into the potential role of UHPLC-HRMS to evaluate the contamination profile and the safety of raw milk to produce hard cheese.

Mycotoxin-producing Aspergilli (Circumdati, Flavi, and Nigri), usually associated with contaminated food, may also cause respiratory disorders and are insufficiently studied in water-damaged indoor environments. Airborne (N = 71) and dust borne (N = 76) Aspergilli collected at post-flood and control locations in Croatia resulted in eleven different species based on their calmodulin marker: A. ochraceus, A. ostianus, A. pallidofulvus, A. sclerotiorum, and A. westerdijkiae (Circumdati), A. flavus (Flavi), and A. tubingensis, A. welwitschiae, A. niger, A. piperis, and A. uvarum (Nigri). Most of the airborne (73%) and dust borne (54%) isolates were found at post-flood locations, and the highest concentrations measured in indoor air (5720 colony-forming units (CFU)/m3) and dust (2.5 &times, 105 CFU/g) were up to twenty times higher than in the control locations. A. flavus dominated among airborne isolates (25%) at the unrepaired locations, while 56% of the dust borne Aspergilli were identified as A. tubingensis and A. welwitschiae. The ability of identified isolates to produce mycotoxins aflatoxin B1 (AFB1), fumonisin B2 (FB2), and ochratoxin A were assessed by LC-MS analysis. All ochratoxin A (OTA)-producing Circumdati belonged to A. westerdijkiae (13.7 &plusmn, 15.81 &micro, g/mL), in the section, FlaviA. flavus produced AFB1 (2.51 &plusmn, 5.31 &micro, g/mL), while A. welwitschiae and A. niger (section Nigri) produced FB2 (6.76 &plusmn, 13.51 &micro, g/mL and 11.24 &plusmn, 18.30 &micro, g/mL, respectively). Water damage dominantly supported the occurrence of aflatoxigenic A. flavus in indoor environments. Yet unresolved, the causal relationship of exposure to indoor Aspergilli and adverse health effects may support the significance of this research.

Coix seed is an important food and traditional Chinese medicine in China and other Asian countries. Notably, coix seed is currently being used as a traditional medicine for the treatment of COVID-19 in China. However, coix seeds are generally contaminated by mycotoxins, and this risk cannot be ignored. In this paper, we developed a method that involves direct extraction and UHPLC-HRMS analysis for the simultaneous detection of 24 mycotoxins in coix seeds. UHPLC-HRMS instrument and data acquisition parameters, and the sample pretreatment were optimised. One-step extraction showed several advantages compared to the three commercial solid-phase extraction clean-up methods, including ease of use, reduced time of sample preparation, low cost, good recovery, and acceptable matrix effect. The method validation results indicate that all mycotoxins have good linearity and sensitivity. Recoveries were between 74.2-101.1%, and RSD ranged from 0.1-5.8%. The LOQs for 24 mycotoxins were in the range of 0.5-100 µg/kg. To survey the contamination levels of these mycotoxins in commercial coix seeds, more than 70 samples were collected from Chinese markets and were analysed using the newly developed method. Zearalenone (positive ratio: 98.7%, range:1.1-1562 µg/kg), deoxynivalenol (positive ratio: 87%, range: 8.4-382.5 µg/kg), nivalenol (positive ratio: 85.7%, range: 26.8-828.2 µg/kg), fumonisin B1 (positive ratio: 84.4%, range:2.5-314.5 µg/kg), fumonisin B2 (positive ratio: 75.3%, range:1.6-72.8 µg/kg), fumonisin B3 (positive ratio: 48%, range:1.0-203.6 µg/kg), aflatoxin B1 (positive ratio: 29.9%, range: 0.39-14.7 µg/kg), sterigmatocystin (positive ratio: 29.9%, range: 1.4-51.6 µg/kg), and tenuazonic acid (positive ratio: 19.5%, range 36.1-105.7 µg/kg) were the most frequent mycotoxin contaminants. These results highlight the importance of routine monitoring and control of mycotoxins in coix seeds.

The occurrence of diseases in the cultivation of maize (Zea mays L.) leads to the loss of grain yield and the simultaneous deterioration of its quality. Fungi of the genus Fusarium spp. pose the greatest threat to maize cultivation. These fungi occur at all stages of the plant&rsquo, s development, causing Fusarium seedling blight, root rot, foot rot, and ear rot, including grains. Therefore, the aim of the conducted field studies was to determine the influence of selected agriculture factors on mycotoxin contents in maize grain and ear core. Moreover, it should be noted that the presence of mycotoxins in food products in terms of legal regulations is quite a fundamental barrier both in domestic and international trade. The field experiment was carried out at the Department of Agronomy of Poznań University of Life Sciences, on the fields of the Experimental and Educational Unit in Swadzim, Poland, in 2013&ndash, 2014, in four field replicates. The study involved the following factors: soil sowing preparation method, types of variates, and fertilization method, all of which shape mycotoxin accumulation. The results indicated that the main effects of years were significant for all observed traits for both the core and grain. The concentrations of deoxynivalenol, nivalenol, and zearalenone were higher in 2014 than in 2013 for both core and grain. The concentrations of fumonisin B1 and fumonisin B2 were higher in the first year of study for core and grain. The direct sowing of maize significantly affects plants&rsquo, health status, expressed by an increase in mycotoxin accumulation. Traditional hybrid SY Cooky characterized higher values of observed traits than &ldquo, stay-green&rdquo, hybrid Drim (except fumonisin B2 for cob). The positive effect of the row method of NP fertilizer application is based on a decrease in mycotoxin content.

Climate change is primarily manifested by elevated temperature and carbon dioxide (CO2) levels and is projected to provide suitable cultivation grounds for pests and pathogens in the otherwise unsuitable regions. The impacts of climate change have been predicted in many parts of the world, which could threaten global food safety and food security. The aim of the present work was therefore to examine the interacting effects of water activity (aw) (0.92, 0.95, 0.98 aw), CO2 (400, 800, 1200 ppm) and temperature (30, 35 &deg, C and 30, 33 &deg, C for Fusarium verticillioides and F. graminearum, respectively) on fungal growth and mycotoxin production of acclimatised isolates of F. verticillioides and F. graminearum isolated from maize. To determine fungal growth, the colony diameters were measured on days 1, 3, 5, and 7. The mycotoxins produced were quantified using a quadrupole-time-of-flight mass spectrometer (QTOF-MS) combined with ultra-high-performance liquid chromatography (UHPLC) system. For F. verticillioides, the optimum conditions for growth of fumonisin B1 (FB1), and fumonisin B2 (FB2) were 30 &deg, C + 0.98 aw + 400 ppm CO2. These conditions were also optimum for F. graminearum growth, and zearalenone (ZEA) and deoxynivalenol (DON) production. Since 30 &deg, C and 400 ppm CO2 were the baseline treatments, it was hence concluded that the elevated temperature and CO2 levels tested did not seem to significantly impact fungal growth and mycotoxin production of acclimatised Fusarium isolates. To the best of our knowledge thus far, the present work described for the first time the effects of simulated climate change conditions on fungal growth and mycotoxin production of acclimatised isolates of F. verticillioides and F. graminearum.

Fumonisins contamination of food commodities is a worldwide problem, especially for maize. The ability to produce fumonisinsis a trait of several species of Fusarium, mainly F. verticillioides and F. proliferatum on maize, and some Aspergillus species.A. nigerand its sister speciesA. welwitschiae, can contribute to fumonisin B2(FB2) accumulation in maize kernels, although to a lesser extent than fumonisin-producingFusariumspecies. Fumonisins risk monitoring represents an effective strategy in the integrated approach for mycotoxin risk management and reduction. The availability of a user-friendlymolecular assay for the detection oftoxigenic fungal species represents a valuable tool in understanding and managing upcoming mycotoxin contamination. In this study, we developed a LAMP assay, based on the detection offum10, for a rapid and specific molecular detection of FB2-producingA. nigerandA. welwistchiae, potentially useful to perform monitoring directly “on site” in maize chain. Results showed that very low amounts of conidia are suitable to detect the presence of the target gene, thus providing information about the presence of FB2-producingAspergillusspecies and the possible upcoming fumonisins contamination in maize. The assay was combined with a suitable protocol for “in field”crude DNA extraction and a colorimetric method for easy naked-eye evaluationof results, offering a reliable and user-friendly tool to support effective reduction strategies of mycotoxin contamination in crop management programs.

Investigating the in vitro fumonisin biosynthesis and the genetic structure of Fusarium verticillioides populations can provide important insights into the relationships between strains originating from various world regions. In this study, 90 F. verticillioides strains isolated from maize in five Mediterranean countries (Italy, Spain, Tunisia, Egypt and Iran) were analyzed to investigate their ability to in vitro biosynthesize fumonisin B1, fumonisin B2 and fumonisin B3 and to characterize their genetic profile. In general, 80% of the analyzed strains were able to biosynthesize fumonisins (range 0.03&ndash, 69.84 &mu, g/g). Populations from Italy, Spain, Tunisia and Iran showed a similar percentage of fumonisin producing strains (&gt, 90%), conversely, the Egyptian population showed a lower level of producing strains (46%). Significant differences in fumonisin biosynthesis were detected among strains isolated in the same country and among strains isolated from different countries. A portion of the divergent FUM1 gene and of intergenic regions FUM6-FUM7 and FUM7-FUM8 were sequenced to evaluate strain diversity among populations. A high level of genetic uniformity inside the populations analyzed was detected. Apparently, neither geographical origin nor fumonisin production ability were correlated to the genetic diversity of the strain set. However, four strains from Egypt differed from the remaining strains.

Fumonisins (FB1+FB2) and deoxynivalenol (DON) are mycotoxins produced by Fusarium species that might be present in maize and maize products. Knowledge on their occurrence in nixtamalized maize from Mexico together with an accompanying risk assessment are scarce, while nixtamalized maize is an important food in Mexico. This study presents the occurrence of FB1 + FB2 and DON in nixtamalized maize samples collected in Mexico City and analyses their distribution and resulting estimated daily intake for Mexican consumers by a probabilistic approach using a two-dimensional Monte-Carlo simulation. The results obtained reveal that for FB1 + FB2, 47% of the Mexican men and 30% of the Mexican women might exceed the provisional tolerable daily intake (PMTDI) of 2 &micro, g/kg bw/day for fumonisins and for DON, 9% of men and 5% of women would be exceeding the PMTDI of 1 &micro, g/kg bw/day, corresponding to the high consumers. The results raise a flag for risk managers in Mexico, to consider regulations and interventions that lower mycotoxin levels in nixtamalized maize for human consumption.

Some species from Aspergillus section Nigri are morphologically very similar and altogether have been called A. niger aggregate. Although the species included in this group are morphologically very similar, they differ in their ability to produce mycotoxins and other metabolites and their taxonomical status has evolved continuously. Among them, A. niger and A. welwitschiae are ochratoxin A and fumonisin B2 producers and their detection and/or identification is of crucial importance for food safety. The aim of this study was the development of a real-time PCR-based method for simultaneous discrimination of A. niger and A. welwitschiae from other species of the A. niger aggregate isolated from coffee beans. One primer pair and a hybridization probe specific for detection of A. niger and A. welwitschiae strains were designed based on the BenA gene sequences, and used in a Real-time PCR assay for the rapid discrimination between both these species from all others of the A. niger aggregate. The Real-time PCR assay was shown to be 100% efficient in discriminating the 73 isolates of A. niger/A. welwitschiae from the other A. niger aggregate species analyzed as a negative control. This result testifies to the use of this technique as a good tool in the rapid detection of these important toxigenic species.

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 B2was observed in 2 (7%) samples with contamination levels ranging from 4.99 to 16.2 μg/kg. The simultaneous detection of fumonisin B2in the same contaminated samples with ochratoxins indicates the fungal attack by Aspergillus nigerspecies during storage. Only 1 sample was contaminated with aflatoxin B1(14.4 μg/kg) and B2(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 B2and ochratoxin A and B in addition to a wide range of other fungal metabolites in date palm fruits. 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.

Fumonisins are well known as mycotoxins produced by various Fusarium species. Recently Aspergillus niger has been reported to be a fumonisin B2 (FB2) producer. Aspergillus niger is a member of Aspergillus section Nigri. Members of this section are common food contaminants and are also distributed widely in the environment. This study aimed to determine 1) optimum culture conditions of A. niger for fumonisin production including growth medium, temperature and incubation period and 2) fumonisin production among isolates of Aspergillus section Nigri and closely related species isolated from Japanese food and environmental samples. Growth on Czapek yeast extract broth +5% NaCl (CYBS) at 28°C for 7 days resulted in the highest levels of FB2 production as determined by quantitative LC-MS/MS of culture extracts. Sixty-two isolates were collected from various foods in domestic markets as well as from soil and air. The isolates principally separated into two groups; A. niger and A. luchuensis/A. piperis/A. tubingensis, following molecular phylogenetic analysis. ELISA using the tip culture method was shown to be suitable for screening of the fumonisin-producing strains. Phylogenic analysis of Aspergillus section Nigri isolates from food and environmental samples indicated that fumonisin producing strains could be grouped into the A. niger clade. Nineteen of 35 (54%) isolates classified as A. niger were FB2 producers. The current study suggests that FB2-producing A. niger are distributed throughout several regions of Japan.

Spain is a relevant producer of oats (Avena sativa), but to date there has been no study on the occurrence/co-occurrence of mycotoxins in oats marketed in Spain. The present study is addressed to overcome this lack of knowledge. One hundred oat kernel samples were acquired across different Spanish geographic regions during the years 2015–2019 and analyzed for mycotoxin content using an ultra-high performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC–ESI–MS/MS) method and matrix-matched calibration. The focus was on the regulated mycotoxins although other relevant mycotoxins were considered. The percentage of incidence (levels ≥ limit of detection), mean and range (ng/g) of mycotoxins were as follows: zearalenone (66%, mean 39.1, range 28.1–153), HT-2 toxin (47%, mean 37.1, range 4.98–439), deoxynivalenol, (34%, mean 81.4, range 19.1–736), fumonisin B1 (29%, mean 157.5, range 63.2–217.4), and T-2 toxin, (24%, mean 49.9, range 12.3–321). Fumonisin B2, 3-acetyldeoxynivalenol, aflatoxins B1, B2, and G2, and ochratoxin A were also detected at low levels, but aflatoxin G1 was not. The maximum limits established by the European Commission for unprocessed oats were not exceeded, except for zearalenone (in one sample), and the sum of aflatoxins (in two samples). Mycotoxin co-occurrence at quantifiable levels in the same sample (two to five combinations) was found in 31% of samples. The most common mixtures were those of HT-2 + T-2 toxins alone or together with deoxynivalenol and/or zearalenone.

Maize, which contributes to a large portion of the African diet and serves as the base substrate for many fermented cereal products, has been reported to be contaminated with fumonisins. This study aimed to evaluate the in vitro ability of predominant lactic acid bacteria (LAB) in African traditional fermented maize based foods (ogi and mahewu) to bind fumonisin B1 (FB1) and B2 (FB2), as well as the stability of the complex at different pH and temperatures, in particular observed during ogi fermentation and under its storage conditions (time, temperature). The percentage of bound fumonisins was calculated after analysing the level of fumonisins not bound to LAB after a certain incubation time, by HPLC. The results revealed the ability of all tested LAB strains to bind both fumonisins, with binding efficiencies varying between strains and higher for FB2. Binding of fumonisins increased with a decrease in pH from 6 to 4 (observed during the ogi fermentation process) and from 4 to 2 (acidic pH in the stomach), and an increase in temperature (from 30 to 37 °C). The percentage of FB1 and FB2 bound to LAB at pH 4 decreased after 6 days of storage at 30 °C for all LAB strains, except for Lactobacillus plantarum (R1096) for which it increased. Lactobacillus species (L. plantarum and Lactobacillus delbrueckii) were the most efficient in binding FB1 and FB2, whereas Pediococcus sp. was less efficient. Therefore, the Lactobacillus strains tested in this study can be recommended as potential starter cultures for African traditional fermented maize based foods having detoxifying and probiotic properties.

Mycotoxins pose a serious worldwide threat to the safety of numerous food commodities. Red wine is prone to contamination from ochratoxin A, produced by black-spored Aspergillus spp.; it was recently discovered that some of these species can also produce the mycotoxin fumonisin B2. Although wine surveys in most world regions have determined that mean ochratoxin A levels are below the European Union’s recommended guideline maximum (2 μg/L), monitoring toxin levels in poor vintages and in emerging regions is imperative to ensure safety. The hot, humid environment of the southeastern United States proves challenging to winegrapes because of opportunistic fungal growth and high disease pressure, yet 550+ wineries exist in this expanding region. To determine the mycotoxin potential in southeastern vineyards, 30 cluster samples (representing 10 grape varieties, eight vineyards, and three states) were collected during the 2013 harvest and analyzed for ochratoxin A and fumonisins. Additionally, nearly 200 bottles of 100% southeastern U.S. red vinifera wines were tested for mycotoxins via liquid chromatography–tandem mass spectrometry (LC-MS/MS). These wines represent 18 grape varieties grown across six states during vintages between 2001 and 2013. Only four out of 157 (2.5%) wine samples tested positive for ochratoxin A, with the highest toxin quantity being at a safe level of 0.48 μg/L. Most wines (25/27, 92.6%) contained total fumonisins (B1, B2, and B3) at extremely low concentrations (

Two Fusarium species were identified in mouldy maize ears with the highest frequency during 2005–2014 in 7 seasons: F. subglutinans (3.1–42.0%) and F. verticillioides (44.1–70.3%). Two other species were also found but with lower frequency: F. graminearum (1.0–13.0%) and F. poae (1–45.7%). In 2005 fumonisin FB1, and in 2013 and 2014 three fumonisins (FBs) – FB1, FB2 and FB3 — were identified in harvest samples. The Fusarium-damaged kernel (FDK) fraction contained almost the totality of mycotoxins (90.0–95.0%), while healthy looking kernels (HLK) contained only below 5.0 to 10.0%. Kernels naturally infected by F. verticillioides and F. proliferatum contained (in mg kg−1) up to 710.00 of fumonisin B1, up to 209.72 of fumonisin B2 and up to 35.72 of fumonisin B3.

Several studies reported adverse respiratory health effects in workers exposed to ambient contaminants in bakeries. The aim of this study was to examine worker exposure to fungi and mycotoxins in Portuguese bakeries in order to develop new policies in occupational health. Environmental samples such as air, surfaces, settled dust and electrostatic dust collector (EDC) were collected in 13 bakeries for fungal and mycotoxins assessment. Air samples obtained by impaction were performed applying malt extract agar (MEA) supplemented with chloramphenicol (0.05%) and dichloran glycerol (DG18) agar-based media. Air samples collected through impinger method were determined as well for fungal detection by molecular tools of Aspergillus sections and mycotoxins. The highest median value for fungal load was 1053 CFU&middot, m&minus, 3 and 65.3% (32 out of 49) of the sampling sites displayed higher fungal load than limits imposed by the World Health Organization. Aspergillus genera was found in air, surface swabs and EDC. Molecular tools were effective in measuring Aspergillus section Fumigati in 22.4% on air, 27.8% on surface swabs and in 7.4% in EDC and Aspergillus section Versicolores in one air sample. All settled dust samples showed contamination with six to eight mycotoxins in each sample. The mycotoxins detected were deoxynivalenol-3-glucoside, deoxynivalenol, zearalenone, 15-acetyldeoxynivalenol, monoacetoxyscirpenol, diacetoxyscirpenol, fumonisin B1, fumonisin B2, griseofulvin, HT2, ochratoxin A, ochratoxin B and mycophenolic acid. Industrial hygienists and exposure assessors should rely on different sampling methods (active and passive) and different assays (culture based and molecular methods) to obtain an accurate risk characterization regarding fungal burden (fungi and mycotoxins). Additionally, the awareness for the raw material as a potential mycotoxins indoor contamination source is important.

A total of 105 different types of poultry feed samples from South Africa were simultaneously analysed for the presence of 16 mycotoxins using ultra-high-performance liquid chromatography coupled to a triple quadrupole mass spectrometer (UHPLC-MS/MS). The data revealed the presence of 16 mycotoxins in the various poultry feed samples. Fumonisin B1 (FB1) was the most dominant recovered from 100% of samples analysed at concentrations ranging between 38.7 and 7125.3 &micro, g/kg. This was followed by zearalenone (ZEN) (range: 0.1&ndash, 429 &micro, g/kg) and deoxynivalenol (DON) (range: 2.5&ndash, 154 &micro, g/kg). Samples were also found to be contaminated with fumonisin B2 (FB2) (range: 0.7&ndash, 125.1 &micro, g/kg), fumonisin B3 (FB3) (range: 0.1&ndash, g/kg), &alpha, zearalenol (&alpha, ZEL) (range: 0.6&ndash, 20 &micro, g/kg ), &beta, zearalenol (&beta, ZEL) (range: 0.2&ndash, 22.1 &micro, g/kg), 3-acetyldeoxynivalenol (3-ADON) (range: 0.1&ndash, 12.9 &micro, g/kg) and 15-acetyldeoxynivalenol (15-ADON) (range: 1.7&ndash, 41.9 &micro, g/kg). Alternaria mycotoxin, i.e., Alternariol monomethyl ether (AME) was recovered in 100% of samples at concentrations that ranged from 0.3&ndash, 155.5 &micro, g/kg. Aflatoxins (AFs) had an incidence rate of 92% with generally low concentration levels ranging from 0.1&ndash, 3.7 &micro, g/kg. Apart from these metabolites, 2 type A trichothecenes (THs), i.e., HT-2 toxin (HT-2) (range: 0.2&ndash, 5.9 &micro, g/kg) and T-2 toxin (T-2) (range: 0.1&ndash, 15.3 &micro, g/kg) were also detected. Mycotoxin contamination in South African poultry feed constitutes a concern as correspondingly high contamination levels, such as those observed herein are likely to affect birds, which can be accompanied by severe health implications, thus compromising animal productivity in the country. Such exposures, primarily to more than one mycotoxin concurrently, may elicit noticeable synergistic and or additive effects on poultry birds.

Twenty samples of rough rice (Oryza sativa) (unpolished kernels) collected during the 1995 harvest season from Arkansas (seven samples) and Texas (13 samples) were obtained from rice fields known to include plants with symptoms of Fusarium sheath rot putatively caused by Fusarium proliferatum. Samples were analyzed for fumonisin B1 (FB1) at three laboratories using three different extracting solvents by high-performance liquid chromatography (HPLC) or enzyme-linked immunosorbent assay (ELISA) methods. Forty percent of the samples were positive for FB1 at levels ≤4.3 μg/g by HPLC. The same samples contained FB1 at ≤3.6 μg/g when measured by an ELISA method. Most samples that were positive for FB1 were positive for fumonisin B2 (FB2) and fumonisin B3 (FB3) by HPLC at levels ≤1.2 μg/g. Very good agreement was obtained among the two laboratories using HPLC methods and the third using ELISA. Shelling of the unpolished rice results in hull and brown rice fractions. In a sample that contained 4.3 μg/g in whole kernels, the fumonisin level was very high in hulls (≤16.8 μg/g) and low in brown rice (≤0.9 μg/g). Milling of brown rice results in bran and white rice fractions. Fumonisins were found in bran at a level of ≤3.7 μg/g but were below the level of detection by HPLC in white rice. The presence of fumonisins (FB1, FB2, and FB3) was confirmed by fast atom bombardment/mass spectrometry. This is the first report of fumonisins in naturally contaminated rice in the United States.

International audience; Dried leaves and stems of Ilex paraguariensis St. Hil. (yerba mate) are used to make a popular beverage in some countries of South America, commonly known as “chimarrão”. The present study was designed to evaluate the occurrence of toxigenic Aspergillus in yerba mate in order to define the mycotoxin risk associated with this foodstuff. All samples tested were positive for fungal contamination, and the fungal load per sample ranged from 2.0 × 102 to 1.6 × 104 CFU/g. Aspergillus section Nigri was found in all samples and represented 76.5% of the total fungi isolated. Aspergillus section Circumdati, Aspergillus section Flavi and Aspergillus section Cremei were found at low frequencies. Thirteen different Aspergillus species were identified. The most common species found was A. luchuensis, which does not produce any harmful toxin for humans. A. niger, A. welwitschiae, A. flavus and A. novoparasiticus, all potentially toxigenic species, were found only in small quantities. The A. niger and A. welwitschiae strains were cultured to test for ochratoxin A and fumonisin B2 production. Only one strain producing ochratoxin A was found, but approximately 29% of the strains were positive for fumonisin B2. The A. flavus and A. novoparasiticus strains were tested for aflatoxins production, and 63% were positive. A. pallidofulvus, recently assigned to A. section Circumdati, was reported for the first time in herbs. All A. pallidofulvus strains analyzed in this study were negative for ochratoxin A production. In conclusion, A. section Nigri occurs with high frequency in yerba mate, and A. luchuensis is the predominant species. Although toxigenic species were found in this herb, the incidence was low.

The occurrence of black aspergilli in onions has been reported as frequent, and this group of fungi harbors potentially toxigenic species. In addition, Aspergillus niger has been reported as the causative agent of black mold rot, an important postharvest disease that causes damage throughout the world. Brazil stands out as one of the world’s largest onion producers. However, few studies have been conducted to investigate the mycobiota in Brazilian onions. For this reason, we investigated the mycobiota of 48 market (n = 25) and field (n = 23) onion bulb samples. Nineteen soil samples were collected from the same fields and evaluated. In field onions and soil samples, Penicillium spp. was the prevalent fungal group, whereas in market samples A. section Nigri was the most frequent group. Due to the taxonomic complexity of this group, species identification was supported by phylogenetic data (CaM gene). A. welwitschiae was the most prevalent species in market samples. Black aspergillus strains were evaluated for fumonisin B2 (FB2) and ochratoxin A (OTA) production. Overall, 53% and 2.2% of the strains produced FB2 and OTA, respectively. The occurrence of FB2 and OTA was also investigated in onion bulb samples but none showed contamination with these mycotoxins. Fapesp / Capes / CNPq

Plantain is a popular dietary staple in Central and West African countries due to its versatility and excellent nutritional value. Mature unripe plantain finger is usually processed into dried chips by processors using open sun-drying method to reduce the moisture content and extends its shelf life, but without controlling the unit operations that affect product quality. Thus, this study aimed at assessing the mycotoxin profile of solar tent-dried and open sun-dried plantain chips in Nigeria. Fifty (50) dried plantain chips [10 samples produced from the solar tent-dried, 10 samples from open sun dried, and 30 samples from local processors (9 from Akure South and 21 from Idanre Local Governments)] were analyzed for constituent mycotoxins using Liquid Chromatography Tandem-Mass Spectrometry (LC-MS/MS). The result reveals that all the regulated mycotoxins (Aflatoxin B1, B2, G1, G2, Ochratoxin A, Fumonisin B1, Fumonisin B2, Zearalenone, T-2 Toxin, HT-2 Toxin and, Deoxynivalenol) were below the detectable limits in the dried plantain chips. Aflatoxin B1 and G1, which were considered as the most regulated mycotoxin, were below the limits of detection (0.16–0.22 μg/kg) in all the samples. Only 23 analytes were detected at concentrations higher than their respective limits of detection (LOD) in 2% or more of the 50 dried plantain chips investigated, with reference to the prevalence of the non-regulated mycotoxins. Thus, all regulated mycotoxins produced by Aspergillus, Penicillium and Fusarium as stipulated by the Commission of the European Union were found at concentrations which are toxicologically acceptable in many other crops, particularly in the solar tent-dried plantain chips compared to those from the open sun-dried and local processors. Therefore, the use of a solar tent in drying plantain chips and other agricultural products is encouraged for the safety of human consumption. The outcome of this study provides useful information regarding the possible safety of plantain chips in Nigeria.

A sensitive and high-throughput method was established and validated for the simultaneous determination of 22 mycotoxins in Pheretima aspergillum (E.Perrier) and Pheretima guillelmi (Michaelsen). A modified Quick Easy Cheap Effective Rugged and Safe (QuEChERS) method was used for sample preparation with recoveries ranging from 73% to 105% with relative standard deviations (RSDs) 0.99. In all cases, the intra- and inter-day precisions were lower than 6% and 10%, respectively. Matrix-matched calibration was utilized for quantification purposes to compensate for the matrix effects. Furthermore, the established method was successfully applied in 17 batches of normal real samples collected from different areas of China and 2 batches of moldy samples due to improper storage, only mold-contaminated samples were confirmed to have fumonisin B1 (FB1) and fumonisin B2 (FB2) contamination at 2.54-3.78μgkg-1. The constructed method could serve as a practical application of the UHPLC-MS/MS method for the trace analysis of multiple mycotoxins in complex matrixes, especially for those with high lipid contents.

In this study the occurrence of hidden fumonisin B1 (FB1) and fumonisin B2 (FB2) was analysed, on two cereal substrates (maize and rice), inoculated with Fusarium verticillioides (MRC 826), in order to determine the ratio of hidden FB1 and FB2. Two parallel methods were applied: an in vitro human digestion sample pre-treatment and the routine extraction procedure, in both cases with subsequent LC-MS analysis. It was found that all samples showed higher concentration of total fumonisin B1 after digestion, as compared to that of free fumonisin analysed only after extraction. The percentage of the hidden form by maize was 18.8 % (±2.4) for FB1 and 36.8 % (±3.8) for FB2, while for rice it was 32.3 % (±11.3) and 58.0 (±6.8), respectively, expressed as the proportion to total fumonisin B1, for the total dataset. Significant differences were found in the FB1 and FB2 concentration measured after the different digestion phases (saliva, gastric and duodenal) in case of both matrixes. The results are useful for human risk assessment, since both humans and animals may be exposed to markedly higher toxin load, as determined merely by conventional analytical methods.

A reliable and large-scale preparative method was developed for the simultaneous preparation of high-purity fumonisin B1 (FB1), fumonisin B2 (FB2) and fumonisin B3 (FB3) from maize culture inoculated with Fusarium moniliforme in this study. The targeted B-series fumonisins (FBs) were firstly extracted with acetonitrile/water (50 : 50, v/v) and then purified using MAX cartridges. The subsequent chromatographic preparative procedures on a Unitary C18 column (250 mm × 10 mm, 5 μm) and a SB-CN column (250 mm × 9.4 mm, 5 μm) were performed to effectively eliminate the other interferents to obtain the high-purity compounds. The prepared compounds were clearly identified to be FB1, FB2 and FB3 using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR). The purities were 96.25 ± 0.18%, 98.07 ± 0.18% and 97.36 ± 0.74% for FB1, FB2 and FB3, respectively, which were determined by the external standard LC-MS/MS method. Within 72 h, large amounts of high-purity FB1 (80 mg), FB2 (16 mg) and FB3 (16 mg) could be simultaneously prepared by the established approach.

The Aspergillus niger aggregate contains 15 morphologically indistinguishable species which presence is related to ochratoxin A (OTA) and fumonisin B2 (FB2) contamination of foodstuffs. The taxonomy of this group was recently reevaluated and there is a need of new studies regarding the risk that these species might pose to food security. 258 isolates of A. niger aggregate obtained from a variety of products from Spain were classified by molecular methods being A. tubingensis the most frequently occurring (67.5%) followed by A. welwitschiae (19.4%) and A. niger (11.7%). Their potential ability to produce mycotoxins was evaluated by PCR protocols which allow a rapid detection of OTA and FB2 biosynthetic genes in their genomes. OTA production is not widespread in A. niger aggregate since only 17% of A. niger and 6% of A. welwitschiae isolates presented the complete biosynthetic cluster whereas the lack of the cluster was confirmed in all A. tubingensis isolates. On the other hand, A. niger and A. welwitschiae seem to be important FB2 producers with 97% and 29% of the isolates, respectively, presenting the complete cluster. The genes involved in OTA and FB2 were overexpressed in producing isolates and their expression was related to mycotoxin synthesis.

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.

Cashew nut is a cash crop in tropical regions of Africa including Benin, providing substantial revenues to small-scale farmers and contributing significantly to the national gross domestic product. In Beninese context, about 95% of raw cashew nuts are exported. Therefore, the production of raw cashew nuts of good quality is mandatory to enter the international market. Moreover, health issues related to the consumption of nuts contaminated with mycotoxins are of high concern for human being. The objective of this research was to investigate the occurrence of mycotoxins and other secondary metabolites of Aspergillus sp. on raw cashew nuts and to evaluate farmers’ practices that affect the quality of raw nuts. In surface disinfected samples 90.2% of raw kernels were contaminated by strains of A. section Nigri in the northern guinea (NG) zone compared to 87.2% in the southern guinea (SS) zone. The level of contamination of raw cashew kernels by strains of A. section Flavi was 6.7% in the NG zone whereas it was 4.6% in the SS zone. When non disinfected kernels were plated, A. section Nigri was predominant in both NG and SS zones, with percentages of 89.7% and 93.4%, respectively. Ten and 15.7% of raw cashew kernels were contaminated by strains of A. section Flavi in NG and SS zones, respectively. A. flavus, A. tamari, A. costaricaensis, A. minisclerotigenes and A. nomius were identified as strains in A. section Flavi. The average water content and the cashew nut count were respectively 8.6% and 172 nuts/kg in the NG zone and 8.7% and 174 nuts/kg in the SS zone. The occurrence of aflatoxins, analysed by ultra-high performance liquid chromatography tandem mass spectrometry, showed that none of the analysed samples was positive for natural occurrence of aflatoxins with a detection limit of 0.05 - 0.2 µg/kg. All the above results showed that, at present, cashew nuts from Benin were in the range of good quality for export. Seven species of black aspergilli were isolated based on morphological and chemical identification namely A. tubingensis (44% of total strains), A. niger (with a chemical profile similar to A. welwitschiae) (32%), A. brasiliensis (10%), A. carbonarius (8.7%), A. luchuensis (synonym to A. kawachi or A. acidus) (2.7%), A. aculeatus (2%) and A. aculeatinus (0.7%). Forty five different metabolites and their isomers were identified and classified in 2 groups. Group A of known mycotoxins: fumonisin B2, fumonisin B4, ochartoxin B, ochratoxin A and secalonic acid A, secalonic acid D and secalonic acid F. Group B included other secondary metabolites like nigragillin, pyranonigrin A, nigerazine A, nigerazine B, nigerapyrone E, tensyuic acid A, tensyuic acid F, pyranonigrin B, pyranonigrin C, pyranonigrin D, fonsecin, tensidol A, pyrophen, atromentin, tensyuic acid B, funalenone, rubrofusarin, orlandin, asperazine, tensyuic acid C, tensyuic acid D, nigerasperone A, tensidol B (pestalamide A), fonsecin B, malformin A2, tubingensin A, tubingensin B, malformin C, kotanin, nominine, antafumicin A, antafumicin B, aurasperone C, aurasperone F, aurasperone E, aurasperone B, flavasperone and nafuredin. DNA analysis resulted in a phylogenetic trees based on combined sequences data of β-tubulin and calmodulin of 27 strains of A. section Nigri that clustered in 5 clades. Also, the chemical profile clustering showed also 5 groups confirming the species specific metabolites production. Sorting and grading resulted in two grades. First (good) grade (Grade 1) consisted of nuts with less than 50% of their shell surface affected by fungi and/or insect injuries, whereas second (bad) grade (Grade 2) were nuts that had 50% or more of their shell surface covered by fungi and/or insect symptoms. Nut counts in Grade 1 were from 151 to 174 nuts/kg and from 142 to 182 nuts/kg in NG and SS, respectively. In Grade 2, nut counts were from 168 to 202 nuts/kg in NG and from 171 to 197 nuts/kg in SS. These results indicated that sorting and grading resulted in Grade 1 of better quality with high nut count, and low fungal contamination. Interviews revealed the dominance of men (95%) in cashew nut cultivation indicating that it as a gendered activity. Cashew nut farming is labour intensive with 55% of farmers in the range of 35 to 55 years old. Unfortunately, most cashew farmers were less educated with up to 76% not exceeding primary educational level, making the educational level of farmers to be the main constrain for the production of cashew nut of good quality. This resulted of 80% of cashew farmers to select their seedlings from uncertified sources and 59% to never ask or received advice from extension services about cashew farming practices. Cashew nut farmers reported that cashew revenue contributed significantly to the household income for 44% and 54% of farmers in the NG and the SS zone, respectively. Other constrains for good cashew nut production were lack of sorting, drying and farm management. The farmer’s education level, limited access to extension services, lack of sorting, drying and farm management are major parameters that affect the quality of raw cashew nuts. Improvement of these parameters can positively affect raw nut quality and therefore contribute to better revenue from raw cashew production. Together with good farming and post-harvest management practices, the implementation of specific extension service for cashew growers and an incentive for sorting and grading as low cost technology, will impact the quality parameters of raw cashew nuts intended for export from Benin.

The present study was conducted to explore the occurrence of mycotoxins in commercial baby foods in Doha-Qatar. LCMS/MS- and HPLC-based analysis of baby food (n?=?67) for 12 mycotoxins confirmed the presence of aflatoxin M1 (AFM1, 33%), ochratoxin A (OTA, 31%), deoxynivalenol (DON, 27%), aflatoxin B1 (AFB1, 22%), fumonisin B2 (FB2, 10%), zearalenone (ZEN, 4%) and T-2 toxin (2%). Noodles exhibited the maximum contamination percentage, with 33% of the samples being contaminated above the EU maximum limits, for at least one mycotoxin. Among the multi-grain flake samples, up to 28% and for the milk and milk-based-cereal samples, 14% contained at least one mycotoxin above the EU maximum limits. From all cereal-based food samples, 22%, 5%, 2% and 2% were concurrently contaminated with 2, 3, 4 and 5 mycotoxins, respectively. The occurrence of toxicological important mycotoxins in Qatari market warrants the implementation of strict regulatory limits to protect human health. ? 2018, ? 2018 Informa UK Limited, trading as Taylor & Francis Group. This publication was made possible by NPRP grant # [NPRP8-392-4-003] from the Qatar National Research Fund (a member of the Qatar Foundation). Scopus

Fumonisin B1 (FB1) is a secondary metabolite produced by Fusarium verticillioides or Fusarium proliferatum, which present in food and feed. It causes hazardous effects on human and animal health. A monoclonal antibody (mAb) against FB1 was produced and a simple, reliable and sensitive, competitive, indirect enzyme-linked immunosorbent assay (ci-ELISA) for detection of FB1 was developed and the experiment conditions were optimized. The coating concentration of FB1-ovalbumin (FB1-OVA) was 500 ng mL−1, the action concentrations of anti-FB1 mAb and goat anti-mouse IgG were 1.28 × 104 and 1:5000, respectively. The 50% inhibitory concentration (IC50) was 11 ng mL−1, with a detectable range of 1.25–250 ng mL−1, and a limit of determination (LOD) of 1.15 ng mL−1. The cross-reactivity (CR) of the antibody against fumonisin B2 (FB2) was 60.4, and

Rationale Fumonisins and AAL-toxins are structurally similar mycotoxins that contaminate agricultural crops and foodstuffs. Traditional analytical screening methods are designed to target the known compounds for which standards are available but there is clear evidence that many other derivatives exist and could be toxic. A fast, semi-targeted method for the detection of all known fumonisins, AAL-toxins and related emerging toxins is required. Methods Strains of Fusarium verticillioides, Alternaria arborescens and Aspergillus welwitschiae were grown on their associated crops (maize, tomatoes, and grapes, respectively). Extracts were first analyzed in negative mode using product ion filtering to detect the tricarballylic ester product ion that is common to fumonisins and AAL-toxins (m/z 157.0142). During the same liquid chromatography (LC) run, rapid polarity switching was then used to collect positive mode tandem mass spectrometric (MS2) data for characterization of the detected compounds. Results Fumonisin B1, B2, B3 and B4 were detected on Fusarium contaminated maize, AAL-toxins TA, TB, TD, TE were detected on Alternaria inoculated tomatoes and fumonisin B2, B4 and B6 on Aspergillus contaminated grapes. Additionally, over 100 structurally related compounds possessing a tricarballylic ester were detected from the mould inoculated plant material. These included a hydroxyl-FB1 from F. verticillioides inoculated maize, keto derivatives of AAL-toxins from A. arborescens inoculated tomatoes, and two previously unreported classes of non-aminated fumonisins from Asp. welwitschiae contaminated grapes. Conclusions A semi-targeted method for the detection of all fumonisins and AAL-toxins in foodstuffs was developed. The use of the distinctive tricarballylic ester product anion for detection combined with rapid polarity switching and positive mode MS2 is an effective strategy for differentiating between known isomers such as FB1 and FB6. This analytical tool is also effective for the identification of new compounds as evident from the discoveries of the previously unreported hydroxyl-FB1, keto-AAL-toxins, and the two new families of non-aminated fumonisins. © 2015 Her Majesty the Queen in Right of Canada Rapid Communications in Mass Spectrometry © 2015 John Wiley & Sons Ltd. Reproduced with the permission of the Ministers of Agriculture and Agri-Food Canada.

Pre- and postharvest fruit rots of fungal origin are an important burden for the pomegranate industry worldwide, affecting the produce both quantitatively and qualitatively. During 2013, local orchards were surveyed and 280 fungal isolates from Greece (GR) and Cyprus (CY) were collected from pomegranates exhibiting preharvest rot symptoms, and additional 153 isolates were collected postharvest from cold-stored fruit in GR. Molecular identification revealed that preharvest pomegranate fruit rots were caused predominately by species of the genera Aspergillus (Aspergillus niger and Aspergillus tubingensis) and Alternaria (Alternaria alternata, Alternaria tenuissima, and Alternaria arborescens). By contrast, postharvest fruit rots were caused mainly by Botrytis spp. and to a lesser extent by isolates of Pilidiella granati and Alternaria spp. Considering that a significant quota of the fungal species found in association with pomegranate fruit rots are known for their mycotoxigenic capacity in other crop systems, their mycotoxin potential was examined. Alternariol (AOH), alternariol monomethyl-ether (AME) and tentoxin (TEN) production was estimated among Alternaria isolates, whereas ochratoxin A (OTA) and fumonisin B2 (FB2) production was assessed within the black aspergilli identified. Overall in both countries, 89% of the Alternaria isolates produced AOH and AME in vitro, while TEN was produced only by 43.9%. In vivo production of AOH and AME was restricted to 54.2% and 31.6% of the GR and CY isolates, respectively, while none of the isolates produced TEN in vivo. Among black aspergilli 21.7% of the GR and 17.8% of the CY isolates produced OTA in vitro, while in vivo OTA was detected in 8.8% of the isolates from both countries. FB2 was present in vitro in 42.0% of the GR and 22.2% of the CY isolates, while in vivo the production was limited to 27.5% and 4.5% of the GR and the CY isolates, respectively. Our data imply that mycotoxigenic Alternaria and Aspergillus species not only constitute a significant subset of the fungal population associated with pomegranate fruit rots responsible for fruit deterioration, but also pose a potential health risk factor for consumers of pomegranate-based products.

Mycotoxin producing moulds may contaminate numerous agricultural commodities either before harvest or during storage. A varied diet consisting of different foods may therefore be contaminated with a range of mycotoxins. The aim of the present study was to study concurrent exposure to mycotoxins through urinary multi-biomarker analysis, as well as its possible associations with the diet.Urinary samples from 252 adults, participating in the Swedish national dietary survey Riksmaten 2010-11, were collected together with a 4-day diet record. Concurrent mycotoxin exposure was studied using a multi-biomarker LC-MS/MS method. The results revealed that exposure to mycotoxins is common and concurrent exposure to more than one toxin was found in 69% of the study population. However, when comparing the number of toxins detected with the reported consumption data it was difficult to distinguish food patterns which would indicate an increased risk of exposure to many mycotoxins simultaneously.This is the first study to investigate concurrent mycotoxin exposure and urinary levels of fumonisin B1 (FB1), fumonisin B2 (FB2), nivalenol (NIV), ochratoxin A (OTA), zearalenone (ZEA), ?-zearalenol (?-ZOL), ?-zearalenol (?-ZOL) and de-epoxydeoxynivalenol (DOM-1) among adults in Sweden.

BACKGROUND Aspergillus niger has been reported as a potentially dangerous pathogen of date-palm fruits in Saudi Arabia due to the production of fumonisin B2 (FB2) and ochratoxin A (OTA). In a trial to disinfect this product, a double atmospheric pressure argon cold plasma (DAPACP) jet system was set up and evaluated against spore germination and mycotoxin production of the pathogen. RESULTS The plasma jets were characterised photographically, electrically and spectroscopically. DAPACP jet length increases with the increase of argon flow rate, with optimum rate at 3.5 L min−1. The viability of A. niger spores, inoculated onto sterilised date palm fruit discs, progressively decreases with extension of the exposure time of DAPACP due to the more quantitative amount of OH and O radicals interacting with the examined samples. There was a progressive reduction of the amount of FB2 and OTA detected in date palm discs on extension of the exposure time of the plasma-treated inoculums at flow rate of 3.5 L min−1. FB2 was not detected in the discs inoculated with 6-min plasma-treated A. niger, while OTA was completely absent when the fungus was treated for 7.5 min. CONCLUSION DAPACP showed promising results in dry fruit decontamination and in inhibition of mycotoxin release by A. niger contaminating the fruits. The progress in the commercial application of cold plasma needs further investigation concerning the ideal width of the plasma output to enable it to cover wider surfaces of the sample and consequently inducing greater plasma performance © 2014 Society of Chemical Industry

In this study a multi-mycotoxin method was applied to screen seventeen chemically divergent mycotoxins from dry chilli samples (Capsicum annum L.) collected from the markets in Sri Lanka (n ¼ 86) and Belgium (n ¼ 35) using a high performance liquid chromatography tandem mass spectrometer. In addition to aflatoxins (

A quantitative risk assessment of mycotoxins due to the consumption of chilli (Capsicum annum L.) and black pepper (Piper nigrum L.) was performed in Sri Lanka. A food frequency questionnaire was administered in order to collect the data on consumption of spices by households in the Northern and Southern region (n = 249). The mean chilli consumption in the North was significantly higher (p < 0.001) compared to the South. Mean exposure to aflatoxin B1 (AFB1) in the North (3.49 ng/kg BW/day) and South (2.13 ng/kg BW/day) have exceeded the tolerable daily intake due to chilli consumption at the lower bound scenario, while exposure to OTA was small. Dietary exposure to other mycotoxins, fumonisin B1, fumonisin B2, sterigmatocystin and citrinin due to spices were estimated. Margin of exposure estimations at the mean exposure to AFB1 were remarkably lower due to chilli (45–78) than for pepper (2315–10,857). Moreover, the hepato cellular carcinoma (HCC) risk associated with the mean AFB1 exposure through chilli at the lower bound was 0.046 and 0.028 HCC cases/year/100,000 based on the North and South consumption, respectively. AFB1 exposure via chilli should be considered as a great public health concern in Sri Lanka due to both high mycotoxin concentration and high consumption.

Most tropical fruits after harvest are very perishable because of fungal infection. Since some pathogenic fungi can produce hazardous compounds such as mycotoxins, novel rapid and effective methods to assess those hazardous compounds are urgently needed. Herein we report that Vibrio qinghaiensis sp. Q67, a luminescent bacterium, can be used to rapidly assess the toxicities of mycotoxins and cultures from mycotoxin-producing pathogens. A good correlation (R2 > 0.98) between concentrations of the mycotoxins (fumonisin B1, deoxynivalenol, zearalenone, ochratoxin A, patulin, and citrinin) and the luminous intensity of V. qinghaiensis sp. Q67 was obtained. Furthermore, significant correlations (R2 > 0.96) between the amount of mycotoxin and the luminous intensity from the cultures of 10 major mycotoxin-producing pathogens were also observed. In addition, Fusarium proliferatum (half-maximal inhibitory concentration (IC50) = 17.49%) exhibited greater luminescence suppression than Fusarium semitectum (IC50 = 92.56%) or Fusarium oxysporum (IC50 = 28.61%), which was in agreement with the existing higher levels of fumonisin B1, fumonisin B2, and deoxynivalenol, which were measured by high-performance liquid chromatography-tandem mass spectrometry. These results suggest that V. qinghaiensis sp. Q67 is a promising alternative for the rapid evaluation of the toxicity of fungal mycotoxins.

Several plant pathogen Fusarium species produce fumonisins (FUMs); which can end up in food and feed and; when ingested; can exhibit harmful effects on humans and livestock. Mycotoxin intoxication by fumonisin B1 (FB1) and fumonisin B2 (FB2) can cause porcine pulmonary edema; leukoencephalomalacia in equines; esophageal cancer and birth defects by natural contamination. Herein; the occurrence of FB1 and FB2 in feedstuff (compound feed and feed ingredients) was investigated between 2011 and 2016 in South Korea. A total of 535 animal feed samples (425 compound feed samples and 110 feed ingredients) produced domestically were sampled four times between 2011 and 2016 (2011; 2012; 2014 and 2016) from feed factories in South Korea. The limit of detection (LOD) for FB1 and FB2 was 20 μg/kg and 25 μg/kg; respectively; and the limit of quantitation (LOQ) was 30 μg/kg and 35 μg/kg; respectively. The recovery range (%) was between 86.4% and 108.8%; and the relative standard deviation (RSD) (%) was 4.7–12.1%. Seven (swine feed samples) out of the 425 feed samples exceeded the European Union (EU) and South Korea commission regulations over the six-year test period; and no feed ingredients exceeded the guidelines.

There are a total of 30 strains including 27 strains of Aspergillus niger intended used in Chinese food industry, two strains used as control and one strain isolated from corn for fumonisin (FB) production on 3 media. It was found that FB2 production by A. niger was function-dependent and highly related to culture media, as well as incubation time. All strains studied were unable to produce FB1 and FB3. Almost all strains were found to produce FB2 on corn, rice and wheat bran. Based on their intended use in the food industry, the higher level of FB2 producers were strains used for saccharifying enzyme (n = 13) production, followed by organic acid (n = 6), tannase (n = 7) and β-galactosidase (n = 1) production, with the FB2 mean level of 3553–10,270 μg/kg, 1059–12,036 μg/kg, 3–7 μg/kg and 2–4 μg/kg on corn, 5455–9241 μg/kg, 559–2190 μg/kg, 4–9 μg/kg and 6–10 μg/kg on rice, 5959–7709 μg/kg, 9491–17,339 μg/kg, 8–14 μg/kg and 120–222 μg/kg on wheat bran, respectively. Comparatively, strains of Fusarium verticillioide were capable of producing fumonins simultaneously with broader spectrum including FB1, FB2 and FB3, but at a much lower level. In conclusion, it is necessary to evaluate FB2 production by A. niger before intended use in the food processing industry.

Bertholletia excelsa is the tree that produces Brazil nuts which have vast economic importance in the Amazon region and as an export commodity. The aim of this study was to assess the presence of Aspergillus section Nigri in Brazil nut samples at different stages of its production chain and to verify the toxigenic potential for fumonisin B2 (FB2) production of these isolates along with the presence of this mycotoxin in Brazil nut samples. The fungal infection ranged from 0 to 80% at the different stages of the harvest and processing chain and the water activity of the nuts from 0.273 to 0.994. A total of 1052 A. section Nigri strains were isolated from Brazil nuts and 200 strains were tested for their ability to produce FB2: 41 strains (20.5%) were FB2 producers with concentrations ranging from 0.09 to 37.25 mg/kg; 2 strains (1%) showed traces of FB2, less than the detection limit (0.08 mg/kg); and 157 (78.5%) were not FB2 producers. Although several samples showed high contamination by A. section Nigri, no sample was contaminated by FB2.