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Enniatin B (ENNB) is a mycotoxin produced by moulds from the Fusarium genera and its toxic effects are still not fully elucidated, hence a safe reference exposure value has not been established yet. ENNB is the most prevalent emerging mycotoxin and is widely found in cereal-based products, nevertheless, there are no comprehensive exposure assessment studies. For that reason, the aim of this study was to characterise the occurrence of ENNB and estimate the exposure of the Spanish and Catalan populations. A total of 347 cereal-based products were collected in 2019 and were analysed using liquid chromatography-tandem mass spectrometry. Consumption data were obtained from the national food consumption surveys (ENALIA) and a regional survey conducted in Catalonia. The global exposure was estimated using deterministic and probabilistic methods. The results showed a high occurrence of close to 100% in all foodstuffs, with a range from 6 to 269 μg/kg, and a strong correlation with the levels of deoxynivalenol. Children aged one-nine years were the most exposed, showing mean estimates in the range 308-324 ng/kg bw/day and 95th percentiles 697-781 ng/kg bw/day. This study stresses the need for further toxicological data to establish reference doses and conclude formal risk assessment, accounting for the co-occurrence with deoxynivalenol. This research was funded by the Catalan Agency of Food Safety (ACSA) (SA-2019-391) and the Agency for Management of University and Research Grants (AGAUR) with a doctoral fellowship (2021 FI_B 00129; 2022 FI_B1 00033).

In a temperate climate, the mycotoxins deoxynivalenol (DON), nivalenol (NIV), enniatin B (ENN B), mycophenolic acid (MPA), roquefortine C (ROC) and zearalenone (ZEN) are often found in maize silage. Although rumen microbiota are able to degrade some mycotoxins (e.g. DON), others are known to stay mainly intact (e.g. ROC). In addition, mycotoxin degradation can be hampered by a low ruminal pH or decrease in rumen microbial activity. Hence, these mycotoxins can reach the small intestine and exert a cytotoxic effect on intestinal epithelial cells. In this study, a real-time kinetic analysis of the cytotoxicity of these six mycotoxins and some of their metabolites (α- and β-zearalenol, α- and β-ZOL) was performed in a calf small intestinal epithelial cell line (CIEB). Confluency as well as the cell death parameters apoptosis and necrosis were determined to evaluate the mycotoxin-induced cytotoxicity. A combination of Annexin-V green and Cytotox red staining was used to determine early and late apoptosis as well as necrosis. Six different concentrations were tested ranging from 0.78 to 12.5 μM. Compared to cells not exposed to mycotoxins, DON and NIV exert a fast toxic effect with DON being more toxic than NIV within the first hours of incubation, whereas the inverse was observed at 16 h of incubation. On the other hand, MPA and ZEN induced increased Annexin V green positive cells within several hours of incubation with higher toxicity over time. Increased Annexin V green and Cytotox red positive cells were seen for ROC only at the highest concentration tested. For ENN B, increased Annexin V green positive cells were observed only after 12 h and α- and β-ZOL did not show cytotoxic effects. Hence, mycotoxin exposure causes either severe (DON and NIV) or more limited (ZEN, ROC, MPA, and ENN B) risk of bovine intestinal epithelial damage.

In the period 2009–2018, an annual field survey with commercial arable farms in the Netherlands was held, to collect data on agronomics of wheat fields as well as mycotoxin concentrations of the specific wheat field at harvest. In total, 293 full farm field records over 8 years were obtained. This study aimed to investigate (i) the occurrence of deoxynivalenol (DON) and other mycotoxins, as well as correlations between these mycotoxins, and (ii) the relationships between agronomics and the DON concentration in wheat kernels for wheat cultivated in the Netherlands. Results showed that mycotoxins most frequently observed in concentrations above the limit of quantification were DON, enniatin B and B1, HT-2 toxin, zearalenone (ZEN) and nivalenol. On average, DON was detected in 54% of the samples (> 50 µg/kg) ranging from 19 to 92% depending on the year. Positive samples (> 50 µg/kg) had DON concentrations ranging 53–15,400 µg/kg, with a median of 228 µg/kg. Co-occurrence between DON and ZEN as well as between each of DON and ZEN with their modified forms was confirmed by the data of this study. The year influenced the DON concentration in wheat the most, followed by the region. The results of this study show that DON levels in wheat can only be influenced in a limited manner by agronomic practices such as the use of fungicides against Fusarium spp. around flowering, crop rotation, or the use of resistant wheat cultivars.

A number of 344 samples were collected from Tianjin, China and were analysed for the occurrence of emerging Fusarium mycotoxins including enniatin A (ENN A), enniatin A1 (ENN A1), enniatin B (ENN B...

Twenty-two mycotoxins were investigated in bread and biscuits samples (n = 115) purchased from Tunisia. Deoxynivalenol (DON), enniatin B (ENB), ENB1, and zearalenone (ZEA) occurred in the samples with ENB detected in all the samples. Whole bread was the most contaminated matrix showing the highest amounts of DON (25.2 ng/g), ENB (10.7 ng/g), and ENB1 (5.4 ng/g). However, biscuits showed a maximum level of ZEA (1.08 ng/g). Co-occurrence were also evaluated, 65% of biscuit samples were contaminated by three mycotoxins simultaneously (DON + ENB + ENB1) and the most frequent combination (DON + ENB) co-occurred in 83% of the white bread samples and 78% of the whole bread samples. DON dietary exposure was estimated to be 15.2 and 11 ng/kg bw/day for bread and biscuits, respectively for adults. While for children, 38.7 and 81 ng/kg bw/day, respectively. However, no real health risk was found, and children are having up to 12% of the tolerable daily intake (TDI) of DON and adults up to 2.6% of the DON TDI through the consumption of both cereal products.

The extensive use of plant ingredients in novel aquafeeds have introduced mycotoxins to the farming of seafood. The emerging enniatin B (ENNB) and beauvericin (BEA) mycotoxins have been found in the novel aquafeeds and farmed fish. Little is known about the potential toxicity of ENNs and BEA in farmed fish and their feed-to-organ transfer. Atlantic salmon (Salmo salar) presmolt (75.3 +/- 8.10 g) were fed four graded levels of spiked chemical pure ENNB or BEA feeds for three months, in triplicate tanks. Organismal adverse health end-point assessment included intestinal function (protein digestibility), disturbed hematology (red blood cell formation), bone formation (spinal deformity), overall energy use (feed utilization), and lipid oxidative status (vitamin E). Both dietary BEA and ENNB had a low ( liver > brain > muscle), with a higher transfer for ENNB compared to BEA. BEA caused a growth reduction combined with a decreased protein digestion and feed conversion rate-ENNB caused a stunted growth, unrelated to feed utilization capacity. In addition, ENNB caused anemia while BEA gave an oxidative stress response. Lower bench-mark dose regression assessment showed that high background levels of ENNB in commercial salmon feed could pose a risk for animal health, but not in the case of BEA. Grant 281032 HAVBRUK2; info:eu-repo/semantics/publishedVersion

Food contaminants of bacterial or fungal origin frequently contaminate staple foods to various extents. Among others, the bacterial toxin cereulide (CER) and the mycotoxin deoxynivalenol (DON) co-occur in a mixed diet and are absorbed by the human body. Both toxins exert dis-tinctive mitotoxic potential. As damaged mitochondria are removed via autophagy, mitochondrial and lysosomal toxicity were assessed by applying low doses of single and combined toxins (CER 0.1–50 ng/mL; DON 0.01–5 µg/mL) to HepG2 liver cells. In addition to cytotoxicity assays, RT-qPCR was performed to investigate genes involved in lysosomal biogenesis and autophagy. CER and DON caused significant cytotoxicity on HepG2 cells after 5 and 24 h over a broad concentration range. CER, alone and in combination with DON, increased the transcription of the autophagy related genes coding for the microtubule associated protein 1A/1B light chain 3 (LC3) and sequestome 1 (SQSTM1) as well as LC3 protein expression which was determined using immunocytochemistry. DON increased LC3 protein expression without induction of gene transcription, hence it seems plausible that CER and DON act on different pathways. The results support the hypothesis that CER induces autophagy via the LC3 pathway and damaged mitochondria are therefore eliminated.

Food and feed contamination by nonlegislated mycotoxins beauvericin (BEA) and enniatin B (ENB) is a worldwide health concern in the present. The principal objective of this work is to assess some of the existing protocols to discover the single nucleotide variants (SNVs) in transcriptomic data obtained by RNA-seq from Jurkat cells in vitro samples individually exposed to BEA and ENB at three concentration levels (1.5, 3 and 5 µM). Moreover, previous transcriptomic results will be compared with new findings obtained using a different protocol. SNVs rs201003509 in BEA exposed cells and the rs36045790 in ENB were found in the differentially expressed genes in all doses compared to controls by means of the Genome Analysis Toolkit (GATK) Best Practices workflow. SNV-RNA-seq complementary pipeline did not show any SNV. Concerning gene expression, discrepant results were found for 1.5 µM BEA exposed cells compared with previous findings. However, 354 overlapped differentially expressed genes (DEGs) were identified in the three ENB concentrations used, with 147 matches with respect to the 245 DEGs found in the previous results. In conclusion, the two discovery SNVs protocols based on variant calling from RNA-seq used in this work displayed very different results and there were SNVs found manually not identified by any pipeline. Additionally, the new gene expression analysis reported comparable but non identical DEGs to the previous transcriptomic results obtained from these RNA-seq data. Single-nucleotide variants discovery protocols in transcriptomic data by RNA-seq from Jurkat cells exposed to mycotoxins for the first time Single-nucleotide variants discovery protocols in transcriptomic data by RNA-seq from Jurkat cells exposed to mycotoxins for the first time

Wheat is one of the world’s key staple foods, but it is often contaminated with mycotoxin-producing microorganisms, resulting in a large amount of food waste every year. The contamination of wheat grains harvested in 2020 and 2021 in Switzerland, as well as of wheat flours bought in local stores with the two mycotoxins zearalenone (ZEA) and enniatin B (ENB) was investigated. The quantification was performed using LC–MS/MS. ZEA, the level in different cereals and food products of which is regulated by law, was detected in half of the grain samples at levels below 100 µg/kg, except for one sample contaminated with 147 µg/kg. No ZEA was detected in the commercial wheat flours. The emerging mycotoxin ENB was detected in all samples of wheat grains and flours, at levels between 3 and 938 µg/kg. The harvest year was shown to affect the ENB content (p value < 0.01), and in particular the humid weather conditions encountered in 2021 during the month of harvest. The refining grade of the flours showed no influence on the contamination by ENB, indicating that the contamination with ENB can occur not only on the surface layers but also on the inner layers on the wheat grain. As chronic exposure to ENB can therefore not be excluded, decontamination solutions are needed to prevent food waste and further improve the food safety of wheat-based products.

Enniatin B (ENN B) and Beauvericin (BEA) are cyclohexadepsipeptides that can be isolated from Fusarium and Beauveria bassiana, respectively. Both compounds are cytotoxic and ionophoric. In the present study, the mechanism of cell death induced by these compounds was investigated. Epidermal carcinoma-derived cell line KB-3-1 cells were treated with different concentrations of these compounds. The extracellular secretion of cathepsin B increased in a concentration-dependent manner, and the lysosomal staining by lysotracker red was reduced upon the treatment with any of the compounds. However, the extracellular secretion of cathepsin L and cathepsin D were not affected. Inhibition of cathepsin B with specific inhibitor CA074 significantly reduced the cytotoxic effect of both compounds, while inhibition of cathepsin D or cathepsin L did not influence the cytotoxic activities of both compounds. In vitro labelling of lysosomal cysteine cathepsins with Ethyl (2S, 3S)-epoxysuccinate-Leu-Tyr-Acp-Lys (Biotin)-NH2 (DCG04) was not affected in case of cathepsin L upon the treatment with both compounds, while it was significantly reduced in case of cathepsin B. In conclusion, ENN B and BEA increase lysosomal Ph, which inhibits delivery of cathepsin B from Golgi to lysosomes, thereby inducing cathepsin B release in cytosol, which activates caspases and hence the apoptotic pathway.

The exposure to mycotoxins of Swedish adolescents is currently unknown. The aim of the present study was to investigate the exposure to mycotoxins and their association with food intake, and background characteristics in adolescents of a national dietary survey. About 3000 school students (1000 from the 5th, 8th and 11th school years) were recruited for the survey. The participants completed Web-based questionnaires on food propensity, sociodemography and health, and a Web-based dietary recall. Spot urine and blood samples were collected from 1105 of the participants for mycotoxin biomarker analysis. Mycotoxins were analysed with multibiomarker methods in urine (HPLC-MS/MS) and serum (HPLC-MS/MS). Of the 35 different analytes in urine, the frequency of positive samples were the following: deoxynivalenol (DON, 4.8%), DON-15-β-D-O-glucuronide (DON-15GlcA, 9.1%), dihydro-citrinone (DH-CIT, 0.5%), HT-2-glucuronide (HT-2-3-GlcA, 0.1%) and ochratoxin A (OTA, 0.1%). Of the 27 different analytes in serum, OTA was detected in all samples, while 2’R-ochratoxin A (2’R-OTA) was found in 8.3% and enniatin B (EnB) in 99.2% of the samples. Exposure assessment calculations were performed on OTA from the serum concentration and on DON equivalents (DON eqv) from the urine concentration. All probable daily intake (PDI) estimates were below tolerable daily intakes, except for 1.6% of the participants for DON. The maximum PDI was 4.3 μg DON eqv/kg body weight and day. Consumption of cereal grain commodities was associated with levels of DON, EnB or OTA in biofluids. Serum OTA was also associated with intakes of raisins and coffee. Furthermore, coffee consumption correlated well with 2’R-OTA concentration in serum. In conclusion, exposure to mycotoxins in Swedish adolescents is common, but fortunately, high exposure was rare. Electronic supplementary material The online version of this article (10.1007/s12550-019-00381-9) contains supplementary material, which is available to authorized users.

Contamination with enniatins A, A1, B and B1 was investigated in 1240 samples of small grain cereals (470 wheat, 260 durum wheat, 282 spring barley, 172 triticale and 56 winter barley) from the French harvests of 2012 to 2014. Associations with Fusarium avenaceum, F. tricinctum and F. poae were assessed, with the identification of Fusarium species by real-time PCR and mycotoxin quantification by LC–MS/MS. Enniatins were common in the fields sampled. Enniatin concentrations varied between years but were consistently highest on spring barley (mean values of 199 to 1316 μg/kg) and triticale (mean values from 131 to 1218 μg/kg), and lower on wheat (mean values from 47 to 142 μg/kg) and durum wheat (mean values from 55 to 596 μg/kg). The concentrations of the various enniatins were strongly correlated with each other (Pearson’s correlation coefficient of 0.61 to 0.98). Enniatin B was the most frequent (68% of the total enniatin content), followed by enniatin B1 (22%), enniatin A1 (7%) and enniatin A (3%). Fusarium species were quantified by calculating arithmetic mean total DNA levels. F. tricinctum was the most abundant (0.177 pg/ng total DNA), followed by F. avenaceum (0.141 pg/ng total DNA) and F. poae (0.091 pg/ng total DNA). Total DNA levels for each species, and the predominant species varied between years and crops. Small grain cereal species (p value

The occurrence of 12 mycotoxins has been analysed by liquid chromatography - time of flight mass spectrometry in the batch of 60 herbal teas purchased from drugstores in Latvia. Among the dry tea samples, 90% were positive for one to eight mycotoxins. Enniatin B and deoxynivalenol (DON) were the most frequently detected mycotoxins in 55% and 45% of the samples, respectively. DON reached the highest level, from 129 µg kg-1 in herbal blend to 5,463 µg kg-1 in wormwood tea. Ochratoxin A (OTA) and aflatoxin B1 (AFB1) were found in 10% and 20% of the samples at the concentrations ranged between 2.99-30.3 µg kg-1 and 3.40-23.7 µg kg-1. Studies of the tea infusion process indicated that 32-100% of DON and zearalenone present in dry teas were extracted into the infusions. Dietary exposure assessment was performed, using the determined mycotoxin levels and the available consumption data.

Increasing consumption of cereals has been associated with reduced risk of several chronic diseases, as they contain phytochemicals that combat oxidative stress. Cereal contamination by the “emerging mycotoxins” beauvericin (BEA) and enniatins (ENs) is a worldwide health problem that has not yet received adequate scientific attention. Their presence in feeds represents a risk for animals and a potential risk for humans because of their carry-over to animal-derived products. This preliminary study aimed to investigate if the total antioxidant capacity (TAC) of corn, barley, and wheat flours could be influenced by contamination with increasing levels of BEA and ENN B. The highest TAC value was observed in barley compared with wheat and corn (p < 0.001) before and after contamination. No effect of mycotoxin or mycotoxin level was found, whereas cereal x mycotoxin exhibited a significant effect (p < 0.001), showing a lower TAC value in wheat contaminated by ENN B and in barley contaminated by BEA. In conclusion, barley is confirmed as a source of natural antioxidants with antiradical potentials. Additional studies with a larger sample size are necessary to confirm the obtained results, and investigations of the toxic effects of these emergent mycotoxins on animals and humans should be deepened.

The development of innovative food processing technologies has increased to answer the growing demand to supply of fresh-like products. The aim of the present study is to investigate the effect of pulsed electric fields (PEF) technology on reducing the emerging mycotoxins (enniatins (ENs) and beauvericin (BEA)) contents in juice and smoothie samples. The products of degradation obtained after PEF treatment were identified and their toxicological endpoint toxicities predicted by Pro Tox-II web. Mycotoxin reduction ranged from 43 to 70% in juices and smoothies, but in water the expected effect was lower. The acidified pH increased BEA reduction in water. The degradation products that were produced were the result of the loss of aminoacidic fragments of the original molecules, such as HyLv, Val, Ile, or Phe. Pro Tox-II server assigned a toxicity class I for enniatin B (ENB) degradation products with a predicted LD50 of 3 mg/Kgbw. The other degradation products were classified in toxicity class III and IV.

Cereal grain contaminated by Fusarium mycotoxins is undesirable in food and feed because of the harmful health effects of the mycotoxins in humans and animals. Reduction of mycotoxin content in grain by cleaning and size sorting has mainly been studied in wheat. We investigated whether the removal of small kernels by size sorting could be a method to reduce the content of mycotoxins in oat grain. Samples from 24 Norwegian mycotoxin-contaminated grain lots (14 from 2015 and 10 from 2018) were sorted by a laboratory sieve (sieve size 2.2 mm) into large and small kernel fractions and, in addition to unsorted grain samples, analyzed with LC-MS-MS for quantification of 10 mycotoxins. By removing the small kernel fraction (on average 15% and 21% of the weight of the samples from the two years, respectively), the mean concentrations of HT-2+T-2 toxins were reduced by 56% (from 745 to 328 &micro, g/kg) in the 2015 samples and by 32% (from 178 to 121 &micro, g/kg) in the 2018 samples. Deoxynivalenol (DON) was reduced by 24% (from 191 to 145 &micro, g/kg) in the 2018 samples, and enniatin B (EnnB) by 44% (from 1059 to 594 &micro, g/kg) in the 2015 samples. Despite low levels, our analyses showed a trend towards reduced content of DON, ADON, NIV, EnnA, EnnA1, EnnB1 and BEA after removing the small kernel fraction in samples from 2015. For several of the mycotoxins, the concentrations were considerably higher in the small kernel fraction compared to unsorted grain. Our results demonstrate that the level of mycotoxins in unprocessed oat grain can be reduced by removing small kernels. We assume that our study is the first report on the effect of size sorting on the content of enniatins (Enns), NIV and BEA in oat grains.

Introduction A mini-study of 20 raw milk samples was conducted to examine the spectrum of fungal metabolites in sheep milk from the first spring milking. Material and Methods Samples were collected from randomly selected ewes in two animal flocks from the Bieszczady Mountains and analysed using liquid chromatography-tandem mass spectrometry. Results Out of ~700 bacterial, fungal, and plant metabolites tested for, only one mycotoxin – Enniatin B – was detected in sheep milk samples (18/20; 0.0055–0.0121 μg/kg; 0.0078 μg/kg average). Conclusions The results indicated that there was no high-level exposure to fungal metabolites via consumption of raw sheep milk during the sample collection period.

In this study, the presence of fourteen Fusarium mycotoxins, legislated by the European Union – deoxynivalenol, zearalenone, HT-2 and T-2 toxins (EC/1881/2006; 2013/165/EU), or non-legislated (five trichothecens and five “emerging” mycotoxins), was evaluated in 31 whole unprocessed wheat samples and 35 white wheat flour samples from different areas of Romania. For this purpose, a validated multi-mycotoxins liquid chromatography tandem mass spectrometry method was applied. Seventy three percent of the analyzed samples contained at least one mycotoxin. The highest occurrence was for enniatin B, 71% of the analyzed samples being positive (21–407 μg kg−1). Regarding the legislated mycotoxins, deoxynivalenol was detected in 14% (111–1787 μg kg−1) of the samples, while zearalenone was detected in 9% (51–1135 μg kg−1). Only one sample was positive for neosolaniol. Concerning the co-occurrence, 42% of the samples were contaminated with two to five mycotoxins, the most frequent being the binary or tertiary combinations of enniatins. This is the first study applied to Romanian wheat grains and flour samples using a high sensitive multi-mycotoxins method, and which included also “emerging” mycotoxins.

Concern about the risk of exposure to emerging plant-derived mycotoxins such as beauvericin and enniatins has been addressed by the European Commission who requested the European Food Safety Authority for a scientific opinion on their risk to human and animal health. The studied mycotoxins were found in feeds with enniatin B and beauvericin at average concentrations of 19.9 μg/kg and 30 μg/kg, respectively. In all cases, concentrations of all the mycotoxins analyzed were below quantification limits (, AQUAEXCEL 2020 (652831) under TNA project AE050057: “Implications for aquaculture of non-regulated emerging mycotoxins” granted to J. Nácher-Mestre

Extraction experiments in the two-phase water-nitrobenzene system and γ-activity measurements were used to determine the stability constant of the enniatin B–K+ complex dissolved in nitrobenzene saturated with water as 105.5±0.2. Quantum mechanical calculations were applied to predict the most probable structure of this cationic complex species.

Forages are important components of dairy cattle rations but might harbor a plethora of mycotoxins. Ruminants are considered to be less susceptible to the adverse health effects of mycotoxins, mainly because the ruminal microflora degrades certain mycotoxins. Yet, impairment of the ruminal degradation capacity or high ruminal stability of toxins can entail that the intestinal epithelium is exposed to significant mycotoxin amounts. The aims of our study were to assess i) the mycotoxin occurrence in maize silage and ii) the cytotoxicity of relevant mycotoxins on bovine intestinal cells. In total, 158 maize silage samples were collected from European dairy cattle farms. LC-MS/MS-based analysis of 61 mycotoxins revealed the presence of emerging mycotoxins (e.g. emodin, culmorin, enniatin B1, enniatin B, and beauvericin) in more than 70% of samples. Among the regulated mycotoxins, deoxynivalenol and zearalenone were most frequently detected (67.7%). Overall, 87% of maize silages contained more than five mycotoxins. Using an in vitro model with calf small intestinal epithelial cells B, the cytotoxicity of deoxynivalenol, nivalenol, fumonisin B1 and enniatin B was evaluated (0&ndash, 200 &micro, M). Absolute IC50 values varied in dependence of employed assay and were 1.2&ndash, 3.6 &micro, M, 0.8&ndash, 1.0 &micro, M, 8.6&ndash, 18.3 &micro, M, and 4.0&ndash, 6.7 &micro, M for deoxynivalenol, nivalenol, fumonisin B1, and enniatin B, respectively. Results highlight the potential relevance of mycotoxins for bovine gut health, a previously neglected target in ruminants.

Totals of 158 corn and corn-based samples and 291 wheat and wheat-based samples from Shandong province, China in 2017 were analyzed for five mycotoxins including beauvericin (BEA), enniatin A (ENA), enniatin A1 (ENA1), enniatin B (ENB), and enniatin B1 (ENB1) by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). BEA was the predominant toxin detected, followed by ENB, ENA1, ENA, and ENB1. Corn and corn-based samples were more easily contaminated by BEA with an average concentration of 65.26 &micro, g/kg, compared with that in wheat and wheat-based samples (average = 0.41 &micro, g/kg). Concentrations of BEA, ENA, and ENB1 in corn kernels, flours, and flakes were significantly different (Kruskal&ndash, Wallis Test, p &lt, 0.05), as well as for BEA, ENA, ENB, and ENB1 in wheat kernels, flours, and noodles (Kruskal&ndash, Wallis test, p &lt, 0.05). Furthermore, 59.5% (94/158) and 59.8% (174/291) corn- and wheat-based samples were co-contaminated by at least two mycotoxins, respectively. Positive correlations in concentrations were observed in corn between levels of ENA and ENB1, ENA and ENB, ENA1 and ENB1, as well as in wheat between BEA and ENA, BEA and ENA1, BEA and ENB, BEA and ENB1, ENA and ENA1, ENA and ENB, ENA and ENB1, ENA1 and ENB, ENA1 and ENB1, and ENB and ENB1. These results demonstrate that co-contamination of BEA and enniatins (ENNs) in corn- and wheat-based samples from Shandong, China is very common. More data on the contamination of five mycotoxins in cereal and cereal-based samples nationwide are needed.

Presented at Indoor Air Seminar (Helsinki, Finland) 15 March, 2018. ISBN 978-952-5236-46-0. Mittasimme aineiden (hometoksiinit; rakennus- ja saneeraustuotteiden kemikaalit, siivousaineet) kulkeutumista päästölähteistä ilmaan tutkimuskammioissa. Tutkittavat aineet annosteltiin kammioiden pohjalla oleviin avoimiin astioihin. Kammioiden ilman kosteutta säädettiin lisäämällä pohjalle vettä ja ilmankostutin- ja kuivainlaitteilla. Lämpötilaa, kosteutta ja TVOC-pitoisuutta monitoroitiin jatkuvatoimisilla antureilla. Kammioilman vesihöyry ja vesiaerosolit tiivistettiin nestemäiseksi vedeksi kuivaimen säiliöön. Kun tiivistevesi analysoitiin, havaittiin, että mikrobitoksiinit (okratoksiini A, enniatiini B) ja siivous- ja saneeraustuotteiden sisältämät antimikrobiset (PHMG) aineet, neutraalit (polyglykoli-alkoksyylit) ja kationiset tensidit (DDDMAC) siirtyivät pinnoilta ilmaan ja sieltä ilmankuivaimen kondenssivesisäiliöön. We measured mobility of nonvolatile fungal toxins and chemical substances in products used in building construction or maintenance into indoor air. The measurement was conducted in enclosed chambers provided with sensors for continuous monitoring of air humidity, temperature, organic substance (TVOC). Airborn humidicity was cyclically up/down adjusted using humidifier / dehumidifier. Substances tested for emission, were placed in open dishes on chamber floor. Airborn humidity was condensed into liquid water, harvested in dehumidifer storage container, and analysed at the end of each experiment.We found that nonvolatile substances such as fungal toxins (ochratoxin A), non-ionic and cationic tensides (polyoxyethylene alkanol ethers, didecyldimethyl ammonium chloride) and antimicrobial biocidal substances (PHMG) , contained in cleaning and sanitation products in buildings, translocated were recovered in the the dehumidifer condensate, thus indicating airborn migration.

Toxicogenic fungal moulds have been previously identified in Irish farm silages and are known to be capable of producing mycotoxins. Mycotoxins are detrimental to animal health and performance and can be transferred through feed to animal produce. The objectives of this study were to identify and quantify the challenge posed to livestock from mycotoxins in Irish silages and determine whether conventional chemical characteristics could be used as indicators of mycotoxin occurrence. Over the 2-year period, 300 silages were sampled on 150 farms and these consisted of round-baled grass silages (n = 115) and pit silages of either grass (n = 175) or maize (n = 10). There was no significant difference in mycotoxin concentrations across silage types in either year, except for baled silage containing higher concentrations of enniatin B compared with pit silage in Year 1. Conventional chemical characteristics of silages were generally not reliable predictors for mycotoxin incidences; however, dry-matter digestibility, crude protein, fermentation products and ash did predict the incidence of enniatins and beauvericin. The incidence and concentration of the twenty measured mycotoxins were generally low and individual mycotoxin concentrations recorded were considerably lower than current EU directive or guidance thresholds. Non-regulated mycotoxins measured were similar to or lower than concentrations reported in the literature. Based on current knowledge and the concentrations of individual mycotoxins detected in this study, the challenge to Irish livestock and livestock products from animal consuming silage is generally low. However, the additive or synergistic effects of multiple mycotoxins in silage are unknown.

In an effort to enforce the knowledge on mycotoxin occurrence and co-occurrence in dates and dried fruits, 228 samples purchased from Tunisian and Spanish markets were subjected to multi-mycotoxin liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis of 16 mycotoxins. At least one mycotoxin was detected in 160 samples (70%). The frequency of contaminated samples was 83%, 80%, 64%, 59% and 26% for dates, dried vine fruits, figs, apricots and plums, respectively but none of the analyzed samples contained detectable levels of fumonisins (FB1, FB2, FB3) or T-2 toxin (T2). The incidence and levels of mycotoxins varied in samples. The most prevalent mycotoxin was enniatin B (EnnB) (54%), followed by enniatinA1 (EnnA1) (36%); aflatoxins (AFs) (23%) and ochratoxin A (OTA) (22%). Thirteen samples contained AFs in levels that exceed the maximum limits established in EU legislation. The simultaneous contamination with 2–6 mycotoxins was observed in 51% of the analyzed samples. Lastly, intakes of the detected mycotoxins were calculated for average adult consumers and compared with the tolerable daily intakes (TDI). Data obtained were used to estimate the potential exposure levels.

The Fusarium mycotoxin enniatin B (ENN B) is a so-called emerging mycotoxin frequently contaminating poultry feed. To investigate the impact of chronic ENN B exposure on animal health, broiler chickens were fed either a diet naturally contaminated with ENN B (2352 µg/kg) or a control diet (135 µg/kg) for 2, 7, 14, or 21 days. ENN B concentrations were determined in plasma and liver using a validated ultra-high performance liquid chromatography—tandem mass spectrometry UHPLC-MS/MS method. Liver was evaluated histologically, and the villus length and crypt depth of the duodenum, jejunum, and ileum were measured. Histopathology of the livers did not reveal major abnormalities. Feeding an ENN B-contaminated diet could possibly inhibit the proliferation of enterocytes in the duodenal crypts, but did not affect villus length, crypt depth, or villus length-crypt depth ratio of the jejunum and ileum. ENN B levels in plasma and liver were significantly higher in the ENN B-fed group and ranged between

Enniatins (Enns) are mycotoxins produced by Fusarium spp. and are widely distributed contaminants of cereals and derivate products. Among the different identified enniatins, Enn B is the most relevant analogue in cereals in Europe. Therefore, the aim of this study was to investigate for the first time the occurrence of Enn B and Enn B phase I metabolites in 300 human urine samples throughout an ultrahigh-performance liquid chromatography-high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) methodology. Three different sample preparation procedures were evaluated and salting-out liquid-liquid extraction showed satisfactory validation results. Enn B was quantified in 83.7% of samples ranging from 0.006 to 0.391 ng/mL (average content: 0.016 ng/mL). In line with the in vitro observations with human liver microsomes, in the here analyzed samples the Enn B monooxygenated, N-demethylated and dioxygenated metabolites were tentatively found in 87.7%, 96.3% and 6.7% of samples. The data of this pilot biomonitoring survey indicate a frequent intake of enniatins in Italy, supporting further toxicological studies to provide better basis for understanding their potential effects in humans.

Beauvericin (BEA) and Enniatins (ENN) are mycotoxins produced by Fusarium fungi detected in food and feed; there are insufficient data to establish their reference values. To evaluate BEA and ENN oral toxicity, an integrated approach was applied. Among ENN, Enniatin B (ENNB) was selected as test substance. The approach is composed by: i) in vitro and acute in vivo genotoxicity tests; ii) a repeated-dose oral toxicity study focused on genotoxic, immune, endocrine, nervous endpoints and the reproductive/developmental toxicity screening. For BEA, all the genotoxicity endpoints yielded negative results excluding Comet assay in duodenum and kidney after repeated doses. BEA immunotoxicity was observed in female mice, concentrated in number and functional activity of effector T cells in the spleen. Based on the repeated-dose BEA study, the No Observed Adverse Effect Level (NOAEL) for female mice is 1 mg/kg b.w. per day (increased thyroid pycnotic nuclei and endometrial hyperplasia). In males, the NOAEL is 0.1 mg/kg b.w. per day (reduced colloid and altered T4 serum levels). Maternal NOAEL is 0.1 mg/kg b.w. per day (increased thymus weight), developmental NOAEL is 10 mg/kg b.w. per day. For ENNB, the results support a genotoxic effect in bone marrow and liver cells after acute treatment, but not after repeated exposure. Immunotoxic ENNB effects were observed in both genders, suggestive of a suppressive/inhibiting activity particularly evident in males. Based on the repeated-dose ENNB study, the NOAEL for females is 0.18 mg/kg b.w. per day (histomorphometrical effects on thymus, uterus and spleen). In male mice, the NOAEL is 1.8 mg/kg b.w. per day (enterocyte vacuolization in duodenum and increased Reactive Oxygen Species and reduced Glutathione brain levels). The maternal NOAEL is 1.8 mg/kg b.w. per day (decreased white pulp area and increased red/white pulp area ratio in spleen), developmental NOAEL is 18 mg/kg b.w. per day.

The waste management occupational environment is recognized by the simultaneous presence of several substances and biologic agents. Therefore, workers are exposed simultaneously to multiple contaminants. Occupational exposure to aflatoxin B1 in one Portuguese waste sorting plant was already reported. However, besides this mycotoxin, data regarding fungal contamination showed that exposure to other mycotoxins could be expected. A study was developed to analyze if exposure to other mycotoxins besides aflatoxin B1 was occurring in the workers from the waste sorting plant previously assessed and to discuss how these findings need to be considered in the risk assessment process. In addition to aflatoxin B1 detected previously by ELISA, two additional mycotoxins and one mycotoxin degradation product were detected and quantified by a multi-mycotoxin HPLC-MS/MS approach: Enniatin B and ochratoxin A as well as 2’R-ochratoxin A. Besides the confirmation of co-exposure to several mycotoxins, results probably indicate different exposure routes for the mycotoxins reported.

Mycotoxin enniatin B (ENN B) is a secondary metabolism product by Fusarium fungi. It is a well-known antibacterial, antihelmintic, antifungal, herbicidal, and insecticidal compound. It has been found as a contaminant in several food commodities, particularly in cereal grains, co-occurring also with other mycotoxins. The primary mechanism of action of ENN B is mainly due to its ionophoric characteristics, but the exact mechanism is still unclear. In the last two decades, it has been a topic of great interest since its potent mammalian cytotoxic activity was demonstrated in several mammalian cell lines. Moreover, the co-exposure in vitro with other mycotoxins enhances its toxic potential through synergic effects, depending on the concentrations tested. Despite its clear cytotoxic effect, European Food Safety Authority stated that acute exposure to ENNs, such as ENN B, does not indicate concern for human health, but a concern might be the chronic exposure. However, given the lack of relevant toxicity data, no firm conclusion could be drawn and a risk assessment was not possible. In fact, very few studies have been carried out in vivo and, in these studies, no adverse effects were observed. So, research on toxicological effects induced by ENN B is still on-going. Recently, some studies are dealing with new advances regarding ENN B. This review summarizes the information on biochemical and biological activity of ENN B, focusing on toxicological aspects and on the latest advances in research on ENN B.

Mycotoxin enniatin B (ENN B) is a secondary metabolism product by Fusarium fungi. It is a well-known antibacterial, antihelmintic, antifungal, herbicidal, and insecticidal compound. It has been found as a contaminant in several food commodities, particularly in cereal grains, co-occurring also with other mycotoxins. The primary mechanism of action of ENN B is mainly due to its ionophoric characteristics, but the exact mechanism is still unclear. In the last two decades, it has been a topic of great interest since its potent mammalian cytotoxic activity was demonstrated in several mammalian cell lines. Moreover, the co-exposure in vitro with other mycotoxins enhances its toxic potential through synergic effects, depending on the concentrations tested. Despite its clear cytotoxic effect, European Food Safety Authority stated that acute exposure to ENNs, such as ENN B, does not indicate concern for human health, but a concern might be the chronic exposure. However, given the lack of relevant toxicity data, no firm conclusion could be drawn and a risk assessment was not possible. In fact, very few studies have been carried out in vivo and, in these studies, no adverse effects were observed. So, research on toxicological effects induced by ENN B is still on-going. Recently, some studies are dealing with new advances regarding ENN B. This review summarizes the information on biochemical and biological activity of ENN B, focusing on toxicological aspects and on the latest advances in research on ENN B.

Background Genome mining approaches predict dozens of biosynthetic gene clusters in each of the filamentous fungal genomes sequenced so far. However, the majority of these gene clusters still remain cryptic because they are not expressed in their natural host. Simultaneous expression of all genes belonging to a biosynthetic pathway in a heterologous host is one approach to activate biosynthetic gene clusters and to screen the metabolites produced for bioactivities. Polycistronic expression of all pathway genes under control of a single and tunable promoter would be the method of choice, as this does not only simplify cloning procedures, but also offers control on timing and strength of expression. However, polycistronic gene expression is a feature not commonly found in eukaryotic host systems, such as Aspergillus niger. Results In this study, we tested the suitability of the viral P2A peptide for co-expression of three genes in A. niger. Two genes descend from Fusarium oxysporum and are essential to produce the secondary metabolite enniatin (esyn1, ekivR). The third gene (luc) encodes the reporter luciferase which was included to study position effects. Expression of the polycistronic gene cassette was put under control of the Tet-On system to ensure tunable gene expression in A. niger. In total, three polycistronic expression cassettes which differed in the position of luc were constructed and targeted to the pyrG locus in A. niger. This allowed direct comparison of the luciferase activity based on the position of the luciferase gene. Doxycycline-mediated induction of the Tet-On expression cassettes resulted in the production of one long polycistronic mRNA as proven by Northern analyses, and ensured comparable production of enniatin in all three strains. Notably, gene position within the polycistronic expression cassette matters, as, luciferase activity was lowest at position one and had a comparable activity at positions two and three. Conclusions The P2A peptide can be used to express at least three genes polycistronically in A. niger. This approach can now be applied to heterologously express entire secondary metabolite gene clusters polycistronically or to co-express any genes of interest in equimolar amounts. Electronic supplementary material The online version of this article (doi:10.1186/s12934-017-0780-z) contains supplementary material, which is available to authorized users.

Emerging Fusarium and Alternaria mycotoxins gain more and more interest due to their frequent contamination of food and feed, although in vivo toxicity and toxicokinetic data are limited. Whereas the Fusarium mycotoxins beauvericin, moniliformin and enniatins particularly contaminate grain and grain-based products, Alternaria mycotoxins are also detected in fruits, vegetables and wines. Although contamination levels are usually low (mu g/kg range), higher contamination levels of enniatins and tenuazonic acid may occasionally occur. In vitro studies suggest genotoxic effects of enniatins A, A1 and B1, beauvericin, moniliformin, alternariol, alternariol monomethyl ether, altertoxins and stemphyltoxin-III. Furthermore, in vitro studies suggest immunomodulating effects of most emerging toxins and a reproductive health hazard of alternariol, beauvericin and enniatin B. More in vivo toxicity data on the individual and combined effects of these contaminants on reproductive and immune system in both humans and animals is needed to update the risk evaluation by the European Food Safety Authority. Taking into account new occurrence data for tenuazonic acid, the complete oral bioavailability, the low total body clearance in pigs and broiler chickens and the limited toxicity data, a health risk cannot be completely excluded. Besides, some less known Alternaria toxins, especially the genotoxic altertoxins and stemphyltoxin III, should be incorporated in risk evaluation as well.

Bipolaris oryzae is a phytopathogenic fungus causing a brown spot disease in rice, and produces substance that strongly perturbs motility and membrane integrities of boar spermatozoa. The substance was isolated from the liquid culture of the fungal strain using extraction and a multi-step semi-preparative HPLC procedures. Based on the results of mass spectrometric and 2D NMR techniques, the bioactive molecule was identified as ophiobolin A, a previously described sesterterpene-type compound. The purified ophiobolin A exhibited strong motility inhibition and viability reduction on boar spermatozoa. Furthermore, it damaged the sperm mitochondria significantly at sublethal concentration by the dissipation of transmembrane potential in the mitochondrial inner membrane, while the plasma membrane permeability barrier remained intact. The study demonstrated that the cytotoxicity of ophiobolin A toward somatic cell lines is higher by 1–2 orders of magnitude compared to other mitochondriotoxic mycotoxins, and towards sperm cells unique by replacing the progressive motility by shivering tail beating at low exposure concentration.

By using extraction experiments and γ -activity measurements, the extraction constant corresponding to the equilibrium Sr 2+ (aq) + 2A − (aq) + 1 (nb) ⇄ 1 · Sr 2+ (nb) + 2A − (nb) occurring in the two-phase water–nitrobenzene system (A − = picrate, 1 = enniatin B; aq = aqueous phase, nb = nitrobenzene phase) was evaluated as log K ex ( 1 · Sr 2+ ,2A − ) = 3.4 ± 0.1. Further, the stability constant of the 1 · Sr 2+ complex in nitrobenzene saturated with water was calculated for a temperature of 25 °C: log β nb ( 1 · Sr 2+ ) = 12.5 ± 0.1. Finally, applying quantum mechanical DFT calculations, the most probable structure of the proved 1 · Sr 2+ cationic complex was derived. In the resulting complex, which is most energetically favored, the “central” cation Sr 2+ is bound by nine bonding interactions to the corresponding nine oxygen atoms of the parent enniatin B ligand. The interaction energy of the considered 1 · Sr 2+ complex was found to be −877.4 kJ/mol, confirming also the formation of this complex.

The contamination levels of beauvericin and four enniatins, A, A1, B and B1, in 207 samples of wheat flour and corn grits on the Japanese market were determined by an analytical method based on LC-MS/MS. The toxins were extracted from samples with acetonitrile–water (85:15, v/v) and then purified with C18 cartridges. The method was validated in a single laboratory using spiked samples at two levels; the recovery of the five toxins ranged from 91.1% to 113.8%. Enniatin B was frequently detected in imported wheat flour (81.8%) and domestic wheat flour (85.6%), and the highest concentration of enniatin B was present in a domestic wheat sample (633 μg kg−1). In corn grits, beauvericin was found in 34% of the samples, but enniatins were not detected at all. The maximum concentration of beauvericin in corn grits was 26.1 μg kg−1. Deoxynivalenol and nivalenol in the same samples were determined by a method using an immunoaffinity column. Co-contamination of deoxynivalenol and enniatins was observed in 61% of the imported wheat samples and in 58% of the domestic wheat samples. These results suggest the need for a risk assessment for cyclic depsipeptide mycotoxins in Japan and a study on the synergistic effect of deoxynivalenol and enniatins.

Introduction - Occupational exposures are characterized by being complex and associated to co-exposure to several contaminants by different exposure routes. Even if exposure occurs to only a chemical agent, it can have different exposure routes and can result in different health effects. The waste management setting is recognized by the presence of several chemical and biologic agents in the workplaces. Recently, it was reported occupational exposure to Aflatoxin B1 (AFB1) in one Portuguese waste management industry. However, data regarding to fungal burden showed that exposure to other mycotoxins should be expected. Aim of the study - The aim of the present work was to know if workers from this waste management industry were exposed to other mycotoxins besides AFB1.

Six new cyclohexadepsipeptides, beauvenniatins A–E ( 1 – 5 ), and beauvericin J ( 6 ), together with the known beauvericin ( 7 ) and enniatin B ( 8 ), were isolated from the fungus Acremonium sp. BCC 28424. The productions of minor derivatives 3 – 6 , possessing an N -methyl- l -tyrosine, were enhanced by feeding l -tyrosine in the liquid fermentation medium. Antimalarial, antitubercular, and cytotoxic activities of these cyclodepsipeptides were evaluated.

A pathogenic fungus, Fusarium sporotrichioides Sherb., was isolated from Hypericum barbatum Jacq. The volatile compounds of broth and mycelium were analyzed using GC–MS and three cyclodidepsipeptides (dioxomorpholines), 3,6-di(propan-2-yl)-4-methyl-morpholine-2,5-dione, 3-(2-methylpropyl)-6-(propan-2-yl)-4-methyl-morpholine-2,5-dione and 3-(butan-2-yl)-6-(propan-2-yl)-4-methyl-morpholine-2,5-dione, were found for the first time in the natural products. The structures of the compounds were confirmed by comparison of the analytical data for the natural products with samples obtained via synthetic methods. The conformational features and vibrational spectra of the three cyclodidepsipeptides were characterized by density functional theory (DFT) calculations and IR spectroscopy. The cyclic hexadepsipeptide enniatin B was identified by a LC–MS/MS analysis of the non-volatile products of broth and mycelium. The above-mentioned three cyclodidepsipeptides are probably synthesized using similar biosynthetic ways to enniatin B involving a nonribosomal mechanism.

A LC-MS/MS method for the detection of beauvericin and the four enniatins A, A1, B, and B1 in maize and maize silage was developed. The method uses direct injection of maize extracts without any tedious and laborious cleanup procedures. The limit of quantification was determined at 13 ng g(-1) for beauvericin and at 17, 34, 24, and 26 ng g(-1) for enniatins A, A1, B, and B1, respectively. The method was used in surveys of the compounds in fresh maize samples collected at harvest in 2005 and 2006. All samples had the same distribution of the enniatins: B > B1 > A1 > A. Enniatin B was present in 90% of the samples in 2005 and in 100% in 2006 at levels up to 489 and 2598 ng g(-1), respectively. Beauvericin contamination was more frequently detected in 2006 than in 2005 (89 and 10%, respectively) and in higher amounts (988 and 71 ng g(-1), respectively). The occurrence of beauvericin and the four enniatins was examined in 3-month-old maize silage stacks from 20 different farms. As observed in fresh maize, enniatin B was the most abundant compound in ensiled maize and was found from 19 stacks at levels up to 218 ng g(-1). The stability of enniatin B in maize silage was assessed by analyzing samples from 10 of the silage stacks taken after 3, 7, and 11 months of ensiling. Enniatin B could be detected at all locations after 11 months and appeared to be stable during ensiling.

Reasons for mammalian cell toxicity observed in barley and spring wheat grains were sought. Streptomyces sp. isolates from wheat and barley produced heat-stable methanol-soluble substances which inhibited the motility of exposed porcine spermatozoa used as a toxicity indicator. Several barley isolates produced antimycin A (2 to 5 ng/mg wet wt of biomass), a macrolide antibiotic known to block oxygen utilization in mitochondria. The antimycinproducing isolates were members of the Streptomyces albidoflavus group. In in vitro assays with porcine kidney tubular epithelial cells, the specific toxicity of antimycin A towards mitochondria was higher than that of the mycotoxin enniatin B but lower than that of the mitochondriotoxins cereulide and paenilide, produced by food-related Bacillus cereus and Paenibacillus tundrae, respectively. The toxic wheat isolates, related to Streptomyces sedi, did not produce antimycin A and or any other known toxin. Our results suggest that the presence of toxin-producing streptomycetes in stored cereal grains may pose a thus far unrecognized threat for food and feed safety.

Enniatin B (ENN B), deoxinivalenol (DON) and alternariol (AOH) are secondary metabolites of filamentous fungi. These mycotoxins are contaminants of vegetables and cereals. They are cytotoxic and their effects are enhanced by their mixtures. The objectives of this study were to compare the cytotoxicity of ENN B, DON and AOH alone or in combination in human adenocarcinoma (Caco-2) cells and to evaluate the type of interactions of mycotoxin mixtures by the isobologram analysis. Cells were treated with concentrations ranging from 1.85 to 90μM (AOH) and from 0.312 to 10μM (for ENN B and DON), individually and in combination of two and three mycotoxins (from 1.85 to 30μM for AOH and from 0.312 to 5μM for ENN B and DON). The relation ratios between the mixtures DON+ENN B was 1:1; AOH+DON and ENN B+AOH was 1:6, and for the tertiary combination DON, ENN B and AOH 1:1:6. The IC50 value of ENN B and DON were 3.87 and 5.54μM, respectively. No IC50 values were obtained for the AOH at any time tested in Caco-2 cells. With the isobologram the type of interaction between mycotoxin was evaluated. Synergistic, antagonistic and addictive effect was observed for the combination studied depending on the concentration affected. Mycotoxins combinations reduce cellular viability in the following increasing order: (DON+ENN B)>(ENN B+AOH)>(DON+AOH)>(DON+AOH+ENN B).

A nine-step (longest linear) batch total synthesis of the cyclic hexadepsipeptide (−)-enniatin B is described. The synthesis minimizes precipitation during reaction conditions for adaptability to flow synthesis. The route was used to prepare >100 mg of the natural product.

The crystal structure of the complex of enniatin B, cyclo[-(L-MeVal-D-Hyi)3-](C33H57N3O9), with KNCS is determined by X-ray diffraction [CuKαradiation, R = 0.0594 for 7925 reflections with I > 2 σ(I)]. The crystals belong to the space group P3, a = 24.448(5) A, c = 23.578(5) A, V = 12277(9) A3, and Z = 6. The unit cell contains 12 symmetrically independent molecules of the antibiotic, which are located on crystallographic threefold axes. The K+ ions are located on the threefold axes and are coordinated by the carbonyl oxygen atoms of the hydroxy acid or amino acid residues of the enniatin molecules to form prisms, twisted prisms, and antiprisms. All the independent enniatin molecules retain the principal conformational features revealed earlier in the structures of enniatin B and its complexes.

We evaluated Fusarium contamination and the levels of hexadepsipeptide mycotoxins in 13 wheat samples affected by head blight in Finland. Fusarium avenaceum was the dominant species (91%) isolated from all samples, but isolates of F. culmorum (4%), F. tricinctum (3%), and F. poae (2%) also were recovered. Beauvericin (0.64 to 3.5 μg/g) was detected in all 13 samples. Enniatin B (trace to 4.8 μg/g) was detected in 12 samples, enniatin B 1 (trace to 1.9 μg/g) was detected in 8 samples, and enniatin A 1 (trace to 6.9 μg/g) was detected in 10 samples. Ten of 13 strains of F. avenaceum and 2 strains of F. poae and F. tricinctum produced beauvericin in culture on rice (trace to 70, 9.4, and 33 μg/g, respectively). All strains also produced enniatins (trace to 2,700 μg/g). This is the first report on the natural cooccurence of beauvericin and enniatins in wheat infected predominantly by F. avenaceum .

Twenty-nine Fusarium isolates, representing three new taxa originated by Nirenberg from F. sambucinum Fuckel sensu lato, namely: F. sambucinum Fuckel sensu stricto, F. venenotum Nirenb., and F. torulosum (Berk.Curt.) Nirenb., were tested for in vitro production of toxic secondary metabolites on autoclaved corn kernels. F. sambucinum sensu stricto was able to produce type A trichothecenes and enniatin B (EB). In particular, amongst the 14 isolates tested, 5 produced only diacetoxyscirpenol (DAS) (up to 700 micrograms/g); 1 produced only neosolaniol (NEOS) (250 micrograms/g); 2 produced T-2 toxin (T-2) + NEOS (up to 175 and 150 micrograms/g, respectively); 1 produced NEOS + DAS (300 and 100 micrograms/g, respectively); and 5 produced DAS + EB (up to 500 and 140 micrograms/g, respectively). All six isolates of F. venenotum were able to produce only DAS (up to 100 micrograms/g). F. torulosum produced no trichothecenes, but four out of nine tested isolates were able to produce EB (up to 140 micrograms/g). Zearalenones and type B trichothecenes were not found. The toxicity of the culture extracts towards Artemia salina L. was correlated in general with the occurrence of the above toxins, except for some F. torulosum strains. However, the lack of correlation between the amounts of toxins recovered and toxic activity observed in the Geotrichum candidum Link ex Pers. and A. salina assays suggested the presence of unknown toxic compounds.