Sample records for deoxynivalenol don vomitoxin
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Nontransmission of deoxynivalenol (vomitoxin) to milk following oral administration to dairy cows.
Prelusky, D B; Trenholm, H L; Lawrence, G A; Scott, P M
1984-10-01
The absorption of deoxynivalenol (DON; vomitoxin), a trichothecene mycotoxin produced by Fusarium species, was studied in the dairy cow. Serum and milk DON levels were quantitated following a single oral dose of 920 mg DON to each of two lactating cows of similar weight. Maximum blood levels for the two animals following DON administration were 200 and 90 ng/ml serum, occurring at times 4.7 and 3.5 hr, respectively. By 24 hr after dosing only trace levels (less than 2 ng/ml) were still detectable. DON in its conjugated form accounted for 24-46% of the total levels present in serum. Free and conjugated DON were also present in cow's milk, but only extremely low amounts (less than 4 ng/ml) were detected. Detection of DON was carried out utilizing Sep-Pak C18 extraction cartridges for isolation, with additional purification of the sample achieved by passing the extract through a short charcoal/alumina column. The extract was then reacted with N-heptafluorobutyrylimidazole prior to quantitation of the resulting DON-tris-heptafluorobutyrate derivative by combined gas chromatography-quadrupole mass spectrometry, using multiple selected ion monitoring. Detection limits were as low as 1 ng/ml (1 ppb).
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Overt signs of toxicity to dogs and cats of dietary deoxynivalenol.
Hughes, D M; Gahl, M J; Graham, C H; Grieb, S L
1999-03-01
Studies were conducted to determine the dietary amounts of deoxynivalenol (DON; vomitoxin) in dog and cat food that are required to produce overt signs of toxicity (e.g., vomiting or reduced food intake). Wheat naturally contaminated with 37 mg of DON/kg was used to manufacture pet foods containing 0, 1, 2, 4, 6, 8, and 10 mg of DON/kg. Deoxynivalenol concentration in pet food following manufacture was unchanged, indicating that the toxin was stable during conventional extrusion processing. Dogs previously fed DON-contaminated food were able to preferentially select uncontaminated food. Dogs not previously exposed to DON-contaminated food consumed equal quantities of contaminated and uncontaminated food. There was no effect of 6 mg of DON/kg on dog food digestibility. Food intake of dogs was significantly reduced by DON concentrations greater than 4.5 +/- 1.7 mg/kg, and DON greater than 7.7 +/- 1.1 mg/kg reduced cat food intake. Vomiting by dogs and cats was commonly observed at the 8 and 10 mg DON levels.
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Pestka, James J.
2013-01-01
Deoxynivalenol (DON, vomitoxin), a trichothecene mycotoxin produced by Fusarium sp. that frequently occurs in cereal grains, has been associated with human and animal food poisoning. Although a common hallmark of DON-induced toxicity is the rapid onset of emesis, the mechanisms for this adverse effect are not fully understood. Recently, our laboratory has demonstrated that the mink (Neovison vison) is a suitable small animal model for investigating trichothecene-induced emesis. The goal of this study was to use this model to determine the roles of two gut satiety hormones, peptide YY3–36 (PYY3–36) and cholecystokinin (CCK), and the neurotransmitter 5-hydroxytryptamine (5-HT) in DON-induced emesis. Following ip exposure to DON at 0.1 and 0.25mg/kg bw, emesis induction ensued within 15–30min and then persisted up to 120min. Plasma DON measurement revealed that this emesis period correlated with the rapid distribution and clearance of the toxin. Significant elevations in both plasma PYY3–36 (30–60min) and 5-HT (60min) but not CCK were observed during emesis. Pretreatment with the neuropeptide Y2 receptor antagonist JNJ-31020028 attenuated DON- and PYY-induced emesis, whereas the CCK1 receptor antagonist devezapide did not alter DON’s emetic effects. The 5-HT3 receptor antagonist granisetron completely suppressed induction of vomiting by DON and the 5-HT inducer cisplatin. Granisetron pretreatment also partially blocked PYY3–36-induced emesis, suggesting a potential upstream role for this gut satiety hormone in 5-HT release. Taken together, the results suggest that both PYY3–36 and 5-HT play contributory roles in DON-induced emesis. PMID:23457120
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Effects of Bread Making and Wheat Germ Addition on the Natural Deoxynivalenol Content in Bread
Giménez, Isabel; Blesa, Jesús; Herrera, Marta; Ariño, Agustín
2014-01-01
Deoxynivalenol (DON, vomitoxin) is a type-B trichothecene mycotoxin produced by several field fungi such as Fusarium graminearum and Fusarium culmorum and known to have various toxic effects. This study investigated the effect of the bread making process on the stability of DON in common bread and wheat germ-enriched bread using naturally contaminated ingredients at the level of 560 µg/kg. The concentration of DON and its evolution during bread making were determined by immunoaffinity column cleanup followed by liquid chromatography with diode array detection (HPLC-DAD). During the bread making process, DON was reduced by 2.1% after fermentation and dropped by 7.1% after baking, reaching a maximum reduction of 19.8% in the crust as compared with a decrease of 5.6% in the crumb. The addition of 15% wheat germ to the dough did not affect DON stability during bread making, showing an apparent increase of 3.5% after fermentation and a reduction by 10.2% after baking. PMID:24451845
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Abouzied, M M; Azcona, J I; Braselton, W E; Pestka, J J
1991-01-01
To assess the potential for mycotoxin contamination of the human food supply following the 1988 U.S. drought, 92 grain food samples were purchased from retail outlets in the summer of 1989 and surveyed for aflatoxin B1, zearalenone, and deoxynivalenol (DON [vomitoxin]) by monoclonal antibody-based competitive enzyme-linked immunosorbent assay (ELISA). Only one sample (buckwheat flour) was found to contain aflatoxin B1 (12 ng/g), whereas zearalenone was found in 26% of the samples at a mean concentration of 19 ng/g. In contrast, the DON ELISA was positive in 50% of the samples at a detection level of 1.0 micrograms/g. Between 63 and 88% of corn cereals, wheat flour/muffin mixes, rice cereals, and corn meal/muffin mixes yielded positive results for DON, whereas 25 to 50% of oat cereals, wheat- and oat-based cookies/crackers, corn chips, popcorn, and mixed-grain cereals were positive for DON. The mean DON content of the positive samples was 4.0 micrograms/g, and the minimum and maximum levels were 1.2 and 19 micrograms/g, respectively. When positive ELISA samples were also analyzed by high-performance liquid chromatography, a strong correlation between the two methods was found. The presence of DON in the two highest samples, corn meal and mixed-grain cereal, which contained 19 and 16 micrograms/g, respectively, was quantitatively confirmed by gas chromatography-mass spectrometry. The results indicated that DON was present in 1989 retail food products at concentrations that exceeded those found in previous market surveys and that have been experimentally associated with impaired animal health. Images PMID:1828138
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Casale, W.L.
1987-01-01
The 12,13-epoxytrichothecene mycotoxins deoxynivalenol (DON, vomitoxin) and T-2 toxin inhibited protein synthesis in vivo and in cell-free systems from wheat and maize, host plants of trichothecene-producing Fusarium spp.Protein synthesis in tissue (leaf discs and kernel sections) was measured by incorporation of /sup 3/H-leucine into acetone:ethanol insoluble material, and in cell-free translation systems from wheat embryos and maize seedling plumules by incorporation of /sup 3/H-leucine into trichloroacetic acid-insoluble material. The toxin concentration inhibiting 50% of /sup 3/H-leucine incorporation (ID/sub 50/) by several maize varieties were 0.9 ..mu..M (T-2 toxin) and 9-22 ..mu..M (DON). ID/sub 50/ values for wheat were 0.25 ..mu..Mmore » (T-2 toxin) and 4.5 ..mu..M (DON).« less
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Clark, Erica S.; Flannery, Brenna M.; Gardner, Elizabeth M.; Pestka, James J.
2015-01-01
Deoxynivalenol (DON), a trichothecene mycotoxin that commonly contaminates cereal grains, is a public health concern because of its adverse effects on the gastrointestinal and immune systems. The objective of this study was to compare effects of DON on anorectic responses in aged (22 mos) and adult (3 mos) mice. Aged mice showed increased feed refusal with both acute i.p. (1 mg/kg and 5 mg/kg) and dietary (1, 2.5, 10 ppm) DON exposure in comparison to adult mice. In addition to greater suppression of food intake from dietary DON exposure, aged mice also exhibited greater but transient body weight suppression. When aged mice were acutely exposed to 1 mg/kg bw DON i.p., aged mice displayed elevated DON and DON3GlcA tissue levels and delayed clearance in comparison with adult mice. Acute DON exposure also elicited higher proinflammatory cytokine and satiety hormone responses in the plasma of the aged group compared with the adult group. Increased susceptibility to DON-induced anorexia in aged mice relative to adult mice suggests that advanced life stage could be a critical component in accurate human risk assessments for DON and other trichothecenes. PMID:26492270
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Clark, Erica S; Flannery, Brenna M; Gardner, Elizabeth M; Pestka, James J
2015-10-19
Deoxynivalenol (DON), a trichothecene mycotoxin that commonly contaminates cereal grains, is a public health concern because of its adverse effects on the gastrointestinal and immune systems. The objective of this study was to compare effects of DON on anorectic responses in aged (22 mos) and adult (3 mos) mice. Aged mice showed increased feed refusal with both acute i.p. (1 mg/kg and 5 mg/kg) and dietary (1, 2.5, 10 ppm) DON exposure in comparison to adult mice. In addition to greater suppression of food intake from dietary DON exposure, aged mice also exhibited greater but transient body weight suppression. When aged mice were acutely exposed to 1 mg/kg bw DON i.p., aged mice displayed elevated DON and DON3GlcA tissue levels and delayed clearance in comparison with adult mice. Acute DON exposure also elicited higher proinflammatory cytokine and satiety hormone responses in the plasma of the aged group compared with the adult group. Increased susceptibility to DON-induced anorexia in aged mice relative to adult mice suggests that advanced life stage could be a critical component in accurate human risk assessments for DON and other trichothecenes.
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Tran, S T; Smith, T K
2014-02-01
Deoxynivalenol (DON, vomitoxin) is a trichothecene mycotoxin which can be considered to be an indicator of Fusarium mycotoxin contamination in grain, feed and food. Recent studies have described the presence of glucose conjugated DON, which is a product of plant metabolism, but there is a lack of information available on DON conjugation by fungi. The aim of the current study was, therefore, to investigate the ability of fungi to metabolize DON into hydrolysable conjugated DON. Alternaria alternata (54028 NRRL) and Rhizopus microsporus var. rhizopodiformis (54029 NRRL) were found to be capable of metabolizing DON into hydrolysable conjugated DON. This ranged from 13-23 % conjugation of DON in potato dextrose agar media and from 11-36 % in corn-based media. There was, however, considerable variation between fungal strains in the ability to conjugate DON as only a slight increase in hydrolysable conjugated DON (1-6 %) was observed when incubating with A. oryzae (5509 NRRL). A. oryzae (5509 NRRL) was also shown to degrade DON (up to 92 %) over 21 days of incubation on corn-based media. The current study shows that conjugation of DON can be achieved through fungal metabolism in addition to being a product of plant metabolism.
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Wu, Wenda; Zhou, Hui-Ren; Bursian, Steven J.; Pan, Xiao; Link, Jane E.; Berthiller, Franz; Adam, Gerhard; Krantis, Anthony; Durst, Tony; Pestka, James J.
2014-01-01
The mycotoxin deoxynivalenol (DON) elicits robust anorectic and emetic effects in several animal species. However, less is known about the potential for naturally occurring and synthetic congeners of this trichothecene to cause analogous responses. Here we tested the hypothesis that alterations in DON structure found in the plant metabolite deoxynivalenol-3-glucoside (D3G) and two pharmacologically active synthetic DON derivatives, EN139528 and EN139544, differentially impact their potential to evoke food refusal and emesis. In a nocturnal mouse food consumption model, oral administration with DON, D3G, EN139528, or EN139544 at doses from 2.5 to 10 mg/kg BW induced anorectic responses that lasted up to 16, 6, 6, and 3 h, respectively. Anorectic potency rank orders were EN139544>DON>EN139528>D3G from 0 to 0.5 h but DON>D3G>EN139528>EN139544 from 0 to 3 h. Oral exposure to each of the four compounds at a common dose (2.5 mg/kg BW) stimulated plasma elevations of the gut satiety peptides cholecystokinin and to a lesser extent, peptide YY3–36 that corresponded to reduced food consumption. In a mink emesis model, oral administration of increasing doses of the congeners differentially induced emesis, causing marked decreases in latency to emesis with corresponding increases in both the duration and number of emetic events. The minimum emetic doses for DON, EN139528, D3G, and EN139544 were 0.05, 0.5, 2, and 5 mg/kg BW, respectively. Taken together, the results suggest that although all three DON congeners elicited anorectic responses that mimicked DON over a narrow dose range, they were markedly less potent than the parent mycotoxin at inducing emesis. PMID:25173790
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Wu, Wenda; Zhang, Haibin
2014-01-01
The trichothecene deoxynivalenol (DON), a foodborne mycotoxin found in grain-based foods, has been associated with human and animal food poisoning. Although induction of anorexia has been described as a hallmark of DON-induced toxicity in many animal species, the mechanistic basis for this adverse effect is not fully understood. The purpose of this research was to determine the role of two proinflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in DON-induced anorexia. In a nocturnal mouse food consumption model, DON-induced anorectic response occurred at 1 hr and lasted up to 6 hr. Similar anorectic effects were observed following acute administration of exogenous TNF-α and IL-1β. Oral exposure to DON at 5 mg/kg bw stimulated splenic and hepatic mRNA and plasma protein elevations of TNF-α and IL-1β that corresponded to anorexia induction. Pretreatment with the TNF-α receptor (TNFR) antagonist R-7050 dose-dependently attenuated both TNF-α- and DON-induced anorexia. While, the type 1 IL-1 receptor (IL-1R1) antagonist IL-1RA dose-dependently attenuated both IL-1β- and DON-induced anorexia. Taken together, the results suggest that both TNF-α and IL-1β play contributory role in anorexia induction following oral exposure to DON.
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Park, Juhee; Chang, Hansub; Kim, Dongho; Chung, Soohyun; Lee, Chan
2018-03-16
The Fusarium fungi produce toxic substances called mycotoxins, which can cause disease and harmful effects in grains, livestock, and humans. Deoxynivalenol (DON), also known as vomitoxin, is one of the Fusarium mycotoxins that is known to cause vomiting in livestock. This study shows the occurrence of deoxynivalenol in feedstuffs (compound feed and feed ingredients) between 2009 and 2016 in South Korea. A total of 653 domestic samples were collected at five time points, including 494 compound feed samples and 159 feed ingredient samples. DON contamination levels were analyzed using high-performance liquid chromatography (HPLC) with pretreatment using an immunoaffinity column (IAC). The limit of detection (LOD) and the limit of quantification (LOQ) were estimated at 1-10 µg/kg and 3-35 µg/kg, respectively. Two compound feeds (two gestating sow feed samples) out of 160 pig feed samples exceeded the European Commission (EC) guidance value, while no feed ingredient samples exceeded the EC or South Korean guidance values. There were statistically significant differences in the mean contamination levels of compound feed and feed ingredients that indicated a decreasing trend over time.
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Park, Juhee; Chang, Hansub; Kim, Dongho; Chung, Soohyun
2018-01-01
The Fusarium fungi produce toxic substances called mycotoxins, which can cause disease and harmful effects in grains, livestock, and humans. Deoxynivalenol (DON), also known as vomitoxin, is one of the Fusarium mycotoxins that is known to cause vomiting in livestock. This study shows the occurrence of deoxynivalenol in feedstuffs (compound feed and feed ingredients) between 2009 and 2016 in South Korea. A total of 653 domestic samples were collected at five time points, including 494 compound feed samples and 159 feed ingredient samples. DON contamination levels were analyzed using high-performance liquid chromatography (HPLC) with pretreatment using an immunoaffinity column (IAC). The limit of detection (LOD) and the limit of quantification (LOQ) were estimated at 1–10 µg/kg and 3–35 µg/kg, respectively. Two compound feeds (two gestating sow feed samples) out of 160 pig feed samples exceeded the European Commission (EC) guidance value, while no feed ingredient samples exceeded the EC or South Korean guidance values. There were statistically significant differences in the mean contamination levels of compound feed and feed ingredients that indicated a decreasing trend over time. PMID:29547546
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Jia, Hui; Wu, Wen-Da; Lu, Xi; Zhang, Jie; He, Cheng-Hua; Zhang, Hai-Bin
2017-09-01
Deoxynivalenol (DON), which is a Type B trichothecene mycotoxin produced by Fusarium, frequently contaminates cereal staples, such as wheat, barley and corn. DON threatens animal and human health by suppressing food intake and impairing growth. While anorexia induction in mice exposed to DON has been linked to the elevation of the satiety hormones cholecystokinin and peptide YY3-36 in plasma, the effects of DON on the release of other satiety hormones, such as glucagon-like peptide-1 (GLP-1) and gastric inhibitory peptide (GIP), have not been established. The purpose of this study was to determine the roles of GLP-1 and GIP in DON-induced anorexia. In a nocturnal mouse food consumption model, the elevation of plasma GLP-1 and GIP concentrations markedly corresponded to anorexia induction by DON. Pretreatment with the GLP-1 receptor antagonist Exendin9-39 induced a dose-dependent attenuation of both GLP-1- and DON-induced anorexia. In contrast, the GIP receptor antagonist Pro3GIP induced a dose-dependent attenuation of both GIP- and DON-induced anorexia. Taken together, these results suggest that GLP-1 and GIP play instrumental roles in anorexia induction following oral exposure to DON, and the effect of GLP-1 is more potent and long-acting than that of GIP. © The Author 2017. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.
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Flannery, Brenna M; Clark, Erica S; Pestka, James J
2012-12-01
Consumption of deoxynivalenol (DON), a trichothecene mycotoxin known to commonly contaminate grain-based foods, suppresses growth of experimental animals, thus raising concerns over its potential to adversely affect young children. Although this growth impairment is believed to result from anorexia, the initiating mechanisms for appetite suppression remain unknown. Here, we tested the hypothesis that DON induces the release of satiety hormones and that this response corresponds to the toxin's anorectic action. Acute ip exposure to DON had no effect on plasma glucagon-like peptide-1, leptin, amylin, pancreatic polypeptide, gastric inhibitory peptide, or ghrelin; however, the toxin was found to robustly elevate peptide YY (PYY) and cholecystokinin (CCK). Specifically, ip exposure to DON at 1 and 5mg/kg bw induced PYY by up to 2.5-fold and CCK by up to 4.1-fold. These responses peaked within 15-120 min and lasted up to 120 min (CCK) and 240 min (PPY), corresponding with depressed rates of food intake. Direct administration of exogenous PYY or CCK similarly caused reduced food intake. Food intake experiments using the NPY2 receptor antagonist BIIE0246 and the CCK1A receptor antagonist devazepide, individually, suggested that PYY mediated DON-induced anorexia but CCK did not. Orolingual exposure to DON induced plasma PYY and CCK elevation and anorexia comparable with that observed for ip exposure. Taken together, these findings suggest that PYY might be one critical mediator of DON-induced anorexia and, ultimately, growth suppression.
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Savard, Christian; Pinilla, Vicente; Provost, Chantale; Gagnon, Carl A; Chorfi, Younes
2014-12-05
Deoxynivalenol (DON), also known as vomitoxin, is the most prevalent type B trichothecene mycotoxin worldwide. Pigs show a great sensitivity to DON, and because of the high proportion of grains in their diets, they are frequently exposed to this mycotoxin. The objective of this study was to determine the impact of DON naturally contaminated feed on porcine reproductive and respiratory syndrome virus (PRRSV) infection, the most important porcine viral pathogen in swine. Experimental infections were performed with 30 animals. Piglets were randomly divided into three groups of 10 animals based on DON content of diets (0, 2.5 and 3.5 mg/kg DON). All experimental groups were further divided into subgroups of 6 pigs and were inoculated with PRRSV. The remaining pigs (control) were sham-inoculated with PBS. Pigs were daily monitored for temperature, weight and clinical signs for 21 days. Blood samples were collected and tested for PRRSV RNA and for virus specific antibodies. Results of PRRSV infection showed that ingestion of diet highly contaminated with DON greatly increases the effect of PRRSV infection on weight gain, lung lesions and mortality, without increasing significantly viral replication, for which the tendency is rather directed toward a decrease of replication. These results suggest that PRRSV infection could exacerbate anorectic effect of DON, when ingested in large doses. Results also demonstrate a DON negative effect on PRRSV-specific humoral responses. This study demonstrate that high concentrations of DON naturally contaminated feed decreased the immune response against PRRSV and influenced the course of PRRSV infection in pigs. Copyright © 2014 Elsevier B.V. All rights reserved.
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Pestka, James J.
2012-01-01
Consumption of deoxynivalenol (DON), a trichothecene mycotoxin known to commonly contaminate grain-based foods, suppresses growth of experimental animals, thus raising concerns over its potential to adversely affect young children. Although this growth impairment is believed to result from anorexia, the initiating mechanisms for appetite suppression remain unknown. Here, we tested the hypothesis that DON induces the release of satiety hormones and that this response corresponds to the toxin’s anorectic action. Acute ip exposure to DON had no effect on plasma glucagon-like peptide-1, leptin, amylin, pancreatic polypeptide, gastric inhibitory peptide, or ghrelin; however, the toxin was found to robustly elevate peptide YY (PYY) and cholecystokinin (CCK). Specifically, ip exposure to DON at 1 and 5mg/kg bw induced PYY by up to 2.5-fold and CCK by up to 4.1-fold. These responses peaked within 15–120min and lasted up to 120min (CCK) and 240min (PPY), corresponding with depressed rates of food intake. Direct administration of exogenous PYY or CCK similarly caused reduced food intake. Food intake experiments using the NPY2 receptor antagonist BIIE0246 and the CCK1A receptor antagonist devazepide, individually, suggested that PYY mediated DON-induced anorexia but CCK did not. Orolingual exposure to DON induced plasma PYY and CCK elevation and anorexia comparable with that observed for ip exposure. Taken together, these findings suggest that PYY might be one critical mediator of DON-induced anorexia and, ultimately, growth suppression. PMID:22903826
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Bao, Lei; Oles, Carolyn J; White, Kevin D; Sapp, Chelsea; Trucksess, Mary W
2011-01-01
Deoxynivalenol (DON), also known as vomitoxin, belongs to a class of naturally occurring mycotoxins produced by Fusarium spp. DON, 12, 13-epoxy-3,7 trihydroxytrichothec-9-en-8-one, is one of the most frequently detected mycotoxins in agricultural commodities worldwide. A method consisting of extraction, filtration, column cleanup, and RP-HPLC-UV separation and quantitation was validated for the determination of DON in grains (rice and barley), grain products (whole wheat flour, white flour, wheat germ, and wheat bran), and processed foods (bread, breakfast cereals, and pretzels). A 25 g test portion was extracted with 100 mL acetonitrile-water (84 + 16, v/v). After blending for 3 min, the supernatant was applied to a multifunctional column (MycoSep 225). The purified filtrate (2 mL) was evaporated to dryness and redissolved in the mobile phase. The toxins were then subjected to RP-HPLC-UV analysis. The accuracy and repeatability characteristics of the method were determined. Recoveries of DON added at levels ranging from 0.5 to 1.5 microg/g for all test matrixes were from 75 to 98%. SD and RSD(r) ranged from 0.7 to 11.6% and 0.9 to 12.7%, respectively. Within-laboratory HorRat values were from 0.1 to 0.7 for all matrixes analyzed. The method was found to meet AOAC method performance criteria for grains, grain products, and processed foods. The identity of DON in naturally contaminated test sample extracts was confirmed by HPLC/MS/MS analysis.
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Pestka, James J.
2013-01-01
Although the acute toxic effects of trichothecene mycotoxin deoxynivalenol (DON or vomitoxin), a known cause of human food poisoning, have been well characterized in several animal species, much less is known about closely related 8-ketotrichothecenes that similarly occur in cereal grains colonized by toxigenic fusaria. To address this, we compared potencies of DON, 15-acetyldeoxynivalenol (15-ADON), 3-acetyldeoxynivalenol (3-ADON), fusarenon X (FX), and nivalenol (NIV) in the mink emesis model following intraperitoneal (ip) and oral administration. All five congeners dose-dependently induced emesis by both administration methods. With increasing doses, there were marked decreases in latency to emesis with corresponding increases in emesis duration and number of emetic events. The effective doses resulting in emetic events in 50% of the animals for ip exposure to DON, 15-ADON, 3-ADON, FX, and NIV were 80, 170, 180, 70, and 60 µg/kg bw, respectively, and for oral exposure, they were 30, 40, 290, 30, and 250 µg/kg bw, respectively. The emetic potency of DON determined here was comparable to that reported in analogous studies conducted in pigs and dogs, suggesting that the mink is a suitable small animal model for investigating acute trichothecene toxicity. The use of a mouse pica model, based on the consumption of kaolin, was also evaluated as a possible surrogate for studying emesis but was found unsuitable. From a public health perspective, comparative emetic potency data derived from small animal models such as the mink should be useful for establishing toxic equivalency factors for DON and other trichothecenes. PMID:22997060
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Katika, Madhumohan R.; Department of Health Risk Analysis and Toxicology, Maastricht University; Netherlands Toxicogenomics Centre
Deoxynivalenol (DON) or vomitoxin is a commonly encountered type-B trichothecene mycotoxin, produced by Fusarium species predominantly found in cereals and grains. DON is known to exert toxic effects on the gastrointestinal, reproductive and neuroendocrine systems, and particularly on the immune system. Depending on dose and exposure time, it can either stimulate or suppress immune function. The main objective of this study was to obtain a deeper insight into DON-induced effects on lymphoid cells. For this, we exposed the human T-lymphocyte cell line Jurkat and human peripheral blood mononuclear cells (PBMCs) to various concentrations of DON for various times and examinedmore » gene expression changes by DNA microarray analysis. Jurkat cells were exposed to 0.25 and 0.5 μM DON for 3, 6 and 24 h. Biological interpretation of the microarray data indicated that DON affects various processes in these cells: It upregulates genes involved in ribosome structure and function, RNA/protein synthesis and processing, endoplasmic reticulum (ER) stress, calcium-mediated signaling, mitochondrial function, oxidative stress, the NFAT and NF-κB/TNF-α pathways, T cell activation and apoptosis. The effects of DON on the expression of genes involved in ER stress, NFAT activation and apoptosis were confirmed by qRT-PCR. Other biochemical experiments confirmed that DON activates calcium-dependent proteins such as calcineurin and M-calpain that are known to be involved in T cell activation and apoptosis. Induction of T cell activation was also confirmed by demonstrating that DON activates NFATC1 and induces its translocation from the cytoplasm to the nucleus. For the gene expression profiling of PBMCs, cells were exposed to 2 and 4 μM DON for 6 and 24 h. Comparison of the Jurkat microarray data with those obtained with PBMCs showed that most of the processes affected by DON in the Jurkat cell line were also affected in the PBMCs. -- Highlights: ► The human T cell line Jurkat and human PBMCs were exposed to DON. ► Whole-genome microarray experiments were performed. ► Microarray data indicates that DON affects ribosome and RNA/protein synthesis. ► DON treatment induces ER stress, calcium mediated signaling, NFAT and NF-κB. ► Exposure to DON induces T cell activation, oxidative stress and apoptosis.« less
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Lutz, Matthias P.; Feichtinger, Georg; Défago, Geneviève; Duffy, Brion
2003-01-01
Mycotoxin contamination associated with head blight of wheat and other grains caused by Fusarium culmorum and F. graminearum is a chronic threat to crop, human, and animal health throughout the world. One of the most important toxins in terms of human exposure is deoxynivalenol (DON) (formerly called vomitoxin), an inhibitor of protein synthesis with a broad spectrum of toxigenicity against animals. Certain Fusarium toxins have additional antimicrobial activity, and the phytotoxin fusaric acid has recently been shown to modulate fungus-bacterium interactions that affect plant health (Duffy and Défago, Phytopathology 87:1250-1257, 1997). The potential impact of DON on Fusarium competition with other microorganisms has not been described previously. Any competitive advantage conferred by DON would complicate efforts to control Fusarium during its saprophytic growth on crop residues that are left after harvest and constitute the primary inoculum reservoir for outbreaks in subsequent plantings. We examined the effect of the DON mycotoxin on ecological interactions between pathogenic Fusarium and Trichoderma atroviride strain P1, a competitor fungus with biocontrol activity against a wide range of plant diseases. Expression of the Trichoderma chitinase genes, ech42 and nag1, which contribute to biocontrol activity, was monitored in vitro and on crop residues of two maize cultivars by using goxA reporter gene fusions. We found that DON-producing F. culmorum and F. graminearum strains repressed expression of nag1-gox. DON-negative wild-type Fusarium strains and a DON-negative mutant with an insertional disruption in the tricothecene biosynthetic gene, tri5, had no effect on antagonist gene expression. The role of DON as the principal repressor above other pathogen factors was confirmed. Exposure of Trichoderma to synthetic DON or to a non-DON-producing Fusarium mutant resulted in the same level of nag1-gox repression as the level observed with DON-producing Fusarium. DON repression was specific for nag1-gox and had no effect, either positive or negative, on expression of another key chitinase gene, ech42. This is the first demonstration that a target pathogen down-regulates genes in a fungal biocontrol agent, and our results provide evidence that mycotoxins have a novel ecological function as factors in Fusarium competitiveness. PMID:12788701
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USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON) is a mycotoxin found in wheat that is infected with Fusarium fungus. DON may also be converted to a type of "masked mycotoxin," named deoxynivalenol-3-glucoside (D3G), as a result of detoxification process of the plant. Both DON and D3G are known to be toxic. Due to the lack o...
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Urinary analysis reveals high deoxynivalenol exposure in pregnant women from Croatia.
Sarkanj, Bojan; Warth, Benedikt; Uhlig, Silvio; Abia, Wilfred A; Sulyok, Michael; Klapec, Tomislav; Krska, Rudolf; Banjari, Ines
2013-12-01
In this pilot survey the levels of various mycotoxin biomarkers were determined in third trimester pregnant women from eastern Croatia. First void urine samples were collected and analysed using a "dilute and shoot" LC-ESI-MS/MS multi biomarker method. Deoxynivalenol (DON) and its metabolites: deoxynivalenol-15-glucuronide and deoxynivalenol-3-glucuronide were detected in 97.5% of the studied samples, partly at exceptionally high levels, while ochratoxin A was found in 10% of the samples. DON exposure was primarily reflected by the presence of deoxynivalenol-15-glucuronide with a mean concentration of 120 μg L(-1), while free DON was detected with a mean concentration of 18.3 μg L(-1). Several highly contaminated urine samples contained a third DON conjugate, tentatively identified as deoxynivalenol-7-glucuronide by MS/MS scans. The levels of urinary DON and its metabolites measured in this study are the highest ever reported, and 48% of subjects were estimated to exceed the provisional maximum tolerable daily intake (1 μg kg(-1) b.w.). Copyright © 2013 Elsevier Ltd. All rights reserved.
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Vidal, Arnau; Sanchis, Vicente; Ramos, Antonio J; Marín, Sonia
2015-07-01
The stability of deoxynivalenol (DON), deoxynivalenol-3-glucoside (DON-3-glucoside), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), de-epoxy-deoxynivalenol (DOM-1) and ochratoxin A (OTA) during thermal processing has been studied. Baking temperature, time and initial mycotoxin concentration in the raw materials were assayed as factors. An improved UPLC-MS/MS method to detect DON, DON-3-glucoside, 3-ADON, 15-ADON and DOM-1 in wheat baked products was developed in the present assay. The results highlighted the importance of temperature and time in mycotoxin stability in heat treatments. OTA is more stable than DON in a baking treatment. Interestingly, the DON-3-glucoside concentrations increased (>300%) under mild baking conditions. On the other hand, it was rapidly reduced under harsh conditions. The 3-ADON decreased during the heat treatment; while DOM-1 increased after the heating process. Finally, the data followed first order kinetics for analysed mycotoxins and thermal constant rates (k) were calculated. This parameter can be a useful tool for prediction of mycotoxin levels. Copyright © 2015 Elsevier Ltd. All rights reserved.
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USDA-ARS?s Scientific Manuscript database
Effect of moisture content variation on the accuracy of single kernel deoxynivalenol (DON) prediction by near-infrared (NIR) spectroscopy was investigated. Sample moisture content (MC) considerably affected accuracy of the current NIR DON calibration by underestimating or over estimating DON at high...
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Schmeitzl, Clemens; Warth, Benedikt; Fruhmann, Philipp; Michlmayr, Herbert; Malachová, Alexandra; Berthiller, Franz; Schuhmacher, Rainer; Krska, Rudolf; Adam, Gerhard
2015-01-01
Deoxynivalenol (DON) is a protein synthesis inhibitor produced by the Fusarium species, which frequently contaminates grains used for human or animal consumption. We treated a wheat suspension culture with DON or one of its acetylated derivatives, 3-acetyl-DON (3-ADON), 15-acetyl-DON (15-ADON) and 3,15-diacetyl-DON (3,15-diADON), and monitored the metabolization over a course of 96 h. Supernatant and cell extract samples were analyzed using a tailored LC-MS/MS method for the quantification of DON metabolites. We report the formation of tentatively identified DON-15-O-β-D-glucoside (D15G) and of 15-acetyl-DON-3-sulfate (15-ADON3S) as novel deoxynivalenol metabolites in wheat. Furthermore, we found that the recently identified 15-acetyl-DON-3-O-β-D-glucoside (15-ADON3G) is the major metabolite produced after 15-ADON challenge. 3-ADON treatment led to a higher intracellular content of toxic metabolites after six hours compared to all other treatments. 3-ADON was exclusively metabolized into DON before phase II reactions occurred. In contrast, we found that 15-ADON was directly converted into 15-ADON3G and 15-ADON3S in addition to metabolization into deoxynivalenol-3-O-β-D-glucoside (D3G). This study highlights significant differences in the metabolization of DON and its acetylated derivatives. PMID:26274975
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Bryła, Marcin; Ksieniewicz-Woźniak, Edyta; Waśkiewicz, Agnieszka; Szymczyk, Krystyna; Jędrzejczak, Renata
2018-01-01
The presence of mycotoxins in cereal grain is a very important food safety factor. The occurrence of “masked” mycotoxins has been intensively investigated in recent years. In this study, the occurrence of nivalenol, deoxynivalenol-3-glucoside, and deoxynivalenol in 92 samples of winter wheat from Polish cultivars was determined. The frequency of the occurrence of deoxynivalenol and nivalenol in the samples was 83% and 70%, respectively. The average content of the analytes was: for deoxynivalenol 140.2 µg/kg (10.5–1265.4 µg/kg), for nivalenol 35.0 µg/kg (5.1–372.5 µg/kg). Deoxynivalenol-3-glucoside, the formation of which is connected with the biotransformation pathway in plants, was present in 27% of tested wheat samples; its average content was 41.9 µg/kg (15.8–137.5 µg/kg). The relative content of deoxynivalenol-3-glucoside (DON-3G) compared to deoxynivalenol (DON) in positive samples was 4–37%. Despite the high frequency of occurrence of these mycotoxins, the quality of wheat from the 2016 season was good. The maximum content of DON, as defined in EU regulations (1250 µg/kg), was exceeded in only one sample. Nevertheless, the presence of a glycosidic derivative of deoxynivalenol can increase the risk to food safety, as it can be hydrolyzed by intestinal microflora. PMID:29438296
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USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON) is a toxin produced by certain species of fungi that can infest wheat, barley, and corn. The fungi cause diseases in crops worldwide and some of the secondary metabolites, such as DON, can adversely affect animal health and food safety. To rapidly monitor DON in wheat a biosenso...
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Kostelanska, Marta; Dzuman, Zbynek; Malachova, Alexandra; Capouchova, Ivana; Prokinova, Evzenie; Skerikova, Alena; Hajslova, Jana
2011-09-14
The co-occurrence of the major Fusarium mycotoxin deoxynivalenol (DON) and its conjugate deoxynivalenol-3-glucoside (DON-3-Glc) has been documented in infected wheat. This study reports on the fate of this masked DON within milling and baking technologies for the first time and compares its levels with those of the free parent toxin. The fractionation of DON-3-Glc and DON in milling fractions was similar, tested white flours contained only approximately 60% of their content in unprocessed wheat grains. No substantial changes of both target analytes occurred during the dough preparation process, i.e. kneading, fermentation, and proofing. However, when bakery improvers enzymes mixtures were employed as a dough ingredient, a distinct increase up to 145% of conjugated DON-3-Glc occurred in fermented dough. Some decrease of both DON-3-Glc and DON (10 and 13%, respectively, compared to fermented dough) took place during baking. Thermal degradation products of DON, namely norDON A, B, C, D, and DON-lactone were detected in roasted wheat samples and baked bread samples by means of UPLC-Orbitrap MS. Moreover, thermal degradation products derived from DON-3-Glc were detected and tentatively identified in heat-treated contaminated wheat and bread based on accurate mass measurement performed under the ultrahigh mass resolving power. These products, originating from DON-3-Glc through de-epoxidation and other structural changes in the seskviterpene cycle, were named norDON-3-Glc A, B, C, D, and DON-3-Glc-lactone analogically to DON degradation products. Most of these compounds were located in the crust of experimental breads.
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Generotti, Silvia; Cirlini, Martina; Malachova, Alexandra; Sulyok, Michael; Berthiller, Franz; Dall’Asta, Chiara; Suman, Michele
2015-01-01
In the scientific field, there is a progressive awareness about the potential implications of food processing on mycotoxins especially concerning thermal treatments. High temperatures may cause, in fact, transformation or degradation of these compounds. This work is aimed to study the fate of mycotoxins during bakery processing, focusing on deoxynivalenol (DON) and deoxynivalenol-3-glucoside (DON3Glc), along the chain of industrial rusk production. Starting from naturally contaminated bran, we studied how concentrations of DON and DON3Glc are influenced by modifying ingredients and operative conditions. The experiments were performed using statistical Design of Experiment (DoE) schemes to synergistically explore the relationship between mycotoxin reduction and the indicated processing transformation parameters. All samples collected during pilot plant experiments were analyzed with an LC-MS/MS multimycotoxin method. The obtained model shows a good fitting, giving back relevant information in terms of optimization of the industrial production process, in particular suggesting that time and temperature in baking and toasting steps are highly relevant for minimizing mycotoxin level in rusks. A reduction up to 30% for DON and DON3Glc content in the finished product was observed within an acceptable technological range. PMID:26213969
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Generotti, Silvia; Cirlini, Martina; Malachova, Alexandra; Sulyok, Michael; Berthiller, Franz; Dall'Asta, Chiara; Suman, Michele
2015-07-24
In the scientific field, there is a progressive awareness about the potential implications of food processing on mycotoxins especially concerning thermal treatments. High temperatures may cause, in fact, transformation or degradation of these compounds. This work is aimed to study the fate of mycotoxins during bakery processing, focusing on deoxynivalenol (DON) and deoxynivalenol-3-glucoside (DON3Glc), along the chain of industrial rusk production. Starting from naturally contaminated bran, we studied how concentrations of DON and DON3Glc are influenced by modifying ingredients and operative conditions. The experiments were performed using statistical Design of Experiment (DoE) schemes to synergistically explore the relationship between mycotoxin reduction and the indicated processing transformation parameters. All samples collected during pilot plant experiments were analyzed with an LC-MS/MS multimycotoxin method. The obtained model shows a good fitting, giving back relevant information in terms of optimization of the industrial production process, in particular suggesting that time and temperature in baking and toasting steps are highly relevant for minimizing mycotoxin level in rusks. A reduction up to 30% for DON and DON3Glc content in the finished product was observed within an acceptable technological range.
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Microbial biotransformation of DON: molecular basis for reduced toxicity
Pierron, Alix; Mimoun, Sabria; Murate, Leticia S.; Loiseau, Nicolas; Lippi, Yannick; Bracarense, Ana-Paula F. L.; Schatzmayr, Gerd; He, Jian Wei; Zhou, Ting; Moll, Wulf-Dieter; Oswald, Isabelle P.
2016-01-01
Bacteria are able to de-epoxidize or epimerize deoxynivalenol (DON), a mycotoxin, to deepoxy-deoxynivalenol (deepoxy-DON or DOM-1) or 3-epi-deoxynivalenol (3-epi-DON), respectively. Using different approaches, the intestinal toxicity of 3 molecules was compared and the molecular basis for the reduced toxicity investigated. In human intestinal epithelial cells, deepoxy-DON and 3-epi-DON were not cytotoxic, did not change the oxygen consumption or impair the barrier function. In intestinal explants, exposure for 4 hours to 10 μM DON induced intestinal lesions not seen in explants treated with deepoxy-DON and 3-epi-DON. A pan-genomic transcriptomic analysis was performed on intestinal explants. 747 probes, representing 323 genes, were differentially expressed, between DON-treated and control explants. By contrast, no differentially expressed genes were observed between control, deepoxy-DON and 3-epi-DON treated explants. Both DON and its biotransformation products were able to fit into the pockets of the A-site of the ribosome peptidyl transferase center. DON forms three hydrogen bonds with the A site and activates MAPKinases (mitogen-activated protein kinases). By contrast deepoxy-DON and 3-epi-DON only form two hydrogen bonds and do not activate MAPKinases. Our data demonstrate that bacterial de-epoxidation or epimerization of DON altered their interaction with the ribosome, leading to an absence of MAPKinase activation and a reduced toxicity. PMID:27381510
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Microbial biotransformation of DON: molecular basis for reduced toxicity
NASA Astrophysics Data System (ADS)
Pierron, Alix; Mimoun, Sabria; Murate, Leticia S.; Loiseau, Nicolas; Lippi, Yannick; Bracarense, Ana-Paula F. L.; Schatzmayr, Gerd; He, Jian Wei; Zhou, Ting; Moll, Wulf-Dieter; Oswald, Isabelle P.
2016-07-01
Bacteria are able to de-epoxidize or epimerize deoxynivalenol (DON), a mycotoxin, to deepoxy-deoxynivalenol (deepoxy-DON or DOM-1) or 3-epi-deoxynivalenol (3-epi-DON), respectively. Using different approaches, the intestinal toxicity of 3 molecules was compared and the molecular basis for the reduced toxicity investigated. In human intestinal epithelial cells, deepoxy-DON and 3-epi-DON were not cytotoxic, did not change the oxygen consumption or impair the barrier function. In intestinal explants, exposure for 4 hours to 10 μM DON induced intestinal lesions not seen in explants treated with deepoxy-DON and 3-epi-DON. A pan-genomic transcriptomic analysis was performed on intestinal explants. 747 probes, representing 323 genes, were differentially expressed, between DON-treated and control explants. By contrast, no differentially expressed genes were observed between control, deepoxy-DON and 3-epi-DON treated explants. Both DON and its biotransformation products were able to fit into the pockets of the A-site of the ribosome peptidyl transferase center. DON forms three hydrogen bonds with the A site and activates MAPKinases (mitogen-activated protein kinases). By contrast deepoxy-DON and 3-epi-DON only form two hydrogen bonds and do not activate MAPKinases. Our data demonstrate that bacterial de-epoxidation or epimerization of DON altered their interaction with the ribosome, leading to an absence of MAPKinase activation and a reduced toxicity.
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A new perspective on deoxynivalenol and growth suppression
USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON) is a trichothecene produced by Fusarium species. It is found in cereal grains, feeds and foods and exerts various toxic effects in farm and laboratory animals. These include vomiting, loss of appetite, and growth suppression. Surveys have shown that DON intake by consumers, i...
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Rapid Detection of Nivalenol and Deoxynivalenol in Wheat Using Surface Plasmon Resonance Immunoassay
USDA-ARS?s Scientific Manuscript database
Surface plasmon resonance immunoassay using a monoclonal antibody was developed to measure nivalenol (NIV) and deoxynivalenol (DON) contamination in wheat. A DON-immobilized sensor chip having high sensitivity and stability was prepared, and an SPR detection procedure was developed. The competitiv...
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Amarasinghe, Chami C; Simsek, Senay; Brûlé-Babel, Anita; Fernando, W G Dilantha
2016-07-01
Contamination of wheat grains with Fusarium mycotoxins and their modified forms is an important issue in wheat industry. The objective of this study was to analyse the deoxynivalenol (DON) and deoxynivalenol-3-glucosides (D3G) content in Canadian spring wheat cultivars grown in two locations, inoculated with a mixture of 3-acetyldeoxynivalenol (3-ADON)-producing Fusarium graminearum strains and a mixture of 15-acetlyldeoxynivalenol (15-ADON)-producing F. graminearum strains. According to the analysis of variance, significant differences were observed among the cultivars for Fusarium head blight (FHB) disease index, Fusarium-damaged kernel percentage (%FDK), DON content and D3G content. When the effect of chemotype was considered, significant differences were observed for FHB disease index, FDK percentage and DON content. The D3G content and D3G/DON ratio were not significantly different between the chemotypes, except for D3G content at the Winnipeg location. The Pearson correlation coefficient between DON and D3G was 0.84 and 0.77 at Winnipeg and Carman respectively. The highest D3G/DON ratio was observed in cultivars Carberry (44%) in Carman and CDC Kernen (63.8%) in Winnipeg. The susceptible cultivars showed lower D3G/DON ratio compared with the cultivars rated as moderately resistant and intermediate. The current study indicated that Canadian spring cultivars produce D3G upon Fusarium infection.
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Using enzymes and microorganisms to modify the mycotoxin deoxynivalenol
USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON) is a trichothecene mycotoxin produced by the fungus Fusarium graminearum that contaminates staple crops such as wheat, barley, and maize when they are infected with this fungus. New strategies are needed to mitigate DON. We screened for microbes that could grow in the presence o...
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USDA-ARS?s Scientific Manuscript database
The trichothecene mycotoxin deoxynivalenol (DON) is a common contaminant of wheat, barley, and maize. New strategies are needed to reduce or eliminate DON in feed and food products. Microorganisms from plant and soil samples collected in Blacksburg, VA, USA, were screened by incubation in a mineral ...
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Wu, Qinghua; Kuča, Kamil; Humpf, Hans-Ulrich; Klímová, Blanka; Cramer, Benedikt
2017-02-01
Deoxynivalenol (DON), the most commonly occurring trichothecene in nature, may affect animal and human health through causing diarrhea, vomiting, gastrointestinal inflammation, and immunomodulation. DON-3-glucoside (DON-3G) as a major plant metabolite of the mycotoxin is another "emerging" food safety issue in recent years. Humans may experience potential health risks by consuming DON-contaminated food products. Thus, it is crucial for human and animal health to study also the degradation of DON and DON-3G during thermal food processing. Baking, boiling, steaming, frying, and extrusion cooking are commonly used during thermal food processing and have promising effects on the reduction of mycotoxins in food. For DON, however, the observed effects of these methods, as reported in numerous studies, are ambiguous and do not present a clear picture with regard to reduction or transformation. This review summarized the influence of thermal processing on the stability of DON and the formation of degradation/conversion products. Besides this, also a release of DON and DON-3G from food matrix as well as the release of DON from DON-3G during processing is discussed. In addition, some conflicting findings as reported from the studies on thermal processing as well as cause-effect relationships of the different thermal procedures are explored. Finally, the potential toxic profiles of DON degradation products are discussed as well when data are available.
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USDA-ARS?s Scientific Manuscript database
Rapid and routine detection of deoxynivalenol (DON) in cereals-based food and feed has long been a strong desire of regulators and manufacturers. Traditional chemical methods and antibody based biosensors and immunoassays have been developed as viable tools to identify and measure DON. However, thes...
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USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON)is a mycotoxin produced by certain fungi that infest cereal grains worldwide. A hybridoma cell line producing a monoclonal antibody (Mab) recognizing DON was used as the starting point in the development of a recombinant single chain variable fragment (scFv) antibody. The scFv wa...
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USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON) levels in harvested grain samples are used to evaluate the Fusarium head blight (FHB) resistance of wheat cultivars and breeding lines. Fourier transform near-infrared (FT-NIR) calibrations were developed to estimate the DON and moisture content (MC) of bulk wheat grain samples ...
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Deoxynivalenol Biomarkers in the Urine of UK Vegetarians.
Wells, Liz; Hardie, Laura; Williams, Courtney; White, Kay; Liu, Yunru; De Santis, Barbara; Debegnach, Francesca; Moretti, Georgio; Greetham, Stephanie; Brera, Carlo; Papageorgiou, Maria; Thatcher, Natalie J; Rigby, Alan; Atkin, Stephen L; Sathyapalan, Thozhukat
2017-06-22
Deoxynivalenol (DON) is produced by Fusarium graminearum and is one of the most commonly occurring trichothecenes. Vegetarians are alleged to be a high-risk group for DON exposure due to high intakes of cereals susceptible to the growth of the mycotoxin. This study provides the levels of DON and de-epoxi Deoxynivalenol (DOM-1) in urine analysed by liquid chromatography-mass spectrometry (LC-MS) in UK vegetarians. Over two consecutive days, morning urine samples were collected from 32 vegetarians and 31 UK adult volunteers, and associated food consumption 24 h prior to the sample was recorded. Statistically significant differences between the weight of the UK adults and vegetarians ( t = 3.15. df = 61, p ≤ 0.005 two-tailed) were observed. The mean levels of DON in urine for adults on day 1 was 3.05 ng free DON/mg creatinine, and on day 2 was 2.98 ng free DON/mg creatinine. Even though high mean levels were observed, most adults were within the tolerable daily intake. However, for vegetarians, the mean level of urinary DON on day 1 was 6.69 ng free DON/mg creatinine, and on day 2 was 3.42 ng free DON/mg creatinine. These levels equate to up to 32% of vegetarians exceeding recommended tolerable daily intakes (TDI) of exposure (1 µg/kg b.w./day).
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Determination of Deoxynivalenol in the Urine of Pregnant Women in the UK
Wells, Liz; Hardie, Laura; Williams, Courtney; White, Kay; Liu, Yunru; De Santis, Barbara; Debegnach, Francesca; Moretti, Georgio; Greetham, Stephanie; Brera, Carlo; Rigby, Alan; Atkin, Stephen; Sathyapalan, Thozhukat
2016-01-01
Deoxynivalenol (DON) is one of the most commonly occurring trichothecenes, produced mainly by Fusarium graminearum. Little is known about the effect of DON exposure or the levels of DON exposure that occur during pregnancy. The project aimed to provide data on levels of total DON and de-epoxi Deoxynivalenol (DOM-1) in pregnant human urine samples analysed by liquid chromatography-mass spectrometry (LC-MS). Morning urine samples were collected over two consecutive days from 42 volunteers and associated food consumption was recorded for the 24 h prior to the sample. Spearman’s rho non-parametric test for correlation was used to assess the data. Levels of DON did not differ significantly between day 1 (mean 29.7 ng/mL urine or 40.1 ng DON/mg creatinine) and day 2 (mean 28.7 ng/mL urine or 38.8 ng DON/mg creatinine ng/mL/day) urine samples. The only significant positive correlation was found between total ng DON/mg creatinine and parity (rho = 0.307, n = 42, p < 0.005 two-tailed) and total ng DON/mg creatinine with baked goods on day 1 (rho = 0.532, n = 42, p < 0.0005 two-tailed). This study provides data on the DON levels in pregnancy in this suburban population and reassurance that those levels are within acceptable limits. PMID:27792137
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USDA-ARS?s Scientific Manuscript database
A sample preparation method was developed for the screening of deoxynivalenol (DON) in wheat and wheat dust. Extraction was carried out with water and was successful due to the polar character of DON. For detection, an enzyme-linked immunosorbent assay (ELISA) was compared to the sensor-based techni...
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Schwartz-Zimmermann, Heidi E.; Fruhmann, Philipp; Dänicke, Sven; Wiesenberger, Gerlinde; Caha, Sylvia; Weber, Julia; Berthiller, Franz
2015-01-01
Recently, deoxynivalenol-3-sulfate (DON-3-sulfate) was proposed as a major DON metabolite in poultry. In the present work, the first LC-MS/MS based method for determination of DON-3-sulfate, deepoxy-DON-3-sulfate (DOM-3-sulfate), DON, DOM, DON sulfonates 1, 2, 3, and DOM sulfonate 2 in excreta samples of chickens and turkeys was developed and validated. To this end, DOM-3-sulfate was chemically synthesized and characterized by NMR and LC-HR-MS/MS measurements. Application of the method to excreta and chyme samples of four feeding trials with turkeys, chickens, pullets, and roosters confirmed DON-3-sulfate as the major DON metabolite in all poultry species studied. Analogously to DON-3-sulfate, DOM-3-sulfate was formed after oral administration of DOM both in turkeys and in chickens. In addition, pullets and roosters metabolized DON into DOM-3-sulfate. In vitro transcription/translation assays revealed DOM-3-sulfate to be 2000 times less toxic on the ribosome than DON. Biological recoveries of DON and DOM orally administered to broiler chickens, turkeys, and pullets were 74%–106% (chickens), 51%–72% (roosters), and 131%–151% (pullets). In pullets, DON-3-sulfate concentrations increased from jejunum chyme samples to excreta samples by a factor of 60. This result, put into context with earlier studies, indicates fast and efficient absorption of DON between crop and jejunum, conversion to DON-3-sulfate in intestinal mucosa, liver, and possibly kidney, and rapid elimination into excreta via bile and urine. PMID:26569307
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Lactobacillus plantarum culture supernatants improve intestinal tissue exposed to deoxynivalenol.
Maidana, L G; Gerez, J; Pinho, F; Garcia, S; Bracarense, A P F L
2017-10-02
In the present study, histological, morphometrical and ultrastructural analysis were performed to investigate intestinal mucosa changes in piglets exposed to deoxynivalenol alone or associated with two strains of Lactobacillus plantarum and the respective culture supernatants. Jejunal explants were incubated for 4h in culture medium with a) only culture medium (DMEM, control group), b) deoxynivalenol (DON, 10μM), c) heat-inactivated Lactobacillus plantarum strain1 - LP1 (1.1×10 8 CFU/ml) plus DON, d) heat-inactivated Lactobacillus plantarum strain2-LP2 (2.0×10 9 CFU/ml) plus DON, e) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1) plus DON, and f) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1) plus DON. Explants exposed to DON and DON plus LP1 and LP2 showed a significant increase in histological changes (mainly villi atrophy and apical necrosis) and a significant decrease in villi height when compared to unexposed explants. However, explants treated with CS1+DON and CS2+DON remained similar to the control group both in histological and morphometrical aspects. DON also induced a significant decrease in goblet cell density compared to control whereas CS1+DON treatment induced an increase in the number of goblet cells in comparison to DON explants. In addition, ultrastructural assessment showed control, CS1+DON and CS2+DON explants with well delineated finger shape villi, meanwhile DON-treated, LP1+DON and LP2+DON explants showed a severe villi atrophy with leukocytes exudation on the intestinal surface. Taken together, our results indicate that the culture supernatant treatment reduced the toxic effects induced by DON on intestinal tissue and may contribute as an alternative strategy to reduce mycotoxin toxicity. Copyright © 2017 Elsevier GmbH. All rights reserved.
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Deoxynivalenol Exposure in Norway, Risk Assessments for Different Human Age Groups.
Sundheim, Leif; Lillegaard, Inger Therese; Fæste, Christiane Kruse; Brantsæter, Anne-Lise; Brodal, Guro; Eriksen, Gunnar Sundstøl
2017-02-04
Deoxynivalenol (DON) is the most common mycotoxin in Norwegian cereals, and DON is detected in most samples of crude cereal grain and cereal food commodities such as flour, bran, and oat flakes. The Norwegian Scientific Committee for Food Safety assessed the risk for adverse effects of deoxynivalenol (DON) in different age groups of the domestic population. This review presents the main results from the risk assessment, supplemented with some recently published data. Impairment of the immune system together with reduced feed intake and weight gain are the critical effects of DON in experimental animals on which the current tolerable daily intake was established. Based on food consumption and occurrence data, the mean exposure to DON in years with low and high levels of DON in the flour, respectively, were in the range of or up to two times the Tolerable Daily Intake (TDI) in 1-year-old infants and 2-year-old children. In years with high mean DON concentration, the high (95th-percentile) exposure exceeded the TDI by up to 3.5 times in 1-, 2- , 4-, and 9-year-old children. The assessment concluded that exceeding the TDI in infants and children is of concern. The estimated dietary DON intakes in adolescent and adult populations are in the range of the TDI or below, and are not a health concern. Acute human exposure to DON is not of concern in any age group.
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Deoxynivalenol Biomarkers in the Urine of UK Vegetarians
Wells, Liz; Hardie, Laura; Williams, Courtney; White, Kay; Liu, Yunru; De Santis, Barbara; Debegnach, Francesca; Moretti, Georgio; Greetham, Stephanie; Brera, Carlo; Papageorgiou, Maria; Thatcher, Natalie J.; Rigby, Alan; Atkin, Stephen L.; Sathyapalan, Thozhukat
2017-01-01
Deoxynivalenol (DON) is produced by Fusarium graminearum and is one of the most commonly occurring trichothecenes. Vegetarians are alleged to be a high-risk group for DON exposure due to high intakes of cereals susceptible to the growth of the mycotoxin. This study provides the levels of DON and de-epoxi Deoxynivalenol (DOM-1) in urine analysed by liquid chromatography-mass spectrometry (LC-MS) in UK vegetarians. Over two consecutive days, morning urine samples were collected from 32 vegetarians and 31 UK adult volunteers, and associated food consumption 24 h prior to the sample was recorded. Statistically significant differences between the weight of the UK adults and vegetarians (t = 3.15. df = 61, p ≤ 0.005 two-tailed) were observed. The mean levels of DON in urine for adults on day 1 was 3.05 ng free DON/mg creatinine, and on day 2 was 2.98 ng free DON/mg creatinine. Even though high mean levels were observed, most adults were within the tolerable daily intake. However, for vegetarians, the mean level of urinary DON on day 1 was 6.69 ng free DON/mg creatinine, and on day 2 was 3.42 ng free DON/mg creatinine. These levels equate to up to 32% of vegetarians exceeding recommended tolerable daily intakes (TDI) of exposure (1 µg/kg b.w./day). PMID:28640201
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Cirlini, Martina; Generotti, Silvia; Dall'Erta, Andrea; Lancioni, Pietro; Ferrazzano, Gianluca; Massi, Andrea; Galaverna, Gianni; Dall'Asta, Chiara
2013-12-24
Deoxynivalenol (DON) is the most prevalent trichothecene in Europe and its occurrence is associated with infections of Fusarium graminearum and F. culmorum, causal agents of Fusarium head blight (FHB) on wheat. Resistance to FHB is a complex character and high variability occurs in the relationship between DON content and FHB incidence. DON conjugation to glucose (DON-3-glucoside, D3G) is the primary plant mechanism for resistance towards DON accumulation. Although this mechanism has been already described in bread wheat and barley, no data are reported so far about durum wheat, a key cereal in the pasta production chain. To address this issue, the ability of durum wheat to detoxify and convert deoxynivalenol into D3G was studied under greenhouse controlled conditions. Four durum wheat varieties (Svevo, Claudio, Kofa and Neodur) were assessed for DON-D3G conversion; Sumai 3, a bread wheat variety carrying a major QTL for FHB resistance (QFhs.ndsu-3B), was used as a positive control. Data reported hereby clearly demonstrate the ability of durum wheat to convert deoxynivalenol into its conjugated form, D3G.
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USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON) is a mycotoxin which isproduced by plant pathogens such as Fusarium species. The formation of the "masked" mycotoxin deoxinyvalenol-3-glucoside (D3G) results from a defense mechanism the plant uses for detoxification. These two mycotoxins are important from the food safety poi...
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Deoxynivalenol Exposure in Norway, Risk Assessments for Different Human Age Groups
Sundheim, Leif; Lillegaard, Inger Therese; Fæste, Christiane Kruse; Brantsæter, Anne-Lise; Brodal, Guro; Eriksen, Gunnar Sundstøl
2017-01-01
Deoxynivalenol (DON) is the most common mycotoxin in Norwegian cereals, and DON is detected in most samples of crude cereal grain and cereal food commodities such as flour, bran, and oat flakes. The Norwegian Scientific Committee for Food Safety assessed the risk for adverse effects of deoxynivalenol (DON) in different age groups of the domestic population. This review presents the main results from the risk assessment, supplemented with some recently published data. Impairment of the immune system together with reduced feed intake and weight gain are the critical effects of DON in experimental animals on which the current tolerable daily intake was established. Based on food consumption and occurrence data, the mean exposure to DON in years with low and high levels of DON in the flour, respectively, were in the range of or up to two times the Tolerable Daily Intake (TDI) in 1-year-old infants and 2-year-old children. In years with high mean DON concentration, the high (95th-percentile) exposure exceeded the TDI by up to 3.5 times in 1-, 2- , 4-, and 9-year-old children. The assessment concluded that exceeding the TDI in infants and children is of concern. The estimated dietary DON intakes in adolescent and adult populations are in the range of the TDI or below, and are not a health concern. Acute human exposure to DON is not of concern in any age group. PMID:28165414
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The fate and tissue disposition of deoxynivalenol in broiler chickens
PRALATNET, Sasithorn; POAPOLATHEP, Saranya; IMSILP, Kanjana; TANHAN, Phanwimol; ISARIYODOM, Supaporn; KUMAGAI, Susumu; POAPOLATHEP, Amnart
2015-01-01
To evaluate the fate of deoxynivalenol (DON) in broilers, DON was administered either intravenously or orally to broilers at a dose of 1 mg/kg BW. Concentrations of DON in plasma were measurable up to 4 hr and 2 hr after intravenous and oral administration, respectively. Following intravenous administration, the values for the elimination half-life, the volume of distribution and the clearance were 1.25 ± 0.25 hr, 7.55 ± 2.03 l/kg and 4.16 ± 0.42 l/hr/kg, respectively. The oral bioavailability was 15.46 ± 4.02%. DON was detectable in all tissues examined after oral administration. These results suggest that DON is able to penetrate into the various tissues in broilers, though poorly absorbed from their gastrointestinal tract. PMID:25843039
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Tian, Ye; Tan, Yanglan; Liu, Na; Yan, Zheng; Liao, Yucai; Chen, Jie; de Saeger, Sarah; Yang, Hua; Zhang, Qiaoyan; Wu, Aibo
2016-11-15
Deoxynivalenol (DON) is a mycotoxin mainly produced by the Fusarium graminearum complex, which are important phytopathogens that can infect crops and lead to a serious disease called Fusarium head blight (FHB). As the most common B type trichothecene mycotoxin, DON has toxic effects on animals and humans, which poses a risk to food security. Thus, efforts have been devoted to control DON contamination in different ways. Management of DON production by Trichoderma strains as a biological control-based strategy has drawn great attention recently. In our study, eight selected Trichoderma strains were evaluated for their antagonistic activities on F. graminearum by dual culture on potato dextrose agar (PDA) medium. As potential antagonists, Trichoderma strains showed prominent inhibitory effects on mycelial growth and mycotoxin production of F. graminearum . In addition, the modified mycotoxin deoxynivalenol-3-glucoside (D3G), which was once regarded as a detoxification product of DON in plant defense, was detected when Trichoderma were confronted with F. graminearum . The occurrence of D3G in F. graminearum and Trichoderma interaction was reported for the first time, and these findings provide evidence that Trichoderma strains possess a self-protection mechanism as plants to detoxify DON into D3G when competing with F. graminearum .
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Tian, Ye; Tan, Yanglan; Liu, Na; Yan, Zheng; Liao, Yucai; Chen, Jie; de Saeger, Sarah; Yang, Hua; Zhang, Qiaoyan; Wu, Aibo
2016-01-01
Deoxynivalenol (DON) is a mycotoxin mainly produced by the Fusarium graminearum complex, which are important phytopathogens that can infect crops and lead to a serious disease called Fusarium head blight (FHB). As the most common B type trichothecene mycotoxin, DON has toxic effects on animals and humans, which poses a risk to food security. Thus, efforts have been devoted to control DON contamination in different ways. Management of DON production by Trichoderma strains as a biological control-based strategy has drawn great attention recently. In our study, eight selected Trichoderma strains were evaluated for their antagonistic activities on F. graminearum by dual culture on potato dextrose agar (PDA) medium. As potential antagonists, Trichoderma strains showed prominent inhibitory effects on mycelial growth and mycotoxin production of F. graminearum. In addition, the modified mycotoxin deoxynivalenol-3-glucoside (D3G), which was once regarded as a detoxification product of DON in plant defense, was detected when Trichoderma were confronted with F. graminearum. The occurrence of D3G in F. graminearum and Trichoderma interaction was reported for the first time, and these findings provide evidence that Trichoderma strains possess a self-protection mechanism as plants to detoxify DON into D3G when competing with F. graminearum. PMID:27854265
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Ji, Fang; Wu, Jirong; Zhao, Hongyan; Xu, Jianhong; Shi, Jianrong
2015-03-05
A total of 122 wheat varieties obtained from the Nordic Genetic Resource Center were infected artificially with an aggressive Fusariumasiaticum strain in a field experiment. We calculated the severity of Fusarium head blight (FHB) and determined the deoxynivalenol (DON) content of wheat grain, straw and glumes. We found DON contamination levels to be highest in the glumes, intermediate in the straw, and lowest in the grain in most samples. The DON contamination levels did not increase consistently with increased FHB incidence. The DON levels in the wheat varieties with high FHB resistance were not necessarily low, and those in the wheat varieties with high FHB sensitivity were not necessarily high. We selected 50 wheat genotypes with reduced DON content for future research. This study will be helpful in breeding new wheat varieties with low levels of DON accumulation.
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Li, Yuqin; Wang, Hao; Jia, Baoxiu; Liu, Caihong; Liu, Ke; Qi, Yongxiu; Hu, Zhide
2013-01-01
The mechanism of interaction between deoxynivalenol (DON) and human serum albumin (HSA) was studied using spectroscopic methods including fluorescence spectra, UV-VIS, Fourier transform infrared (FT-IR) and circular dichroism (CD). The quenching mechanism was investigated in terms of the association constants, number of binding sites and basic thermodynamic parameters. The distance between the HSA donor and the acceptor DON was 2.80 nm as derived from fluorescence resonance energy transfer. The secondary structure compositions of free HSA and its DON complexes were estimated by the FT-IR spectra. Alteration of the secondary protein structure in the presence of DON was confirmed by UV-VIS and CD spectroscopy. Molecular modelling revealed that a DON-protein complex was stabilised by hydrophobic forces and hydrogen bonding. It was potentially useful for elucidating the toxigenicity of DON when combined with biomolecular function effect, transmembrane transport, toxicological testing and the other experiments.
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Functional Agents to Biologically Control Deoxynivalenol Contamination in Cereal Grains
Tian, Ye; Tan, Yanglan; Liu, Na; Liao, Yucai; Sun, Changpo; Wang, Shuangxia; Wu, Aibo
2016-01-01
Mycotoxins, as microbial secondary metabolites, frequently contaminate cereal grains and pose a serious threat to human and animal health around the globe. Deoxynivalenol (DON), a commonly detected Fusarium mycotoxin, has drawn utmost attention due to high exposure levels and contamination frequency in the food chain. Biological control is emerging as a promising technology for the management of DON contamination. Functional biological control agents (BCAs), which include antagonistic microbes, natural fungicides derived from plants and detoxification enzymes, can be used to control DON contamination at different stages of grain production. In this review, studies regarding different biological agents for DON control in recent years are summarized for the first time. Furthermore, this article highlights the significance of BCAs for controlling DON contamination, as well as the need for more practical and efficient BCAs concerning food safety. PMID:27064760
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Genotoxic effects of deoxynivalenol in broiler chickens fed low-protein feeds.
Awad, W A; Ghareeb, K; Dadak, A; Gille, L; Staniek, K; Hess, M; Böhm, J
2012-03-01
Deoxynivalenol (DON) is one of the most abundant and important trichothecenes in food and feed, and it is a significant contaminant due to its frequent occurrence at toxicologically relevant concentrations worldwide. Deoxynivalenol has negative influences on the health and performance of chicks. However, there is little information available regarding the effect of DON on DNA fragmentation in blood lymphocytes. In addition, the effects of Mycofix select (Biomin GmbH, Herzogenburg, Austria) supplementation to DON-contaminated broiler diets on lymphocyte DNA have not yet been demonstrated. Therefore, the aim of the present study was to establish the effect of DON on lipid peroxidation and lymphocyte DNA fragmentation in broilers and to evaluate the potential of Mycofix select in the prevention of toxin-mediated changes. Thirty-two 1-d-old (Ross 308 male) broiler chicks were randomly divided into 4 groups. The control group was fed a noncontaminated diet, and a second group was fed the same diet but supplemented with Mycofix select (0.25%). A third group of broilers was fed a diet artificially contaminated with 10 mg of feed-grade DON/kg of diet, and a fourth group was fed a DON-contaminated diet supplemented with Mycofix select. At the end of the feeding trial, blood was collected and the degree of lymphocyte DNA damage was measured in the plasma by comet assay. Deoxynivalenol increased (P = 0.016) the amount of DNA damage in chicken lymphocytes by 46.8%. Mycofix select protected lymphocyte DNA from the DON effects. To our knowledge, these are the first data on genotoxic effects of a moderate dose of DON on chicken lymphocytes. However, the thiobarbituric acid reactive substances level in liver and liver enzyme activity did not differ among the groups. In conclusion, the present study demonstrated that the diets contaminated with the mycotoxin DON at moderate levels in combination with low-protein feed are able to induce lymphocyte DNA damage in chickens. Supplementation with Mycofix select protected lymphocyte DNA and it was beneficial for maintaining the lymphocyte DNA integrity.
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Dietary l-Arginine Supplementation Protects Weanling Pigs from Deoxynivalenol-Induced Toxicity
Wu, Li; Liao, Peng; He, Liuqin; Feng, Zemeng; Ren, Wenkai; Yin, Jie; Duan, Jielin; Li, Tiejun; Yin, Yulong
2015-01-01
This study was conducted to determine the positive effects of dietary supplementation with l-arginine (Arg) on piglets fed a deoxynivalenol (DON)-contaminated diet. A total of eighteen, 28-day-old healthy weanling pigs were randomly assigned into one of three groups: uncontaminated basal diet (control group), 6 mg/kg DON-contaminated diet (DON group) and 6 mg/kg DON + 1% l-arginine (DON + ARG group). After 21 days of Arg supplementation, piglets in the DON and DON + ARG groups were challenged by feeding 6 mg/kg DON-contaminated diet for seven days. The results showed that DON resulted in damage to piglets. However, clinical parameters, including jejunal morphology, amino acid concentrations in the serum, jejunum and ileum, were improved by Arg (p < 0.05). Furthermore, the mRNA levels for sodium-glucose transporter-1 (SGLT-1), glucose transporter type-2 (GLUT-2) and y+l-type amino acid transporter-1 (y+LAT-1) were downregulated in the DON group, but the values were increased in the DON + ARG group (p < 0.05). Collectively, these results indicate that dietary supplementation with Arg exerts a protective role in pigs fed DON-contaminated diets. PMID:25884909
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[Pollution investigation of deoxynivalenol in wheat flour of China in 2013].
Lu, Jingjing; Yang, Dajin
2015-07-01
To study the deoxynivalenol (DON) contamination status in wheat flour of China in 2013. Stereotypes packaged or bulk wheat flour samples sold in 28 provinces were collected in a random sampling way. The concentration of DON in each flour wheat sample was measured by high performance liquid chromatography. The results were statistically analyzed and evaluated. A total of 5678 wheat flour was detected. The detection rate of DON was 58.74%. The excessive rate of the standard of DON was 4.60%. The average content of DON was 317 µg/kg. The content range of DON was 0-56.1 mg/kg. DON pollution is relatively common in wheat flour of China in 2013, but the excessive rate is not high. The degree of pollution in each area is different. The excessive rate of DON, which was associated with the local temperature and humidity conditions, in wheat flour sold in east, southwest and northwest area is relatively high. Pollution level of DON in wheat flour in 2013 is consistent with those in 2010 and 2011, but lower than the monitoring results in 2012.
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Deoxynivalenol and its acetyl derivatives in bread and biscuits in Shandong province of China.
Jiang, Dafeng; Chen, Jindong; Li, Fenghua; Li, Wei; Yu, Lianlong; Zheng, Fengjia; Wang, Xiaolin
2018-03-01
In the present study, 100 commercial breads and biscuits were investigated for the occurrence of deoxynivalenol (DON) and its acetylated derivatives 3-acetyldeoxynivalenol (3-Ac-DON) and 15-acetyldeoxynivalenol (15-Ac-DON). The target mycotoxins were determined by isotope dilution ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). DON was determined in 95% of the test samples with a mean value of 153.3 µg/kg. Compared with DON, 3-Ac-DON and 15-Ac-DON were far less frequently detected, with mean values of 1.14 and 0.37 µg/kg, respectively. The estimated daily intakes of the sum of DON and its derivatives in breads and biscuits were 0.0059 and 0.0313 µg/kg bw/day, respectively, which was within the group provisional tolerable daily intake of 1.0 µg/kg bw/day set by the Joint FAO/WHO Expert Committee on Food Additives. In the future, systematic monitoring programmes of DON and its derivatives in relevant foodstuffs are still necessary for food safety and human health.
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Novak, Barbara; Vatzia, Eleni; Springler, Alexandra; Pierron, Alix; Reisinger, Nicole; Hessenberger, Sabine; Mayer, Elisabeth
2018-01-01
Deoxynivalenol (DON) is one of the most prevalent mycotoxins, contaminating cereals and cereal-derived products. Its derivative deepoxy-deoxynivalenol (DOM-1) is produced by certain bacteria, which either occur naturally or are supplemented in feed additive. DON-induced impairments in protein synthesis are particularly problematic for highly proliferating immune cells. This study provides the first comparison of the effects of DON and DOM-1 on the concanavalin A-induced proliferation of porcine, chicken, and bovine peripheral blood mononuclear cells (PBMCs). Therefore, isolated PBMCs were treated with DON (0.01–3.37 µM) and DOM-1 (1.39–357 µM) separately, and proliferation was measured using a bromodeoxyuridine (BrdU) assay. Although pigs are considered highly sensitive to DON, the present study revealed a substantially higher sensitivity of bovine (IC50 = 0.314 µM) PBMCs compared to chicken (IC50 = 0.691 µM) and porcine (IC50 = 0.693 µM) PBMCs. Analyses on the proliferation of bovine T-cell subsets showed that all major subsets, namely, CD4+, CD8β+, and γδ T cells, were affected to a similar extent. In contrast, DOM-1 did not affect bovine PBMCs, but reduced the proliferation of chicken and porcine PBMCs at the highest tested concentration (357 µM). Results confirm the necessity of feed additives containing DON-to-DOM-1-transforming bacteria and highlights species-specific differences in the DON sensitivity of immune cells. PMID:29641442
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USDA-ARS?s Scientific Manuscript database
The fungus Fusarium graminearum produces the toxic compound deoxynivalenol (DON) that contaminates wheat, barley, and maize. New strategies are needed to mitigate DON in the United States. Microbes were isolated from different soil types, and cultured in a mineral salt media using 100 ppm DON as the...
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Factors Influencing Deoxynivalenol Accumulation in Small Grain Cereals
Wegulo, Stephen N.
2012-01-01
Deoxynivalenol (DON) is a mycotoxin produced by the plant pathogenic fungi Fusarium graminearum and F. culmorum. These and other closely related fungi cause a disease known as Fusarium head blight (FHB) in small grain cereals. Other mycotoxins produced by FHB-causing fungi include nivalenol, T-2 toxin, and zearalenone. Ingestion of mycotoxin-contaminated food and feed can lead to toxicosis in humans and animals, respectively. DON is the predominant and most economically important of these mycotoxins in the majority of small grain-producing regions of the world. This review examines the factors that influence DON accumulation in small grain cereals from an agricultural perspective. The occurrence and economic importance of FHB and DON in small grain cereals, epidemiological factors and cereal production practices that favor FHB development and DON accumulation in grain under field conditions, and regulatory/advisory standards for DON in food and feed are discussed. This information can be used to develop strategies that reduce DON accumulation in grain before harvest and to mitigate the human and animal health risks associated with DON contamination of food and feed. PMID:23202310
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Sex Is a Determinant for Deoxynivalenol Metabolism and Elimination in the Mouse
Pestka, James J.; Clark, Erica S.; Schwartz-Zimmermann, Heidi E.
2017-01-01
Based on prior observations that deoxynivalenol (DON) toxicity is sex-dependent, we compared metabolism and clearance of this toxin in male and female mice. Following intraperitoneal challenge with 1 mg/kg bw DON, the dose used in the aforementioned toxicity study, ELISA and LC–MS/MS analyses revealed that by 24 h, most DON and DON metabolites were excreted via urine (49–86%) as compared to feces (1.2–8.3%). Females excreted DON and its principal metabolites (DON-3-, DON-8,15 hemiketal-8-, and iso-DON-8-glucuronides) in urine more rapidly than males. Metabolite concentrations were typically 2 to 4 times higher in the livers and kidneys of males than females from 1 to 4 h after dosing. Trace levels of DON-3-sulfate and DON-15-sulfate were found in urine, liver and kidneys from females but not males. Fecal excretion of DON and DON sulfonates was approximately 2-fold greater in males than females. Finally, decreased DON clearance rates in males could not be explained by glucuronidation activities in liver and kidney microsomes. To summarize, increased sensitivity of male mice to DON’s toxic effects as compared to females corresponds to decreased ability to clear the toxin via urine but did not appear to result from differences in toxin metabolism. PMID:28777306
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Pietsch, Constanze; Schulz, Carsten; Rovira, Pere; Kloas, Werner; Burkhardt-Holm, Patricia
2014-01-01
Deoxynivalenol (DON) frequently contaminates animal feed, including fish feed used in aquaculture. This study intends to further investigate the effects of DON on carp (Cyprinus carpio L.) at concentrations representative for commercial fish feeds. Experimental feeding with 352, 619 or 953 μg DON kg−1 feed resulted in unaltered growth performance of fish during six weeks of experimentation, but increased lipid peroxidation was observed in liver, head kidney and spleen after feeding of fish with the highest DON concentration. These effects of DON were mostly reversible by two weeks of feeding the uncontaminated control diet. Histopathological scoring revealed increased liver damage in DON-treated fish, which persisted even after the recovery phase. At the highest DON concentration, significantly more fat, and consequently, increased energy content, was found in whole fish body homogenates. This suggests that DON affects nutrient metabolism in carp. Changes of lactate dehydrogenase (LDH) activity in kidneys and muscle and high lactate levels in serum indicate an effect of DON on anaerobic metabolism. Serum albumin was reduced by feeding the medium and a high dosage of DON, probably due to the ribotoxic action of DON. Thus, the present study provides evidence of the effects of DON on liver function and metabolism. PMID:24566729
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Pan, Xiao; Center for Integrative Toxicology, Michigan State University, East Lansing, MI 48824; Whitten, Douglas A.
Deoxynivalenol (DON), a trichothecene mycotoxin produced by Fusarium that commonly contaminates food, is capable of activating mononuclear phagocytes of the innate immune system via a process termed the ribotoxic stress response (RSR). To encapture global signaling events mediating RSR, we quantified the early temporal (≤ 30 min) phosphoproteome changes that occurred in RAW 264.7 murine macrophage during exposure to a toxicologically relevant concentration of DON (250 ng/mL). Large-scale phosphoproteomic analysis employing stable isotope labeling of amino acids in cell culture (SILAC) in conjunction with titanium dioxide chromatography revealed that DON significantly upregulated or downregulated phosphorylation of 188 proteins at bothmore » known and yet-to-be functionally characterized phosphosites. DON-induced RSR is extremely complex and goes far beyond its prior known capacity to inhibit translation and activate MAPKs. Transcriptional regulation was the main target during early DON-induced RSR, covering over 20% of the altered phosphoproteins as indicated by Gene Ontology annotation and including transcription factors/cofactors and epigenetic modulators. Other biological processes impacted included cell cycle, RNA processing, translation, ribosome biogenesis, monocyte differentiation and cytoskeleton organization. Some of these processes could be mediated by signaling networks involving MAPK-, NFκB-, AKT- and AMPK-linked pathways. Fuzzy c-means clustering revealed that DON-regulated phosphosites could be discretely classified with regard to the kinetics of phosphorylation/dephosphorylation. The cellular response networks identified provide a template for further exploration of the mechanisms of trichothecenemycotoxins and other ribotoxins, and ultimately, could contribute to improved mechanism-based human health risk assessment. - Highlights: ► Mycotoxin deoxynivalenol (DON) induces immunotoxicity via ribotoxic stress response. ► SILAC phosphoproteomics using TiO{sub 2} was applied to DON-treated RAW 264.7 cells. ► DON induces extensive protein phosphorylation changes involving 188 phosphoproteins. ► The main target of early DON-induced RSR is transcriptional regulation. ► Early DON-induced RSR is mediated by MAPK-, NFκB-, AKT- and AMPK-linked pathways.« less
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De Zutter, N; Audenaert, K; Arroyo-Manzanares, N; De Boevre, M; Van Poucke, C; De Saeger, S; Haesaert, G; Smagghe, G
2016-12-08
Biotransformation of mycotoxins in animals comprises phase I and phase II metabolisation reactions. For the trichothecene deoxynivalenol (DON), several phase II biotransformation reactions have been described resulting in DON-glutathiones, DON-glucuronides and DON-sulfates made by glutathione-S-transferases, uridine-diphosphoglucuronyl transferases and sulfotransferases, respectively. These metabolites can be easily excreted and are less toxic than their free compounds. Here, we demonstrate for the first time in the animal kingdom the conversion of DON to DON-3-glucoside (DON-3G) via a model system with plant pathogenic aphids. This phase II biotransformation mechanism has only been reported in plants. As the DON-3G metabolite was less toxic for aphids than DON, this conversion is considered a detoxification reaction. Remarkably, English grain aphids (Sitobion avenae) which co-occur with the DON producer Fusarium graminearum on wheat during the development of fusarium symptoms, tolerate DON much better and convert DON to DON-3G more efficiently than pea aphids (Acyrthosiphon pisum), the latter being known to feed on legumes which are no host for F. graminearum. Using a non-targeted high resolution mass spectrometric approach, we detected DON-diglucosides in aphids probably as a result of sequential glucosylation reactions. Data are discussed in the light of an eventual co-evolutionary adaptation of S. avenae to DON.
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De Zutter, N.; Audenaert, K.; Arroyo-Manzanares, N.; De Boevre, M.; Van Poucke, C.; De Saeger, S.; Haesaert, G.; Smagghe, G.
2016-01-01
Biotransformation of mycotoxins in animals comprises phase I and phase II metabolisation reactions. For the trichothecene deoxynivalenol (DON), several phase II biotransformation reactions have been described resulting in DON-glutathiones, DON-glucuronides and DON-sulfates made by glutathione-S-transferases, uridine-diphosphoglucuronyl transferases and sulfotransferases, respectively. These metabolites can be easily excreted and are less toxic than their free compounds. Here, we demonstrate for the first time in the animal kingdom the conversion of DON to DON-3-glucoside (DON-3G) via a model system with plant pathogenic aphids. This phase II biotransformation mechanism has only been reported in plants. As the DON-3G metabolite was less toxic for aphids than DON, this conversion is considered a detoxification reaction. Remarkably, English grain aphids (Sitobion avenae) which co-occur with the DON producer Fusarium graminearum on wheat during the development of fusarium symptoms, tolerate DON much better and convert DON to DON-3G more efficiently than pea aphids (Acyrthosiphon pisum), the latter being known to feed on legumes which are no host for F. graminearum. Using a non-targeted high resolution mass spectrometric approach, we detected DON-diglucosides in aphids probably as a result of sequential glucosylation reactions. Data are discussed in the light of an eventual co-evolutionary adaptation of S. avenae to DON. PMID:27929076
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Serum cation profile of broilers at various stages of exposure to deoxynivalenol.
Yunus, Agha Waqar; Böhm, Josef
2013-05-01
The present experiment was carried out to investigate if levels of serum cations in broilers are modulated differently at various stages of exposure to deoxynivalenol (DON). Male broiler chicks at 7 days of age were fed a basal diet (0.27 mg of DON; 0.01 mg of zearalenone/kg), or either a low DON diet (1.68 mg of DON; 0.15 mg of zearalenone/kg) or a high DON diet (12.21 mg of DON; 1.09 mg of zearalenone/kg) produced using extracts from Fusarium graminearum cultures. Blood samples from the birds were collected during weeks 2, 4, and 5 of exposure. The high DON diet resulted in lower serum calcium levels compared to the basal diet at all the 3 sampling stages, while the low DON diet resulted in lower serum calcium levels only during weeks 2 and 5. Serum potassium levels were reduced under both the DON diets during weeks 2 and 5, while no diet-associated changes were found for serum levels of magnesium, sodium, and zinc. Under the present experimental conditions, the serum levels of calcium were consistently modulated in the broilers exposed to the DON-contaminated diets. The modulation of serum levels of potassium was, however, dependent upon the stage of exposure to DON.
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Kluger, Bernhard; Bueschl, Christoph; Lemmens, Marc; Michlmayr, Herbert; Malachova, Alexandra; Koutnik, Andrea; Maloku, Imer; Berthiller, Franz; Adam, Gerhard; Krska, Rudolf; Schuhmacher, Rainer
2015-01-01
In this study, a total of nine different biotransformation products of the Fusarium mycotoxin deoxynivalenol (DON) formed in wheat during detoxification of the toxin are characterized by liquid chromatography—high resolution mass spectrometry (LC-HRMS). The detected metabolites suggest that DON is conjugated to endogenous metabolites via two major metabolism routes, namely 1) glucosylation (DON-3-glucoside, DON-di-hexoside, 15-acetyl-DON-3-glucoside, DON-malonylglucoside) and 2) glutathione conjugation (DON-S-glutathione, “DON-2H”-S-glutathione, DON-S-cysteinyl-glycine and DON-S-cysteine). Furthermore, conjugation of DON to a putative sugar alcohol (hexitol) was found. A molar mass balance for the cultivar ‘Remus’ treated with 1 mg DON revealed that under the test conditions approximately 15% of the added DON were transformed into DON-3-glucoside and another 19% were transformed to the remaining eight biotransformation products or irreversibly bound to the plant matrix. Additionally, metabolite abundance was monitored as a function of time for each DON derivative and was established for six DON treated wheat lines (1 mg/ear) differing in resistance quantitative trait loci (QTL) Fhb1 and/or Qfhs.ifa-5A. All cultivars carrying QTL Fhb1 showed similar metabolism kinetics: Formation of DON-Glc was faster, while DON-GSH production was less efficient compared to cultivars which lacked the resistance QTL Fhb1. Moreover, all wheat lines harboring Fhb1 showed significantly elevated D3G/DON abundance ratios. PMID:25775425
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Huang, Juan; Chen, Guosong; Zhang, Xiaoyan; Shen, Chongyu; Lü, Chen; Wu, Bin; Liu, Yan; Chen, Huilan; Ding, Tao
2012-11-01
A method was established for the determination of deoxynivalenol (vomitoxin) in grain and its products based on solid-phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The sample was firstly extracted by acetonitrile-water (84:16, v/v). The extract was then cleaned-up by an HLB solid phase extraction cartridge. The separation was carried out on a Phenomenex Kinetex C18 column (100 mm x4. 6 mm, 2.6 microm) with a gradient elution using 0.3% per hundred ammonia solution-acetonitrile as mobile phases. The analysis of deoxynivalenol was performed under electrospray negative ionization mode. The limit of detection (LOD, S/N= 3) and the limit of quantification (LOQ, S/N = 10) were 20 microg/kg and 50 microg/kg, respectively. A good linearity (r > 0.99) was achieved for the target compound over the range of 20-1000 pg/L. The recoveries at the three spiked levels (50, 100, 500 microg/kg) in the blank matrices such as flour, barley, soybean, rice, cornmeal, cassava and wheat, were varied from 75.6% to 111.0% with the relative standard deviations no more than 13. 0%. The method is accurate, efficient, sensitive and practical. The cost of pretreatment is obviously reduced by replacing immunoaffinity columns and Mycosep columns with HLB columns which have the same purification effect.
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Stability of DON and DON-3-glucoside during baking as affected by the presence of food additives.
Vidal, Arnau; Sanchis, Vicente; Ramos, Antonio J; Marín, Sonia
2018-03-01
The mycotoxin deoxynivalenol (DON) is one of the most common mycotoxins of cereals worldwide, and its occurrence has been widely reported in raw wheat. The free mycotoxin form is not the only route of exposure; modified forms can also be present in cereal products. Deoxynivalenol-3-glucoside (DON-3-glucoside) is a common DON plant conjugate. The mycotoxin concentration could be affected by food processing; here, we studied the stability of DON and DON-3-glucoside during baking of small doughs made from white wheat flour and other ingredients. A range of common food additives and ingredients were added to assess possible interference: ascorbic acid (E300), citric acid (E330), sorbic acid (E200), calcium propionate (E282), lecithin (E322), diacetyltartaric acid esters of fatty acid mono- and diglycerides (E472a), calcium phosphate (E341), disodium diphosphate (E450i), xanthan gum (E415), polydextrose (E1200), sorbitol (E420i), sodium bicarbonate (E500i), wheat gluten and malt flour. The DON content was reduced by 40%, and the DON-3-glucoside concentration increased by >100%, after baking for 20 min at 180°C. This confirmed that DON and DON-3-glucoside concentrations can vary during heating, and DON-3-glucoside could even increase after baking. However, DON and DON-3-glucoside are not affected significantly by the presence of the food additives tested.
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USDA-ARS?s Scientific Manuscript database
Fusarium head blight (FHB), mainly caused by Fusarium graminearum, is a devastating disease of wheat and barley that results in huge economic losses worldwide. During infection, F. graminearum produces trichothecene mycotoxins, such as deoxynivalenol (DON), that increase fungal virulence and decreas...
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Warth, Benedikt; Del Favero, Giorgia; Wiesenberger, Gerlinde; Puntscher, Hannes; Woelflingseder, Lydia; Fruhmann, Philipp; Sarkanj, Bojan; Krska, Rudolf; Schuhmacher, Rainer; Adam, Gerhard; Marko, Doris
2016-01-01
The mycotoxin deoxynivalenol (DON) is an abundant contaminant of cereal based food and a severe issue for global food safety. We report the discovery of DON-3-sulfate as a novel human metabolite and potential new biomarker of DON exposure. The conjugate was detectable in 70% of urine samples obtained from pregnant women in Croatia. For the measurement of urinary metabolites, a highly sensitive and selective LC-MS/MS method was developed and validated. The method was also used to investigate samples from a duplicate diet survey for studying the toxicokinetics of DON-3-sulfate. To get a preliminary insight into the biological relevance of the newly discovered DON-sulfates, in vitroexperiments were performed. In contrast to DON, sulfate conjugates lacked potency to suppress protein translation. However, surprisingly we found that DON-sulfates enhanced proliferation of human HT-29 colon carcinoma cells, primary human colon epithelial cells (HCEC-1CT) and, to some extent, also T24 bladder cancer cells. A proliferative stimulus, especially in tumorigenic cells raises concern on the potential impact of DON-sulfates on consumer health. Thus, a further characterization of their toxicological relevance should be of high priority. PMID:27659167
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NASA Astrophysics Data System (ADS)
Warth, Benedikt; Del Favero, Giorgia; Wiesenberger, Gerlinde; Puntscher, Hannes; Woelflingseder, Lydia; Fruhmann, Philipp; Sarkanj, Bojan; Krska, Rudolf; Schuhmacher, Rainer; Adam, Gerhard; Marko, Doris
2016-09-01
The mycotoxin deoxynivalenol (DON) is an abundant contaminant of cereal based food and a severe issue for global food safety. We report the discovery of DON-3-sulfate as a novel human metabolite and potential new biomarker of DON exposure. The conjugate was detectable in 70% of urine samples obtained from pregnant women in Croatia. For the measurement of urinary metabolites, a highly sensitive and selective LC-MS/MS method was developed and validated. The method was also used to investigate samples from a duplicate diet survey for studying the toxicokinetics of DON-3-sulfate. To get a preliminary insight into the biological relevance of the newly discovered DON-sulfates, in vitroexperiments were performed. In contrast to DON, sulfate conjugates lacked potency to suppress protein translation. However, surprisingly we found that DON-sulfates enhanced proliferation of human HT-29 colon carcinoma cells, primary human colon epithelial cells (HCEC-1CT) and, to some extent, also T24 bladder cancer cells. A proliferative stimulus, especially in tumorigenic cells raises concern on the potential impact of DON-sulfates on consumer health. Thus, a further characterization of their toxicological relevance should be of high priority.
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Effect of cleaning, milling, and baking on deoxynivalenol in wheat.
Abbas, H K; Mirocha, C J; Pawlosky, R J; Pusch, D J
1985-01-01
Samples of wheat naturally infected by Fusarium graminearum Schwabe were obtained from mills in Oklahoma, Missouri, Kansas, and Minnesota and fields in Nebraska and Kansas in 1982; they were analyzed for deoxynivalenol (DON). The wheat was milled, and DON was found throughout all the milling fractions (bran, shorts, reduction flour, and break flour). The DON recoveries for each mill run ranged from 90 to 98%. These samples, regardless of DON concentration, also gave similar fractional distributions of DON. The greatest (21 ppm [21 micrograms/g]) concentration of DON was found in the bran, and the smallest (1 ppm) was found in the break flour. Cleaning and milling were not effective in removing DON; DON was not destroyed in the bread baked from the naturally contaminated whole wheat flour, but the effect on its concentration in the samples analyzed varied, the reduction ranging from 19 to 69%. The percent reduction found in the cleaned wheat ranged from 6 to 19%. DON concentrations in the following commercially made breads, caraway rye, seedless rye, and pumpernickel, were 45 ppb (ng/g), 39 ppb, and 0 ppb, respectively. The limits of detection by gas chromatography-mass spectrometry and high-pressure liquid chromatography for DON were 0.5 and 10 ng, respectively. PMID:4051489
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USDA-ARS?s Scientific Manuscript database
Nivalenol (NIV) and Deoxynivalenol (DON) are trichothecene mycotoxins produced by Fusarium spp. that contaminate mainly cereal crops, such as wheat, barley, and maize. These mycotoxins are serious health hazards to humans and domestic animals. In Japan, there have been many reports of DON and NIV ...
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Effect of Ozone Treatment on Deoxynivalenol and Wheat Quality
Wang, Li; Shao, Huili; Luo, Xiaohu; Wang, Ren; Li, Yongfu; Li, Yanan; Luo, Yingpeng; Chen, Zhengxing
2016-01-01
Deoxynivalenol (DON) is a secondary metabolite produced by Fusarium fungi, which is found in a wide range of agricultural products, especially in wheat, barley, oat and corn. In this study, the distribution of DON in the wheat kernel and the effect of exposure time to ozone on DON detoxification were investigated. A high concentration of toxin was found in the outer part of the kernel, and DON was injected from the outside to the inside. The degradation rates of DON were 26.40%, 39.16%, and 53.48% after the samples were exposed to 75 mg/L ozone for 30, 60, and 90 min, respectively. The effect of ozonation on wheat flour quality and nutrition was also evaluated. No significant differences (P > 0.05) were found in protein content, fatty acid value, amino acid content, starch content, carbonyl and carboxyl content, and swelling power of ozone-treated samples. Moreover, the ozone-treated samples exhibited higher tenacity and whiteness, as well as lower extensibility and yellowness. This finding indicated that ozone treatment can simultaneously reduce DON levels and improve flour quality. PMID:26812055
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Li, Ruonan; Li, Yansen; Su, Yongteng; Shen, Dan; Dai, Pengyuan; Li, Chunmei
2018-05-28
The mycotoxin deoxynivalenol (DON) generally exists in cereals and affects human and animal health. The aim of this study is to analyze the impacts of DON in naturally contaminated feed on piglet growth performance and intestinal hormone secretion in the short term. We randomly divided 5-week-old piglets into four groups: Control, DON 1,000, DON 2,000 and DON 3,000 groups. Piglets received a feed naturally contaminated with DON (approximately 400, 1,000, 2,000 or 3,000 μg/kg) for 21 days. Body weight showed no significant difference following exposure to DON. The balance of anti-oxidation and oxidation was disrupted by DON after 21 days. The concentration of tumor necrosis factor-alpha (TNF-α) and cyclooxgenase-2 (COX-2) significantly increased (p < .001) in all DON-treated groups. Gut anorexigenic hormone secretion of peptide YY (PYY) and cholecystokinin (CCK) had a time- and dose-dependent relationship with DON exposure; however, there was no effect on orexigenic hormone ghrelin secretion. Changes of histomorphology in the jejunum were observed in DON-treated groups, including villi flattening and fusion, and apical necrosis of villi. These results indicated that DON could suppress piglet growth performance and alter gut hormone secretion in the short term. © 2018 Japanese Society of Animal Science.
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Vidal, A; Marín, S; Morales, H; Ramos, A J; Sanchis, V
2014-06-01
Deoxynivalenol (DON) and ochratoxin A (OTA) are mycotoxins produced by fungal species which can contaminate, alone or simultaneously, cereal-based products such as bread. Due to the increasing interest in the beneficial effects of dietary bran, bran bread has attained high consumption. Usually, the higher mycotoxin concentrations in cereals are found in the external layers of the grain (bran), leading to higher concentration of DON and OTA in breads with added bran. Moreover, the use of sourdough in breadmaking is increasing, but no studies about its effect in the mycotoxins content exist. The objective of this study was to determine the variation of concentration of these mycotoxins during the breadmaking process including the following factors: two initial mycotoxin concentrations in the initial mix of ingredients, four different bran contents, and use of sourdough. OTA was confirmed to be quite stable during the breadmaking process, regardless of the assayed factors. DON concentration during breadmaking was not significantly affected by bran content of bread. However, it was significantly affected by kneading and fermentation steps in opposite way depending on sourdough use and flour contamination level: if DON reduction occurs during fermentation, this leads to a safer situation, but the possible increase in DON should be considered with care, as it can compensate the expected dilution effect by recipe. Finally, the results on deoxynivalenol-3-glucoside (DON-3-G), although preliminar, suggest an increase of this toxin during fermentation, but mainly during baking. Copyright © 2014 Elsevier Ltd. All rights reserved.
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USDA-ARS?s Scientific Manuscript database
Current models to forecast Fusarium head blight (FHB) and deoxynivalenol (DON) levels in wheat are based on weather near anthesis, and breeding for resistance to Fusarium often relies on irrigation before and shortly after anthesis to encourage disease development. The effects of post-anthesis envi...
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USDA-ARS?s Scientific Manuscript database
Fusarium Head Blight (FHB) is a disease caused by the fungal pathogen Fusarium graminearum that affects wheat and other small grain cereals and can lead to severe yield loss and reduction in grain quality. Trichothecene mycotoxins, such as deoxynivalenol (DON), accumulate during infection and increa...
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USDA-ARS?s Scientific Manuscript database
Fusarium head blight (FHB) or scab, caused by Fusarium graminearum Schwabe [sexual stage Gibberella zeae (Schwein.) Petch], is a destructive disease of small grain cereals. Fusarium graminearum produces the mycotoxin deoxynivalenol (DON), which accumulates in and lowers the value and quality of grai...
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USDA-ARS?s Scientific Manuscript database
Nivalenol (NIV) and Deoxynivalenol (DON) are trichothecene mycotoxins produced by Fusarium spp that contaminate mainly cereal crops, such as wheat, barley, and maize. These mycotoxins are serious health hazards to human and domestic animals. The study reports a rapid screening method of NIV and DO...
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Sanders, Melanie; Guo, Yirong; Iyer, Abhishek; García, Yara Ruiz; Galvita, Anastasia; Heyerick, Arne; Deforce, Dieter; Risseeuw, Martijn D P; Van Calenbergh, Serge; Bracke, Marc; Eremin, Sergei; Madder, Annemieke; De Saeger, Sarah
2014-01-01
An immunogen synthesis strategy was designed to develop anti-deoxynivalenol (DON) monoclonal antibodies with low cross-reactivity against structurally similar trichothecenes. A total of eight different DON immunogens were synthesised, differing in the type and position of the linker on the DON molecule. After immunisation, antisera from mice immunised with different DON immunogens were checked for the presence of relevant antibodies. Then, both homologous and heterologous enzyme-linked immunosorbent assays (ELISAs) were performed for hybridoma screening. Finally, three monoclonal antibodies against DON and its analogues were generated. In addition, monoclonal antibody 13H1 could recognise DON and its analogues in the order of HT-2 toxin > 15-acetyldeoxynivalenol (15-ADON) > DON, with IC₅₀ ranging from 1.14 to 2.13 µg ml⁻¹. Another monoclonal antibody 10H10 manifested relatively close sensitivities to DON, 3-acetyldeoxynivalenol (3-ADON) and 15-ADON, with IC₅₀ values of 22, 15 and 34 ng ml⁻¹, respectively. Using an indirect ELISA format decreases the 10H10 sensitivity to 15-ADON with 92%. A third monoclonal antibody 2A9 showed to be very specific and sensitive to 3-ADON, with IC₅₀ of 0.38 ng ml⁻¹. Using both 2A9 and 10H10 monoclonal antibodies allows determining sole DON contamination.
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Michlmayr, Herbert; Malachová, Alexandra; Varga, Elisabeth; Kleinová, Jana; Lemmens, Marc; Newmister, Sean; Rayment, Ivan; Berthiller, Franz; Adam, Gerhard
2015-01-01
Glycosylation is an important plant defense mechanism and conjugates of Fusarium mycotoxins often co-occur with their parent compounds in cereal-based food and feed. In case of deoxynivalenol (DON), deoxynivalenol-3-O-β-d-glucoside (D3G) is the most important masked mycotoxin. The toxicological significance of D3G is not yet fully understood so that it is crucial to obtain this compound in pure and sufficient quantities for toxicological risk assessment and for use as an analytical standard. The aim of this study was the biochemical characterization of a DON-inactivating UDP-glucosyltransferase from rice (OsUGT79) and to investigate its suitability for preparative D3G synthesis. Apparent Michaelis constants (Km) of recombinant OsUGT79 were 0.23 mM DON and 2.2 mM UDP-glucose. Substrate inhibition occurred at DON concentrations above 2 mM (Ki = 24 mM DON), and UDP strongly inhibited the enzyme. Cu2+ and Zn2+ (1 mM) inhibited the enzyme completely. Sucrose synthase AtSUS1 was employed to regenerate UDP-glucose during the glucosylation reaction. With this approach, optimal conversion rates can be obtained at limited concentrations of the costly co-factor UDP-glucose. D3G can now be synthesized in sufficient quantity and purity. Similar strategies may be of interest to produce β-glucosides of other toxins. PMID:26197338
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Dänicke, Sven; Beineke, Andreas; Berk, Andreas; Kersten, Susanne
2017-01-01
The common feed contaminant deoxynivalenol (DON) was reported to influence the morphology of the pars nonglandularis (PN) of porcine stomach. Moreover, finely ground feed is known to trigger the development of ulcers and other pathologies of PN while coarsely ground feed protects from such lesions. The interactions between grinding fineness and DON contamination of feed were not examined so far. Therefore, both finely and coarsely ground feeds were tested either in the absence or presence of a DON contaminated wheat on growth performance and health of rearing piglets, including stomach integrity. DON contamination significantly reduced feed intake and serum albumin concentration with this effect being more pronounced after feeding the coarsely ground feed. Albeit at a higher level, albumin concentration was also reduced after feeding the finely ground and uncontaminated feed. Finely ground and DON-contaminated feed caused a significantly more pronounced lymphoplasmacytic infiltration both of PN and pars glandularis , partly paralleled by lymph follicle formation and detritus filled foveolae and tubes suggesting a local immune response probably triggered by epithelial lesions. It is concluded that DON contamination of feed exacerbates the adverse effects of finely ground feed on stomach mucosal integrity.
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Dänicke, Sven; Beineke, Andreas; Berk, Andreas; Kersten, Susanne
2017-01-01
The common feed contaminant deoxynivalenol (DON) was reported to influence the morphology of the pars nonglandularis (PN) of porcine stomach. Moreover, finely ground feed is known to trigger the development of ulcers and other pathologies of PN while coarsely ground feed protects from such lesions. The interactions between grinding fineness and DON contamination of feed were not examined so far. Therefore, both finely and coarsely ground feeds were tested either in the absence or presence of a DON contaminated wheat on growth performance and health of rearing piglets, including stomach integrity. DON contamination significantly reduced feed intake and serum albumin concentration with this effect being more pronounced after feeding the coarsely ground feed. Albeit at a higher level, albumin concentration was also reduced after feeding the finely ground and uncontaminated feed. Finely ground and DON-contaminated feed caused a significantly more pronounced lymphoplasmacytic infiltration both of PN and pars glandularis, partly paralleled by lymph follicle formation and detritus filled foveolae and tubes suggesting a local immune response probably triggered by epithelial lesions. It is concluded that DON contamination of feed exacerbates the adverse effects of finely ground feed on stomach mucosal integrity. PMID:28045426
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Li, Xin; Shin, Sanghyun; Heinen, Shane; Dill-Macky, Ruth; Berthiller, Franz; Nersesian, Natalya; Clemente, Thomas; McCormick, Susan; Muehlbauer, Gary J
2015-11-01
Fusarium head blight (FHB), mainly caused by Fusarium graminearum, is a devastating disease of wheat that results in economic losses worldwide. During infection, F. graminearum produces trichothecene mycotoxins, including deoxynivalenol (DON), that increase fungal virulence and reduce grain quality. Transgenic wheat expressing a barley UDP-glucosyltransferase (HvUGT13248) were developed and evaluated for FHB resistance, DON accumulation, and the ability to metabolize DON to the less toxic DON-3-O-glucoside (D3G). Point-inoculation tests in the greenhouse showed that transgenic wheat carrying HvUGT13248 exhibited significantly higher resistance to disease spread in the spike (type II resistance) compared with nontransformed controls. Two transgenic events displayed complete suppression of disease spread in the spikes. Expression of HvUGT13248 in transgenic wheat rapidly and efficiently conjugated DON to D3G, suggesting that the enzymatic rate of DON detoxification translates to type II resistance. Under field conditions, FHB severity was variable; nonetheless, transgenic events showed significantly less-severe disease phenotypes compared with the nontransformed controls. In addition, a seedling assay demonstrated that the transformed plants had a higher tolerance to DON-inhibited root growth than nontransformed plants. These results demonstrate the utility of detoxifying DON as a FHB control strategy in wheat.
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Reddy, Kondreddy Eswar; Song, Jaeyong; Lee, Hyun-Jeong; Kim, Minseok; Kim, Dong-Wook; Jung, Hyun Jung; Kim, Bumseok; Lee, Yookyung; Yu, Dongjo; Kim, Dong-Woon; Oh, Young Kyoon; Lee, Sung Dae
2018-01-01
Background: Deoxynivalenol (DON) and zearalenone (ZEN) are common food contaminants produced by Fusarium sp. Mycotoxins are a potential health hazard because of their toxicological effects on both humans and farmed animals. Methods: We analyzed three groups of pigs: a control group (fed a standard diet), and the DON and ZEN groups, fed a diet containing 8 mg/kg DON and 0.8 mg/kg ZEN respectively, for four weeks. Results: DON and ZEN exposure decreased body weight (BW), average daily feed intake (ADFI), food conversion rate (FCR), and the serum levels of immunoglobulin (Ig)G and IgM. The total antioxidant levels significantly decreased in serum and increased in urine samples of both treatment groups. Additionally, DON and ZEN exposure increased serotonin levels in urine. Hematological parameters were not affected by the investigated toxins. Microscopic lesions were evident in sections of kidneys from either treatment group: we found sporadic interstitial nephritis in the DON group and renal glomerulus atrophy in the ZEN group. The expression levels of inflammatory cytokines and chemokine marker genes were reduced in tissues from DON- and ZEN-exposed pigs. Conclusions: chronic ingestion of high doses of DON and ZEN alters the immune response and causes organs damage, and might be associated with various diseases in pigs. PMID:29518941
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Effects of the mycotoxin deoxynivalenol on steroidogenesis and apoptosis in granulosa cells.
Guerrero-Netro, Hilda M; Chorfi, Younès; Price, Christopher A
2015-06-01
Mycotoxins can reduce fertility and development in livestock, notably in pigs and poultry, although the effect of most mycotoxins on reproductive function in cattle has not been established. One major mycotoxin, deoxynivalenol (DON), not only targets immune cells and activates the ribotoxic stress response (RSR) involving MAPK activation, but also inhibits oocyte maturation in pigs. In this study, we determined the effect of DON on bovine granulosa cell function using a serum-free culture system. Addition of DON inhibited estradiol and progesterone secretion, and reduced levels of mRNA encoding estrogenic (CYP19A1) but not progestogenic (CYP11A1 and STAR) proteins. Cell apoptosis was increased by DON, which also increased FASLG mRNA levels. The mechanism of action of DON was assessed by western blotting and PCR experiments. Addition of DON rapidly and transiently increased phosphorylation of MAPK3/1, and resulted in a more prolonged phosphorylation of MAPK14 (p38) and MAPK8 (JNK). Activation of these pathways by DON resulted in time- and dose-dependent increases in abundance of mRNA encoding the transcription factors FOS, FOSL1, EGR1, and EGR3. We conclude that DON is deleterious to granulosa cell function and acts through a RSR pathway. © 2015 Society for Reproduction and Fertility.
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USDA-ARS?s Scientific Manuscript database
Fusarium head blight is a plant disease with significant agricultural and health impact which affects cereal crops such as wheat, barley, and maize and is characterized by reduced grain yield and the accumulation of trichothecene mycotoxins such as deoxynivalenol (DON). Studies have identified trich...
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USDA-ARS?s Scientific Manuscript database
Resistance in oats (Avena sativa L.) to infection by Fusarium graminearum was assessed in field trials in 2011-12 including 424 spring oat lines from North America and Scandinavia. Traits measured were Fusarium Head Blight (FHB), deoxynivalenol (DON) content, days to flowering (DTF) and days to matu...
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Ren, Zhihua; Wang, Yachao; Deng, Huidan; Deng, Youtian; Deng, Junliang; Zuo, Zhicai; Wang, Ya; Peng, Xi; Cui, Hengmin; Shen, Liuhong
2015-01-01
We investigated the immunotoxicity and cytotoxicity of deoxynivalenol (DON), a mycotoxin, and the mechanism by which it induces apoptosis. Chicken splenic lymphocytes treated with 0-50μg/mL DON for 48h inhibited growth of splenic lymphocytes in a dose-dependent manner, as revealed by the Cell Counting Kit-8 (CCK-8) bioassay. Annexin V-fluorescein isothiocyanate staining indicated that the number of apoptotic and necrotic cells were significantly higher compared with the control (P<0.01). DON treatment induced ROS accumulation, resulting in reduced mitochondrial transmembrane potential, as detected by flow cytometry and 2',7'-dichlorofluorescein acetate and rhodamine 123 labeling, respectively. Enzyme linked immunosorbent assays revealed that the concentrations of p53, Bax, Bak-1, and Caspase-3 increased with increasing DON concentration (P<0.05 or P<0.01), whereas the concentrations of Bcl-2 decreased (P<0.01) compared with the control. These data suggest that DON induces apoptosis in splenic lymphocytes via a ROS-mediated mitochondrial pathway. Copyright © 2014. Published by Elsevier B.V.
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Deoxynivalenol: A Major Player in the Multifaceted Response of Fusarium to Its Environment
Audenaert, Kris; Vanheule, Adriaan; Höfte, Monica; Haesaert, Geert
2013-01-01
The mycotoxin deoxynivalenol (DON), produced by several Fusarium spp., acts as a virulence factor and is essential for symptom development after initial wheat infection. Accumulating evidence shows that the production of this secondary metabolite can be triggered by diverse environmental and cellular signals, implying that it might have additional roles during the life cycle of the fungus. Here, we review data that position DON in the saprophytic fitness of Fusarium, in defense and in the primary C and N metabolism of the plant and the fungus. We combine the available information in speculative models on the role of DON throughout the interaction with the host, providing working hypotheses that await experimental validation. We also highlight the possible impact of control measures in the field on DON production and summarize the influence of abiotic factors during processing and storage of food and feed matrices. Altogether, we can conclude that DON is a very important compound for Fusarium to cope with a changing environment and to assure its growth, survival, and production of toxic metabolites in diverse situations. PMID:24451843
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Okuma, Tara A; Huynh, Thu P; Hellberg, Rosalee S
2018-03-01
The objective of this study was to perform a market survey on dry pet foods using enzyme-linked immunosorbent assay (ELISA) to detect total aflatoxins (AFs), ochratoxin A (OTA), and deoxynivalenol (DON). Pet food products (n = 58) marketed for dogs, cats, birds, and rabbits were tested in duplicate with ELISA, and results above the limit of quantitation were confirmed using liquid chromatography tandem mass spectrometry (LC-MS/MS). OTA was detected in one product (rabbit food) and AFs were detected in two products (one dog treat and one bird treat). In contrast, DON was detected in the majority (74%) of products tested. Bird and rabbit products were the most affected by DON, with levels above 0.5 μg/g in 50 and 80% of samples, respectively. One rabbit sample tested positive for both OTA and DON. Overall, the findings of this study revealed a low incidence of AFs and OTA in commercial pet food. Although DON was detected in numerous products, the levels were well below those associated with acute toxic effects.
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Occurrence of Deoxynivalenol and Zearalenone in Commercial Fish Feed: An Initial Study
Pietsch, Constanze; Kersten, Susanne; Burkhardt-Holm, Patricia; Valenta, Hana; Dänicke, Sven
2013-01-01
The control of mycotoxins is a global challenge not only in human consumption but also in nutrition of farm animals including aquatic species. Fusarium toxins, such as deoxynivalenol (DON) and zearalenone (ZEN), are common contaminants of animal feed but no study reported the occurrence of both mycotoxins in fish feed so far. Here, we report for the first time the occurrence of DON and ZEN in samples of commercial fish feed designed for nutrition of cyprinids collected from central Europe. A maximal DON concentration of 825 μg kg−1 feed was found in one feed whereas average values of 289 μg kg−1 feed were noted. ZEN was the more prevalent mycotoxin but the concentrations were lower showing an average level of 67.9 μg kg−1 feed. PMID:23325300
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Kos, Gregor; Lohninger, Hans; Mizaikoff, Boris; Krska, Rudolf
2007-07-01
A sample preparation procedure for the determination of deoxynivalenol (DON) using attenuated total reflection mid-infrared spectroscopy is presented. Repeatable spectra were obtained from samples featuring a narrow particle size distribution. Samples were ground with a centrifugal mill and analysed with an analytical sieve shaker. Particle sizes of <100, 100-250, 250-500, 500-710 and 710-1000 microm were obtained. Repeatability, classification and quantification abilities for DON were compared with non-sieved samples. The 100-250 microm fraction showed the best repeatability. The relative standard deviation of spectral measurements improved from 20 to 4.4% and 100% of sieved samples were correctly classified compared with 79% of non-sieved samples. The DON level in analysed fractions was a good estimate of overall toxin content.
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Pan, D; Mionetto, A; Tiscornia, S; Bettucci, L
2015-08-01
In Uruguay, Fusarium graminearum is the most common species that infects wheat and is responsible for Fusarium head blight (FHB) and contamination of grain with deoxynivalenol (DON). The aim of this work was to select bacterial endophytes isolated from wheat grain to evaluate their antagonistic ability against F. graminearum and DON production in vitro and under field conditions. Four strains identified as Bacillus megaterium (BM1) and Bacillus subtilis (BS43, BSM0 y BSM2) significantly reduced fungal growth and spore germination of F. graminearum. This antagonist activity remained unchanged after the bacterial cultures were heat treated. Under field conditions, treatments with antagonist BM1 was the most effective, reducing the FHB incidence and severity by 93 and 54 %, respectively, and the production of DON by 89.3 %.
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Thermal degradation of deoxynivalenol during maize bread baking.
Numanoglu, E; Gökmen, V; Uygun, U; Koksel, H
2012-01-01
The thermal degradation of deoxynivalenol (DON) was determined at isothermal baking conditions within the temperature range of 100-250°C, using a crust-like model, which was prepared with naturally contaminated maize flour. No degradation was observed at 100°C. For the temperatures of 150, 200 and 250°C, thermal degradation rate constants (k) were calculated and temperature dependence of DON degradation was observed by using Arrhenius equation. The degradation of DON obeyed Arrhenius law with a regression coefficient of 0.95. A classical bread baking operation was also performed at 250°C for 70 min and the rate of DON degradation in the bread was estimated by using the kinetic data derived from the model study. The crust and crumb temperatures recorded during bread baking were used to calculate the thermal degradation rate constants (k) and partial DON degradations at certain time intervals. Using these data, total degradation at the end of the entire baking process was predicted for both crust and crumb. This DON degradation was consistent with the experimental degradation data, confirming the accuracy of kinetic constants determined by means of the crust-like model.
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Preliminary Estimation of Deoxynivalenol Excretion through a 24 h Pilot Study
Rodríguez-Carrasco, Yelko; Mañes, Jordi; Berrada, Houda; Font, Guillermina
2015-01-01
A duplicate diet study was designed to explore the occurrence of 15 Fusarium mycotoxins in the 24 h-diet consumed by one volunteer as well as the levels of mycotoxins in his 24 h-collected urine. The employed methodology involved solvent extraction at high ionic strength followed by dispersive solid phase extraction and gas chromatography determination coupled to mass spectrometry in tandem. Satisfactory results in method performance were achieved. The method’s accuracy was in a range of 68%–108%, with intra-day relative standard deviation and inter-day relative standard deviation lower than 12% and 15%, respectively. The limits of quantitation ranged from 0.1 to 8 µg/Kg. The matrix effect was evaluated and matrix-matched calibrations were used for quantitation. Only deoxynivalenol (DON) was quantified in both food and urine samples. A total DON daily intake amounted to 49.2 ± 5.6 µg whereas DON daily excretion of 35.2 ± 4.3 µg was determined. DON daily intake represented 68.3% of the established DON provisional maximum tolerable daily intake (PMTDI). Valuable preliminary information was obtained as regards DON excretion and needs to be confirmed in large-scale monitoring studies. PMID:25723325
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He, Cheng-Hua; Fan, Yan-Hong; Wang, Ying; Huang, Chao-Ying; Wang, Xi-Chun; Zhang, Hai-Bin
2010-01-01
Aflatoxin B1 (AFB1) and deoxynivalenol (DON) are important food-borne mycotoxins that have been implicated in animal and human health. In this study, individual and combinative effects of AFB1 and DON were tested in primary hepatocytes of Cyprinus carpio. The results indicated that the combinative effects of AFB1 and DON (0.01 μg/mL AFB1 and 0.25 μg/mL DON; 0.02 μg/mL AFB1 and 0.25 μg/mL DON; 0.02 μg/mL AFB1 and 0.5 μg/mL DON) were higher than that of individual mycotoxin (P < 0.05). The activity of AST, ALT and LDH in cell supernatant was higher than that of control group (P < 0.05) when the mycotoxins were exposed to primary hepatocytes for 4 h. The decreased cell number was observed in tested group by inverted light microscopy. The mitochondrial swelling, endoplasmic reticulum dilation and a lot of lipid droplets were observed in primary hepatocytes by transmission electron microscope. Therefore, this combination was classified as an additive response of the two mycotoxins. PMID:21152299
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DON occurrence in cereal grains: A North American perspective
USDA-ARS?s Scientific Manuscript database
The occurrence of deoxynivalenol (DON) in agricultural commodities has been reported all over the world, with levels varying amongst grain type and years of production. However, a systematic review of the current situation regarding this mycotoxin, its occurrence, and its management in North America...
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Ren, Zhihua; Wang, Yachao; Deng, Huidan; Deng, Youtian; Deng, Junliang; Zuo, Zhicai; Wang, Ya; Peng, Xi; Cui, Hengmin; Shen, Liuhong; Yu, Shumin; Cao, Suizhong
2016-04-01
In this study, the in vitro effects of the treatment of concanavalin A (Con A)--stimulated splenic lymphocytes with DON were examined. Splenic lymphocytes isolated from chickens were stimulated with 12.5 μg/mL Con A and exposed to deoxynivalenol (DON) (0-50 μg/mL) for 48 h. The intracellular calcium concentration ([Ca(2+)]i), pH, calmodulin (CaM) mRNA levels, and Na(+),K(+)-ATPase and Ca(2+)-ATPase activities were detected. With the DON exposure concentrations increased, the [Ca(2+)]i and CaM mRNA levels gradually increased in a dose-dependent manner, and all the evaluated conconcentrations affected ATPase activity to the same extent. There were significant differences (P<0.05 or P<0.01) between the treatment groups and the control group. These results indicate that an imbalance in calcium homeostasis and intracellular acidification are components of DON cytotoxicity in chicken lymphocytes. Copyright © 2016 Elsevier GmbH. All rights reserved.
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Sanders, Melanie; McPartlin, Daniel; Moran, Kara; Guo, Yirong; Eeckhout, Mia; O'Kennedy, Richard; De Saeger, Sarah; Maragos, Chris
2016-04-11
A sample preparation method was developed for the screening of deoxynivalenol (DON) in wheat and wheat dust. Extraction was carried out with water and was successful due to the polar character of DON. For detection, an enzyme-linked immunosorbent assay (ELISA) was compared to the sensor-based techniques of surface plasmon resonance (SPR) and biolayer interferometry (BLI) in terms of sensitivity, affinity and matrix effect. The matrix effects from wheat and wheat dust using SPR were too high to further use this screenings method. The preferred ELISA and BLI methods were validated according to the criteria established in Commission Regulation 519/2014/EC and Commission Decision 2002/657/EC. A small survey was executed on 16 wheat lots and their corresponding dust samples using the validated ELISA method. A linear correlation (r = 0.889) was found for the DON concentration in dust versus the DON concentration in wheat (LOD wheat: 233 μg/kg, LOD wheat dust: 458 μg/kg).
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Antonissen, Gunther; Devreese, Mathias; Van Immerseel, Filip; De Baere, Siegrid; Hessenberger, Sabine; Martel, An; Croubels, Siska
2015-01-01
Both deoxynivalenol (DON) and fumonisin B1 (FB1) are common contaminants of feed. Fumonisins (FBs) in general have a very limited oral bioavailability in healthy animals. Previous studies have demonstrated that chronic exposure to DON impairs the intestinal barrier function and integrity, by affecting the intestinal surface area and function of the tight junctions. This might influence the oral bioavailability of FB1, and possibly lead to altered toxicity of this mycotoxin. A toxicokinetic study was performed with two groups of 6 broiler chickens, which were all administered an oral bolus of 2.5 mg FBs/kg BW after three-week exposure to either uncontaminated feed (group 1) or feed contaminated with 3.12 mg DON/kg feed (group 2). No significant differences in toxicokinetic parameters of FB1 could be demonstrated between the groups. Also, no increased or decreased body exposure to FB1 was observed, since the relative oral bioavailability of FB1 after chronic DON exposure was 92.2%. PMID:25690690
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Assessment of human deoxynivalenol exposure using an LC-MS/MS based biomarker method.
Warth, Benedikt; Sulyok, Michael; Fruhmann, Philipp; Berthiller, Franz; Schuhmacher, Rainer; Hametner, Christian; Adam, Gerhard; Fröhlich, Johannes; Krska, Rudolf
2012-05-20
The Fusarium toxin deoxynivalenol (DON) is one of the most abundant mycotoxins worldwide and poses many adverse health effects to human and animals. Consequently, regulatory limits and a provisional maximum tolerable daily intake (PMTDI) for this important type B-trichothecene were assigned. We conducted a pilot survey to investigate the level of DON exposure in Austrian adults by measurements of DON and its glucuronide conjugates (DON-GlcA's), as biomarkers of exposure, in first morning urine. The average concentration of total DON (free DON+DON-GlcA's) was estimated to be 20.4±2.4 μg L⁻¹ (max. 63 μg L⁻¹). Surprisingly, we found that one third of the volunteers (n=27) exceeded the established PMTDI when consuming regular diet. DON-GlcA's were directly quantified by LC-MS/MS and the results were compared with indirect quantification after enzymatic hydrolysis and confirmed the suitability of the direct method. Moreover, we investigated the in vivo metabolism of DON in humans and were able to determine two closely eluting DON-GlcA's in naturally contaminated urine samples for the first time. In contrast to previous findings we have tentatively identified DON-15-glucuronide as a major DON metabolite in human urine based on the analysis of these samples. About 75% of total glucuronides were derived from this metabolite while DON-3-glucuronide accounted for approximately 25%. The reported new findings clearly demonstrate the great potential of suitable biomarkers to critically assess exposure of humans and animals to DON. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.
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Varga, Elisabeth; Malachova, Alexandra; Nguyen, Nhung Thi; Lorenz, Cindy; Haltrich, Dietmar; Berthiller, Franz; Adam, Gerhard
2015-01-01
Glycosylation plays a central role in plant defense against xenobiotics, including mycotoxins. Glucoconjugates of Fusarium toxins, such as deoxynivalenol-3-O-β-d-glucoside (DON-3G), often cooccur with their parental toxins in cereal-based food and feed. To date, only limited information exists on the occurrence of glucosylated mycotoxins and their toxicological relevance. Due to a lack of analytical standards and the requirement of high-end analytical instrumentation for their direct determination, hydrolytic cleavage of β-glucosides followed by analysis of the released parental toxins has been proposed as an indirect determination approach. This study compares the abilities of several fungal and recombinant bacterial β-glucosidases to hydrolyze the model analyte DON-3G. Furthermore, substrate specificities of two fungal and two bacterial (Lactobacillus brevis and Bifidobacterium adolescentis) glycoside hydrolase family 3 β-glucosidases were evaluated on a broader range of substrates. The purified recombinant enzyme from B. adolescentis (BaBgl) displayed high flexibility in substrate specificity and exerted the highest hydrolytic activity toward 3-O-β-d-glucosides of the trichothecenes deoxynivalenol (DON), nivalenol, and HT-2 toxin. A Km of 5.4 mM and a Vmax of 16 μmol min−1 mg−1 were determined with DON-3G. Due to low product inhibition (DON and glucose) and sufficient activity in several extracts of cereal matrices, this enzyme has the potential to be used for indirect analyses of trichothecene-β-glucosides in cereal samples. PMID:25979885
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USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON) and fumonisins (FB) are the most frequently encountered mycotoxins produced by Fusarium species in livestock diets. The effect of subclinical doses of mycotoxins in chickens is largely unknown, and in particular the susceptibility of birds to pathogenic challenge when fed these ...
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Alizadeh, Arash; Braber, Saskia; Akbari, Peyman; Garssen, Johan; Fink-Gremmels, Johanna
2015-01-01
Deoxynivalenol (DON) is one of the major mycotoxins produced by Fusarium fungi, and exposure to this mycotoxin requires an assessment of the potential adverse effects, even at low toxin levels. The aim of this study was to investigate the effects of a short-term, low-dose DON exposure on various gut health parameters in pigs. Piglets received a commercial feed or the same feed contaminated with DON (0.9 mg/kg feed) for 10 days, and two hours after a DON bolus (0.28 mg/kg BW), weight gain was determined and samples of different segments of the intestine were collected. Even the selected low dose of DON in the diet negatively affected weight gain and induced histomorphological alterations in the duodenum and jejunum. The mRNA expression of different tight junction (TJ) proteins, especially occludin, of inflammatory markers, like interleukin-1 beta and interleukin-10 and the oxidative stress marker heme-oxigenase1, were affected along the intestine by low levels of DON in the diet. Taken together, our results indicate that even after low-level exposure to DON, which has been generally considered as acceptable in animal feeds, clinically-relevant changes are measurable in markers of gut health and integrity. PMID:26067367
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Chen, Stephanie S.; Li, Yi-Hung; Lin, Mei-Fong
2017-01-01
This study investigates the long-term effects of deoxynivalenol (DON) consumption on avian growth performance, on the proliferation, apoptosis, and DNA damage of spleen cells, and on intestinal integrity. Two hundred and eight 5-day-old black-feathered Taiwan country chickens were fed diets containing 0, 2, 5, and 10 mg/kg of DON for 16 weeks. Body weight gain of male birds in the 2 mg/kg group was significantly lower than that in the 5 mg/kg group. At the end of trial, feeding DON-contaminated diets of 5 mg/kg resulted in heavier spleens. Moreover, the increase in DON induced cellular proliferation, apoptosis, and DNA damage signals in the spleen, the exception being female birds fed 10 mg/kg of DON showing reduced proliferation. Expression of claudin-5 was increased in jejunum of female birds fed 2 and 5 mg/kg of DON, whereas decreased expression levels were found in male birds. In conclusion, our results verified that DON may cause a disturbance to the immune system and alter the intestinal barrier in Taiwan country chickens, and may also lead to discrepancies in growth performances in a dose- and sex-dependent manner. PMID:29053594
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The fungal myosin I is essential for Fusarium toxisome formation
USDA-ARS?s Scientific Manuscript database
The mycotoxin deoxynivalenol (DON) is the most frequently detected secondary metabolite produced by Fusarium graminearum and other Fusarium spp. To date, relatively few studies have addressed how mycotoxin biosynthesis occurs in fungal cells. Here we found that myosin I governs translation of DON bi...
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Manda, Gina; Mocanu, Mihaela Andreea; Marin, Daniela Eliza; Taranu, Ionelia
2015-02-16
Contamination of crops used for food and feed production with Fusarium mycotoxins, such as deoxynivalenol (DON), raise important health and economic issues all along the food chain. Acute exposure to high DON concentrations can alter the intestinal barrier, while chronic exposure to lower doses may exert more subtle effects on signal transduction pathways, leading to disturbances in cellular homeostasis. Using real-time cellular impedance measurements, we studied the effects exerted in vitro by low concentrations of DON (0.37-1.50 μM), relevant for mycotoxin-contaminated food, on the proliferation of undifferentiated Caco-2 cells presenting a tumorigenic phenotype. A 1.5 μM concentration of DON maintained cell adherence of non-proliferating Caco-2 cells, whilst arresting the growth of actively proliferating cells compared with control Caco-2 cells in vitro. At 0.37 μM, DON enhanced Caco-2 cell metabolism, thereby triggering a moderate increase in cell proliferation. The results of the current study suggested that low concentrations of DON commonly detected in food may either limit or sustain the proliferation of colon cancer cells, depending on their proliferation status and on DON concentration. Soluble factors released by Lactobacillus strains can partially counteract the inhibitory action of DON on actively proliferating colon cancer cells. The study also emphasized that real-time cellular impedance measurements were a valuable tool for investigating the dynamics of cellular responses to xenobiotics.
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Chang, Chao; Wang, Kun; Zhou, Sheng-Nan; Wang, Xue-Dong; Wu, Jin-E
2017-05-01
The aims of our study were to evaluate the effects of Saccharomyces boulardii (S. boulardii) on deoxynivalenol (DON)-induced injury in porcine alveolar macrophage cells (PAMCs) and to explore the underlying mechanisms. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometric analysis, ELISA, qRT-PCR, and western blot were performed to assess whether S. boulardii could prevent DON-induced injury by p38 mitogen-activated protein kinase (p38 MAPK) signal pathway. The results showed that pretreatment with 8 μM DON could decrease the viability of PAMC and significantly increase the apoptosis rate of PAMC, whereas S. boulardii could rescue apoptotic PAMC cells induced by DON. Further experiments revealed that S. boulardii effectively reversed DON-induced cytotoxicity via downregulating the expression of TNF-α, IL-6, and IL-lβ. In addition, S. boulardii significantly alleviated DON-induced phosphorylation and mRNA expression of p38 and further increased the expression of apoptosis regulation genes Bcl-xl and Bcl-2 and inhibited the activation of Bax. Our results suggest that S. boulardii could suppress DON-induced p38 MAPK pathway activation and reduce the expression of downstream inflammatory cytokines, as well as promote the expression of anti-apoptotic genes to inhibit apoptosis induced by DON in PAMC.
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Vanhoutte, Ilse; De Mets, Laura; De Boevre, Marthe; Uka, Valdet; Di Mavungu, José Diana; De Saeger, Sarah; De Gelder, Leen; Audenaert, Kris
2017-01-01
Mycotoxins are toxic metabolites produced by fungi. To mitigate mycotoxins in food or feed, biotransformation is an emerging technology in which microorganisms degrade toxins into non-toxic metabolites. To monitor deoxynivalenol (DON) biotransformation, analytical tools such as ELISA and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) are typically used. However, these techniques do not give a decisive answer about the remaining toxicity of possible biotransformation products. Hence, a bioassay using Lemna minor L. was developed. A dose–response analysis revealed significant inhibition in the growth of L. minor exposed to DON concentrations of 0.25 mg/L and higher. Concentrations above 1 mg/L were lethal for the plant. This bioassay is far more sensitive than previously described systems. The bioassay was implemented to screen microbial enrichment cultures, originating from rumen fluid, soil, digestate and activated sludge, on their biotransformation and detoxification capability of DON. The enrichment cultures originating from soil and activated sludge were capable of detoxifying and degrading 5 and 50 mg/L DON. In addition, the metabolites 3-epi-DON and the epimer of de-epoxy-DON (3-epi-DOM-1) were found as biotransformation products of both consortia. Our work provides a new valuable tool to screen microbial cultures for their detoxification capacity. PMID:28208799
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Vanhoutte, Ilse; De Mets, Laura; De Boevre, Marthe; Uka, Valdet; Di Mavungu, José Diana; De Saeger, Sarah; De Gelder, Leen; Audenaert, Kris
2017-02-13
Mycotoxins are toxic metabolites produced by fungi. To mitigate mycotoxins in food or feed, biotransformation is an emerging technology in which microorganisms degrade toxins into non-toxic metabolites. To monitor deoxynivalenol (DON) biotransformation, analytical tools such as ELISA and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) are typically used. However, these techniques do not give a decisive answer about the remaining toxicity of possible biotransformation products. Hence, a bioassay using Lemna minor L. was developed. A dose-response analysis revealed significant inhibition in the growth of L. minor exposed to DON concentrations of 0.25 mg/L and higher. Concentrations above 1 mg/L were lethal for the plant. This bioassay is far more sensitive than previously described systems. The bioassay was implemented to screen microbial enrichment cultures, originating from rumen fluid, soil, digestate and activated sludge, on their biotransformation and detoxification capability of DON. The enrichment cultures originating from soil and activated sludge were capable of detoxifying and degrading 5 and 50 mg/L DON. In addition, the metabolites 3-epi-DON and the epimer of de-epoxy-DON (3-epi-DOM-1) were found as biotransformation products of both consortia. Our work provides a new valuable tool to screen microbial cultures for their detoxification capacity.
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Devreese, Mathias; Broekaert, Nathan; Verbrugghe, Elin; De Saeger, Sarah; Audenaert, Kris; Haesebrouck, Freddy; Pasmans, Frank; Ducatelle, Richard; Croubels, Siska; Martel, An
2016-01-01
Seed-based pigeon diets could be expected to result in exposure of pigeons to mycotoxins such as deoxynivalenol (DON). Ingestion of low to moderate contamination levels of DON may impair intestinal health, immune function and/or pathogen fitness, resulting in altered host-pathogen interactions and thus different outcome of infections. Here we demonstrate that DON was one of the most frequently detected mycotoxins in seed-based racing pigeons feed, contaminating 5 out of 10 samples (range 177–1,466 μg/kg). Subsequently, a toxicokinetic analysis revealed a low absolute oral bioavailability (F) of DON in pigeons (30.4%), which is comparable to other avian species. Furthermore, semi-quantitative analysis using high-resolution mass spectrometry revealed that DON-3α-sulphate is the major metabolite of DON in pigeons after intravenous as well as oral administration. Following ingestion of DON contaminated feed, the intestinal epithelial cells are exposed to significant DON concentrations which eventually may affect intestinal translocation and colonization of bacteria. Feeding pigeons a DON contaminated diet resulted in an increased percentage of pigeons shedding Salmonella compared to birds fed control diet, 87 ± 17% versus 74 ± 13%, respectively. However, no impact of DON was observed on the Salmonella induced disease signs, organ lesions, faecal and organ Salmonella counts. The presented risk assessment indicates that pigeons are frequently exposed to mycotoxins such as DON, which can affect the outcome of a Salmonella infection. The increasing number of pigeons shedding Salmonella suggests that DON can promote the spread of the bacterium within pigeon populations. PMID:27997572
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Occurrence of deoxynivalenol in wheat flour, instant noodle and biscuits commercialised in Brazil.
Almeida, Adriana Palma de; Lamardo, Leda Conceição Antonia; Shundo, Luzia; Silva, Simone Alves da; Navas, Sandra Aparecida; Alaburda, Janete; Ruvieri, Valter; Sabino, Myrna
2016-12-01
A total of 134 samples, consisting of 58 wheat flour, 40 instant noodle and 36 biscuits, were analysed for the presence of deoxynivalenol (DON). The samples were obtained from retail markets of the city of São Paulo during the period 2010-2014. DON was determined by high performance liquid chromatography with ultraviolet detection and immunoaffinity sample clean-up. Method validation followed international guidelines. The LOD and LOQ were 60 and 200 µg kg -1 , respectively, considering the three different types of samples analysed. The lowest recovery found in this study was 91.8% with RSD 4.5% for instant noodles. DON was detected in 91.4%, 97.5% and 97.2% of samples wheat flour, instant noodles and biscuits, respectively, resulting in a total of 94.8% with levels ranging from LOD to 1720.0 µg kg -1 .
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Multi-mycotoxin contamination of couscous semolina commercialized in Morocco.
Zinedine, Abdellah; Fernández-Franzón, Mónica; Mañes, Jordi; Manyes, Lara
2017-01-01
The multi-mycotoxin contamination of ninety-eight (98) couscous semolina samples collected from various areas in Morocco was investigated in this study. Samples were surveyed for the presence of 22 mycotoxins (four aflatoxins, ochratoxin A, diacetoxiscyrpenol (DAS), three fumonisins, beauvericin (BEA), deoxynivalenol (DON), 15-acetyl-deoxynivalenol (15-ADON), 3-acetyl-deoxynivalenol (3-ADON), nivalenol (NIV), sterigmatocystin (STG), zearalenone (ZEA), four enniatins, T-2 and HT-2 toxins). Results showed that 96 out of 98 total couscous samples (98%) were contaminated by at least one mycotoxin. Enniatin B (ENB), Enniatin B1 (ENB1), Enniatin A1 (ENA1) and zearalenone (ZEA) have shown the highest incidences in contaminated samples. The dietary exposure was estimated to be 1.02, 0.57, 0.06, 0.57 and 0.3μg/kgbw/day for the sum of (DON+3-ADON+15-ADON), fumonisins (FB1+FB2+FB3), the sum of (T2+HT-2), NIV and ZEA, respectively. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Paulick, Marleen; Winkler, Janine; Kersten, Susanne; Schatzmayr, Dian; Frahm, Jana; Kluess, Jeannette; Schwartz-Zimmermann, Heidi Elisabeth; Dänicke, Sven
2018-02-01
The objective of the present study was to demonstrate the efficiency of the decontamination process applied to deoxynivalenol (DON)-contaminated maize by sodium sulphite (Na 2 SO 3 ) treatment in vivo. Additionally, in vitro characterisation of the toxicity of the DON sulphonates (DONS 1, 2 and 3 denote structurally different forms), the resulting DON metabolites, on peripheral blood mononuclear cells (PBMC) should substantiate the inactivation of DON. In a piglet experiment, both DON-contaminated maize and -uncontaminated control maize either untreated (DON-, CON-) or Na 2 SO 3 -treated (DON+, CON+) were mixed into feed and fed for 42 d starting from weaning. The results showed that feed intake and daily weight gain of animals fed DON- were significantly lower compared to animals fed CON- and CON+, whereas group DON+ reached the control level or even exceeded it. The feed-to-gain ratio was unaffected (p = 0.45). Furthermore, DON concentrations in plasma markedly reflected the diets' DON concentrations. These were < 0.1, < 0.1, 5.4 and 0.8 mg/kg feed for CON-, CON+, DON- and DON+, and amounted to 0.3, 0.4, 33.0 and 9.3 ng/ml in plasma, respectively. Whereas DONS 2 and 3 were detected in the DON+ diet, only DONS 2 was recovered in plasma. Regarding the toxicity of DONS, no or much lower toxicity was found compared to DON. DONS 1 and Na 2 SO 3 did not affect the viability of PBMC. At 32.71μM DONS2 the viability was reduced by 50% and thus this compound was less toxic than DON by a factor of 73. Consequently, wet preservation of maize with Na 2 SO 3 was an effective tool to avoid the adverse effects of DON on performance of piglets.
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Deoxynivalenol: rationale for development and application of a urinary biomarker.
Turner, Paul C; Burley, Victoria J; Rothwell, Joseph A; White, Kay L M; Cade, Janet E; Wild, Christopher P
2008-07-01
Mycotoxins are common dietary contaminants in most regions of the world. The frequency of exposure to the various families of mycotoxins is often dependent on geographic location, national wealth and related agricultural and regulatory infrastructure, combined with diversity of diet and degree of food sufficiency. Deoxynivalenol (DON) is a Fusarium mycotoxin that frequently contaminates wheat, corn and barley in temperate regions. A number of acute poisoning incidences have been linked to DON-contaminated foods and chronic exposure to lower levels of DON has been predicted in many regions. DON is a potent animal toxin and exposure in humans may cause gastroenteritis, growth faltering and immune toxicity. An ability to conduct accurate exposure assessment at the individual level is required to fully understand the potential health consequences for humans. To date, such exposure biomarkers have been lacking for many important mycotoxins, including DON. To better assess exposure to DON at the individual level, we have developed a robust urinary assay, incorporating immunoaffinity column (IAC) enrichment and LC-MS detection. Further refinement of this urinary assay, by inclusion of (13)C-DON as an internal standard, was then undertaken and tested within the UK. DON was frequently observed in urine and was significantly associated with cereal intake. A dietary intervention study demonstrated that avoiding wheat in the diet markedly reduced urinary levels of DON. This biomarker requires further validation but our initial data suggest it may provide a useful tool in epidemiological investigations of the potential health consequences of this common environmental toxin.
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Abdel-Wahhab, Mosaad A; El-Nekeety, Aziza A; Salman, Asmaa S; Abdel-Aziem, Sekena H; Mehaya, Fathy M; Hassan, Nabila S
2018-02-01
Deoxynivalenol (DON) is a Fusarium mycotoxin that frequently contaminates cereal and cereal-based food and induces liver injury. This study evaluated the protective role of silymarin nanoparticles (SILNPs) and inulin nanoparticles (INNPs) against DON-induced liver injury in rats. Eleven groups of rats were treated orally for 3 weeks as follows: the control group, DON-treated group (5 mg/kg b.w.); INNPs-treated groups at low (LD) or high (HD) dose (100 or 200 mg/kg b.w.); SILPNs-treated group (50 mg/kg b.w.); SILNPs plus INNPs(LD) or INNPs(HD)-treated groups; INNPs(LD) or INNPs(HD) plus DON-treated groups and DON plus SILNPs and INNPs(LD) or INNPs(HD)-treated groups. Blood and tissue samples were collected for different analyses. The results revealed that the practical sizes were 200 and 98 nm for SILNPs and INNPs respectively. DON increased liver enzymes activity, lipid profile, serum cytokines, number and percentage of chromosomal aberration, DNA fragmentation and comet score. It disturbed the oxidative stress markers, down regulated gene expression and induced histological changes in the liver tissue. Treatment with DON and SILNPs and/or INNPs at the two tested doses improved all the tested parameters and SILNPs plus INNPs(HD) normalized most of these parameters in DON-treated animals. SILNPs and INNPs could be promising candidates as hepatoprotective against DON or other hepatotoxins. Copyright © 2017 Elsevier Ltd. All rights reserved.
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Van De Walle, Jacqueline; Romier, Béatrice; Larondelle, Yvan; Schneider, Yves-Jacques
2008-04-01
Deoxynivalenol (DON) is the most prevalent trichothecene mycotoxin in crops in Europe and North America. In human intestinal Caco-2 cells, DON activates the mitogen-activated protein kinases (MAPKs). We hypothesized a link between DON ingestion and intestinal inflammation, and used Caco-2 cells to assess the effects of DON, at plausible intestinal concentrations (250-10,000 ng/ml), on inflammatory mediators acting downstream the MAPKs cascade i.e. activation of nuclear factor-kappaB (NF-kappaB) and interleukin-8 (IL-8) secretion. In addition, Caco-2 cells were co-exposed to pro-inflammatory stimuli in order to mimic an inflamed intestinal epithelium. Dose-dependent increases in NF-kappaB activity and IL-8 secretion were observed, reaching 1.4- and 7.6-fold, respectively using DON at 10 microg/ml. Phosphorylation of inhibitor-kappaB (IkappaB) increased (1.6-fold) at DON levels <0.5 microg/ml. Exposure of Caco-2 cells to pro-inflammatory agents, i.e. 25 ng/ml interleukin-1beta, 100 ng/ml tumor necrosis factor-alpha or 10 microg/ml lipopolysaccharides, activated NF-kappaB and increased IL-8 secretion. Synergistic interactions between these stimuli and DON were observed. These data show that DON induces NF-kappaB activation and IL-8 secretion dose-dependently in Caco-2 cells, and this effect was accentuated upon pro-inflammatory stimulation, suggesting DON exposure could cause or exacerbate intestinal inflammation.
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Rapid Analysis of Deoxynivalenol in Durum Wheat by FT-NIR Spectroscopy
De Girolamo, Annalisa; Cervellieri, Salvatore; Visconti, Angelo; Pascale, Michelangelo
2014-01-01
Fourier-transform-near infrared (FT-NIR) spectroscopy has been used to develop quantitative and classification models for the prediction of deoxynivalenol (DON) levels in durum wheat samples. Partial least-squares (PLS) regression analysis was used to determine DON in wheat samples in the range of <50–16,000 µg/kg DON. The model displayed a large root mean square error of prediction value (1,977 µg/kg) as compared to the EU maximum limit for DON in unprocessed durum wheat (i.e., 1,750 µg/kg), thus making the PLS approach unsuitable for quantitative prediction of DON in durum wheat. Linear discriminant analysis (LDA) was successfully used to differentiate wheat samples based on their DON content. A first approach used LDA to group wheat samples into three classes: A (DON ≤ 1,000 µg/kg), B (1,000 < DON ≤ 2,500 µg/kg), and C (DON > 2,500 µg/kg) (LDA I). A second approach was used to discriminate highly contaminated wheat samples based on three different cut-off limits, namely 1,000 (LDA II), 1,200 (LDA III) and 1,400 µg/kg DON (LDA IV). The overall classification and false compliant rates for the three models were 75%–90% and 3%–7%, respectively, with model LDA IV using a cut-off of 1,400 µg/kg fulfilling the requirement of the European official guidelines for screening methods. These findings confirmed the suitability of FT-NIR to screen a large number of wheat samples for DON contamination and to verify the compliance with EU regulation. PMID:25384107
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Rapid analysis of deoxynivalenol in durum wheat by FT-NIR spectroscopy.
De Girolamo, Annalisa; Cervellieri, Salvatore; Visconti, Angelo; Pascale, Michelangelo
2014-11-06
Fourier-transform-near infrared (FT-NIR) spectroscopy has been used to develop quantitative and classification models for the prediction of deoxynivalenol (DON) levels in durum wheat samples. Partial least-squares (PLS) regression analysis was used to determine DON in wheat samples in the range of <50-16,000 µg/kg DON. The model displayed a large root mean square error of prediction value (1,977 µg/kg) as compared to the EU maximum limit for DON in unprocessed durum wheat (i.e., 1,750 µg/kg), thus making the PLS approach unsuitable for quantitative prediction of DON in durum wheat. Linear discriminant analysis (LDA) was successfully used to differentiate wheat samples based on their DON content. A first approach used LDA to group wheat samples into three classes: A (DON ≤ 1,000 µg/kg), B (1,000 < DON ≤ 2,500 µg/kg), and C (DON > 2,500 µg/kg) (LDA I). A second approach was used to discriminate highly contaminated wheat samples based on three different cut-off limits, namely 1,000 (LDA II), 1,200 (LDA III) and 1,400 µg/kg DON (LDA IV). The overall classification and false compliant rates for the three models were 75%-90% and 3%-7%, respectively, with model LDA IV using a cut-off of 1,400 µg/kg fulfilling the requirement of the European official guidelines for screening methods. These findings confirmed the suitability of FT-NIR to screen a large number of wheat samples for DON contamination and to verify the compliance with EU regulation.
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Zhu, Yan; Hassan, Yousef I; Shao, Suqin; Zhou, Ting
2018-06-29
Deoxynivalenol (DON) is a type B trichothecene mycotoxin that is commonly detected in grains infested with Fusarium species. The maximum tolerated levels of DON in the majority of world's countries are restricted to 0.75 mg kg -1 within the human food chain and to less than 1-5 mg kg -1 in animal feed depending on the feed material and/or animal species due to DON's short and long-term adverse effects on human health and animal productivity. The ability to accurately analyze DON and some of its fungal/bacterial metabolites is increasingly gaining a paramount importance in food/feed analysis and research. In this study, we used the immuno-affinity approach to enrich and detect DON and three of its bacterial metabolites, namely 3-epi-DON, 3-keto-DON, and deepoxy-DON (DOM-1). The optimized enrichment step coupled with high performance liquid chromatography can accurately and reproducibly quantify the aforementioned metabolites in feed matrixes (silage extract as an example in this case). It minimizes any background interface and provides a fast and easy-to-operate protocol for the analytical determination of such metabolites. More importantly, the presented data demonstrates the ability of the utilized monoclonal antibody, generated originally to capture DON in Enzyme-Linked Immunosorbent Assays (ELISA), to cross react with three less/non-toxic DON metabolites. This raises the concerns about the genuine need to account for such cross-reactivity when DON contamination is assessed through an immuno-affinity based analyses using the investigated antibody. Crown Copyright © 2018. Published by Elsevier B.V. All rights reserved.
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Occurrence of deoxynivalenol and zearalenone in brewing barley grains from Brazil.
Piacentini, Karim C; Rocha, L O; Savi, G D; Carnielli-Queiroz, L; Almeida, F G; Minella, E; Corrêa, B
2018-03-09
Barley (Hordeum vulgare L.) is an important cereal crop for food and represents one of the main ingredients in beer production. Considering the importance of barley and its derived products, the knowledge about the mycotoxin contamination in the barley production is essential in order to assess its safety. In this study, the levels of deoxynivalenol (DON) and zearalenone (ZEN) in brewing barley were determined using a LC-MS/MS method. A survey was conducted in 2015 to estimate the mycotoxin levels in these products (n = 76) from four crop regions in Brazil. The results showed high levels of DON and ZEN in the analyzed samples, with contamination levels of 94 and 73.6%, respectively. The mean levels of DON and ZEN ranged from 1700 to 7500 μg/kg and from 300 to 630 μg/kg, respectively. Barley samples from regions 1 and 2 presented higher levels of ZEN and DON, respectively, and those from region 4 presented lower levels of both. Co-occurrence of DON and ZEN was seen in the majority of the barley grain samples, and the mycotoxin content was above the maximum levels established by the Brazilian and European regulations.
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Wu, Li; Wang, Bujun
2016-07-01
We hereby report the transformation of deoxynivalenol (DON) and its acetylated derivatives (3-ADON and 15-ADON) by spiking targeted mycotoxins to Fusarium mycotoxin-free flour in the process of making Chinese steamed bread (CSB). The impacts of pH, yeast level, and steaming time on the transformation of 3-ADON to DON were investigated. DON, 3-ADON, and 15-ADON were analyzed by UPLC-MS/MS. Spiked DON was stable throughout the CSB making process. Spiked 3-ADON and 15-ADON were partially deacetylated and transformed to DON during kneading (54.1-60.0% and 59.3-77.5%, respectively), fermentation (64.0-76.9% and 78.2-91.6%, respectively), and steaming (47.2-52.7% and 52.4-61.9%, respectively). The ADONs level increased after steaming compared with their level in the previous step. The pH level and steaming duration significantly (P<0.05) affected the conversion of 3-ADON during the CSB making process. Briefly, alkaline conditions and short steaming times favored the deacetylation of 3-ADON. The level of yeast did not remarkably (P<0.05) alter the transformation between ADONs and DON. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Pralatnet, Sasithorn; Poapolathep, Saranya; Giorgi, Mario; Imsilp, Kanjana; Kumagai, Susumu; Poapolathep, Amnart
2016-07-01
One hundred wheat product samples (50 instant noodle samples and 50 bread samples) were collected from supermarkets in Bangkok, Thailand. Deoxynivalenol (DON) and aflatoxin B1 (AFB1) contamination in these products was analyzed using a validated liquid chromatography-tandem mass spectrometry method. The limit of quantification values of DON and AFB1 in the instant noodles and bread were 2 and 1 ng g(-1), respectively. The survey found that DON was quantifiable in 40% of collected samples, in 2% of noodles (0.089 μg g(-1)), and in 78% of breads (0.004 to 0.331 μg g(-1)). AFB1 was below the limit of quantification of the method in all of the tested samples. The results suggest that the risk of DON exposure via noodles and breads is very low in urban areas of Thailand. No risk can be attributable to AFB1 exposure in the same food matrices, but further studies with a larger sample size are needed to confirm these data.
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Sanders, Melanie; McPartlin, Daniel; Moran, Kara; Guo, Yirong; Eeckhout, Mia; O’Kennedy, Richard; De Saeger, Sarah; Maragos, Chris
2016-01-01
A sample preparation method was developed for the screening of deoxynivalenol (DON) in wheat and wheat dust. Extraction was carried out with water and was successful due to the polar character of DON. For detection, an enzyme-linked immunosorbent assay (ELISA) was compared to the sensor-based techniques of surface plasmon resonance (SPR) and biolayer interferometry (BLI) in terms of sensitivity, affinity and matrix effect. The matrix effects from wheat and wheat dust using SPR were too high to further use this screenings method. The preferred ELISA and BLI methods were validated according to the criteria established in Commission Regulation 519/2014/EC and Commission Decision 2002/657/EC. A small survey was executed on 16 wheat lots and their corresponding dust samples using the validated ELISA method. A linear correlation (r = 0.889) was found for the DON concentration in dust versus the DON concentration in wheat (LOD wheat: 233 μg/kg, LOD wheat dust: 458 μg/kg). PMID:27077883
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Han, Jun; Wang, Qiao-Chu; Zhu, Cheng-Cheng
Deoxynivalenol (DON) is a widespread trichothecene mycotoxin which contaminates agricultural staples and elicits a complex spectrum of toxic effects on humans and animals. It has been shown that DON impairs oocyte maturation, reproductive function and causes abnormal fetal development in mammals; however, the mechanisms remain unclear. In the present study, we investigate the possible reasons of the toxic effects of DON on porcine oocytes. Our results showed that DON significantly inhibited porcine oocyte maturation and disrupted meiotic spindle by reducing p-MAPK protein level, which caused retardation of cell cycle progression. In addition, up-regulated LC3 protein expression and aberrant Lamp2, LC3more » and mTOR mRNA levels were observed with DON exposure, together with Annexin V-FITC staining assay analysis, these results indicated that DON treatment induced autophagy/apoptosis in porcine oocytes. We also showed that DON exposure increased DNA methylation level in porcine oocytes through altering DNMT3A mRNA levels. Histone methylation levels were also changed showing with increased H3K27me3 and H3K4me2 protein levels, and mRNA levels of their relative methyltransferase genes, indicating that epigenetic modifications were affected. Taken together, our results suggested that DON exposure reduced porcine oocytes maturation capability through affecting cytoskeletal dynamics, cell cycle, autophagy/apoptosis and epigenetic modifications. - Highlights: • DON exposure disrupted meiotic spindle by reducing p-MAPK expression. • DON exposure caused retardation of cell cycle progression in porcine oocytes. • DON triggered autophagy and early-apoptosis in porcine oocytes. • DON exposure led to aberrant epigenetic modifications in porcine oocytes.« less
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Frequent biphasic cellular responses of permanent fish cell cultures to deoxynivalenol (DON)
DOE Office of Scientific and Technical Information (OSTI.GOV)
Pietsch, Constanze, E-mail: constanze.pietsch@unibas.ch; Bucheli, Thomas D.; Wettstein, Felix E.
Contamination of animal feed with mycotoxins is a major problem for fish feed mainly due to usage of contaminated ingredients for production and inappropriate storage of feed. The use of cereals for fish food production further increases the risk of a potential contamination. Potential contaminants include the mycotoxin deoxynivalenol (DON) which is synthesized by globally distributed fungi of the genus Fusarium. The toxicity of DON is well recognized in mammals. In this study, we confirm cytotoxic effects of DON in established permanent fish cell lines. We demonstrate that DON is capable of influencing the metabolic activity and cell viability inmore » fish cells as determined by different assays to indicate possible cellular targets of this toxin. Evaluation of cell viability by measurement of membrane integrity, mitochondrial activity and lysosomal function after 24 h of exposure of fish cell lines to DON at a concentration range of 0-3000 ng ml{sup -1} shows a biphasic effect on cells although differences in sensitivity occur. The cell lines derived from rainbow trout are particularly sensitive to DON. The focus of this study lies, furthermore, on the effects of DON at different concentrations on production of reactive oxygen species (ROS) in the different fish cell lines. The results show that DON mainly reduces ROS production in all cell lines that were used. Thus, our comparative investigations reveal that the fish cell lines show distinct species-related endpoint sensitivities that also depend on the type of tissue from which the cells were derived and the severity of exposure. - Highlights: > DON uptake by cells is not extensive. > All fish cell lines are sensitive to DON. > DON is most cytotoxic to rainbow trout cells. > Biphasic cellular responses were frequently observed. > Our results are similar to studies on mammalian cell lines.« less
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Poppenberger, B; Berthiller, F; Lucyshyn, D; Schuhmacher, R; Krska, R; Adam, G
2005-06-01
First results of the GEN-AU pilot project "Fusarium virulence and plant resistance mechanisms" are reported. Employing genetically engineered yeast strains we have been able to clone genes from the model plantArabidopsis thaliana encoding UDP-glucosyltransferases which can inactivate deoxynivalenol (DON) and zearalenone (ZON). The structure of the metabolites produced by the transformed yeast strains were determined by LC-MS/MS as DON-3O-glucoside and ZON-4O-glucoside, respectively. ZON and derivatives added to glucosyltransferase expressing yeast cultures are converted into the corresponding glucosides in very high yield, opening an efficient way to produce reference materials for these masked mycotoxins.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Van De Walle, Jacqueline; Sergent, Therese; Piront, Neil
Deoxynivalenol (DON), one of the most common mycotoxin contaminants of raw and processed cereal food, adversely affects the gastrointestinal tract. Since DON acts as a protein synthesis inhibitor, the constantly renewing intestinal epithelium could be particularly sensitive to DON. We analyzed the toxicological effects of DON on intestinal epithelial protein synthesis and barrier integrity. Differentiated Caco-2 cells, as a widely used model of the human intestinal barrier, were exposed to realistic intestinal concentrations of DON (50, 500 and 5000 ng/ml) during 24 h. DON caused a concentration-dependent decrease in total protein content associated with a reduction in the incorporation ofmore » [{sup 3}H]-leucine, demonstrating its inhibitory effect on protein synthesis. DON simultaneously increased the paracellular permeability of the monolayer as reflected through a decreased transepithelial electrical resistance associated with an increased paracellular flux of the tracer [{sup 3}H]-mannitol. A concentration-dependent reduction in the expression level of the tight junction constituent claudin-4 was demonstrated by Western blot, which was not due to diminished transcription, increased degradation, or NF-{kappa}B, ERK or JNK activation, and was also observed for a tight junction independent protein, i.e. intestinal alkaline phosphatase. These results demonstrate a dual toxicological effect of DON on differentiated Caco-2 cells consisting in an inhibition of protein synthesis as well as an increase in monolayer permeability, and moreover suggest a possible link between them through diminished synthesis of the tight junction constituent claudin-4.« less
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Ji, Jian; Zhu, Pei; Cui, Fangchao; Pi, Fuwei; Zhang, Yinzhi; Li, Yun; Wang, Jiasheng; Sun, Xiulan
2017-01-01
Metabolic profiling in liver and serum of mice was studied for the combined toxic effects of deoxynivalenol (DON) and zearalenone (ZEN), through gas chromatography mass spectrum. The spectrum of serum and liver sample of mice, treated with individual 2 mg/kg DON, 20 mg/kg ZEN, and the combined DON + ZEN with final concentration 2 mg/kg DON and 20 mg/kg ZEN for 21 days, were deconvoluted, aligned and identified with MS DIAL. The data matrix was processed with univariate analysis and multivariate analysis for selection of metabolites with variable importance for the projection (VIP) > 1, t-test p value < 0.05. The metabolic pathway analysis was performed with MetaMapp and drawn by CytoScape. Results show that the combined DON and ZEN treatment has an obvious “antagonistic effect” in serum and liver tissue metabolic profiling of mice. The blood biochemical indexes, like alkaline phosphatase, alanine transaminase, and albumin (ALB)/globulin (GLO), reveal a moderated trend in the combined DON + ZEN treatment group, which is consistent with histopathological examination. The metabolic pathway analysis demonstrated that the combined DON and ZEN treatment could down-regulate the valine, leucine and isoleucine biosynthesis, glycine, serine and threonine metabolism, and O-glycosyl compounds related glucose metabolism in liver tissue. The metabolic profiling in serum confirmed the finding that the combined DON and ZEN treatment has an “antagonistic effect” on liver metabolism of mice. PMID:28075412
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Kobayashi-Hattori, Kazuo; Amuzie, Chidozie J; Flannery, Brenna M; Pestka, James J
2011-07-01
To characterize the effects of ingesting the common foodborne mycotoxin deoxynivalenol (DON) on body weight and composition in the high-fat (HF) diet-induced obese mice, a model of human obesity. Female B6C3F1 mice were initially fed HF diets containing 45% kcal (HF45) or 60% kcal (HF60) as fat for 94 days to induce obesity. Half of each group was either continued on unamended HF diets or fed HF diets containing 10 mg/kg DON (DON-HF45 or DON-HF60) for another 54 days. Additional control mice were fed a low-fat (LF) diet containing 10% kcal as fat for the entire 148-day period. DON induced rapid decreases in body weights and fat mass, which stabilized to those of the LF control within 11 days. These effects corresponded closely to a robust transient decrease in food consumption. While lean body mass did not decline in DON-fed groups, further increases were suppressed. DON exposure reduced plasma insulin, leptin, insulin-like growth factor 1, and insulin-like growth factor acid labile subunit as well as increased hypothalamic mRNA level of the orexigenic agouti-related protein. DON-mediated effects on body weight, fat mass, food intake, and hormonal levels in obese mice were consistent with a state of chronic energy restriction. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Antonissen, Gunther; Van Immerseel, Filip; Pasmans, Frank; Ducatelle, Richard; Haesebrouck, Freddy; Timbermont, Leen; Verlinden, Marc; Janssens, Geert Paul Jules; Eeckhaut, Venessa; Eeckhout, Mia; De Saeger, Sarah; Hessenberger, Sabine; Martel, An; Croubels, Siska
2014-01-01
Both mycotoxin contamination of feed and Clostridium perfringens-induced necrotic enteritis have an increasing global economic impact on poultry production. Especially the Fusarium mycotoxin deoxynivalenol (DON) is a common feed contaminant. This study aimed at examining the predisposing effect of DON on the development of necrotic enteritis in broiler chickens. An experimental Clostridium perfringens infection study revealed that DON, at a contamination level of 3,000 to 4,000 µg/kg feed, increased the percentage of birds with subclinical necrotic enteritis from 20±2.6% to 47±3.0% (P<0.001). DON significantly reduced the transepithelial electrical resistance in duodenal segments (P<0.001) and decreased duodenal villus height (P = 0.014) indicating intestinal barrier disruption and intestinal epithelial damage, respectively. This may lead to an increased permeability of the intestinal epithelium and decreased absorption of dietary proteins. Protein analysis of duodenal content indeed showed that DON contamination resulted in a significant increase in total protein concentration (P = 0.023). Furthermore, DON had no effect on in vitro growth, alpha toxin production and netB toxin transcription of Clostridium perfringens. In conclusion, feed contamination with DON at concentrations below the European maximum guidance level of 5,000 µg/kg feed, is a predisposing factor for the development of necrotic enteritis in broilers. These results are associated with a negative effect of DON on the intestinal barrier function and increased intestinal protein availability, which may stimulate growth and toxin production of Clostridium perfringens.
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Effects of deoxynivalenol on content of chloroplast pigments in barley leaf tissues.
Bushnell, W R; Perkins-Veazie, P; Russo, V M; Collins, J; Seeland, T M
2010-01-01
To understand further the role of deoxynivalenol (DON) in development of Fusarium head blight (FHB), we investigated effects of the toxin on uninfected barley tissues. Leaf segments, 1 to 1.2 cm long, partially stripped of epidermis were floated with exposed mesophyll in contact with DON solutions. In initial experiments with the leaf segments incubated in light, DON at 30 to 90 ppm turned portions of stripped tissues white after 48 to 96 h. The bleaching effect was greatly enhanced by addition of 1 to 10 mM Ca(2+), so that DON at 10 to 30 ppm turned virtually all stripped tissues white within 48 h. Content of chlorophylls a and b and of total carotenoid pigment was reduced. Loss of electrolytes and uptake of Evans blue indicated that DON had a toxic effect, damaging plasmalemmas in treated tissues before chloroplasts began to lose pigment. When incubated in the dark, leaf segments also lost electrolytes, indicating DON was toxic although the tissues remained green. Thus, loss of chlorophyll in light was due to photobleaching and was a secondary effect of DON, not required for toxicity. In contrast to bleaching effects, some DON treatments that were not toxic kept tissues green without bleaching or other signs of injury, indicating senescence was delayed compared with slow yellowing of untreated leaf segments. Cycloheximide, which like DON, inhibits protein synthesis, also bleached some tissues and delayed senescence of others. Thus, the effects of DON probably relate to its ability to inhibit protein synthesis. With respect to FHB, the results suggest DON may have multiple roles in host cells of infected head tissues, including delayed senescence in early stages of infection and contributing to bleaching and death of cells in later stages.
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Ghareeb, K; Awad, W A; Böhm, J
2012-04-01
Although acute mycotoxicoses are rare in poultry production, chronic exposure to low levels of mycotoxins is responsible for reduced productivity and increased susceptibility to infectious diseases. Deoxynivalenol (DON) is known to modulate immune function, but only a few studies have investigated the effect of DON on the vaccinal immune response. In addition, the effects of Mycofix select (Biomin GmbH, Herzogenburg, Austria) supplementation to DON-contaminated broiler diets have not yet been demonstrated. Therefore, an experiment with 1-d-old male broilers (Ross 308) was carried out to examine the effects of feeding DON-contaminated low-protein grower diets on performance, serum biochemical parameters, lymphoid organ weight, and antibody titers to infectious bronchitis vaccination in serum and to evaluate the effects of Mycofix select dietary supplementation in either the presence or absence of DON in broilers. In total, thirty-two 1-d-old broiler chicks were randomly assigned to 1 of the 4 dietary treatments for 5 wk. The dietary treatments were 1) control; 2) artificially contaminated diets with 10 mg of DON/kg of diet; 3) DON-contaminated diets supplemented with Mycofix select; and 4) control diet supplemented with Mycofix select. Feeding of contaminated diets decreased (P = 0.000) the feed intake, BW (P = 0.001), BW gain (P = 0.044), and feed efficiency during the grower phase. Deoxynivalenol affected the blood biochemistry, whereas plasma total protein and uric acid concentrations in birds fed contaminated grains were decreased compared with those of the controls. Moreover, in birds fed contaminated feeds, there was a tendency to reduce triglycerides in the plasma (P = 0.090), suggesting that DON in the diets affected protein and lipid metabolism in broiler chickens. The feeding of contaminated diets altered the immune response in broilers by reducing the total lymphocyte count. Similarly, the antibody response against infectious bronchitis vaccination antigens was decreased (P = 0.003) after feeding contaminated diets, compared with the controls. Moreover, contamination of the broiler diet with DON increased the heteropil:lymphocyte ratio (stress index), suggesting that DON elevated the physiological stress responses of broilers. However, feeding of DON-containing diets did not alter the other plasma constituents, including activities of enzymes. Mycofix select addition to the DON-contaminated feed led to normal immunological and physiological functions in broilers that were comparable with those of the control group, indicating that the addition of the additive to the DON-contaminated feed of the broilers effectively alleviated the alterations caused by DON. It was concluded that broiler performance and some blood and immunological parameters were adversely affected by feeding diets contaminated with the Fusarium mycotoxin DON. However, the dietary Mycofix select supplementation as a detoxifying agent was successful in overcoming the mycotoxin-related effects.
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Lindblad, M; Börjesson, T; Hietaniemi, V; Elen, O
2012-01-01
The relationship between weather data and agronomical factors and deoxynivalenol (DON) levels in oats was examined with the aim of developing a predictive model. Data were collected from a total of 674 fields during periods of up to 10 years in Finland, Norway and Sweden, and included DON levels in the harvested oats crop, agronomical factors and weather data. The results show that there was a large regional variation in DON levels, with higher levels in one region in Norway compared with other regions in Norway, Finland and Sweden. In this region the median DON level was 1000 ng g⁻¹ and the regulatory limit for human consumption (1750 ng g⁻¹) was exceeded in 28% of the samples. In other regions the median DON levels ranged from 75 to 270 ng g⁻¹, and DON levels exceeded 1750 ng g⁻¹ in 3-8% of the samples. Including more variables than region in a multiple regression model only increased the adjusted coefficient of determination from 0.17 to 0.24, indicating that very little of the variation in DON levels could be explained by weather data or agronomical factors. Thus, it was not possible to predict DON levels based on the variables included in this study. Further studies are needed to solve this problem. Apparently the infection and/or growth of DON producing Fusarium species are promoted in certain regions. One possibility may be to study the species distribution of fungal communities and their changes during the oats cultivation period in more detail.
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Therapeutic effects of glutamic acid in piglets challenged with deoxynivalenol.
Wu, Miaomiao; Xiao, Hao; Ren, Wenkai; Yin, Jie; Tan, Bie; Liu, Gang; Li, Lili; Nyachoti, Charles Martin; Xiong, Xia; Wu, Guoyao
2014-01-01
The mycotoxin deoxynivalenol (DON), one of the most common food contaminants, primarily targets the gastrointestinal tract to affect animal and human health. This study was conducted to examine the protective function of glutamic acid on intestinal injury and oxidative stress caused by DON in piglets. Twenty-eight piglets were assigned randomly into 4 dietary treatments (7 pigs/treatment): 1) uncontaminated control diet (NC), 2) NC+DON at 4 mg/kg (DON), 3) NC+2% glutamic acid (GLU), and 4) NC+2% glutamic acid + DON at 4 mg/kg (DG). At day 15, 30 and 37, blood samples were collected to determine serum concentrations of CAT (catalase), T-AOC (total antioxidant capacity), H2O2 (hydrogen peroxide), NO (nitric oxide), MDA (maleic dialdehyde), DAO (diamine oxidase) and D-lactate. Intestinal morphology, and the activation of Akt/mTOR/4EBP1 signal pathway, as well as the concentrations of H2O2, MDA, and DAO in kidney, liver and small intestine, were analyzed at day 37. Results showed that DON significantly (P<0.05) induced oxidative stress in piglets, while this stress was remarkably reduced with glutamic acid supplementation according to the change of oxidative parameters in blood and tissues. Meanwhile, DON caused obvious intestinal injury from microscopic observations and permeability indicators, which was alleviated by glutamic acid supplementation. Moreover, the inhibition of DON on Akt/mTOR/4EBP1 signal pathway was reduced by glutamic acid supplementation. Collectively, these data suggest that glutamic acid may be a useful nutritional regulator for DON-induced damage manifested as oxidative stress, intestinal injury and signaling inhibition.
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Therapeutic Effects of Glutamic Acid in Piglets Challenged with Deoxynivalenol
Ren, Wenkai; Yin, Jie; Tan, Bie; Liu, Gang; Li, Lili; Nyachoti, Charles Martin; Xiong, Xia; Wu, Guoyao
2014-01-01
The mycotoxin deoxynivalenol (DON), one of the most common food contaminants, primarily targets the gastrointestinal tract to affect animal and human health. This study was conducted to examine the protective function of glutamic acid on intestinal injury and oxidative stress caused by DON in piglets. Twenty-eight piglets were assigned randomly into 4 dietary treatments (7 pigs/treatment): 1) uncontaminated control diet (NC), 2) NC+DON at 4 mg/kg (DON), 3) NC+2% glutamic acid (GLU), and 4) NC+2% glutamic acid + DON at 4 mg/kg (DG). At day 15, 30 and 37, blood samples were collected to determine serum concentrations of CAT (catalase), T-AOC (total antioxidant capacity), H2O2 (hydrogen peroxide), NO (nitric oxide), MDA (maleic dialdehyde), DAO (diamine oxidase) and D-lactate. Intestinal morphology, and the activation of Akt/mTOR/4EBP1 signal pathway, as well as the concentrations of H2O2, MDA, and DAO in kidney, liver and small intestine, were analyzed at day 37. Results showed that DON significantly (P<0.05) induced oxidative stress in piglets, while this stress was remarkably reduced with glutamic acid supplementation according to the change of oxidative parameters in blood and tissues. Meanwhile, DON caused obvious intestinal injury from microscopic observations and permeability indicators, which was alleviated by glutamic acid supplementation. Moreover, the inhibition of DON on Akt/mTOR/4EBP1 signal pathway was reduced by glutamic acid supplementation. Collectively, these data suggest that glutamic acid may be a useful nutritional regulator for DON-induced damage manifested as oxidative stress, intestinal injury and signaling inhibition. PMID:24984001
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Co-occurrence and distribution of deoxynivalenol, nivalenol and zearalenone in wheat from Brazil.
Calori-Domingues, Maria Antonia; Bernardi, Carolina Maria Gil; Nardin, Mariana Sartori; de Souza, Gláucia Vendramini; Dos Santos, Fernanda Gregório Ribeiro; Stein, Mirella de Abreu; Gloria, Eduardo Micotti da; Dias, Carlos Tadeu Dos Santos; de Camargo, Adriano Costa
2016-06-01
Fusarium mycotoxins deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN) were investigated in wheat from the 2009 and 2010 crop years. Samples (n = 745) from commercial fields were collected in four wheat producing regions (WPR) which differed in weather conditions. Analyses were performed using HPLC-DAD. Contamination with ZEN, DON and NIV occurred in 56, 86 and 50%, respectively. Also, mean concentrations were different: DON = 1046 µg kg(-1), NIV < 100 µg kg(-1) and ZEN = 82 µg kg(-1). Co-occurrence of ZEN, DON and NIV was observed in 74% of the samples from 2009 and in 12% from 2010. Wet/cold region WPR I had the highest mycotoxin concentration. Wet/moderately hot region WPR II had the lowest mycotoxin levels. Furthermore, the mean concentration of each mycotoxin was higher in samples from 2009 as compared with those from 2010. Precipitation during flowering or harvest periods may explain these results.
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Piacentini, Karim C; Rocha, Liliana O; Fontes, Lívia C; Carnielli, Lorena; Reis, Tatiana A; Corrêa, Benedito
2017-03-01
Worldwide, barley is the main source of carbohydrate in the brewing process. However, corn is often used as an adjunct to improve and accelerate the fermentation process. Considering that, these two substrates are susceptible to fungal contamination as well as mycotoxins. The objective of the current study is to determine the incidence of the mycotoxins deoxynivalenol (DON) and fumonisin B 1 (FB 1 ) in industrial beers. The method applied for mycotoxin analyses included high performance liquid chromatography . The mean levels for recovery experiments were 89.6% for DON and 93.3% for FB 1 . DON was not detected in any of the analyzed samples whereas FB 1 was found in 49% of the 114 samples. The current survey demonstrated levels of FB 1 contamination in industrial beer, possibly due to the addition of contaminated adjuncts. It is necessary to establish maximum levels of mycotoxins in beer in Brazil and other countries in order to reduce health risks. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Deoxynivalenol and zearalenone in Fusarium-contaminated wheat in Mexico City.
González-Osnaya, L; Farrés, A
2011-01-01
Fusarium spp. invasion causes head blight, a destructive disease in the world's main wheat-growing areas, and deoxynivalenol (DON) and zearalenone (ZEA) contamination in cereal-based products. No data are available on the relationship between Fusarium spp. on commercial wheat samples in Mexico City and the presence of mycotoxins. A total of 30 wheat samples were subject to a PCR method involving genes of the trichothecene and zearalenone biosynthesis pathways to detect the presence of Fusarium. Detection and quantification of DON and ZEA was performed using liquid chromatography coupled to UV detection. PCR indicated the presence of the Tri5 and PKS4 genes in 16.7 and 23.3% of samples, respectively. DON and ZEA contamination was found in 51.2 and 71.4% of samples, respectively, where a positive amplification was obtained. This work presents up-to-date information on mycotoxin contamination in Mexico, where improved contamination/exposure data and firm control/monitoring measures are needed.
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Janaviciene, Sigita; Mankeviciene, Audrone; Suproniene, Skaidre; Kochiieru, Yuliia; Keriene, Ilona
2018-02-22
Deoxynivalenol (DON) together with two acetylated derivatives, 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON) occurs in cereal grains and their products. Co-occurrence of DON and acetylated derivatives in cereal grain is detected worldwide. Until now, DON and its derivatives have been considered equally toxic by health authorities. In this study, we analysed 103 samples of spring wheat grain, originating from the fields of different production systems in Lithuania, for the co-occurrence of type-B trichothecenes (DON, 3-ADON, 15-ADON). The samples were classified according to the production system-organic, sustainable and intensive. Mycotoxin levels in the spring wheat grain samples were determined by the HPLC method with UV detection. The type-B trichothecenes were found to be present at higher concentrations in the grain from the intensive production system. Eighty-one percent of the spring wheat grain samples from the intensive production system were co-contaminated with a combination of DON+3-ADON+15-ADON, 1% with DON+3-ADON. Additionally, DON+15-ADON and DON were found in 5% and 10% of the tested samples, respectively. Two percent of the samples were free from mycotoxins. In the grain samples from the sustainable production system, DON and a combination of DON+3-ADON showed a higher incidence - 47% and 23%, respectively. The samples with a combination of DON+3-ADON+15-ADON accounted for 18%. Completely different results were obtained from the analyses of organic grain samples. A large number of the organic spring wheat grain samples were contaminated with DON+3-ADON (55%) or DON (36%). The combination of DON+3-ADON+15-ADON was not present, while DON+15-ADON was present in 9% of the samples tested. The production systems did not lead to significant differences in mycotoxin levels, although a trend toward higher incidence and higher contamination was observed for the samples from the intensive and sustainable production systems.
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The Food Contaminant Mycotoxin Deoxynivalenol Inhibits the Swallowing Reflex in Anaesthetized Rats
Abysique, Anne; Tardivel, Catherine; Troadec, Jean-Denis; Félix, Bernadette
2015-01-01
Deoxynivalenol (DON), one of the most abundant mycotoxins found on cereals, is known to be implicated in acute and chronic illnesses in both humans and animals. Among the symptoms, anorexia, reduction of weight gain and decreased nutrition efficiency were described, but the mechanisms underlying these effects on feeding behavior are not yet totally understood. Swallowing is a major motor component of ingestive behavior which allows the propulsion of the alimentary bolus from the mouth to the esophagus. To better understand DON effects on ingestive behaviour, we have studied its effects on rhythmic swallowing in the rat, after intravenous and central administration. Repetitive electrical stimulation of the superior laryngeal nerve or of the tractus solitarius, induces rhythmic swallowing that can be recorded using electromyographic electrodes inserted in sublingual muscles. Here we provide the first demonstration that, after intravenous and central administration, DON strongly inhibits the swallowing reflex with a short latency and in a dose dependent manner. Moreover, using c-Fos staining, a strong neuronal activation was observed in the solitary tract nucleus which contains the central pattern generator of swallowing and in the area postrema after DON intravenous injection. Our data show that DON modifies swallowing and interferes with central neuronal networks dedicated to food intake regulation. PMID:26192767
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Adesso, Simona; Autore, Giuseppina; Quaroni, Andrea; Popolo, Ada; Severino, Lorella; Marzocco, Stefania
2017-12-11
Fusarium mycotoxins are fungal metabolites whose ability to affect cereal grains as multi-contaminants is progressively increasing. The trichothecene mycotoxins nivalenol (NIV) and deoxynivalenol (DON) are often found in almost all agricultural commodities worldwide. They are able to affect animal and human health, including at the intestinal level. In this study, NIV, both alone and in combination with DON, induced inflammation and increased the inflammatory response induced by lipopolysaccharide (LPS) plus Interferon-γ (IFN) in the non-tumorigenic intestinal epithelial cell line (IEC-6). The inflammatory response induced by NIV and DON involves tumor necrosis factor-α (TNF-α) production, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression, nitrotyrosine formation, reactive oxygen species (ROS) release, Nuclear Factor-κB (NF-κB), Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and inflammasome activation. The pro-inflammatory effect was strongly induced by NIV and by the mycotoxin mixture, when compared to DON alone. Mechanistic studies indicate a pivotal role for ROS in the observed pro-inflammatory effects induced by mycotoxins. In this study, the interactions between NIV and DON point out the importance of their food co-contamination, further highlighting the risk assessment process that is of growing concern.
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Yu, Miao; Chen, Liangkai; Peng, Zhao; Nüssler, Andreas K; Wu, Qinghua; Liu, Liegang; Yang, Wei
2017-06-01
Deoxynivalenol (DON) is a toxic fungal secondary metabolite produced by molds of the Fusarium genus, and it is known to cause a spectrum of diseases both in humans and animals, such as emesis, diarrhea, anorexia, immunotoxicity, hematological disorders, impairment of maternal reproduction, and fetal development. The recently revealed teratogenic potential of DON has received much attention. In various animal models, it has been shown that DON led to skeletal deformities of the fetus. However, the underlying mechanisms are not yet fully understood, and toxicological data are also scarce. Several animal research studies highlight the potential link between morphological abnormalities and changes of autophagy in the reproductive system. Because autophagy is involved in fetal development, maintenance of placental function, and bone remodeling, this mechanism has become a high priority for future research. The general aim of the present review is to deliver a comprehensive overview of the current state of knowledge of DON-induced reproductive toxicity in different animal models and to provide some prospective ideas for further research. The focus of the current review is to summarize toxic and negative effects of DON exposure on the reproductive system and the potential underlying molecular mechanisms in various animal models. Copyright © 2017 Elsevier B.V. All rights reserved.
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The mycotoxin deoxynivalenol facilitates allergic sensitization to whey in mice.
Bol-Schoenmakers, M; Braber, S; Akbari, P; de Graaff, P; van Roest, M; Kruijssen, L; Smit, J J; van Esch, B C A M; Jeurink, P V; Garssen, J; Fink-Gremmels, J; Pieters, R H H
2016-11-01
Intestinal epithelial stress or damage may contribute to allergic sensitization against certain food antigens. Hence, the present study investigated whether impairment of intestinal barrier integrity by the mycotoxin deoxynivalenol (DON) contributes to the development of whey-induced food allergy in a murine model. C3H/HeOuJ mice, orally exposed to DON plus whey once a week for 5 consecutive weeks, showed whey-specific IgG1 and IgE in serum and an acute allergic skin response upon intradermal whey challenge, although early initiating mechanisms of sensitization in the intestine appeared to be different compared with the widely used mucosal adjuvant cholera toxin (CT). Notably, DON exposure modulated tight-junction mRNA and protein levels, and caused an early increase in IL-33, whereas CT exposure affected intestinal γδ T cells. On the other hand, both DON- and CT-sensitized mice induced a time-dependent increase in the soluble IL-33 receptor ST2 (IL-1R1) in serum, and enhanced local innate lymphoid cells type 2 cell numbers. Together, these results demonstrate that DON facilitates allergic sensitization to food proteins and that development of sensitization can be induced by different molecular mechanisms and local immune responses. Our data illustrate the possible contribution of food contaminants in allergic sensitization in humans.
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Ji, Jian; Zhu, Pei; Blaženović, Ivana; Cui, Fangchao; Gholami, Morteza; Sun, Jiadi; Habimana, Jean; Zhang, Yinzhi; Sun, Xiulan
2018-02-28
Urine metabolic profiling of mice was conducted utilizing gas chromatography-mass spectrometry (GC-MS) to investigate the combinatory effect of mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) on the metabolism of the mice. Experiments were conducted by means of five-week-old mice which were individually exposed to 2 mg/kg DON, 20 mg/kg ZEN and the mixture of DON and ZEN (2 mg/kg and 20 mg/kg, respectively). The intragastric administration was applied for three weeks and urine samples were collected for metabolic analysis. Univariate and multivariate analysis were applied to data matrix processing along with respective pathway analysis by MetaMapp and CytoScape. The results showed that the combined DON and ZEN administration resulted in lower significant changes, compared to the individual mycotoxin treated groups verified by heatmap. Metabolic pathways network mapping indicated that the combined mycotoxins treated groups showed a little effect on the metabolites in most pathways, especially in glucose metabolism and its downstream amino acid metabolism. In glucose metabolism, the content of galactose, mannitol, galactonic acid, myo-inositol, tagatose was drastically down-regulated. Furthermore, the organic acids, pyruvate, and amino acids metabolism displayed the same phenomenon. In conclusion, the combined DON/ZEN administration might lead to an "antagonistic effect" in mice metabolism.
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Anorexic action of deoxynivalenol in hypothalamus and intestine.
Tominaga, Misa; Momonaka, Yuka; Yokose, Chihiro; Tadaishi, Miki; Shimizu, Makoto; Yamane, Takumi; Oishi, Yuichi; Kobayashi-Hattori, Kazuo
2016-08-01
Although deoxynivalenol (DON) suppresses food intake and subsequent weight gain, its contribution to anorexia mechanisms has not been fully clarified. Thus, we investigated the anorexic actions of DON in the hypothalamus and intestine, both organs related to appetite. When female B6C3F1 mice were orally exposed to different doses of DON, a drastic anorexic action was observed at a dose of 12.5 mg/kg body weight (bw) from 0 to 3 h after administration. Exposure to DON (12.5 mg/kg bw) for 3 h significantly increased the hypothalamic mRNA levels of anorexic pro-opiomelanocortin (POMC) and its downstream targets, including melanocortin 4 receptor, brain-derived neurotrophic factor, and tyrosine kinase receptor B; at the same time, orexigenic hormones were not affected. In addition, exposure to DON significantly elevated the hypothalamic mRNA levels of proinflammatory cytokines (IL-1β, TNF-α, and IL-6) and activated nuclear factor-kappa B (NF-κB), an upstream factor of POMC. These results suggest that DON-induced proinflammatory cytokines increased the POMC level via NF-κB activation. Moreover, exposure to DON significantly enhanced the gastrointestinal mRNA levels of anorexic cholecystokinin (CCK) and transient receptor potential ankyrin-1 channel (TRPA1), a possible target of DON; these findings suggest that DON induced anorexic action by increasing CCK production via TRPA1. Taken together, these results suggest that DON induces anorexic POMC, perhaps via NF-κB activation, by increasing proinflammatory cytokines in the hypothalamus and brings about CCK production, possibly through increasing intestinal TRPA1 expression, leading to anorexic actions. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Review of mechanisms of deoxynivalenol-induced anorexia: The role of gut microbiota.
Peng, Zhao; Chen, Liangkai; Xiao, Jie; Zhou, Xiaoqi; Nüssler, Andreas K; Liu, Liegang; Liu, Jinping; Yang, Wei
2017-09-01
Deoxynivalenol (DON) is one of the most important mycotoxins in cereal-based foods or other food productions, produced by Fusarium species. Because of the high occurrence of DON in food combined with vast consumption of cereals and grain worldwide, the DON-contaminated food is a very harmful factor for human and animal health. DON has been reported to induce anorexia at lower or chronic doses in animal models. However, further researches for DON-induced anorexia are limited. Previous publications demonstrated a close link between Bacteroidetes and Firmicutes, two kinds of gut microbiota, with weight loss and the effect of low administration of DON on neurotransmitters in the frontal cortex, cerebellum, hypothalamus, hippocampus and pons/medulla. These data are similar to other studies, which show selective 5HT α receptor agonists apparently causing hyperphagia whereas 5HT 1β agonists appear to induce anorexia. Thus, the neurochemical effects of lower DON exposure can be as a result of peripheral toxicological events such as emesis, which overwhelmed its more subtle feed refusal activity. Besides, changes in the microbiota have an impact on stress-related behaviors like anxiety and depression, which can lead to weight loss through decreased feed intake. Gut dysbiosis is also associated with brain dysfunction and with behavioral changes. These conclusions illustrate as well a potential explanation for DON-induced anorexia.In this review, we summarize information about DON-induced anorexia from previous studies and provide our opinion for future investigations that could establish a link between gut microbiota, neurotransmitters, anorexia and weight loss under the DON exposure. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.
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2010-01-01
Background Fusarium head blight is a very important disease of small grain cereals with F. graminearum as one of the most important causal agents. It not only causes reduction in yield and quality but from a human and animal healthcare point of view, it produces mycotoxins such as deoxynivalenol (DON) which can accumulate to toxic levels. Little is known about external triggers influencing DON production. Results In the present work, a combined in vivo/in vitro approach was used to test the effect of sub lethal fungicide treatments on DON production. Using a dilution series of prothioconazole, azoxystrobin and prothioconazole + fluoxastrobin, we demonstrated that sub lethal doses of prothioconazole coincide with an increase in DON production 48 h after fungicide treatment. In an artificial infection trial using wheat plants, the in vitro results of increased DON levels upon sub lethal prothioconazole application were confirmed illustrating the significance of these results from a practical point of view. In addition, further in vitro experiments revealed a timely hyperinduction of H2O2 production as fast as 4 h after amending cultures with prothioconazole. When applying H2O2 directly to germinating conidia, a similar induction of DON-production by F. graminearum was observed. The effect of sub lethal prothioconazole concentrations on DON production completely disappeared when applying catalase together with the fungicide. Conclusions These cumulative results suggest that H2O2 induced by sub lethal doses of the triazole fungicide prothioconazole acts as a trigger of DON biosynthesis. In a broader framework, this work clearly shows that DON production by the plant pathogen F. graminearum is the result of the interaction of fungal genomics and external environmental triggers. PMID:20398299
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Silva, Milena Veronezi; Pante, Giseli Cristina; Romoli, Jéssica Cristina Zoratto; de Souza, Alexandra Perdigão Maia; Rocha, Gustavo Henrique Oliveira da; Ferreira, Flavio Dias; Feijó, Adriane Lettnin Roll; Moscardi, Salesia Maria Prodócimo; de Paula, Karina Ruaro; Bando, Erika; Nerilo, Samuel Botião; Machinski, Miguel
2018-03-01
Deoxynivalenol (DON) is the most important of the trichothecenes in terms of amounts and occurrence in wheat. This compound was shown to be associated with a glomerulonephropathy involving an increase of immunoglobulin A in humans. This study assessed the occurrence of DON in wheat flour and the exposure of Brazilian teenagers, adults and elderly to this mycotoxin due to intake of wheat flour-based products. DON extraction in wheat flour was carried out by solid phase extraction and the quantification was performed by ultra-high proficiency liquid chromatography with diode-array detection. A total of 77.9% of all samples were positive for DON, with concentrations ranging from 73.50 to 2794.63 µg kg -1 . The intake was calculated for the average and 90th percentile of the contamination levels of DON in foods based-wheat for teenagers, adults and elderly in Brazil, and compared with the provisional maximum tolerable daily intakes (PMTDI). Females of all age groups were exposed to DON at higher levels when compared to males in regard of consumption of breads and pastas. Teenagers were the main consumers of foods derived from wheat flour, with maximum probable daily intakes of 1.28 and 1.20 µg kg -1 b.w. day -1 for females and males, respectively. This population is at an increased risk of exposure to DON due to consumption of wheat flour-based foods in Brazil.
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Schwartz-Zimmermann, Heidi E; Hametner, Christian; Nagl, Veronika; Fiby, Iris; Macheiner, Lukas; Winkler, Janine; Dänicke, Sven; Clark, Erica; Pestka, James J; Berthiller, Franz
2017-12-01
The Fusarium mycotoxin deoxynivalenol (DON) is a frequent contaminant of cereal-based food and feed. Mammals metabolize DON by conjugation to glucuronic acid (GlcAc), the extent and regioselectivity of which is species-dependent. So far, only DON-3-glucuronide (DON-3-GlcAc) and DON-15-GlcAc have been unequivocally identified as mammalian DON glucuronides, and DON-7-GlcAc has been proposed as further DON metabolite. In the present work, qualitative HPLC-MS/MS analysis of urine samples of animals treated with DON (rats: 2 mg/kg bw, single bolus, gavage; mice: 1 mg/kg bw, single i.p. injection; pigs: 74 µg/kg bw, single bolus, gavage; cows: 5.2 mg DON/kg dry mass, oral for 13 weeks) revealed additional DON and deepoxy-DON (DOM) glucuronides. To elucidate their structures, DON and DOM were incubated with human (HLM) and rat liver microsomes (RLM). Besides the expected DON/DOM-3- and 15-GlcAc, minor amounts of four DON- and four DOM glucuronides were formed. Isolation and enzymatic hydrolysis of four of these compounds yielded iso-DON and iso-DOM, the identities of which were eventually confirmed by NMR. Incubation of iso-DON and iso-DOM with RLM and HLM yielded two main glucuronides for each parent compound, which were isolated and identified as iso-DON/DOM-3-GlcAc and iso-DON/DOM-8-GlcAc by NMR. Iso-DON-3-GlcAc, most likely misidentified as DON-7-GlcAc in the literature, proved to be a major DON metabolite in rats and a minor metabolite in pigs. In addition, iso-DON-8-GlcAc turned out to be one of the major DON metabolites in mice. DOM-3-GlcAc was the dominant DON metabolite in urine of cows and an important DON metabolite in rat urine. Iso-DOM-3-GlcAc was detected in urine of DON-treated rats and cows. Finally, DON-8,15-hemiketal-8-glucuronide, a previously described by-product of DON-3-GlcAc production by RLM, was identified in urine of DON-exposed mice and rats. The discovery of several novel DON-derived glucuronides in animal urine requires adaptation of the currently used methods for DON-biomarker analysis.
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Fusarium mycotoxin content of UK organic and conventional wheat.
Edwards, S G
2009-04-01
Each year (2001-2005), 300 samples of wheat from fields of known agronomy were analysed for ten trichothecenes by gas chromatography-mass spectrometry (GC/MS) including deoxynivalenol (DON), nivalenol, 3-acetyl-DON, 15-acetyl-DON, fusarenone X, T2 toxin, HT2 toxin, diacetoxyscirpenol, neosolaniol and T-2 triol and zearalenone by high-performance liquid chromatography (HPLC). Of the eleven mycotoxins analysed from 1624 harvest samples of wheat, only eight were detected, and of these only five-deoxynivalenol, 15-acetyl-DON, nivalenol, HT-2 and zearalenone-were detected above 100 microg kg(-1). DON was the most frequently detected Fusarium mycotoxin, present above the limit of quantification (10 microg kg(-1)) in 86% of samples, and was usually present at the highest concentration. The percentage of samples that would have exceeded the recently introduced legal limits varied between 0.4% and 11.3% over the five-year period. There was a good correlation between DON and zearalenone concentrations, although the relative concentration of DON and zearalenone fluctuated between years. Year and region had a significant effect on all mycotoxins analysed. There was no significant difference in the DON concentration of organic and conventional samples. There was also no significant difference in the concentration of zearalenone between organic and conventional samples, however organic samples did have a significantly lower concentration of HT2 and T2. Overall, the risk of UK wheat exceeding the newly introduced legal limits for Fusarium mycotoxins in cereals intended for human consumption is low, but the percentage of samples above these limits will fluctuate between years.
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Antonissen, Gunther; Ducatelle, Richard; Haesebrouck, Freddy; Timbermont, Leen; Verlinden, Marc; Janssens, Geert Paul Jules; Eeckhaut, Venessa; Eeckhout, Mia; De Saeger, Sarah; Hessenberger, Sabine; Martel, An; Croubels, Siska
2014-01-01
Both mycotoxin contamination of feed and Clostridium perfringens-induced necrotic enteritis have an increasing global economic impact on poultry production. Especially the Fusarium mycotoxin deoxynivalenol (DON) is a common feed contaminant. This study aimed at examining the predisposing effect of DON on the development of necrotic enteritis in broiler chickens. An experimental Clostridium perfringens infection study revealed that DON, at a contamination level of 3,000 to 4,000 µg/kg feed, increased the percentage of birds with subclinical necrotic enteritis from 20±2.6% to 47±3.0% (P<0.001). DON significantly reduced the transepithelial electrical resistance in duodenal segments (P<0.001) and decreased duodenal villus height (P = 0.014) indicating intestinal barrier disruption and intestinal epithelial damage, respectively. This may lead to an increased permeability of the intestinal epithelium and decreased absorption of dietary proteins. Protein analysis of duodenal content indeed showed that DON contamination resulted in a significant increase in total protein concentration (P = 0.023). Furthermore, DON had no effect on in vitro growth, alpha toxin production and netB toxin transcription of Clostridium perfringens. In conclusion, feed contamination with DON at concentrations below the European maximum guidance level of 5,000 µg/kg feed, is a predisposing factor for the development of necrotic enteritis in broilers. These results are associated with a negative effect of DON on the intestinal barrier function and increased intestinal protein availability, which may stimulate growth and toxin production of Clostridium perfringens. PMID:25268498
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Hewitt, M A; Girgis, G N; Brash, M; Smith, T K
2012-12-01
The objective of the present study was to determine the effects of feeding diets containing grains naturally contaminated with Fusarium mycotoxins to fryer rabbits. The efficacy of a glucomannan mycotoxin adsorbent (GMA) was also examined. Thirty 5-wk-old male New Zealand White rabbits were fed a control diet, a contaminated diet, or a contaminated diet + 0.2% GMA for 21 d. Experimental diets contained deoxynivalenol (DON; vomitoxin) at a concentration of 0.25, 4.3, and 4.9 μg/g, respectively. Feed intake was measured daily and water intake was measured every 3 d. At the end of the feeding period, blood was collected for evaluation of serum chemistry and hematology. Visceral organs were excised, weighed, and processed for histopathological examination. Body weight gain and water intake were greater in rabbits fed the contaminated diet (P = 0.075 and 0.020, respectively) and those fed the contaminated + GMA diet (P = 0.026 and 0.002, respectively) compared with controls. Rabbits fed the contaminated + GMA diet had significantly increased serum urea concentrations (P = 0.023) and decreased serum alkaline phosphatase activity (P = 0.020) compared with controls. Increase in BW gain of rabbits fed the contaminated diets was caused by increased water consumption. There was no effect (P > 0.05) of diet on relative organ weights, but decreased infiltrations with eosinophilic granulocytes were observed in different regions of the intestine in rabbits fed the contaminated or the contaminated + GMA diet. It was concluded that rabbits could be adversely affected by feed-borne Fusarium mycotoxins but appear to be less sensitive than other mammalian species. Supplementation with GMA did not reduce many of the effects of feed-borne mycotoxins.
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Deoxynivalenol, zearalenone and T-2 in grain based swine feed in Hungary.
Tima, Helga; Rácz, Anita; Guld, Zsuzsanna; Mohácsi-Farkas, Csilla; Kiskó, Gabriella
2016-12-01
Fusarium genera can produce trichothecenes like deoxynivalenol (DON), zearalenone (ZEN) and T-2 toxin, which can occur in feed cereal grains. Enzyme-linked immunosorbent assays (ELISA) tests of different Hungarian swine feedstuff proved that these mycotoxins were present. In this survey, 45 feed samples from 3 significant Hungarian swine feedstuff manufacturers were tested. ELISA methodology validation showed mean recovery rates in ranges from 85.3% to 98.1%, with intermediate precision of 86.9-96.9% and variation coefficients of 3.4-5.7% and 5.9-7.1%, respectively. The results showed that among Fusarium toxins, generally DON was present in the highest concentration, followed by T-2 and finally ZEN in all tested swine feeds. Each of the mycotoxins was found above the limit of detection in all swine feedstuffs. Boars feed's DON (average ± standard deviation was 872 ± 139 µg kg -1 ) and ZEN (172 ± 18 µg kg -1 ) results of one of the manufacturers were above the guidance values. It indicates the necessity for efficient monitoring of DON, ZEN and T-2 mycotoxins in swine feeds.
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Pasternak, J Alex; Aiyer, Vaishnavi Iyer Aka; Hamonic, Glenn; Beaulieu, A Denise; Columbus, Daniel A; Wilson, Heather L
2018-01-12
We intended to assess how exposure of piglets to deoxynivalenol (DON)-contaminated feed impacted their growth, immune response and gut development. Piglets were fed traditional Phase I, Phase II and Phase III diets with the control group receiving 0.20-0.40 ppm DON (referred to as the Control group) and treatment group receiving much higher level of DON-contaminated wheat (3.30-3.80 ppm; referred to as DON-contaminated group). Feeding a DON-contaminated diet had no impact on average daily feed intake (ADFI) ( p < 0.08) or average daily gain (ADG) ( p > 0.10) but it did significantly reduce body weight over time relative to the control piglets ( p < 0.05). Cytokine analysis after initial exposure to the DON-contaminated feed did not result in significant differences in serum interleukin (IL) IL1β, IL-8, IL-13, tumor necrosis factor (TNF)-α or interferon (IFN)-γ. After day 24, no obvious changes in jejunum or ileum gut morphology, histology or changes in gene expression for IL-1β, IL-6, IL-10, TNFα, or Toll-like receptor (TLR)-4 genes. IL-8 showed a trend towards increased expression in the ileum in DON-fed piglets. A significant increase in gene expression for claudin (CLDN) 7 gene expression and a trend towards increased CLDN 2-expression was observed in the ileum in piglets fed the highly DON-contaminated wheat. Because CLDN localization was not negatively affected, we believe that it is unlikely that gut permeability was affected. Exposure to DON-contaminated feed did not significantly impact weaner piglet performance or gut physiology.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Islam, Zahidul; Center for Integrative Toxicology, Michigan State University, 234 G.M. Trout Building, Michigan State University, East Lansing, MI 48824-1224; Gray, Jennifer S.
2006-06-15
The effects of the ribotoxic trichothecene deoxynivalenol (DON) on mitogen-activated protein kinase (MAPK)-mediated IL-8 expression were investigated in cloned human monocytes and peripheral blood mononuclear cells (PBMC). DON (250 to 1000 ng/ml) induced both IL-8 mRNA and IL-8 heteronuclear RNA (hnRNA), an indicator of IL-8 transcription, in the human U937 monocytic cell line in a concentration-dependent manner. Expression of IL-8 hnRNA, mRNA and protein correlated with p38 phosphorylation and was completely abrogated by the p38 MAPK inhibitor SB203580. DON at 500 ng/ml similarly induced p38-dependent IL-8 protein and mRNA expression in PBMC cultures from healthy volunteers. Significantly increased IL-6 andmore » IL-1{beta} intracellular protein and mRNA expression was also observed in PBMC treated with DON (500 ng/ml) which were also partially p38-dependent. Flow cytometry of PBMC revealed that DON-induced p38 phosphorylation varied among individuals relative to both threshold toxin concentrations (25-100 ng/ml) and relative increases in percentages of phospho-p38{sup +} cells. DON-induced p38 activation occurred exclusively in the CD14{sup +} monocyte population. DON was devoid of agonist activity for human Toll-like receptors 2, 3, 4, 5, 7, 8 and 9. However, two other ribotoxins, emetine and anisomycin, induced p38 phosphorylation in PBMC similarly to DON. Taken together, these data suggest that (1) p38 activation was required for induction of IL-8 and proinflammatory gene expression in the monocyte and (2) DON induced p38 activation in human monocytes via the ribotoxic stress response.« less
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The effects of deoxynivalenol on gene expression in the murine thymus
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kol, Sandra W.M. van; Department of Health Risk Analysis and Toxicology, Maastricht University; Netherlands Toxicogenomics Centre
Deoxynivalenol (DON) is a mycotoxin produced by several Fusarium species and is often detected in grains. Because of its high abundance, there has been a large interest in the effects of DON in animals and humans. DON is known to be immunosuppressive at high concentrations and immunostimulatory at low concentrations. The present study aimed to acquire insight into the modes of action of DON. For this, C57Bl6 mice were orally exposed to 5, 10, or 25 mg/kg bw DON for 3, 6, or 24 h and thymuses were subjected to genome-wide expression microarray analysis. Gene set enrichment analysis (GSEA) demonstratedmore » that DON downregulated genes involved in proliferation, mitochondria, protein synthesis, and ribosomal proteins. Furthermore, GSEA showed a selective downregulation of genes highly expressed at the early precursor thymocytes stage. This indicates that early precursor thymocytes, particularly at the double-positive CD4+CD8+ stage, are more vulnerable to DON than very early or late precursor thymocytes. There was a large overlap of genes upregulated by DON with genes previously reported to be either upregulated during T cell activation or upregulated during negative selection of thymocytes that recognize 'self-antigens'. This indicates that DON induces cellular events that also occur after activation of the T cell receptor, for example, release of calcium from the endoplasmatic reticulum. This T cell activation in the thymus then evokes negative selection and depletion of thymocytes, which provides a plausible explanation for the high sensitivity of the thymus for DON exposure. The expression patterns of four genes indicative for some of the processes that were affected after DON treatment were confirmed using real-time PCR. Immunocytological experiments with primary mouse thymocytes demonstrated the translocation of NFAT from the cytoplasm into the nucleus upon exposure top DON, thus providing further evidence for the involvement of T cell activation.« less
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2011-01-01
Background The trichothecene mycotoxin deoxynivalenol (DON) may be concentrated in distillers dried grains with solubles (DDGS; a co-product of fuel ethanol fermentation) when grain containing DON is used to produce fuel ethanol. Even low levels of DON (≤ 5 ppm) in DDGS sold as feed pose a significant threat to the health of monogastric animals. New and improved strategies to reduce DON in DDGS need to be developed and implemented to address this problem. Enzymes known as trichothecene 3-O-acetyltransferases convert DON to 3-acetyldeoxynivalenol (3ADON), and may reduce its toxicity in plants and animals. Results Two Fusarium trichothecene 3-O-acetyltransferases (FgTRI101 and FfTRI201) were cloned and expressed in yeast (Saccharomyces cerevisiae) during a series of small-scale ethanol fermentations using barley (Hordeum vulgare). DON was concentrated 1.6 to 8.2 times in DDGS compared with the starting ground grain. During the fermentation process, FgTRI101 converted 9.2% to 55.3% of the DON to 3ADON, resulting in DDGS with reductions in DON and increases in 3ADON in the Virginia winter barley cultivars Eve, Thoroughbred and Price, and the experimental line VA06H-25. Analysis of barley mashes prepared from the barley line VA04B-125 showed that yeast expressing FfTRI201 were more effective at acetylating DON than those expressing FgTRI101; DON conversion for FfTRI201 ranged from 26.1% to 28.3%, whereas DON conversion for FgTRI101 ranged from 18.3% to 21.8% in VA04B-125 mashes. Ethanol yields were highest with the industrial yeast strain Ethanol Red®, which also consumed galactose when present in the mash. Conclusions This study demonstrates the potential of using yeast expressing a trichothecene 3-O-acetyltransferase to modify DON during commercial fuel ethanol fermentation. PMID:21888629
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Ghareeb, Khaled; Awad, Wageha A.; Sid-Ahmed, Omer E.; Böhm, Josef
2014-01-01
Mycotoxins pose an important danger to human and animal health. Poultry feeds are frequently contaminated with deoxynivalenol (DON) mycotoxin. It is thus of great importance to evaluate the effects of DON on the welfare related parameters in poultry industry. In the present study, the effects of contamination of broiler diet with 10 mg DON/kg feed on plasma corticosterone and heterophil to lymphocyte (H/L) ratio as indicators of stress, tonic immobility duration as an index for fear response and growth performance of broiler chickens were studied. In addition, the effect of a microbial feed additive either alone or in combination with DON contamination on these different aspects was also evaluated. The results showed that DON feeding significantly affected the welfare related parameters of broiler chickens. The feeding of DON contaminated diet resulted in an elevation of plasma corticosterone, higher H/L ratio and increased the fear levels as indicated by longer duration of tonic immobility reaction. Furthermore, DON reduced the body weight and body weight gain during the starter phase definitely at the second and third week. However, during grower phase, feeding of DON decreased the body weight at the fourth week and reduced the body gain at the fifth week. Addition of the microbial feed additive, a commercial antidote for DON mycotoxin, was able to overcome DON effects on stress index (H/L ratio), fearfulness and growth parameters of broilers. In conclusion, we showed for the first time that the DON feeding increased the underlying fearfulness and physiological stress responses of broilers and resulted in a reduction in the welfare status as indicated by higher plasma corticosterone, higher H/L ratio and higher fearfulness. Additionally, feeding the microbial feed additive was effective in reducing the adverse effects of DON on the bird's welfare and can improve the performance of broiler chickens. PMID:24498179
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Choi, Byung-Kook; Jeong, Sang-Hee; Cho, Joon-Hyung; Shin, Hyo-Sook; Son, Seong-Wan; Yeo, Young-Keun; Kang, Hwan-Goo
2013-08-01
Mice were exposed to deoxynivalenol (DON) via drinking water at a concentration of 2 mg/L for 36 days. On day 8 of treatment, inactivated porcine parvovirus vaccine (PPV) was injected intraperitoneally. The relative and absolute weight of the spleen was significantly decreased in the DON-treated group (DON). Antibody titers to parvovirus in serum were 47.9 ± 2.4 in the vaccination group (Vac), but 15.2 ± 6.5 in the group treated with DON and vaccine (DON + Vac). The IgA and IgG was not different in the DON, Vac an,d DON + Vac groups. IgM was significantly lower only in the DON + Vac group. However IgE was significantly increased in the Vac and DON + Vac group, but no change was observed between the Vac and DON + Vac groups. The concentrations of IL-2, IL-4, GM-CSF, MCP-1 and Rantes in serum, and IL-1α in mesenteric lymph node and MIP-1β in spleen were significantly increased by DON treatment compared to control. The concentrations of IL-2, IL-5, IL-6, IL-9, IL-12, IL-13 and Rantes in thymus, of IL-2 in spleen, and of IL-1α, IL-1β, IL-3, IL-5, IL-10, IL-17, G-CSF, GM-CSF and MCP-1 in mesenteric lymph nodes were significantly decreased in mice compared to those in the Vac group, while concentrations of IL-1α, IL-2, IL-9, IL-13,G-CSF, GM-CSF, IFN-γ, MCP-1, MIP-1α and TNF-α were significantly increased in serum compared to the Vac group. In conclusion, the results presented here indicate that exposure to DON at 2.0 mg/L via drinking water can disrupt the immune response in vaccinated mice by modulating cytokines and chemokines involved in their immune response to infectious disease.
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Ghareeb, Khaled; Awad, Wageha A; Sid-Ahmed, Omer E; Böhm, Josef
2014-01-01
Mycotoxins pose an important danger to human and animal health. Poultry feeds are frequently contaminated with deoxynivalenol (DON) mycotoxin. It is thus of great importance to evaluate the effects of DON on the welfare related parameters in poultry industry. In the present study, the effects of contamination of broiler diet with 10 mg DON/kg feed on plasma corticosterone and heterophil to lymphocyte (H/L) ratio as indicators of stress, tonic immobility duration as an index for fear response and growth performance of broiler chickens were studied. In addition, the effect of a microbial feed additive either alone or in combination with DON contamination on these different aspects was also evaluated. The results showed that DON feeding significantly affected the welfare related parameters of broiler chickens. The feeding of DON contaminated diet resulted in an elevation of plasma corticosterone, higher H/L ratio and increased the fear levels as indicated by longer duration of tonic immobility reaction. Furthermore, DON reduced the body weight and body weight gain during the starter phase definitely at the second and third week. However, during grower phase, feeding of DON decreased the body weight at the fourth week and reduced the body gain at the fifth week. Addition of the microbial feed additive, a commercial antidote for DON mycotoxin, was able to overcome DON effects on stress index (H/L ratio), fearfulness and growth parameters of broilers. In conclusion, we showed for the first time that the DON feeding increased the underlying fearfulness and physiological stress responses of broilers and resulted in a reduction in the welfare status as indicated by higher plasma corticosterone, higher H/L ratio and higher fearfulness. Additionally, feeding the microbial feed additive was effective in reducing the adverse effects of DON on the bird's welfare and can improve the performance of broiler chickens.
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In vivo toxicity assessment of deoxynivalenol-contaminated wheat after ozone degradation.
Wang, Li; Wang, Ying; Shao, Huili; Luo, Xiaohu; Wang, Ren; Li, Yongfu; Li, Yanan; Luo, Yingpeng; Zhang, Dongjie; Chen, Zhengxing
2017-01-01
The effect of ozone on deoxynivalenol (DON) detoxification was investigated. Ozone treatment could significantly reduce the levels of DON in wheat; 53% of DON in wheat was decomposed with 90 mg l -1 of ozone at a flow rate of 15 l min -1 for 4 h. The safety of DON-contaminated wheats (DCWs) untreated/treated by ozone was also evaluated. Institute of Cancer Research (ICR) mice were divided into a standard diet group and five experimental diet groups for a 51-day orally administration experiment. In the experiment, no remarkable changes in the general appearance of the mice were observed, and all the mice survived until the scheduled necropsy. The results of sub-chronic toxicity indicated that mice fed on DCWs alone had significantly decreased in body weight gain, thymus and spleen weights, ratios of liver, thymus and spleen to body weight, blood indices (red blood cell, haemoglobin, white blood cell), and pro-inflammatory cytokines (interleukin-6 (IL-6), tumour necrosis factor alpha (TNF-α)), while showing a significant increase in alanine aminotransferase, aspartate aminotransferase, blood creatinine and blood urea nitrogen levels. Histopathological examination indicate that DON elicited some degree of toxicity on the liver, kidney and thymus tissue. Mice fed on DCWs treated by ozone mitigated the adverse effects compared with mice fed on DCWs. All the results suggested that the deleterious effects of DON could be highly reduced by ozone, and ozone itself shows minor toxic effects on animals in this process.
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Szabó-Fodor, Judit; Fébel, Hedvig; Mézes, Miklós; Balogh, Krisztián; Bázár, György; Kocsó, Dániel; Kovács, Melinda
2017-01-01
(1) Background and (2) Methods: A 14-day in vivo, multitoxic (pure mycotoxins) rat experiment was conducted with zearalenone (ZEA; 15 μg/animal/day), deoxynivalenol (DON; 30 μg/animal/day) and fumonisin B1 (FB1; 150 μg/animal/day), as individual mycotoxins, binary (FD, FZ and DZ) and ternary combinations (FDZ), via gavage in 1 mL water boluses. (3) Results: Body weight was unaffected, while liver (ZEA↑ vs. DON) and kidney weight (ZEA↑ vs. FDZ) increased. Hepatocellular membrane lipid fatty acids (FAs) referred to ceramide synthesis disturbance (C20:0, C22:0), and decreased unsaturation (C22:5 n3 and unsat. index), mainly induced by DON and to a lesser extent by ZEA. The DON-FB1 interaction was additive on C20:0 in liver lipids. In renal phospholipids, ZEA had the strongest effect on the FA profile, affecting the saturated (C18:0) and many n6 FAs; ZEA was in an antagonistic relationship with FB1 (C18:0) or DON (C18:2 n6, C20:1 n9). Hepatic oxidative stress was the most expressed in FD (reduced glutathione and glutathione peroxidase), while the nephrotoxic effect was further supported by lipid peroxidation (malondialdehyde) in the DON treatment. (4) Conclusions: In vivo study results refer to multiple mycotoxin interactions on membrane FAs, antioxidants and lipid peroxidation compounds, needing further testing. PMID:29271890
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Ren, Zhihua; Wang, Yachao; Deng, Huidan; Deng, Youtian; Deng, Junliang; Zuo, Zhicai; Wang, Ya; Peng, Xi; Cui, Hengmin; Shen, Liuhong; Ma, Xiaoping; Fang, Jing
2015-04-01
Deoxynivalenol (DON) immunotoxicity and its induction of cytokines and related genes in the splenic lymphocytes of chickens have not been completely elucidated. In the present study, we aimed to evaluate the effects of 48 h of different DON treatments (0 μg/mL, 0.2 μg/mL, 0.8 μg/mL, 3.2 μg/mL, 12.5 μg/mL, and 50 μg/mL) on the secretion and the mRNA expressions of some cytokine genes, such as interleukin-1beta (IL-1β), IL-1RI, IL-2, IL-4, IL-6, IL-10, IL-12β, and IFN-γ in chicken splenic lymphocytes. The concentrations of IL-1RI, IL-6, IL-10, and IFN-γ were increased with the DON concentrations increasing (P<0.05 or P<0.01). However, the concentrations of IL-1β, IL-2, IL-4, and IL-12β were decreased with the DON concentrations increasing (P<0.05 or P<0.01). Except IL-1β, the mRNA expressions of the other cytokines were up-regulated by DON. The highest mRNA expressions values of IL-1RI, IL-4, IL-10, IL-12β, and IFN-γ were at 50 μg/mL DON treatment groups (P<0.05 or P<0.01), while the highest mRNA expressions values of IL-2 and IL-6 were at 12.5 μg/mL DON treatment groups (P<0.05 or P<0.01). Our data revealed that the potent effects of DON in affecting the secretion and the mRNA expression of the related cytokines in chicken splenic lymphocytes in vitro. Copyright © 2014 Elsevier Ltd. All rights reserved.
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Alassane-Kpembi, Imourana; Pinton, Philippe; Hupé, Jean-François; Neves, Manon; Lippi, Yannick; Combes, Sylvie; Castex, Mathieu; Oswald, Isabelle P
2018-05-15
Type B trichothecene mycotoxin deoxynivalenol (DON) is one of the most frequently occurring food contaminants. By inducing trans-activation of a number of pro-inflammatory cytokines and increasing the stability of their mRNA, trichothecene can impair intestinal health. Several yeast products, especially Saccharomyces cerevisiae , have the potential for improving the enteric health of piglets, but little is known about the mechanisms by which the administration of yeast counteracts the DON-induced intestinal alterations. Using a pig jejunum explant model, a whole-transcriptome analysis was performed to decipher the early response of the small intestine to the deleterious effects of DON after administration of S. cerevisiae boulardii strain CNCM I-1079. Compared to the control condition, no differentially expressed gene (DE) was observed after treatment by yeast only. By contrast, 3619 probes-corresponding to 2771 genes-were differentially expressed following exposure to DON, and 32 signaling pathways were identified from the IPA software functional analysis of the set of DE genes. When the intestinal explants were treated with S. cerevisiae boulardii prior to DON exposure, the number of DE genes decreased by half (1718 probes corresponding to 1384 genes). Prototypical inflammation signaling pathways triggered by DON, including NF-κB and p38 MAPK, were reversed, although the yeast demonstrated limited efficacy toward some other pathways. S. cerevisiae boulardii also restored the lipid metabolism signaling pathway, and reversed the down-regulation of the antioxidant action of vitamin C signaling pathway. The latter effect could reduce the burden of DON-induced oxidative stress. Altogether, the results show that S. cerevisiae boulardii reduces the DON-induced alteration of intestinal transcriptome, and point to new mechanisms for the healing of tissue injury by yeast.
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Alassane-Kpembi, Imourana; Hupé, Jean-François; Neves, Manon; Lippi, Yannick; Combes, Sylvie; Castex, Mathieu
2018-01-01
Type B trichothecene mycotoxin deoxynivalenol (DON) is one of the most frequently occurring food contaminants. By inducing trans-activation of a number of pro-inflammatory cytokines and increasing the stability of their mRNA, trichothecene can impair intestinal health. Several yeast products, especially Saccharomyces cerevisiae, have the potential for improving the enteric health of piglets, but little is known about the mechanisms by which the administration of yeast counteracts the DON-induced intestinal alterations. Using a pig jejunum explant model, a whole-transcriptome analysis was performed to decipher the early response of the small intestine to the deleterious effects of DON after administration of S. cerevisiae boulardii strain CNCM I-1079. Compared to the control condition, no differentially expressed gene (DE) was observed after treatment by yeast only. By contrast, 3619 probes—corresponding to 2771 genes—were differentially expressed following exposure to DON, and 32 signaling pathways were identified from the IPA software functional analysis of the set of DE genes. When the intestinal explants were treated with S. cerevisiae boulardii prior to DON exposure, the number of DE genes decreased by half (1718 probes corresponding to 1384 genes). Prototypical inflammation signaling pathways triggered by DON, including NF-κB and p38 MAPK, were reversed, although the yeast demonstrated limited efficacy toward some other pathways. S. cerevisiae boulardii also restored the lipid metabolism signaling pathway, and reversed the down-regulation of the antioxidant action of vitamin C signaling pathway. The latter effect could reduce the burden of DON-induced oxidative stress. Altogether, the results show that S. cerevisiae boulardii reduces the DON-induced alteration of intestinal transcriptome, and point to new mechanisms for the healing of tissue injury by yeast. PMID:29762474
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Uhlig, Silvio; Stanic, Ana; Hofgaard, Ingerd S.; Kluger, Bernhard; Schuhmacher, Rainer; Miles, Christopher O.
2016-01-01
A glutathione (GSH) adduct of the mycotoxin 4-deoxynivalenol (DON), together with a range of related conjugates, has recently been tentatively identified by LC-MS of DON-treated wheat spikelets. In this study, we prepared samples of DON conjugated at the 10- and 13-positions with GSH, Cys, CysGly, γ-GluCys and N-acetylcysteine (NAC). The mixtures of conjugates were used as standards for LC-HRMS analysis of one of the DON-treated wheat spikelet samples, as well as 19 Norwegian grain samples of spring wheat and 16 grain samples of oats that were naturally-contaminated with DON at concentrations higher than 1 mg/kg. The artificially-contaminated wheat spikelets contained conjugates of GSH, CysGly and Cys coupled at the olefinic 10-position of DON, whereas the naturally-contaminated harvest-ripe grain samples contained GSH, CysGly, Cys, and NAC coupled mainly at the 13-position on the epoxy group. The identities of the conjugates were confirmed by LC-HRMS comparison with authentic standards, oxidation to the sulfoxides with hydrogen peroxide, and examination of product-ion spectra from LC-HRMS/MS analysis. No γ-GluCys adducts of DON were detected in any of the samples. The presence of 15-O-acetyl-DON was demonstrated for the first time in Norwegian grain. The results indicate that a small but significant proportion of DON is metabolized via the GSH-conjugation pathway in plants. To our knowledge, this is the first report of in vivo conjugation of trichothecenes via their epoxy group, which has generally been viewed as unreactive. Because conjugation at the 13-position of DON and other trichothecenes has been shown to be irreversible, this type of conjugate may prove useful as a biomarker of exposure to DON and other 12,13-epoxytrichothecenes. PMID:27845722
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Ji, G E; Park, S Y; Wong, S S; Pestka, J J
1998-02-06
Characterization of how vomitoxin (VT) and other trichothecenes affect macrophage regulatory and effector function may contribute to improved understanding of mechanisms by which these mycotoxins impact the immune system. The RAW 264.7 murine cell line was used as a macrophage model to assess effects of the VT on proliferation and the production of nitric oxide (NO), hydrogen peroxide (H2O2) and cytokines. Using the MTT cleavage assay, VT at concentrations of 50 ng/ml or higher was found to significantly decrease proliferation and viability of RAW 264.7 cells without stimulation or with stimulation by lipopolysaccharide (LPS) or interferon (IFN)-gamma. In the absence of an activation agent, VT (25-250 ng/ml) had negligible effects on the production of NO, H2O2, and cytokines. Upon activation with LPS at concentrations of 10 to 100 ng/ml, VT at 25-100 ng/ml markedly enhanced production of H2O2 but was inhibitory at 250 ng/ml. VT enhancement of H2O2 production was observed as early as 12 h after LPS stimulation. When IFN-gamma was used as the stimulant, VT (25-250 ng/ml) delayed peak H2O2 production. VT (25-250 ng/ml) also markedly decreased NO production in cells activated with LPS or IFN-gamma. Interestingly, VT superinduced TNF-alpha and IL-6 production in LPS-stimulated cells and also elevated TNF-alpha in IFN-gamma stimulated cells. These results suggest that VT can selectively and concurrently upregulate or downregulate critical functions associated with activated macrophages.
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Deoxynivalenol induced mouse skin cell proliferation and inflammation via MAPK pathway
DOE Office of Scientific and Technical Information (OSTI.GOV)
Mishra, Sakshi; Department of Biochemistry, Banaras Hindu University; Tripathi, Anurag
Several toxicological manifestations of deoxynivalenol (DON), a mycotoxin, are well documented; however, dermal toxicity is not yet explored. The effect of topical application of DON to mice was studied using markers of skin proliferation, inflammation and tumor promotion. Single topical application of DON (84–672 nmol/mouse) significantly enhanced dermal hyperplasia and skin edema. DON (336 and 672 nmol) caused significant enhancement in [{sup 3}H]-thymidine uptake in DNA along with increased myeloperoxidase and ornithine decarboxylase activities, suggesting tissue inflammation and cell proliferation. Furthermore, DON (168 nmol) caused enhanced expression of RAS, and phosphorylation of PI3K/Akt, ERK, JNK and p38 MAPKs. DON exposuremore » also showed activation of transcription factors, c-fos, c-jun and NF-κB along with phosphorylation of IkBα. Enhanced phosphorylation of NF-κB by DON caused over expression of target proteins, COX-2, cyclin D1 and iNOS in skin. Though a single topical application of DMBA followed by twice weekly application of DON (84 and 168 nmol) showed no tumorigenesis after 24 weeks, however, histopathological studies suggested hyperplasia of the epidermis and hypertrophy of hair follicles. Interestingly, intestine was also found to be affected as enlarged Peyer's patches were observed, suggesting inflammatory effects which were supported by elevation of inflammatory cytokines after 24 weeks of topical application of DON. These results suggest that DON induced cell proliferation in mouse skin is through the activation of MAPK signaling pathway involving transcription factors NFκB and AP-1, further leading to transcriptional activation of downstream target proteins c-fos, c-jun, cyclin D1, iNOS and COX-2 which might be responsible for its inflammatory potential. - Highlights: • Topical application of DON enhanced epidermal inflammation and cell proliferation. • DON follows PI3K/Akt/MAPK signaling cascade, with activation of AP-1 and NF-κB. • DON caused over expression of target proteins, COX-2, cyclin D1 and iNOS in skin. • No tumor promotion was observed up to 24 weeks of topical application of DON. • Enhanced Peyer's patches and inflammatory cytokines suggested inflammation in skin.« less
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Awad, Wageha A.; Ghareeb, Khaled; Dadak, Agnes; Hess, Michael; Böhm, Josef
2014-01-01
The immune and intestinal epithelial cells are particularly sensitive to the toxic effects of deoxynivalenol (DON). The aim of this experiment was to study the effects of DON and/or a microbial feed additive on the DNA damage of blood lymphocytes and on the level of thiobarbituric acid reactive substance (TBARS) as an indicator of lipid peroxidation and oxidative stress in broilers. A total of forty 1-d-old broiler chicks were randomly assigned to 1 of 4 dietary treatments (10 birds per group) for 5 wk. The dietary treatments were 1) basal diet; 2) basal diet contaminated with 10 mg DON/kg feed; 3) basal diet contaminated with 10 mg DON/kg feed and supplemented with 2.5 kg/ton of feed of Mycofix Select; 4) basal diet supplemented with Mycofix Select (2.5 kg/ton of feed). At the end of the feeding trial, blood were collected for measuring the level of lymphocyte DNA damage of blood and the TBARS level was measured in plasma, heart, kidney, duodenum and jejunum. The dietary exposure of DON caused a significant increase (P = 0.001) of DNA damage in blood lymphocytes (31.99±0.89%) as indicated in the tail of comet assay. Interestingly addition of Mycofix Select to DON contaminated diet decreased (P = 0.001) the DNA damage (19.82±1.75%) induced by DON. In order to clarify the involvement of lipid peroxidation in the DNA damage of DON, TBARS levels was measured. A significant increase (P = 0.001) in the level of TBARS (23±2 nmol/mg) was observed in the jejunal tissue suggesting that the lipid peroxidation might be involved in the DNA damage. The results indicate that DON is cytotoxic and genotoxic to the chicken intestinal and immune cells and the feed additive have potential ability to prevent DNA damage induced by DON. PMID:24498242
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Stereoselective Luche reduction of deoxynivalenol and three of its acetylated derivatives at C8.
Fruhmann, Philipp; Hametner, Christian; Mikula, Hannes; Adam, Gerhard; Krska, Rudolf; Fröhlich, Johannes
2014-01-10
The trichothecene mycotoxin deoxynivalenol (DON) is a well known and common contaminant in food and feed. Acetylated derivatives and other biosynthetic precursors can occur together with the main toxin. A key biosynthetic step towards DON involves an oxidation of the 8-OH group of 7,8-dihydroxycalonectrin. Since analytical standards for the intermediates are not available and these intermediates are therefore rarely studied, we aimed for a synthetic method to invert this reaction, making a series of calonectrin-derived precursors accessible. We did this by developing an efficient protocol for stereoselective Luche reduction at C8. This method was used to access 3,7,8,15-tetrahydroxyscirpene, 3-deacetyl-7,8-dihydroxycalonectrin, 15-deacetyl-7,8-dihydroxycalonectrin and 7,8-dihydroxycalonectrin, which were characterized using several NMR techniques. Beside the development of a method which could basically be used for all type B trichothecenes, we opened a synthetic route towards different acetylated calonectrins.
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Flannery, Brenna M.; Amuzie, Chidozie J.; Pestka, James J.
2013-01-01
Consumption of the trichothecene deoxynivalenol (DON) suppresses growth in experimental animals - an adverse effect that was used to establish the tolerable daily intake for this toxin. DON ingestion has been recently found to suppress plasma insulin-like growth factor acid-labile subunit (IGFALS), a protein essential for growth. Studies were conducted to explore the feasibility of using plasma IGFALS as a biomarker of effect for DON. In the first study, weanling mice were fed 0, 1, 2.5, 5 and 10 ppm DON and weight and plasma IGFALS determined at intervals over 9 wk. Reduced body weight gains were detectable beginning at wk 5 in the 10 ppm dose and wk 7 at the 5 ppm dose. Plasma IGFALS was significantly depressed at wk 5 in the 5 and 10 ppm groups at wk 9 in the 10 ppm group. Depressed IGFALS significantly correlated with reduced body weight at wk 5 and 9. Benchmark dose modeling revealed the BMDL and BMD for plasma IGFALS reduction were 1.1 and 3.0 ppm DON and for weight reduction were 2.1 and 4.5 ppm DON. In the second study, it was demonstrated that mice fed 15 ppm DON diet had significantly less plasma IGFALS than mice fed identical amounts of control diet. Thus DON’s influence on IGFALS likely reflects the combined effects of reduced food intake as well as its physiological action involving suppressors of cytokine signaling. Taken together, these findings suggest that plasma IGFALS might be a useful biomarker for DON’s adverse effects on growth. PMID:23298694
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De Girolamo, A; Lippolis, V; Nordkvist, E; Visconti, A
2009-06-01
Fourier transform near-infrared spectroscopy (FT-NIR) was used for rapid and non-invasive analysis of deoxynivalenol (DON) in durum and common wheat. The relevance of using ground wheat samples with a homogeneous particle size distribution to minimize measurement variations and avoid DON segregation among particles of different sizes was established. Calibration models for durum wheat, common wheat and durum + common wheat samples, with particle size <500 microm, were obtained by using partial least squares (PLS) regression with an external validation technique. Values of root mean square error of prediction (RMSEP, 306-379 microg kg(-1)) were comparable and not too far from values of root mean square error of cross-validation (RMSECV, 470-555 microg kg(-1)). Coefficients of determination (r(2)) indicated an "approximate to good" level of prediction of the DON content by FT-NIR spectroscopy in the PLS calibration models (r(2) = 0.71-0.83), and a "good" discrimination between low and high DON contents in the PLS validation models (r(2) = 0.58-0.63). A "limited to good" practical utility of the models was ascertained by range error ratio (RER) values higher than 6. A qualitative model, based on 197 calibration samples, was developed to discriminate between blank and naturally contaminated wheat samples by setting a cut-off at 300 microg kg(-1) DON to separate the two classes. The model correctly classified 69% of the 65 validation samples with most misclassified samples (16 of 20) showing DON contamination levels quite close to the cut-off level. These findings suggest that FT-NIR analysis is suitable for the determination of DON in unprocessed wheat at levels far below the maximum permitted limits set by the European Commission.
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Pascale, Michelangelo; De Girolamo, Annalisa; Visconti, Angelo; Magan, Naresh; Chianella, Iva; Piletska, Elena V; Piletsky, Sergey A
2008-02-25
Molecular modelling and computational design were used to identify itaconic acid (IA) as a functional monomer with high affinity towards deoxynivalenol (DON), a Fusarium-toxin frequently occurring in cereals. IA-based polymers were photochemically synthesised in dimethyl formamide (porogen) using ethylenglycol dimethacrylate as cross-linker and 1,1'-azo-bis(cyclohexane carbonitrile) as initiator, and the relevant binding interactions with DON in solvents with different polarity were investigated. The performances of the non-imprinted IA-based polymer (blank polymer, BP) and the corresponding molecularly imprinted polymer (MIP) were compared using DON as a template. Both BP and MIP were able to bind about 90% DON either in toluene, water or water containing 5% polyethylene glycol. Non-imprinted polymers with different molar ratios of IA to cross-linker were evaluated as adsorbents for solid-phase extraction (SPE) clean-up and pre-concentration of DON from wheat and pasta samples prior to HPLC analysis. Samples were extracted with PBS/0.1M EDTA solution and cleaned up through a cartridge containing blank IA-based polymer. The column was washed with PBS (pH 9.2) and the toxin was eluted with methanol and quantified by reversed-phase HPLC with UV detector (lambda=220nm), using methanol:water:acetic acid (15:85:0.1, v/v/v) as the mobile phase. Effective removal of matrix interferences was observed only for pasta with DON recoveries higher than 70% (RSD<7%, n=3) at levels close to or higher than EU regulatory limit.
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Descriptive modelling to predict deoxynivalenol in winter wheat in the Netherlands.
Van Der Fels-Klerx, H J; Burgers, S L G E; Booij, C J H
2010-05-01
Predictions of deoxynivalenol (DON) content in wheat at harvest can be useful for decision-making by stakeholders of the wheat feed and food supply chain. The objective of the current research was to develop quantitative predictive models for DON in mature winter wheat in the Netherlands for two specific groups of end-users. One model was developed for use by farmers in underpinning Fusarium spp. disease management, specifically the application of fungicides around wheat flowering (model A). The second model was developed for industry and food safety authorities, and considered the entire wheat cultivation period (model B). Model development was based on observational data collected from 425 fields throughout the Netherlands between 2001 and 2008. For each field, agronomical information, climatic data and DON levels in mature wheat were collected. Using multiple regression analyses, the set of biological relevant variables that provided the highest statistical performance was selected. The two final models include the following variables: region, wheat resistance level, spraying, flowering date, several climatic variables in the different stages of wheat growing, and length of the period between flowering and harvesting (model B only). The percentages of variance accounted for were 64.4% and 65.6% for models A and B, respectively. Model validation showed high correlation between the predicted and observed DON levels. The two models may be applied by various groups of end-users to reduce DON contamination in wheat-derived feed and food products and, ultimately, reduce animal and consumer health risks.
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Awad, W A; Ghareeb, K; Zentek, J
2014-10-01
Deoxynivalenol (DON), a major contaminant of cereals and grains, is of public health concern worldwide and has been shown to reduce the electrogenic transport of glucose. However, the full effects of Fusarium mycotoxins on nutrient absorption are still not clear. The aim of this study was to investigate whether decreased nutrient absorption was due to specific effects on transporter trafficking in the intestine and whether inhibition of phosphoinositol-3-kinase (PI-3-kinase) affected the electrogenic jejunal transport of glucose. Jejunal mucosa of 6-week-old broiler chickens were mounted in Ussing chambers and treated with DON, wortmannin (a specific inhibitor of PI-3-kinase), DON + wortmannin, phlorizin and cytochalasin B. DON was found to decrease the short-circuit current (Isc) after glucose addition. A similar decline in Isc after glucose addition was observed following pre-application of wortmannin, or phlorizin (Na(+)/glucose co-transporter, SGLT1 inhibitor). The results indicate that DON decreased glucose absorption in the absence of wortmannin or phlorizin but had no additional effect on glucose absorption in their presence. Glucose transport was not affected by cytochalasin B (facilitative glucose transporter, GLUT2 inhibitor). The study provides evidence that the suppressive effect of DON on the electrogenic transport of glucose may be due to an inhibitory activity of the PI3 kinase pathway and intestinal SGLT1. Furthermore, the effect of cytochalasin B on glucose transport in chicken tissues differs from that in mammals. Copyright © 2014 Elsevier Ltd. All rights reserved.
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Ren, Wenkai; Yin, Jie; Hu, Jiayu; Duan, Jielin; Liu, Gang; Tan, Bie; Xiong, Xia; Oso, Abimbola Oladele; Adeola, Olayiwola; Yao, Kang; Yin, Yulong; Li, Tiejun
2014-01-01
Deoxynivalenol (DON) has various toxicological effects in humans and pigs that result from the ingestion of contaminated cereal products. This study was conducted to investigate the protective effects of dietary supplementation with glutamic acid on piglets challenged with DON. A total of 20 piglets weaned at 28 d of age were randomly assigned to receive 1 of 4 treatments (5 piglets/treatment): 1) basal diet, negative control (NC); 2) basal diet +4 mg/kg DON (DON); 3) basal diet +2% (g/g) glutamic acid (GLU); 4) basal diet +4 mg/kg DON +2% glutamic acid (DG). A 7-d adaptation period was followed by 30 days of treatment. A metabolite analysis using nuclear magnetic resonance spectroscopy (1H-NMR)-based metabolomic technology and the determination of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities for plasma, as well as the activity of Caspase-3 and the proliferation of epithelial cells were conducted. The results showed that contents of low-density lipoprotein, alanine, arginine, acetate, glycoprotein, trimethylamine-N-oxide (TMAO), glycine, lactate, and urea, as well as the glutamate/creatinine ratio were higher but high-density lipoprotein, proline, citrate, choline, unsaturated lipids and fumarate were lower in piglets of DON treatment than that of NC treatment (P<0.05). Compared with DON treatment, dietary supplementation with glutamic acid increased the plasma concentrations of proline, citrate, creatinine, unsaturated lipids, and fumarate, and decreased the concentrations of alanine, glycoprotein, TMAO, glycine, and lactate, as well as the glutamate/creatinine ratio (P<0.05). Addition glutamic acid to DON treatment increased the plasma activities of SOD and GSH-Px and the proliferating cell nuclear antigen (PCNA) labeling indexes for the jejunum and ileum (P<0.05). These novel findings indicate that glutamic acid has the potential to repair the injuries associated with oxidative stress as well as the disturbances of energy and amino acid metabolism induced by DON. PMID:25502722
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Wu, Miaomiao; Xiao, Hao; Ren, Wenkai; Yin, Jie; Hu, Jiayu; Duan, Jielin; Liu, Gang; Tan, Bie; Xiong, Xia; Oso, Abimbola Oladele; Adeola, Olayiwola; Yao, Kang; Yin, Yulong; Li, Tiejun
2014-01-01
Deoxynivalenol (DON) has various toxicological effects in humans and pigs that result from the ingestion of contaminated cereal products. This study was conducted to investigate the protective effects of dietary supplementation with glutamic acid on piglets challenged with DON. A total of 20 piglets weaned at 28 d of age were randomly assigned to receive 1 of 4 treatments (5 piglets/treatment): 1) basal diet, negative control (NC); 2) basal diet +4 mg/kg DON (DON); 3) basal diet +2% (g/g) glutamic acid (GLU); 4) basal diet +4 mg/kg DON +2% glutamic acid (DG). A 7-d adaptation period was followed by 30 days of treatment. A metabolite analysis using nuclear magnetic resonance spectroscopy (1H-NMR)-based metabolomic technology and the determination of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities for plasma, as well as the activity of Caspase-3 and the proliferation of epithelial cells were conducted. The results showed that contents of low-density lipoprotein, alanine, arginine, acetate, glycoprotein, trimethylamine-N-oxide (TMAO), glycine, lactate, and urea, as well as the glutamate/creatinine ratio were higher but high-density lipoprotein, proline, citrate, choline, unsaturated lipids and fumarate were lower in piglets of DON treatment than that of NC treatment (P<0.05). Compared with DON treatment, dietary supplementation with glutamic acid increased the plasma concentrations of proline, citrate, creatinine, unsaturated lipids, and fumarate, and decreased the concentrations of alanine, glycoprotein, TMAO, glycine, and lactate, as well as the glutamate/creatinine ratio (P<0.05). Addition glutamic acid to DON treatment increased the plasma activities of SOD and GSH-Px and the proliferating cell nuclear antigen (PCNA) labeling indexes for the jejunum and ileum (P<0.05). These novel findings indicate that glutamic acid has the potential to repair the injuries associated with oxidative stress as well as the disturbances of energy and amino acid metabolism induced by DON.
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Soodoi, Chimidtseren; Sasgary, Soleman; Strasser, Alois; Böhm, Josef
2013-01-01
An experiment was conducted to investigate the individual and combined effects of dietary deoxynivalenol (DON) and a microbial feed additive on plasma cytokine level and on the expression of immune relevant genes in jejunal tissues of broilers. A total of 40 broiler chicks were obtained from a commercial hatchery and divided randomly into four groups (10 birds per group). Birds were reared in battery cages from one day old for 5 weeks. The dietary groups were 1) control birds fed basal diet; 2) DON group fed basal diet contaminated with 10 mg DON/ kg feed; 3) DON + Mycofix group fed basal diet contaminated with 10 mg DON/ kg feed and supplemented with a commercial feed additive, Mycofix® Select (MS) (2.5 kg/ton of feed); 4) Mycofix group fed basal diet supplemented with MS (2.5 kg/ton of feed). At 35 days, the plasma levels of tumor necrosis factor alpha (TNF-α) and interleukin 8 (IL-8) were quantified by ELISA test kits. Furthermore, the mRNA expression of TNF-α, IL-8, IL-1β, interferon gamma (IFNγ), transforming growth factor beta receptor I (TGFBR1) and nuclear factor kappa-light-chain-enhancer of activated B cells 1 (NF-κβ1) in jejunum were quantified by qRT-PCR. The results showed that the plasma TNF-α decreased in response to DON, while in combination with MS, the effect of DON was reduced. DON down-regulated the relative gene expression of IL-1β, TGFBR1 and IFN-γ, and addition of MS to the DON contaminated diet compensates these effects on IL-1β, TGFBR1 but not for IFN-γ. Furthermore, supplementation of MS to either DON contaminated or control diet up-regulated the mRNA expression of NF-κβ1. In conclusion, DON has the potential to provoke and modulate immunological reactions of broilers and subsequently could increase their susceptibility to disease. The additive seemed to have almost as much of an effect as DON, albeit on different genes. PMID:23977054
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Ghareeb, Khaled; Awad, Wageha A; Soodoi, Chimidtseren; Sasgary, Soleman; Strasser, Alois; Böhm, Josef
2013-01-01
An experiment was conducted to investigate the individual and combined effects of dietary deoxynivalenol (DON) and a microbial feed additive on plasma cytokine level and on the expression of immune relevant genes in jejunal tissues of broilers. A total of 40 broiler chicks were obtained from a commercial hatchery and divided randomly into four groups (10 birds per group). Birds were reared in battery cages from one day old for 5 weeks. The dietary groups were 1) control birds fed basal diet; 2) DON group fed basal diet contaminated with 10 mg DON/ kg feed; 3) DON + Mycofix group fed basal diet contaminated with 10 mg DON/ kg feed and supplemented with a commercial feed additive, Mycofix® Select (MS) (2.5 kg/ton of feed); 4) Mycofix group fed basal diet supplemented with MS (2.5 kg/ton of feed). At 35 days, the plasma levels of tumor necrosis factor alpha (TNF-α) and interleukin 8 (IL-8) were quantified by ELISA test kits. Furthermore, the mRNA expression of TNF-α, IL-8, IL-1β, interferon gamma (IFNγ), transforming growth factor beta receptor I (TGFBR1) and nuclear factor kappa-light-chain-enhancer of activated B cells 1 (NF-κβ1) in jejunum were quantified by qRT-PCR. The results showed that the plasma TNF-α decreased in response to DON, while in combination with MS, the effect of DON was reduced. DON down-regulated the relative gene expression of IL-1β, TGFBR1 and IFN-γ, and addition of MS to the DON contaminated diet compensates these effects on IL-1β, TGFBR1 but not for IFN-γ. Furthermore, supplementation of MS to either DON contaminated or control diet up-regulated the mRNA expression of NF-κβ1. In conclusion, DON has the potential to provoke and modulate immunological reactions of broilers and subsequently could increase their susceptibility to disease. The additive seemed to have almost as much of an effect as DON, albeit on different genes.
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Dąbrowski, Michał; Jakimiuk, Ewa; Baranowski, Mirosław; Gajęcka, Magdalena; Zielonka, Łukasz; Gajęcki, Maciej Tadeusz
2017-04-26
Deoxynivalenol (DON) is one of the most prevalent mycotoxins in Europe. Pigs are an animal species that is most susceptible to this mycotoxin. Deoxynivalenol causes significant losses in pig production by lowering feed intake, decreasing daily weight gains, disrupting immune responses, and increasing susceptibility to diseases. The aim of this experiment was to determine the influence of feed contaminated with DON at concentrations insignificantly higher than recommended by the European Commission (900 µg/kg). The experimental feed contained 1008 μg DON/kg. The experiment was performed on eight weaners from the same litter. The animals were randomly divided into two groups: an experimental group (M, n = 4) fed contaminated feed and a control group (C, n = 4) administered feed free of mycotoxins. The experiment lasted for six weeks, and peripheral blood samples were collected from the animals for analyses of selected morphological parameters and changes in the percentages of CD4⁺8 - , CD4 - 8⁺, and CD4⁺8⁺ lymphocytes and antigen-presenting cells (APC) with CD14⁺172⁺ (monocytes), CD172a high 4 - 14 - (conventional dendritic cells, cDC), and CD172a dim 4⁺14 - (plasmacytoid dendritic cells, pDC) phenotypes. The morphological parameters of porcine blood samples were determined by flow cytometry with non-fluorescent particle-size calibration standards, and no differences were observed between groups M and C. An immunophenotyping analysis of lymphocytes and dendritic cells (DC) revealed an increase in the percentage of CD4⁺8 - , CD172a high 4 - 14 - , and CD172a dim 4⁺14 - cells, and a decrease in the number of CD4 - 8⁺ cells in group M. The results of this experiment suggest that prolonged exposure to low doses of DON can change the proportions of immunocompetent cells (a shift towards humoral immunity), without affecting their overall counts.
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Xiao, H; Tan, B E; Wu, M M; Yin, Y L; Li, T J; Yuan, D X; Li, L
2013-10-01
Deoxynivalenol (DON) affects animal and human health and targets the gastrointestinal tract. The objective of this study was to evaluate the ability of composite antimicrobial peptides (CAP) to repair intestinal injury in piglets challenged with DON. A total of 28 piglets (Duroc × Landrace × Large Yorkshire) weaned at 28 d of age were randomly assigned to receive 1 of 4 treatments (7 pigs/treatment): negative control, basal diet (NC), basal diet + 0.4% composite antimicrobial peptide (CAP), basal diet + 4 mg/kg DON (DON), and basal diet + 4 mg/kg DON + 0.4% CAP (DON + CAP). After an adaptation period of 7 d, blood samples were collected on d 15 and 30 after the initiation of treatment for determinations of the concentrations of D-lactate and diamine oxidase. At the end of the study, all piglets were slaughtered to obtain small intestines for the determination of intestinal morphology, epithelial cell proliferation, and protein expression in the mammalian target of rapamycin (mTOR) signaling pathway. The results showed that DON increased serum concentrations of D-lactate and diamine oxidase, and these values in the CAP and DON + CAP treatments were less than those in the NC and DON treatments, respectively (P < 0.05). The villous height/crypt depth in the jejunum and ileum and the goblet cell number in the ileum in the CAP and DON + CAP treatments were greater than those in the NC and DON treatments (P < 0.05). The proliferating cell nuclear antigen (PCNA) labeling indexes for the jejunum and ileum in the DON + CAP treatment were greater than those in the DON treatment (P < 0.05). The DON decreased (P < 0.05) the relative protein expression of phosphorylated Akt (Protein Kinase B) and mTOR in the jejunal and ileal mucosa and of phosphorylated 4E-binding protein 1 (p-4EBP1) in the jejunal mucosa, whereas CAP increased (P < 0.05) the protein expression of p-4EBP1 in the jejunum. These findings showed that DON could enhance intestinal permeability, damage villi, cause epithelial cell apoptosis, and inhibit protein synthesis, whereas CAP improved intestinal morphology and promoted intestinal epithelial cell proliferation and protein synthesis, indicating that CAP may repair the intestinal injury induced by DON.
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Antonissen, G; De Baere, S; Devreese, M; Van Immerseel, F; Martel, A; Croubels, S
2017-01-01
The aim of the present study was to evaluate the effect of the Fusarium mycotoxins deoxynivalenol (DON) and fumonisins (FBs) on the stress response in broiler chickens, using corticosterone (CORT) in plasma as a biomarker. Chickens were fed either a control diet, a DON contaminated diet, a FBs contaminated diet, or a DON and FBs contaminated diet for 15 d at concentrations close to the European Union maximum guidance levels for DON and FBs in poultry. Mean plasma CORT levels were significantly higher in broiler chickens fed a DON contaminated and a DON and FBs contaminated diet compared to birds fed a control diet. A similar trend was observed for animals fed a FBs contaminated diet. Consequently, feeding broilers a diet contaminated with DON and/or FBs induced a CORT stress response, which may indicate a negative effect on animal welfare. © 2016 Poultry Science Association Inc.
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Gunter, Amanda B; Hermans, Anne; Bosnich, Whynn; Johnson, Douglas A; Harris, Linda J; Gleddie, Steve
2016-12-01
Cereal infection by the broad host range fungal pathogen Fusarium graminearum is a significant global agricultural and food safety issue due to the deposition of mycotoxins within infected grains. Methods to study the intracellular effects of mycotoxins often use the baker's yeast model system (Saccharomyces cerevisiae); however, this organism has an efficient drug export network known as the pleiotropic drug resistance (PDR) network, which consists of a family of multidrug exporters. This study describes the first study that has evaluated the potential involvement of all known or putative ATP-binding cassette (ABC) transporters from the PDR network in exporting the F. graminearum trichothecene mycotoxins deoxynivalenol (DON) and 15-acetyl-deoxynivalenol (15A-DON) from living yeast cells. We found that Pdr5p appears to be the only transporter from the PDR network capable of exporting these mycotoxins. We engineered mutants of Pdr5p at two sites previously identified as important in determining substrate specificity and inhibitor susceptibility. These results indicate that it is possible to alter inhibitor insensitivity while maintaining the ability of Pdr5p to export the mycotoxins DON and 15A-DON, which may enable the development of resistance strategies to generate more Fusarium-tolerant crop plants. © 2016 Her Majesty the Queen in Right of Canada. MicrobiologyOpen published by John Wiley & Sons Ltd.
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Schumann, Barbara; Winkler, Janine; Mickenautsch, Nicola; Warnken, Tobias; Dänicke, Sven
2016-08-01
Both deoxynivalenol (DON), zearalenone (ZEN), and their metabolites are known to modulate immune cells in various species whereby viability and proliferation are influenced. Such effects were rarely examined in horses. Therefore, one aim of the present study was to titrate the inhibitory concentrations of DON, 3-acetyl-DON (3AcDON), de-epoxy-DON (DOM-1), ZEN, and α- and β-zearalenol (ZEL) at which viability and proliferation of equine PBMC were reduced by 50 % (IC50) and 10 % (IC10) in vitro. For evaluation of practical relevance of the in vitro findings, a further aim was to screen horses for the background occurrence of DON, ZEN, and their metabolites in systemic circulation and to relate toxin residues both to the inhibitory toxin concentrations and to hematological and clinical-chemical characteristics.The IC50 (μM) for DON, 3AcDON, β-ZEL, α-ZEL, and ZEN were determined at 3.09, 25.90, 75.44, 97.44, and 98.15 in unstimulated cells, respectively, while in proliferating cells, the corresponding IC50 values were 0.73, 6.89, 45.16, 75.96, and 82.51. Neither viability nor proliferation was influenced by DOM-1 up to a concentration of 100 μM.The in vivo screening (N = 49) revealed the occurrence of ZEN (N = 24), α-ZEL (N = 3), β-ZEL (N = 37), DON, and DOM-1 (N = 2). The detected concentrations were much lower than the corresponding IC50 while the IC10 of DON and β-ZEL for proliferating PBMC corresponded to approximately 26 and 35 ng/mL which might be relevant when contaminated diets are fed.Clinical-chemical and hematological traits were not related to mycotoxin residue levels excepting blood urea nitrogen which was positively correlated to the sum of β-ZEL, α-ZEL, and ZEN concentration. Whether this reflects simply the feeding history of the horses or renal failures giving rise to a prolonged half-life of the toxins needs to be clarified further.
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Awad, Wageha A.; Hess, Michael; Twarużek, Magdalena; Grajewski, Jan; Kosicki, Robert; Böhm, Josef; Zentek, Jürgen
2011-01-01
The aim of the present experiment was to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on morphometric indices of jejunum and to follow the passage of deoxynivalenol (DON) through subsequent segments of the digestive tract of broilers. A total of 45 1-d-old broiler chickens (Ross 308 males) were randomly allotted to three dietary treatments (15 birds/treatment): (1) control diet; (2) diet contaminated with 1 mg DON/kg feed; (3) diet contaminated with 5 mg DON/kg feed for five weeks. None of the zootechnical traits (body weight, body weight gain, feed intake, and feed conversion) responded to increased DON levels in the diet. However, DON at both dietary levels (1 mg and 5 mg DON/kg feed) significantly altered the small intestinal morphology. In the jejunum, the villi were significantly (P < 0.01) shorter in both DON treated groups compared with the controls. Furthermore, the dietary inclusion of DON decreased (P < 0.05) the villus surface area in both DON treated groups. The absolute or relative organ weights (liver, heart, proventriculus, gizzard, small intestine, spleen, pancreas, colon, cecum, bursa of Fabricius and thymus) were not altered (P > 0.05) in broilers fed the diet containing DON compared with controls. DON and de-epoxy-DON (DOM-1) were analyzed in serum, bile, liver, feces and digesta from consecutive segments of the digestive tract (gizzard, cecum, and rectum). Concentrations of DON and its metabolite DOM-1 in serum, bile, and liver were lower than the detection limits of the applied liquid chromatography coupled with mass spectrometry (LC-MS/MS) method. Only about 10 to 12% and 6% of the ingested DON was recovered in gizzard and feces, irrespective of the dietary DON-concentration. However, the DON recovery in the cecum as percentage of DON-intake varied between 18 to 22% and was not influenced by dietary DON-concentration. Interestingly, in the present trial, DOM-1 did not appear in the large intestine and in feces. The results indicate that deepoxydation in the present study hardly occurred in the distal segments of the digestive tract, assuming that the complete de-epoxydation occurs in the proximal small intestine where the majority of the parent toxin is absorbed. In conclusion, diets with DON contamination below levels that induce a negative impact on performance could alter small intestinal morphology in broilers. Additionally, the results confirm that the majority of the ingested DON quickly disappears through the gastrointestinal tract. PMID:22174646
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Böswald, C; Engelhardt, G; Vogel, H; Wallnöfer, P R
1995-01-01
The Fusarium mycotoxin zearalenone (ZEA), added at a level of 2 micrograms/ml, was reduced stereoselectively by cultures of Candida tropicalis, Torulaspora delbrückii, Zygosaccharomyces rouxii, and 7 Saccharomyces strains to both alpha- and beta-zearalenol. In contrast, only alpha-zearalenol was produced from ZEA by Pichia fermentans and several yeast strains of the genera Candida, Hansenula, Brettanomyces, Schizosaccharomyces, and Saccharomycopsis. No glucose conjugates of ZEA (zearalenone-4-beta-D-glucopyranoside) were detected. The trichothecene mycotoxin deoxynivalenol (DON) was not metabolized by any of the yeast strains that were used for analysis.
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Bergamini, E; Catellani, D; Dall'asta, C; Galaverna, G; Dossena, A; Marchelli, R; Suman, M
2010-05-01
Fusarium mycotoxins are a relevant problem in the cereal supply chain at a worldwide level, with wheat, maize and barley being the main contaminated crops. Mould growth can happen in the pre-harvest phase and also during transport and storage due to ineffective drying conditions. Among Fusarium toxins, deoxynivalenol (DON) is considered the most important contaminant in wheat due to its widespread occurrence. In the last years the European Food Safety Authority (EFSA) and the European Commission have frequently expressed opinions on Fusarium toxins, setting limits, regulations and guidelines in order to reduce their levels in raw materials and food commodities. In particular, European legislation (Reg. 1881/2006) sets the maximum limit for DON in flour and bread as 750 and 500 microg kg(-1) respectively. Relatively few studies have taken into account the loss of trichothecenes during processing, focusing on how processing factors may influence their degradation. In particular, the description of DON behaviour during bread-making is very difficult, since complex physico-chemical modifications occur during the transformation of the raw ingredients into the final product. In the present study, we studied how DON concentration may be influenced by modifying bread-making parameters, with a special emphasis on the fermentation and baking stages, starting from a naturally contaminated flour at both pilot and industrial scales. Exploiting the power of a Design of Experiments (DoE) approach to consider the great complexity of the studied system, the obtained model shows satisfying goodness-of-fit and prediction, suggesting that the baking step (time/temperature ranges) is crucial for minimizing native DON level in bread.
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Wu, Wenda; Zhou, Hui-Ren; He, Kaiyu; Pan, Xiao; Sugita-Konishi, Yoshiko; Watanabe, Maiko; Zhang, Haibin; Pestka, James J.
2014-01-01
Cereal grain contamination by trichothecene mycotoxins is known to negatively impact human and animal health with adverse effects on food intake and growth being of particular concern. The head blight fungus Fusarium graminearum elaborates five closely related 8-ketotrichothecene congeners: (1) deoxynivalenol (DON), (2) 3-acetyldeoxynivalenol (3-ADON), (3) 15-acetyldeoxynivalenol (15-ADON), (4) fusarenon X (FX), and (5) nivalenol (NIV). While anorexia induction in mice exposed intraperitoneally to DON has been linked to plasma elevation of the satiety hormones cholecystokinin (CCK) and peptide YY3–36 (PYY3–36), the effects of oral gavage of DON or of other 8-keotrichothecenes on release of these gut peptides have not been established. The purpose of this study was to (1) compare the anorectic responses to the aforementioned 8-ketotrichothecenes following oral gavage at a common dose (2.5 mg/kg bw) and (2) relate these effects to changes plasma CCK and PYY3–36 concentrations. Elevation of plasma CCK markedly corresponded to anorexia induction by DON and all other 8-ketotrichothecenes tested. Furthermore, the CCK1 receptor antagonist SR 27897 and the CCK2 receptor antagonist L-365,260 dose-dependently attenuated both CCK- and DON-induced anorexia, which was consistent with this gut satiety hormone being an important mediator of 8-ketotrichothecene-induced food refusal. In contrast to CCK, PYY3–36 was moderately elevated by oral gavage with DON and NIV but not by 3-ADON, 15-ADON, or FX. Taken together, the results suggest that CCK plays a major role in anorexia induction following oral exposure to 8-ketotrichothecenes, whereas PYY3–36 might play a lesser, congener-dependent role in this response. PMID:24385417
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Wu, Wenda; Zhou, Hui-Ren; He, Kaiyu; Pan, Xiao; Sugita-Konishi, Yoshiko; Watanabe, Maiko; Zhang, Haibin; Pestka, James J
2014-04-01
Cereal grain contamination by trichothecene mycotoxins is known to negatively impact human and animal health with adverse effects on food intake and growth being of particular concern. The head blight fungus Fusarium graminearum elaborates five closely related 8-ketotrichothecene congeners: (1) deoxynivalenol (DON), (2) 3-acetyldeoxynivalenol (3-ADON), (3) 15-acetyldeoxynivalenol (15-ADON), (4) fusarenon X (FX), and (5) nivalenol (NIV). While anorexia induction in mice exposed intraperitoneally to DON has been linked to plasma elevation of the satiety hormones cholecystokinin (CCK) and peptide YY₃₋₃₆ (PYY₃₋₃₆), the effects of oral gavage of DON or of other 8-keotrichothecenes on release of these gut peptides have not been established. The purpose of this study was to (1) compare the anorectic responses to the aforementioned 8-ketotrichothecenes following oral gavage at a common dose (2.5 mg/kg bw) and (2) relate these effects to changes plasma CCK and PYY₃₋₃₆ concentrations. Elevation of plasma CCK markedly corresponded to anorexia induction by DON and all other 8-ketotrichothecenes tested. Furthermore, the CCK1 receptor antagonist SR 27897 and the CCK2 receptor antagonist L-365,260 dose-dependently attenuated both CCK- and DON-induced anorexia, which was consistent with this gut satiety hormone being an important mediator of 8-ketotrichothecene-induced food refusal. In contrast to CCK, PYY₃₋₃₆ was moderately elevated by oral gavage with DON and NIV but not by 3-ADON, 15-ADON, or FX. Taken together, the results suggest that CCK plays a major role in anorexia induction following oral exposure to 8-ketotrichothecenes, whereas PYY₃₋₃₆ might play a lesser, congener-dependent role in this response.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Wu, Wenda; Dept. of Food Science and Human Nutrition, Michigan State University, East Lansing, MI 48824; He, Kaiyu
The foodborne mycotoxin deoxynivalenol (DON) induces a ribotoxic stress response in mononuclear phagocytes that mediate aberrant multi-organ upregulation of TNF-α, interleukins and chemokines in experimental animals. While other DON congeners also exist as food contaminants or pharmacologically-active derivatives, it is not known how these compounds affect expression of these cytokine genes in vivo. To address this gap, we compared in mice the acute effects of oral DON exposure to that of seven relevant congeners on splenic expression of representative cytokine mRNAs after 2 and 6 h. Congeners included the 8-ketotrichothecenes 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), fusarenon X (FX), nivalenol (NIV), themore » plant metabolite DON-3-glucoside (D3G) and two synthetic DON derivatives with novel satiety-inducing properties (EN139528 and EN139544). DON markedly induced transient upregulation of TNF-α IL-1β, IL-6, CXCL-2, CCL-2 and CCL-7 mRNA expressions. The two ADONs also evoked mRNA expression of these genes but to a relatively lesser extent. FX induced more persistent responses than the other DON congeners and, compared to DON, was: 1) more potent in inducing IL-1β mRNA, 2) approximately equipotent in the induction of TNF-α and CCL-2 mRNAs, and 3) less potent at upregulating IL-6, CXCL-2, and CCL-2 mRNAs. EN139528's effects were similar to NIV, the least potent 8-ketotrichothecene, while D3G and EN139544 were largely incapable of eliciting cytokine or chemokine mRNA responses. Taken together, the results presented herein provide important new insights into the potential of naturally-occurring and synthetic DON congeners to elicit aberrant mRNA upregulation of cytokines associated with acute and chronic trichothecene toxicity. - Highlights: • We compared effects of DON congeners on biomarker proinflammatory genes in mice. • Oral DON induced splenic IL-1β, IL-6, TNF-α,CXCL-2, CCL-2 and CCL-7 mRNAs. • 8-Ketotrichothecene ranking for biomarkers was FX ≈ DON > 15ADON > 3ADON > NIV. • Plant metabolite DON-3-glucoside failed to induce proinflammatory biomarkers. • Synthetic DON congeners EN139528 and EN139544 did not affect biomarkers.« less
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Pelyhe, Csilla; Kövesi, Benjámin; Zándoki, Erika; Kovács, Balázs; Szabó-Fodor, Judit; Mézes, Miklós; Balogh, Krisztián
2016-12-01
The purpose of this study was to investigate the short-term effects of a single oral dose of T-2 and HT-2 toxin at 0.15, 0.33 and 1.82 mg kg -1 body weight, or deoxynivalenol (DON) and 15-acetyl-DON at 0.13, 0.31 and 1.75 mg kg -1 body weight in common carp. Conjugated dienes and trienes (the early markers of lipid peroxidation) were elevated in all DON-treated groups at the 16th hour, while thiobarbituric acid reactive substances (TBARS; termination marker) were increased at the highest dose of DON at the 16th and 24th hours. T-2 toxin did not cause changes in these parameters. Glutathione content and glutathione peroxidase activity showed higher levels at the 16th hour as the effect of both mycotoxins. The expression of glutathione peroxidase (GPx4) genes (gpx4a and gpx4b) revealed a dual response. Downregulation was observed at the 8th hour, followed by an induction at the 16th hour, at the lowest dose of both mycotoxins. Higher doses revealed long-drawn emergence and an elevation was observed only at the 24th hour. However, at the lowest and highest doses of DON or T-2 toxin the changes in gene expression were delayed, which may be related to the low oxidative stress response, as suggested by the expression profiles of the nrf2, keap1, gpx4a and gpx4b genes.
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Yekkour, Amine; Toumatia, Omrane; Meklat, Atika; Verheecke, Carol; Sabaou, Nasserdine; Zitouni, Abdelghani; Mathieu, Florence
2015-06-01
The cereal-pathogenic Fusarium culmorum (W.G. Smith), causal agent of various blights and rot diseases, is considered as a chronic fungus of economic concern worldwide including North African countries such as Algeria. This pathogen produces a wide range of mycotoxins, amongst which the type B-trichothecene deoxynivalenol (DON). In addition to its acute and chronic side effects in livestock and humans, DON is believed to play a determinant role in the pathogenesis toward Triticeae. However, regardless its significant occurrence and impact, little is known about trichothecenes-producing ability of F. culmorum infecting cereals in Algeria. The PCR assay based on Tri genes of 12 F. culmorum strains (designated Fc1-Fc12), which were recovered from several cropping areas of North Algeria, revealed their trichothecenes-producing ability with 3-AcDON genotype. The molecular prediction was confirmed by HPLC analysis. All strains were able to produce the toxin at detectable levels. Strains Fc1 and Fc12 were the highest producers of this mycotoxin with 220 and 230 µg g(-1), respectively. The evaluation of pathogenic ability of strains through a barley infesting experiment exhibited the significant disease impact of most strains. Significant correlation between the DON-producing ability of strains and the increase in both disease severity (r = 0.88, P = 0.05) and disease occurrence (r = 0.70, P = 0.05) was observed. Chemotyping of F. culmorum isolates and evaluation of their pathogenic ability are reported for the first time for isolates from Algeria, and highlights the important potential of F. culmorum to contaminate cultivated cereal with DON trichothecenes.
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Awad, Wageha A; Ghareeb, Khaled; Bohm, Josef; Zentek, Jurgen
2010-04-01
Trichothecenes are a group of mycotoxins mainly produced by fungi of the Fusarium genus. Deoxynivalenol (DON) is one of the most abundant and important trichothecenes in food and feed, and is a significant contaminants due to its frequent occurrence in toxicologically relevant concentrations worldwide. Since toxin production depends strongly on environmental conditions, such as temperature and humidity, Fusarium toxin contamination can not be avoided completely. Therefore, exposure to this toxin is a permanent health risk for both humans and farm animals. As cereal crops are commonly contaminated with DON and animal diets consist mainly of cereals, it can be assumed that animals are frequently exposed to DON-contaminated feeds. Many strategies can be undertaken to reduce the toxic effect of DON. In addition to the general necessity for minimizing all risk factors that might influence the contamination of cereals with DON, such as the so-called field toxins before harvest, several post-harvest strategies can be applied to counteract possible deleterious effects of this mycotoxin in farm animals. Another approach for decontamination in feedstuffs is the use of adsorbent materials. Adsorbent materials may bind mycotoxins in the gastrointestinal tract and reduce absorption and systemic toxicity. It has been shown that some adsorbents are suitable to alleviate the toxic effects of specific mycotoxins, but its efficacy against trichothecenes is practically zero. Therefore, alternative strategies to reduce animal and human health risk are needed. The use of microbial additives is a method which uses microorganisms having the capability to detoxify mycotoxins by metabolism or degradation prior to their resorption in the gastrointestinal tract. DON has been reported to be completely transformed to de-epoxy-DON by ruminal and intestinal microflora. Eubacterium BBSH 797 was capable of DON degradation and counteracted the toxic effects of DON in animals. This review focuses on the efficacy of microbial feed additives in ameliorating the toxic effects of DON. According to the results of experiments to date, it appears that microorganisms are the main living organisms suitable for this mycotoxin biodegradation. However, the use of this approach depends on its effectiveness from both a practical and economic perspective.
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Wolff, J
2005-12-01
Since national limits have been introduced for the content of DON and ZEA in cereals and cereal products designated for human consumption, it is highly important to understand how these toxins are distributed during sorting, cleaning and further processing to bakery products and pasta. Cereals from several crops were analysed before and after sorting and cleaning. After milling, flours, breads, semolinas, pastas and others were analysed. The results show that that the distribution of DON and ZEA was different. ZEA was more effectively removed than DON. The efficacy of the various processes varied markedly from one lot to the other.
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Using barley genomics to develop Fusarium head blight resistant wheat and barley
USDA-ARS?s Scientific Manuscript database
Fusarium head blight, caused by Fusarium graminearum, is a major problem for wheat and barley growers. During infection, F. graminearum produces trichothecene mycotoxins (e.g., deoxynivalenol or DON) that increases fungal virulence and reduces grain quality and yield. Previous work in Arabidopsis sh...
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USDA-ARS?s Scientific Manuscript database
The Fusarium trichothecenes nivalenol (NIV) and deoxynivalenol (DON) are among the mycotoxins of greatest concern to agricultural production and food/feed safety worldwide. Previous analyses indicate that during early evolution of the Fusarium incarnatum-F. equiseti species complex (FIESC), the tri...
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Meurens, François; Cognie, Juliette; Abrami, Roberta; Oswald, Isabelle P.; Guzylack-Piriou, Laurence
2013-01-01
Background/Aims Deoxynivalenol (DON) is a mycotoxin produced by Fusarium species which is commonly found in temperate regions worldwide as a natural contaminant of cereals. It is of great concern not only in terms of economic losses but also in terms of animal and public health. The digestive tract is the first and main target of this food contaminant and it represents a major site of immune tolerance. A finely tuned cross-talk between the innate and the adaptive immune systems ensures the homeostatic equilibrium between the mucosal immune system and commensal microorganisms. The aim of this study was to analyze the impact of DON on the intestinal immune response. Methodology Non-transformed intestinal porcine epithelial cells IPEC-1 and porcine jejunal explants were used to investigate the effect of DON on the intestinal immune response and the modulation of naive T cells differentiation. Transcriptomic proteomic and flow cytometry analysis were performed. Results DON induced a pro-inflammatory response with a significant increase of expression of mRNA encoding for IL-8, IL-1α and IL-1β, TNF-α in all used models. Additionally, DON significantly induced the expression of genes involved in the differentiation of Th17 cells (STAT3, IL–17A, IL-6, IL-1β) at the expenses of the pathway of regulatory T cells (Treg) (FoxP3, RALDH1). DON also induced genes related to the pathogenic Th17 cells subset such as IL–23A, IL-22 and IL-21 and not genes related to the regulatory Th17 cells (rTh17) such as TGF-β and IL-10. Conclusion DON triggered multiple immune modulatory effects which could be associated with an increased susceptibility to intestinal inflammatory diseases. PMID:23326479
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Weaver, Alexandra C; See, M Todd; Hansen, Jeff A; Kim, Yong B; De Souza, Anna L P; Middleton, Teena F; Kim, Sung Woo
2013-07-17
Three feed additives were tested to improve the growth and health of pigs chronically challenged with aflatoxin (AF) and deoxynivalenol (DON). Gilts (n = 225, 8.8 ± 0.4 kg) were allotted to five treatments: CON (uncontaminated control); MT (contaminated with 150 µg/kg AF and 1100 µg/kg DON); A (MT + a clay additive); B (MT + a clay and dried yeast additive); and C (MT + a clay and yeast culture additive). Average daily gain (ADG) and feed intake (ADFI) were recorded for 42 days, blood collected for immune analysis and tissue samples to measure damage. Feeding mycotoxins tended to decrease ADG and altered the immune system through a tendency to increase monocytes and immunoglobulins. Mycotoxins caused tissue damage in the form of liver bile ductule hyperplasia and karyomegaly. The additives in diets A and B reduced mycotoxin effects on the immune system and the liver and showed some ability to improve growth. The diet C additive played a role in reducing liver damage. Collectively, we conclude that AF and DON can be harmful to the growth and health of pigs consuming mycotoxins chronically. The selected feed additives improved pig health and may play a role in pig growth.
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Weaver, Alexandra C.; See, M. Todd; Hansen, Jeff A.; Kim, Yong B.; De Souza, Anna L. P.; Middleton, Tina F.; Kim, Sung Woo
2013-01-01
Three feed additives were tested to improve the growth and health of pigs chronically challenged with aflatoxin (AF) and deoxynivalenol (DON). Gilts (n = 225, 8.8 ± 0.4 kg) were allotted to five treatments: CON (uncontaminated control); MT (contaminated with 150 µg/kg AF and 1100 µg/kg DON); A (MT + a clay additive); B (MT + a clay and dried yeast additive); and C (MT + a clay and yeast culture additive). Average daily gain (ADG) and feed intake (ADFI) were recorded for 42 days, blood collected for immune analysis and tissue samples to measure damage. Feeding mycotoxins tended to decrease ADG and altered the immune system through a tendency to increase monocytes and immunoglobulins. Mycotoxins caused tissue damage in the form of liver bile ductule hyperplasia and karyomegaly. The additives in diets A and B reduced mycotoxin effects on the immune system and the liver and showed some ability to improve growth. The diet C additive played a role in reducing liver damage. Collectively, we conclude that AF and DON can be harmful to the growth and health of pigs consuming mycotoxins chronically. The selected feed additives improved pig health and may play a role in pig growth. PMID:23867763
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Exposure Assessment for Italian Population Groups to Deoxynivalenol Deriving from Pasta Consumption
Brera, Carlo; Bertazzoni, Valentina; Debegnach, Francesca; Gregori, Emanuela; Prantera, Elisabetta; De Santis, Barbara
2013-01-01
Four hundred and seventy-two pasta samples were collected from long retail distribution chain sales points located in North, Central and South Italy. Representative criteria in the sample collection were followed in terms of number of samples collected, market share, and types of pasta. Samples were analysed by an accredited HPLC-UV method of analysis. The mean contamination level (64.8 μg/kg) of deoxynivalenol (DON) was in the 95th percentile (239 μg/kg) and 99th percentile (337 μg/kg), far below the legal limit (750 μg/kg) set by Regulation EC/1126/2007, accounting for about one tenth, one third and half the legal limit, respectively. Ninety-nine percent of samples fell below half the legal limit. On the basis of the obtained occurrence levels and considering the consumption rates reported by the Italian official database, no health concern was assessed for all consumer groups, being that exposure was far below the Tolerable Daily Intake (TDI) of 1000 ng/kg b.w/day. Nevertheless, despite this, particular attention should be devoted to the exposure to DON by high consumers, such as children aged 3–5 years, who could reach the TDI even with very low levels of DON contamination. PMID:24287568
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Li, FuChang; Wang, JinQuan; Huang, LiBo; Chen, HongJu; Wang, ChunYang
2017-01-01
Deoxynivalenol (DON) is commonly detected in cereals, and is a threat to human and animal health. The effects of microbiological detoxification are now being widely studied. A total of 24 pigs (over four months) were randomly divided into three treatments. Treatment A was fed with a basal diet as the control group. Treatment B was fed with naturally DON-contaminated wheat as a negative control group. Treatment C was fed with a contaminated diet that also had Clostridium sp. WJ06, which was used as a detoxicant. Growth performance, relative organ weight, intestinal morphology, and the intestinal flora of bacteria and fungi were examined. The results showed that after consuming a DON-contaminated diet, the growth performance of the pigs decreased significantly (p < 0.05), the relative organ weight of the liver and kidney increased significantly (p < 0.05), and the integrity of the intestinal barrier was also impaired, though the toxic effects of the contaminated diets on growing pigs were relieved after adding Clostridium sp. WJ06. The data from MiSeq sequencing of the 16S ribosomal ribonucleic acid (rRNA) gene and internal transcribed spacer 1 (ITS1) gene suggested that the abundance of intestinal flora was significantly different across the three treatments. In conclusion, the application of Clostridium sp. WJ06 can reduce the toxic effects of DON and adjust the intestinal microecosystem of growing pigs. PMID:29186895
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Yang, Xin; Li, Long; Duan, Yongle; Yang, Xiaojun
2017-02-01
The aim of this experiment was to study the antioxidant capacity of JM113 isolated from healthy intestinal contents of Tibetan chicken and its protective effect on broiler chickens challenged with deoxynivalenol (DON). Compared with PZ01 and M23, JM113 demonstrated maximum reducing ( < 0.05) activity and resistance in the presence of 1.2 mmol/L hydrogen peroxide, and great scavenging ability ( < 0.05) against hydroxyl, superoxide anion, and 1,1-diphenyl-2-picrylhydrazyl radicals in vitro. For each strain, the antioxidant activities of live bacterial strains were greater ( < 0.05) than of cell free extracts and dead bacterial strains. To examine the antioxidant capacity of JM113 in vivo, 192 1-d-old Arbor Acres chicks were randomly divided into 4 treatments groups consisting of 6 replicates with 8 birds per replicate. The dietary treatments were 1) control; 2) control diet supplemented with JM113 at 1 × 10 cfu/kg; 3) control diet contaminated with DON at 10 mg/kg; 4) control diet contaminated with DON at 10 mg/kg and supplemented with JM113 at 1 × 10 cfu/kg. Dietary supplementation with DON decreased ( < 0.05) superoxide dismutase activity in serum and increased ( < 0.05) malondialdehyde in the jejunal mucosa of broilers, compared to the control. However, supplementation with JM113 to both the DON-contaminated diet and the control diet, caused a significant reduction ( < 0.05) in malondialdehyde activity in the jejunal mucosa. A reduction ( < 0.05) in expression of nuclear factor erythroid 2-related factor 2 was observed in the jejunal mucosa of broilers fed dietary supplementation with DON, whereas the mRNA levels of and its corresponding downstream gene increased ( < 0.05) with JM113 treatment. Addition of JM113 resulted in longer villi ( < 0.05), even in combination with DON compared to the DON group. JM113 treatment, especially in the DON plus JM113 group, up-regulated ( < 0.05) the expression of mRNA. In conclusion, the present study demonstrates that the JM113 strain has great antioxidant activity and supplementation in feed protected the integrity of the intestinal barrier in broilers challenged with DON, suggesting its use for alleviation of negative effects of DON in poultry.
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Mechanisms for ribotoxin-induced ribosomal RNA cleavage
DOE Office of Scientific and Technical Information (OSTI.GOV)
He, Kaiyu; Center for Integrative Toxicology, Michigan State University, East Lansing, MI 48824; Zhou, Hui-Ren
The Type B trichothecene deoxynivalenol (DON), a ribotoxic mycotoxin known to contaminate cereal-based foods, induces ribosomal RNA (rRNA) cleavage in the macrophage via p38-directed activation of caspases. Here we employed the RAW 264.7 murine macrophage model to test the hypothesis that this rRNA cleavage pathway is similarly induced by other ribotoxins. Capillary electrophoresis confirmed that the antibiotic anisomycin (≥ 25 ng/ml), the macrocylic trichothecene satratoxin G (SG) (≥ 10 ng/ml) and ribosome-inactivating protein ricin (≥ 300 ng/ml) induced 18s and 28s rRNA fragmentation patterns identical to that observed for DON. Also, as found for DON, inhibition of p38, double-stranded RNA-activatedmore » kinase (PKR) and hematopoietic cell kinase (Hck) suppressed MAPK anisomycin-induced rRNA cleavage, while, in contrast, their inhibition did not affect SG- and ricin-induced rRNA fragmentation. The p53 inhibitor pifithrin-μ and pan caspase inhibitor Z-VAD-FMK suppressed rRNA cleavage induced by anisomycin, SG and ricin, indicating that these ribotoxins shared with DON a conserved downstream pathway. Activation of caspases 8, 9 and 3 concurrently with apoptosis further suggested that rRNA cleavage occurred in parallel with both extrinsic and intrinsic pathways of programmed cell death. When specific inhibitors of cathepsins L and B (lysosomal cysteine cathepsins active at cytosolic neutral pH) were tested, only the former impaired anisomycin-, SG-, ricin- and DON-induced rRNA cleavage. Taken together, the data suggest that (1) all four ribotoxins induced p53-dependent rRNA cleavage via activation of cathepsin L and caspase 3, and (2) activation of p53 by DON and anisomycin involved p38 whereas SG and ricin activated p53 by an alternative mechanism. Highlights: ► Deoxynivalenol (DON) anisomycin, satratoxin G (SG) and ricin are ribotoxins. ► Ribotoxins induce 18s and 28s rRNA cleavage in the RAW 264.7 macrophage model. ► Ribotoxins induce rRNA cleavage via activation of p53, caspases and cathepsins. ► DON- and anisomycin-triggered rRNA cleavage is p38-dependent. ► SG- and ricin-induced rRNA cleavage is p38-independent.« less
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Bernhoft, Aksel; Høgåsen, Helga R; Rosenlund, Grethe; Ivanova, Lada; Berntssen, Marc H G; Alexander, Jan; Eriksen, Gunnar Sundstøl; Fæste, Christiane Kruse
2017-07-01
Post-smolt Atlantic salmon (Salmo salar) were fed standard feed with added 2 or 6 mg kg -1 pure deoxynivalenol (DON), 0.8 or 2.4 mg kg -1 pure ochratoxin A (OTA), or no added toxins for up to 8 weeks. The experiments were performed in duplicate tanks with 25 fish each per diet group, and the feed was given for three 2-h periods per day. After 3, 6 and 8 weeks, 10 fish from each diet group were sampled. In the following hours after the last feeding at 8 weeks, toxin elimination was studied by sampling three fish per diet group at five time points. Analysis of DON and OTA in fish tissues and plasma was conducted by liquid chromatography-mass spectrometry and high-pressure liquid chromatography with fluorescence detection, respectively. DON was distributed to the liver, kidney, plasma, muscle, skin and brain, and the concentrations in liver and muscle increased significantly from 3 to 8 weeks of exposure to the high-DON diet. After the last feeding at 8 weeks, DON concentration in liver reached a maximum at 1 h and decreased thereafter with a half-life (t 1/2 ) of 6.2 h. DON concentration in muscle reached a maximum at 6 h and was then eliminated with a t 1/2 = 16.5 h. OTA was mainly found in liver and kidney, and the concentration in liver decreased significantly from 3 to 8 weeks in the high-OTA group. OTA was eliminated faster than DON from various tissues. By using Norwegian food consumption data and kinetic findings in this study, we predicted the human exposure to DON and OTA from fish products through carryover from the feed. Following a comparison with tolerable daily intakes, we found the risk to human health from the consumption of salmon-fed diets containing maximum recommended levels of these toxins to be negligible.
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Field Incidence of Mycotoxins in Commercial Popcorn and Potential Environmental Influences
USDA-ARS?s Scientific Manuscript database
Insect ear damage and mycotoxin levels were monitored in several commercial popcorn fields in Central Illinois over a four-year period. Aflatoxin was rare, but fumonisin and deoxynivalenol (DON) were commonly encountered each year and occurred at mean levels in fields up to 1.7 ppm (sample max 2.77...
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USDA-ARS?s Scientific Manuscript database
Fusarium head blight (FHB) or scab is one of the most important plant diseases worldwide, affecting wheat, barley and other small grains. Trichothecene mycotoxins such as deoxynivalenol (DON) accumulate in the grain, presenting a food safety risk and health hazard to humans and animals. Despite cons...
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North American isolates of Fusarium graminearum produce a novel type A trichothecene
USDA-ARS?s Scientific Manuscript database
Deoxynivalenol (DON) is a virulence factor of Fusarium graminearum on wheat and most likely on other host plants. A large scale survey of F. graminearum (sensu strictu) conducted in the northern United States revealed the existence of strains which - based on molecular markers - belong to the 3-acet...
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Production of a new type A trichothecene by isolates of Fusarium graminearum
USDA-ARS?s Scientific Manuscript database
Production of deoxynivalenol (DON) is a virulence factor of Fusarium graminearum on wheat and most likely on other host plants. A large survey of F. graminearum (sensu strictu) in the northern United States revealed the existence of strains which - based on molecular markers - belong to the 3-acetyl...
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USDA-ARS?s Scientific Manuscript database
Using biological control as part of the integrated management of Fusarium head blight (FHB) is understudied yet has considerable potential for significantly contributing to the reduction of FHB and deoxynivalenol (DON) in wheat. However, the development of a cost-effective, commercially feasible li...
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USDA-ARS?s Scientific Manuscript database
Fusarium Head Blight (FHB) is one of the most damaging diseases of wheat. It lowers the grain yield and quality, and contaminates grain with the mycotoxin deoxynivalenol (DON). Genetic resistance is a critical control measure and breeding objective. Many studies have focused on the genetic basis of ...
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Integration of fungicide application and cultivar resistance to manage fusarium head blight in wheat
USDA-ARS?s Scientific Manuscript database
Fusarium head blight (FHB), also known as scab, is a destructive disease of wheat and other small grain cereals. Losses are compounded by the associated mycotoxin deoxynivalenol (DON) which contaminates grain. This chapter provides a brief review of FHB of wheat in North America including occurren...
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USDA-ARS?s Scientific Manuscript database
A rapid, sensitive and multiplexed imaging surface plasmon resonance (iSPR) biosensor assay was developed and validated for three Fusarium toxins, deoxynivalenol (DON), zearalenone (ZEA) and T-2 toxin. The iSPR assay was based on a competitive inhibition format with secondary antibodies (Ab2) conjug...
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Study of Fungal Colonization of Wheat Kernels in Syria with a Focus on Fusarium Species
Alkadri, Dima; Nipoti, Paola; Döll, Katharina; Karlovsky, Petr; Prodi, Antonio; Pisi, Annamaria
2013-01-01
Wheat is one of the main crops in Mediterranean countries, and its cultivation has an important role in the Syrian economy. In Syria, Fusarium head blight (FHB) has not been reported so far. Mycological analysis of 48 samples of wheat kernels collected from cultivation areas with different climatic conditions were performed in 2009 and 2010. Fungal isolates were identified at the genus level morphologically; Fusarium species were characterized morphologically and by species-specific PCR. The most frequent fungal genera found were Alternaria spp. and Cladosporium spp., with frequencies of 24.7% and 8.1%, respectively, while the frequency of Fusarium spp. was 1.5% of kernels. Most frequent Fusarium species were F. tricinctum (30% of all Fusarium isolates), F. culmorum (18%), F. equiseti (14%) and F. graminearum (13%). The mycotoxin production potential of selected Fusarium isolates was assessed by HPLC-MS analysis of rice cultures; chemotyping by PCR was carried out for comparison. All six F. graminearum strains tested produced small amounts (<3 mg/kg) of nivalenol (NIV). All ten F. culmorum strains tested produced large amounts of trichothecenes (>100 mg/kg); four strains produced NIV and six strains produced deoxynivalenol (DON) and 3-acetyl-deoxynivalenol (3Ac-DON). PCR chemotyping lead to an oversimplified picture, because all 3Ac-DON chemotype strains produced more DON than 3Ac-DON; furthermore, the strongest NIV producers produced significant amounts of DON. All tested strains of F. culmorum, F. graminearum, F. pseudograminearum (two strains) and most F. equiseti strains (five of six strains) produced zearalenone. Grains of durum wheat were more frequently colonized by Fusarium spp. than grains of soft wheat. Incidence of Fusarium spp. in irrigated fields was higher than in rainfed fields. The incidence of Fusarium strains producing mycotoxins raises concerns about the risk of Fusarium head blight to Syria and its consequences for public health. PMID:23493058
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NASA Astrophysics Data System (ADS)
Jin, Yu; Liu, Renrong; Zhu, Lixin; Chen, Zhenzhen
2017-11-01
In this paper, an immunochromatographic assay card was developed for the detection of DON in feed and cereals using a novel colloidal gold labeling method. For the colloidal gold immunochromatographic rapid detection (GICD) card, a monoclonal antibody DON-mAb and a goat anti-chicken IgY were drawn on NC membrane as the test line (T line) and the control line (C line) respectively. A gold labeled DON-CBSA conjugate and a gold labeled chicken IgY were sprayed onto the conjugate pad. The GICD card has cut-off levels of 50ng/mL for DON, which is invulnerable to matrix interference, and applicable to a wide range of samples. The GICD detecting results of feed and grain samples were compared with the results of ELISA testing, which showed good consistency.
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Xiao, H; Wu, M M; Tan, B E; Yin, Y L; Li, T J; Xiao, D F; Li, L
2013-10-01
The mycotoxin deoxynivalenol (DON) is a food contaminant that leads to reduced feed intake and reduced BW gain, as well as organ impairment. On the other hand, antimicrobial peptides have been shown to have positive effects on growth performance, nutrient digestibility, and immune function. The purpose of this study was to investigate the protective effects of composite antimicrobial peptides (CAP) on piglets challenged with DON. After a 7-d adaptation period, 28 individually housed piglets (Duroc × Landrace × Large Yorkshire) weaned at 28 d of age were randomly assigned to receive 1 of 4 treatments (7 pigs/treatment): negative control, basal diet (NC), basal diet + 0.4% CAP (CAP), basal diet + 4 mg/kg DON (DON), and basal diet + 4 ppm DON + 0.4% CAP (DON + CAP). On d 15 and 30 after the initiation of treatment, blood samples were collected for the determination of blood profile. Piglets were monitored for 30 d to assess performance and then were slaughtered to obtain organs for the determination of the relative weight of organs. The results showed that dietary supplementation with DON decreased (P < 0.05) ADFI, ADG, and G:F, whereas dietary supplementation with CAP improved ADG and G:F (P < 0.05). The relative weight of the kidney and pancreas was greater and the relative weight of the spleen was lighter in the DON treatment than in the other 3 treatments (P < 0.05). There were no effects (P > 0.05) on other relative weights of viscera, except the relative weight of the gallbladder, but the diamine oxidase activity in the liver decreased in DON-treated piglets (P < 0.05). Piglets in the DON treatment had increased serum concentrations of alkaline phosphatase, alanine transaminase, and aspartate aminotransferase and a dramatic decrease in total protein (P < 0.05), whereas there were no differences (P > 0.05) between the DON + CAP treatment and the other treatments. The DON treatment decreased the numbers of red blood cells and platelets, as well as the serum catalase concentrations, and decreased the serum concentrations of H2O2, maleic dialdehyde, and nitric oxide (P < 0.05). The numbers of platelets and thrombocytocrit, as well as the serum concentrations of catalase, were greater, whereas the maleic dialdehyde concentrations were decreased, in both the CAP and DON + CAP treatments compared with the other treatments (P < 0.05). Compared with the control treatment, DON decreased peripheral lymphocyte proliferation on d 15, whereas supplementation with CAP increased it on d 15 and 30 (P < 0.05). These findings indicate that CAP could improve feed efficiency, immune function, and antioxidation capacity and alleviate organ damage, and thus, it has a protective effect in piglets challenged with DON.
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Ferreira Lopes, Silvia; Vacher, Gaëlle; Savova-Bianchi, Dessislava
2017-01-01
The type B trichothecene mycotoxins deoxynivalenol (DON), nivalenol (NIV) and fusarenon-X (FX) are structurally related secondary metabolites frequently produced by Fusarium on wheat. Consequently, DON, NIV and FX contaminate wheat dusts, exposing grain workers to toxins by inhalation. Those trichothecenes at low, relevant, exposition concentrations have differential effects on intestinal cells, but whether such differences exist with respiratory cells is mostly unknown, while it is required to assess the combined risk of exposure to mycotoxins. The goal of the present study was to compare the effects of DON, NIV and FX alone or in combination on the viability and IL-6 and IL-8-inducing capacity of human epithelial cells representative of the respiratory tract: primary human airway epithelial cells of nasal (hAECN) and bronchial (hAECB) origin, and immortalized human bronchial (16HBE14o-) and alveolar (A549) epithelial cell lines. We report that A549 cells are particularly resistant to the cytotoxic effects of mycotoxins. FX is more toxic than DON and NIV for all epithelial cell types. Nasal and bronchial primary cells are more sensitive than bronchial and alveolar cell lines to combined mycotoxin mixtures at low concentrations, although they are less sensitive to mycotoxins alone. Interactions between mycotoxins at low concentrations are rarely additive and are observed only for DON/NIV and NIV/FX on hAECB cells and DON/NIV/FX on A549 cells. Most interactions at low mycotoxin concentrations are synergistic, antagonistic interactions being observed only for DON/FX on hAECB, DON/NIV on 16HBE14o- and NIV/FX on A549 cells. DON, NIV and FX induce, albeit at different levels, IL-6 and IL-8 release by all cell types. However, NIV and FX at concentrations of low cytotoxicity induce IL-6 release by hAECB and A549 cells, and IL-8 release by hAECN cells. Overall, these data suggest that combined exposure to mycotoxins at low concentrations have a stronger effect on primary nasal epithelial cells than on bronchial epithelial cells and activate different inflammatory pathways. This information is particularly relevant for future studies about the hazard of occupational exposure to mycotoxins by inhalation and its impact on the respiratory tract. PMID:29068378
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Mycotoxin profiling of 1000 beer samples with a special focus on craft beer
van Dam, Ruud; van Doorn, Ronald; Katerere, David; Berthiller, Franz; Haasnoot, Willem; Nielen, Michel W. F.
2017-01-01
Currently beer is booming, mainly due to the steady rise of craft breweries worldwide. Previous surveys for occurrence of mycotoxins in beer, were mainly focussed on industrial produced beer. The present survey reports the presence of mycotoxins in craft beer and how this compares to industrial produced beer. More than 1000 beers were collected from 47 countries, of which 60% were craft beers. A selection of 1000 samples were screened for the presence of aflatoxin B1, ochratoxin A (OTA), zearalenone (ZEN), fumonisins (FBs), T-2 and HT-2 toxins (T-2 and HT-2) and deoxynivalenol (DON) using a mycotoxin 6-plex immunoassay. For confirmatory analysis, a liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and applied. The 6-plex screening showed discrepancies with the LC-MS/MS analysis, possibly due to matrix interference and/or the presence of unknown mycotoxin metabolites. The major mycotoxins detected were DON and its plant metabolite deoxynivalenol-3-β-D-glucopyranoside (D3G). The 6-plex immunoassay reported the sum of DON and D3G (DON+D3G) contaminations ranging from 10 to 475 μg/L in 406 beers, of which 73% were craft beers. The popular craft beer style imperial stout, had the highest percentage of samples suspected positive (83%) with 29% of all imperial stout beers having DON+D3G contaminations above 100 μg/L. LC-MS/MS analysis showed that industrial pale lagers from Italy and Spain, predominantly contained FBs (3–69 μg/L). Besides FBs, African traditional beers also contained aflatoxins (0.1–1.2 μg/L). The presence of OTA, T-2, HT-2, ZEN, β-zearalenol, 3/15-acetyl-DON, nivalenol and the conjugated mycotoxin zearalenone 14-sulfate were confirmed in some beers. This study shows that in 27 craft beers, DON+D3G concentrations occurred above (or at) the Tolerable Daily Intake (TDI). Exceeding the TDI, may have a health impact. A better control of brewing malts for craft beer, should be put in place to circumvent this potential problem. PMID:28982162
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Mycotoxin profiling of 1000 beer samples with a special focus on craft beer.
Peters, Jeroen; van Dam, Ruud; van Doorn, Ronald; Katerere, David; Berthiller, Franz; Haasnoot, Willem; Nielen, Michel W F
2017-01-01
Currently beer is booming, mainly due to the steady rise of craft breweries worldwide. Previous surveys for occurrence of mycotoxins in beer, were mainly focussed on industrial produced beer. The present survey reports the presence of mycotoxins in craft beer and how this compares to industrial produced beer. More than 1000 beers were collected from 47 countries, of which 60% were craft beers. A selection of 1000 samples were screened for the presence of aflatoxin B1, ochratoxin A (OTA), zearalenone (ZEN), fumonisins (FBs), T-2 and HT-2 toxins (T-2 and HT-2) and deoxynivalenol (DON) using a mycotoxin 6-plex immunoassay. For confirmatory analysis, a liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and applied. The 6-plex screening showed discrepancies with the LC-MS/MS analysis, possibly due to matrix interference and/or the presence of unknown mycotoxin metabolites. The major mycotoxins detected were DON and its plant metabolite deoxynivalenol-3-β-D-glucopyranoside (D3G). The 6-plex immunoassay reported the sum of DON and D3G (DON+D3G) contaminations ranging from 10 to 475 μg/L in 406 beers, of which 73% were craft beers. The popular craft beer style imperial stout, had the highest percentage of samples suspected positive (83%) with 29% of all imperial stout beers having DON+D3G contaminations above 100 μg/L. LC-MS/MS analysis showed that industrial pale lagers from Italy and Spain, predominantly contained FBs (3-69 μg/L). Besides FBs, African traditional beers also contained aflatoxins (0.1-1.2 μg/L). The presence of OTA, T-2, HT-2, ZEN, β-zearalenol, 3/15-acetyl-DON, nivalenol and the conjugated mycotoxin zearalenone 14-sulfate were confirmed in some beers. This study shows that in 27 craft beers, DON+D3G concentrations occurred above (or at) the Tolerable Daily Intake (TDI). Exceeding the TDI, may have a health impact. A better control of brewing malts for craft beer, should be put in place to circumvent this potential problem.
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USDA-ARS?s Scientific Manuscript database
Fusarium Head Blight is a worldwide disease of small cereals grains such as wheat. The disease is food safety concern because it produces the metabolite, deoxynivalenol (DON), which is moderately toxic to humans and non-ruminant animals. The current study was implemented to develop more efficient me...
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USDA-ARS?s Scientific Manuscript database
Fusarium graminearum is one of the economically most important plant pathogens causing diseases such as Fusarium Head Blight (FHB) of small grain cereals and ear rot of maize. The mycotoxin deoxynivalenol (DON) produced by F. graminearum is a virulence factor in wheat and probably also on other host...
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USDA-ARS?s Scientific Manuscript database
The inclusion of biological control agent Cryptococcus flavescens OH 182.9 in the integrated management of Fusarium head blight (FHB) has potential for significantly contributing to the reduction of FHB and deoxynivalenol (DON) in wheat. Experiments were conducted to determine if liquid culture gro...
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Mycological survey of Korean cereals and production of mycotoxins by Fusarium isolates.
Lee, U S; Jang, H S; Tanaka, T; Toyasaki, N; Sugiura, Y; Oh, Y J; Cho, C M; Ueno, Y
1986-01-01
The fungal species isolated from Korean cereals (barley, polished barley, wheat, rye, and malt) were Alternaria spp., Aspergillus spp., Chaetomium spp., Drechslera spp., Epicoccum sp., Fusarium spp., and Penicillium spp., etc. The number of Fusarium strains isolated was 36, and their ability to produce Fusarium mycotoxins on rice was tested. Nivalenol (NIV) was produced by Fusarium graminearum (7 of 9 isolates), Fusarium oxysporum (3 of 10 isolates), and Fusarium spp. (7 of 15 isolates). Of 15 isolates of Fusarium spp., 6 formed deoxynivalenol (DON). Fusarenon-X and 3-acetyl-DON were produced by most NIV- and DON-forming isolates, respectively. Zearalenone was produced by 3 isolates of F. graminearum, 1 isolate of Fusarium equiseti, and 11 isolates of Fusarium spp. T-2 toxin was not produced by any Fusarium isolates. The highest concentrations of mycotoxins produced by Fusarium isolates were 77.4 (NIV), 5.3 (DON), 138.3 (fusarenon-X), 40.6 (3-acetyl-DON), and 23.2 (zearalenone) micrograms/g. PMID:2947538
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Mycological survey of Korean cereals and production of mycotoxins by Fusarium isolates.
Lee, U S; Jang, H S; Tanaka, T; Toyasaki, N; Sugiura, Y; Oh, Y J; Cho, C M; Ueno, Y
1986-12-01
The fungal species isolated from Korean cereals (barley, polished barley, wheat, rye, and malt) were Alternaria spp., Aspergillus spp., Chaetomium spp., Drechslera spp., Epicoccum sp., Fusarium spp., and Penicillium spp., etc. The number of Fusarium strains isolated was 36, and their ability to produce Fusarium mycotoxins on rice was tested. Nivalenol (NIV) was produced by Fusarium graminearum (7 of 9 isolates), Fusarium oxysporum (3 of 10 isolates), and Fusarium spp. (7 of 15 isolates). Of 15 isolates of Fusarium spp., 6 formed deoxynivalenol (DON). Fusarenon-X and 3-acetyl-DON were produced by most NIV- and DON-forming isolates, respectively. Zearalenone was produced by 3 isolates of F. graminearum, 1 isolate of Fusarium equiseti, and 11 isolates of Fusarium spp. T-2 toxin was not produced by any Fusarium isolates. The highest concentrations of mycotoxins produced by Fusarium isolates were 77.4 (NIV), 5.3 (DON), 138.3 (fusarenon-X), 40.6 (3-acetyl-DON), and 23.2 (zearalenone) micrograms/g.
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Kolosova, Anna Yu; Sibanda, Liberty; Dumoulin, Frédéric; Lewis, Janet; Duveiller, Etienne; Van Peteghem, Carlos; De Saeger, Sarah
2008-06-02
A lateral-flow immunoassay using a colloidal gold-labelled monoclonal antibody was developed for the rapid detection of deoxynivalenol (DON). Different parameters, such as the amount of immunoreagents, type of the materials, composition of the blocking solution and of the detector reagent mixture, were investigated to provide the optimum assay performance. The experimental results demonstrated that such a visual test had an indicator range rather than a cut-off value. Thus, tests for DON determination with two different indicator ranges of 250-500 and 1000-2000 microg kg(-1) were designed. The method allowed detection of DON at low and high concentration levels, which could be useful for research and practical purposes. The assay applied to spiked wheat and pig feed samples demonstrated accurate and reproducible results. The applicability of the developed lateral-flow test was also confirmed under real field conditions. The test strips prepared in Belgium were sent to Mexico, where they were used for the screening of DON contamination in different bread wheat entries from Fusarium Head Blight inoculated plots. The results were compared with those obtained by ELISA and LC-MS/MS. A poor correlation between ELISA and LC-MS/MS was observed. Visual results of the dipstick tests were in a good agreement with the results of the LC-MS/MS method. Coupled with a simple and fast sample preparation, this qualitative one-step test based on the visual evaluation of results did not require any equipment. Results could be obtained within 10 min. The described assay format can be used as a simple, rapid, cost-effective and robust on-site screening tool for mycotoxin contamination in different agricultural commodities.
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Effect of Prothioconazole Application Timing on Fusarium Mycotoxin Content in Maize Grain.
Limay-Rios, Victor; Schaafsma, Arthur W
2018-05-16
In 2010 and 2011, studies to determine the optimal timing of prothioconazole application (200 g a.i./ha) for reducing Fusarium mycotoxin accumulation in grain were conducted in controlled replicated experiments under small-plot mist-irrigated experiments and in field-scale experiments using two hybrids susceptible to F. gramineaerum infection. A significant decrease in total deoxynivalenol (DON) [DON + 15-acetyl-DON + DON 3-glucoside + 3-acetyl-DON] and zearalenone concentrations was observed when fungicide was sprayed at VT (tasseling) and R1 (silking; P < 0.01) followed by applications at V18 (18th leaf) and R2 (blister; P < 0.05) stages, corresponding to silk completely emerged and fully elongated and to silk emergence and browning, respectively. No reduction in Fusarium graminearum toxins was found after silk senescence (R3 or milk) stage. Moniliformin, fumonisins, beauvericin, enniatins, HT-2 and T-2 toxins were also found in small quantities, and no reduction was observed after treatment ( P > 0.05). Mean reduction (±s.d.) of 59 ± 20% and 57 ± 38% of total DON and zearalenone was observed at full silk elongation, respectively.
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USDA-ARS?s Scientific Manuscript database
Integrated pest management (IPM) is the best available approach for reducing Fusarium head blight (FHB; caused by Fusarium graminearum) and the mycotoxin deoxynivalenol (DON) in wheat grain. Utilizing FHB biological control agent Cryptococcus flavescens OH 182.9 (NRRL Y-30216) as part ...
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Piemontese, Luca; Messia, Maria Cristina; Marconi, Emanuele; Falasca, Luisa; Zivoli, Rosanna; Gambacorta, Lucia; Perrone, Giancarlo; Solfrizzo, Michele
2018-04-01
Deoxynivalenol (DON) is an important mycotoxin produced by several species of Fusarium. It occurs often in wheat grain and is frequently associated with significant levels of its modified form DON-3-glucoside (DON-3-Glc). Ozone (O 3 ) is a powerful disinfectant and oxidant, classified as GRAS (Generally Recognised As Safe), that reacts easily with specific compounds including the mycotoxins aflatoxins, ochratoxin A, trichothecenes and zearalenone. It degrades DON in aqueous solution and can be effective for decontamination of grain. This study reports the efficacy of gaseous ozone treatments in reducing DON, DON-3-Glc, bacteria, fungi and yeasts in naturally contaminated durum wheat. A prototype was used to dispense ozone continuously and homogeneously at different concentrations and exposure time, in 2 kg aliquots of durum wheat. The optimal conditions, which do not affect chemical and rheological parameters of durum wheat, semolina and pasta, were identified (55 g O 3 h -1 for 6 h). The measured mean reductions of DON and DON-3-Glc in ozonated wheat were 29% and 44%, respectively. Ozonation also produced a significant (p < 0.05) reduction of total count (CFU/g) of bacteria, fungi and yeasts in wheat grains.
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Systemic Growth of F. graminearum in Wheat Plants and Related Accumulation of Deoxynivalenol
Moretti, Antonio; Panzarini, Giuseppe; Somma, Stefania; Campagna, Claudio; Ravaglia, Stefano; Logrieco, Antonio F.; Solfrizzo, Michele
2014-01-01
Fusarium head blight (FHB) is an important disease of wheat worldwide caused mainly by Fusarium graminearum (syn. Gibberella zeae). This fungus can be highly aggressive and can produce several mycotoxins such as deoxynivalenol (DON), a well known harmful metabolite for humans, animals, and plants. The fungus can survive overwinter on wheat residues and on the soil, and can usually attack the wheat plant at their point of flowering, being able to infect the heads and to contaminate the kernels at the maturity. Contaminated kernels can be sometimes used as seeds for the cultivation of the following year. Poor knowledge on the ability of the strains of F. graminearum occurring on wheat seeds to be transmitted to the plant and to contribute to the final DON contamination of kernels is available. Therefore, this study had the goals of evaluating: (a) the capability of F. graminearum causing FHB of wheat to be transmitted from the seeds or soil to the kernels at maturity and the progress of the fungus within the plant at different growth stages; (b) the levels of DON contamination in both plant tissues and kernels. The study has been carried out for two years in a climatic chamber. The F. gramineraum strain selected for the inoculation was followed within the plant by using Vegetative Compatibility technique, and quantified by Real-Time PCR. Chemical analyses of DON were carried out by using immunoaffinity cleanup and HPLC/UV/DAD. The study showed that F. graminearum originated from seeds or soil can grow systemically in the plant tissues, with the exception of kernels and heads. There seems to be a barrier that inhibits the colonization of the heads by the fungus. High levels of DON and F. graminearum were found in crowns, stems, and straw, whereas low levels of DON and no detectable levels of F. graminearum were found in both heads and kernels. Finally, in all parts of the plant (heads, crowns, and stems at milk and vitreous ripening stages, and straw at vitreous ripening), also the accumulation of significant quantities of DON-3-glucoside (DON-3G), a product of DON glycosylation, was detected, with decreasing levels in straw, crown, stems and kernels. The presence of DON and DON-3G in heads and kernels without the occurrence of F. graminearum may be explained by their water solubility that could facilitate their translocation from stem to heads and kernels. The presence of DON-3G at levels 23 times higher than DON in the heads at milk stage without the occurrence of F. graminearum may indicate that an active glycosylation of DON also occurs in the head tissues. Finally, the high levels of DON accumulated in straws are worrisome since they represent additional sources of mycotoxin for livestock. PMID:24727554
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Trichothecenes NIV and DON modulate the maturation of murine dendritic cells.
Luongo, D; Severino, L; Bergamo, P; D'Arienzo, R; Rossi, M
2010-01-01
Nivalenol (NIV) and Deoxynivalenol (DON), mycotoxins of the trichothecene family are considered very common food contaminants. In this work, we investigated whether the immunotoxic effects ascribed to these trichothecenes may be mediated by perturbations in the activity of dendritic cells (DCs). Murine bone marrow-derived DCs were used to evaluate the effects of NIV and DON on the LPS-induced maturation process. We found that the expression of the class II MHC and of the accessory CD11c molecules, but not of the costimulatory CD86 marker, was down-regulated by NIV and DON exposure in LPS-treated DCs, as well as nitric oxide (NO) production. Interestingly, NIV, but not DON, induced DC necrosis. Moreover, the analysis of the cytokine pattern showed that IL-12 and IL-10 expressions induced by LPS exposure were suppressed by both trichothecenes in a dose-dependent fashion. On the other hand, the secretion of the proinflammatory cytokine TNF-alpha was increased as a direct consequence of DON and NIV exposure. Taken together, our data indicated that the immunotoxicity of NIV and DON was related to the capacity of both trichothecenes to interfere with phenotypic and functional features of maturing DCs.
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Sayyari, Amin; Fæste, Christiane Kruse; Hansen, Ulrik; Uhlig, Silvio; Framstad, Tore; Schatzmayr, Dian; Sivertsen, Tore
2018-06-01
Deoxynivalenol (DON) is one of the most prevalent Fusarium mycotoxins in grain and can cause economic losses in pig farming due to reduced feed consumption and lower weight gains. Biodetoxification of mycotoxins using bacterial strains has been a focus of research for many years. However, only a few in vivo studies have been conducted on the effectiveness of microbial detoxification of fusariotoxins. This study was therefore aimed at investigating the effect of a feed additive containing the bacterial strain Coriobacteriaceum DSM 11798 (the active ingredient in Biomin® BBSH 797) on growth performance and blood parameters, as well as uptake and metabolism of DON, in growing pigs. Forty-eight crossbred (Landrace-Yorkshire/Duroc-Duroc) weaning pigs were fed with pelleted feed made from naturally contaminated oats, with DON at four concentration levels: (1) control diet (DON < 0.2 mg kg -1 ), (2) low-contaminated diet (DON = 0.92 mg kg -1 ), (3) medium-contaminated diet (DON = 2.2 mg kg -1 ) and (4) high-contaminated diet (DON = 5.0 mg kg -1 ) and equivalent diets containing DSM 11798 as feed additive. During the first 7 days of exposure, pigs in the highest-dose group showed a 20-28% reduction in feed intake and a 24-34% reduction in weight gain compared with pigs in the control and low-dose groups. These differences were levelled out by study completion. Towards the end of the experiment, dose-dependent reductions in serum albumin, globulin and total serum protein were noted in the groups fed with DON-contaminated feed compared with the controls. The addition of DSM 11798 had no effect on the DON-related clinical effects or on the plasma concentrations of DON. The ineffectiveness of the feed additive in the present study could be a consequence of its use in pelleted feed, which might have hindered its rapid release, accessibility or detoxification efficiency in the pig's gastrointestinal tract.
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Role of mycotoxins in herds with and without problems with tail necrosis in neonatal pigs.
Van Limbergen, Tommy; Devreese, Mathias; Croubels, Siska; Broekaert, Nathan; Michiels, Annelies; De Saeger, Sarah; Maes, Dominiek
2017-11-18
This study aimed to investigate a possible involvement of mycotoxins in neonatal tail necrosis in piglets. Ten affected and 10 non-affected farms were selected. Sow feed samples were analysed for the presence of 23 mycotoxins by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Blood plasma samples of sows and their piglets were analysed for the presence of deoxynivalenol (DON), de-epoxydeoxynivalenol, T-2 and HT-2 toxin, zearalenone, alfa-zearalenol, and beta-zearalenol, using LC-MS/MS. Additionally, high-resolution mass spectrometry (HRMS) was performed to detect DON-glucuronide (DON-Glca). There was a significant difference between case herds and control herds for mean DON concentrations in feed and sow plasma. For piglet samples, concentrations of DON were above the limit of quantification of 0.1 ng/ml in only 12 samples. Positive correlations were found between DON concentrations in sow feed and plasma of sows; DON concentration in sow feed and DON-Glca concentration in plasma of sows; and between DON and DON-Glca concentration in sow-plasma. In conclusion, high prevalence of DON in feed samples was found, with significantly higher concentrations in case herds, as well as the presence of DON and DON-Glca in sow plasma. Additional research is needed to identify risk factors, including within-herd factors, associated with neonatal tail necrosis in piglets. © British Veterinary Association (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.
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Phytotoxicity Evaluation of Type B Trichothecenes Using a Chlamydomonas reinhardtii Model System
Suzuki, Tadahiro; Iwahashi, Yumiko
2014-01-01
Type B trichothecenes, which consist of deoxynivalenol (DON) and nivalenol (NIV) as the major end products, are produced by phytotoxic fungi, such as the Fusarium species, and pollute arable fields across the world. The DON toxicity has been investigated using various types of cell systems or animal bioassays. The evaluation of NIV toxicity, however, has been relatively restricted because of its lower level compared with DON. In this study, the Chlamydomonas reinhardtii testing system, which has been reported to have adequate NIV sensitivity, was reinvestigated under different mycotoxin concentrations and light conditions. The best concentration of DON and NIV, and their derivatives, for test conditions was found to be 25 ppm (2.5 × 10−2 mg/mL). In all light test conditions, DON, NIV, and fusarenon-X (FusX) indicated significant growth inhibition regardless of whether a light source existed, or under differential wavelength conditions. FusX growth was also influenced by changes in photon flux density. These results suggest that C. reinhardtii is an appropriate evaluation system for type B trichothecenes. PMID:24476708
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The induction of mycotoxins by trichothecene producing Fusarium species.
Lowe, Rohan; Jubault, Mélanie; Canning, Gail; Urban, Martin; Hammond-Kosack, Kim E
2012-01-01
In recent years, many Fusarium species have emerged which now threaten the productivity and safety of small grain cereal crops worldwide. During floral infection and post-harvest on stored grains the Fusarium hyphae produce various types of harmful mycotoxins which subsequently contaminate food and feed products. This article focuses specifically on the induction and production of the type B sesquiterpenoid trichothecene mycotoxins. Methods are described which permit in liquid culture the small or large scale production and detection of deoxynivalenol (DON) and its various acetylated derivatives. A wheat (Triticum aestivum L.) ear inoculation assay is also explained which allows the direct comparison of mycotoxin production by species, chemotypes and strains with different growth rates and/or disease-causing abilities. Each of these methods is robust and can be used for either detailed time-course studies or end-point analyses. Various analytical methods are available to quantify the levels of DON, 3A-DON and 15A-DON. Some criteria to be considered when making selections between the different analytical methods available are briefly discussed.
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Kłyszejko, Adriana; Kubus, Zaneta; Zakowska, Zofia
2005-01-01
Filamentous fungi are cosmopolitan microorganisms found in almost all environments. It should be pointed out that occurance of moulds on food or feed may cause health disorders in humans and animals. Mycoflora appears as a source of toxic methabolites, mycotoxins, which hepatotoxic, genotoxic, nefrotoxic and carcinogenic abilities were already proven in several studies. Hense mycological analysis of cereal grains raises as an important manner in evaluation of food and feed health features. Among the most frequent cereal contaminants Alternaria, Aspergillus, Fusarium and Penicillium strains are mentioned. Due to their ability to grow on cereals during both its field growth and storage, Fusarium moulds occure to be an important contamination factors in food and feed industry. In this study Fusarium strains isolates from wheat and maize were examined in order to recognize their abilities to produce two toxins: zearalenon (ZEA) and deoxynivalenole (DON). Mycological analysis shown differentiation within fungal microflora occuring in samples of different storage conditions, where Fusarium strains represented aproximately 20-70% of all mould species present. In purpose of Fusarium strains species evaluation, isolates were mycologically analysed. In the second step of the project, toxicological screening of isolates was performed using Thin Liquid Chromatography (TLC) evaluating toxigenic potential of single strains' production of ZEA and DON. This data gives the possibility of pointing the most toxigenic strains and also shows differentiations in their occurance in cereals. This paper presents introductory research data, which can be useful in recognition of cereal contamination with moulds and their toxic methabolites.
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Qiu, Yu-Lou; He, Qing-Hua; Xu, Yang; Wang, Wei; Liu, Yuan-Yuan
2016-01-01
A nanobody (N-28) which can act as a deoxynivalenol (DON) antigen has been generated, and its residues Thr102-Ser106 were identified to bind with anti-DON monoclonal antibody by alanine-scanning mutagenesis. Site-saturation mutagenesis was used to analyze the plasticity of five residues and to improve the sensitivity of the N-28-based immunoassay. After mutagenesis, three mutants were selected by phage immunoassay and were sequenced. The half-maximal inhibitory concentrations of the immunoassay based on mutants N-28-T102Y, N-28-V103L, and N-28-Y105F were 24.49 ± 1.0, 51.83 ± 2.5, and 35.65 ± 1.6 ng/mL, respectively, showing the assay was, respectively, 3.2, 1.5, and 2.2 times more sensitive than the wild-type-based assay. The best mutant, N-28-T102Y, was used to develop a competitive phage ELISA to detect DON in cereals with high specificity and accuracy. In addition, the structural properties of N-28-T102Y and N-28 were investigated, revealing that the affinity of N-28-T102Y decreased because of increased steric hindrance with the large side chain. The lower-binding-affinity antigen mimetic may contribute to the improvement of the sensitivity of competitive immunoassays. These results demonstrate that nanobodies would be a favorable tool for engineering. Moreover, our results have laid a solid foundation for site-saturation mutagenesis of antigen-mimicking nanobodies to improve immunoassay sensitivity for small molecules.
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He, Wei-Jie; Yuan, Qing-Song; Zhang, You-Bing; Guo, Mao-Wei; Gong, An-Dong; Zhang, Jing-Bo; Wu, Ai-Bo; Huang, Tao; Qu, Bo; Li, He-Ping; Liao, Yu-Cai
2016-09-24
Globally, the trichothecene mycotoxins deoxynivalenol (DON) and nivalenol (NIV) are among the most widely distributed mycotoxins that contaminate small grain cereals. In this study, a bacterial consortium, PGC-3, with de-epoxydation activity was isolated from soil by an in situ soil enrichment method. Screening of 14 soil samples that were sprayed with DON revealed that 4 samples were able to biotransform DON into de-epoxydized DON (dE-DON). Among these, the PGC-3 consortium showed the highest and most stable activity to biotransform DON into dE-DON and NIV into dE-NIV. PGC-3 exhibited de-epoxydation activity at a wide range of pH (5-10) and temperatures (20-37 °C) values under aerobic conditions. Sequential subculturing with a continued exposure to DON substantially reduced the microbial population diversity of this consortium. Analyses of the 16S rDNA sequences indicated that PGC-3 comprised 10 bacterial genera. Among these, one species, Desulfitobacterium, showed a steady increase in relative abundance, from 0.03% to 1.55% (a 52-fold increase), as higher concentrations of DON were used in the subculture media, from 0 to 500 μg/mL. This study establishes the foundation to further develop bioactive agents that can detoxify trichothecene mycotoxins in cereals and enables for the characterization of detoxifying genes and their regulation.
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He, Wei-Jie; Yuan, Qing-Song; Zhang, You-Bing; Guo, Mao-Wei; Gong, An-Dong; Zhang, Jing-Bo; Wu, Ai-Bo; Huang, Tao; Qu, Bo; Li, He-Ping; Liao, Yu-Cai
2016-01-01
Globally, the trichothecene mycotoxins deoxynivalenol (DON) and nivalenol (NIV) are among the most widely distributed mycotoxins that contaminate small grain cereals. In this study, a bacterial consortium, PGC-3, with de-epoxydation activity was isolated from soil by an in situ soil enrichment method. Screening of 14 soil samples that were sprayed with DON revealed that 4 samples were able to biotransform DON into de-epoxydized DON (dE-DON). Among these, the PGC-3 consortium showed the highest and most stable activity to biotransform DON into dE-DON and NIV into dE-NIV. PGC-3 exhibited de-epoxydation activity at a wide range of pH (5–10) and temperatures (20–37 °C) values under aerobic conditions. Sequential subculturing with a continued exposure to DON substantially reduced the microbial population diversity of this consortium. Analyses of the 16S rDNA sequences indicated that PGC-3 comprised 10 bacterial genera. Among these, one species, Desulfitobacterium, showed a steady increase in relative abundance, from 0.03% to 1.55% (a 52-fold increase), as higher concentrations of DON were used in the subculture media, from 0 to 500 μg/mL. This study establishes the foundation to further develop bioactive agents that can detoxify trichothecene mycotoxins in cereals and enables for the characterization of detoxifying genes and their regulation. PMID:27669304
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USDA-ARS?s Scientific Manuscript database
The significant and consistent reduction of Fusarium head blight (FHB) and deoxynivalenol (DON) contamination of wheat and barley remains elusive though research results indicate that utilizing an integrated pest management approach achieves the greatest level of disease/toxin control. The use of y...
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Olsson, J; Börjesson, T; Lundstedt, T; Schnürer, J
2002-02-05
Mycotoxin contamination of cereal grains can be detected and quantified using complex extraction procedures and analytical techniques. Normally, the grain odour, i.e. the presence of non-grain volatile metabolites, is used for quality classification of grain. We have investigated the possibility of using fungal volatile metabolites as indicators of mycotoxins in grain. Ten barley samples with normal odour, and 30 with some kind of off-odour were selected from Swedish granaries. The samples were evaluated with regard to moisture content, fungal contamination, ergosterol content, and levels of ochratoxin A (OA) and deoxynivalenol (DON). Volatile compounds were also analysed using both an electronic nose and gas chromatography combined with mass spectrometry (GC-MS). Samples with normal odour had no detectable ochratoxin A and average DON contents of 16 microg kg(-1) (range 0-80), while samples with off-odour had average OA contents of 76 microg kg(-1) (range 0-934) and DON contents of 69 microg kg(-1) (range 0-857). Data were evaluated by multivariate data analysis using projection methods such as principal component analysis (PCA) and partial least squares (PLS). The results show that it was possible to classify the OA level as below or above the maximum limit of 5 microg kg(-1) cereal grain established by the Swedish National Food Administration, and that the DON level could be estimated using PLS. Samples with OA levels below 5 microg kg(-1) had higher concentration of aldehydes (nonanal, 2-hexenal) and alcohols (1-penten-3-ol, 1-octanol). Samples with OA levels above 5 microg kg(-1) had higher concentrations of ketones (2-hexanone, 3-octanone). The GC-MS system predicted OA concentrations with a higher accuracy than the electronic nose, since the GC-MS misclassified only 3 of 37 samples and the electronic nose 7 of 37 samples. No correlation was found between odour and OA level, as samples with pronounced or strong off-odours had OA levels both below and above 5 microg kg(-1). We were able to predict DON levels in the naturally contaminated barley samples using the volatile compounds detected and quantified by either GC-MS or the electronic nose. Pentane, methylpyrazine, 3-pentanone, 3-octene-2-ol and isooctylacetate showed a positive correlation with DON, while ethylhexanol, pentadecane, toluene, 1-octanol, 1-nonanol, and 1-heptanol showed a negative correlation with DON. The root mean square error of estimation values for prediction of DON based on GC-MS and electronic nose data were 16 and 25 microg kg(-1), respectively.
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Grenier, Bertrand; Dohnal, Ilse; Shanmugasundaram, Revathi; Eicher, Susan D.; Selvaraj, Ramesh K.; Schatzmayr, Gerd; Applegate, Todd J.
2016-01-01
Deoxynivalenol (DON) and fumonisins (FB) are the most frequently encountered mycotoxins produced by Fusarium species in livestock diets. The effect of subclinical doses of mycotoxins in chickens is largely unknown, and in particular the susceptibility of birds to pathogenic challenge when fed these fungal metabolites. Therefore, the present study reports the effects of DON and FB on chickens challenged with Eimeria spp, responsible for coccidiosis. Broilers were fed diets from hatch to day 20, containing no mycotoxins, 1.5 mg DON/kg, 20 mg FB/kg, or both toxins (12 pens/diet; 7 birds/pen). At day 14, six pens of birds per diet (half of the birds) were challenged with a 25×-recommended dose of coccidial vaccine, and all birds (challenged and unchallenged) were sampled 6 days later. As expected, performance of birds was strongly affected by the coccidial challenge. Ingestion of mycotoxins did not further affect the growth but repartitioned the rate of reduction (between the fraction due to the change in maintenance and feed efficiency), and reduced apparent nitrogen digestibility. Intestinal lesions and number of oocysts in the jejunal mucosa and feces of challenged birds were more frequent and intense in the birds fed mycotoxins than in birds fed control feed. The upregulation of cytokines (interleukin (IL) IL-1β, IL-6, IL-8 and IL-10) following coccidial infection was higher in the jejunum of birds fed mycotoxins. Further, the higher intestinal immune response was associated with a higher percentage of T lymphocytes CD4+CD25+, also called Tregs, observed in the cecal tonsils of challenged birds fed mycotoxins. Interestingly, the increase in FB biomarker of exposure (sphinganine/sphingosine ratio in serum and liver) suggested a higher absorption and bioavailability of FB in challenged birds. The interaction of DON and FB was very dependent on the endpoint assessed, with three endpoints reporting antagonism, nine additivity, and two synergism. In conclusion, subclinical doses of DON and FB showed little effects in unchallenged chickens, but seem to result in metabolic and immunologic disturbances that amplify the severity of coccidiosis. PMID:27472362
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Grenier, Bertrand; Dohnal, Ilse; Shanmugasundaram, Revathi; Eicher, Susan D; Selvaraj, Ramesh K; Schatzmayr, Gerd; Applegate, Todd J
2016-07-27
Deoxynivalenol (DON) and fumonisins (FB) are the most frequently encountered mycotoxins produced by Fusarium species in livestock diets. The effect of subclinical doses of mycotoxins in chickens is largely unknown, and in particular the susceptibility of birds to pathogenic challenge when fed these fungal metabolites. Therefore, the present study reports the effects of DON and FB on chickens challenged with Eimeria spp, responsible for coccidiosis. Broilers were fed diets from hatch to day 20, containing no mycotoxins, 1.5 mg DON/kg, 20 mg FB/kg, or both toxins (12 pens/diet; 7 birds/pen). At day 14, six pens of birds per diet (half of the birds) were challenged with a 25×-recommended dose of coccidial vaccine, and all birds (challenged and unchallenged) were sampled 6 days later. As expected, performance of birds was strongly affected by the coccidial challenge. Ingestion of mycotoxins did not further affect the growth but repartitioned the rate of reduction (between the fraction due to the change in maintenance and feed efficiency), and reduced apparent nitrogen digestibility. Intestinal lesions and number of oocysts in the jejunal mucosa and feces of challenged birds were more frequent and intense in the birds fed mycotoxins than in birds fed control feed. The upregulation of cytokines (interleukin (IL) IL-1β, IL-6, IL-8 and IL-10) following coccidial infection was higher in the jejunum of birds fed mycotoxins. Further, the higher intestinal immune response was associated with a higher percentage of T lymphocytes CD4⁺CD25⁺, also called Tregs, observed in the cecal tonsils of challenged birds fed mycotoxins. Interestingly, the increase in FB biomarker of exposure (sphinganine/sphingosine ratio in serum and liver) suggested a higher absorption and bioavailability of FB in challenged birds. The interaction of DON and FB was very dependent on the endpoint assessed, with three endpoints reporting antagonism, nine additivity, and two synergism. In conclusion, subclinical doses of DON and FB showed little effects in unchallenged chickens, but seem to result in metabolic and immunologic disturbances that amplify the severity of coccidiosis.
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Effect of food processing on exposure assessment studies with mycotoxins.
Cano-Sancho, German; Sanchis, Vicente; Ramos, Antonio J; Marín, Sonia
2013-01-01
The goals of the present work were, on the one hand, to assess the effect of baking on the stability of zearalenone (ZEA) and deoxynivalenol (DON), as well as the transfer of DON from pasta to boiling water, and, on the other hand, to quantify the impact of DON depletion, during cooking of pasta, on overall exposure estimates. Therefore, the bread-making process was simulated on a pilot-plant scale by using naturally contaminated flour with DON and ZEA. Transfer of DON from pasta to water was evaluated at different boiling times. Pasta was prepared on a pilot-plant scale by using naturally contaminated durum wheat flour; subsequently, it was boiled simulating home cooking. The experiments examined the stability of DON and ZEA during the bread-making process, including fermentation with Saccharomyces cerevisiae and baking at 200°C. Our results showed a high transfer of DON from pasta to boiling water, reaching depletion levels of almost 75%, which correlated with levels found in water. Accordingly, these cooking depletion rates were computed through a stochastic exposure model to weight their impact on the final exposure estimates. Finally, statistically significant differences were found in most of the parameters and populations assessed, but these were not enough to consider the process as protective because the contribution of pasta to the overall DON intake was commonly low.
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Fusarial toxins: secondary metabolites of Fusarium fungi.
Nesic, Ksenija; Ivanovic, Snezana; Nesic, Vladimir
2014-01-01
Exposure to mycotoxins occurs worldwide, even though there are geographic and climatic differences in the amounts produced and occurrence of these substances.Mycotoxins are secondary chemical metabolites of different fungi. They are natural contaminants of cereals, so their presence is often inevitable. Among many genera that produce mycotoxins, Fusarium fungi are the most widespread in cereal-growing areas of the planet. Fusarium fungi produce a diversity of mycotoxin types, whose distributions are also diverse. What is produced and where it is produced is influenced primarily by environmental conditions, and crop production and storage methods. The amount of toxin produced depends on physical (viz., moisture, relative humidity, temperature, and mechanical damage), chemical (viz., carbon dioxide,oxygen, composition of substrate, insecticides and fungicides), and biological factors (viz., plant variety, stress, insects, spore load, etc.). Moisture and temperature have a major influence on mold growth rate and mycotoxin production.Among the most toxic and prevalent fusaria) toxins are the following: zearalenone,fumonisins, moniliformin and trichothecenes (T-2/HT-2 toxin, deoxynivalenol,diacetoxyscirpenol, nivalenol). Zearalenone (ZEA; ZON, F-2 toxin) isaphy to estrogenic compound, primarily a field contaminant, which exhibits estrogenic activity and has been implicated in numerous mycotoxicoses of farm animals,especially pigs. Recently, evidence suggests that ZEA has potential to stimulate the growth of human breast cancer cells. Fumonisins are also cancer-promoting metabolites,of which Fumonisin 8 I (FBI) is the most important. Moniliformin (MON) isalso highly toxic to both animals and humans. Trichothecenes are classified as gastrointestinal toxins, dermatotoxins, immunotoxins, hematotoxins, and gene toxins.T-2 and HT-2 toxin, and diacetoxyscirpenol (DAS, anguidine) are the most toxic mycotoxins among the trichothecene group. Deoxynivalenol (DON, vomitoxin) and nivalenol although less toxic are important because they frequently occur at levels high enough to cause adverse effects.The presence of mycotoxins in the animal diet can produce significant production losses. Any considerable presence of mycotoxins, in major dietary components,confirms the need to adopt a continuous prevention and control program. Such programs are usually based on several common approaches to minimize mycotoxin contamination in the food chain. Major strategies include preventing fungal growth and therefore mycotoxin formation, reducing or eliminating mycotoxins from contaminated feedstuffs, or diverting contaminated products to low risk uses. Because of the complexity of their chemical structures, mycotoxins also present a major analytical challenge. They are also found in a vast array of feed matrices. Analysis is essential for determining the extent of mycotoxin contamination, for risk analysis, confirming the diagnosis of a mycotoxicosis and for monitoring mycotoxin mitigation strategies.For the future, adequately controlling the mycotoxin problem in the livestock economy will depend on implementing appropriate agricultural management policies,as well as augmenting production and storage systems and analysis methods.Only such policies offer the opportunity to bring solid and long-lasting economical results to the livestock industry that is afflicted with the mycotoxin problem.
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Exposure assessment approach through mycotoxin/creatinine ratio evaluation in urine by GC-MS/MS.
Rodríguez-Carrasco, Yelko; Moltó, Juan Carlos; Mañes, Jordi; Berrada, Houda
2014-10-01
In this pilot survey human urine samples were analyzed for presence of 15 mycotoxins and some of their metabolites using a novel urinary multi-mycotoxin GC-MS/MS method following salting-out liquid-liquid extraction. Fifty-four urine samples from children and adults residents in Valencia were analyzed for presence of urinary mycotoxin and expressed in gram of creatinine. Three out of 15 mycotoxins were detected namely, HT-2 toxin, nivalenol and deoxynivalenol (DON). 37 samples showed quantifiable values of mycotoxins. Co-occurrence of these contaminants was also observed in 20.4% of assayed samples. DON was the most frequently detected mycotoxin (68.5%) with mean levels of 23.3 μg/g creatinine (range: 2.8-69.1 μg/g creatinine). The levels of urinary DON were used to carry out an exposure assessment approach. 8.1% of total subjects were estimated to exceed the DON provisional maximum tolerable daily intake (PMTDI) (1 μg/kg b.w.). Two out of 9 exposed children exceeded the DON PMTDI thus, making them the most exposed based on the urinary results. Copyright © 2014 Elsevier Ltd. All rights reserved.
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Feasibility of FT–Raman spectroscopy for rapid screening for DON toxin in ground wheat and barley.
Liu, Y; Delwiche, S R; Dong, Y
2009-10-01
Rapid detection of deoxynivalenol (DON) in cereal-based food and feed has long been the goal of regulators and manufacturers. As non-destructive approaches, infrared (IR) and near-infrared (NIR) spectroscopic techniques have been used for the prediction and classification of contaminated single-kernel and ground grain without any DON extraction steps. These methods, however, are hindered by the intense and broad spectral bands attributed to naturally occurring moisture. Raman spectroscopy could be an alternative to IR and NIR due to its insensitivity to water and fewer overlapped bands. This study explored the feasibility of the Raman technique for rapid and non-destructive screening of DON-contaminated wheat and barley meal. The advantages of this technique include the use of a 1064-nm NIR excitation laser that reduces interference from fluorescence of biological compounds in wheat and barley, the use of a simple intensity-intensity algorithm at two unique frequencies, plus the technique's ease of sample preparation. The results indicate that the simple algorithm, as well as principal component analysis applied to the Raman spectra, can be used to classify low from high DON grain.
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Lohölter, Malte; Meyer, Ulrich; Döll, Susanne; Manderscheid, Remy; Weigel, Hans-Joachim; Erbs, Martin; Höltershinken, Martin; Flachowsky, Gerhard; Dänicke, Sven
2012-11-01
Future livestock production is likely to be affected by both rising ambient temperatures and indirect effects mediated by modified growth conditions of feed plants such as increased atmospheric CO2 concentrations and drought. Corn was grown at elevated CO2 concentrations of 550 ppm and drought stress using free air carbon dioxide enrichment technology. Whole plant silages were generated and fed to sheep kept at three climatic treatments. Differential blood count was performed. Plasma DON and de-epoxy-DON concentration were measured. Warmer environment increased rectal and skin temperatures and respiration rates (p < 0.001 each) but did not affect blood parameters and the almost complete metabolization of DON into de-epoxy-DON. Altered growth conditions of the corn fed did not have single effects on sheep body temperature measures and differential blood count. Though the thermoregulatory activity of sheep was influenced by the thermal environment, the investigated cultivation factors did not indicate considerable impacts on the analysed parameters.
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Frobose, H L; Fruge, E D; Tokach, M D; Hansen, E L; DeRouchey, J M; Dritz, S S; Goodband, R D; Nelssen, J L
2015-03-01
Four experiments were conducted to investigate the effects of deoxynivalenol (DON) from naturally contaminated dried distillers grains with solubles (DDGS) and the efficacy of feed additives in nursery pig diets. In Exp. 1, 180 pigs (10.3 ± 0.2 kg BW) were fed 1 of 5 diets for 21 d. Diets were 1) Positive Control (PC; < 0.5 mg/kg DON), 2) Negative Control (NC; 4 mg/kg DON), 3) NC + 0.10% Biofix (Biomin Inc., Herzogenburg, Austria), 4) NC + 0.15% Cel-can (VAST Inc., Mason City, IA) and 0.50% bentonite clay, and 5) NC + 0.25% Defusion Plus (Cargill Animal Nutrition, Minneapolis, MN). Pigs fed the NC diet had poorer ( < 0.01) ADG than those fed the PC. Pigs fed Defusion Plus had improved ( < 0.03) ADG over those fed NC, whereas pigs fed Biofix or Cel-can with bentonite clay had reduced ADG ( < 0.01) compared with those fed PC. In Exp. 2, 340 pigs (11.7 ± 0.1 kg BW) were fed 1 of 8 diets for 21 d. Diets were 1) PC (< 0.5 mg/kg DON), 2) Low NC (1.5 mg/kg DON), 3) Low NC + 0.15% Biofix, 4) Low NC + 0.30% Biofix, 5) High NC (3.0 mg/kg DON), 6) High NC + 0.30% Biofix, 7) High NC + 0.45% Biofix, and 8) Diet 7 with 5% added water. Increasing the DON level reduced (linear; < 0.05) ADG, ADFI, and pig BW, and Biofix did not improve performance. In Exp. 3, 1,008 pigs (12.5 ± 0.3 kg BW) were fed 6 treatments for 24 d. Diets were 1) PC ( < 0.5 mg/kg DON), 2) NC (3 mg/kg DON), 3) NC + 0.25% Defusion, 4) NC + 0.50% Defusion, 5) Diet 3 with supplemental nutrients, and 6) Diet 5, pelleted. Pigs fed the NC had decreased ( < 0.01) ADG and ADFI, but adding Defusion improved (linear; < 0.04) ADG and ADFI over pigs fed NC. Pelleting improved ( < 0.01) both ADG and G:F, resulting in ADG above PC pigs. In Exp. 4, 980 pigs (12.0 ± 0.3 kg BW) were fed 1 of 7 diets in a 28-d trial in a 2 × 3 + 1 factorial arrangement. The 7 treatments were based on 3 diets fed in meal or pellet form: 1) PC (< 0.5 mg/kg DON), 2) NC (3 mg/kg DON), and 3) NC + 0.25% Defusion. Treatment 7 was Diet 3 with supplemental nutrients in pellet form. No interactions were observed between pelleting and Defusion. Pigs fed the NC had decreased ( < 0.01) ADG and ADFI, and pelleting improved ( < 0.01) ADG to PC levels, driven by improved ( < 0.01) G:F. Adding nutrients or Defusion had no effect. Overall, these studies show that Defusion and pelleting can help overcome some of the negative effects of DON, whereas other feed additives and additional nutrients do not.
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Kosová, Klára; Chrpová, Jana; Šantrůček, Jiří; Hynek, Radovan; Štěrbová, Lenka; Vítámvás, Pavel; Bradová, Jana; Prášil, Ilja Tom
2017-10-03
Fusarium head blight (FHB) disease adversely affects grain quality and final yield in small-grain cereals including barley. In the present study, the effect of an artificial infection with Fusarium culmorum and an application of deoxynivalenol (DON) on barley spikes of cultivars Chevron and Pedant during flowering was investigated at grain mid-dough stage (BBCH 73) 10days after pathogen inoculation (10 dai). Proteomic analysis using a two-dimensional differential gel electrophoresis (2D-DIGE) technique coupled with LC-MS/MS investigated 98 protein spots revealing quantitative or qualitative differences between the experimental variants. Protein functional annotation of 93 identified protein spots revealed that most affected functional groups represent storage proteins (globulins, hordeins), followed by proteins involved in carbohydrate metabolism (α-amylase inhibitor, β-amylase, glycolytic enzymes), amino acid metabolism (aminotransferases), defence response (chitinase, xylanase inhibitor, serpins, SGT1, universal stress protein USP), protein folding (chaperones, chaperonins), redox metabolism (ascorbate-glutathione cycle), and proteasome-dependent protein degradation. The obtained results indicate adverse effects of infection on plant proteome as well as an active plant response to pathogen as shown by enhanced levels of several inhibitors of pathogen-produced degradation enzymes (α-amylase inhibitor, xylanase inhibitor, serpins), chaperones, and other stress-related proteins (SGT1, USP). Genotypic differences were found in hordein abundance between Chevron and Pedant. Copyright © 2017 Elsevier B.V. All rights reserved.
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Zhou, Hongyuan; George, Saji; Hay, Crystal; Lee, Joel; Qian, He; Sun, Xiulan
2017-05-01
To understand the combinatorial toxicity of mycotoxins, we measured the effects of individual, binary and tertiary combinations of Aflatoxin B 1 (AFB 1 ), Deoxynivalenol (DON) and Zearalenone (ZEN) on the cell viability and cellular perturbations of HepG2 and RAW 264.7 cells. The nature of mycotoxins interactions was assessed using mathematical modeling (Chou-Talalay). Mechanisms of cytotoxicity were studied using high content screening (HCS) that probed cytotoxicity responses, such as changes in intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP), intracellular calcium ([Ca 2+ ] i ) flux, and cell membrane damage. Our results showed that individual cytotoxicity of mycotoxins in a decreasing order was DON>AFB 1 >ZEN. Varying combinations of mycotoxins at differing concentrations showed different types of interactions. Most of the mixtures showed increasing toxic effects-synergism and/or addition while antagonistic effects were observed with combination of AFB 1 +ZEN. Generally, combination of mycotoxins showed significantly increased intracellular ROS production and [Ca 2+ ] i flux, and decreased MMP in both cell lines, showing that the synergistic and additive effects of mycotoxin combination originate from perturbations of multiple cellular functions. Additionally, this study demonstrated the applicability of HCS for gaining mechanistic understanding on the toxicity of individual as well as combinatorial mycotoxins, and also provided scientific bases for formulating regulatory policies. Copyright © 2017. Published by Elsevier Ltd.
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Wang, Qinhu; Chen, Daipeng; Wu, Mengchun; Zhu, Jindong; Jiang, Cong; Xu, Jin-Rong; Liu, Huiquan
2018-01-01
Trichothecene mycotoxins, such as deoxynivalenol (DON) produced by the fungal pathogen, Fusarium graminearum, are not only important for plant infection but are also harmful to human and animal health. Trichothecene targets the ribosomal protein Rpl3 that is conserved in eukaryotes. Hence, a self-defense mechanism must exist in DON-producing fungi. It is reported that TRI (trichothecene biosynthesis) 101 and TRI12 are two genes responsible for self-defense against trichothecene toxins in Fusarium. In this study, however, we found that simultaneous disruption of TRI101 and TRI12 has no obvious influence on DON resistance upon exogenous DON treatment in F. graminearum, suggesting that other mechanisms may be involved in self-defense. By using RNA-seq, we identified 253 genes specifically induced in DON-treated cultures compared with samples from cultures treated or untreated with cycloheximide, a commonly used inhibitor of eukaryotic protein synthesis. We found that transporter genes are significantly enriched in this group of DON-induced genes. Of those genes, 15 encode major facilitator superfamily transporters likely involved in mycotoxin efflux. Significantly, we found that genes involved in the metabolism of gamma-aminobutyric acid (GABA), a known inducer of DON production in F. graminearum, are significantly enriched among the DON-induced genes. The GABA biosynthesis gene PROLINE UTILIZATION 2-2 (PUT2-2) is downregulated, while GABA degradation genes are upregulated at least twofold upon treatment with DON, resulting in decreased levels of GABA. Taken together, our results suggest that transporters influencing DON efflux are important for self-defense and that GABA mediates the balance of DON production and self-defense in F. graminearum. PMID:29706976
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Wang, Qinhu; Chen, Daipeng; Wu, Mengchun; Zhu, Jindong; Jiang, Cong; Xu, Jin-Rong; Liu, Huiquan
2018-01-01
Trichothecene mycotoxins, such as deoxynivalenol (DON) produced by the fungal pathogen, Fusarium graminearum , are not only important for plant infection but are also harmful to human and animal health. Trichothecene targets the ribosomal protein Rpl3 that is conserved in eukaryotes. Hence, a self-defense mechanism must exist in DON-producing fungi. It is reported that TRI (trichothecene biosynthesis) 101 and TRI12 are two genes responsible for self-defense against trichothecene toxins in Fusarium . In this study, however, we found that simultaneous disruption of TRI101 and TRI12 has no obvious influence on DON resistance upon exogenous DON treatment in F. graminearum , suggesting that other mechanisms may be involved in self-defense. By using RNA-seq, we identified 253 genes specifically induced in DON-treated cultures compared with samples from cultures treated or untreated with cycloheximide, a commonly used inhibitor of eukaryotic protein synthesis. We found that transporter genes are significantly enriched in this group of DON-induced genes. Of those genes, 15 encode major facilitator superfamily transporters likely involved in mycotoxin efflux. Significantly, we found that genes involved in the metabolism of gamma-aminobutyric acid (GABA), a known inducer of DON production in F. graminearum , are significantly enriched among the DON-induced genes. The GABA biosynthesis gene PROLINE UTILIZATION 2-2 ( PUT2-2 ) is downregulated, while GABA degradation genes are upregulated at least twofold upon treatment with DON, resulting in decreased levels of GABA. Taken together, our results suggest that transporters influencing DON efflux are important for self-defense and that GABA mediates the balance of DON production and self-defense in F. graminearum .
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Khatibi, Piyum A.; McMaster, Nicole J.; Musser, Robert; Schmale, David G.
2014-01-01
Fuel ethanol co-products known as distillers’ dried grains with solubles (DDGS) are a significant source of energy, protein, and phosphorous in animal feed. Fuel ethanol production may concentrate mycotoxins present in corn into DDGS. One hundred and forty one corn DDGS lots collected in 2011 from 78 ethanol plants located in 12 states were screened for the mycotoxins deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-ADON), 3-acetyldeoxynivalenol (3-ADON), nivalenol (NIV), and zearalenone (ZON). DON ranged from <0.50 to 14.62 μg g−1, 15-ADON ranged from <0.10 to 7.55 μg g−1, and ZON ranged from <0.10 to 2.12 μg g−1. None of the DDGS lots contained 3-ADON or NIV. Plants in OH had the highest levels of DON overall (mean of 9.51 μg g−1), and plants in NY, MI, IN, NE, and WI had mean DON levels >1 and <4 μg g−1. Twenty six percent (36/141) of the DDGS lots contained 1.0 to 5.0 μg g−1 DON, 2% (3/141) contained >5.0 and <10.0 μg g−1 DON, and 3% (4/141) contained >10.0 μg g−1 DON. All DDGS lots contaminated with unacceptable levels of DON evaded detection prior to their commercial distribution and were likely sold as feed products. PMID:24674933
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Mastanjević, Kristina; Šarkanj, Bojan; Krska, Rudolf; Sulyok, Michael; Warth, Benedikt; Mastanjević, Krešimir; Šantek, Božidar; Krstanović, Vinko
2018-07-15
The aim was to determine the mycotoxin transfer rate into beer during a semi-industrial production process and the effect of fungicide treatment in the field on mycotoxins concentrations in beer. To ensure the usual practical agronomical conditions, sample A was treated with fungicide Prosaro® 250, and sample B was infected with Fusarium culmorum spores, in order to obtain infected malt. Malt was produced using standard procedure and beer was produced in a semi-industrial unit. During fermentation measurement of sugars (maltotriose and maltose), glycerol and ethanol content was performed on a daily basis. Multiple toxins were determined in malt and beer. Deoxynivalenol (DON), its modified plant metabolite DON-3-glucoside (DON-glucoside), brevianamide F, tryptophol, linamarin, lotaustralin, culmorin (CUL), 15-hydroxy-CUL and 5-hydroyx-CUL were detected in all samples. Results indicate that F. culmorum infection did not influence the fermentation process or the alcohol concentration. Copyright © 2018 Elsevier Ltd. All rights reserved.
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Impact of Climate Change Effects on Contamination of Cereal Grains with Deoxynivalenol
Van der Fels-Klerx, H. J.; van Asselt, Esther D.; Madsen, Marianne S.; Olesen, Jørgen E.
2013-01-01
Climate change is expected to aggravate feed and food safety problems of crops; however, quantitative estimates are scarce. This study aimed to estimate impacts of climate change effects on deoxynivalenol contamination of wheat and maize grown in the Netherlands by 2040. Quantitative modelling was applied, considering both direct effects of changing climate on toxin contamination and indirect effects via shifts in crop phenology. Climate change projections for the IPCC A1B emission scenario were used for the scenario period 2031-2050 relative to the baseline period of 1975-1994. Climatic data from two different global and regional climate model combinations were used. A weather generator was applied for downscaling climate data to local conditions. Crop phenology models and prediction models for DON contamination used, each for winter wheat and grain maize. Results showed that flowering and full maturity of both wheat and maize will advance with future climate. Flowering advanced on average 5 and 11 days for wheat, and 7 and 14 days for maize (two climate model combinations). Full maturity was on average 10 and 17 days earlier for wheat, and 19 and 36 days earlier for maize. On the country level, contamination of wheat with deoxynivalenol decreased slightly, but not significantly. Variability between regions was large, and individual regions showed a significant increase in deoxynivalenol concentrations. For maize, an overall decrease in deoxynivalenol contamination was projected, which was significant for one climate model combination, but not significant for the other one. In general, results disagree with previous reported expectations of increased feed and food safety hazards under climate change. This study illustrated the relevance of using quantitative models to estimate the impacts of climate change effects on food safety, and of considering both direct and indirect effects when assessing climate change impacts on crops and related food safety hazards. PMID:24066059
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Individual and Combined Occurrence of Mycotoxins in Feed Ingredients and Complete Feeds in China.
Ma, Rui; Zhang, Lei; Liu, Meng; Su, Yong-Teng; Xie, Wen-Mei; Zhang, Ni-Ya; Dai, Jie-Fan; Wang, Yun; Rajput, Shahid Ali; Qi, De-Sheng; Karrow, Niel Alexander; Sun, Lv-Hui
2018-03-07
The objective of this study was to investigate the individual and combined contamination of aflatoxin B₁ (AFB₁), zearalenone (ZEN) and deoxynivalenol (DON) in feedstuffs from different Provinces of China between 2016 and 2017. A total of 1569 samples, including 742 feed ingredients and 827 complete pig feed samples, were collected from various regions of China for mycotoxins analysis. The results showed that individual occurrence rates of AFB₁, ZEN, and DON were more than 83.3%, 88%, and 74.5%, respectively, in all the tested samples. DON was the most prevalent contaminant, followed by ZEN and AFB₁, with the average concentrations ranging from 450.0-4381.5 μg/kg, 2.3-729.2 μg/kg, and 1.3-10.0 μg/kg, respectively. Notable, 38.2%, 10.8%, and 0.6% of complete pig feeds were contaminated with DON, ZEN, and AFB₁ over China's regulatory limits, respectively. Moreover, over 75.0% analyzed samples were co-contaminated with two or three mycotoxins. In conclusion, the current study revealed that the feedstuffs in China were severely contaminated with DON, followed by ZEN and AFB₁ during the past two years. These findings highlight the importance of monitoring mycotoxins in livestock feed and implementing feed management and bioremediation strategies to reduce mycotoxin exposure.
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Wu, Li; Wang, Wence; Yao, Kang; Zhou, Ting; Yin, Jie; Li, Tiejun; Yang, Lin; He, Liuqin; Yang, Xiaojian; Zhang, Hongfu; Wang, Qi; Huang, Ruilin; Yin, Yulong
2013-01-01
Deoxynivalenol (DON) is a mycotoxin that reduces feed intake and animal performance, especially in swine. Arginine and glutamine play important roles in swine nutrition. The objective of this study was to determine the effects of dietary supplementation with arginine and glutamine on both the impairment induced by DON stress and immune relevant cytokines in growing pigs. A total of forty 60-d-old healthy growing pigs with a mean body weight of 16.28±1.54 kg were randomly divided into 5 groups, and assigned to 3 amino acid treatments fed 1.0% arginine (Arg), 1.0% glutamine (Gln) and 0.5% Arg+0.5% Gln, respectively, plus a toxin control and a non-toxin control. Pigs in the 3 amino acid treatments were fed the corresponding amino acids, and those in non-toxin control and toxin control were fed commercial diet with 1.64% Alanine as isonitrogenous control for 7 days. The toxin control and amino acid treatments were then challenged by feeding DON-contaminated diet with a final DON concentration of 6 mg/kg of diet for 21 days. No significant differences were observed between toxin control and the amino acid groups with regard to the average daily gain (ADG), although the values for average daily feed intake (ADFI) in the amino acid groups were significantly higher than that in toxin control (P<0.01). The relative liver weight in toxin control was significantly greater than those in non-toxin control, arginine and Arg+Glu groups (P<0.01), but there were no significant differences in other organs. With regard to serum biochemistry, the values of BUN, ALP, ALT and AST in the amino acid groups were lower than those in toxin control. IGF1, GH and SOD in the amino acid groups were significantly higher than those in toxin control (P<0.01). The IL-2 and TNFα values in the amino acid groups were similar to those in non-toxin control, and significantly lower than those in toxin control (P<0.01). These results showed the effects of dietary supplementation with arginine and glutamine on alleviating the impairment induced by DON stress and immune relevant cytokines in growing pigs.
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Joshi, Sweccha; Annida, Rumaisha M; Zuilhof, Han; van Beek, Teris A; Nielen, Michel W F
2016-11-02
A competitive inhibition immunoassay is described for the mycotoxins deoxynivalenol (DON) and ochratoxin A (OTA) in beer using a portable nanostructured imaging surface plasmon resonance (iSPR) biosensor, also referred to as imaging nanoplasmonics. The toxins were directly and covalently immobilized on a 3-dimensional carboxymethylated dextran (CMD) layer on a nanostructured iSPR chip. The assay is based on competition between the immobilized mycotoxins and free mycotoxins in the solution for binding to specific antibodies. The chip surface was regenerated after each cycle, and the combination of CMD and direct immobilization of toxins allowed the chips to be used for more than 450 cycles. The limits of detection (LODs) in beer were 17 ng/mL for DON and 7 ng/mL for OTA (or 0.09 ng/mL after 75 times enrichment). These LODs allowed detection of even less than 10% depletion of the tolerable daily intake of DON and OTA by beer. Significant cross-reactivity of anti-DON was observed toward DON-3-glucoside and 3-acetyl-DON, while no cross-reactivity was seen for 15-acetyl-DON. A preliminary in-house validation with 20 different batches of beer showed that both toxins can be detected at the considered theoretical safe level for beer. The assay can be used for in-field or at-line detection of DON in beer and also in barley without preconcentration, while OTA in beer requires an additional enrichment step, thus making the latter in its present form less suitable for field applications.
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Effect of preceding crop on Fusarium species and mycotoxin contamination of wheat grains.
Qiu, Jianbo; Dong, Fei; Yu, Mingzheng; Xu, Jianhong; Shi, Jianrong
2016-10-01
The Fusarium graminearum species complex infects several cereals and causes the reduction of grain yield and quality. Many factors influence the extent of Fusarium infection and mycotoxin levels. Such factors include crop rotation. In the present study, we explored the effect of rice or maize as former crops on mycotoxin accumulation in wheat grains. More than 97% of samples were contaminated with deoxynivalenol (DON). DON concentrations in wheat grains from rice and maize rotation fields were 884.37 and 235.78 µg kg(-1) . Zearalenone (ZEN) was detected in 45% of samples which were mainly collected from maize-wheat rotation systems. Fusarium strains were isolated and more F. graminearum sensu stricto (s. str.) isolates were cultured from wheat samples obtained from maize rotation fields. DON levels produced by Fusarium isolates from rice rotation fields were higher than those of samples from maize rotation fields. Rice-wheat rotation favours DON accumulation, while more ZEN contamination may occur in maize-wheat rotation models. Appropriate crop rotation may help to reduce toxin levels in wheat grains. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.
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Góral, Tomasz; Wiśniewska, Halina; Ochodzki, Piotr; Walentyn-Góral, Dorota
2016-10-18
Resistance to Fusarium head blight in 32 winter triticale and 34 winter wheat accessions was evaluated. Triticale and wheat were sown in field experiments in two locations. At the time of flowering, heads were inoculated with three Fusarium culmorum isolates. Fusarium head blight index was scored and after the harvest percentage of Fusarium damaged kernels was assessed. Grain was analysed for type B trichothecenes (deoxynivalenol and derivatives, nivalenol) and zearalenone (ZEN) content. The average Fusarium head blight indexes were 28.0% for wheat and 19.2% for triticale accessions. The percentage of Fusarium damaged kernels was also higher for wheat and came to 55.6%, while for triticale this figure was 40.2%. The average content of deoxynivalenol (DON) for wheat amounted to 11.65 mg/kg and was lower than the result for triticale which was 14.12 mg/kg. The average contents of nivalenol were similar in both cereals: 4.13 mg/kg and 5.19 mg/kg for wheat and triticale respectively. Considerable amounts of DON derivatives in the cereals were also detected. The ZEN content in the grain was 0.60 mg/kg for wheat and 0.66 mg/kg for triticale. Relationships between Fusarium head blight index, Fusarium damaged kernels and mycotoxin contents were statistically significant for wheat and mostly insignificant for triticale. Triticale proved to have less infected heads and kernels than wheat. However, the content of type B trichothecenes was higher in triticale grain than in wheat grain.
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Mycotoxin co-occurrence in rice, oat flakes and wheat noodles used as staple foods in Ecuador.
Ortiz, Johana; Van Camp, John; Mestdagh, Frédéric; Donoso, Silvana; De Meulenaer, Bruno
2013-01-01
The co-occurrence of aflatoxin B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂), ochratoxin A (OTA), deoxynivalenol (DON), fumonisin B₁ (FB₁), zearalenone (ZEN), and HT-2 and T-2 toxins in the main Ecuadorian staple cereals (rice, oat flakes, and yellow and white wheat noodles) was evaluated. A ultra high performance liquid chromatography/time-of-flight mass spectrometry (UHPLC/TOFMS) method was developed and validated to screen for the presence of these mycotoxins in those cereal matrices. Matrix-matched calibration curves were used to compensate for ion suppression and extraction losses and the recovery values were in agreement with the minimum requirements of Regulation 401/2006/EC (70-110%). For most mycotoxins, the LODs obtained allowed detection in compliance with the maximum permitted levels set in Regulation EC/2006/1881, with the exception of OTA in all cereals and AFB1 in yellow noodles. Extra target analysis of OTA in oat flakes and wheat noodles was performed by HPLC with fluorescence detection. High rates of contamination were observed in paddy rice (23% DON, 23% FB₁, 7% AFB₁, 2% AFG₁ and 2% AFG₂), white wheat noodles (33% DON and 5% OTA) and oat flakes (17% DON, 2% OTA and 2% AFB₁), whereas the rates of contamination were lower in polished rice (2% AFG₁ and 4% HT-2 toxin) and yellow noodles (5% DON). Low rates of co-occurrence of several mycotoxins were observed only for white wheat noodles (5%) and paddy rice (7%). White noodles were contaminated with DON and/or OTA, while combinations of AFG₁, AFB₁, DON and FB₁ were found in paddy rice. Yellow noodles were contaminated with DON only; oat flakes contained DON, OTA or AFB₁, and polished rice was contaminated with AFG₁ and HT-2 toxin.
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Sobral, M Madalena C; Faria, Miguel A; Cunha, Sara C; Ferreira, Isabel M P L V O
2018-07-01
Aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisin B1 (FB1) and ochratoxin A (OTA) are toxic fungal metabolites co-occurring naturally in the environment. This study aimed to evaluate the toxicological interactions of these mycotoxins concerning additive, antagonistic and synergistic toxicity towards human cells. The theoretical biology-based Combination index-isobologram method was used to evaluate the individual and binary effect of these toxins and determine the type of the interaction using as models Caco-2 (intestinal) and HepG2 (hepatic) cells. Cytotoxicity was assessed using the MTT test at the concentrations of 0.625-20 μM for all the compounds. DON exerted the highest toxicity toward both cells, OTA and AFB1 also showed a dose-effect response, whereas no toxicity was verified for FB1. Synergism or antagonism effects occurred when exposing AFB1-DON and AFB1-OTA on Caco-2 cells at higher or lower concentrations, respectively; while DON-OTA showed synergism throughout all inhibition levels. Concerning HepG2, AFB1-DON exerted a strong synergism, regardless of the level; whereas AFB1-OTA had slight synergism/nearly additive effect; and, OTA-DON had a moderate antagonism/nearly additive effect. Synergistic strengths as high as a dose reduction index of 10 for AFB1-DON were observed in hepatic cells. Taken together our findings indicate that the toxicological effects differ regarding the type of mycotoxins used for combinations and the stronger synergistic effect was observed for mixtures containing DON in both cells. Therefore, even though DON has not been classified as to its carcinogenicity to humans, this mycotoxin may present a serious threat to health, mainly when co-occurring in the environment. Copyright © 2018 Elsevier Ltd. All rights reserved.
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Paper-based immune-affinity arrays for detection of multiple mycotoxins in cereals.
Li, Li; Chen, Hongpu; Lv, Xiaolan; Wang, Min; Jiang, Xizhi; Jiang, Yifei; Wang, Heye; Zhao, Yongfu; Xia, Liru
2018-03-01
Mycotoxins produced by different species of fungi may coexist in cereals and feedstuffs, and could be highly toxic for humans and animals. For quantification of multiple mycotoxins in cereals, we developed a paper-based mycotoxin immune-affinity array. First, paper-based microzone arrays were fabricated by photolithography. Then, monoclonal mycotoxin antibodies were added in a copolymerization reaction with a cross-linker to form an immune-affinity monolith on the paper-based microzone array. With use of a competitive immune-response format, paper-based mycotoxin immune-affinity arrays were successfully applied to detect mycotoxins in samples. The detection limits for deoxynivalenol, zearalenone, T-2 toxin, and HT-2 toxin were 62.7, 10.8, 0.36, and 0.23 μg·kg -1 , respectively, which meet relevant requirements for these compounds in food. The recovery rates were 81-86% for deoxynivalenol, 89-117% for zearalenone, 79-86% for T-2 toxin, and 78-83% for HT-2 toxin, and showed the paper-based immune-affinity arrays had good reproducibility. In summary, the paper-based mycotoxin immune-affinity array provides a sensitive, rapid, accurate, stable, and convenient platform for detection of multiple mycotoxins in agro-foods. Graphical abstract Paper-based immune-affinity monolithic array. DON deoxynivalenol, HT-2 HT-2 toxin, T-2 T-2 toxin, PEGDA polyethylene glycol diacrylate, ZEN zearalenone.
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Nogueira, María Soledad; Decundo, Julieta; Martinez, Mauro; Dieguez, Susana Nelly; Moreyra, Federico; Moreno, Maria Virginia
2018-01-01
Two of the most common species of toxin-producing Fusarium contaminating small cereal grains are Fusarium graminearum and F. poae; with both elaborating diverse toxins, especially deoxynivalenol (DON) and nivalenol (NIV), respectively. The objective of our work during the 2012–2014 growing seasons was to screen crops for the most commonly isolated Fusarium species and to quantify DON and NIV toxins in natural malting-barley samples from different producing areas of Argentina. We identified 1180 Fusarium isolates in the 119 samples analyzed, with 51.2% being F. graminearum, 26.2% F. poae and 22.6% other species. We found high concentrations of mycotoxins, at maximum values of 12 μg/g of DON and 7.71 μg/g of NIV. Of the samples, 23% exhibited DON at an average of 2.36 μg/g, with 44% exceeding the maximum limits (average of 5.24 μg/g); 29% contained NIV at an average of 2.36 μg/g; 7% contained both DON and NIV; and 55% were without DON or NIV. Finally, we report the mycotoxin contamination of the grain samples produced by F. graminearum and F. poae, those being the most frequent Fusarium species present. We identified the main Fusarium species affecting natural malting-barley grains in Argentina and documented the presence of many samples with elevated concentrations of DON and NIV. To our knowledge, the investigation reported here was the first to quantify the contamination by Fusarium and its toxins in natural samples of malting barley in Argentina. PMID:29439459
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Nogueira, María Soledad; Decundo, Julieta; Martinez, Mauro; Dieguez, Susana Nelly; Moreyra, Federico; Moreno, Maria Virginia; Stenglein, Sebastian Alberto
2018-02-11
Two of the most common species of toxin-producing Fusarium contaminating small cereal grains are Fusarium graminearum and F. poae ; with both elaborating diverse toxins, especially deoxynivalenol (DON) and nivalenol (NIV), respectively. The objective of our work during the 2012-2014 growing seasons was to screen crops for the most commonly isolated Fusarium species and to quantify DON and NIV toxins in natural malting-barley samples from different producing areas of Argentina. We identified 1180 Fusarium isolates in the 119 samples analyzed, with 51.2% being F. graminearum , 26.2% F. poae and 22.6% other species. We found high concentrations of mycotoxins, at maximum values of 12 μg/g of DON and 7.71 μg/g of NIV. Of the samples, 23% exhibited DON at an average of 2.36 μg/g, with 44% exceeding the maximum limits (average of 5.24 μg/g); 29% contained NIV at an average of 2.36 μg/g; 7% contained both DON and NIV; and 55% were without DON or NIV. Finally, we report the mycotoxin contamination of the grain samples produced by F. graminearum and F. poae , those being the most frequent Fusarium species present. We identified the main Fusarium species affecting natural malting-barley grains in Argentina and documented the presence of many samples with elevated concentrations of DON and NIV. To our knowledge, the investigation reported here was the first to quantify the contamination by Fusarium and its toxins in natural samples of malting barley in Argentina.
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Field incidence of mycotoxins in commercial popcorn and potential environmental influences.
Dowd, Patrick F; Johnson, Eric T
2010-02-01
Popcorn ear damage by insects and mycotoxin levels in kernels were monitored in several commercial popcorn fields in central Illinois over a 4-year period. Aflatoxin was rare, but fumonisin and deoxynivalenol (DON) were commonly encountered each year, and occurred at mean levels in fields up to 1.7 mg/kg (sample max. 2.77 mg/kg) and 1.9 mg/kg (sample max. 2.66 mg/kg), respectively. Neither fumonisin nor DON levels were significantly correlated with the percent of ears with visibly moldy insect-damaged kernels. Significant correlations were noted for the percent of ears with early caterpillar damage and both fumonisin and DON levels overall for some years and at specific sites in other years. Fumonisin levels were generally more highly correlated with insect damage than DON levels. Insect damaged kernels had 100- to 500-fold or greater levels of fumonisin compared to noninsect-damaged kernels, while DON levels were closer to 10- to 30-fold higher in insect damaged versus nondamaged kernels. A high percentage of DON-contaminated kernels were not insect damaged in 2007 and 2008. In some cases, differing mycotoxin levels for the same hybrid and same year planted at different locations appeared to be due to the prior crop. Higher DON levels in 2008 than other years were most likely associated with higher levels of rainfall and cooler temperatures than average during ear fill. While kernel sorters are reported to remove mycotoxin-contaminated popcorn kernels to acceptible levels, consideration of environmental factors that promote mycotoxins in popcorn should result in more effective control measures in the field.
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Chronic DON exposure and acute LPS challenge: effects on porcine liver morphology and function.
Renner, Lydia; Kahlert, Stefan; Tesch, Tanja; Bannert, Erik; Frahm, Jana; Barta-Böszörményi, Anikó; Kluess, Jeannette; Kersten, Susanne; Schönfeld, Peter; Rothkötter, Hermann-Josef; Dänicke, Sven
2017-08-01
The aim of the present study was to examine the role of chronic deoxynivalenol (DON) exposition on the liver morphology and function in combination with pre- and post-hepatic lipopolysaccharide (LPS) stress in young pigs fed for 4 weeks with a DON-contaminated diet (4.59 mg/kg feed). At the end of the experiment, LPS (7.5 μg/kg BW) was administered for 1 h pre-hepatically (Vena portae hepatis) or post-hepatically (Vena jugularis). Liver morphology was macroscopically checked and showed haemorrhage in all LPS groups, significantly higher relative liver weights, accompanied by marked oedema in the gallbladder wall. Histological changes were judged by a modified histology activity index (HAI). Liver HAI score was significantly increased in all LPS groups compared to placebo, primarily due to neutrophil infiltration and haemorrhage. DON feed alone was without effect on the liver HAI. Liver function was characterized by (i) hepatic biochemical markers, (ii) mitochondrial respiration and (iii) Ca 2+ accumulation capacity of isolated mitochondria. Clinical chemical parameters characterizing liver function were initially (<3 h) slightly influenced by LPS. After 3 h, bilirubin and alkaline phosphatase were increased significantly, in DON-fed, jugular-infused LPS group. Respiration and Ca 2+ accumulation capacity of isolated liver mitochondria was not impaired by chronic DON exposure, acute LPS challenge or combined treatments. DON-contaminated feed did not change macroscopy and histology of the liver, but modified the function under LPS stress. The different function was not linked to modifications of liver mitochondria.
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Kinoshita, Asako; Keese, Christina; Meyer, Ulrich; Starke, Alexander; Wrenzycki, Christine; Dänicke, Sven
2018-01-01
The objective of this study was to investigate the effect of long-term exposure to a Fusarium toxin deoxynivalenol (DON, 5 mg/kg DM) on the energy metabolism in lactating cows fed diets with different amounts of concentrate. In Period 1 27 German Holstein cows were assigned to two groups and fed a control or mycotoxin-contaminated diet with 50% concentrate for 11 weeks. In Period 2 each group was further divided and fed either a diet containing 30% or 60% concentrate for 16 weeks. Blood samples were collected in week 0, 4, 8, 15, 21, and 27 for calculation of the Revised Quantitative Insulin Sensitivity Check Index and biopsy samples of skeletal muscle and the liver in w 0, 15, and 27 for analysis by real-time RT-qPCR. The DON-fed groups presented lower insulin sensitivities than controls at week 27. Concomitantly, muscular mRNA expression of insulin receptors and hepatic mRNA expression of glucose transporter 2 and key enzymes for gluconeogenesis and fatty acid metabolism were lower in DON-fed cows compared to the control. The study revealed no consistent evidence that DON effects were modified by dietary concentrate levels. In conclusion, long-term dietary DON intake appears to have mild effects on energy metabolism in lactating dairy cows. PMID:29738450
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Individual and Combined Occurrence of Mycotoxins in Feed Ingredients and Complete Feeds in China
Ma, Rui; Zhang, Lei; Liu, Meng; Su, Yong-Teng; Xie, Wen-Mei; Zhang, Ni-Ya; Dai, Jie-Fan; Wang, Yun; Qi, De-Sheng; Karrow, Niel Alexander; Sun, Lv-Hui
2018-01-01
The objective of this study was to investigate the individual and combined contamination of aflatoxin B1 (AFB1), zearalenone (ZEN) and deoxynivalenol (DON) in feedstuffs from different Provinces of China between 2016 and 2017. A total of 1569 samples, including 742 feed ingredients and 827 complete pig feed samples, were collected from various regions of China for mycotoxins analysis. The results showed that individual occurrence rates of AFB1, ZEN, and DON were more than 83.3%, 88%, and 74.5%, respectively, in all the tested samples. DON was the most prevalent contaminant, followed by ZEN and AFB1, with the average concentrations ranging from 450.0–4381.5 μg/kg, 2.3–729.2 μg/kg, and 1.3–10.0 μg/kg, respectively. Notable, 38.2%, 10.8%, and 0.6% of complete pig feeds were contaminated with DON, ZEN, and AFB1 over China’s regulatory limits, respectively. Moreover, over 75.0% analyzed samples were co-contaminated with two or three mycotoxins. In conclusion, the current study revealed that the feedstuffs in China were severely contaminated with DON, followed by ZEN and AFB1 during the past two years. These findings highlight the importance of monitoring mycotoxins in livestock feed and implementing feed management and bioremediation strategies to reduce mycotoxin exposure. PMID:29518909
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Edwards, S. G.; Pirgozliev, S. R.; Hare, M. C.; Jenkinson, P.
2001-01-01
We developed a PCR-based assay to quantify trichothecene-producing Fusarium based on primers derived from the trichodiene synthase gene (Tri5). The primers were tested against a range of fusarium head blight (FHB) (also known as scab) pathogens and found to amplify specifically a 260-bp product from 25 isolates belonging to six trichothecene-producing Fusarium species. Amounts of the trichothecene-producing Fusarium and the trichothecene mycotoxin deoxynivalenol (DON) in harvested grain from a field trial designed to test the efficacies of the fungicides metconazole, azoxystrobin, and tebuconazole to control FHB were quantified. No correlation was found between FHB severity and DON in harvested grain, but a good correlation existed between the amount of trichothecene-producing Fusarium and DON present within grain. Azoxystrobin did not affect levels of trichothecene-producing Fusarium compared with those of untreated controls. Metconazole and tebuconazole significantly reduced the amount of trichothecene-producing Fusarium in harvested grain. We hypothesize that the fungicides affected the relationship between FHB severity and the amount of DON in harvested grain by altering the proportion of trichothecene-producing Fusarium within the FHB disease complex and not by altering the rate of DON production. The Tri5 quantitative PCR assay will aid research directed towards reducing amounts of trichothecene mycotoxins in food and animal feed. PMID:11282607
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Savi, Geovana D; Piacentini, Karim C; de Souza, Stephany Ramos; Costa, Maíra E B; Santos, Cristina M R; Scussel, Vildes M
2015-07-16
The efficiency of zinc compounds (zinc sulfate, ZnSO4 and zinc oxide, ZnO in regular and nanosize, respectively) on wheat plants was evaluated against growth of Fusarium graminearum and DON formation. In addition, any possible effects on the grain microstructures were observed by scanning electron microscopy (SEM), and the remaining residue of Zn on wheat plants was analyzed. The plants were inoculated with F. graminearum and treated with Zn compounds (100mM) onto spikelets at the anthesis stage. When wheat plants reached maturation, grains were harvested and evaluated for Fusarium (number of colonies, CFU/g), DON formation, and SEM observation, followed by determination of possible remaining Zn residue. The groups treated with ZnSO4 and ZnO-NP showed a reduction in number of CFU of F. graminearum when compared to the control. Similarly for DON formation, i.e. the toxin was reduced to non-detected levels in the treated group. ZnO-NP efficiently reduced F. graminearum and DON formation in the grains at low concentration. Zn remained within the international recommended level for consumption and the treatment did not cause any damage to wheat grains. New strategies of control using Zn compounds in addition to conventional treatments could increase the efficiency against FBH and DON formation. Copyright © 2015 Elsevier B.V. All rights reserved.
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Winter, Mark; Koopmann, Birger; Döll, Katharina; Karlovsky, Petr; Kropf, Ute; Schlüter, Klaus; von Tiedemann, Andreas
2013-07-01
Factors limiting trichothecene contamination of mature wheat grains after Fusarium infection are of major interest in crop production. In addition to ear infection, systemic translocation of deoxynivalenol (DON) may contribute to mycotoxin levels in grains after stem base infection with toxigenic Fusarium spp. However, the exact and potential mechanisms regulating DON translocation into wheat grains from the plant base are still unknown. We analyzed two wheat cultivars differing in susceptibility to Fusarium head blight (FHB), which were infected at the stem base with Fusarium culmorum in climate chamber experiments. Fungal DNA was found only in the infected stem base tissue, whereas DON and its derivative, DON-3-glucoside (D3G), were detected in upper plant parts. Although infected stem bases contained more than 10,000 μg kg⁻¹ dry weight (DW) of DON and mean levels of DON after translocation in the ear and husks reached 1,900 μg kg⁻¹ DW, no DON or D3G was detectable in mature grains. D3G quantification revealed that DON detoxification took mainly place in the stem basis, where ≤ 50% of DON was metabolized into D3G. Enhanced expression of a gene putatively encoding a uridine diphosphate-glycosyltransferase (GenBank accession number FG985273) was observed in the stem base after infection with F. culmorum. Resistance to F. culmorum stem base infection, DON glycosylation in the stem base, and mycotoxin translocation were unrelated to cultivar resistance to FHB. Histological studies demonstrated that the vascular transport of DON labeled with fluorescein as a tracer from the peduncle to the grain was interrupted by a barrier zone at the interface between grain and rachilla, formerly described as "xylem discontinuity". This is the first study to demonstrate the effective control of influx of systemically translocated fungal mycotoxins into grains at the rachilla-seed interface by the xylem discontinuity tissue in wheat ears.
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Mitterbauer, R; Heinrich, M; Rauscher, R; Lemmens, M; Bürstmayr, H; Adam, G
2003-03-01
Infection withFusarium graminearum andF. culmorum not only causes severe yield and quality losses, the most relevant concern is the contamination of cereal foods and feeds with trichothecenes (e.g. deoxynivalenol, DON). The ability to synthesize trichothecenes has been shown to be a virulence factor ofF. graminearum on wheat and, on the other hand, toxin resistance is most likely an important component of field resistance. Our hypothesis is that pleiotropic drug resistance mediated by PDR-type ABC transporter proteins (acting as membrane located drug efflux pumps) is a relevant mechanism of DON resistance not only in yeast but also in wheat. Goal of this project is the development of molecular markers for this gene family for use in marker-assisted plant breeding programs. The technical difficulties caused by the large size of the PDR-family are discussed.
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Góral, Tomasz; Wiśniewska, Halina; Ochodzki, Piotr; Walentyn-Góral, Dorota
2016-01-01
Resistance to Fusarium head blight in 32 winter triticale and 34 winter wheat accessions was evaluated. Triticale and wheat were sown in field experiments in two locations. At the time of flowering, heads were inoculated with three Fusarium culmorum isolates. Fusarium head blight index was scored and after the harvest percentage of Fusarium damaged kernels was assessed. Grain was analysed for type B trichothecenes (deoxynivalenol and derivatives, nivalenol) and zearalenone (ZEN) content. The average Fusarium head blight indexes were 28.0% for wheat and 19.2% for triticale accessions. The percentage of Fusarium damaged kernels was also higher for wheat and came to 55.6%, while for triticale this figure was 40.2%. The average content of deoxynivalenol (DON) for wheat amounted to 11.65 mg/kg and was lower than the result for triticale which was 14.12 mg/kg. The average contents of nivalenol were similar in both cereals: 4.13 mg/kg and 5.19 mg/kg for wheat and triticale respectively. Considerable amounts of DON derivatives in the cereals were also detected. The ZEN content in the grain was 0.60 mg/kg for wheat and 0.66 mg/kg for triticale. Relationships between Fusarium head blight index, Fusarium damaged kernels and mycotoxin contents were statistically significant for wheat and mostly insignificant for triticale. Triticale proved to have less infected heads and kernels than wheat. However, the content of type B trichothecenes was higher in triticale grain than in wheat grain. PMID:27763547
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Savi, Geovana D; Cardoso, William A; Furtado, Bianca G; Bortolotto, Tiago; Zanoni, Elton T; Scussel, Rahisa; Rezende, Lucas F; Machado-de-Ávila, Ricardo A; Montedo, Oscar R K; Angioletto, Elidio
2018-03-04
Zeolites are often used as adsorbents materials and their loaded cations can be exchanged with metal ions in order to add antimicrobial properties. The aim of this study was to use the 4A zeolite and its derived ion-exchanged forms with Zn 2+ , Li + , Cu 2+ and Co 2+ in order to evaluate their antifungal properties against Fusarium graminearum, including their capacity in terms of metal ions release, conidia germination and the deoxynivalenol (DON) adsorption. The zeolites ion-exchanged with Li + , Cu 2+ , and Co 2+ showed an excellent antifungal activity against F. graminearum, using an agar diffusion method, with a zone of inhibition observed around the samples of 45.3 ± 0.6 mm, 25.7 ± 1.5 mm, and 24.7 ± 0.6 mm, respectively. Similar results using agar dilution method were found showing significant growth inhibition of F. graminearum for ion-exchanged zeolites with Zn 2+ , Li + , Cu 2+ , and Co 2+ . The fungi growth inhibition decreased as zeolite-Cu 2+ >zeolite-Li + >zeolite-Co 2+ >zeolite-Zn 2+ . In addition, the conidia germination was strongly affected by ion-exchanged zeolites. With regard to adsorption capacity, results indicate that only zeolite-Li + were capable of DON adsorption significantly (P < 0.001) with 37% at 2 mg mL -1 concentration. The antifungal effects of the ion-exchanged zeolites can be ascribed to the interactions of the metal ions released from the zeolite structure, especially for zeolite-Li + , which showed to be a promising agent against F. graminearum and its toxin.
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Van Le Thanh, Bich; Lemay, Michel; Bastien, Alexandre; Lapointe, Jérôme; Lessard, Martin; Chorfi, Younès; Guay, Frédéric
2016-05-01
Seventy-two piglets (6.0 kg BW) were randomly distributed within six different dietary treatments to evaluate the effect of deoxynivalenol (DON) and the potential of four antioxidant feed additives in mitigating the adverse effects of DON on growth performances and oxidative status. Dietary treatments were as follows: control diet 0.8 mg/kg DON; contaminated diet (DON-contaminated diet) 3.1 mg/kg DON; and four contaminated diets, each supplemented with a different antioxidant feed additive, DON + vitamins, DON + organic selenium (Se)/glutathione (GSH), DON + quercetin, and DON + COMB (vitamins + Se/GSH + quercetin from the other treatments). Although DON was the main mycotoxin in the contaminated diet, this diet also contained 1.8 mg/kg of zearalenone (ZEN). The "mycotoxin" effects therefore included the combined effect of these two mycotoxins, DON, and ZEN. The DON-ZEN ingestion did not affect growth performances, average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (G:F ratio), but partially induced oxidative stress in weaned pigs as shown by increased malondialdehyde (MDA) content in the plasma and superoxide dismutase (SOD) activity in liver (P < 0.05). However, no change in the activity of other antioxidant enzymes or GSH concentrations was observed in plasma and liver of piglets fed the DON-contaminated diet (P > 0.05). Supplementation with individual antioxidant feed additive had a limited effect in weaned pigs fed DON-ZEN-contaminated diets. Combination of antioxidants (vitamins A, C, and E, quercetin, and organic Se/GSH) reduced plasma and liver MDA content and SOD activity in liver (P < 0.05) of piglets fed DON-ZEN-contaminated diets. Furthermore, this combination also reduced MDA content in the ileum (P < 0.05), although activity of glutathione peroxidases (GPx), SOD or catalase (CAT) in the ileum was not affected by DON-ZEN contamination or antioxidant supplements. In conclusion, DON-ZEN contamination induced oxidative stress in weaned pigs and combination of antioxidant feed additives restored partially the oxidative status. Further studies will be necessary to assess whether the effects of antioxidant feed additives on oxidative status are specific when feed is contaminated with DON-ZEN.
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Gu, Wenshu; Zhu, Pei; Jiang, Donglei; He, Xingxing; Li, Yun; Ji, Jian; Zhang, Lijuan; Sun, Yange; Sun, Xiulan
2015-08-15
In this study, a novel and simple cell-based electrochemical biosensor was developed to assess the individual and combined toxicity of deoxynivalenol (DON) and zearalenone (ZEN) on BEL-7402 cells. The sensor was fabricated by modification with AuNPs, p-aminothiophenol, and folic acid in succession. The BEL-7402 cells which had a good activity were adhered on the electrode through the high affinity between the folate receptor and folic acid selectivity. We used the collagen to maintain the cell adhesion and viability. Electrochemical impedance spectroscopy (EIS) was developed to evaluate the individual and combined toxicity of DON and ZEN. Our results indicate that DON and ZEN caused a marked decrease in the cell viability in a dose-dependent manner. The value of electrochemical impedance spectroscopy decreased with the concentration of DON and ZEN in range of 0.1-20, 0.1-50 μg/ml with the detection limit as 0.03, 0.05 μg/ml, respectively, the IC50 for DON and ZEN as obtained by the proposed electrochemical method were 7.1 μg/ml and 24.6 μg/ml, respectively, and the combination of two mycotoxins appears to generate an additive response. The electrochemical cytotoxicity evaluation result was confirmed by biological assays. Compared to conventional methods, this electrochemical test is inexpensive, highly sensitive, and fast to respond, with long-term monitoring and real-time measurements. The proposed method provides a new avenue for evaluating the toxicity of mycotoxins. Copyright © 2015 Elsevier B.V. All rights reserved.
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Kovalsky, Paula; Kos, Gregor; Nährer, Karin; Schwab, Christina; Jenkins, Timothy; Schatzmayr, Gerd; Sulyok, Michael; Krska, Rudolf
2016-01-01
Global trade of agricultural commodities (e.g., animal feed) requires monitoring for fungal toxins. Also, little is known about masked and emerging toxins and metabolites. 1926 samples from 52 countries were analysed for toxins and metabolites. Of 162 compounds detected, up to 68 metabolites were found in a single sample. A subset of 1113 finished feed, maize and maize silage samples containing 57 compounds from 2012 to 2015 from 44 countries was investigated using liquid chromatography and mass spectrometry. Deoxynivalenol (DON), zearalenone (ZEN) and fumonisins showed large increases of annual medians in Europe. Within a region, distinct trends were observed, suggesting importance of local meteorology and cultivars. In 2015, median DON concentrations increased to 1400 μg·kg−1 in Austria, but were stable in Germany at 350 μg·kg−1. In 2014, enniatins occurred at median concentrations of 250 μg·kg−1 in Europe, at levels similar to DON and ZEN. The latter were frequently correlated with DON-3-glucoside and ZEN-14-sulfate. Co-occurrence of regulated toxins was frequent with e.g., enniatins, and moniliformin. Correlation was observed between DON and DON-3-glucoside and with beauvericin. Results indicate that considerably more than 25% of agricultural commodities could be contaminated with mycotoxins as suggested by FAO, although this is at least partly due to the lower limits of detection in the current survey. Observed contamination percentages ranged from 7.1 to 79% for B trichothecenes and 88% for ZEN. PMID:27929415
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A wheat ABC transporter contributes to both grain formation and mycotoxin tolerance
Walter, Stephanie; Kahla, Amal; Arunachalam, Chanemoughasoundharam; Perochon, Alexandre; Khan, Mojibur R.; Scofield, Steven R.; Doohan, Fiona M.
2015-01-01
The mycotoxin deoxynivalenol (DON) acts as a disease virulence factor for Fusarium fungi, and tolerance of DON enhances wheat resistance to Fusarium head blight (FHB) disease. Two variants of an ATP-binding cassette (ABC) family C transporter gene were cloned from DON-treated wheat mRNA, namely TaABCC3.1 and TaABCC3.2. These represent two of three putative genes identified on chromosomes 3A, 3B, and 3D of the wheat genome sequence. Variant TaABCC3.1 represents the DON-responsive transcript previously associated with DON resistance in wheat. PCR-based mapping and in silico sequence analyses located TaABCC3.1 to the short arm of wheat chromosome 3B (not within the FHB resistance quantitative trait locus Fhb1). In silico analyses of microarray data indicated that TaABCC3 genes are expressed in reproductive tissue and roots, and in response to the DON producer Fusarium graminearum. Gene expression studies showed that TaABCC3.1 is activated as part of the early host response to DON and in response to the FHB defence hormone jasmonic acid. Virus-induced gene silencing (VIGS) confirmed that TaABCC3 genes contributed to DON tolerance. VIGS was performed using two independent viral construct applications: one specifically targeted TaABCC3.1 for silencing, while the other targeted this gene and the chromosome 3A homeologue. In both instances, VIGS resulted in more toxin-induced discoloration of spikelets, compared with the DON effects in non-silenced spikelets at 14 d after toxin treatment (≥2.2-fold increase, P<0.05). Silencing by both VIGS constructs enhanced head ripening, and especially so in DON-treated heads. VIGS of TaABCC3 genes also reduced the grain number by more than 28% (P<0.05), both with and without DON treatment, and the effects were greater for the construct that targeted the two homeologues. Hence, DON-responsive TaABCC3 genes warrant further study to determine their potential as disease resistance breeding targets and their function in grain formation and ripening. PMID:25732534
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Wetterhorn, Karl M; Gabardi, Kaitlyn; Michlmayr, Herbert; Malachova, Alexandra; Busman, Mark; McCormick, Susan P; Berthiller, Franz; Adam, Gerhard; Rayment, Ivan
2017-12-19
Family 1 UDP-glycosyltransferases (UGTs) in plants primarily form glucose conjugates of small molecules and, besides other functions, play a role in detoxification of xenobiotics. Indeed, overexpression of a barley UGT in wheat has been shown to control Fusarium head blight, which is a plant disease of global significance that leads to reduced crop yields and contamination with trichothecene mycotoxins such as deoxynivalenol (DON), T-2 toxin, and many other structural variants. The UGT Os79 from rice has emerged as a promising candidate for inactivation of mycotoxins because of its ability to glycosylate DON, nivalenol, and hydrolyzed T-2 toxin (HT-2). However, Os79 is unable to modify T-2 toxin (T-2), produced by pathogens such as Fusarium sporotrichioides and Fusarium langsethii. Activity toward T-2 is desirable because it would allow a single UGT to inactivate co-occurring mycotoxins. Here, the structure of Os79 in complex with the products UDP and deoxynivalenol 3-O-glucoside is reported together with a kinetic analysis of a broad range of trichothecene mycotoxins. Residues associated with the trichothecene binding pocket were examined by site-directed mutagenesis that revealed that trichothecenes substituted at the C4 position, which are not glycosylated by wild-type Os79, can be accommodated in the binding pocket by increasing its volume. The H122A/L123A/Q202L triple mutation, which increases the volume of the active site and attenuates polar contacts, led to strong and equivalent activity toward trichothecenes with C4 acetyl groups. This mutant enzyme provides the broad specificity required to control multiple toxins produced by different Fusarium species and chemotypes.
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Mossoba, M M; Adams, S; Roach, J A; Trucksess, M W
1996-01-01
Gas chromatography/matrix isolation/Fourier transform infrared (GC/MI/FTIR) spectroscopy and GC/mass spectrometry (MS) were used to confirm the identities of trimethylsilyl (TMS) derivatives of trichothecene mycotoxins in naturally contaminated grains. Infrared spectral bands observed in the fingerprint region were unique for 10 trichothecene standards. Characteristic absorption bands were observed for the ester (near 1750 cm-1) and ketone (near 1700 cm-1) carbonyl stretching vibrations, the acetate CH3 symmetric bend (1370 cm-1), the epoxide ring (1262 cm-1), the trimethylsilyl CH3 in-plane deformation (1253 cm-1), the ester (O)C-O asymmetric stretching vibration (near 1244 cm-1), and several other bands including intense features due to the TMS function. Infrared bands observed under cryogenic matrix isolation conditions were compared with those found at room temperature in a potassium bromide matrix for 5 of these standards. Identities of deoxynivalenol (DON) from barley and mixed feed, nivalenol from wheat and barley, and DON and fusarenon-x from sweet corn were confirmed by comparison of their infrared spectral bands with those of standards. The identity of DON in the same test samples of sweet corn was confirmed further by GC/MS. GC/MS was also used to quantitate the levels of DON (67-455 ppm) in sweet corn test samples.
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Tola, Siriporn; Bureau, Dominique P.; Hooft, Jamie M.; Beamish, Frederick W. H.; Sulyok, Michael; Krska, Rudolf; Encarnação, Pedro; Petkam, Rakpong
2015-01-01
An 8-week feeding trial was conducted to examine effects of wheat naturally contaminated with Fusarium mycotoxins (deoxynivalenol, DON 41 mg·kg−1) on growth performance and selected health indices of red tilapia (Oreochromis niloticus × O. mossambicus; initial weight = 4.3 g/fish). Five experimental diets were formulated by replacement of clean wheat with naturally contaminated wheat resulting in graded levels of DON and zearalenone (ZEN) (Diet 1 0.07/0.01, Diet 2 0.31/0.09, Diet 3 0.50/0.21, Diet 4 0.92/0.37 and Diet 5 1.15/0.98 mg·kg−1). Groups of 50 fish were randomly allocated into each of 20 aquaria and fed to near-satiety for eight weeks. Growth rate, feed intake and feed efficiency of fish fed the experimental diets decreased linearly with increasing levels of Fusarium mycotoxins (p < 0.05). Although growth depression was associated with feeding diets naturally contaminated with Fusarium mycotoxins, especially DON, no biochemical and histopathological parameters measured in blood and liver appeared affected by Fusarium mycotoxin concentrations of diets (p > 0.05). Though there was no clear evidence of overt DON toxicity to red tilapia, it is recommended that feed ingredients should be screened for Fusarium mycotoxin contamination to ensure optimal growth performance. PMID:26035489
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Purification, immunotoxic effects, and cellular uptake of trichothecene mycotoxins
DOE Office of Scientific and Technical Information (OSTI.GOV)
Witt, M.F.
1989-01-01
Studies were carried out to better understand how the trichothecenes alter immune function in animals and humans. Deoxynivalenol (DON) was purified for use in animal feeding studies. Dietary exposure to DON for 8 weeks altered the serum immunoglobulin profile in mice and decreased the splenic plaque-forming cell response to the antigen sheep red blood cells. The uptake of ({sup 3}H)T-2 toxin by a murine B-cell hybridoma was studied in order to learn more about the way in which trichothecenes interact with immune cells. A simple procedure was developed for the laboratory production and purification of gram quantities of crystalline DON.more » When Fusarium graminearum R6576 was grown on rice, concentrations of 600 to 700 ppm DON accumulated after 13 to 18 days of incubation. A DON derivative, 15-acetylDON, was also found at concentrations of 100 to 300 ppm after 7 to 10 days. DON was purified from crude culture extracts by water-saturated silica gel chromatography. Alpha-({sup 3}H)T-2 toxin of 99% chemical and radiochemical purity was prepared for use in uptake studies. Both the rate of uptake of ({sup 3}H)T-2 toxin by hybridomas and the time required for accumulation of ({sup 3}H)T-2 to reach equilibrium were proportional to the concentration of ({sup 3}H)T-2. ({sup 3}H)T-2 toxin accumulated by hybridomas was proportional to the concentration of ({sup 3}H)T-2 between 10{sup {minus}8} and 10{sup {minus}3} M. The rate of uptake of ({sup 3}H)jT-2 toxin by hybridomas was inhibited by the trichothecenes T-2 toxin, DON, verrucarin A, and roridin A, as well as the antibiotic anisomycin. The kinetics and concentration dependence of accumulation, along with the inhibition patterns, suggest that uptake of ({sup 3}H)T-2 toxin by hybridomas is mediated by binding of toxin to ribosomes.« less
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NASA Astrophysics Data System (ADS)
Kong, Dezhao; Liu, Liqiang; Song, Shanshan; Suryoprabowo, Steven; Li, Aike; Kuang, Hua; Wang, Libing; Xu, Chuanlai
2016-02-01
A semi-quantitative and quantitative multi-immunochromatographic (ICA) strip detection assay was developed for the simultaneous detection of twenty types of mycotoxins from five classes, including zearalenones (ZEAs), deoxynivalenols (DONs), T-2 toxins (T-2s), aflatoxins (AFs), and fumonisins (FBs), in cereal food samples. Sensitive and specific monoclonal antibodies were selected for this assay. The semi-quantitative results were obtained within 20 min by the naked eye, with visual limits of detection for ZEAs, DONs, T-2s, AFs and FBs of 0.1-0.5, 2.5-250, 0.5-1, 0.25-1 and 2.5-10 μg kg-1, and cut-off values of 0.25-1, 5-500, 1-10, 0.5-2.5 and 5-25 μg kg-1, respectively. The quantitative results were obtained using a hand-held strip scan reader, with the calculated limits of detection for ZEAs, DONs, T-2s, AFs and FBs of 0.04-0.17, 0.06-49, 0.15-0.22, 0.056-0.49 and 0.53-1.05 μg kg-1, respectively. The analytical results of spiked samples were in accordance with the accurate content in the simultaneous detection analysis. This newly developed ICA strip assay is suitable for the on-site detection and rapid initial screening of mycotoxins in cereal samples, facilitating both semi-quantitative and quantitative determination.A semi-quantitative and quantitative multi-immunochromatographic (ICA) strip detection assay was developed for the simultaneous detection of twenty types of mycotoxins from five classes, including zearalenones (ZEAs), deoxynivalenols (DONs), T-2 toxins (T-2s), aflatoxins (AFs), and fumonisins (FBs), in cereal food samples. Sensitive and specific monoclonal antibodies were selected for this assay. The semi-quantitative results were obtained within 20 min by the naked eye, with visual limits of detection for ZEAs, DONs, T-2s, AFs and FBs of 0.1-0.5, 2.5-250, 0.5-1, 0.25-1 and 2.5-10 μg kg-1, and cut-off values of 0.25-1, 5-500, 1-10, 0.5-2.5 and 5-25 μg kg-1, respectively. The quantitative results were obtained using a hand-held strip scan reader, with the calculated limits of detection for ZEAs, DONs, T-2s, AFs and FBs of 0.04-0.17, 0.06-49, 0.15-0.22, 0.056-0.49 and 0.53-1.05 μg kg-1, respectively. The analytical results of spiked samples were in accordance with the accurate content in the simultaneous detection analysis. This newly developed ICA strip assay is suitable for the on-site detection and rapid initial screening of mycotoxins in cereal samples, facilitating both semi-quantitative and quantitative determination. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr09171c
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Xia, Shuang; Zhu, Pei; Pi, Fuwei; Zhang, Yinzhi; Li, Yun; Wang, Jiasheng; Sun, Xiulan
2017-11-15
A simple and convenient cell-based electrochemical biosensor was developed to assess the individual and combined toxicity of deoxynivalenol (DON), zearalenone (ZEN), and Aflatoxin B 1 (AFB 1 ) on Hep G2 cells. The sensor was modified in succession with AuNPs (gold nanoparticles), cysteamine, and laminin. The cells interacting with laminin formed tight cell-to-electrode contacts, and collagen was used to maintain cell adhesion and viability. Electrochemical impedance spectroscopy (EIS) was developed to evaluate mycotoxin toxicity. Experimental results show that DON, ZEN, and AFB 1 caused a significant decrease in cell viability in a dose dependent manner. The EIS value decreased with concentrations of DON, ZEN, and AFB 1 in the range of 0.01-20, 0.1-50, and 0.1-3.5μg/mL, and IC 50 obtained using the developed method was 48.5, 59.0, and 3.10μg/mL, respectively. A synergistic effect was observed between DON and ZEN, an additive effect was observed between DON and AFB 1 , and an antagonism effect was found in the binary mixtures of ZEN and AFB 1 and ternary mixtures. These results were confirmed via CCK-8 assay. Utilizing SEM, we found that cells treated with mycotoxins caused significant changes in cell morphology, thus lessening cell adsorption and impedance reduction. Biological assay indicated that EIS patterns correlated with [Ca 2+ ] i concentrations and apoptosis and necrotic cells ratios, thus effecting electrochemical signals. This method is simpler, more convenient, sensitive, and has a quicker response rate than most conventional cytotoxicity evaluation methods. Copyright © 2017. Published by Elsevier B.V.
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Wang, Jian-Hua; Li, He-Ping; Qu, Bo; Zhang, Jing-Bo; Huang, Tao; Chen, Fang-Fang; Liao, Yu-Cai
2008-01-01
Fusarium graminearum clade pathogens cause Fusarium head blight (FHB) or scab of wheat and other small cereal grains, producing different kinds of trichothecene mycotoxins that are detrimental to human and domestic animals. Type B trichothecene mycotoxins such as deoxynivalenol, 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON) and nivalenol (NIV) are the principal Fusarium mycotoxins reported in China, as well as in other countries. A genomic polymerase chain reaction (PCR) to predict chemotypes was developed based on the structural gene sequences of Tri13 genes involved in trichothecene mycotoxin biosynthesis pathways. A single pair of primers derived from the Tri13 genes detected a 583 bp fragment from 15-AcDON-chemotypes, a 644 bp fragment from 3-AcDON-chemotypes and an 859 bp fragment from NIV-producing strains. Fusarium strains from China, Nepal, USA and Europe were identified by this method, revealing their mycotoxin chemotypes identical to that obtained by chemical analyses of HPLC or GC/MS and other PCR assays. The mycotoxin chemotype-specific fragments were amplified from a highly variable region located in Tri13 genes with three deletions for 15-AcDON-chemotypes, two deletions for 3-AcDON-chemotypes and no deletion for NIV-producers. This PCR assay generated a single amplicon and thus should be more reliable than other PCR-based assays that showed the absence or presence of a PCR fragment since these assays may generate false-negative results. The results with strains from several different countries as well as from different hosts further indicated that this method should be globally applicable. This is a rapid, reliable and cost-effective method for the identification of type B trichothecene mycotoxin chemotypes in Fusarium species and food safety controls. PMID:19330088
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Masci, Stefania; Laino, Paolo; Janni, Michela; Botticella, Ermelinda; Di Carli, Mariasole; Benvenuto, Eugenio; Danieli, Pier Paolo; Lilley, Kathryn S; Lafiandra, Domenico; D'Ovidio, Renato
2015-04-22
Fusarium head blight, caused by the fungus Fusarium graminearum, has a detrimental effect on both productivity and qualitative properties of wheat. To evaluate its impact on wheat flour, we compared its effect on quality-related parameters between a transgenic bread wheat line expressing a bean polygalacturonase inhibiting protein (PGIP) and its control line. We have compared metabolic proteins, the amounts of gluten proteins and their relative ratios, starch content, yield, extent of pathogen contamination, and deoxynivalenol (DON) accumulation. These comparisons showed that Fusarium significantly decreases the amount of starch in infected control plants, but not in infected PGIP plants. The flour of PGIP plants contained also a lower amount of pathogen biomass and DON accumulation. Conversely, both gluten and metabolic proteins were not significantly influenced either by the transgene or by fungal infection. These results indicate that the transgenic PGIP expression reduces the level of infection, without changing significantly the wheat seed proteome and other quality-related parameters.
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Regulation of metabolic products and gene expression in Fusarium asiaticum by agmatine addition.
Suzuki, Tadahiro; Kim, Young-Kyung; Yoshioka, Hifumi; Iwahashi, Yumiko
2013-05-01
The metabolic products resulting from the cultivation of F. asiaticum in agmatine were identified using capillary electrophoresis-time of flight mass spectrometry. Glyoxylic acid was detected from fungal cultures grown in agmatine, while it was absent in control cells. The abundance of other metabolic products of the glycolytic pathway also increased because of agmatine; however, there was no increase in the amounts of pyruvic acid or metabolites from the tricarboxylic acid cycle. Moreover, gene expression levels within Fusarium asiaticum exposed to agmatine were analyzed by DNA microarray. Changes in gene expression levels directed the changes in metabolic products. Our results suggest that acetyl coenzyme A, which is a starting substrate for the biosynthesis of deoxynivalenol (DON), was simultaneously produced by activated β-oxidation. Furthermore, the content of 4-aminobutyrate (GABA) was increased in the agmatine addition culture medium. GABA can be synthesized from agmatine through putrescine and might influence the regulation of DON-related genes.
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Mycotoxins in commercial dry pet food in China.
Shao, Manyu; Li, Li; Gu, Zuli; Yao, Ming; Xu, Danning; Fan, Wentao; Yan, Liping; Song, Suquan
2018-05-10
The aim of this study was to investigate the occurrence of the most common mycotoxins in commercial dry dog food using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Aflatoxin B1 (AFB1), aflatoxin G1 (AFG1), zearalenone (ZEN), deoxynivalenol (DON), T-2, beauvericin (BEA), citrinin (CIT), ochratoxin A (OTA), fumonisin B1 (FB1) were included in this study. The results showed that all these analytes could be found in the samples. Furthermore, only one sample was found free of mycotoxins contamination. All other samples (96.9%) were contaminated by at least three different types of mycotoxins. Among these mycotoxins, DON, ZEN, AFB1, FB1, CIT and BEA exhibited relatively high incidence, with occurrence rates of 78.1%, 62.5%, 87.5%, 93.8%, 68.8 and 96.9%. Furthermore, it is worth noting that AFB1 concentration in all AFB1 positive samples exceeded the maximum limits set by the EU, with concentrations ranging from 30.3 µg/kg to 242.7 µg/kg.
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Musa, Tomke; Bänziger, Irene; Kägi, Andreas; Bucheli, Thomas D.; Wettstein, Felix E.; Forrer, Hans-Rudolf
2017-01-01
To assess the occurrence of Fusarium toxins in wheat in Switzerland, an eight-year survey was conducted by analysing a total of 686 harvest samples from growers using LC-MS/MS. Between 2007 and 2010, 527 samples were obtained from 17 cantons. Between 2011 and 2014, 159 samples were collected from the canton Berne. The most frequent toxins detected were deoxynivalenol (DON), zearalenone (ZEA) and nivalenol (NIV). The overall mean DON content in all samples was 607 µg/kg, and 11% exceeded the European limit for unprocessed cereals for foodstuffs (1250 µg/kg). For ZEA (mean 39 µg/kg), 7% exceeded the respective limit (100 µg/kg), and the mean content of NIV (no limit established) was 15 µg/kg. Between the years, the ratio of mycotoxin-contaminated samples ranged between 52% and 98% for DON, 9% and 43% for ZEA and 0% and 49% for NIV. The yearly mean contents varied substantially between 68 and 1310 µg/kg for DON, 5 and 56 µg/kg for ZEA and 6 and 29 µg/kg for NIV. The geographic origin showed a significant effect on DON and ZEA contamination, but was inconsistent between the years. This study has shown that the majority of Swiss-produced wheat is, in terms of Fusarium toxins, fit for human consumption and feed purposes. Nevertheless, depending on the year, high toxin contents can be expected, an issue that growers, cereal collection centres and the food industry have to deal with to ensure food and feed safety. PMID:28792467
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Vogelgsang, Susanne; Musa, Tomke; Bänziger, Irene; Kägi, Andreas; Bucheli, Thomas D; Wettstein, Felix E; Pasquali, Matias; Forrer, Hans-Rudolf
2017-08-09
To assess the occurrence of Fusarium toxins in wheat in Switzerland, an eight-year survey was conducted by analysing a total of 686 harvest samples from growers using LC-MS/MS. Between 2007 and 2010, 527 samples were obtained from 17 cantons. Between 2011 and 2014, 159 samples were collected from the canton Berne. The most frequent toxins detected were deoxynivalenol (DON), zearalenone (ZEA) and nivalenol (NIV). The overall mean DON content in all samples was 607 µg/kg, and 11% exceeded the European limit for unprocessed cereals for foodstuffs (1250 µg/kg). For ZEA (mean 39 µg/kg), 7% exceeded the respective limit (100 µg/kg), and the mean content of NIV (no limit established) was 15 µg/kg. Between the years, the ratio of mycotoxin-contaminated samples ranged between 52% and 98% for DON, 9% and 43% for ZEA and 0% and 49% for NIV. The yearly mean contents varied substantially between 68 and 1310 µg/kg for DON, 5 and 56 µg/kg for ZEA and 6 and 29 µg/kg for NIV. The geographic origin showed a significant effect on DON and ZEA contamination, but was inconsistent between the years. This study has shown that the majority of Swiss-produced wheat is, in terms of Fusarium toxins, fit for human consumption and feed purposes. Nevertheless, depending on the year, high toxin contents can be expected, an issue that growers, cereal collection centres and the food industry have to deal with to ensure food and feed safety.
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Impact of competitive fungi on Trichothecene production by Fusarium graminearum.
Cooney, J M; Lauren, D R; di Menna, M E
2001-01-01
Bioassays were used to determine the production of the trichothecene mycotoxin, deoxynivalenol (DON), by two isolates of Fusarium graminearum when grown in association with potentially competitive fungi and an antifungal chemical, 6-pentyl-alpha-pyrone (6PAP). The presence of 6PAP in the culture medium reduced DON production by as much as 80%, but this effect was reduced for the F. graminearum isolate that most efficiently metabolized the added 6PAP. A 6PAP-producing Trichoderma isolate grown in a competition assay system with the F. graminearum isolates was also able to substantially reduce DON production. When Fusarium isolates (F. crookwellense, F. culmorum, F. subglutinans, F. poae, F. equiseti, F. avenaceum, and F. sambucinum), which co-occur with F. graminearum in New Zealand maize plants (Zea mays), were grown in competition assays, the effect on DON production was variable. However, all isolates of F. subglutinans tested were shown to cause reductions in DON production (by 13-76%, mean = 62%). F. subglutinans frequently co-occurs with F. graminearum, but its presence can vary with location and time of the season. When the competitive fungus tested was also a trichothecene producer (e.g., of nivalenol), both toxins were produced in the assay medium. The results indicate that mycotoxin production by F. graminearum can be affected by the presence of particular competitive fungi. These results have implications for an ecological understanding of pathogenicity and of mycotoxin accumulation in plants. Early establishment of F. subglutinans, for example, may act as a biological control mechanism providing a temporary protection against invasion by more commonly toxigenic fusaria such as F. graminearum.
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Current Situation of Mycotoxin Contamination and Co-occurrence in Animal Feed—Focus on Europe
Streit, Elisabeth; Schatzmayr, Gerd; Tassis, Panagiotis; Tzika, Eleni; Marin, Daniela; Taranu, Ionelia; Tabuc, Cristina; Nicolau, Anca; Aprodu, Iuliana; Puel, Olivier; Oswald, Isabelle P.
2012-01-01
Mycotoxins are secondary metabolites produced by fungi especially those belonging to the genus Aspergillus, Penicillum and Fusarium. Mycotoxin contamination can occur in all agricultural commodities in the field and/or during storage, if conditions are favourable to fungal growth. Regarding animal feed, five mycotoxins (aflatoxins, deoxynivalenol, zearalenone, fumonisins and ochratoxin A) are covered by EU legislation (regulation or recommendation). Transgressions of these limits are rarely observed in official monitoring programs. However, low level contamination by Fusarium toxins is very common (e.g., deoxynivalenol (DON) is typically found in more than 50% of the samples) and co-contamination is frequently observed. Multi-mycotoxin studies reported 75%–100% of the samples to contain more than one mycotoxin which could impact animal health at already low doses. Co-occurrence of mycotoxins is likely to arise for at least three different reasons (i) most fungi are able to simultaneously produce a number of mycotoxins, (ii) commodities can be contaminated by several fungi, and (iii) completed feed is made from various commodities. In the present paper, we reviewed the data published since 2004 concerning the contamination of animal feed with single or combinations of mycotoxins and highlighted the occurrence of these co-contaminations. PMID:23162698
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2010-01-01
Background The fungal pathogen Fusarium graminearum causes Fusarium Head Blight (FHB) disease on wheat which can lead to trichothecene mycotoxin (e.g. deoxynivalenol, DON) contamination of grain, harmful to mammalian health. DON is produced at low levels under standard culture conditions when compared to plant infection but specific polyamines (e.g. putrescine and agmatine) and amino acids (e.g. arginine and ornithine) are potent inducers of DON by F. graminearum in axenic culture. Currently, host factors that promote mycotoxin synthesis during FHB are unknown, but plant derived polyamines could contribute to DON induction in infected heads. However, the temporal and spatial accumulation of polyamines and amino acids in relation to that of DON has not been studied. Results Following inoculation of susceptible wheat heads by F. graminearum, DON accumulation was detected at two days after inoculation. The accumulation of putrescine was detected as early as one day following inoculation while arginine and cadaverine were also produced at three and four days post-inoculation. Transcripts of ornithine decarboxylase (ODC) and arginine decarboxylase (ADC), two key biosynthetic enzymes for putrescine biosynthesis, were also strongly induced in heads at two days after inoculation. These results indicated that elicitation of the polyamine biosynthetic pathway is an early response to FHB. Transcripts for genes encoding enzymes acting upstream in the polyamine biosynthetic pathway as well as those of ODC and ADC, and putrescine levels were also induced in the rachis, a flower organ supporting DON production and an important route for pathogen colonisation during FHB. A survey of 24 wheat genotypes with varying responses to FHB showed putrescine induction is a general response to inoculation and no correlation was observed between the accumulation of putrescine and infection or DON accumulation. Conclusions The activation of the polyamine biosynthetic pathway and putrescine in infected heads prior to detectable DON accumulation is consistent with a model where the pathogen exploits the generic host stress response of polyamine synthesis as a cue for production of trichothecene mycotoxins during FHB disease. However, it is likely that this mechanism is complicated by other factors contributing to resistance and susceptibility in diverse wheat genetic backgrounds. PMID:21192794
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Incidence of Fusarium Species and Mycotoxins in Silage Maize
Eckard, Sonja; Wettstein, Felix E.; Forrer, Hans-Rudolf; Vogelgsang, Susanne
2011-01-01
Maize is frequently infected by the Fusarium species producing mycotoxins. Numerous investigations have focused on grain maize, but little is known about the Fusarium species in the entire plant used for silage. Furthermore, mycotoxins persist during the ensiling process and thus endanger feed safety. In the current study, we analyzed 20 Swiss silage maize samples from growers’ fields for the incidence of Fusarium species and mycotoxins. The species spectrum was analyzed morphologically and mycotoxins were measured by LC-MS/MS. A pre-harvest visual disease rating showed few disease symptoms. In contrast, the infection rate of two-thirds of the harvest samples ranged from 25 to 75% and twelve different Fusarium species were isolated. The prevailing species were F. sporotrichioides, F. verticillioides and F. graminearum. No infection specificity for certain plant parts was observed. The trichothecene deoxynivalenol (DON) was found in each sample (ranging from 780 to 2990 µg kg−1). Other toxins detected in descending order were zearalenone, further trichothecenes (nivalenol, HT-2 and T-2 toxin, acetylated DON) and fumonisins. A generalized linear regression model containing the three cropping factors harvest date, pre-precrop and seed treatment was established, to explain DON contamination of silage maize. Based on these findings, we suggest a European-wide survey on silage maize. PMID:22069750
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Devreese, Mathias; Girgis, George N; Tran, Si-Trung; De Baere, Siegrid; De Backer, Patrick; Croubels, Siska; Smith, Trevor K
2014-01-01
An experiment was conducted to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins and a yeast derived glucomannan mycotoxin adsorbent (GMA) on selected specific and non-specific parameters in turkey poults. Two hundred and forty 1-day-old male turkey poults were fed the experimental diets for twelve weeks. Experimental diets were formulated with control grains, control grains+0.2% GMA, naturally-contaminated grains, or naturally-contaminated grains+0.2% GMA. Deoxynivalenol (DON) was the major contaminant of the contaminated grains and concentrations varied from 4.0 to 6.5 mg/kg in the contaminated diets. Non-specific parameters measured included: performance parameters, plasma biochemistry profiles, morphometry and CD8(+) T-lymphocyte counts in the duodenum. Plasma concentrations of DON and de-epoxydeoxynivalenol (DOM-1) were used as specific parameters. Performance parameters and plasma biochemistry were altered by the feeding of contaminated diets and GMA but this was not consistent throughout the trial. The feeding of contaminated diets reduced duodenal villus height and apparent villus surface area. This effect was prevented by GMA supplementation. The feeding of contaminated diets elevated total duodenal CD8(+) T-lymphocyte counts but this effect was not prevented by GMA. No significant differences were seen in plasma concentrations of DON and DOM-1 comparing birds fed contaminated and contaminated+GMA diets suggesting that GMA did not prevent DON absorption under these conditions. Copyright © 2013 Elsevier Ltd. All rights reserved.
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Fusarium mycotoxin content of UK organic and conventional oats.
Edwards, S G
2009-07-01
Every year between 2002 and 2005 approximately 100 samples of oats from fields of known agronomy were analysed by GC/MS for 10 trichothecenes: deoxynivalenol (DON), nivalenol, 3-acetylDON, 15-acetylDON, fusarenone X, T-2 toxin (T2), HT-2 toxin (HT2), diacetoxyscirpenol, neosolaniol and T-2 triol. Samples were also analysed for moniliformin and zearalenone by HPLC. Of the 10 trichothecenes analysed from 458 harvest samples of oat only three, 15-acetylDON, fusarenone X and diacetoxyscirpenol, were not detected. Moniliformin and zearalenone were absent or rarely detected, respectively. HT2 and T2 were the most frequently detected fusarium mycotoxins, present above the limit of quantification (10 microg kg(-1)) in 92 and 84% of samples, respectively, and were usually present at the highest concentrations. The combined mean and median for HT2 and T2 (HT2 + T2) was 570 and 213 microg kg(-1), respectively. There were good correlations between concentrations of HT2 and all other type A trichothecenes detected (T2, T2 triol and neosolaniol). Year and region had a significant effect on HT2 + T2 concentration. There was also a highly significant difference between HT2 + T2 content in organic and conventional samples, with the predicted mean for organic samples five times lower than that of conventional samples. This is the largest difference reported for any mycotoxin level in organic and conventional cereals. No samples exceeded the legal limits for DON or zearalenone in oats intended for human consumption. Legislative limits for HT2 and T2 are currently under consideration by the European Commission. Depending on the limits set for unprocessed oats intended for human consumption, the levels detected here could have serious consequences for the UK oat-processing industry.
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Regulation of IL-8 promoter activity by verrucarin A in human monocytic THP-1 cells.
Liu, Jun; Simmons, Steve O; Pei, Ruoting
2014-01-01
Macrocyclic trichothecenes have been frequently detected in fungi in water-damaged buildings and exhibited higher toxicity than the well-studied trichothecenes; however, the mechanism underlying their toxicity has been poorly understood. In this study, transcriptional regulation of the cytokine interleukin (IL)-8 by a macrocyclic trichothecene, verrucarin A (VA), in human monocytic THP-1 cells is reported. Consistent with previous findings, VA was 100-fold more cytotoxic than deoxynivalenol (DON), while ochratoxin A (OA) was not cytotoxic. In cells transduced with the wild-type IL-8 promoter luciferase construct, VA induced a biphasic dose response composed of an upregulation of luciferase expression at low concentrations of 0.01-1 ng/ml and a downregulation at high levels of 10 ng/ml and higher. In contrast, DON induced a sigmoid-shaped dose response with the EC50 of 11.6 ng/ml, while OA did not markedly affect the IL-8 expression. When cells were transduced with IL-8 promoter with a mutation of transcription factor nuclear factor-κB (NF-κB)-binding site, VA (1 ng/ml), DON (1000 ng/ml), and tumor necrosis factor (TNF) α (20 ng/ml)-induced luciferase expression were impaired. In addition, the NF-κB inhibitor caffeic acid phenethyl ester inhibited VA-, DON-, and TNFα-induced luciferase expression. Mutation of the CCAAT/enhancer-binding protein (CEBP) β binding site of the IL-8 promoter affected only DON-, but not VA- and TNFα-induced luciferase expression. Taken together, these results suggested that VA activated IL-8 promoter via an NF-κB-dependent, but not CEBPβ-dependent, pathway in human monocytes.
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Warth, Benedikt; Petchkongkaew, Awanwee; Sulyok, Michael; Krska, Rudolf
2014-01-01
Human exposures to mycotoxins through dietary intake are a major health hazard and may result in various pathophysiological effects. Although Thailand is a country at increased risk due to its climatic conditions, no comprehensive dataset is available to perform proper exposure assessment of its population with regard to mycotoxins. Therefore, this pilot study was conducted to investigate and evaluate the exposure levels of major mycotoxins (aflatoxin B₁, ochratoxin A, fumonisins, zearalenone and trichothecenes). Sixty first-morning urine samples were collected from healthy volunteers who live in the Bangkok metropolitan area and surrounding provinces (Pathumthani, Nonthaburi, Samutprakarn and Samutsakorn). Urine samples were analysed by a LC-MS/MS-based multi-biomarker method following a so-called 'dilute and shoot' approach. Results generally indicated low mycotoxin exposures in most individuals through the determination of the four biomarkers that were detected in urine samples, i.e. aflatoxin M₁, ochratoxin A (OTA), as well as the deoxynivalenol (DON) metabolites DON-3-glucuronide and DON-15-glucuronide in 10 of 60 individuals. The maximum concentrations were used to estimate the daily intake confirming that none of the individuals exceeded the tolerable daily intake (TDI) of DON (maximum 26% of TDI) or OTA (maximum 22% of TDI). However, the maximum exposure of aflatoxin B₁, estimated to be 0.91 µg (kg bw)⁻¹ day⁻¹, should raise some concerns and suggests further studies utilising a more sensitive method. Low exposure to Fusarium toxins was also confirmed by the absence of zearalenone, α-zearalanol, β-zearalanol and zearalenone-14-glucuronide as well as T-2 toxin, HT-2 toxin, nivalenol and free DON. This is the first multi-mycotoxin biomarker study performed in Southeast Asia.
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He, Yi; Ahmad, Dawood; Zhang, Xu; Zhang, Yu; Wu, Lei; Jiang, Peng; Ma, Hongxiang
2018-04-19
Fusarium head blight (FHB), a devastating disease in wheat worldwide, results in yield loses and mycotoxin, such as deoxynivalenol (DON), accumulation in infected grains. DON also facilitates the pathogen colonization and spread of FHB symptoms during disease development. UDP-glycosyltransferase enzymes (UGTs) are known to contribute to detoxification and enhance FHB resistance by glycosylating DON into DON-3-glucoside (D3G) in wheat. However, a comprehensive investigation of wheat (Triticum aestivum) UGT genes is still lacking. In this study, we carried out a genome-wide analysis of family-1 UDP glycosyltransferases in wheat based on the PSPG conserved box that resulted in the identification of 179 putative UGT genes. The identified genes were clustered into 16 major phylogenetic groups with a lack of phylogenetic group K. The UGT genes were invariably distributed among all the chromosomes of the 3 genomes. At least 10 intron insertion events were found in the UGT sequences, where intron 4 was observed as the most conserved intron. The expression analysis of the wheat UGT genes using both online microarray data and quantitative real-time PCR verification suggested the distinct role of UGT genes in different tissues and developmental stages. The expression of many UGT genes was up-regulated after Fusarium graminearum inoculation, and six of the genes were further verified by RT-qPCR. We identified 179 UGT genes from wheat using the available sequenced wheat genome. This study provides useful insight into the phylogenetic structure, distribution, and expression patterns of family-1 UDP glycosyltransferases in wheat. The results also offer a foundation for future work aimed at elucidating the molecular mechanisms underlying the resistance to FHB and DON accumulation.
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Goyarts, Tanja; Brüssow, Klaus-Peter; Valenta, Hana; Tiemann, Ute; Jäger, Kathrin; Dänicke, Sven
2010-05-01
Six pregnant sows of 180.6 ± 5.6 kg were fed either a Fusarium-contaminated (4.42 mg DON and 48.3 µg ZON per kg, DON per os, n = 3) or a control diet (0.15 mg DON and 5 µg ZON/kg) in the period of days 63 and 70 of gestation. On day 63 of gestation, sows fed the control diet were implanted with an intraperitoneal osmotic minipump (delivery rate of 10 µL/h, for 7 days) containing 50 mg pure (98%) DON in 2 ml 50% DMSO (DON ip, n = 3). Frequent plasma samples were taken to estimate the kinetics after oral and ip DON exposure. The intended continuous delivery of DON by the intraperitoneal minipump could not be shown, as there was a plasma peak (Cmax) of 4.2-6.4 ng DON/mL either immediately (sow IP-2+3) or 2.5 h (sow IP-1) after implantation of the pump followed by a one-exponential decline with a mean half-time (t1/2) of 1.75-4.0 h and only negligible DON plasma concentrations after 12 h. Therefore, the DON ip exposure has to be regarded as one single dose 1 week before termination of experiment. The DON per os sows showed a mean basis level (after achieving a steady state) of DON plasma concentration of about 6-8 ng/mL, as also indicated by the plasma DON concentration at the termination of the experiment. On day 70, caesarean section was carried out, the fetuses were killed immediately after birth, and samples of plasma, urine, and bile were taken to analyze the concentration of DON and its metabolite de-epoxy-DON. At necropsy there were no macroscopic lesions observed in any organ of either sows or piglets. Histopathological evaluation of sows liver and spleen revealed no alterations. The proliferation rate of peripheral blood mononuclear cells (PBMC) with or without stimulation was not affected by the kind of DON treatment. The exposure of pregnant sows at mid-gestation (days 63-70, period of organogenesis) to a Fusarium toxin-contaminated diet (4.42 mg DON and 0.048 mg ZON per kg) or pure DON via intraperitoneal osmotic minipump did not cause adverse effects on health, fertility, maintenance of pregnancy, and performance of sows and their fetuses. However, DON was detected in fetus plasma, indicating that this toxin can pass the placental barrier and may cause changes in the proportion of white blood cells (lower monocyte and neutrophil and higher lymphocyte proportion in DON per os fetuses).
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Broekaert, N; Devreese, M; De Mil, T; Fraeyman, S; De Baere, S; De Saeger, S; De Backer, P; Croubels, S
2014-11-15
This study aims to develop an LC-MS/MS method allowing the determination of 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, deoxynivalenol and its main in vivo metabolite, deepoxy-deoxynivalenol, in broiler chickens and pigs. These species have a high exposure to these toxins, given their mainly cereal based diet. Several sample cleanup strategies were tested and further optimized by means of fractional factorial designs. A simple and straightforward sample preparation method was developed consisting out of a deproteinisation step with acetonitrile, followed by evaporation of the supernatant and reconstitution in water. The method was single laboratory validated according to European guidelines and found to be applicable for the intended purpose, with a linear response up to 200ngml(-1) and limits of quantification of 0.1-2ngml(-1). As a proof of concept, biological samples from a broiler chicken that received either deoxynivalenol, 3- or 15-acetyl-deoxynivalenol were analyzed. Preliminary results indicate nearly complete hydrolysis of 3-acetyl-deoxynivalenol to deoxynivalenol; and to a lesser extent of 15-acetyl-deoxynivalenol to deoxynivalenol. No deepoxy-deoxynivalenol was detected in any of the plasma samples. The method will be applied to study full toxicokinetic properties of deoxynivalenol, 3-acetyl-deoxynivalenol and 15-acetyl-deoxynivalenol in broiler chickens and pigs. Copyright © 2014 Elsevier B.V. All rights reserved.
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Phytotoxic effects of trichothecenes on the growth and morphology of Arabidopsis thaliana.
Masuda, Daisuke; Ishida, Mamoru; Yamaguchi, Kazuo; Yamaguchi, Isamu; Kimura, Makoto; Nishiuchi, Takumi
2007-01-01
Non-volatile sesquiterpenoids, a trichothecene family of phytotoxins such as deoxynivalenol (DON) and T-2 toxin, contain numerous molecular species and are synthesized by phytopathogenic Fusarium species. Although trichothecene chemotypes might play a role in the virulence of individual Fusarium strains, the phytotoxic action of individual trichothecenes has not been systematically studied. To perform a comparative analysis of the phytotoxic action of representative trichothecenes, the growth and morphology of Arabidopsis thaliana growing on media containing these compounds was investigated. Both DON and diacetoxyscirpenol (DAS) preferentially inhibited root elongation. DON-treated roots were less organized compared with control roots. Moreover, preferential inhibition of root growth by DON was also observed in wheat plants. In addition, T-2 toxin-treated seedlings exhibited dwarfism with aberrant morphological changes (e.g. petiole shortening, curled dark-green leaves, and reduced cell size). These results imply that the phytotoxic action of trichothecenes differed among their molecular species. Cycloheximide (CHX)-treated seedlings displayed neither feature, although it is known that trichothecenes inhibit translation in eukaryotic ribosomes. Microarray analyses suggested that T-2 toxin caused a defence response, the inactivation of brassinosteroid (BR), and the generation of reactive oxygen species in Arabidopsis. This observation is in agreement with our previous reports in which trichothecenes such as T-2 toxin have an elicitor-like activity when infiltrated into the leaves of Arabidopsis. Since it has been reported that BR plays an important role in a broad range of disease resistance in tobacco and rice, inactivation of BR might affect pathogenicity during the infection of host plants by trichothecene-producing fungi.
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Faixová, Zita; Faix, Stefan; Borutová, Radka; Leng, Lubomír
2010-09-01
This study was conducted to investigate the effects of deoxynivalenol (DON) and zearalenone (ZEA) on some biochemical indices of broiler chickens. Twenty-four Ross 308 hybrid broiler chickens of both sexes were fed diets containing maize contaminated with Fusarium mycotoxins. The diets included a control diet (DON 0.60 mg/kg feed; ZEA 0.07 mg/kg feed), an experimental 1 diet (DON 3.4 mg kg⁻¹ feed; ZEA 3.4 mg kg⁻¹ feed), and an experimental 2 diet (DON 8.2 mg kg⁻¹ feed; ZEA 8.3 mg kg⁻¹ feed). Contaminated diets were fed from 14 days of age for 14 days. Blood samples were collected from 4-week-old birds. Chicks fed a diet containing a low level of contaminated maize (experimental 1) had decreased plasma potassium, magnesium, phosphorus, total protein, albumin, triglycerides, free glycerol concentrations and increased cholesterol and calcium levels as well as alkaline phosphatase (ALP) and aspartate aminotransferase (AST) enzyme activities as compared to the control. Feeding a diet contaminated with high levels of mycotoxins (experimental 2) resulted in decreased plasma potassium, magnesium, total protein, albumin, triglycerides, free glycerol concentrations and increased plasma ALP, alanine aminotransferase (ALT) and AST enzyme activities. The effect of mycotoxin-contaminated diets on ALP activity was dose dependent. Chloride concentration was not affected by the diets. It can be concluded that feeding diets contaminated with both levels of Fusarium mycotoxins significantly affected protein, lipid and mineral metabolism as well as AST and ALP enzyme activities in broiler chickens.
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Fate of Fusarium mycotoxins in maize flour and grits during extrusion cooking.
Scudamore, Keith A; Guy, Robin C E; Kelleher, Brian; MacDonald, Susan J
2008-11-01
Extrusion technology is used widely in the manufacture of a range of breakfast cereals and snacks for human consumption and animal feeds. To minimise consumer exposure to mycotoxins, the levels of deoxynivalenol (DON) and zearalenone (ZON) in cereals/cereal products and fumonisins B(1) and B(2) (FB(1) and FB(2)) in maize are controlled by European Union legislation. Relatively few studies, however, have examined the loss of Fusarium mycotoxins during processing. The behaviour of FB(1), FB(2) and fumonisin B(3) (FB(3)), DON and ZON during extrusion of naturally contaminated maize flour and maize grits is examined using pilot-scale equipment. DON and ZON are relatively stable during extrusion cooking but the fumonisins are lost to varying degrees. There is some loss of ZON when present in low concentrations and extruded at higher moisture contents. The presence of additives, such as reducing sugars and sodium chloride, can also affect mycotoxin levels. Moisture content of the cereal feed during extrusion is important and has a greater effect than temperature, particularly on the loss of fumonisins at the lower moistures. The effects are complex and not easy to explain, although more energy input to the extruder is required for drier materials. However, on the basis of these studies, the relationship between the concentration of Fusarium toxins in the raw and finished product is toxin- and process-dependent.
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The fungal myosin I is essential for Fusarium toxisome formation.
Tang, Guangfei; Chen, Yun; Xu, Jin-Rong; Kistler, H Corby; Ma, Zhonghua
2018-01-01
Myosin-I molecular motors are proposed to function as linkers between membranes and the actin cytoskeleton in several cellular processes, but their role in the biosynthesis of fungal secondary metabolites remain elusive. Here, we found that the myosin I of Fusarium graminearum (FgMyo1), the causal agent of Fusarium head blight, plays critical roles in mycotoxin biosynthesis. Inhibition of myosin I by the small molecule phenamacril leads to marked reduction in deoxynivalenol (DON) biosynthesis. FgMyo1 also governs translation of the DON biosynthetic enzyme Tri1 by interacting with the ribosome-associated protein FgAsc1. Disruption of the ATPase activity of FgMyo1 either by the mutation E420K, down-regulation of FgMyo1 expression or deletion of FgAsc1 results in reduced Tri1 translation. The DON biosynthetic enzymes Tri1 and Tri4 are mainly localized to subcellular structures known as toxisomes in response to mycotoxin induction and the FgMyo1-interacting protein, actin, participates in toxisome formation. The actin polymerization disruptor latrunculin A inhibits toxisome assembly. Consistent with this observation, deletion of the actin-associated proteins FgPrk1 and FgEnd3 also results in reduced toxisome formation. Unexpectedly, the FgMyo1-actin cytoskeleton is not involved in biosynthesis of another secondary metabolite tested. Taken together, this study uncovers a novel function of myosin I in regulating mycotoxin biosynthesis in filamentous fungi.
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The fungal myosin I is essential for Fusarium toxisome formation
Xu, Jin-Rong
2018-01-01
Myosin-I molecular motors are proposed to function as linkers between membranes and the actin cytoskeleton in several cellular processes, but their role in the biosynthesis of fungal secondary metabolites remain elusive. Here, we found that the myosin I of Fusarium graminearum (FgMyo1), the causal agent of Fusarium head blight, plays critical roles in mycotoxin biosynthesis. Inhibition of myosin I by the small molecule phenamacril leads to marked reduction in deoxynivalenol (DON) biosynthesis. FgMyo1 also governs translation of the DON biosynthetic enzyme Tri1 by interacting with the ribosome-associated protein FgAsc1. Disruption of the ATPase activity of FgMyo1 either by the mutation E420K, down-regulation of FgMyo1 expression or deletion of FgAsc1 results in reduced Tri1 translation. The DON biosynthetic enzymes Tri1 and Tri4 are mainly localized to subcellular structures known as toxisomes in response to mycotoxin induction and the FgMyo1-interacting protein, actin, participates in toxisome formation. The actin polymerization disruptor latrunculin A inhibits toxisome assembly. Consistent with this observation, deletion of the actin-associated proteins FgPrk1 and FgEnd3 also results in reduced toxisome formation. Unexpectedly, the FgMyo1-actin cytoskeleton is not involved in biosynthesis of another secondary metabolite tested. Taken together, this study uncovers a novel function of myosin I in regulating mycotoxin biosynthesis in filamentous fungi. PMID:29357387
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Zhang, Wei; Zhang, Shihua; Zhang, Meiling; Yang, Lige; Cheng, Baojing; Li, Jianping; Shan, Anshan
2018-01-01
Deoxynivalenol (DON), zearalenone (ZEN) and fumonisin B 1 (FB 1 ) are among the most toxicologically important Fusarium toxins commonly found in nature that lead to nephrotoxicity in animals. The present study investigated that the individual and combined effects of subcytotoxic DON (0.25 μM), ZEN (20 μM) and FB 1 (10 μM) on oxidative stress and apoptosis in porcine kidney cells (PK15). In addition, the protective effect of N-acetylcysteine (NAC) against the toxicity of Fusarium toxins was also evaluated. Our results showed that the activities of glutathione reductase (GR) and total superoxide dismutase (SOD) were affected by DON, ZEN and FB 1 , and this change in activity induced reactive oxygen species (ROS) and malondialdehyde (MDA) production, increased apoptosis and regulated the mRNA expression of Bax, Bcl-2, caspase-3, caspase-9, cytochrome c (cyto c) and P53. This study demonstrated the complexity of combined mycotoxin infection since the combination of toxins exhibited more profound defects in the oxidative stress responses and apoptosis. Moreover, NAC reduced the oxidative damage and inhibited the apoptosis induced by Fusarium toxins. It was concluded that oxidative damage and apoptosis through the mitochondria-dependent channel were the mechanisms of Fusarium toxin mediated toxicity, and NAC reversed these damages to some extent. Copyright © 2017 Elsevier Ltd. All rights reserved.
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7 CFR 1421.5 - Eligible commodities.
Code of Federal Regulations, 2010 CFR
2010-01-01
...)(1) To be an eligible commodity, the commodity must be merchantable for food, feed, or other uses... poisonous to humans or animals. A commodity containing vomitoxin, aflatoxin, or Aspergillus mold may not be...
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7 CFR 1421.5 - Eligible commodities.
Code of Federal Regulations, 2011 CFR
2011-01-01
...)(1) To be an eligible commodity, the commodity must be merchantable for food, feed, or other uses... poisonous to humans or animals. A commodity containing vomitoxin, aflatoxin, or Aspergillus mold may not be...
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Tran, Anh Tuan; Kluess, Jeannette; Berk, Andreas; Paulick, Marleen; Frahm, Jana; Schatzmayr, Dian; Winkler, Janine; Kersten, Susanne; Dänicke, Sven
2018-02-01
A feeding experiment with piglets was performed to examine the efficacy of a wet preservation of Fusarium (FUS)-contaminated maize with sodium sulphite (SoS) based on deoxynivalenol (DON) and zearalenone (ZEN) residue levels in urine, bile and liquor and health traits of piglets. For this purpose, 80 castrated male piglets (7.57 ± 0.92 kg BW) were assigned to four treatment groups: CON- (control diet, with 0.09 mg DON and <0.01 mg ZEN/kg diet), CON+ (diet CON-, wet-preserved with 5 g SoS/kg maize; containing 0.05 mg DON and <0.01 mg ZEN/kg diet), FUS- (diet with mycotoxin-contaminated maize; containing 5.36 mg DON and 0.29 mg ZEN/kg diet), and FUS+ (diet FUS-, wet-preserved with 5 g SoS/kg maize; resulting in 0.83 mg DON and 0.27 mg ZEN/kg diet). After 42 d, 40 piglets (n = 10 per group) were sampled. A clear reduction of DON levels by approximately 75% was detected in all specimens of pigs fed diet FUS+. ZEN was detected in all urine, bile and liquor samples, while their metabolites were only detectable in urine and bile. Additionally, their concentrations were not influenced by SoS treatment. Among the health-related traits, feeding of FUS diets increased the total counts of leukocytes and segmented neutrophil granulocytes irrespective of SoS treatment. SoS treatment increased the total blood protein content slightly with a similar numerical trend in albumin concentration. These effects occurred at an obviously lower level in FUS-fed groups. Moreover, SoS treatment recovered the reduction of NO production induced by feeding diet FUS- indicating an effect on the redox level. As this effect only occurred in group FUS+, it is obviously related to the adverse effects of the Fusarium toxins. In conclusion, treatment of FUS-contaminated maize with SoS decreased the inner exposure with DON as indicated by the lower DON levels in various piglet specimens. However, health-related traits did not consistently reflect this decreased exposure.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Garvey, Graeme S.; McCormick, Susan P.; Alexander, Nancy J.
2009-08-14
Fusarium head blight is a devastating disease of cereal crops whose worldwide incidence is increasing and at present there is no satisfactory way of combating this pathogen or its associated toxins. There is a wide variety of trichothecene mycotoxins and they all contain a 12,13-epoxytrichothecene skeleton but differ in their substitutions. Indeed, there is considerable variation in the toxin profile across the numerous Fusarium species that has been ascribed to differences in the presence or absence of biosynthetic enzymes and their relative activity. This article addresses the source of differences in acetylation at the C15 position of the trichothecene molecule.more » Here, we present the in vitro structural and biochemical characterization of TRI3, a 15-O-trichothecene acetyltransferase isolated from F. sporotrichioides and the 'in vivo' characterization of Deltatri3 mutants of deoxynivalenol (DON) producing F. graminearum strains. A kinetic analysis shows that TRI3 is an efficient enzyme with the native substrate, 15-decalonectrin, but is inactive with either DON or nivalenol. The structure of TRI3 complexed with 15-decalonectrin provides an explanation for this specificity and shows that Tri3 and Tri101 (3-O-trichothecene acetyltransferase) are evolutionarily related. The active site residues are conserved across all sequences for TRI3 orthologs, suggesting that differences in acetylation at C15 are not due to differences in Tri3. The tri3 deletion mutant shows that acetylation at C15 is required for DON biosynthesis even though DON lacks a C15 acetyl group. The enzyme(s) responsible for deacetylation at the 15 position of the trichothecene mycotoxins have not been identified.« less
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Warth, Benedikt; Sulyok, Michael; Fruhmann, Philipp; Mikula, Hannes; Berthiller, Franz; Schuhmacher, Rainer; Hametner, Christian; Abia, Wilfred Angie; Adam, Gerhard; Fröhlich, Johannes; Krska, Rudolf
2012-07-15
Mycotoxins regularly occur in food worldwide and pose serious health risks to consumers. Since individuals can be exposed to a variety of these toxic secondary metabolites of fungi at the same time, there is a demand for proper analytical methods to assess human exposure by suitable biomarkers. This study reports on the development of a liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) method for the quantitative measurement of 15 mycotoxins and key metabolites in human urine using polarity switching. Deoxynivalenol (DON), DON-3-O-glucuronide, DON-15-O-glucuronide (D15GlcA), de-epoxy DON, nivalenol (NIV), T-2 toxin, HT-2 toxin, zearalenone, zearalenone-14-O-glucuronide, α- and β-zearalenol, fumonisins B(1) and B(2) (FB(1), FB(2)), ochratoxin A (OTA) and aflatoxin M(1) (AFM(1)) were determined without the need for any cleanup using a rapid and simple dilute and shoot approach. Validation was performed in the range of 0.005-40 µg L(-1) depending on the analyte and expected urinary concentration levels. Apparent recoveries between 78 and 119% and interday precisions of 2-17% relative standard deviation (RSD) were achieved. The applicability of the method was demonstrated by the analysis of urine samples obtained from Cameroon. In naturally contaminated urine samples up to six biomarkers of exposure (AFM(1), DON, D15GlcA, NIV, FB(1), and OTA) were detected simultaneously. We conclude that the developed LC/MS/MS method is well suited to quantify multiple mycotoxin biomarkers in human urine down to the sub-ppb range within 18 min and without any prior cleanup. The co-occurrence of several mycotoxins in the investigated samples clearly emphasizes the great potential and importance of this method to assess exposure of humans and animals to naturally occurring mycotoxins. Copyright © 2012 John Wiley & Sons, Ltd.
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Mylona, Kalliopi; Sulyok, Michael; Magan, Naresh
2012-01-01
This study examined the relationship between storage environmental factors (water activity (a(w)) (0.89-0.97) and temperature (15°C-30°C)), colonisation of wheat and maize by Fusarium graminearum and F. verticillioides respectively and the dry matter losses (DMLs) caused and quantified by contamination with deoxynivalenol (DON), zearalenone (ZEA) and fumonisins (FUMs) during storage. Fungal growth was assessed by the amount of CO(2) produced under different interacting conditions of a(w) and temperature. DMLs were quantified using the cumulative CO(2) data, and these were shown to increase as temperature and a(w) increased. The amount of DON, ZEA (wheat for human consumption) and FUMs (feed maize) produced was significantly affected by the storage conditions. The three toxins however showed different patterns of production. Optimum for DON was at the wettest conditions (0.97a(w)) and the highest temperature assessed (30°C), whereas for ZEA this shifted to 25°C. FUMs were produced in higher amounts in maize at 30°C and 0.97a(w); however, at intermediate a(w) levels (0.955a(w)), the highest production occurred at 25°C followed by 20°C. Polynomial models were developed for the effect of the storage factors on DMLs and toxin production. DMLs under different environmental conditions were significantly correlated with DON and FUMs. DON contamination was above the EU limits in at least 80% of the wheat samples with DMLs >1%, whereas at least 70% of the same samples contained ZEA above the respective EU legislative limits. Similarly, at least 75% of the maize samples with DMLs ≥ 0.9% exceeded the EU limits for the sum of FUMs in feed. These results show that it may be possible to use temporal CO(2) production during storage of grains as an indicator of the level of contamination of the grain with mycotoxins.
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Steed, A; Chandler, E; Thomsett, M; Gosman, N; Faure, S; Nicholson, P
2005-08-01
Using a set of 21 substitution lines of Triticum macha in a 'Hobbit Sib' background, it was previously demonstrated that chromosome 4A of T. macha carries significant resistance to Fusarium head blight. In the present study, the T. macha 4A resistance was further characterized in a 'Hobbit Sib' (T. macha 4A) single-recombinant chromosome doubled haploid (DH) population. Lines were phenotyped for disease resistance, yield components and deoxynivalenol (DON) mycotoxin content over two consecutive seasons. Both resistance to spread and resistance to initial infection were examined, and it was established that the resistance residing on T. macha 4A is predominantly of type I (resistance to initial infection). It was demonstrated that this type I resistance significantly lowered levels of DON accumulation in the grain and improved yield components under high disease pressure. Genotyping the DH lines using microsatellite genetic markers enabled the location of the gene(s) for resistance to be assigned to a region of the short arm of chromosome 4A, distal to microsatellite marker Xgwm601 and co-segregating with microsatellite marker Xgwm165 in this population.
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Smith, Marie-Caroline; Madec, Stéphanie; Coton, Emmanuel; Hymery, Nolwenn
2016-03-26
Some foods and feeds are often contaminated by numerous mycotoxins, but most studies have focused on the occurrence and toxicology of a single mycotoxin. Regulations throughout the world do not consider the combined effects of mycotoxins. However, several surveys have reported the natural co-occurrence of mycotoxins from all over the world. Most of the published data has concerned the major mycotoxins aflatoxins (AFs), ochratoxin A (OTA), zearalenone (ZEA), fumonisins (FUM) and trichothecenes (TCTs), especially deoxynivalenol (DON). Concerning cereals and derived cereal product samples, among the 127 mycotoxin combinations described in the literature, AFs+FUM, DON+ZEA, AFs+OTA, and FUM+ZEA are the most observed. However, only a few studies specified the number of co-occurring mycotoxins with the percentage of the co-contaminated samples, as well as the main combinations found. Studies of mycotoxin combination toxicity showed antagonist, additive or synergic effects depending on the tested species, cell model or mixture, and were not necessarily time- or dose-dependent. This review summarizes the findings on mycotoxins and their co-occurrence in various foods and feeds from all over the world as well as in vitro experimental data on their combined toxicity.
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Smith, Marie-Caroline; Madec, Stéphanie; Coton, Emmanuel; Hymery, Nolwenn
2016-01-01
Some foods and feeds are often contaminated by numerous mycotoxins, but most studies have focused on the occurrence and toxicology of a single mycotoxin. Regulations throughout the world do not consider the combined effects of mycotoxins. However, several surveys have reported the natural co-occurrence of mycotoxins from all over the world. Most of the published data has concerned the major mycotoxins aflatoxins (AFs), ochratoxin A (OTA), zearalenone (ZEA), fumonisins (FUM) and trichothecenes (TCTs), especially deoxynivalenol (DON). Concerning cereals and derived cereal product samples, among the 127 mycotoxin combinations described in the literature, AFs+FUM, DON+ZEA, AFs+OTA, and FUM+ZEA are the most observed. However, only a few studies specified the number of co-occurring mycotoxins with the percentage of the co-contaminated samples, as well as the main combinations found. Studies of mycotoxin combination toxicity showed antagonist, additive or synergic effects depending on the tested species, cell model or mixture, and were not necessarily time- or dose-dependent. This review summarizes the findings on mycotoxins and their co-occurrence in various foods and feeds from all over the world as well as in vitro experimental data on their combined toxicity. PMID:27023609
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KONGKAPAN, Jutamart; POAPOLATHEP, Saranya; ISARIYODOM, Supaporn; KUMAGAI, Susumu; POAPOLATHEP, Amnart
2015-01-01
Mycotoxins are secondary fungal metabolites that are typically present in grain and feed ingredients used for animal feeds. An analytical method using LC-ESI-MS/MS was developed to quantify nine mycotoxins, consisting of aflatoxin B1 (AFB1), AFB2, AFG1, AFG2, T-2 toxin, deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEA) and ochratoxin A (OTA) in broiler feeds. In total, 100 samples of broiler feeds were collected from poultry farms in Central Thailand. The survey found that AFB1 and ZEA were the most prevalent mycotoxins in the feed samples at percentages of 93% and 63%, respectively. The limit of detections (LODs) of investigated mycotoxins was 0.20–0.78 ng/g. AFB2, DON, AFG1, NIV and T-2 toxin were also detectable at low contamination levels with percentages of 20%, 9%, 7%, 5% and 1%, respectively, whereas OTA and AFG2 were not detected in any of the feed samples. These results suggest that there is a very low level of risk of the exposure to mycotoxins in feeds obtained from broiler farms in Central Thailand. PMID:26477362
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Blandino, Massimo; Sovrani, Valentina; Marinaccio, Federico; Reyneri, Amedeo; Rolle, Luca; Giacosa, Simone; Locatelli, Monica; Bordiga, Matteo; Travaglia, Fabiano; Coïsson, Jean Daniel; Arlorio, Marco
2013-12-01
A strategy to maximise the health benefits of wheat-based products enriched with refined flour and selected fractions of kernel, obtained by sequential pearling, has been tested. Five mixtures of refined commercial flour with an increasing replacement of a pearled wheat fraction were used to prepare bread and were compared with a control for the dough rheological properties (Mixolab® parameters), the bioactive compound content, deoxynivalenol (DON) contamination and the physical properties (volume, crust colour, instrumental crunchiness and crumb texture profile analysis parameters). The antioxidant and dietary fibre contents increased linearly as the flour was enriched with the pearled fraction. The dietary fibre, β-glucan, total phenolic, alkylresorcinol content and the antioxidant activity increased significantly at a replacement level of 10%, while the technological properties were not significantly different from those of the control. It has been shown that refined flour could be enriched through the addition of a selected wheat pearled fraction and the bioactive compound content of composite bread could be improved, while few rheological and technological differences could be obtained and the risk for DON contamination could be limited. Copyright © 2013 Elsevier Ltd. All rights reserved.
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Mitochondrial functions of THP-1 monocytes following the exposure to selected natural compounds.
Schultze, Nadin; Wanka, Heike; Zwicker, Paula; Lindequist, Ulrike; Haertel, Beate
2017-02-15
The immune system is an important target of various xenobiotics, which may lead to severe adverse effects including immunosuppression or inappropriate immunostimulation. Mitochondrial toxicity is one possibility by which xenobiotics exert their toxic effects in cells or organs. In this study, we investigated the impact of three natural compounds, cyclosporine A (CsA), deoxynivalenol (DON) and cannabidiol (CBD) on mitochondrial functions in the THP-1 monocytic cell line. The cells were exposed for 24h to two different concentrations (IC 10 and IC 50 determined by MTT) of each compound. The cells showed concentration-dependent elevated intracellular reactive oxygen species (iROS) and induction of apoptosis (except DON) in response to the three test compounds. Mitochondrial functions were characterized by using bioenergetics profiling experiments. In THP-1 monocytes, the IC 50 of CsA decreased basal and maximal respiration as well as ATP production with an impact on spare capacity indicating a mitochondrial dysfunction. Similar reaction patterns were observed following CBD exposure. The basal respiration level and ATP-production decreased in the THP-1 cells exposed to the IC 50 of DON with no major impact on mitochondrial function. In conclusion, impaired mitochondrial function was accompanied by elevated iROS and apoptosis level in a monocytic cell line exposed to CsA and CBD. Mitochondrial dysfunction may be one explanation for the cytotoxicity of CBD and CsA also in other in immune cells. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
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Multiplexed detection of mycotoxins in foods with a regenerable array.
Ngundi, Miriam M; Shriver-Lake, Lisa C; Moore, Martin H; Ligler, Frances S; Taitt, Chris R
2006-12-01
The occurrence of different mycotoxins in cereal products calls for the development of a rapid, sensitive, and reliable detection method that is capable of analyzing samples for multiple toxins simultaneously. In this study, we report the development and application of a multiplexed competitive assay for the simultaneous detection of ochratoxin A (OTA) and deoxynivalenol (DON) in spiked barley, cornmeal, and wheat, as well as in naturally contaminated maize samples. Fluoroimmunoassays were performed with the Naval Research Laboratory array biosensor, by both a manual and an automated version of the system. This system employs evanescent-wave fluorescence excitation to probe binding events as they occur on the surface of a waveguide. Methanolic extracts of the samples were diluted threefold with buffer containing a mixture of fluorescent antibodies and were then passed over the arrays of mycotoxins immobilized on a waveguide. Fluorescent signals of the surface-bound antibody-antigen complexes decreased with increasing concentrations of free mycotoxins in the extract. After sample analysis was completed, surfaces were regenerated with 6 M guanidine hydrochloride in 50 mM glycine, pH 2.0. The limits of detection determined by the manual biosensor system were as follows: 1, 180, and 65 ng/g for DON and 1, 60, and 85 ng/g for OTA in cornmeal, wheat, and barley, respectively. The limits of detection in cornmeal determined with the automated array biosensor were 15 and 150 ng/g for OTA and DON, respectively.
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Identifying Rare FHB-Resistant Segregants in Intransigent Backcross and F2 Winter Wheat Populations.
Clark, Anthony J; Sarti-Dvorjak, Daniela; Brown-Guedira, Gina; Dong, Yanhong; Baik, Byung-Kee; Van Sanford, David A
2016-01-01
Fusarium head blight (FHB), caused mainly by Fusarium graminearum Schwabe [telomorph: Gibberella zeae Schwein.(Petch)] in the US, is one of the most destructive diseases of wheat (Triticum aestivum L. and T. durum L.). Infected grain is usually contaminated with deoxynivalenol (DON), a serious mycotoxin. The challenge in FHB resistance breeding is combining resistance with superior agronomic and quality characteristics. Exotic QTL are widely used to improve FHB resistance. Success depends on the genetic background into which the QTL are introgressed, whether through backcrossing or forward crossing; QTL expression is impossible to predict. In this study four high-yielding soft red winter wheat breeding lines with little or no scab resistance were each crossed to a donor parent (VA01W-476) with resistance alleles at two QTL: Fhb1 (chromosome 3BS) and QFhs.nau-2DL (chromosome 2DL) to generate backcross and F2 progeny. F2 individuals were genotyped and assigned to 4 groups according to presence/ absence of resistance alleles at one or both QTL. The effectiveness of these QTL in reducing FHB rating, incidence, index, severity, Fusarium-damaged kernels (FDK) and DON, in F2-derived lines was assessed over 2 years. Fhb1 showed an average reduction in DON of 17.5%, and conferred significant resistance in 3 of 4 populations. QFhs.nau-2DL reduced DON 6.7% on average and conferred significant resistance in 2 of 4 populations. The combination of Fhb1 and QFhs.nau-2DL resistance reduced DON 25.5% across all populations. Double resistant lines had significantly reduced DON compared to double susceptible lines in 3 populations. Backcross derived progeny were planted in replicated yield trials (2011 and 2012) and in a scab nursery in 2012. Several top yielding lines performed well in the scab nursery, with acceptable DON concentrations, even though the average effect of either QTL in this population was not significant. Population selection is often viewed as an "all or nothing" process: if the average resistance level is insufficient, the population is discarded. These results indicate that it may be possible to find rare segregants which combine scab resistance, superior agronomic performance and acceptable quality even in populations in which the average effect of the QTL is muted or negligible.
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Mesterházy, Ákos; Tóth, Beáta; Varga, Monika; Bartók, Tibor; Szabó-Hevér, Ágnes; Farády, László; Lehoczki-Krsjak, Szabolcs
2011-01-01
Fungicide application is a key factor in the control of mycotoxin contamination in the harvested wheat grain. However, the practical results are often disappointing. In 2000-2004, 2006-2008 and 2007 and 2008, three experiments were made to test the efficacy of fungicide control on Fusarium Head Blight (FHB) in wheat and to find ways to improve control of the disease and toxin contamination. In a testing system we have used for 20 years, tebuconazole and tebuconazole + prothioconazole fungicides regularly reduced symptoms by about 80% with a correlating reduction in toxin contamination. Averages across the years normally show a correlation of r = 0.90 or higher. The stability differences (measured by the stability index) between the poorest and the best fungicides are about 10 or more times, differing slightly in mycotoxin accumulation, FHB index (severity) and Fusarium damaged kernels (FDK). The weak fungicides, like carbendazim, were effective only when no epidemic occurred or epidemic severity was at a very low level. Similar fungicide effects were seen on wheat cultivars which varied in FHB resistance. In this study, we found three fold differences in susceptibility to FHB between highly susceptible and moderately resistant cultivars when treated with fungicides. In the moderately resistant cultivars, about 50% of the fungicide treatments lowered the DON level below the regulatory limit. In the most susceptible cultivars, all fungicides failed to reduce mycotoxin levels low enough for grain acceptance, in spite of the fact that disease was significantly reduced. The results correlated well with the results of the large-scale field tests of fungicide application at the time of natural infection. The Turbo FloodJet nozzle reduced FHB incidence and DON contamination when compared to the TeeJet XR nozzle. Overall, the data suggest that significant decreases in FHB incidence and deoxynivalenol contamination in field situations are possible with proper fungicide applications. Additionally, small plot tests can be used to evaluate the quality of the field disease and toxin production. PMID:22174980
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Moon, Yuseok; Yang, Hyun; Park, Seung-Hwan
Mucosal epithelium senses external toxic insults and transmits the danger signals into the epithelial cells in order to activate a broad range of inflammatory responses. However, pre-exposure to the commensal endotoxins can induce inflammatory tolerance and maintain the homeostasis without excessive immune responses. We recently reported that ribotoxin deoxynivalenol (DON) and its derivatives elicited the pro-inflammatory response as the mucosal insults in human epithelial cells. Taking the knowledge into consideration, we tested the hypothesis that endotoxin pre-exposure can attenuate ribotoxin-induced epithelial interleukin-8 (IL-8) production via a tolerance mechanism. Pre-exposure to endotoxin repressed IL-8 release and its gene expression. However, inflammatorymore » tolerance was not mediated by the attenuated NF-{kappa}B activation which has been generally recognized as the major mediator of LPS-mediated toll-like receptor (TLR) signaling pathway. Instead, pre-exposure to endotoxin was observed to trigger the delayed induction of peroxisome proliferator-activated receptor gamma (PPAR-{gamma}) which contributed to the diminished IL-8 production in the human epithelial cells. Moreover, endogenous PPAR-{gamma} agonist suppressed toxicant-mediated interleukin-8 production and IL-8 mRNA stability. Taken together, endotoxin induced hypo-production of pro-inflammatory cytokine IL-8 in the human epithelial cells, which was associated with the delayed activation of PPAR-{gamma} expression by pre-existing endotoxin.« less
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Li, Chenglong; Wen, Kai; Mi, Tiejun; Zhang, Xiya; Zhang, Huiyan; Zhang, Suxia; Shen, Jianzhong; Wang, Zhanhui
2016-05-15
Multi-analyte immunoassays have attracted increasing attention due to their short assay times, low sample consumption, and reduced detection costs per assay. In this work, we describe a homologous and high-throughput multi-wavelength fluorescence polarization immunoassay (MWFPIA) for the multiplexed detection of mycotoxins. Three typical Fusarium mycotoxins, deoxynivalenol (DON), T-2 toxin and fumonisin B1 (FB1), were labeled with different dyes. Tracers and specific monoclonal antibodies (mAbs) were employed in the MWFPIA to simultaneously detect the three mycotoxins. Under optimal conditions, the limits of detection using MWFPIA were 242.0 μg kg(-1) for DON, 17.8 μg kg(-1) for T-2 toxin and 331.5 μg kg(-1) for FB1, providing sufficient sensitivity to meet the action levels of these three contaminants in maize as set by the European Union. The use of a methanol/water (2:3, v/v) mixture for sample pretreatment allowed recoveries ranging from 76.5-106.3%, with coefficients of variation less than 21.7%. The total time of analysis, including sample preparation, was less than 30 min. Twenty naturally contaminated maize samples were tested using MWFPIA and HPLC-MS/MS, with correlation coefficients (R(2)) of 0.97 for DON and 0.99 for FB1. By changing the targets of interest, homologous MWFPIA, a method with high sensitivity, a simple procedure and a short analysis time, can easily be extended to other chemical contaminants. Thus, MWFPIA represents a versatile strategy for food safety analysis. Copyright © 2015 Elsevier B.V. All rights reserved.
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Bencze, Szilvia; Puskás, Katalin; Vida, Gyula; Karsai, Ildikó; Balla, Krisztina; Komáromi, Judit; Veisz, Ottó
2017-08-01
Increasing atmospheric CO 2 concentration not only has a direct impact on plants but also affects plant-pathogen interactions. Due to economic and health-related problems, special concern was given thus in the present work to the effect of elevated CO 2 (750 μmol mol -1 ) level on the Fusarium culmorum infection and mycotoxin contamination of wheat. Despite the fact that disease severity was found to be not or little affected by elevated CO 2 in most varieties, as the spread of Fusarium increased only in one variety, spike grain number and/or grain weight decreased significantly at elevated CO 2 in all the varieties, indicating that Fusarium infection generally had a more dramatic impact on the grain yield at elevated CO 2 than at the ambient level. Likewise, grain deoxynivalenol (DON) content was usually considerably higher at elevated CO 2 than at the ambient level in the single-floret inoculation treatment, suggesting that the toxin content is not in direct relation to the level of Fusarium infection. In the whole-spike inoculation, DON production did not change, decreased or increased depending on the variety × experiment interaction. Cooler (18 °C) conditions delayed rachis penetration while 20 °C maximum temperature caused striking increases in the mycotoxin contents, resulting in extremely high DON values and also in a dramatic triggering of the grain zearalenone contamination at elevated CO 2 . The results indicate that future environmental conditions, such as rising CO 2 levels, may increase the threat of grain mycotoxin contamination.
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Kos, Gregor; Krska, Rudolf; Lohninger, Hans; Griffiths, Peter R
2004-01-01
An investigation into the rapid detection of mycotoxin-producing fungi on corn by two mid-infrared spectroscopic techniques was undertaken. Corn samples from a single genotype (RWA2, blanks, and contaminated with Fusarium graminearum) were ground, sieved and, after appropriate sample preparation, subjected to mid-infrared spectroscopy using two different accessories (diffuse reflection and attenuated total reflection). The measured spectra were evaluated with principal component analysis (PCA) and the blank and contaminated samples were classified by cluster analysis. Reference data for fungal metabolites were obtained with conventional methods. After extraction and clean-up, each sample was analyzed for the toxin deoxynivalenol (DON) by gas chromatography with electron capture detection (GC-ECD) and ergosterol (a parameter for the total fungal biomass) by high-performance liquid chromatography with diode array detection (HPLC-DAD). The concentration ranges for contaminated samples were 880-3600 microg/kg for ergosterol and 300-2600 microg/kg for DON. Classification efficiency was 100% for ATR spectra. DR spectra did not show as obvious a clustering of contaminated and blank samples. Results and trends were also observed in single spectra plots. Quantification using a PLS1 regression algorithm showed good correlation with DON reference data, but a rather high standard error of prediction (SEP) with 600 microg/kg (DR) and 490 microg/kg (ATR), respectively, for ergosterol. Comparing measurement procedures and results showed advantages for the ATR technique, mainly owing to its ease of use and the easier interpretation of results that were better with respect to classification and quantification.
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Functional analyses of heterotrimeric G protein Gα and Gβ subunits in Gibberella zeae
Yu, Hye-Young; Seo, Jeong-Ah; Kim, Jung-Eun; Han, Kap-Hoon; Shim, Won-Bo; Yun, Sung-Hwan; Lee, Yin-Won
2008-01-01
The homothallic ascomycete fungus Gibberella zeae (anamorph: Fusarium graminearum) is a major toxigenic plant pathogen that causes head blight disease on small-grain cereals. The fungus produces the mycotoxins deoxynivalenol (DON) and zearalenone (ZEA) in infected hosts, posing a threat to human and animal health. Despite its agricultural and toxicological importance, the molecular mechanisms underlying its growth, development and virulence remain largely unknown. To better understand such mechanisms, we studied the heterotrimeric G proteins of G. zeae, which are known to control crucial signalling pathways that regulate various cellular and developmental responses in fungi. Three putative Gα subunits, GzGPA1, GzGPA2 and GzGPA3, and one Gβ subunit, GzGPB1, were identified in the F. graminearum genome. Deletion of GzGPA1, a homologue of the Aspergillus nidulans Gα gene fadA, resulted in female sterility and enhanced DON and ZEA production, suggesting that GzGPA1 is required for normal sexual reproduction and repression of toxin biosynthesis. The production of DON and ZEA was also enhanced in the GzGPB1 mutant, suggesting that both Gα GzGPA1 and Gβ GzGPB1 negatively control mycotoxin production. Deletion of GzGPA2, which encodes a Gα protein similar to A. nidulans GanB, caused reduced pathogenicity and increased chitin accumulation in the cell wall, implying that GzGPA2 has multiple functions. Our study shows that G. zeae heterotrimeric G protein subunits can regulate vegetative growth, sexual development, toxin production and pathogenicity. PMID:18227243
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Forrer, Hans-Rudolf; Musa, Tomke; Schwab, Fabienne; Jenny, Eveline; Bucheli, Thomas D; Wettstein, Felix E; Vogelgsang, Susanne
2014-02-26
Suspensions or solutions with 1% of Chinese galls (Galla chinensis, GC) or 1% of tannic acid (TA), inhibited germination of conidia or mycelium growth of Fusarium graminearum (FG) by 98%-100% or by 75%-80%, respectively, whereas dried bark from buckthorn (Frangula alnus, FA) showed no effect at this concentration. In climate chamber experiments where the wheat variety "Apogee" was artificially inoculated with FG and F. crookwellense (FCr) and treated with 5% suspensions of TA, GC and FA, the deoxynivalenol (DON) content in grains was reduced by 81%, 67% and 33%, respectively. In field experiments with two commercial wheat varieties and artificial or semi-natural inoculations, mean DON reductions of 66% (TA) and 58% (FA), respectively, were obtained. Antifungal toxicity can explain the high efficacies of TA and GC but not those of FA. The Fusarium head blight (FHB) and mycotoxin reducing effect of FA is probably due to elicitation of resistance in wheat plants. With semi-natural inoculation, a single FA application in the first half of the flowering period performed best. However, we assume that applications of FA at the end of ear emergence and a treatment, triggered by an infection period, with TA or GC during flowering, might perform better than synthetic fungicides.
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Forrer, Hans-Rudolf; Musa, Tomke; Schwab, Fabienne; Jenny, Eveline; Bucheli, Thomas D.; Wettstein, Felix E.; Vogelgsang, Susanne
2014-01-01
Suspensions or solutions with 1% of Chinese galls (Galla chinensis, GC) or 1% of tannic acid (TA), inhibited germination of conidia or mycelium growth of Fusarium graminearum (FG) by 98%–100% or by 75%–80%, respectively, whereas dried bark from buckthorn (Frangula alnus, FA) showed no effect at this concentration. In climate chamber experiments where the wheat variety “Apogee” was artificially inoculated with FG and F. crookwellense (FCr) and treated with 5% suspensions of TA, GC and FA, the deoxynivalenol (DON) content in grains was reduced by 81%, 67% and 33%, respectively. In field experiments with two commercial wheat varieties and artificial or semi-natural inoculations, mean DON reductions of 66% (TA) and 58% (FA), respectively, were obtained. Antifungal toxicity can explain the high efficacies of TA and GC but not those of FA. The Fusarium head blight (FHB) and mycotoxin reducing effect of FA is probably due to elicitation of resistance in wheat plants. With semi-natural inoculation, a single FA application in the first half of the flowering period performed best. However, we assume that applications of FA at the end of ear emergence and a treatment, triggered by an infection period, with TA or GC during flowering, might perform better than synthetic fungicides. PMID:24577585
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Mycotoxin-Containing Diet Causes Oxidative Stress in the Mouse
Hou, Yan-Jun; Zhao, Yong-Yan; Xiong, Bo; Cui, Xiang-Shun; Kim, Nam-Hyung; Xu, Yin-Xue; Sun, Shao-Chen
2013-01-01
Mycotoxins which mainly consist of Aflatoxin (AF), Zearalenone (ZEN) and Deoxynivalenol (DON) are commonly found in many food commodities. Although each component has been shown to cause liver toxicity and oxidative stress in several species, there is no evidence regarding the effect of naturally contained multiple mycotoxins on tissue toxicity and oxidative stress in vivo. In the present study, mycotoxins-contaminated maize (AF 597 µg/kg, ZEN 729 µg/kg, DON 3.1 mg/kg maize) was incorporated into the diet at three different doses (0, 5 and 20%) to feed the mice, and blood and tissue samples were collected to examine the oxidative stress related indexes. The results showed that the indexes of liver, kidney and spleen were all increased and the liver and kidney morphologies changed in the mycotoxin-treated mice. Also, the treatment resulted in the elevated glutathione peroxidase (GPx) activity and malondialdehyde (MDA) level in the serum and liver, indicating the presence of the oxidative stress. Moreover, the decrease of catalase (CAT) activity in the serum, liver and kidney as well as superoxide dismutase (SOD) activity in the liver and kidney tissue further confirmed the occurrence of oxidative stress. In conclusion, our data indicate that the naturally contained mycotoxins are toxic in vivo and able to induce the oxidant stress in the mouse. PMID:23555961
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Valle-Algarra, F M; Mateo, E M; Medina, A; Mateo, F; Gimeno-Adelantado, J V; Jimenez, M
2009-06-01
Ochratoxin A (OTA) and type B trichothecenes are mycotoxins that occur frequently in cereals and thus can be found in cereal by-products such as bread. The aim of this work was to study the variation of the levels of OTA, deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON) and nivalenol (NIV) during the bread-making process. This was done by using wheat flour spiked with different levels of toxins. Mycotoxin levels were controlled after fermentation of the dough with yeasts (Saccharomyces cerevisiae) and after further baking at different temperature-time combinations. Analysis of variance (ANOVA) of the results showed a significant reduction in OTA level (p < 0.05) during fermentation of the dough. The reduction ranged between 29.8% and 33.5%, depending on the initial concentration of toxin in the flour. During this period, the level of the other mycotoxins studied was not modified. By contrast, in the baking phase there were significant changes in the levels of the four mycotoxins, although the reduction was similar under all the baking conditions. Considering all the temperature-time conditions tested, it can be concluded that during the baking period the average reduction of OTA, NIV, 3-ADON, and DON was 32.9%, 76.9%, 65.6%, and 47.9%, respectively.
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Bönnighausen, Jakob; Gebhard, Daniel; Kröger, Cathrin; Hadeler, Birgit; Tumforde, Thomas; Lieberei, Reinhard; Bergemann, Jörg; Schäfer, Wilhelm; Bormann, Jörg
2015-12-01
The cereal pathogen Fusarium graminearum threatens food and feed production worldwide. It reduces the yield and poisons the remaining kernels with mycotoxins, notably deoxynivalenol (DON). We analyzed the importance of gamma-aminobutanoic acid (GABA) metabolism for the life cycle of this fungal pathogen. GABA metabolism in F. graminearum is partially regulated by the global nitrogen regulator AreA. Genetic disruption of the GABA shunt by deletion of two GABA transaminases renders the pathogen unable to utilize the plant stress metabolites GABA and putrescine. The mutants showed increased sensitivity against oxidative stress, GABA accumulation in the mycelium, downregulation of two key enzymes of the TCA cycle, disturbed potential gradient in the mitochondrial membrane and lower mitochondrial oxygen consumption. In contrast, addition of GABA to the wild type resulted in its rapid turnover and increased mitochondrial steady state oxygen consumption. GABA concentrations are highly upregulated in infected wheat tissues. We conclude that GABA is metabolized by the pathogen during infection increasing its energy production, whereas the mutants accumulate GABA intracellularly resulting in decreased energy production. Consequently, the GABA mutants are strongly reduced in virulence but, because of their DON production, are able to cross the rachis node. © 2015 John Wiley & Sons Ltd.
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Kos, Gregor; Lohninger, Hans; Krska, Rudolf
2003-03-01
A novel method, which enables the determination of fungal infection with Fusarium graminearum on corn within minutes, is presented. The ground sample was sieved and the particle size fraction between >250 and 100 microm was used for mid-infrared/attenuated total reflection (ATR) measurements. The sample was pressed onto the ATR crystal, and reproducible pressure was applied. After the spectra were recorded, they were subjected to principle component analysis (PCA) and classified using cluster analysis. Observed changes in the spectra reflected changes in protein, carbohydrate, and lipid contents. Ergosterol (for the total fungal biomass) and the toxin deoxynivalenol (DON; a secondary metabolite) of Fusarium fungi served as reference parameters, because of their relevance for the examination of corn based food and feed. The repeatability was highly improved by sieving prior to recording the spectra, resulting in a better clustering in PCA score/score plots. The developed method enabled the separation of samples with a toxin content of as low as 310 microg/kg from noncontaminated (blank) samples. Investigated concentration ranges were 880-3600 microg/kg for ergosterol and 310-2596 microg/kg for DON. The percentage of correctly classified samples was up to 100% for individual samples compared with a number of blank samples.
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Chen, Quansheng; Hu, Weiwei; Sun, Cuicui; Li, Huanhuan; Ouyang, Qin
2016-09-28
Rare earth-doped upconversion nanoparticles (UCNPs) have promising potentials in biodetection due to their unique frequency upconverting capability and high detection sensitivity. This paper reports an improved UCNPs-based fluorescence probe for dual-sensing of Aflatoxin B1 (AFB1) and Deoxynivalenol (DON) using a magnetism-induced separation and the specific formation of antibody-targets complex. Herein, the improved UCNPs, which were namely NaYF4:Yb/Ho/Gd and NaYF4:Yb/Tm/Gd, were systematically studied based on the optimization of reaction time, temperature and the concentration of dopant ions with simultaneous phase and size controlled NaYF4 nanoparticles; and the targets were detected using the pattern of competitive combination assay. Under an optimized condition, the advanced fluorescent probes revealed stronger fluorescent properties, broader biological applications and better storage stabilities compared to traditional UCNPs-based ones; and ultrasensitive determinations of AFB1 and DON were achieved under a wide sensing range of 0.001-0.1 ng ml(-1) with the limit of detection (LOD) of 0.001 ng ml(-1). Additionally, the applicability of the improved nanosensor for the detection of mycotoxins was also confirmed in adulterated oil samples. Copyright © 2016 Elsevier B.V. All rights reserved.
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Machado, Jessica M D; Soares, Ruben R G; Chu, Virginia; Conde, João P
2018-01-15
The field of microfluidics holds great promise for the development of simple and portable lab-on-a-chip systems. The use of capillarity as a means of fluidic manipulation in lab-on-a-chip systems can potentially reduce the complexity of the instrumentation and allow the development of user-friendly devices for point-of-need analyses. In this work, a PDMS microchannel-based, colorimetric, autonomous capillary chip provides a multiplexed and semi-quantitative immunodetection assay. Results are acquired using a standard smartphone camera and analyzed with a simple gray scale quantification procedure. The performance of this device was tested for the simultaneous detection of the mycotoxins ochratoxin A (OTA), aflatoxin B1 (AFB1) and deoxynivalenol (DON) which are strictly regulated food contaminants with severe detrimental effects on human and animal health. The multiplexed assay was performed approximately within 10min and the achieved sensitivities of<40, 0.1-0.2 and<10ng/mL for OTA, AFB1 and DON, respectively, fall within the majority of currently enforced regulatory and/or recommended limits. Furthermore, to assess the potential of the device to analyze real samples, the immunoassay was successfully validated for these 3 mycotoxins in a corn-based feed sample after a simple sample preparation procedure. Copyright © 2017 Elsevier B.V. All rights reserved.
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Profitability of Integrated Management of Fusarium Head Blight in North Carolina Winter Wheat.
Cowger, Christina; Weisz, Randy; Arellano, Consuelo; Murphy, Paul
2016-08-01
Fusarium head blight (FHB) is one of the most difficult small-grain diseases to manage, due to the partial effectiveness of management techniques and the narrow window of time in which to apply fungicides profitably. The most effective management approach is to integrate cultivar resistance with FHB-specific fungicide applications; yet, when forecasted risk is intermediate, it is often unclear whether such an application will be profitable. To model the profitability of FHB management under varying conditions, we conducted a 2-year split-plot field experiment having as main plots high-yielding soft red winter wheat cultivars, four moderately resistant (MR) and three susceptible (S) to FHB. Subplots were sprayed at flowering with Prosaro or Caramba, or left untreated. The experiment was planted in seven North Carolina environments (location-year combinations); three were irrigated to promote FHB development and four were not irrigated. Response variables were yield, test weight, disease incidence, disease severity, deoxynivalenol (DON), Fusarium-damaged kernels, and percent infected kernels. Partial profits were compared in two ways: first, across low-, medium-, or high-DON environments; and second, across environment-cultivar combinations divided by risk forecast into "do spray" and "do not spray" categories. After surveying DON and test weight dockage among 21 North Carolina wheat purchasers, three typical market scenarios were used for modeling profitability: feed-wheat, flexible (feed or flour), and the flour market. A major finding was that, on average, MR cultivars were at least as profitable as S cultivars, regardless of epidemic severity or market. Fungicides were profitable in the feed-grain and flexible markets when DON was high, with MR cultivars in the flexible or flour markets when DON was intermediate, and on S cultivars aimed at the flexible market. The flour market was only profitable when FHB was present if DON levels were intermediate and cultivar resistance was combined with a fungicide. It proved impossible to use the risk forecast to predict profitability of fungicide application. Overall, the results indicated that cultivar resistance to FHB was important for profitability, an FHB-targeted fungicide expanded market options when risk was moderate or high, and the efficacy of fungicide decision-making is reduced by factors that limit the accuracy of risk forecasts.
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Kouadio, James Halbin; Lattanzio, Veronica M T; Ouattara, Djeneba; Kouakou, Brou; Visconti, Angelo
2014-01-01
The aim of the presented study was to investigate the mycotoxin exposure of Ivorian population related to the consumption patterns of maize, peanuts, millet, and cassava product (attiéké). Maize flour samples (n = 51) were purchased from all Abidjan local markets, in the south of Ivory Coast, and urine (n = 99) was collected during the same reference period (July-September 2011) from volunteers living in Abidjan and Daloa cities. Reversed-phase liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (LC-ESI-MS/MS) was used to analyze aflatoxins (AFB1, AFB2, AFG1, and AFG2), ochratoxin A (OTA), fumonisins (FB1, FB2), deoxynivalenol (DON), zearalenone (ZEA), and T-2 and HT-2 toxins in maize flour samples, and their relevant biomarkers (AFM1, DON, DON + de-epoxydeoxynivalenol (DOM-1), FB1, α-zearalenol (ZOL), β-ZOL, and OTA) in urine samples. Critical maize contamination was observed by AFs occurrence (total AFs 4.5 - 330.0 μg/kg) while OTA was found in 13% of samples analyzed. AFM1 was detected in 40% of urines samples (0.06 - 14.11 ng/ml), OTA in 37% (0.01 - 0.42 ng/ml), FB1 in 27% (0.07 to 15.31 ng/ml) and, DON was found in 21% of samples at levels up to 10.0 ng/ml. The correlation coefficients (R(2)) obtained by plotting the percentage of biomarker occurrence (positive samples) versus the frequency of food consumption revealed maize, peanuts, millet and attiéké were strongly linked to AFB1 and OTA exposure with values of R(2) ranged from 0.462 to 0.956. The present study provided data on mycotoxin risk in Ivory Coast, revealing a frequent co-exposure to the major mycotoxins such as AFs, OTA, and fumonisins, which appeared to be related to the frequency of peanuts, maize, millet and attiéké consumption.
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Sissener, Nini H; Hemre, Gro-Ingunn; Lall, Santosh P; Sagstad, Anita; Petersen, Kjell; Williams, Jason; Rohloff, Jens; Sanden, Monica
2011-07-01
The present study was conducted to follow up on apparent differences in growth, relative organ sizes, cellular stress and immune function in Atlantic salmon fed feed containing GM Bacillus thuringiensis maize compared with feed containing the non-modified parental maize line. Gene expression profiling on the distal intestinal segment and liver was performed by microarray, and selected genes were followed up by quantitative PCR (qPCR). In the liver, qPCR revealed some differentially regulated genes, including up-regulation of gelsolin precursor, down-regulation of ferritin heavy subunit and a tendency towards down-regulation of metallothionein (MT)-B. This, combined with the up-regulation of anti-apoptotic protein NR13 and similar tendencies for ferritin heavy chain and MT-A and -B in the distal intestine, suggests changes in cellular stress/antioxidant status. This corresponds well with and strengthens previous findings in these fish. To exclude possible confounding factors, the maize ingredients were analysed for mycotoxins and metabolites. The GM maize contained 90 μg/kg of deoxynivalenol (DON), while the non-GM maize was below the detection limit. Differences were also observed in the metabolite profiles of the two maize varieties, some of which seemed connected to the mycotoxin level. The effects on salmon observed in the present and previous studies correspond relatively well with the effects of DON as reported in the literature for other production animals, but knowledge regarding effects and harmful dose levels in fish is scarce. Thus, it is difficult to conclude whether the observed effects are caused by the DON level or by some other aspect of the GM maize ingredient.
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Akbari, Peyman; Fink-Gremmels, Johanna; Willems, Rianne H A M; Difilippo, Elisabetta; Schols, Henk A; Schoterman, Margriet H C; Garssen, Johan; Braber, Saskia
2017-08-01
The direct effects of galacto-oligosaccharides (GOS), including Vivinal ® GOS syrup (VGOS) and purified Vivinal ® GOS (PGOS), on the epithelial integrity and corresponding interleukin-8 (IL-8/CXCL8) release were examined in a Caco-2 cell model for intestinal barrier dysfunction. To investigate structure-activity relationships, the effects of individual DP fractions of VGOS were evaluated. Moreover, the obtained results with GOS were compared with Caco-2 monolayers incubated with fructo-oligosaccharides (FOS) and inulin. Caco-2 monolayers were pretreated (24 h) with or without specific oligosaccharides or DP fractions of VGOS (DP2 to DP6) before being exposed for 12 or 24 h to the fungal toxin deoxynivalenol (DON). Transepithelial electrical resistance and lucifer yellow permeability were measured to investigate barrier integrity. A calcium switch assay was used to study the reassembly of tight junction proteins. Release of CXCL8, a typical marker for inflammation, was quantified by ELISA. In comparison with PGOS, FOS and inulin, VGOS showed the most pronounced protective effect on the DON-induced impairment of the monolayer integrity, acceleration of the tight junction reassembly and the subsequent CXCL8 release. DP2 and DP3 in concentrations occurring in VGOS prevented the DON-induced epithelial barrier disruption, which could be related to their high prevalence in VGOS. However, no effects of the separate DP GOS fractions were observed on CXCL8 release. This comparative study demonstrates the direct, microbiota-independent effects of oligosaccharides on the intestinal barrier function and shows the differences between individual galacto- and fructo-oligosaccharides. This microbiota-independent effect of oligosaccharides depends on the oligosaccharide structure, DP length and concentration.
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Effects of feed-borne Fusarium mycotoxins on hematology and immunology of laying hens.
Chowdhury, S R; Smith, T K; Boermans, H J; Woodward, B
2005-12-01
Feeding grains naturally contaminated with Fusarium mycotoxins has been shown to alter metabolism and performance of laying hens. The objectives of the current experiment were to examine the effects of feeding grains naturally contaminated with Fusarium mycotoxins on hematology and immunological indices and functions of laying hens and the possible protective effect of feeding a polymeric glucomannan mycotoxin adsorbent (GMA). One hundred forty-four laying hens were fed for 12 wk with diets formulated with (1) uncontaminated grains, (2) contaminated grains, or (3) contaminated grains + 0.2% GMA. Fusarium mycotoxins such as deoxynivalenol (DON, 12 mg/kg), 15-acetyl-DON (0.5 mg/kg), and zearalenone (0.6 mg/kg) were identified in the contaminated diets arising from contaminated grains grown in Ontario, Canada. The concentrations of DON arising from naturally contaminated grains in this study were similar to purified mycotoxin fed to experimental mice. The chronic feeding of Fusarium mycotoxins induced small decreases in hematocrit values, total numbers of white blood cells, lymphocytes including both CD4+ and CD8+ T lymphocytes and B lymphocytes, and biliary IgA concentration. Supplementation of diets containing feedborne mycotoxins with GMA prevented the reduction in total number of B lymphocytes in the peripheral blood and the reduction in biliary IgA concentration. In addition, the delayed-type hypersensitivity response to dinitrochlorobenzene was increased by feed-borne mycotoxins, whereas IgG and IgM antibody titers to sheep red blood cells were not affected by diet. We concluded that chronic consumption of grains naturally contaminated with Fusarium mycotoxins at levels likely to be encountered in practice were not systemically immunosuppressive or hematotoxic; however, mucosal immunocompetence needs to be explored further.
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Dänicke, Sven; Pappritz, Julia; Goyarts, Tanja; Xu, Bu; Rautenschlein, Silke
2011-02-01
Two experiments were carried out to examine the effects of feeding an uncontaminated control diet (CON) or a Fusarium toxin-contaminated diet (FUS; 10.7 mg deoxynivalenol [DON]/kg diet) to growing broilers, which were either uninfected or infected with infectious bursal disease virus (IBDV) beginning at 1 day post hatch. Broilers had been infected at three weeks post hatch with either a classical virulent infectious bursal disease virus (IBDV-IM, Exp. 1) or a very virulent IBDV (vvIBDV, Exp. 2) strain. The effects of the DON-contaminated diet in combination with the virus-infection on the bursa of Fabricius and spleen were determined at 3 and 6-7 days post infection. The transient development of the bursa oedema and the bursa atrophy was not significantly affected by the diet after infection with the different IBDV-strains. The histopathological lesions were more severe in IBDV-IM-infected birds at 6 days post infection when additionally exposed to the FUS diet as compared to the FUS-free feed. Most parameters of the bursa of Fabricius and spleen protein turnover (e.g. fractional protein synthesis rate, protein, DNA and RNA content and derived indices) were significantly and interactively influenced by infection and stage of infection. The vvIBDV-infected birds responded with a more pronounced depressing effect on the fractional protein synthesis rate after feeding the DON-containing FUS diet when compared to their IBDV-IM-infected counterparts, where the opposite effect was observed. It can be concluded that feeding a FUS diet to IBDV-infected broilers might modulate the virulence-dependent pathogenesis of an IBDV infection.
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Burt, C; Steed, A; Gosman, N; Lemmens, M; Bird, N; Ramirez-Gonzalez, R; Holdgate, S; Nicholson, P
2015-09-01
Markers closely flanking a Type 1 FHB resistance have been produced and the potential of combining this with Type 2 resistances to improve control of FHB has been demonstrated. Two categories of resistance to Fusarium head blight (FHB) in wheat are generally recognised: resistance to initial infection (Type 1) and resistance to spread within the head (Type 2). While numerous sources of Type 2 resistance have been reported, relatively fewer Type 1 resistances have been characterised. Previous study identified a Type 1 FHB resistance (QFhs.jic-4AS) on chromosome 4A in Triticum macha. Little is known about the effect of combining Type 1 and Type 2 resistances on overall FHB symptoms or accumulation of the mycotoxin deoxynivalenol (DON). QFhs.jic-4AS was combined independently with two Type 2 FHB resistances (Fhb1 and one associated with the 1BL/1RS translocation). While combining Type 1 and Type 2 resistances generally reduced visual symptom development, the effect on DON accumulation was marginal. A lack of polymorphic markers and a limited number of recombinants had originally prevented accurate mapping of the QFhs.jic-4AS resistance. Using an array of recently produced markers in combination with new populations, the position of QFhs.jic-4AS has been determined to allow this resistance to be followed in breeding programmes.
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Ajandouz, El Hassan; Berdah, Stéphane; Moutardier, Vincent; Bege, Thierry; Birnbaum, David Jérémie; Perrier, Josette; Di Pasquale, Eric; Maresca, Marc
2016-01-01
In addition to deoxynivalenol (DON), acetylated derivatives, i.e., 3-acetyl and 15-acetyldexynivalenol (or 3/15ADON), are present in cereals leading to exposure to these mycotoxins. Animal and human studies suggest that 3/15ADON are converted into DON after their ingestion through hydrolysis of the acetyl moiety, the site(s) of such deacetylation being still uncharacterized. We used in vitro and ex vivo approaches to study the deacetylation of 3/15ADON by enzymes and cells/tissues present on their way from the food matrix to the blood in humans. We found that luminal deacetylation by digestive enzymes and bacteria is limited. Using human cells, tissues and S9 fractions, we were able to demonstrate that small intestine and liver possess strong deacetylation capacity compared to colon and kidneys. Interestingly, in most cases, deacetylation was more efficient for 3ADON than 15ADON. Although we initially thought that carboxylesterases (CES) could be responsible for the deacetylation of 3/15ADON, the use of pure human CES1/2 and of CES inhibitor demonstrated that CES are not involved. Taken together, our original model system allowed us to identify the small intestine and the liver as the main site of deacetylation of ingested 3/15ADON in humans. PMID:27483321
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The AtNFXL1 gene functions as a signaling component of the type A trichothecene-dependent response
Asano, Tomoya; Yasuda, Michiko; Nakashita, Hideo; Kimura, Makoto; Yamaguchi1, Kazuo
2008-01-01
Phytopathogenic Fusarium species produce the trichothecene family of phytotoxins, which function as a virulence factor during infection of plants. Trichothecenes are classifiable into four major groups by their chemical structures. Recently, the AtNFXL1 gene was reported as a type A trichothecene T-2 toxin-inducible gene. The AtNFXL1 gene encodes a putative transcription factor with similarity to the human transcription repressor NF-X1. The atnfxl1 mutant exhibited hypersensitivity phenotype to T-2 toxin but not to type B deoxynivalenol (DON) in comparison with wild type when Arabidopsis thaliana grew on agar medium containing trichothecenes. The absence or presence of a carbonyl group at the C8 position distinguishes type A and type B. Growth defect by another type A trichothecene diacetoxyscirpenol (DAS), was weakly enhanced in the atnfxl1 mutant. Diacetoxyscirpenol is distinguishable from T-2 toxin only by the absence of an isovaleryl group at the C8 position. Correspondingly, the AtNFXL1 promoter activity was apparently induced in T-2 toxin-treated and DAS-treated plants. In contrast, DON failed to induce the AtNFXL1 promoter activity. Consequently, the AtNFXL1 gene functions as a signaling component of the type A trichothecene-dependent response in Arabidopsis. In addition, the C8 position of trichothecenes might be closely related to the function of AtNFXL1 gene. PMID:19704430
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Oocyte quality in mice is affected by a mycotoxin-contaminated diet.
Hou, Yan-Jun; Xiong, Bo; Zheng, Wei-Jiang; Duan, Xing; Cui, Xiang-Shun; Kim, Nam-Hyung; Wang, Qiang; Xu, Yin-Xue; Sun, Shao-Chen
2014-05-01
Mycotoxins, such as deoxynivalenol (DON), zearalenone (ZEN), and aflatoxin (AF), are commonly found in many food commodities and may impair the growth and reproductive efficiency of animals and humans. We investigated the effects of a mycotoxin-contaminated diet on mouse oocyte quality. Maize contaminated with DON (3.875 mg/kg), ZEN (1,897 μg/kg), and AF (806 μg/kg) was incorporated into a mouse diet at three different levels (0, 15, and 30% w/w). After 4 weeks, ovarian and germinal vesicle oocyte indices decreased in mycotoxin-fed mice. Oocytes from these mice exhibited low developmental competence with reduced germinal vesicle breakdown and polar body extrusion rates. Embryo developmental competence also showed a similar pattern, and the majority of embryos could not develop to the morula stage. Actin expression was also reduced in both the oocyte cortex and cytoplasm, which was accompanied by decreased expression of the actin nucleation factors profilin-1 and mDia1. Moreover, a large percentage of oocytes derived from mice that were fed a mycotoxin-contaminated diet exhibited aberrant spindle morphology, a loss of the cortical granule-free domain, and abnormal mitochondrial distributions, which further supported the decreased oocyte quality. Thus, our results demonstrate that mycotoxins are toxic to the mouse reproductive system by affecting oocyte quality. Copyright © 2013 Wiley Periodicals, Inc.
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Krysińska-Traczyk, Ewa; Perkowski, Juliusz; Kostecki, Marian; Dutkiewicz, Jacek; Kiecana, Irena
2003-01-01
The studies to determine the level of filamentous fungi and mycotoxins were carried out in samples of grain and grain dust during threshing of cereals by a combine harvester. High concentration of fungi was noted in grain and grain dust samples, it ranged from 5.0 to 520.0 cfu/g.10(3) and from 275.0 to 2825.0 cfu/g.10(3), respectively Allergizing and toxigenic fungi of Alternaria, Geotrichum, Cladosporium, Penicillium, Aspergillus and Fusarium species were observed in the study samples of grain and grain dust. In the samples of wheat grain, mycotoxins were also noted: moniliformin (MON), deoxynivalenol (DON) and ochratoxin A (OTA); their concentrations ranged from 0.025 to 0.088 microgram/g; 0.015-0.068 microgram/g; and from 0.0004 to 0.0008 microgram/g, respectively. The level of mycotoxins in the grain dust samples was within the range of 0.025-0.149 microgram/g-MON; 0.015-0.215 microgram/g-DON; 0.015-0.360 microgram/g-NIV; and 0.0004-0.0012 microgram/g-OTA. A significant correlation was observed between the occurrence of fungi of Fusarium species and the concentration of pathologic mycotoxins. The results confirm a considerable occupational risk among farmers engaged in grain threshing due to inhalation of pathogenic species of filamentous fungi and mycotoxins.
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Mycotoxins – Limits and Regulations
Mazumder, Papiya Mitra; Sasmal, D.
2001-01-01
Since early years, a need has always been felt for some control on the quality of foodstuffs. With the discovery of aflatoxins in the early sixties, health authorities in man countries have become active in establishing regulations to protect their citizens and livestock fro t potential harm caused by mycotoxins. FDA mycotox-ins-in-foods sampling program is continuing with an objective to remove those foods from interstate commerce that contain Aflatoxins “at levels judged to be of regulator significance” Aflatoxins, Fumonisin B1 and B2, Deoxynivalenol (DON) Ochratoxin A and Patulin occur in a number of food products. FDA workers were instructed to sample and analyze all products for different types of mycotoxins. All baby foods should always be analyzed for all type of mycotoxins. The limits of Aflatoxins B1,B2,! < G2, and M1 in foods and feed stuffs varies from (0-40) ppb for foods & 0-1000ppb for food); for Ochratoxin A(0-50 ppb in food and 0-1000ppb in feed); for Don (500-2000ppb in food & 5-10,000 ppb in feed); for Zearalenone (0-1000 ppb in food); for Patulin (0-50 ppb in foods), for Diacetoxyscirpenol (0-100 ppd in feed); for chetomin (0ppb I feed); for stachybotryotoxin (0ppb in feeds and for Fumonisins (0-1000 ppb in food 5000-50,000 ppb in feedstuffs). PMID:22557007
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7 CFR 868.90 - Fees for certain Federal inspection services.
Code of Federal Regulations, 2014 CFR
2014-01-01
... Laboratory Test Services 1 Laboratory tests Fees (1) Aflatoxin (Quantitative—HPLC) $182.00 (2) Aflatoxin (Quantitative—Test Kit) 87.00 (3) Aflatoxin (Qualitative—Test Kit) 47.00 (4) Appearance and odor 7.00 (5) Ash 17...) Vomitoxin (Quantitative—Test Kit) 81.00 (32) Other laboratory analytical services (per hour per service...
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7 CFR 868.90 - Fees for certain Federal inspection services.
Code of Federal Regulations, 2011 CFR
2011-01-01
... Laboratory Test Services 1 Laboratory tests Fees (1) Aflatoxin (Quantitative—HPLC) $182.00 (2) Aflatoxin (Quantitative—Test Kit) 87.00 (3) Aflatoxin (Qualitative—Test Kit) 47.00 (4) Appearance and odor 7.00 (5) Ash 17...) Vomitoxin (Quantitative—Test Kit) 81.00 (32) Other laboratory analytical services (per hour per service...
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7 CFR 868.90 - Fees for certain Federal inspection services.
Code of Federal Regulations, 2012 CFR
2012-01-01
... Laboratory Test Services 1 Laboratory tests Fees (1) Aflatoxin (Quantitative—HPLC) $182.00 (2) Aflatoxin (Quantitative—Test Kit) 87.00 (3) Aflatoxin (Qualitative—Test Kit) 47.00 (4) Appearance and odor 7.00 (5) Ash 17...) Vomitoxin (Quantitative—Test Kit) 81.00 (32) Other laboratory analytical services (per hour per service...
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7 CFR 868.90 - Fees for certain Federal inspection services.
Code of Federal Regulations, 2013 CFR
2013-01-01
... Laboratory Test Services 1 Laboratory tests Fees (1) Aflatoxin (Quantitative—HPLC) $182.00 (2) Aflatoxin (Quantitative—Test Kit) 87.00 (3) Aflatoxin (Qualitative—Test Kit) 47.00 (4) Appearance and odor 7.00 (5) Ash 17...) Vomitoxin (Quantitative—Test Kit) 81.00 (32) Other laboratory analytical services (per hour per service...
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Yoshizawa, Takumi; Morooka, Nobuichi
1975-01-01
Attempts were made to elucidate the acetyl transformation of novel trichothecene mycotoxins, 3α,7α,15-trihydroxy-12,13-epoxytrichothec-9-en-8-one (deoxynivalenol) and its derivatives, by trichothecene-producing strains of Fusarium nivale, F. roseum, and F. solani. In the peptone-supplemented Czapek-Dox medium, F. roseum converted 3α-acetoxy-7α,15-dihydroxy-12,13-epoxytrichothec-9-en-8-one (3-acetyldeoxynivalenol) to deoxynivalenol. 3-Acetyldeoxynivalenol was also deacetylated by intact mycelia of the three strains in sugar-free Czapek-Dox medium. The growing F. nivale acetylated deoxynivalenol to afford a small amount of 3-acetyldeoxynivalenol. 3α,7α,15-Triacetoxy-12,13-epoxytrichothec-9-en-8-one (deoxynivalenol triacetate) was transformed by the intact mycelium of F. solani into 7α,15-diacetoxy-3α-hydroxy-12,13-epoxytrichothec-9-en-8-one (7,15-diacetyl-deoxynivalenol), which was then deacetylated to give 7α-acetoxy-3α,15-dihydroxy-12,13-epoxytrichothec-9-en-8-one (7-acetyldeoxynivalenol). It was noted that the ester at C-7 was not hydrolyzed by the fungal mycelium. PMID:234156
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Fate of enniatins and deoxynivalenol during pasta cooking.
de Nijs, Monique; van den Top, Hester; de Stoppelaar, Joyce; Lopez, Patricia; Mol, Hans
2016-12-15
The fate of deoxynivalenol and enniatins was studied during cooking of commercially available dry pasta in the Netherlands in 2014. Five samples containing relatively high levels of deoxynivalenol and/or enniatins were selected for the cooking experiment. Cooking was performed in duplicate on different days, under standardised conditions, simulating house-hold preparation. Samples were extracted with a mixture of acetonitrile/water followed by salt-induced partitioning. The extracts were analysed by LC-MS/MS. The method limits of detection were 8μg/kg for deoxynivalenol, 10μg/kg for enniatin A1 and 5μg/kg for enniatins A, B and B1. During the cooking of the five dry pasta samples, 60% of the deoxynivalenol and 83-100% of the enniatins were retained in the cooked pasta. It is recommended to study food processing fate of mycotoxins through naturally contaminated materials (incurred materials). Copyright © 2016 Elsevier Ltd. All rights reserved.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Del Regno, Marisanta; Adesso, Simona; Popolo, Ada
Mycotoxins are secondary fungal metabolites often found as contaminants in almost all agricultural commodities worldwide, and the consumption of food or feed contaminated by mycotoxins represents a major risk for human and animal health. Reactive oxygen species are normal products of cellular metabolism. However, disproportionate generation of reactive oxygen species poses a serious problem to bodily homeostasis and causes oxidative tissue damage. In this study we analyzed the effect of two trichothecenes mycotoxins: nivalenol and deoxynivalenol, alone and in combination, on oxidative stress in the non-tumorigenic intestinal epithelial cell line IEC-6. Our results indicate the pro-oxidant nivalenol effect in IEC-6,more » the stronger pro-oxidant effect of nivalenol when compared to deoxynivalenol and, interestingly, that nivalenol increases deoxynivalenol pro-oxidative effects. Mechanistic studies indicate that the observed effects were mediated by NADPH oxidase, calcium homeostasis alteration, NF-kB and Nrf2 pathways activation and by iNOS and nitrotyrosine formation. The toxicological interaction by nivalenol and deoxynivalenol reported in this study in IEC-6, points out the importance of the toxic effect of these mycotoxins, mostly in combination, further highlighting the risk assessment process of these toxins that are of growing concern. - Highlights: • Nivalenol induces oxidative stress in intestinal epithelial cells (IECs). • Nivalenol increases deoxynivalenol pro-oxidant effects in IECs. • Nivalenol and deoxynivalenol trigger antioxidant response IECs. • These results indicate the importance of mycotoxins co-contamination.« less
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Phoku, J Z; Barnard, T G; Potgieter, N; Dutton, M F
2017-04-01
A study on the potential of houseflies (Musca domestica L.) to spread fungal spores in Gauteng Province, South Africa proved that houseflies are vectors for fungal spores. Therefore, there is a need to determine the toxigenic potentials and to identify the mycotoxins produced by fungal isolates derived from this study. In total 377 potentially toxigenic isolates of Aspergillus (186), Fusarium (85) and Penicillium (106) species (spp.) were isolated. These isolates were further tested for their ability to produce aflatoxins (AFs) [aflatoxin B 1 , B 2 , G 1 and G 2 ], deoxynivalenol (DON), fumonisin B 1 (FB 1 ) ochratoxin A (OTA), and zearalenone (ZEA) by high-performance liquid chromatography (HPLC) respectively. Strains of A. flavus and A. parasiticus belonging to the genera of Aspergillus were found to be the main producers of AFB 1 , AFB 2 , AFG 1 , and AFG 2 , while A. carbonarius, A. niger and A. ochraceus produced OTA. Fumonisin B 1 was produced by F. verticillioides and F. proliferatum with concentrations ranging from 20 to 1834μg/kg and 79 to 262μg/kg respectively. Deoxynivalenol produced mainly by F. culmorum (2-6μg/kg), F. graminearum (1-4μg/kg), F. poae (1-3μg/kg), and F. sporotrichioides (2-3μg/kg) species was the least detected toxin in this study. The high mycotoxins levels produced in isolates from houseflies in this study are regarded as unsafe, especially when international legislated tolerance levels for mycotoxins are considered. Thus, possible human exposure to mycotoxins may pose concerns with respect to human health and demands constant and consistent investigation. Copyright © 2017 Elsevier B.V. All rights reserved.
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Occurrence of 26 Mycotoxins in the Grain of Cereals Cultivated in Poland
Bryła, Marcin; Waśkiewicz, Agnieszka; Podolska, Grażyna; Szymczyk, Krystyna; Jędrzejczak, Renata; Damaziak, Krzysztof; Sułek, Alicja
2016-01-01
The levels of 26 mycotoxins were determined in 147 samples of the grain of cereals cultivated in five regions of Poland during the 2014 growing season. The HPLC-HRMS (time-of-flight) analytical technique was used. An analytical procedure to simultaneously determine 26 mycotoxins in grain was developed, tested and verified. Samples from eastern and southern Poland were more contaminated with mycotoxins than the samples from northern and western Poland. Toxins produced by Fusarium fungi were the main contaminants found. Some deoxynivalenol (DON) was found in 100% of the tested samples of wheat (Osiny, Borusowa, Werbkowice), triticale, winter barley and oats, while the maximum permissible DON level (as defined in the EU Commission Regulation No. 1881/2006) was exceeded in 10 samples. Zearalenone (ZEN), DON metabolites and enniatins were also commonly found. The presence of mycotoxins in grain reflected the prevailing weather conditions during the plant flowering/earing stages, which were favorable for the development of blight. Among all investigated wheat genotypes, cv. Fidelius was the least contaminated, while Bamberka, Forkida and Kampana were the most contaminated. However, the single-factor ANOVA analysis of variance did not reveal (at a statistical significance level α = 0.05) any differences between levels of mycotoxins in individual genotypes. Triticale was the most contaminated grain among all of the tested varieties. ZEN, DON and the sum of 3-acetyldexynivalenol and 15-acetyldeoxynivalenol (3- and 15-ADON) were found in 100% of the tested triticale samples at concentrations within the 4–86, 196–1326 and 36–374 µg·kg−1 range, respectively. Of particular concern was the fact that some “emerging mycotoxins” (enniatins) (in addition to commonly-known and legally-regulated mycotoxins) were also found in the tested triticale samples (enniatin B (Enn-B), enniatin B1 (Enn-B1), enniatin A-1 (Enn-A1), 100% of samples, and enniatin A (Enn-A), 70% of samples). Depending on the toxin, they were found at levels between 8 and 3328 µg·kg−1. PMID:27231939
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Rodríguez-Carrasco, Yelko; Fattore, Margherita; Albrizio, Stefania; Berrada, Houda; Mañes, Jordi
2015-07-01
Since cereals are raw materials for production of beer and beer-based drinks, the occurrence mycotoxins in 154 beer samples was topic of investigation in this study. The analyses were conducted using QuEChERS extraction and gas chromatography-tandem mass spectrometry determination. The analytical method showed recoveries for vast majority of analytes ranged from 70% to 110%, relative standard deviations lower than 15% and limits of detection from 0.05 to 8 μg/L. A significant incidence of HT-2 toxin and deoxynivalenol (DON) were found in 9.1% and 59.7% of total samples, respectively. The exposure of European population to mycotoxins through beer consumption was assessed. No toxicological concern was associated to mycotoxins exposure for average beer consumers. Despite that, for heavy beer drinkers, the contribution of this commodity to the daily intake is not negligible, approaching or even exceeding the safety levels. Copyright © 2015 Elsevier Ltd. All rights reserved.
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Multi-Toxic Endpoints of the Foodborne Mycotoxins in Nematode Caenorhabditis elegans
Yang, Zhendong; Xue, Kathy S.; Sun, Xiulan; Tang, Lili; Wang, Jia-Sheng
2015-01-01
Aflatoxins B1 (AFB1), deoxynivalenol (DON), fumonisin B1 (FB1), T-2 toxin (T-2), and zearalenone (ZEA) are the major foodborne mycotoxins of public health concerns. In the present study, the multiple toxic endpoints of these naturally-occurring mycotoxins were evaluated in Caenorhabditis elegans model for their lethality, toxic effects on growth and reproduction, as well as influence on lifespan. We found that the lethality endpoint was more sensitive for T-2 toxicity with the EC50 at 1.38 mg/L, the growth endpoint was relatively sensitive for AFB1 toxic effects, and the reproduction endpoint was more sensitive for toxicities of AFB1, FB1, and ZEA. Moreover, the lifespan endpoint was sensitive to toxic effects of all five tested mycotoxins. Data obtained from this study may serve as an important contribution to knowledge on assessment of mycotoxin toxic effects, especially for assessing developmental and reproductive toxic effects, using the C. elegans model. PMID:26633509
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An overview of the toxicology and toxicokinetics of fusarenon-X, a type B trichothecene mycotoxin.
Aupanun, Sawinee; Poapolathep, Saranya; Giorgi, Mario; Imsilp, Kanjana; Poapolathep, Amnart
2017-01-20
Fusarenon-X (FX) is a type B trichothecene mycotoxin that is frequently observed along with deoxynivalenol (DON) and nivalenol (NIV) in agricultural commodities. This review aims to give an overview of the literature concerning the toxicology and toxicokinetics of FX. FX is primarily found in cereals grown in temperate regions, but it can also be found worldwide because of the global transport of products. The major toxicity of FX occurs through inhibition of protein synthesis, followed by the disruption of DNA synthesis. Moreover, FX has also been shown to induce apoptosis in in vitro and in vivo studies. The targets of FX are organs containing actively proliferating cells, such as the thymus, spleen, skin, small intestine, testes and bone marrow. FX causes immunosuppression, intestinal malabsorption, developmental toxicity and genotoxicity. In addition, sufficient evidence of carcinogenicity in experimental animals is currently lacking, and the International Agency for Research on Cancer (IARC) classifies it as a group 3 carcinogen.
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Dhariwal, Raman; Fedak, George; Dion, Yves; Pozniak, Curtis; Laroche, André; Eudes, François; Randhawa, Harpinder Singh
2018-01-01
Triticale (xTriticosecale Wittmack) is an important feed crop which suffers severe yield, grade and end-use quality losses due to Fusarium head blight (FHB). Development of resistant triticale cultivars is hindered by lack of effective genetic resistance sources. To dissect FHB resistance, a doubled haploid spring triticale population produced from the cross TMP16315/AC Ultima using a microspore culture method, was phenotyped for FHB incidence, severity, visual rating index (VRI), deoxynivalenol (DON) and some associated traits (ergot, grain protein content, test weight, yield, plant height and lodging) followed by single nucleotide polymorphism (SNP) genotyping. A high-density map consisting of 5274 SNPs, mapped on all 21 chromosomes with a map density of 0.48 cM/SNP, was constructed. Together, 17 major quantitative trait loci were identified for FHB on chromosomes 1A, 2B, 3A, 4A, 4R, 5A, 5R and 6B; two of incidence loci (on 2B and 5R) also co-located with loci for severity and VRI, and two other loci of VRI (on 1A and 4R) with DON accumulation. Major and minor loci were also identified for all other traits in addition to many epistasis loci. This study provides new insight into the genetic basis of FHB resistance and their association with other traits in triticale. PMID:29304028
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Fusarium and mycotoxin spectra in Swiss barley are affected by various cropping techniques.
Schöneberg, Torsten; Martin, Charlotte; Wettstein, Felix E; Bucheli, Thomas D; Mascher, Fabio; Bertossa, Mario; Musa, Tomke; Keller, Beat; Vogelgsang, Susanne
2016-10-01
Fusarium head blight is one of the most important cereal diseases worldwide. Cereals differ in terms of the main occurring Fusarium species and the infection is influenced by various factors, such as weather and cropping measures. Little is known about Fusarium species in barley in Switzerland, hence harvest samples from growers were collected in 2013 and 2014, along with information on respective cropping factors. The incidence of different Fusarium species was obtained by using a seed health test and mycotoxins were quantified by LC-MS/MS. With these techniques, the most dominant species, F. graminearum, and the most prominent mycotoxin, deoxynivalenol (DON), were identified. Between the three main Swiss cropping systems, Organic, Extenso and Proof of ecological performance, we observed differences with the lowest incidence and toxin accumulation in organically cultivated barley. Hence, we hypothesise that this finding was based on an array of growing techniques within a given cropping system. We observed that barley samples from fields with maize as previous crop had a substantially higher F. graminearum incidence and elevated DON accumulation compared with other previous crops. Furthermore, the use of reduced tillage led to a higher disease incidence and toxin content compared with samples from ploughed fields. Further factors increasing Fusarium infection were high nitrogen fertilisation as well as the application of fungicides and growth regulators. Results from the current study can be used to develop optimised cropping systems that reduce the risks of mycotoxin contamination.
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Czembor, Elżbieta; Stępień, Łukasz; Waśkiewicz, Agnieszka
2015-01-01
Maize is one of the most important crops and Poland is the fifth largest producing country in Europe. Diseases caused by Fusarium spp. can affect the yield and grain quality of maize because of contamination with numerous mycotoxins produced by these fungi. The present study was performed to identify the prevailing Fusarium species and the environmental factors affecting their frequencies and the contamination of grain with the main mycotoxins deoxynivalenol (DON), zearalenone (ZON) and fumonisin B1 (FB1). Thirty kernel samples were collected in three locations in 2011 and in seven locations in 2012 from three hybrids. On average, 25.24% kernels were colonized by Fusarium spp. (424 strains were isolated). Fusarium verticillioides and F. temperatum were the most prevalent species, F. subglutinans, F. proliferatum and F. graminearum were in minor abundance. In total, 272 isolates of F. verticillioides and 81 isolates of F. temperatum were identified. Fusarium temperatum frequency ranged from 1.70% to 28.57% and differences between locations were significant. Fumonisin B1 was found in all tested samples. DON was found in 66.67% and ZON in 43.33% of samples. Rainfall amount positively affected F. temperatum and F. subglutinans frequency in opposite to mean temperatures in July. On the other hand, relationships between frequency of these species and historical data from 1950–2000 for annual temperature range were negative in contrast to the coldest quarter temperatures. PMID:26225823
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Fusarium and mycotoxin spectra in Swiss barley are affected by various cropping techniques
Schöneberg, Torsten; Martin, Charlotte; Wettstein, Felix E.; Bucheli, Thomas D.; Mascher, Fabio; Bertossa, Mario; Musa, Tomke; Keller, Beat; Vogelgsang, Susanne
2016-01-01
ABSTRACT Fusarium head blight is one of the most important cereal diseases worldwide. Cereals differ in terms of the main occurring Fusarium species and the infection is influenced by various factors, such as weather and cropping measures. Little is known about Fusarium species in barley in Switzerland, hence harvest samples from growers were collected in 2013 and 2014, along with information on respective cropping factors. The incidence of different Fusarium species was obtained by using a seed health test and mycotoxins were quantified by LC-MS/MS. With these techniques, the most dominant species, F. graminearum, and the most prominent mycotoxin, deoxynivalenol (DON), were identified. Between the three main Swiss cropping systems, Organic, Extenso and Proof of ecological performance, we observed differences with the lowest incidence and toxin accumulation in organically cultivated barley. Hence, we hypothesise that this finding was based on an array of growing techniques within a given cropping system. We observed that barley samples from fields with maize as previous crop had a substantially higher F. graminearum incidence and elevated DON accumulation compared with other previous crops. Furthermore, the use of reduced tillage led to a higher disease incidence and toxin content compared with samples from ploughed fields. Further factors increasing Fusarium infection were high nitrogen fertilisation as well as the application of fungicides and growth regulators. Results from the current study can be used to develop optimised cropping systems that reduce the risks of mycotoxin contamination. PMID:27491813
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Tamura, Masayoshi; Mochizuki, Naoki; Nagatomi, Yasushi; Harayama, Koichi; Toriba, Akira; Hayakawa, Kazuichi
2015-01-01
A high-resolution liquid chromatography-Orbitrap mass spectrometry (LC-Orbitrap MS) method was developed for simultaneous determination of 20 Fusarium toxins (nivalenol, fusarenon-X, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, HT-2 toxin, T-2 toxin, neosolaniol, diacetoxyscirpenol, fumonisin B1, fumonisin B2, fumonisin B3, fumonisin A1, fumonisin A2, fumonisin A3, zearalenone, α-zearalenol, β-zearalenol, α-zearalanol, and β-zearalanol) in cereals. The separation of 20 Fusarium toxins with good peak shapes was achieved using a pentafluorophenyl column, and Orbitrap MS was able to detect accurately from cereal matrix components within ±0.77 ppm. The samples were prepared using a QuEChERS kit for extraction and a multifunctional cartridge for purification. The linearity, repeatability, and recovery of the method were >0.9964, 0.8%–14.7%, and 71%–106%, respectively. Using this method, an analysis of 34 commercially available cereals detected the presence of deoxynivalenol, 15-acetyl deoxynivalenol, fumonisin B1, fumonisin B2, fumonisin B3, fumonisn A1, fumonisin A2, fumonisin A3, and zearalenone in corn samples with high concentration and frequency. Trichothecenes was detected from wheat samples with high frequency; in particular, the concentration of deoxynivalenol was high. Conversely, α-zearalenol, β-zearalenol, α-zearalanol, and β-zearalanol were not detected in any of the samples. PMID:26008230
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Multi-mycotoxin stable isotope dilution LC-MS/MS method for Fusarium toxins in beer.
Habler, Katharina; Gotthardt, Marina; Schüler, Jan; Rychlik, Michael
2017-03-01
A stable isotope dilution LC-MS/MS multi-mycotoxin method was developed for 12 different Fusarium toxins including modified mycotoxins in beer (deoxynivalenol-3-glucoside, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyl-deoxynivalenol, HT2-toxin, T2-toxin, enniatin B, B1, A1, A, beauvericin and zearalenone). As sample preparation and purification of beer a combined solid phase extraction for trichothecenes, enniatins, beauvericin and zearalenone was firstly developed. The validation of the new method gave satisfying results: intra-day and inter-day precision and recoveries were 1-5%, 2-8% and 72-117%, respectively. In total, 61 different organic and conventional beer samples from Germany and all over the world were analyzed by using the newly developed multi-mycotoxin method. In summary, deoxynivalenol, deoxynivalenol-3-glucoside, 3-acetyldeoxynivaleneol and enniatin B were quantified in rather low contents in the investigated beer samples. None of the other monitored Fusarium toxins like 15-acetyldeoxynivalenol, HT2- and T2-toxin, zearalenone, enniatin B1, A1, A or beauvericin were detectable. Copyright © 2016 Elsevier Ltd. All rights reserved.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Alassane-Kpembi, Imourana; Université de Toulouse, ENVT, INP, UMR 1331 Toxalim, F-31076 Toulouse; Institut des Sciences Biomédicales Appliquées, Cotonou, Bénin
Deoxynivalenol (DON) is the most prevalent trichothecene mycotoxin in crops in Europe and North America. DON is often present with other type B trichothecenes such as 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV) and fusarenon-X (FX). Although the cytotoxicity of individual mycotoxins has been widely studied, data on the toxicity of mycotoxin mixtures are limited. The aim of this study was to assess interactions caused by co-exposure to Type B trichothecenes on intestinal epithelial cells. Proliferating Caco-2 cells were exposed to increasing doses of Type B trichothecenes, alone or in binary or ternary mixtures. The MTT test and neutral red uptake,more » respectively linked to mitochondrial and lysosomal functions, were used to measure intestinal epithelial cytotoxicity. The five tested mycotoxins had a dose-dependent effect on proliferating enterocytes and could be classified in increasing order of toxicity: 3-ADON < 15-ADON ≈ DON < NIV ≪ FX. Binary or ternary mixtures also showed a dose-dependent effect. At low concentrations (cytotoxic effect between 10 and 30–40%), mycotoxin combinations were synergistic; however DON–NIV–FX mixture showed antagonism. At higher concentrations (cytotoxic effect around 50%), the combinations had an additive or nearly additive effect. These results indicate that the simultaneous presence of low doses of mycotoxins in food commodities and diet may be more toxic than predicted from the mycotoxins alone. Considering the frequent co-occurrence of trichothecenes in the diet and the concentrations of toxins to which consumers are exposed, this synergy should be taken into account. - Highlights: • We assessed the individual and combined cytotoxicity of five trichothecenes. • The tested concentrations correspond to the French consumer exposure levels. • The type of interaction in combined cytotoxicity varied with the effect level. • Low doses of Type B trichothecenes induced synergistic cytotoxicity. • Ternary combination DON–NIV–FX showed antagonism.« less
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Carere, Jason; Fitzgerald, Timothy L.; Stiller, Jiri; Covarelli, Lorenzo; Xu, Qian; Gubler, Frank; Colgrave, Michelle L.; Gardiner, Donald M.; Manners, John M.; Henry, Robert J.; Kazan, Kemal
2017-01-01
Abstract Background and Aims Fusarium crown rot caused by the fungal pathogen Fusarium pseudograminearum is a disease of wheat and barley, bearing significant economic cost. Efforts to develop effective resistance to this disease have been hampered by the quantitative nature of resistance and a lack of understanding of the factors associated with resistance and susceptibility. Here, we aimed to dissect transcriptional responses triggered in wheat by F. pseudograminearum infection. Methods We used an RNA-seq approach to analyse host responses during a compatible interaction and identified >2700 wheat genes differentially regulated after inoculation with F. pseudograminearum. The production of a few key metabolites and plant hormones in the host during the interaction was also analysed. Key Results Analysis of gene ontology enrichment showed that a disproportionate number of genes involved in primary and secondary metabolism, signalling and transport were differentially expressed in infected seedlings. A number of genes encoding pathogen-responsive uridine-diphosphate glycosyltransferases (UGTs) potentially involved in detoxification of the Fusarium mycotoxin deoxynivalenol (DON) were differentially expressed. Using a F. pseudograminearum DON-non-producing mutant, DON was shown to play an important role in virulence during Fusarium crown rot. An over-representation of genes involved in the phenylalanine, tryptophan and tyrosine biosynthesis pathways was observed. This was confirmed through metabolite analyses that demonstrated tryptamine and serotonin levels are induced after F. pseudograminearum inoculation. Conclusions Overall, the observed host response in bread wheat to F. pseudograminearum during early infection exhibited enrichment of processes related to pathogen perception, defence signalling, transport and metabolism and deployment of chemical and enzymatic defences. Additional functional analyses of candidate genes should reveal their roles in disease resistance or susceptibility. Better understanding of host responses contributing to resistance and/or susceptibility will aid the development of future disease improvement strategies against this important plant pathogen. PMID:27941094
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Powell, Jonathan J; Carere, Jason; Fitzgerald, Timothy L; Stiller, Jiri; Covarelli, Lorenzo; Xu, Qian; Gubler, Frank; Colgrave, Michelle L; Gardiner, Donald M; Manners, John M; Henry, Robert J; Kazan, Kemal
2017-03-01
Fusarium crown rot caused by the fungal pathogen Fusarium pseudograminearum is a disease of wheat and barley, bearing significant economic cost. Efforts to develop effective resistance to this disease have been hampered by the quantitative nature of resistance and a lack of understanding of the factors associated with resistance and susceptibility. Here, we aimed to dissect transcriptional responses triggered in wheat by F. pseudograminearum infection. We used an RNA-seq approach to analyse host responses during a compatible interaction and identified >2700 wheat genes differentially regulated after inoculation with F. pseudograminearum . The production of a few key metabolites and plant hormones in the host during the interaction was also analysed. Analysis of gene ontology enrichment showed that a disproportionate number of genes involved in primary and secondary metabolism, signalling and transport were differentially expressed in infected seedlings. A number of genes encoding pathogen-responsive uridine-diphosphate glycosyltransferases (UGTs) potentially involved in detoxification of the Fusarium mycotoxin deoxynivalenol (DON) were differentially expressed. Using a F. pseudograminearum DON-non-producing mutant, DON was shown to play an important role in virulence during Fusarium crown rot. An over-representation of genes involved in the phenylalanine, tryptophan and tyrosine biosynthesis pathways was observed. This was confirmed through metabolite analyses that demonstrated tryptamine and serotonin levels are induced after F. pseudograminearum inoculation. Overall, the observed host response in bread wheat to F. pseudograminearum during early infection exhibited enrichment of processes related to pathogen perception, defence signalling, transport and metabolism and deployment of chemical and enzymatic defences. Additional functional analyses of candidate genes should reveal their roles in disease resistance or susceptibility. Better understanding of host responses contributing to resistance and/or susceptibility will aid the development of future disease improvement strategies against this important plant pathogen. © The Author 2016. Published by Oxford University Press on behalf of the Annals of Botany Company.
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Are Treated Celiac Patients at Risk for Mycotoxins? An Italian Case-Study
Cirlini, Martina; Mazzeo, Teresa; Roncoroni, Leda; Lombardo, Vincenza; Elli, Luca; Bardella, Maria T.; Agostoni, Carlo; Doneda, Luisa; Brighenti, Furio; Dall’Asta, Chiara; Pellegrini, Nicoletta
2016-01-01
Urinary biomarkers of mycotoxin exposure were evaluated in a group of celiac patients (n = 55) and in a control group of healthy subjects (n = 50) following their habitual diet. Deoxynivalenol (DON), zearalenone (ZEN), and fumonisin B1 (FB1) were monitored in 105 urinary samples collected from the two groups. Dietary habits were also recorded through compilation of a seven-day weighed dietary diary. Biomarkers of mycotoxin exposure were detected in 21 celiac patients and in 15 control subjects, corresponding to about 34% of total participants. In particular, ZEN was the most detected mycotoxin among all the studied subjects with a total of 19 positive cases. Results did not show a statistically significant difference in mycotoxin exposure between the two groups, and the presence of specific mycotoxins was not related to the intake of any particular food category. Our findings suggest little urgency of specific regulation for gluten free products, although the prevalence of exposure observed in free-living diets of both celiac and healthy subjects underlines the need of a constant surveillance on mycotoxins occurrence at large. PMID:28036017
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Krysińska-Traczyk, Ewa; Perkowski, Juliusz; Dutkiewicz, Jacek
2007-01-01
During combine harvesting of 5 various cereal crops (rye, barley, oats, buckwheat, corn) 24 samples of grain and 24 samples of settled grain dust were collected on farms located in the Lublin province of eastern Poland. The samples were examined for the concentration of total microfungi, Fusarium species, deoxynivalenol (DON), nivalenol (NIV), and ochratoxin A (OTA). Microfungi able to grow on malt agar were present in 79.2% of grain samples and in 91.7% of grain dust samples in the concentrations of 1.0-801.3x10(3) cfu/g and 1.5-12440.0x10(3) cfu/g, respectively. The concentration of microfungi in grain dust samples was significantly greater than in grain samples (p<0.01). Fusarium strains were isolated from 54.2% of grain samples and from 58.3% of grain dust samples in the concentrations of 0.1-375.0x10(3) cfu/g and 4.0-7,700.0x10(3) cfu/g, respectively. They were found in all samples of grain and grain dust from rye, barley and corn, but only in 0-16.7% of samples of grain and grain dust from oats and buckwheat. DON was found in 79.2% of grain samples and in 100% of grain dust samples in the concentrations of 0.001-0.18 microg/g and 0.006-0.283 microg/g, respectively. NIV was detected in 62.5% of grain samples and in 94.4% of grain dust samples in the concentrations of 0.004-0.502 microg/g and 0.005-0.339 microg/g, respectively. OTA was detected in 58.3% of grain samples and in 91.7% of grain dust samples in the concentrations of 0.00039- 0.00195 microg/g and 0.00036-0.00285 microg/g, respectively. The concentrations of DON, total fusariotoxins (DON+NIV) and OTA were significantly greater in grain dust samples than in grain samples (p<0.05, p<0.05, and p<0.001, respectively). The concentration of Fusarium poae in the samples of rye grain and dust was significantly correlated with the concentrations of DON (p<0.05), NIV (p<0.01), and total fusariotoxins (p<0.05). Similarly, the concentration of Fusarium culmorum in the samples of barley grain and dust was significantly correlated with the concentration of total fusariotoxins (p<0.05). A significant correlation was also found between the concentration of total fungi grown on malt agar and the concentration of OTA (p<0.05). In conclusion, although the concentration of DON, NIV and OTA in the samples of grain dust collected from 5 various cereals on farms in eastern Poland was not large, the persistent presence of these mycotoxins in over 90% of examined samples poses a potential health risk of chronic respiratory intoxication for exposed grain farmers.
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Microbial correlates of Fusarium biomass and deoxynivalenol content in individual wheat seeds
USDA-ARS?s Scientific Manuscript database
Manipulating the microbiome of wheat seeds and heads may contribute to control of Fusarium head blight and mycotoxin accumulation in grain, which creates a food safety hazard. With the aim of identifying novel management targets, we looked for correlations between Fusarium biomass or deoxynivalenol ...
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Puri, Krishna D; Yan, Changhui; Leng, Yueqiang; Zhong, Shaobin
2016-01-01
Fusarium graminearum is the major causal agent of Fusarium head blight (FHB) in barley and wheat in North America. The fungus not only causes yield loss of the crops but also produces harmful trichothecene mycotoxins [Deoxynivalenol (DON) and its derivatives-3-acetyldeoxynivalenol (3ADON) and 15-acetyldeoxynivalenol (15ADON), and nivalenol (NIV)] that contaminate grains. Previous studies showed a dramatic increase of 3ADON-producing isolates with higher aggressiveness and DON production than the 15ADON-producing isolates in North America. However, the genetic and molecular basis of differences between the two types of isolates is unclear. In this study, we compared transcriptomes of the 3ADON and 15ADON isolates in vitro (in culture media) and in planta (during infection on the susceptible wheat cultivar 'Briggs') using RNA-sequencing. The in vitro gene expression comparison identified 479 up-regulated and 801 down-regulated genes in the 3ADON isolates; the up-regulated genes were mainly involved in C-compound and carbohydrate metabolism (18.6%), polysaccharide metabolism (7.7%) or were of unknown functions (57.6%). The in planta gene expression analysis revealed that 185, 89, and 62 genes were up-regulated in the 3ADON population at 48, 96, and 144 hours after inoculation (HAI), respectively. The up-regulated genes were significantly enriched in functions for cellular import, C-compound and carbohydrate metabolism, allantoin and allantoate transport at 48 HAI, for detoxification and virulence at 96 HAI, and for metabolism of acetic acid derivatives, detoxification, and cellular import at 144 HAI. Comparative analyses of in planta versus in vitro gene expression further revealed 2,159, 1,981 and 2,095 genes up-regulated in the 3ADON isolates, and 2,415, 2,059 and 1,777 genes up-regulated in the 15ADON isolates at the three time points after inoculation. Collectively, our data provides a foundation for further understanding of molecular mechanisms involved in aggressiveness and DON production of the two chemotype isolates of F. graminearum.
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Dong, Fei; Qiu, Jianbo; Xu, Jianhong; Yu, Mingzheng; Wang, Shufang; Sun, Yue; Zhang, Gufeng; Shi, Jianrong
2016-08-02
The present study was performed to identify prevailing Fusarium species and the environmental factors affecting their frequencies and the contamination of grain with major mycotoxins in Jiangsu province. The precipitation levels were 184.2mm, 156.4mm, and 245.8mm in the years 2013-2015, respectively, and the temperature fluctuated by an average of 10.6±7.2°C in 2013, 10.9±7.2°C in 2014, and 10.6±6.3°C in 2015. Co-occurrence of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3ADON), and 15-acetyldeoxynivalenol (15ADON) were observed in wheat. The average concentrations of DON were 879.3±1127.8, 627.8±640.5, and 1628.6±2,168.0μg/kg in 2013-2015, respectively. The average concentrations of 3ADON were 43.5±59.0, 71.2±102.5, and 33.5±111.9μg/kg in 2013-2015, respectively. We found that the average concentration of DON in wheat was positively correlated with precipitation (r=0.998, p<0.01), and 3ADON was negatively correlated with precipitation (r=-0.887, p<0.05). However, there was no correlation between precipitation and 15ADON or nivalenol (NIV). The differences in temperature were not as significant as the differences in rainfall amount over a short time period. Therefore, there were no correlations between temperature and the concentrations of trichothecenes, excluding 3ADON (r=0.996, p<0.01). Our data indicated that Fusarium asiaticum is the primary pathogenic fungus prevalent in the Fusarium head blight disease nursery. The trichothecene chemotype composition differed between Fusarium graminearum sensu stricto (s. str.) and F. asiaticum isolates. The 3ADON chemotype was found only among strains of F. asiaticum. The NIV chemotype was not observed among strains of F. graminearum, while the 15ADON chemotype represented 100% of the F. graminearum strains collected. The results of this study indicated no correlations between environmental conditions and the species or genetic chemotype composition of pathogens in Jiangsu province in 2013-2015. Copyright © 2016 Elsevier B.V. All rights reserved.
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Wu, Wenda; Zhou, Hui-Ren; Pan, Xiao; Pestka, James J
Trichothecene mycotoxins, potent translational inhibitors that are associated with human food poisonings and damp-building illnesses, are of considerable concern to animal and human health. Food refusal is a hallmark of exposure of experimental animals to deoxynivalenol (DON) and other Type B trichothecenes but less is known about the anorectic effects of foodborne Type A trichothecenes (e.g., T-2 toxin, HT-2 toxin), airborne Type D trichothecenes (e.g. satratoxin G [SG]) or functionally analogous metabolites that impair protein synthesis. Here, we utilized a well-described mouse model of food intake to compare the anorectic potencies of T-2 toxin, HT-2 toxin, and SG to that of emetine, a medicinal alkaloid derived from ipecac that inhibits translation. Intraperitoneal (IP) administration with T-2 toxin, HT-2 toxin, emetine and SG evoked anorectic responses that occurred within 0.5 h that lasted up to 96, 96, 3 and 96 h, respectively, with lowest observed adverse effect levels (LOAELs) being 0.1, 0.1, 2.5 and 0.25 mg/kg BW, respectively. When delivered via natural routes of exposure, T-2 toxin, HT-2 toxin, emetine (oral) and SG (intranasal) induced anorectic responses that lasted up to 48, 48, 3 and 6 h, respectively with LOAELs being 0.1, 0.1, 0.25, and 0.5 mg/kg BW, respectively. All four compounds were generally much more potent than DON which was previously observed to have LOAELs of 1 and 2.5 mg/kg BW after IP and oral dosing, respectively. Taken together, these anorectic potency data will be valuable in discerning the relative risks from trichothecenes and other translational inhibitors of natural origin.
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Vanheule, Adriaan; Audenaert, Kris; De Boevre, Marthe; Landschoot, Sofie; Bekaert, Boris; Munaut, Françoise; Eeckhout, Mia; Höfte, Monica; De Saeger, Sarah; Haesaert, Geert
2014-07-02
Global food safety depends on continuous monitoring of food contaminants such as mycotoxins in cereals and cereal-derived products. Here, we combine this type of investigation with quantitative occurrence data on Fusarium infestation of these products in extensive correlation studies. Finally, this contributes to a thorough understanding of the presence, origin and physiology of Fusarium Head Blight (FHB) related mycotoxins and the correlations within their ranks. Two hundred and thirty-seven samples were analyzed from diverse cereal matrices, representing the most important stages of the cereal food and feed chain in Belgium. Food, feed and non-processed field samples were investigated, with a strong emphasis on whole-grain food products. Two approaches were pursued to estimate the full scope of FHB and its repercussions: UPLC-MS/MS was applied to detect twelve different mycotoxins, and Q-PCR was used to measure the presence of ten Fusarium species. We found that different matrices have different characteristic contamination profiles, and extensive correlation studies identified certain mycotoxins for future assessment (e.g. moniliformin produced by the Fusarium avenaceum/Fusarium tricinctum species group). The investigated harvest year of 2012 yielded many non-processed field materials containing elevated levels of deoxynivalenol (DON), while even in a so-called DON-year less prevalent toxins such as T-2 and HT-2 might be considered problematic due to their consistent co-occurrence with related mycotoxins. Our data illustrate complex interactions between the many Fusarium species that are responsible for FHB and their mycotoxins. Correlation studies demonstrate that consistent co-occurrence of mycotoxins is not to be neglected, and pinpoint issues for future surveillance and legislation. Copyright © 2014 Elsevier B.V. All rights reserved.
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Albonico, Marco; Schütz, Luis F; Caloni, Francesca; Cortinovis, Cristina; Spicer, Leon J
2016-08-01
There is now overwhelming evidence of global contamination of commodities with Fusarium mycotoxins. Fumonisin B1 (FB1) is a Fusarium mycotoxin frequently occurring in corn in combination with deoxynivalenol (DON), α-zearalenol (α-ZEA) and β-zearalenol (β-ZEA). The aim of this study was to determine if FB1, alone and combined with DON or α-ZEA or β-ZEA, can affect cell proliferation and steroid production of bovine granulosa cells (GC). A species-specific model with bovine granulosa cells (GC) was used to study the potential endocrine disruptor effects of FB1 alone and in co-exposure. In the presence of β-ZEA (30 ng/mL), FB1 at 30 ng/mL showed a stimulatory effect on GC numbers. Insulin-like growth factor-1 (IGF1)-stimulated cell proliferation was decreased after exposure to β-ZEA alone at 5.0 μg/mL and FB1 with α-ZEA and β-ZEA at the same concentration. Regarding steroid production, FB1 at 30 ng/mL and 100 ng/mL amplified the inhibitory effect of β-ZEA (30 ng/mL) on estradiol (E2) production, while FB1 alone increased (P < 0.05) IGF1-induced E2 production. α-ZEA alone decreased (P < 0.05) E2 production, whereas β-ZEA alone and in combination with FB1 decreased (P < 0.05) E2 production. These studies indicate for the first time that the Fusarium mycotoxin FB1 along with other mycotoxins can affect GC proliferation and steroid production, which ultimately could influence reproductive function in cattle. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Endocrine activity of mycotoxins and mycotoxin mixtures.
Demaegdt, Heidi; Daminet, Britt; Evrard, Annick; Scippo, Marie-Louise; Muller, Marc; Pussemier, Luc; Callebaut, Alfons; Vandermeiren, Karine
2016-10-01
Reporter gene assays incorporating nuclear receptors (estrogen, androgen, thyroid β and PPARγ2) have been implemented to assess the endocrine activity of 13 mycotoxins and their mixtures. As expected, zearalenone and its metabolites α-zearalenol and β- zearalenol turned out to have the strongest estrogenic potency (EC50 8,7 10-10 ± 0,8; 3,1 10-11 ± 0,5 and 1,3 10-8 ± 0,3 M respectively). The metabolite of deoxynivalenol, 3-acetyl-deoxynivalenol also had estrogenic activity (EC50 3,8 10-7 ± 1,1 M). Furthermore, most of the mycotoxins (and their mixtures) showed anti-androgenic effects (15-acetyldeoxynivalenol, 3-acetyl-deoxynivalenol and α-zearalenol with potencies within one order of magnitude of that of the reference compound flutamide). In particular, deoxynivalenol and 15-acetyl-deoxynivalenol acted as antagonists for the PPARy2 receptor. When testing mixtures of mycotoxins on the same cell systems, we showed that most of the mixtures reacted as predicted by the concentration addition (CA) theory. Generally, the CA was within the 95% confidence interval of the observed ones, only minor deviations were detected. Although these reporter gene tests cannot be directly extrapolated in vivo, they can be the basis for further research. Especially the additive effects of ZEN and its metabolites are of importance and could have repercussions in vivo. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Giese, Henriette; Sondergaard, Teis Esben; Sørensen, Jens Laurids
2013-01-01
Growth conditions are known to affect the production of secondary metabolites in filamentous fungi. The influence of different nitrogen sources and the transcription factor AreA on the production of mycotoxins in Fusarium graminearum was examined. Growth on glutamine or NH4-sources was poor and asparagine was found to be a preferential nitrogen source for F. graminearum. Deletion of areA led to poor growth on NaNO₃ suggesting its involvement in regulation of the nitrate reduction process. In addition utilization of aspartic acid, histidine, isoleucine, leucine, threonine, tyrosine, and valine as nitrogen sources was shown to depend of a functional AreA. AreA was shown to be required for the production of the mycotoxins deoxynivalenol (DON), zearalenone, and fusarielin H regardless of the nutrient medium. Deletion of nmr, the repressor of AreA under nitrogen sufficient conditions, had little effect on either growth or toxin production. AreA appears to regulate production of some mycotoxins directly or indirectly independent on nitrogen status and plays a role in utilization of certain amino acids. Copyright © 2013 The British Mycological Society. All rights reserved.
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Climate change increases deoxynivalenol contamination of wheat in north-western Europe.
van der Fels-Klerx, H J; Olesen, J E; Madsen, M S; Goedhart, P W
2012-01-01
Climate change will affect the development of cereal crops and the occurrence of mycotoxins in these crops, but so far little research has been done on quantifying the expected effects. The aim of this study was to assess climate change impacts on the occurrence of deoxynivalenol in wheat grown in north-western Europe by 2040, considering the combined effects of shifts in wheat phenology and climate. The study used climate model data for the future period of 2031-2050 relative to the baseline period of 1975-1994. A weather generator was used for generating synthetic series of daily weather data for both the baseline and the future periods. Available models for wheat phenology and prediction of deoxynivalenol concentrations in north-western Europe were used. Both models were run for winter wheat and spring wheat, separately. The results showed that both flowering and full maturation of wheat will be earlier in the season because of climate change effects, about 1 to 2 weeks. Deoxynivalenol contamination was found to increase in most of the study region, with an increase of the original concentrations by up to 3 times. The study results may inform governmental and industrial risk managers to underpin decision-making and planning processes in north-western Europe. On the local level, deoxynivalenol contamination should be closely monitored to pick out wheat batches with excess levels at the right time. Using predictive models on a more local scale could be helpful to assist other monitoring measures to safeguard food safety in the wheat supply chain.
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Hallier, Arnaud; Celette, Florian; Coutarel, Julie; David, Christophe
2013-01-01
Fusarium head blight caused by different varieties of Fusarium species is one of the major serious worldwide diseases found in wheat production. It is therefore important to be able to quantify the deoxynivalenol concentration in wheat. Unfortunately, in mycotoxin quantification, due to the uneven distribution of mycotoxins within the initial lot, it is difficult, or even impossible, to obtain a truly representative analytical sample. In previous work we showed that the sampling step most responsible for variability was grain sampling. In this paper, it is more particularly the step scaling down from a laboratory sample of some kilograms to an analytical sample of a few grams that is investigated. The naturally contaminated wheat lot was obtained from an organic field located in the southeast of France (Rhône-Alpes) from the year 2008-2009 cropping season. The deoxynivalenol level was found to be 50.6 ± 2.3 ng g⁻¹. Deoxynivalenol was extracted with a acetonitrile-water mix and quantified by gas chromatography-electron capture detection (GC-ECD). Three different grain sampling techniques were tested to obtain analytical samples: a technique based on manually homogenisation and division, a second technique based on the use of a rotating shaker and a third on the use of compressed air. Both the rotating shaker and the compressed air techniques enabled a homogeneous laboratory sample to be obtained, from which representative analytical samples could be taken. Moreover, the techniques did away with many repetitions and grinding. This study, therefore, contributes to sampling variability reduction in the evaluation of deoxynivalenol contamination of organic wheat grain, and then, at a reasonable cost.
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Chilaka, Cynthia Adaku; De Boevre, Marthe; Atanda, Olusegun Oladimeji; De Saeger, Sarah
2018-05-03
The stability of the Fusarium mycotoxins fumonisin B 1 , deoxynivalenol, T-2 toxin, and zearalenone during processing of Nigerian traditional spices (dawadawa, okpehe, and ogiri) and beer (burukutu) using artificially contaminated raw materials was investigated. Results revealed the reduction of these toxins in all the final products. Boiling played a significant role (p < 0.05) in Fusarium mycotoxin reduction in the traditional spices. The highest percentage reduction of deoxynivalenol (76%) and zearalenone (74%) was observed during okpehe processing (boiled for 12 h). Dehulling and fermentation further demonstrated a positive influence on the reduction of these toxins with a total reduction ranging from 85 to 98% for dawadawa, 86 to 100% for okpehe, and 57 to 81% for ogiri. This trend was also observed during the production of traditional beer (burukutu), with malting and brewing playing a major impact in observed reduction. In addition, other metabolites including deoxynivalenol-3-glucoside, 15-acetyl-deoxynivalenol, α-zearalenol, and β-zearalenol which were initially not present in the raw sorghum were detected in the final beer product at the following concentrations 26 ± 11, 16 ± 7.7, 22 ± 18, and 31 ± 16 μg/kg, respectively. HT-2 toxin was also detected at a concentration of 36 ± 13 μg/kg along the processing chain (milled malted fraction) of the traditional beer. For the traditional spices, HT-2 toxin was detected (12 μg/kg) in ogiri. Although there was a reduction of mycotoxins during processing, appreciable concentrations of these toxins were still detected in the final products. Thus, the use of good quality raw materials significantly reduces mycotoxin contamination in final products.
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Bánáti, Hajnalka; Darvas, Béla; Fehér-Tóth, Szilvia; Czéh, Árpád; Székács, András
2017-02-22
Levels of mycotoxins produced by Fusarium species in genetically modified (GM) and near-isogenic maize, were determined using multi-analyte, microbead-based flow immunocytometry with fluorescence detection, for the parallel quantitative determination of fumonisin B1, deoxynivalenol, zearalenone, T-2, ochratoxin A, and aflatoxin B1. Maize varieties included the genetic events MON 810 and DAS-59122-7 , and their isogenic counterparts. Cobs were artificially infested by F. verticillioides and F. proliferatum conidia, and contained F. graminearum and F. sporotrichoides natural infestation. The production of fumonisin B1 and deoxynivalenol was substantially affected in GM maize lines: F. verticillioides , with the addition of F. graminearum and F. sporotrichoides , produced significantly lower levels of fumonisin B1 (~300 mg·kg -1 ) in DAS-59122-7 than in its isogenic line (~580 mg·kg -1 ), while F. proliferatum , in addition to F. graminearum and F. sporotrichoides , produced significantly higher levels of deoxynivalenol (~18 mg·kg -1 ) in MON 810 than in its isogenic line (~5 mg·kg -1 ). Fusarium verticillioides , with F. graminearum and F. sporotrichoides , produced lower amounts of deoxynivalenol and zearalenone than F. proliferatum , with F. graminearum and F. sporotrichoides . T-2 toxin production remained unchanged when considering the maize variety. The results demonstrate the utility of the Fungi-Plex™ quantitative flow immunocytometry method, applied for the high throughput parallel determination of the target mycotoxins.
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Osselaere, A; Devreese, M; Watteyn, A; Vandenbroucke, V; Goossens, J; Hautekiet, V; Eeckhout, M; De Saeger, S; De Baere, S; De Backer, P; Croubels, S
2012-08-01
Contamination of feeds with mycotoxins is a worldwide problem and mycotoxin-detoxifying agents are used to decrease their negative effect. The European Food Safety Authority recently stated guidelines and end-points for the efficacy testing of detoxifiers. Our study revealed that plasma concentrations of deoxynivalenol and deepoxy-deoxynivalenol were too low to assess efficacy of 2 commercially available mycotoxin-detoxifying agents against deoxynivalenol after 3 wk of continuous feeding of this mycotoxin at concentrations of 2.44±0.70 mg/kg of feed and 7.54±2.20 mg/kg of feed in broilers. This correlates with the poor absorption of deoxynivalenol in poultry. A safety study with 2 commercially available detoxifying agents and veterinary drugs showed innovative results with regard to the pharmacokinetics of 2 antibiotics after oral dosing in the drinking water. The plasma and kidney tissue concentrations of oxytetracycline were significantly higher in broilers receiving a biotransforming agent in the feed compared with control birds. For amoxicillin, the plasma concentrations were significantly higher for broilers receiving an adsorbing agent in comparison to birds receiving the biotransforming agent, but not to the control group. Mycotoxin-detoxifying agents can thus interact with the oral bioavailability of antibiotics depending on the antibiotic and detoxifying agent, with possible adverse effects on the health of animals and humans.
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Jovaišienė, J; Bakutis, B; Baliukonienė, V; Gerulis, G
2016-01-01
One hundred two samples of feeds made in Lithuania, which included maize silage, grass-legume silage, hay and ensiled crimped maize were investigated during 2008-2012 for contamination with some mycotoxins. The highest concentrations of mycotoxins determined were those of deoxynivalenol (DON)--471.0 μg/kg and aflatoxin B1 (AFB1)--21.2 μg/kg in ryegrass silage from bales, and zearalenone (ZEA)--625.0 μg/kg in maize silage from trenches. The present study has been carried out based on these data because animal feeds contaminated with mycotoxins can cause reduced productivity of dairy cows and health disorders in the long term. The aim of this study was to investigate the long-term exposure of toxic effects of a diet naturally contaminated with low concentrations of mycotoxins on milk composition and biochemical, hematological, immunological parameters of dairy cows and to determine the anti-mycotoxin effect of Mycofix Plus 3.E. Twenty eight clinically healthy, medium productive Lithuanian Red cows were selected. ZEA was a major contaminant found in the corn silage at concentration levels of up to 1000.0 μg/kg of dry matter. DON was the second major found in the hay at concentration levels of up to 600.0 μg/kg of dry matter. The highest concentration AFB1- 10.0 μg/kg was determined in ground barley. The Anti-Mycotoxin Additive (AMA) Mycofix Plus 3.E was given individually to 14 cows at a concentration of 40.0 g daily for 9 weeks. The present results indicate that feeds naturally contaminated with low concentration of mycotoxins produced by Fusarium spp. and Aspergillus spp. in a diet of dairy cows can have a negative influence on somatic cell count, blood parameters and immunity. The addition of an Anti-Mycotoxin Additive (Mycofix Plus 3.E) to diet of dairy cows can prevent many of these effects.
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Updated survey of Fusarium species and toxins in Finnish cereal grains.
Hietaniemi, Veli; Rämö, Sari; Yli-Mattila, Tapani; Jestoi, Marika; Peltonen, Sari; Kartio, Mirja; Sieviläinen, Elina; Koivisto, Tauno; Parikka, Päivi
2016-05-01
The aim of the project was to produce updated information during 2005-14 on the Fusarium species found in Finnish cereal grains, and the toxins produced by them, as the last comprehensive survey study of Fusarium species and their toxins in Finland was carried out at the turn of the 1960s and the 1970s. Another aim was to use the latest molecular and chemical methods to investigate the occurrence and correlation of Fusarium species and their mycotoxins in Finland. The most common Fusarium species found in Finland in the FinMyco project 2005 and 2006 were F. avenaceum, F. culmorum, F. graminearum, F. poae, F. sporotrichioides and F. langsethiae. F. avenaceum was the most dominant species in barley, spring wheat and oat samples. The occurrence of F. culmorum and F. graminearum was high in oats and barley. Infection by Fusarium fungi was the lowest in winter cereal grains. The incidence of Fusarium species in 2005 was much higher than in 2006 due to weather conditions. F. langsethiae has become much more common in Finland since 2001. F. graminearum has also risen in the order of importance. A highly significant correlation was found between Fusarium graminearum DNA and deoxynivalenol (DON) levels in Finnish oats, barley and wheat. When comparing the FinMyco data in 2005-06 with the results of the Finnish safety monitoring programme for 2005-14, spring cereals were noted as being more susceptible to infection by Fusarium fungi and the formation of toxins. The contents of T-2 and HT-2 toxins and the frequency of exceptionally high DON concentrations all increased in Finland during 2005-14. Beauvericin (BEA), enniatins (ENNs) and moniliformin (MON) were also very common contaminants of Finnish grains in 2005-06. Climate change is leading to warmer weather, and this may indicate more changes in Finnish Fusarium mycobiota and toxin contents and profiles in the near future.
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Immunomodulatory effects of feed-borne Fusarium mycotoxins in chickens infected with coccidia.
Girgis, George N; Sharif, Shayan; Barta, John R; Boermans, Herman J; Smith, Trevor K
2008-11-01
The potential for Fusarium mycotoxins to modulate immunity was studied in chickens raised to 10 weeks of age using an enteric coccidial infection model. Experimental diets included: control, diets containing grains naturally contaminated with Fusarium mycotoxins, and diets containing contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent (GMA). Contaminated diets contained up to 3.8 microg/g deoxynivalenol (DON), 0.3 microg/g 15-acetyl DON and 0.2 microg/g zearalenone. An optimized mixture (inducing lesions without mortality) of Eimeria acervulina, E. maxima and E. tenella was used to challenge birds at 8 weeks of age. Immune parameters were studied prior to challenge, at the end of the challenge period (7 days post-inoculation, PI), and at the end of the recovery period (14 days PI). Total serum immunoglobulin (Ig) A and IgG concentrations in challenged birds fed the contaminated diet were higher than controls at the end of the challenge period. Serum concentration of IgA, but not IgG, was significantly decreased at the end of the recovery period in birds fed the contaminated diet. The percentage of CD4+ and CD8+ cell populations in blood mononuclear cells decreased significantly at the end of the challenge period in birds fed the control or the contaminated diet compared to their percentages prior to challenge. The pre-challenge percentage of CD8+ population was restored at the end of the recovery period only in birds fed the control diet. Interferon-gamma (IFN-gamma) gene expression in caecal tonsils was up-regulated in challenged birds fed the contaminated diet at the end of the challenge period. No significant effect of diet was observed on oocyst counts despite the changes in the studied immune parameters. It was concluded that Fusarium mycotoxins modulate the avian immune system. This modulation involves alteration of gene expression but apparently does not enhance susceptibility or resistance to a primary coccidial challenge.
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Revajová, Viera; Levkut, Mikuláš; Levkutová, Mária; Bořutová, Radka; Grešaková, Lubomíra; Košiková, Božena; Leng, Lubomír
2013-09-01
The objective of the study was to investigate the effects of lignin supplementation of a diet contaminated with the Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEA) on peripheral blood leukocytes and duodenal immunocompetent cells in broiler chickens. From day 1 after hatching, all chickens were fed an identical control diet for two weeks. Then chickens of Group 1 continued to be fed the control diet, whereas Group 2 was fed the same diet supplemented with lignin at 0.5% level. Simultaneously, Group 3 started to receive a diet contaminated with DON (2.95 mg kg-1) and ZEA (1.59 mg kg-1), while Group 4 received an identical contaminated diet supplemented with 0.5% lignin for further two weeks. Samples of blood and duodenal tissue were collected from 6 birds of each group at 4 weeks of age. Neither counts of white blood cells nor phagocytic function in the peripheral blood were significantly affected in the mycotoxin- and/or lignin-treated birds. As compared to the control, increased numbers of IgM-bearing cells were found in the peripheral blood in Group 3 fed the contaminated diet (P < 0.05) and in Group 4 given the contaminated diet supplemented with lignin (P < 0.01). While the contaminated diet led to reduced numbers of duodenal CD4+ cells, in Group 2 treated only with lignin the number of duodenal CD4+ cells was increased. Lignin enrichment of the contaminated diet did not eliminate the mycotoxin-induced reduction in the number of duodenal CD4+ cells. The results suggest that dietary supplementation of lignin as an indigestible compound to poultry feed may increase the density of some intestinal immunocompetent cells without exerting effects on that in the peripheral blood. However, when added to a diet contaminated with Fusarium mycotoxins, lignin did not prevent the mycotoxin-induced changes in the numbers of blood and intestinal immunocompetent cells.
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Beccari, Giovanni; Prodi, Antonio; Tini, Francesco; Bonciarelli, Umberto; Onofri, Andrea; Oueslati, Souheib; Limayma, Marwa; Covarelli, Lorenzo
2017-01-01
In this study, conducted for three years on eleven malting barley varieties cultivated in central Italy, the incidence of different mycotoxigenic fungal genera, the identification of the Fusarium species associated with the Fusarium Head Blight (FHB) complex, and kernels contamination with deoxynivalenol (DON) and T-2 mycotoxins were determined. The influence of climatic conditions on Fusarium infections and FHB complex composition was also investigated. Fusarium species were always present in the three years and the high average and maximum temperatures during anthesis mainly favored their occurrence. The FHB complex was subject to changes during the three years and the main causal agents were F. poae, F. avenaceum, F. tricinctum and F. graminearum, which, even if constantly present, never represented the principal FHB agent. The relative incidence of Fusarium species changed because of climatic conditions occurring during the seasons. The FHB complex was composed of many different Fusarium species and some of them were associated with a specific variety and/or with specific weather parameters, indicating that the interaction between a certain plant genotype and climatic conditions may influence the presence of Fusarium spp. causing infections. With regard to mycotoxin contamination, T-2 toxin, in some cases, was found in kernels at levels that exceeded EU recommended values. PMID:28353653
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Trichothecene mycotoxins and their determinants in settled dust related to grain production.
Nordby, Karl-Christian; Halstensen, Anne Straumfors; Elen, Oleif; Clasen, Per-Erik; Langseth, Wenche; Kristensen, Petter; Eduard, Wijnand
2004-01-01
We hypothesise that inhalant exposure to mycotoxins causes developmental outcomes and certain hormone-related cancers that are associated with grain farming in an epidemiological study. The aim of the present study was to identify and validate determinants of measured trichothecene mycotoxins in grain dust as work environmental trichothecene exposure indicators. Settled grain dust was collected in 92 Norwegian farms during seasons of 1999 and 2000. Production characteristics and climatic data were studied as determinants of trichothecenes in settled dust samples obtained during the production of barley (N = 59), oats (N = 32), and spring wheat (N = 13). Median concentrations of trichothecenes in grain dust were <20, 54, and < 50 mg/kg (ranges < 20-340, < 30-2400, and < 50-1200) for deoxynivalenol (DON), HT-2 toxin (HT-2) and T-2 toxin (T-2) respectively. Late blight potato rot (fungal) forecasts have been broadcast in Norway to help prevent this potato disease. Fungal forecasts representing wet, temperate, and humid meteorological conditions were identified as strong determinants of trichothecene mycotoxins in settled grain dust in this study. Differences in cereal species, production properties and districts contributed less to explain mycotoxin concentrations. Fungal forecasts are validated as indicators of mycotoxin exposure of grain farmers and their use in epidemiological studies may be warranted.
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Swamy, H V L N; Smith, T K; Cotter, P F; Boermans, H J; Sefton, A E
2002-07-01
Three hundred sixty, 1-d-old male broiler chicks were fed diets containing grains naturally contaminated with Fusarium mycotoxins for 56 d. The four diets included control (0.14 mg/kg deoxynivalenol, 18 mg/ kg fusaric acid, < 0.1 mg/kg zearalenone), low level of contaminated grains (4.7 mg/kg deoxynivalenol, 20.6 mg/kg fusaric acid, 0.2 mg/kg zearalenone), and high level of contaminated grains without (8.2 mg/kg deoxynivalenol, 20.3 mg/kg fusaric acid, 0.56 mg/kg zearalenone) and with (9.7 mg/kg deoxynivalenol, 21.6 mg/kg fusaric acid, 0.8 mg/kg zearalenone) 0.2% esterified-glucomannan polymer derived from Saccharomyces cerevisiae1026 (E-GM). Body weight gain and feed consumption responded in a significant quadratic fashion to the inclusion of contaminated grains during the finisher period. Efficiency of feed utilization, however, was not affected by diets. The feeding of contaminated grains in the finisher period also caused significant linear increases in blood erythrocyte count and serum uric acid concentration and a significant linear decline in the serum lipase activity. Dietary inclusion of contaminated grains resulted in a significant quadratic effect on serum albumin and y-glutamyltransferase activity. Blood hemoglobin and biliary IgA concentrations, however, responded in significant linear and quadratic fashions. Supplementation of E-GM counteracted most of the blood parameter alterations caused by the Fusarium mycotoxin-contaminated grains and reduced breast muscle redness. It was concluded that broiler chickens may be susceptible to Fusarium mycotoxicoses when naturally contaminated grains are fed containing a combination of mycotoxins.
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Bernhoft, A.; Torp, M.; Clasen, P.-E.; Løes, A.-K.; Kristoffersen, A.B.
2012-01-01
A total of 602 samples of organically and conventionally grown barley, oats and wheat was collected at grain harvest during 2002–2004 in Norway. Organic and conventional samples were comparable pairs regarding cereal species, growing site and harvest time, and were analysed for Fusarium mould and mycotoxins. Agronomic and climatic factors explained 10–30% of the variation in Fusarium species and mycotoxins. Significantly lower Fusarium infestation and concentrations of important mycotoxins were found in the organic cereals. The mycotoxins deoxynivalenol (DON) and HT-2 toxin (HT-2) constitute the main risk for human and animal health in Norwegian cereals. The impacts of various agronomic and climatic factors on DON and HT-2 as well as on their main producers F. graminearum and F. langsethiae and on total Fusarium were tested by multivariate statistics. Crop rotation with non-cereals was found to reduce all investigated characteristics significantly – mycotoxin concentrations as well as various Fusarium infestations. No use of mineral fertilisers and herbicides was also found to decrease F. graminearum, whereas lodged fields increased the occurrence of this species. No use of herbicides was also found to decrease F. langsethiae, but for this species the occurrence was lower in lodged fields. Total Fusarium infestation was decreased with no use of fungicides or mineral fertilisers, and with crop rotation, as well as by using herbicides and increased by lodged fields. Clay and to some extent silty soils seemed to reduce F. graminearum in comparison with sandy soils. Concerning climate factors, low temperature before grain harvest was found to increase DON; and high air humidity before harvest to increase HT-2. F. graminearum was negatively correlated with precipitation in July but correlated with air humidity before harvest. F. langsethiae was correlated with temperature in July. Total Fusarium increased with increasing precipitation in July. Organic cereal farmers have fewer cereal intense rotations than conventional farmers. Further, organic farmers do not apply mineral fertiliser or pesticides (fungicides, herbicides or insecticides), and have less problem with lodged fields. The study showed that these agronomic factors were related to the infestation of Fusarium species and the concentration of mycotoxins. Hence, it is reasonable to conclude that farming system (organic versus conventional) impacts Fusarium infestation, and that organic management tends to reduce Fusarium and mycotoxins. However, Fusarium infestation and mycotoxin concentrations may be influenced by a range of factors not studied here, such as local topography and more local climate, as well as cereal species and variety. PMID:22494553
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Fate of trichothecene mycotoxins during the processing: milling and baking.
Lancova, K; Hajslova, J; Kostelanska, M; Kohoutkova, J; Nedelnik, J; Moravcova, H; Vanova, M
2008-05-01
Toxic secondary metabolites produced by fungi representing Fusarium genus are common contaminants in cereals worldwide. To estimate the dietary intake of these trichothecene mycotoxins, information on their fate during cereal processing is needed. Up-to-date techniques such as high-performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (LC-MS/MS) was used for the analysis of seven trichothecenes (deoxynivalenol, nivalenol, HT-2 toxin, T-2 toxin, 15- and 3-acetyldeoxynivalenol, and fusarenon-X) in bread production chain (wheat grains, intermediate products collected during milling and baking process, breads). Regardless of whether the grains were naturally infected or artificially inoculated by Fusarium spp. in the field, the fractions obtained from the grain-cleaning procedure contained the highest mycotoxin levels. During milling the highest concentrations of deoxynivalenol were found in the bran, the lowest in the reduction flours. Baking at 210 degrees C for 14 min had no significant effect on deoxynivalenol levels. The rheological properties of dough measured by fermentograph, maturograph, oven rise recorder, and laboratory baking test were carried out, and based on the obtained results the influence of mycotoxin content on rheological behaviour was investigated.
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Nathanail, Alexis V; Gibson, Brian; Han, Li; Peltonen, Kimmo; Ollilainen, Velimatti; Jestoi, Marika; Laitila, Arja
2016-07-15
An investigation was conducted to determine the fate of deoxynivalenol, deoxynivalenol-3-glucoside, HT-2 toxin and T-2 toxin, during a four-day fermentation with the lager yeast Saccharomyces pastorianus. The influence of excessive mycotoxin concentrations on yeast growth, productivity and viability were also assessed. Mycotoxins were dosed at varying concentrations to 11.5° Plato wort. Analysis of yeast revealed that presence of the toxins even at concentrations up to 10,000 μg/L had little or no effect on sugar utilisation, alcohol production, pH, yeast growth or cell viability. Of the dosed toxin amounts 9-34% were removed by the end of fermentation, due to physical binding and/or biotransformation by yeast. Deoxynivalenol-3-glucoside was not reverted to its toxic precursor during fermentation. Processing of full-scan liquid chromatography-quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS) data with MetaboLynx and subsequent LC-QTOF-MS/MS measurements resulted in annotation of several putative metabolites. De(acetylation), glucosylation and sulfonation were the main metabolic pathways activated. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Kolpin, Dana W.; Hoerger, Corinne C.; Meyer, Michael T.; Wettstein, Felix E.; Hubbard, Laura E.; Bucheli, Thomas D.
2010-01-01
This study provides the first broad-scale investigation on the spatial and temporal occurrence of phytoestrogens and mycotoxins in streams in the United States. Fifteen stream sites across Iowa were sampled five times throughout the 2008 growing season to capture a range of climatic and crop-growth conditions. Basin size upstream from sampling sites ranged from 7 km2 to >836,000 km2 Atrazine (herbicide) also was measured in all samples as a frame-of-reference agriculturally derived contaminant. Target compounds were frequently detected in stream samples: atrazine (100%), formononetin (80%), equol (45%), deoxynivalenol (43%), daidzein (32%), biochanin A (23%), zearalenone (13%), and genistein (11%). The nearly ubiquitous detection of formononetin (isoflavone) suggests a widespread agricultural source, as one would expect with the intense row crop and livestock production present across Iowa. Conversely, the less spatially widespread detections of deoxynivalenol (mycotoxin) suggest a more variable source due to the required combination of proper host and proper temperature and moisture conditions necessary to promote Fusarium spp. infections. Although atrazine concentrations commonly exceeded 100 ng L-1 (42/75 measurements), only deoxynivalenol (6/56 measurements) had concentrations that occasionally exceeded this level. Temporal patterns in concentrations varied substantially between atrazine, formononetin, and deoxynivalenol, as one would expect for contaminants with different source inputs and processes of formation and degradation. The greatest phytoestrogen and mycotoxin concentrations were observed during spring snowmelt conditions. Phytoestrogens and mycotoxins were detected at all sampling sites regardless of basin size. The ecotoxicological effects from long-term, low-level exposures to phytoestrogens and mycotoxins or complex chemicals mixtures including these compounds that commonly take place in surface water are poorly understood and have yet to be systematically investigated in environmental studies.
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Kolpin, D.W.; Hoerger, C.C.; Meyer, M.T.; Wettstein, F.E.; Hubbard, L.E.; Bucheli, T.D.
2010-01-01
This study provides the first broad-scale investigation on the spatial and temporal occurrence of phytoestrogens and mycotoxins in streams in the United States. Fifteen stream sites across Iowa were sampled five times throughout the 2008 growing season to capture a range of climatic and crop-growth conditions. Basin size upstream from sampling sites ranged from 7 km2 to >836,000 km2. Atrazine (herbicide) also was measured in all samples as a frame-ofreference agriculturally derived contaminant. Target compounds were frequently detected in stream samples: atrazine (100%), formononetin (80%), equol (45%), deoxynivalenol (43%), daidzein (32%), biochanin A (23%), zearalenone (13%), and genistein (11%). Th e nearly ubiquitous detection of formononetin (isoflavone) suggests a widespread agricultural source, as one would expect with the intense row crop and livestock production present across Iowa. Conversely, the less spatially widespread detections of deoxynivalenol (mycotoxin) suggest a more variable source due to the required combination of proper host and proper temperature and moisture conditions necessary to promote Fusarium spp. infections. Although atrazine concentrations commonly exceeded 100 ng L-1 (42/75 measurements), only deoxynivalenol (6/56 measurements) had concentrations that occasionally exceeded this level. Temporal patterns in concentrations varied substantially between atrazine, formononetin, and deoxynivalenol, as one would expect for contaminants with different source inputs and processes of formation and degradation. The greatest phytoestrogen and mycotoxin concentrations were observed during spring snowmelt conditions. Phytoestrogens and mycotoxins were detected at all sampling sites regardless of basin size. The ecotoxicological effects from long-term, low-level exposures to phytoestrogens and mycotoxins or complex chemicals mixtures including these compounds that commonly take place in surface water are poorly understood and have yet to be systematically investigated in environmental studies. Copyright ?? 2010 by the American Society of Agronomy.
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Selwet, Marek
2011-01-01
The objective of the performed investigations was to isolate pathogenic fungi from contaminated maize cobs, to assess the appearance of maize cob fusariosis and to determine grain contamination with deoxynivalenol in the cultivation of genetically modified maize containing a gene resistance against European corn borer (Ostrinia nubilalis Hbn) as well as selected non-modified cultivars. The plant material comprised the following genetically modified maize cultivar: DKC 3421 YG (MON 810) and non-modified cultivars obtained from Smolice Plant Breeding Ltd., IHAR Group: Junak (FAO 210-220), Prosna (FAO 220), SMH (FAO 230), Baca (FAO 220). Prior to harvesting, the occurrence of maize cob fusariosis was determined in the 89 (BBCH) developmental ripening stage. Microbiological assessment was carried out on grains selected from cobs characterized by various pathological symptoms. In 2008, a total of 133 isolates was obtained from the examined samples of infected maize plants, of which 51 isolates were species-identified, while in 2009, the total of 123 isolates were determined, of which 63 were species-identified. In both experimental years, the majority of isolates contained fungi from the Fusarium genus. The performed analysis of mean levels of cob contamination by fusarioses revealed that DKC 3421 YG (MON 810) and SMH (FAO 230) cultivars showed the smallest levels of contamination as well as the lowest percent of cob contamination per plant, while Junak (FAO 210-220) and Baca (FAO 220) cultivars were characterized by the highest degree of contamination. The lowest deoxynivalenol concentrations were determined in years 2008 and 2009 in the case of the DKC 3421 YG (MON 810) cultivar, whereas Prosna (FAO 220) cultivar was characterized by the highest deoxynivalenol concentration.
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Xu, Jiao-Jiao; Zhou, Jian; Huang, Bai-Fen; Cai, Zeng-Xuan; Xu, Xiao-Min; Ren, Yi-Ping
2016-06-01
A simple and reliable method of ultra high performance liquid chromatography coupled with photo-diode array detection has been proposed for the simultaneous determination of deoxynivalenol and its acetylated derivatives in wheat flour and rice, especially focusing on the optimization of sample extraction, cleanup, and chromatographic separation conditions. Sample pretreatment consisted of a first step using a quick, easy, cheap, effective, rugged, and safe based extraction procedure and a subsequent cleanup step based on solid-phase extraction. The method was extensively validated in wheat flour and rice, obtaining satisfactory analytical performance with good linearity (R(2) ≥ 0.999), acceptable recoveries (80.0-104.4%), and repeatability (RSDs 1.3-10.7%). The limits of detection (21.7-57.4 μg/kg) and quantitation (72.3-191.4 μg/kg) for deoxynivalenols were lower than those usually permitted by various countries' legislation in these food matrices. The method was applied to 34 wheat and rice samples. The results were further compared with results of ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Fate of Fusarium Toxins during Brewing.
Habler, Katharina; Geissinger, Cajetan; Hofer, Katharina; Schüler, Jan; Moghari, Sarah; Hess, Michael; Gastl, Martina; Rychlik, Michael
2017-01-11
Some information is available about the fate of Fusarium toxins during the brewing process, but only little is known about the single processing steps in detail. In our study we produced beer from two different barley cultivars inoculated with three different Fusarium species, namely, Fusarium culmorum, Fusarium sporotrichioides, and Fusarium avenaceum, producing a wide range of mycotoxins such as type B trichothecenes, type A trichothecenes, and enniatins. By the use of multi-mycotoxin LC-MS/MS stable isotope dilution methods we were able to follow the fate of Fusarium toxins during the entire brewing process. In particular, the type B trichothecenes deoxynivalenol, 3-acetyldeoxynivalenol, and 15-acetyldeoxynivalenol showed similar behaviors. Between 35 and 52% of those toxins remained in the beer after filtration. The contents of the potentially hazardous deoxynivalenol-3-glucoside and the type A trichothecenes increased during mashing, but a rapid decrease of deoxynivalenol-3-glucoside content was found during the following steps of lautering and wort boiling. The concentration of enniatins greatly decreased with the discarding of spent grains or finally with the hot break. The results of our study show the retention of diverse Fusarium toxins during the brewing process and allow for assessing the food safety of beer regarding the monitored Fusarium mycotoxins.
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Toxigenic Fusarium spp. as determinants of trichothecene mycotoxins in settled grain dust.
Halstensen, Anne Straumfors; Nordby, Karl-Christian; Klemsdal, Sonja Sletner; Elen, Oleif; Clasen, Per-Erik; Eduard, Wijnand
2006-12-01
Trichothecenes are immunosuppressive mycotoxins produced mainly by Fusarium spp. and often are detected as natural contaminants of grain and other agricultural products. Exposure to trichothecenes through inhalation during grain work may represent possible health risks for grain farmers. We aimed, therefore, to investigate the level of Fusarium spp. and trichothecenes in settled grain dust collected during work on 92 Norwegian farms. Mycotoxins were determined by gas chromatography-mass spectrometry, whereas the Fusarium spp. were identified and quantified both by species-specific semiquantitative polymerase chain reaction (PCR) and by cultivation. All potential trichothecene-producing molds in the grain dust were quantified using a PCR assay specific for tri5, the gene coding for trichodiene synthase that catalyzes the first step in the trichothecene biosynthesis. We performed correlation analysis between mold-DNA and mycotoxins to assess whether the PCR-detected DNA could be used as indicators of the mycotoxins. The methodological problem of detecting small amounts of airborne mycotoxins during grain work may then be avoided. Whereas the trichothecene-producing Fusarium species in grain dust could not be identified or quantified to a sufficient extent by cultivation, all investigated Fusarium spp. could be specifically detected by PCR and quantified from the DNA agarose gel band intensities. Furthermore, we observed a strong correlation between the trichothecenes HT-2 toxin (HT-2) or T-2 toxin (T-2) and DNA specific for tri5 (r = 0.68 for HT-2 and r = 0.50 for T-2; p < 0.001), F. langsethiae (r = 0.77 for HT-2 and r = 0.59 for T-2; p < 0.001), or F. poae (r = 0.41 for HT-2 and r = 0.35 for T-2; p < 0.001). However, only a moderate correlation was observed between the trichothecene deoxynivalenol (DON) and the combination of its producers, F. culmorum and F. graminearum (r = 0.24, p = 0.02), and no significant correlation was observed between DON and tri5. PCR clearly improved the detection of toxigenic Fusaria as potential sources of health risks for farmers inhaling grain dust during work, but the use of Fusarium-DNA as indicators for trichothecenes should be used cautiously.
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Schmeitzl, Clemens; Varga, Elisabeth; Warth, Benedikt; Kugler, Karl G.; Malachová, Alexandra; Michlmayr, Herbert; Wiesenberger, Gerlinde; Mayer, Klaus F. X.; Mewes, Hans-Werner; Krska, Rudolf; Schuhmacher, Rainer; Berthiller, Franz; Adam, Gerhard
2015-01-01
Increasing frequencies of 3-acetyl-deoxynivalenol (3-ADON)-producing strains of Fusarium graminearum (3-ADON chemotype) have been reported in North America and Asia. 3-ADON is nearly nontoxic at the level of the ribosomal target and has to be deacetylated to cause inhibition of protein biosynthesis. Plant cells can efficiently remove the acetyl groups of 3-ADON, but the underlying genes are yet unknown. We therefore performed a study of the family of candidate carboxylesterases (CXE) genes of the monocot model plant Brachypodium distachyon. We report the identification and characterization of the first plant enzymes responsible for deacetylation of trichothecene toxins. The product of the BdCXE29 gene efficiently deacetylates T-2 toxin to HT-2 toxin, NX-2 to NX-3, both 3-ADON and 15-acetyl-deoxynivalenol (15-ADON) into deoxynivalenol and, to a lesser degree, also fusarenon X into nivalenol. The BdCXE52 esterase showed lower activity than BdCXE29 when expressed in yeast and accepts 3-ADON, NX-2, 15-ADON and, to a limited extent, fusarenon X as substrates. Expression of these Brachypodium genes in yeast increases the toxicity of 3-ADON, suggesting that highly similar genes existing in crop plants may act as susceptibility factors in Fusarium head blight disease. PMID:26712789
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Proximal sensing of within-field mycotoxin variation - a case study in Northeast Germany
NASA Astrophysics Data System (ADS)
Mueller, Marina; Koszinski, Sylvia; Bangs, Donovan E.; Wehrhan, Marc; Ullrich, Andreas; Verch, Gernot; Brenning, Alexander
2017-04-01
Fusarium head blight is a global problem in agriculture that results in yield losses and, more seriously, produces harmful toxins that enter the food chain. This study (Müller et al. 2016) builds on previous research identifying within-field humidity as an important factor in infection processes by Fusarium fungi and its mycotoxin production. Environmental variables describing topographic control of humidity (topographic wetness index TWI), soil texture and related moisture by electrical conductivity (ECa), and canopy humidity by density (normalized difference vegetation index NDVI) were explored in their relationship to the fungal infection rates and mycotoxin accumulation. Field studies at four sites in NE German Lowlands were performed in 2009 and 2011. Sites differed slightly in soil textural properties and, more pronounced, mean annual precipitation. Sampling positions were selected by usage of NDVI values range from remote sensing data base. Environmental data included elevation and its derivatives like topographic wetness index (TWI) from a DEM25, electrical conductivity distribution maps (5 x 5 m) based on EM38DD survey and, orthorectified RapidEye imagery (5 x 5 m2) with resulting NDVI distributions across the field sites. Grain yield, fungal infection rate, microbiological characteristics and mycotoxin accumulation were determined at 223 field positions. Statistical analysis incorporated Spearman rank order correlations and three regression methods (censored regression models, linear mixed-effects models and spatial linear mixed-effects models). Kriging was used to visualize the spatial patterns and trends. All analyses were performed by R software. In 2011, a more wet year than 2009, high Fusarium infection rates and a high concentration of mycotoxins were stated, the latter once exceeding EU threshold values. For both years associations between NDVI and microbiological variables were found, but being more pronounced and more often significant for 2011 than for 2009. ECa was only related with deoxynivalenol concentration (DON) and abundance of trichothecene-producing fusaria (tri6 gene copy number) in 2009 and, to DON and zearalenone (ZEA) in 2011. In contrast to former findings no correlations were found between TWI and mycological data. NDVI and, less importantly, ECa were essential predictors in all three regression models. Mycotoxins DON and ZEA distribution maps could be interpolated by kriging with internal drift based on these two proximal predictor variables. Providing spatial patterns of mycotoxigenic fungi and its effects may be used to infer mycotoxin hot spots, to develop models for risk assessment and, to manage plant and crop treatments or even harvest. Müller, M.E.H., Koszinski, S., Bangs, D.E. et al. Precision Agric (2016) 17: 698. doi:10.1007/s11119-016-9444-y
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Chu, F.S.
1988-05-12
1) A new direct ELISA, which involves the use of a generic type antibody for T-2 toxin and 3-acetyl-neosolaniol-hemisuccinate (3-Ac-NEOS-HS) conjugated to horseradish peroxidase for the analysis of T-2 toxin metabolites, was established. 2) Analysis of 5 metabolites, i.e., T-2 toxin, HT-2, T-2-4ol, 3'-OH-HT-2, NEOS and a mixture of these five toxins, added at toxin concentrations of 50 and 200 ng/mL of urine, revealed that 87% of the added toxins were recovered analytically at both concentrations in the ELISA, and 69% in RIA and the 200 ppb level. 3) A series of monkey urine samples obtained from a metabolic studymore » was subjected to the new ELISA protocol was as well as to RIA. 4) An extensive study to evaluate a fluorometric method, i.e., hit-and-run assay was made. 5) A direct ELISA for deoxynivalenol (DON) was established. 6) Further improvements for the ELISA for saxitoxin were made. 7) New approaches to produce antibody against nivalenol and saxitoxin were made, but the antibody titers were low. 8) Antibody against microcystin (MCS) was obtained from rabbits after immunizing the animals with MCS-BSA conjugate. 9) A chemical method for the preparation of different monoacetoxyscirpenol (MAS) was established. 10) Routine preparation of different immunochemical reagents continued. Phytotoxins Biological warfare agents; Dinoflagellates. (AW)« less
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Abia, Wilfred A; Warth, Benedikt; Ezekiel, Chibundu N; Sarkanj, Bojan; Turner, Paul C; Marko, Doris; Krska, Rudolf; Sulyok, Michael
2017-09-01
Toxins of microbial origin frequently contaminate foodstuffs worldwide and pose a serious hazard to humans. This study reports on LCMS/MS quantification of multiple fungal and bacterial toxins, from household sampling of 50 traditionally prepared maize-fufu samples from Bamunka village, western highlands of Cameroon. Seventy-four metabolites including aflatoxin B1 (AFB1) (12/50: mean 0.9, range n.d-1.8 μg kg -1 ), cereulide (50/50: mean 37; range 1-236 μg kg -1 ), deoxynivalenol (DON) (50/50: mean 23, range 14-55 μg kg -1 ), fumonisin B1 (FB1) (50/50: mean: 151, range 48-709 μg kg -1 ), nivalenol (NIV) (50/50; mean 268, range 116-372 μg kg -1 ), patulin (PAT) (15/50:mean 105, range 12-890 μg kg -1 ) and zearalenone (ZEN) (50/50: mean 49, range 5-150) were detected; and of note every sample contained at least 27 toxic compounds. While individual toxin levels were mostly low there is always concern regarding mixtures, for which data are absent or limited. This study reports several novel observations of toxins not previously reported in maize, and the mixture of toxins, e.g. cereulide, PAT and ZEN derivatives (ZEN-cis and ZENsulfate-cis) are reported for the first time in Cameroonian food. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.
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Current Status of Mycotoxin Contamination of Food Commodities in Zimbabwe.
Nleya, Nancy; Adetunji, Modupeade Christianah; Mwanza, Mulunda
2018-05-03
Agricultural products, especially cereal grains, serve as staple foods in sub-Saharan Africa. However, climatic conditions in this region can lead to contamination of these commodities by moulds, with subsequent production of mycotoxins posing health risks to both humans and animals. There is limited documentation on the occurrence of mycotoxins in sub-Saharan African countries, leading to the exposure of their populations to a wide variety of mycotoxins through consumption of contaminated foods. This review aims at highlighting the current status of mycotoxin contamination of food products in Zimbabwe and recommended strategies of reducing this problem. Zimbabwe is one of the African countries with very little information with regards to mycotoxin contamination of its food commodities, both on the market and at household levels. Even though evidence of multitoxin occurrence in some food commodities such as maize and other staple foods exist, available published research focuses only on Aspergillus and Fusarium mycotoxins, namely aflatoxins, deoxynivalenol (DON), trichothecenes, fumonisins, and zearalenone (ZEA). Occurrence of mycotoxins in the food chain has been mainly associated with poor agricultural practices. Analysis of mycotoxins has been done mainly using chromatographic and immunological methods. Zimbabwe has adopted European standards, but the legislation is quite flexible, with testing for mycotoxin contamination in food commodities being done voluntarily or upon request. Therefore, the country needs to tighten its legislation as well as adopt stricter standards that will improve the food safety and security of the masses.
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Döll, Katharina; Karlovsky, Petr; Deising, Holger B.; Wirsel, Stefan G. R.
2013-01-01
Fusarium graminearum is a plant pathogen infecting several important cereals, resulting in substantial yield losses and mycotoxin contamination of the grain. Triazole fungicides are used to control diseases caused by this fungus on a worldwide scale. Our previous microarray study indicated that 15 ABC transporter genes were transcriptionally upregulated in response to tebuconazole treatment. Here, we deleted four ABC transporter genes in two genetic backgrounds of F. graminearum representing the DON (deoxynivalenol) and the NIV (nivalenol) trichothecene chemotypes. Deletion of FgABC3 and FgABC4 belonging to group I of ABC-G and to group V of ABC-C subfamilies of ABC transporters, respectively, considerably increased the sensitivity to the class I sterol biosynthesis inhibitors triazoles and fenarimol. Such effects were specific since they did not occur with any other fungicide class tested. Assessing the contribution of the four ABC transporters to virulence of F. graminearum revealed that, irrespective of their chemotypes, deletion mutants of FgABC1 (ABC-C subfamily group V) and FgABC3 were impeded in virulence on wheat, barley and maize. Phylogenetic context and analyses of mycotoxin production suggests that FgABC3 may encode a transporter protecting the fungus from host-derived antifungal molecules. In contrast, FgABC1 may encode a transporter responsible for the secretion of fungal secondary metabolites alleviating defence of the host. Our results show that ABC transporters play important and diverse roles in both fungicide resistance and pathogenesis of F. graminearum. PMID:24244413
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Early response of wheat antioxidant system with special reference to Fusarium head blight stress.
Spanic, Valentina; Viljevac Vuletic, Marija; Abicic, Ivan; Marcek, Tihana
2017-06-01
Fusarium head blight (FHB) is a destructive fungal disease of wheat (Triticum aestivum L.) that causes significant grain yield losses and end-use quality reduction associated with contamination by the mycotoxin deoxynivalenol (DON). Three winter wheat varieties ('Vulkan', 'Kraljica' and 'Golubica') were screened for FHB resistance using artificial inoculation technique under field conditions. The aim of this study was to examine a relationship between FHB resistance and the effectiveness of enzyme antioxidant system of wheat varieties under different sampling times (3, 15, 24, 48, 96, 120 and 336 hai). In the time-course experiments FHB-resistant variety 'Vulkan' showed rapid induction of ascorbate peroxidase (APX) and polyphenol oxidase (PPO) activity in the early stages after infection (3 hai) and it seems that in 'Vulkan' FHB-resistance is associated with antioxidative enzymes activity. Moderately FHB resistant variety 'Kraljica' showed the higher guaiacol peroxidase (POD) activity and higher H 2 O 2 content after 24 hai, increased malondialdehyde (MDA) content at the beginning of infection (3, 15 hai) while induction of catalase (CAT), APX and PPO was delayed. FHB-susceptible variety 'Golubica' involved antioxidant enzymes in defense response much later. Based on our results the activity of antioxidant enzymes (APX and PPO) was more pronounced in 'Vulkan' than in FHB-medium resistant variety 'Kraljica' and FHB-susceptible 'Golubica'. The differences in antioxidant response of wheat varieties under Fusarium infestation could be the result of genetic properties. Copyright © 2017 Elsevier Masson SAS. All rights reserved.
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Antioxidant agents against trichothecenes: new hints for oxidative stress treatment
Nepovimova, Eugenie; Wang, Yun; Yang, Hualin; Li, Li; Zhang, Xiujuan; Kuca, Kamil
2017-01-01
Trichothecenes are a group of mycotoxins mainly produced by fungi of genus Fusarium. Due to high toxicity and widespread dissemination, T-2 toxin and deoxynivalenol (DON) are considered to be the most important compounds of this class. Trichothecenes generate free radicals, including reactive oxygen species (ROS), which induce lipid peroxidation, decrease levels of antioxidant enzymes, and ultimately lead to apoptosis. Consequently, oxidative stress is an active area of research on the toxic mechanisms of trichothecenes, and identification of antioxidant agents that could be used against trichothecenes is crucial for human health. Numerous natural compounds have been analyzed and have shown to function very effectively as antioxidants against trichothecenes. In this review, we summarize the molecular mechanisms underlying oxidative stress induced by these compounds, and discuss current knowledge regarding such antioxidant agents as vitamins, quercetin, selenium, glucomannan, nucleotides, antimicrobial peptides, bacteria, polyunsaturated fatty acids, oligosaccharides, and plant extracts. These products inhibit trichothecene-induced oxidative stress by (1) inhibiting ROS generation and induced DNA damage and lipid peroxidation; (2) increasing antioxidant enzyme activity; (3) blocking the MAPK and NF-κB signaling pathways; (4) inhibiting caspase activity and apoptosis; (5) protecting mitochondria; and (6) regulating anti-inflammatory actions. Finally, we summarize some decontamination methods, including bacterial and yeast biotransformation and degradation, as well as mycotoxin-binding agents. This review provides a comprehensive overview of antioxidant agents against trichothecenes and casts new light on the attenuation of oxidative stress. PMID:29299181
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Malachova, Alexandra; Dzuman, Zbynek; Veprikova, Zdenka; Vaclavikova, Marta; Zachariasova, Milena; Hajslova, Jana
2011-12-28
Fusarium toxins, Alternaria toxins, and ergot alkaloids represent common groups of mycotoxins that can be found in cereals grown under temperate climatic conditions. Because most of them are chemically and thermally stable, these toxic fungal secondary metabolites might be transferred from grains into the final products. To get information on the commensurate contamination of various cereal-based products collected from the Czech retail market in 2010, the occurrence of "traditional" mycotoxins such as groups of A and B trichothecenes and zearalenone, less routinely determined Alternaria toxins (alternariol, alternariol monomethyl ether and altenuene), ergot alkaloids (ergosine, ergocryptine, ergocristine, and ergocornine) and "emerging" mycotoxins (enniatins A, A1, B, and B1 and beauvericin) were monitored. In a total 116 samples derived from white flour and mixed flour, breakfast cereals, snacks, and flour, only trichothecenes A and B and enniatins were found. Deoxynivalenol was detected in 75% of samples with concentrations ranging from 13 to 594 μg/kg, but its masked form, deoxynivalenol-3-β-d-glucoside, has an even higher incidence of 80% of samples, and concentrations ranging between 5 and 72 μg/kg were detected. Nivalenol was found only in three samples at levels of 30 μg/kg. For enniatins, all of the samples investigated were contaminated with at least one of four target enniatins. Enniatin A was detected in 97% of samples (concentration range of 20-2532 μg/kg) followed by enniatin B with an incidence in 91% of the samples (concentration range of 13-941 μg/kg) and enniatin B1 with an incidence of 80% in the samples tested (concentration range of 8-785 μg/kg). Enniatin A1 was found only in 44% of samples at levels ranging between 8 and 851 μg/kg.
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Data Analyses and Modelling for Risk Based Monitoring of Mycotoxins in Animal Feed
van der Fels-Klerx, H.J. (Ine); Adamse, Paulien; Punt, Ans; van Asselt, Esther D.
2018-01-01
Following legislation, European Member States should have multi-annual control programs for contaminants, such as for mycotoxins, in feed and food. These programs need to be risk based implying the checks are regular and proportional to the estimated risk for animal and human health. This study aimed to prioritize feed products in the Netherlands for deoxynivalenol and aflatoxin B1 monitoring. Historical mycotoxin monitoring results from the period 2007–2016 were combined with data from other sources. Based on occurrence, groundnuts had high priority for aflatoxin B1 monitoring; some feed materials (maize and maize products and several oil seed products) and complete/complementary feed excluding dairy cattle and young animals had medium priority; and all other animal feeds and feed materials had low priority. For deoxynivalenol, maize by-products had a high priority, complete and complementary feed for pigs had a medium priority and all other feed and feed materials a low priority. Also including health consequence estimations showed that feed materials that ranked highest for aflatoxin B1 included sunflower seed and palmkernel expeller/extracts and maize. For deoxynivalenol, maize products were ranked highest, followed by various small grain cereals (products); all other feed materials were of lower concern. Results of this study have proven to be useful in setting up the annual risk based control program for mycotoxins in animal feed and feed materials. PMID:29373559
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Sharma, Pallavi; Gangola, Manu P; Huang, Chen; Kutcher, H Randy; Ganeshan, Seedhabadee; Chibbar, Ravindra N
2018-01-01
An in vitro spike culture method was optimized to evaluate Fusarium head blight (FHB) resistance in wheat (Triticum aestivum) and used to screen a population of ethyl methane sulfonate treated spike culture-derived variants (SCDV). Of the 134 SCDV evaluated, the disease severity score of 47 of the variants was ≤30%. Single nucleotide polymorphisms (SNP) in the UDP-glucosyltransferase (UGT) genes, TaUGT-2B, TaUGT-3B, and TaUGT-EST, differed between AC Nanda (an FHB-susceptible wheat variety) and Sumai-3 (an FHB-resistant wheat cultivar). SNP at 450 and 1,558 bp from the translation initiation site in TaUGT-2B and TaUGT-3B, respectively were negatively correlated with FHB severity in the SCDV population, whereas the SNP in TaUGT-EST was not associated with FHB severity. Fusarium graminearum strain M7-07-1 induced early expression of TaUGT-2B and TaUGT-3B in FHB-resistant SCDV lines, which were associated with deoxynivalenol accumulation and reduced FHB disease progression. At 8 days after inoculation, deoxynivalenol concentration varied from 767 ppm in FHB-resistant variants to 2,576 ppm in FHB-susceptible variants. The FHB-resistant SCDV identified can be used as new sources of FHB resistance in wheat improvement programs.
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Reinhold, Lilli; Reinhardt, Katja
2011-05-01
In this presentation, the mycotoxin levels-as analysed by the analytical centre for mycotoxin surveillance of the state food laboratory (LAVES Braunschweig)-for approximately 500 food samples are reported. The samples were collected in the year 2009 at retail in the German federal state of Lower Saxony. Aflatoxin and ochratoxin A were analysed in dried fruits, spices, cereals and tree nuts. Ochratoxin A was detected in all samples of dried vine fruits, at levels up to 8.1 μg/kg. Aflatoxins and ochratoxin A were also found in nutmeg and curry powder: the maximum regulatory levels for aflatoxins were exceeded in 25% of the nutmeg samples. Nearly all samples of basmati rice contained aflatoxins, although at levels below the maximum regulatory level in all but one sample. Aflatoxins were also detected in about 50% of hazelnut samples, in 20% of the samples the maximum levels was exceeded (maximum 23.2 μg/kg). In contrast, aflatoxin contents in pistachios were surprisingly low. Fusarium toxins were analysed in cereals and cereal products such as flour, bread, and pasta. Deoxynivalenol (DON) was the predominant toxin found in these samples: DON was found in about 40% of the samples, although the maximum levels were not exceeded (max. 418 μg/kg). Fumonisins (FBs) and zearalenone (ZEA) were specifically analysed in maize products (snacks, flour and oil). Most of these samples (80%) were positive, but at levels not exceeding the maximum levels. Maximum levels were 98 μg/kg (ZEA) and 577 μg/kg (sum of FB1 and FB2). Ergot alkaloids (six major alkaloids) were analysed in rye flour, and approximately 50% were positive. The highest concentration of ergot alkaloids was 1,063 μg/kg; the predominant alkaloids were ergotamine and ergocristine. In conclusion, the results indicate that continuous and efficient control measures for mycotoxins in a wide range of critical foods are necessary to ensure compliance with maximum levels. Although the mycotoxin levels in the vast majority of samples were below maximum levels, year-to-year variation and changes in the production of relevant commodities may result in a different picture in the future.
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Munawar, Hasim; Smolinska-Kempisty, Katarzyna; Cruz, Alvaro Garcia; Canfarotta, Francesco; Piletska, Elena; Karim, Khalku; Piletsky, Sergey A
2018-06-20
The enzyme-linked immunosorbent assay (ELISA) has been used as a standard tool for monitoring food and animal feed contamination from the carcinogenic fumonisin B1 (FB1). Unfortunately, ELISA is not always efficient due to the instability of the antibody and enzyme components in the immunoassay, the presence of natural enzyme inhibitors in the samples and the high levels of non-specific protein binding. Additionally, the production of antibodies for ELISA can be time-consuming and costly, due to the involvement of animals in the manufacturing process. To overcome these limiting factors, a molecularly imprinted nanoparticle based assay (MINA) has been developed, where the molecularly imprinted nanoparticles (nanoMIPs) replace the primary antibody used in a competitive ELISA. Herein, computational modelling was used to design the nanoMIPs by selecting monomers that specifically interact with FB1. The affinity of the monomers to FB1 was verified by measuring their binding in affinity chromatography experiments. The nanoMIPs were produced by solid phase synthesis and the results showed that nanoMIPs had a hydrodynamic diameter of around 249 ± 29 nm. The assay tested in model samples is highly selective and does not show cross-reactivity with other mycotoxins such as fumonisin B2 (FB2), aflatoxin B1 (AFB1), citrinin (CTT), zearalenone (ZEA), and deoxynivalenol (DON). The MINA allows the detection of FB1 in the concentration range of 10 pM-10 nM with a detection limit of 1.9 pM and a recovery of 108.13-113.76%.
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Kos, Gregor; Sieger, Markus; McMullin, David; Zahradnik, Celine; Sulyok, Michael; Öner, Tuba; Mizaikoff, Boris; Krska, Rudolf
2016-10-01
The rapid identification of mycotoxins such as deoxynivalenol and aflatoxin B 1 in agricultural commodities is an ongoing concern for food importers and processors. While sophisticated chromatography-based methods are well established for regulatory testing by food safety authorities, few techniques exist to provide a rapid assessment for traders. This study advances the development of a mid-infrared spectroscopic method, recording spectra with little sample preparation. Spectral data were classified using a bootstrap-aggregated (bagged) decision tree method, evaluating the protein and carbohydrate absorption regions of the spectrum. The method was able to classify 79% of 110 maize samples at the European Union regulatory limit for deoxynivalenol of 1750 µg kg -1 and, for the first time, 77% of 92 peanut samples at 8 µg kg -1 of aflatoxin B 1 . A subset model revealed a dependency on variety and type of fungal infection. The employed CRC and SBL maize varieties could be pooled in the model with a reduction of classification accuracy from 90% to 79%. Samples infected with Fusarium verticillioides were removed, leaving samples infected with F. graminearum and F. culmorum in the dataset improving classification accuracy from 73% to 79%. A 500 µg kg -1 classification threshold for deoxynivalenol in maize performed even better with 85% accuracy. This is assumed to be due to a larger number of samples around the threshold increasing representativity. Comparison with established principal component analysis classification, which consistently showed overlapping clusters, confirmed the superior performance of bagged decision tree classification.
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Suppressive Effect of Trichoderma spp. on toxigenic Fusarium species.
Błaszczyk, Lidia; Basińska-Barczak, Aneta; Ćwiek-Kupczyńska, Hanna; Gromadzka, Karolina; Popiel, Delfina; Stępień, Łukasz
2017-03-30
The aim of the present study was to examine the abilities of twenty-four isolates belonging to ten different Trichoderma species (i.e., Trichoderma atroviride, Trichoderma citrinoviride, Trichoderma cremeum, Trichoderma hamatum, Trichoderma harzianum, Trichoderma koningiopsis, Trichoderma longibrachiatum, Trichoderma longipile, Trichoderma viride and Trichoderma viridescens) to inhibit the mycelial growth and mycotoxin production by five Fusarium strains (i.e., Fusarium avenaceum, Fusarium cerealis, Fusarium culmorum, Fusarium graminearum and Fusarium temperatum). Dual-culture bioassay on potato dextrose agar (PDA) medium clearly documented that all of the Trichoderma strains used in the study were capable of influencing the mycelial growth of at least four of all five Fusarium species on the fourth day after co-inoculation, when there was the first apparent physical contact between antagonist and pathogen. The qualitative evaluation of the interaction between the colonies after 14 days of co-culturing on PDA medium showed that ten Trichoderma strains completely overgrew and sporulated on the colony at least one of the tested Fusarium species. Whereas, the microscopic assay provided evidence that only T. atroviride AN240 and T. viride AN255 formed dense coils around the hyphae of the pathogen from where penetration took place. Of all screened Trichoderma strains, T. atroviride AN240 was also found to be the most efficient (69-100% toxin reduction) suppressors of mycotoxins (deoxynivalenol, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, nivalenol, zearalenone, beauvericin, moniliformin) production by all five Fusarium species on solid substrates. This research suggests that T. atroviride AN240 can be a promising candidate for the biological control of toxigenic Fusarium species.
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Spanjer, Martien C; Rensen, Peter M; Scholten, Jos M
2008-04-01
Mycotoxin analysis is usually carried out by high performance liquid chromatography after immunoaffinity column cleanup or in enzyme-linked immunosorbent assay tests. These methods normally involve determination of single compounds only. EU legislation already exists for the aflatoxins, ochratoxin A and patulin in food, and legislation will come into force for deoxynivalenol, zearalenone and the fumonisins in 2007. To enforce the various legal limits, it would be preferable to determine all mycotoxins by routine analysis in different types of matrices in one single extract. This would also be advantageous for HACCP control purposes. For this reason, a multi-method was developed with which 33 mycotoxins in various products could be analysed simultaneously. The mycotoxins were extracted with an acetonitrile/water mixture, diluted with water and then directly injected into a LC-MS/MS system. The mycotoxins were separated by reversed-phase HPLC and detected using an electrospray ionisation interface (ESI) and tandem MS, using MRM in the positive ion mode, to increase specificity for quality control. The following mycotoxins could be analysed in a single 30-min run: Aflatoxins B1, B2, G1 and G2, ochratoxin A, deoxynivalenol, zearalenone, T-2 toxin, HT-2 toxin, alpha-zearalenol, alpha-zearalanol, beta-zearalanol, sterigmatocystin, cyclopiazonic acid, penicillic acid, fumonisins B1, B2 and B3, diacetoxyscirpenol, 3- and 15-acetyl-deoxynivalenol, zearalanone, ergotamin, ergocornin, ergocristin, alpha-ergocryptin, citrinin, roquefortin C, fusarenone X, nivalenol, mycophenolic acid, alternariol and alternariol monomethyl ether. The limit of quantification for the aflatoxins and ochratoxin A was 1.0 microg kg(-1) and for deoxynivalenol 50 microg kg(-1). The quantification limits for the other mycotoxins were in the range 10-200 microg kg(-1). The matrix effect and validation data are presented for between 13 and 24 mycotoxins in peanuts, pistachios, wheat, maize, cornflakes, raisins and figs. The method has been compared with the official EU method for the determination of aflatoxins in food and relevant FAPAS rounds. The multi-mycotoxin method has been proven by the detection of more than one mycotoxin in maize, buckwheat, figs and nuts. The LC-MS/MS technique has also been applied to baby food, which is subject to lower limits for aflatoxin B1 and ochratoxin A, ergot alkaloids in naturally contaminated rye and freeze-dried silage samples.
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Liu, Chun-Sen; Sun, Chun-Xiao; Tian, Jia-Yue; Wang, Zhuo-Wei; Ji, Hong-Fei; Song, Ying-Pan; Zhang, Shuai; Zhang, Zhi-Hong; He, Ling-Hao; Du, Miao
2017-05-15
Two unique immunosensors made of aluminum-based metal-organic frameworks (MOFs), namely, 515- and 516-MOFs, with 4,4',4''-nitrilotribenzoic acid (H 3 NTB) were successfully obtained to efficiently assess food safety. The as-prepared 515- and 516-MOFs exhibited superior thermal and physicochemical stability, high electrochemical activity, and good biocompatibility. Among these immunosensors, 516-MOF showed a preferable biosensing ability toward analytes determined by electrochemical techniques. The developed 516-MOF-based electrochemical biosensor not only demonstrated high sensitivity with low detection limits of 0.70 and 0.40pgmL -1 toward vomitoxin and salbutamol, respectively, but also showed good selectivity in the presence of other interferences. Therefore, with the advantages of high sensitivity, good selectivity, and simple operation, this new strategy is believed to exhibit great potential for simple and convenient detection of poisonous and harmful residues in food. Copyright © 2017 Elsevier B.V. All rights reserved.
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Ribeiro, Cláudia M R; Maia, Alexandra S; Ribeiro, Ana R; Couto, Cristina; Almeida, Agostinho A; Santos, Mariana; Tiritan, Maria E
2016-10-14
Natural organic compounds such as phytoestrogens and phytosterols found in various plants, as well as mycotoxins produced by fungi, can be found in aquatic environments. The aim of this study was to investigate the occurrence of three different classes of natural estrogenic compounds, i.e., phytoestrogens, phytosterols and mycotoxins, in estuarine water samples from the Ave River estuary. For that, water samples were collected at five sampling points distributed along the estuary at low tide, during 1 year, processed by solid-phase extraction (SPE) and analyzed by gas chromatography coupled to mass spectrometry (GC-MS). To correlate the presence of phytoestrogens and phytosterols in the estuarine water, local flora was collected on riverside. Trace elements content and physicochemical parameters such as nutrients and dissolved oxygen were also determined seasonally at each sampling point, to give insights for the evaluation of water quality and anthropogenic pressure. Both phytoestrogens and phytosterols showed a seasonal variation, with the highest values observed in spring and summer and the lowest in winter. Daidzein (DAID) was found up to 404.0 ng L(-1) in spring and coumestrol (COUM) was found up to 165.0 ng L(-1) in summer. The mycotoxin deoxynivalenol (DON) was ubiquitously determined with values ranging from 59.5 to 642.4 ng L(-1). Nutrients and metals distribution and concentration varied among sampling stations and seasons. This study revealed for the first time the presence of mycotoxins, various classes of phytoestrogens and stigmasterol (STG) in estuarine water from the Ave River (Portugal), and the evaluation of the water quality confirmed that this estuary is still highly impacted by anthropogenic activities.
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Antonissen, Gunther; Devreese, Mathias; De Baere, Siegrid; Martel, An; Van Immerseel, Filip; Croubels, Siska
2017-03-01
Cytochrome P450 (CYP450) drug biotransformation enzymes and multidrug resistance (MDR) proteins may influence drug disposition processes. The first part of the study aimed to evaluate the effect of mycotoxins deoxynivalenol (DON) and/or fumonisins (FBs), at contamination levels approaching European Union guidance levels, on intestinal and hepatic CYP450 enzymes and MDR proteins gene expression in broiler chickens. mRNA expression of genes encoding CYP450 enzymes (CYP3A37, CYP1A4 and CYP1A5) and drug transporters (MDR1/ABCB1 and MRP2/ABCC2) was determined using qRT-PCR. A significant up-regulation of CYP1A4 (P = 0.037) and MDR1 (P = 0.036) was observed in the jejunum of chickens fed a diet contaminated with FBs. The second part of this study aimed to investigate the impact of feeding a FBs contaminated diet on the oral absorption of enrofloxacin (10 mg/kg BW), a MDR1 substrate. A significant (P = 0.045), however small, decreased area under the plasma concentration-time curve (AUC 0-48 h, mean ± SD) was observed for enrofloxacin in chickens fed the FBs contaminated diet compared to the control group, 16.28 ± 1.82 h μg/mL versus 18.27 ± 1.79 h μg/mL. These findings suggest that concurrent administration of drugs with FBs contaminated feed might alter the pharmacokinetic characteristics of CYP1A4 substrate drugs and MDR1 substrates, such as enrofloxacin. Copyright © 2017 Elsevier Ltd. All rights reserved.
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Detection of deoxynivalenol using biolayer interferometry
USDA-ARS?s Scientific Manuscript database
Biolayer interferometry allows for the real time monitoring of the interactions between molecules without the need for reagents with enzymatic, fluorescent, or radioactive labels. The technology is based upon the changes in interference pattern of light reflected from the surface of an optical fiber...
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Maresca, Marc; Yahi, Nouara; Younes-Sakr, Lama
Mycotoxins are fungal secondary metabolites responsible of food-mediated intoxication in animals and humans. Deoxynivalenol, ochratoxin A and patulin are the best known enteropathogenic mycotoxins able to alter intestinal functions resulting in malnutrition, diarrhea, vomiting and intestinal inflammation in vivo. Although their effects on intestinal barrier and transport activities have been extensively characterized, the mechanisms responsible for their pro-inflammatory effect are still poorly understood. Here we investigated if mycotoxin-induced intestinal inflammation results from a direct and/or indirect pro-inflammatory activity of these mycotoxins on human intestinal epithelial cells, using differentiated Caco-2 cells as model and interleukin 8 (IL-8) as an indicator ofmore » intestinal inflammation. Deoxynivalenol was the only mycotoxin able to directly increase IL-8 secretion (10- to 15-fold increase). We also investigated if these mycotoxins could indirectly stimulate IL-8 secretion through: (i) a modulation of the action of pro-inflammatory molecules such as the interleukin-1beta (IL-1{beta}), and/or (ii) an increase in the transepithelial passage of non-invasive commensal Escherichia coli. We found that deoxynivalenol, ochratoxin A and patulin all potentiated the effect of IL-1{beta} on IL-8 secretion (ranging from 35% to 138% increase) and increased the transepithelial passage of commensal bacteria (ranging from 12- to 1544-fold increase). In addition to potentially exacerbate established intestinal inflammation, these mycotoxins may thus participate in the induction of sepsis and intestinal inflammation in vivo. Taken together, our results suggest that the pro-inflammatory activity of enteropathogenic mycotoxins is mediated by both direct and indirect effects.« less
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NASA Astrophysics Data System (ADS)
Meyer-Wolfarth, Friederike; Schrader, Stefan; Oldenburg, Elisabeth; Brunotte, Joachim; Weinert, Joachim
2017-04-01
In agroecosystems soil-borne fungal plant diseases are major yield-limiting factors which are difficult to control. Fungal plant pathogens, like Fusarium species, survive as a saprophyte in infected tissue like crop residues and endanger the health of the following crop by increasing the infection risk for specific plant diseases. In infected plant organs, these pathogens are able to produce mycotoxins. Mycotoxins like deoxynivalenol (DON) persist during storage, are heat resistant and of major concern for human and animal health after consumption of contaminated food and feed, respectively. Among fungivorous soil organisms, there are representatives of the soil fauna which are obviously antagonistic to a Fusarium infection and the contamination with mycotoxins. Specific members of the soil macro-, meso-, and microfauna provide a wide range of ecosystem services including the stimulation of decomposition processes which may result in the regulation of plant pathogens and the degradation of environmental contaminants. Investigations under laboratory conditions and in field were conducted to assess the functional linkage between soil faunal communities and plant pathogenic fungi (Fusarium culmorum). The aim was to examine if Fusarium biomass and the content of its mycotoxin DON decrease substantially in the presence of soil fauna (earthworms: Lumbricus terrestris, collembolans: Folsomia candida and nematodes: Aphelenchoides saprophilus) in a commercial cropping system managed with conservation tillage located in Northern Germany. The results of our investigations pointed out that the degradation performance of the introduced soil fauna must be considered as an important contribution to the biodegradation of fungal plant diseases and fungal-related contaminants. Different size classes within functional groups and the traits of keystone species appear to be significant for soil function and the provision of ecosystem services as in particular L. terrestris revealed to be the driver of the degradation process. Thus, earthworms contribute to a sustainable control of fungal pathogens like Fusarium and its mycotoxins in wheat straw by reducing the risk of plant diseases and environmental pollution as ecosystem services. Further studies are planned within the EU-project SoilMan under the BiodivERsA network. In context of the suppression of fungal plant pathogens and the detoxification of their mycotoxins by soil organisms in agroecosystems it is hypothesised that (1) processes related to services or disservices are induced and directed by abundance and activity of functional groups of soil biota; (2) dynamics and interaction in the soil biota community control ecosystem function and services.
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Peng, Kai; Jin, Long; Niu, Yan D; Huang, Qianqian; McAllister, Tim A; Yang, Hee Eun; Denise, Hubert; Xu, Zhongjun; Acharya, Surya; Wang, Shunxi; Wang, Yuxi
2017-12-15
Purple prairie clover (PPC; Dalea purpurea Vent.) containing 84.5 g/kg DM of condensed tannin (CT) was ensiled without (Control) or with polyethylene glycol (PEG) for 76 days, followed by 14 days of aerobic exposure. Changes in fermentation characteristics were determined and bacterial and fungal communities were assessed using metagenomic sequencing. Addition of PEG that deactivated CT at ensiling increased ( P < 0.05∼0.001) soluble N, non-protein N, lactic acid, total volatile fatty acids, ammonia N, deoxynivalenol (DON) and ochratoxin A (OTA), but decreased ( P < 0.001) pH and water soluble carbohydrates. Concentration of DON and OTA increased ( P < 0.001) for both silages with the extent of increase being greater for Control than for PEG treated silage during aerobic exposure. The PEG treated silage exhibited higher ( P < 0.01∼0.001) copy numbers of total bacteria, Lactobacillus , yeasts and fungi than Control. Addition of PEG decreased ( P < 0.01) bacterial diversity during both ensiling and aerobic exposure, whereas it increased ( P < 0.05) fungal diversity during aerobic exposure. Addition of PEG at ensiling increased ( P < 0.05) abundances of Lactobacillus and Pediococcus , but decreased ( P < 0.01) abundances of Lactococcus and Leuconostoc Filamentous fungi were found in the microbiome at ensiling and after aerobic exposure, whereas the Bacillus were the dominate bacteria after aerobic exposure. In conclusion, CT decreased protein degradation and improved aerobic stability of silage. These desirable outcomes likely reflect the ability of PPC CT to inhibit those microorganisms involved in lowering silage quality and in the production of mycotoxins. IMPORTANCE The present study reports the effects of condensed tannins on the complex microbial communities involved in ensiling and aerobic exposure of purple prairie clover. This study documents the ability of condensed tannins to lower mycotoxin production and associated microbiome. Taxonomic bacterial community profiles were dominated by the Lactobacillales after fermentation, with a notable increase in Bacillus as a result of aerobic exposure. It is interesting to observe that condensed tannins decreased bacterial diversity during both ensiling and aerobic exposure but increased fungal diversity during aerobic exposure only. The present study indicates that the effects of condensed tannins on microbial communities lead to a reduced lactic acid and total volatile fatty acids production, proteolysis and mycotoxin concentration in the terminal silage and an improved aerobic stability. Condensed tannins could be used as additive to control unfavorable microbial development and maybe enhanced feed safety. © Crown copyright 2017.
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Don Young Don Young Professional IV-ESH Don.Young@nrel.gov | 303-384-7144 Don Young is an EHS Professional at the NWTC. He received his B.S. from Colorado State University in Biological Science before
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Exploring Fusarium head blight disease control by RNA interference
USDA-ARS?s Scientific Manuscript database
RNA interference (RNAi) technology provides a novel tool to study gene function and plant protection strategies. Fusarium graminearum is the causal agent of Fusarium head blight (FHB), which reduces crop yield and quality by producing trichothecene mycotoxins including 3-acetyl deoxynivalenol (3-ADO...
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Portable Infrared Laser Spectroscopy for On-site Mycotoxin Analysis.
Sieger, Markus; Kos, Gregor; Sulyok, Michael; Godejohann, Matthias; Krska, Rudolf; Mizaikoff, Boris
2017-03-09
Mycotoxins are toxic secondary metabolites of fungi that spoil food, and severely impact human health (e.g., causing cancer). Therefore, the rapid determination of mycotoxin contamination including deoxynivalenol and aflatoxin B 1 in food and feed samples is of prime interest for commodity importers and processors. While chromatography-based techniques are well established in laboratory environments, only very few (i.e., mostly immunochemical) techniques exist enabling direct on-site analysis for traders and manufacturers. In this study, we present MYCOSPEC - an innovative approach for spectroscopic mycotoxin contamination analysis at EU regulatory limits for the first time utilizing mid-infrared tunable quantum cascade laser (QCL) spectroscopy. This analysis technique facilitates on-site mycotoxin analysis by combining QCL technology with GaAs/AlGaAs thin-film waveguides. Multivariate data mining strategies (i.e., principal component analysis) enabled the classification of deoxynivalenol-contaminated maize and wheat samples, and of aflatoxin B 1 affected peanuts at EU regulatory limits of 1250 μg kg -1 and 8 μg kg -1 , respectively.
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Portable Infrared Laser Spectroscopy for On-site Mycotoxin Analysis
Sieger, Markus; Kos, Gregor; Sulyok, Michael; Godejohann, Matthias; Krska, Rudolf; Mizaikoff, Boris
2017-01-01
Mycotoxins are toxic secondary metabolites of fungi that spoil food, and severely impact human health (e.g., causing cancer). Therefore, the rapid determination of mycotoxin contamination including deoxynivalenol and aflatoxin B1 in food and feed samples is of prime interest for commodity importers and processors. While chromatography-based techniques are well established in laboratory environments, only very few (i.e., mostly immunochemical) techniques exist enabling direct on-site analysis for traders and manufacturers. In this study, we present MYCOSPEC - an innovative approach for spectroscopic mycotoxin contamination analysis at EU regulatory limits for the first time utilizing mid-infrared tunable quantum cascade laser (QCL) spectroscopy. This analysis technique facilitates on-site mycotoxin analysis by combining QCL technology with GaAs/AlGaAs thin-film waveguides. Multivariate data mining strategies (i.e., principal component analysis) enabled the classification of deoxynivalenol-contaminated maize and wheat samples, and of aflatoxin B1 affected peanuts at EU regulatory limits of 1250 μg kg−1 and 8 μg kg−1, respectively. PMID:28276454
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Mayer, Stefan; Twarużek, Magdalena; Błajet-Kosicka, Anna; Grajewski, Jan
2016-03-01
Manual sorting of onions is known to be associated with a bioaerosol exposure. The study aimed to gain an initial indication as to what extent manual sorting of onions is also associated with mycotoxin exposure. Twelve representative samples of outer onion skins from different onion origins were sampled and analyzed with a multimycotoxin method comprising 40 mycotoxins using a single extraction step followed by liquid chromatography with electrospray ionization and triple quadrupole mass spectrometry. Six of the 12 samples were positive for mycotoxins. In those samples, deoxynivalenol, fumonisin B1, and B2 were observed in quantitatively detectable amounts of 3940 ng/g for fumonisin B1 and in the range of 126-587 ng/g for deoxynivalenol and 55-554 ng/g for fumonisin B2. Although the results point to a lower risk due to mycotoxins, the risk should not be completely neglected and has to be considered in the risk assessment.
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Goossens, Joline; Vandenbroucke, Virginie; Pasmans, Frank; De Baere, Siegrid; Devreese, Mathias; Osselaere, Ann; Verbrugghe, Elin; Haesebrouck, Freddy; De Saeger, Sarah; Eeckhout, Mia; Audenaert, Kris; Haesaert, Geert; De Backer, Patrick; Croubels, Siska
2012-01-01
It is recognized that mycotoxins can cause a variety of adverse health effects in animals, including altered gastrointestinal barrier function. It is the aim of the present study to determine whether mycotoxin-contaminated diets can alter the oral bioavailability of the antibiotics doxycycline and paromomycin in pigs, and whether a mycotoxin adsorbing agent included into diets interacts with those antibiotics. Experiments were conducted with pigs utilizing diets that contained blank feed, mycotoxin-contaminated feed (T-2 toxin or deoxynivalenol), mycotoxin-contaminated feed supplemented with a glucomannan mycotoxin binder, or blank feed supplemented with mycotoxin binder. Diets with T-2 toxin and binder or deoxynivalenol and binder induced increased plasma concentrations of doxycycline administered as single bolus in pigs compared to diets containing blank feed. These results suggest that complex interactions may occur between mycotoxins, mycotoxin binders, and antibiotics which could alter antibiotic bioavailability. This could have consequences for animal toxicity, withdrawal time for oral antibiotics, or public health. PMID:22606377
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Portable Infrared Laser Spectroscopy for On-site Mycotoxin Analysis
NASA Astrophysics Data System (ADS)
Sieger, Markus; Kos, Gregor; Sulyok, Michael; Godejohann, Matthias; Krska, Rudolf; Mizaikoff, Boris
2017-03-01
Mycotoxins are toxic secondary metabolites of fungi that spoil food, and severely impact human health (e.g., causing cancer). Therefore, the rapid determination of mycotoxin contamination including deoxynivalenol and aflatoxin B1 in food and feed samples is of prime interest for commodity importers and processors. While chromatography-based techniques are well established in laboratory environments, only very few (i.e., mostly immunochemical) techniques exist enabling direct on-site analysis for traders and manufacturers. In this study, we present MYCOSPEC - an innovative approach for spectroscopic mycotoxin contamination analysis at EU regulatory limits for the first time utilizing mid-infrared tunable quantum cascade laser (QCL) spectroscopy. This analysis technique facilitates on-site mycotoxin analysis by combining QCL technology with GaAs/AlGaAs thin-film waveguides. Multivariate data mining strategies (i.e., principal component analysis) enabled the classification of deoxynivalenol-contaminated maize and wheat samples, and of aflatoxin B1 affected peanuts at EU regulatory limits of 1250 μg kg-1 and 8 μg kg-1, respectively.
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da Luz, Suzane Rickes; Pazdiora, Paulo Cesar; Dallagnol, Leandro José; Dors, Giniani Carla; Chaves, Fábio Clasen
2017-04-01
Wheat (Triticum aestivum) is an annual crop, cultivated in the winter and spring and susceptible to several pathogens, especially fungi, which are managed with fungicides. It is also one of the most consumed cereals, and can be contaminated by mycotoxins and fungicides. The objective of this study was to validate an analytical method by LC-MS for simultaneous determination of mycotoxins and fungicide residues in wheat grains susceptible to fusarium head blight treated with fungicides, and to evaluate the relationship between fungicide application and mycotoxin production. All parameters of the validated analytical method were within AOAC and ANVISA limits. Deoxynivalenol was the prevalent mycotoxin in wheat grain and epoxiconazole was the fungicide residue found in the highest concentration. All fungicidal treatments induced an increase in AFB2 production when compared to the control (without application). AFB1 and deoxynivalenol, on the contrary, were reduced in all fungicide treatments compared to the control. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Liu, Da-wei; Liu, Hong-yi; Zhang, Hai-bin; Cao, Ming-chang; Sun, Yong; Wu, Wen-da; Lu, Chang-hu
2016-01-01
A survey was conducted to determine whether mycotoxins were present in the foods consumed by red-crowned cranes (Grus japonensis) in the Yancheng Biosphere Reserve, China. Collected in the reserve’s core, buffer, and experimental zones during overwintering periods of 2013 to 2015, a total of 113 food samples were analyzed for aflatoxin B1, deoxynivalenol, zearalenone, T-2 toxin, and ochratoxin A using high performance liquid chromatography (HPLC). The contamination incidences vary among different zones and the mycotoxins levels of different food samples also presented disparity. Average mycotoxin concentration from rice grain was greater than that from other food types. Among mycotoxin-positive samples, 59.3% were simultaneously contaminated with more than one toxin. This study demonstrated for the first time that red-crowned cranes were exposed to mycotoxins in the Yancheng Biosphere Reserve and suggested that artificial wetlands could not be considered good habitats for the birds in this reserve, especially rice fields. PMID:26834016
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Reading in the Social Studies: What Not to Do
ERIC Educational Resources Information Center
Parsons, James; Tomas, Douglas
1978-01-01
Five points are: (1) don't assume students know how to use their textbooks; (2) don't ignore the problem or take only short term measures for improvement; (3) don't give undirected assignments; (4) don't force students to pronounce every word correctly; (5) don't send students to the dictionary for words they don't know. (Author/JK)
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USDA-ARS?s Scientific Manuscript database
Dissolved organic nitrogen (DON) transport from animal agriculture to surface waters can lead to eutrophication and dissolved oxygen depletion. Biodegradable DON (BDON) is a portion of DON that is mineralized by bacteria while bioavailable DON (ABDON) is utilized by bacteria and/or algae. This stu...
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USDA-ARS?s Scientific Manuscript database
Fusarium Head Blight is a disease of cereal crops that causes severe yield losses and mycotoxin contamination of grain. The main causal pathogen, Fusarium graminearum, produces the trichothecene toxins deoxynivalenol or nivalenol as virulence factors. Nivalenol-producing isolates are most prevalent ...
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Genetic and phenotypic diversity within the Fusarium graminearum species complex in Norway
USDA-ARS?s Scientific Manuscript database
As has been observed in several European countries, the frequency of Fusarium head blight (FHB) caused by members of the Fusarium graminearum species complex (FGSC) has increased in Norwegian cereals in recent years, resulting in elevated levels of deoxynivalenol in cereal grains. The objective of t...
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USDA-ARS?s Scientific Manuscript database
A comprehensive quantum chemical study was carried out on 34 type A and type B trichothecenes, including selected derivatives and biosynthetic precursors of deoxynivalenol, nivalenol, and T-2 toxin. Quantum parameters, Natural Bond Orbital (NBO) analysis, and molecular properties were calculated on ...
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USDA-ARS?s Scientific Manuscript database
Filamentous fungi that contaminate livestock feeds and human food supply often produce toxigenic secondary metabolites known as mycotoxins. Among the hundreds of known mycotoxins, aflatoxins, deoxynivalenol, fumonisins, ochratoxin A and zearalenone are considered the most commercially important. In...
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Spacsuit donning and doffing in zero-g training for Don Peterson STS-6
NASA Technical Reports Server (NTRS)
1982-01-01
Spacsuit donning and doffing in zero-g training for Don Peterson of the STS-6 crew. The training is being held aboard the KC-135 to simulate weightlessness. He is being assisted to don the lower torso of the extravehicular mobility unit (EMU) by an ILC technician.
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McKinney, Selina H; Corazzini, Kirsten; Anderson, Ruth A; Sloane, Richard; Castle, Nicholas G
2016-01-01
Nursing homes are becoming increasingly complex clinical environments because of rising resident acuity and expansion of postacute services within a context of historically poor quality performance. Discrete quality markers have been linked to director of nursing (DON) leadership behaviors. However, the impact of DON leadership across all measured areas of DON jurisdiction has not been tested using comprehensive domains of quality deficiencies. The aim of this study was to examine the effects of DON leadership style including behaviors that facilitate the exchange of information between diverse people on care quality domains through the lens of complexity science. Three thousand six hundred nine DONs completed leadership and intent-to-quit surveys. Quality markers that were deemed DON sensitive included all facility survey deficiencies in the domains of resident behaviors/facility practices, quality of life, nursing services, and quality of care. Logistic regression procedures estimated associations between variables. The odds of deficiencies for all DON sensitive survey domains were lower in facilities where DONs practiced complexity leadership including more staff input and shared decisional authority. DON quit intentions were aligned with higher odds of facility deficiencies across all domains. Results supported the hypotheses that DONs using complexity leadership approaches by interacting more freely with staff, discussing resident issues, and sharing decision making produced better care outcomes from every DON sensitive metric assessed by Centers for Medicare and Medicaid Services. The mechanism linking poor quality with high DON quit intentions is an area for future research. Encouraging DON use of complexity leadership approaches has the potential to improve a broad swath of quality outcomes.
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Simsek, Halis; Kasi, Murthy; Ohm, Jae-Bom; Murthy, Sudhir; Khan, Eakalak
2016-04-01
Dissolved organic nitrogen (DON) and its biodegradability in treated wastewater have recently gained attention due to increased regulatory requirements on effluent quality to protect receiving waters. Laboratory scale chemostat experiments were conducted at 9 different solids retention times (SRTs) (0.3, 0.7, 2, 3, 4, 5, 7, 8, and 13 days) to examine whether SRT could be used to control DON, biodegradable DON (BDON), and DON biodegradability (BDON/DON) levels in treated wastewater. Results indicated no trend between effluent DON and SRTs. Effluent BDON was comparable for SRTs of 0.3-4 days and had a decreasing trend with SRT after that. Effluent DON biodegradability (effluent BDON/effluent DON) ranging from 23% to 59% tended to decrease with SRT. Chemostat during longer SRTs, however, was contributing to non-biodegradable DON (NBDON) and this fraction of DON increased with SRT above 4 days. Model calibration results indicated that ammonification rate, and growth rates for ordinary heterotrophs, ammonia oxidizing bacteria and nitrite oxidizing bacteria were not constants but have a decreasing trend with increasing SRT. This study indicates the benefit of high SRTs in term of producing effluent with less DON biodegradability leading to relatively less oxygen consumption and nutrient support in receiving waters. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Assuming the mantle of leadership: issues and challenges for directors of nursing.
Fleming, Mary Louise; Kayser-Jones, Jeanie
2008-11-01
This ethnographic study investigated leadership from the perspective of directors of nursing (DONs) in proprietary nursing homes. Data from interviews and extensive participant observation with 10 DONs were analyzed using open coding and content analysis. The study drew on a priori concepts from transformational leadership theory to describe the role, DON approaches to leadership, and factors that facilitate or impede leadership in nursing homes. This article reports findings from a larger study related to conditions existing when participants entered the DON position. Antecedent conditions influenced organizational expectations of incoming DONs and shaped participants' leadership experiences. DONs filling long-standing vacancies had to reestablish the influence and authority of the role. Those replacing unsuccessful DONs confronted serious regulatory, care, and morale issues. In contrast, DONs with successful predecessors experienced organizational support and had confidence in their abilities to lead.
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Sun, Jingyi; Khan, Eakalak; Simsek, Senay; Ohm, Jae-Bom; Simsek, Halis
2017-11-01
Dissolved organic nitrogen (DON) from animal wastes can contribute to pollution of surface waters. Bioavailable DON (ABDON) is a portion of DON utilized by algae with or without bacteria. This study determined DON and ABDON levels in animal wastewater collected from two different sources: an animal feedlot wastewater storage tank and a sheep wastewater storage lagoon. Inocula for the ABDON bioassays were comprised of individual species and several combinations involving two algae (Chlamydomonas reinhardtii and Chlorella vulgaris) and a mixed liquor suspended solids (MLSS) bacterial culture. The ratio of initial DON to initial total dissolved nitrogen was 18% in the feedlot wastewater samples and 70% in the lagoon wastewater samples. The results showed that between 1.6 and 4.5 mg-NL-1 DON (45-79% of initial DON) in the feedlot samples and between 3.4 and 7.5 mg-NL-1 DON (36%-79% of initial DON) in the lagoon samples were bioavailable with the inocula tested. These results suggest that when considering eutrophication potential of livestock wastewater, organic nitrogen should be included in addition to the obvious culprits, ammonia and nitrate. Copyright © 2017 Elsevier Ltd. All rights reserved.
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USDA-ARS?s Scientific Manuscript database
Fusarium head blight (FHB) or scab, caused by Fusarium graminearum Schwabe, can cause significant economic losses in small grain production. There is a need to develop effective management strategies for FHB. Five field experiments were conducted from 2007 to 2009 to determine the effects of integ...
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1988-07-25
laboratory studies, but not useful for evaluation of natural exposure. Several conventional immunoassay techniques based on tritiated ligands (Fontelo. et...1978. Natural occurrence of trichothecenes (nivaienol, deoxynivalenol, T-2) and zearalenone in corn. ExperiLentia 34, 1333-1334. LEE. S. and CHU, F.S
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Operational Environmental Assessment
1988-09-01
Chemistry Branch - Physical Chemistry Branch " Analytical Research Division - Analytical Systems Branch - Methodology Research Branch - Spectroscopy Branch...electromagnetic frequency spec- trum and includes radio frequencies, infrared , visible light, ultraviolet, X-rays and gamma rays (in ascending order of...Verruculogen Aflatrem Picrotoxin Ciguatoxin Mycotoxins Simple Tr ichothecenes T-2 Toxin T-2 Tetraol Neosolaniol * Nivalenol Deoxynivalenol Verrucarol B-3 B lank
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USDA-ARS?s Scientific Manuscript database
The TrigoCor strain of Bacillus amyloliquefaciens provides consistent control against Fusarium head blight of wheat in controlled settings but there is a lack of disease and deoxynivalenol suppression in field settings. Since production of antifungal compounds is thought to be the main mode of actio...
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Miró-Abella, Eugènia; Herrero, Pol; Canela, Núria; Arola, Lluís; Borrull, Francesc; Ras, Rosa; Fontanals, Núria
2017-08-15
A method was developed for the simultaneous determination of 11 mycotoxins in plant-based beverage matrices, using a QuEChERS extraction followed by ultra-high performance liquid chromatography coupled to tandem mass spectrometry detection (UHPLC-(ESI)MS/MS). This multi-mycotoxin method was applied to analyse plant-based beverages such as soy, oat and rice. QuEChERS extraction was applied obtaining suitable extraction recoveries between 80 and 91%, and good repeatability and reproducibility values. Method Quantification Limits were between 0.05μgL -1 (for aflatoxin G 1 and aflatoxin B 1 ) and 15μgL -1 (for deoxynivalenol and fumonisin B 2 ). This is the first time that plant-based beverages have been analysed, and certain mycotoxins, such as deoxynivalenol, aflatoxin B 1 , aflatoxin B 2 , aflatoxin G 1 , aflatoxin G 2 , ochratoxin A, T-2 toxin and zearalenone, were found in the analysed samples, and some of them quantified between 0.1μgL -1 and 19μgL -1 . Copyright © 2017 Elsevier Ltd. All rights reserved.
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Hamed, Ahmed M; Arroyo-Manzanares, Natalia; García-Campaña, Ana M; Gámiz-Gracia, Laura
2017-11-01
Vegetable milks are considered as functional foods due to their physiological benefits. Although the consumption of these products has significantly increased, they have received little attention in legislation with regard to contaminants. However, they may contain mycotoxins resulting from the use of contaminated raw materials. In this work, ultra-high-performance liquid chromatography tandem mass spectrometry has been proposed for the determination of the most relevant Fusarium toxins (fumonisin B 1 and B 2 , HT-2 and T-2 toxins, zearalenone, deoxynivalenol and fusarenon-X) in different functional beverages based on cereals, legumes and seeds. Sample treatment consisted of a simple salting-out-assisted liquid-liquid extraction with no further clean-up. The method provided limits of quantification between 3.2 and 57.7 µg L -1 , recoveries above 80% and precision with RSD lower than 12%. The method was also applied for studying the occurrence of these mycotoxins in market samples of vegetable functional beverages and deoxynivalenol was found in three oat-based commercial drinks.
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Mycotoxin production and cytotoxicity of Fusarium strains isolated from Norwegian cereals.
Langseth, W; Bernhoft, A; Rundberget, T; Kosiak, B; Gareis, M
Thirty-four isolates of the eight most common Fusarium species isolated from Norwegian cereals; F. avenaceum, F. culmorum, F. equiseti, F. graminearum, F. poae, F. sporotrichioides, F. torulosum and F. tricinctum were studied for their cytotoxicity and ability to produce mycotoxins. The strains were cultivated on rice, and analysed for trichothecenes (all species), zearalenone (all species), fusarochromanone (F. equiseti), wortmannin (F. torulosum), moniliformin and enniatins (F. avenaceum, F. tricinctum and F. torulosum). The cytotoxicity of the extracts were examined with an (in vitro) MTT-cell culture assay. All F. graminearum and five of seven F. culmorum isolates belonged to chemotype IA, producing deoxynivalenol and 3-acetyl-deoxynivalenol, while the two other F. culmorum strains were nivalenol producers (chemotype II). The F. equiseti isolates and one of the F. poae isolates produced both type A and B trichothecenes, and relatively large quantities of fusarochromanone were detected in the F. equiseti cultures. All Fusarium species studied showed significant cytotoxicity, but with a large variation between species, and also within each species. F. sporotrichioides and F. equiseti showed the highest average cytotoxicity.
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Wu, Li; Wang, Bujun
2015-10-15
The present study investigated the changes and conversion profiles of DON, its conjugations 3-ADON, and 15-ADON during bread making process, by spiking targeted mycotoxin standards to Fusarium mycotoxins-free wheat flour. No significant (p < 0.05) changes of DON levels were observed during dough preparation stages, including kneading, fermentation, and proofing. A reduction of DON level ranged from 4% to 14% was observed during baking process. The main thermal degradation products of DON, namely norDON A, B, C, and F were detected in the bread crust. Regarding ADONs, decreases of 20-40% for 3-ADON and 28-60% for 15-ADON were found during fermentation stage, and further losses of ADONs were observed after proofing process. Although ADONs levels gained an increase after baking. This study demonstrated that ADONs were converted to DON, while no ADONs were detectable in DON spiked samples during bread making process. The mechanism that ADONs could be converted into DON is unclear so far. Copyright © 2015. Published by Elsevier Ltd.
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Lim, Chee Wei; Tai, Siew Hoon; Lee, Lin Min; Chan, Sheot Harn
2012-07-01
The current food crisis demands unambiguous determination of mycotoxin contamination in staple foods to achieve safer food for consumption. This paper describes the first accurate LC-MS/MS method developed to analyze tricothecenes in grains by applying multiple reaction monitoring (MRM) transition and MS(3) quantitation strategies in tandem. The tricothecenes are nivalenol, deoxynivalenol, deoxynivalenol-3-glucoside, fusarenon X, 3-acetyl-deoxynivalenol, 15-acetyldeoxynivalenol, diacetoxyscirpenol, and HT-2 and T-2 toxins. Acetic acid and ammonium acetate were used to convert the analytes into their respective acetate adducts and ammonium adducts under negative and positive MS polarity conditions, respectively. The mycotoxins were separated by reversed-phase LC in a 13.5-min run, ionized using electrospray ionization, and detected by tandem mass spectrometry. Analyte-specific mass-to-charge (m/z) ratios were used to perform quantitation under MRM transition and MS(3) (linear ion trap) modes. Three experiments were made for each quantitation mode and matrix in batches over 6 days for recovery studies. The matrix effect was investigated at concentration levels of 20, 40, 80, 120, 160, and 200 μg kg(-1) (n = 3) in 5 g corn flour and rice flour. Extraction with acetonitrile provided a good overall recovery range of 90-108% (n = 3) at three levels of spiking concentration of 40, 80, and 120 μg kg(-1). A quantitation limit of 2-6 μg kg(-1) was achieved by applying an MRM transition quantitation strategy. Under MS(3) mode, a quantitation limit of 4-10 μg kg(-1) was achieved. Relative standard deviations of 2-10% and 2-11% were reported for MRM transition and MS(3) quantitation, respectively. The successful utilization of MS(3) enabled accurate analyte fragmentation pattern matching and its quantitation, leading to the development of analytical methods in fields that demand both analyte specificity and fragmentation fingerprint-matching capabilities that are unavailable under MRM transition.
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Urusov, Alexandr E; Gubaidullina, Miliausha K; Petrakova, Alina V; Zherdev, Anatoly V; Dzantiev, Boris B
2017-12-06
A new kind of competitive immunochromatographic assay is presented. It is based on the use of a test strip loaded with (a) labeled specific antibodies, (b) a hapten-protein conjugate at the control zone, and (c) antibodies interacting with the specific antibodies in the analytical zone. In the case where a sample does not contain the target antigen (hapten), all labeled antibodies remain in the control zone because of the selected ratio of reactants. The analytical zone remains colorless because the labeled antibodies do not reach it. If an antigen is present in the sample, it interferes with the binding of the specific antibodies in the control zone and knocks them out. Some of these antibodies pass the control zone to form a colored line in the analytical zone. The intensity of the color is directly proportional to the amount of the target antigen in the sample. The assay has an attractive feature in that an appearance in coloration is more easily detected visually than a decoloration. Moreover, the onset of coloration is detectable at a lower concentration than a decoloration. The new detection scheme was applied to the determination of the mycotoxin deoxynivalenol. The visual limit of detection is 2 ng·mL -1 in corn extracts (35 ng per gram of sample). With the same reagents, this is lower by a factor of 60 than the established test strip. The assay takes only 15 min. This new kind of assay has wide potential applications for numerous low molecular weight analytes. Graphical abstract Competitive immunochromatography with direct analyte-signal dependence is proposed. It provides a 60-fold decrease of the detection limit for mycotoxin deoxynivalenol. The analyte-antibody-label complexes move along the immobilized antigen (control zone) and bind with anti-species antibodies (test zone).
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Dissolved organic nitrogen in urban streams: Biodegradability and molecular composition studies.
Lusk, Mary G; Toor, Gurpal S
2016-06-01
A portion of the dissolved organic nitrogen (DON) is biodegradable in water bodies, yet our knowledge of the molecular composition and controls on biological reactivity of DON is limited. Our objective was to investigate the biodegradability and molecular composition of DON in streams that drain a gradient of 19-83% urban land use. Weekly sampling over 21 weeks suggested no significant relationship between urban land use and DON concentration. We then selected two streams that drain 28% and 83% urban land use to determine the biodegradability and molecular composition of the DON by coupling 5-day bioassay experiments with high resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Both urban streams contained a wide range of N-bearing biomolecular formulas and had >80% DON in lignin-like compounds, with only 5-7% labile DON. The labile DON consisted mostly of lipid-and protein-like structures with high H/C and low O/C values. Comparison of reactive formulas and formed counterparts during the bioassay experiments indicated a shift toward more oxygenated and less saturated N-bearing DON formulas due to the microbial degradation. Although there was a little net removal (5-7%) of organic-bound N over the 5-day bioassay, there was some change to the carbon skeleton of DON compounds. These results suggest that DON in urban streams contains a complex mixture of compounds such as lipids, proteins, and lignins of variable chemical structures and biodegradability. Copyright © 2016 Elsevier Ltd. All rights reserved.
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van der Fels-Klerx, H J; Olesen, J E; Naustvoll, L-J; Friocourt, Y; Mengelers, M J B; Christensen, J H
2012-01-01
Climate change is expected to affect food and feed safety, including the occurrence of natural toxins in primary crop and seafood production; however, to date, quantitative estimates are scarce. This study aimed to estimate the impact of climate change effects on mycotoxin contamination of cereal grains cultivated in the terrestrial area of north west Europe, and on the frequency of harmful algal blooms and contamination of shellfish with marine biotoxins in the North Sea coastal zone. The study focused on contamination of wheat with deoxynivalenol, and on abundance of Dinophysis spp. and the possible relationship with diarrhetic shellfish toxins. The study used currently available data and models. Global and regional climate models were combined with models of crop phenology, mycotoxin prediction models, hydrodynamic models and ecological models, with the output of one model being used as input for the other. In addition, statistical data analyses using existing national datasets from the study area were performed to obtain information on the relationships between Dinophysis spp. cell counts and contamination of shellfish with diarrhetic shellfish toxins as well as on frequency of cereal cropping. In this paper, a summary of the study is presented, and overall conclusions and recommendations are given. Climate change projections for the years 2031-2050 were used as the starting point of the analyses relative to a preceding 20-year baseline period from which the climate change signal was calculated. Results showed that, in general, climate change effects lead to advanced flowering and harvest of wheat, and increased risk of contamination of wheat with deoxynivalenol. Blooms of dinoflagellates were estimated to occur more often. If the group of Dinophysis spp. behaves similarly to other flagellates in the future then frequency of harmful algal blooms of Dinophysis spp. may also increase, but consequences for contamination of shellfish with diarrhetic shellfish toxins are uncertain. Climate change will also have indirect effects on toxin contamination, which may be equally important. For example, the frequency of cropping of wheat and maize in north Europe was projected to increase under climate change, which will also increase the risk of contamination of the grains with deoxynivalenol. Risk managers are encouraged to consider the entire range of the predictions of climate change effects on food safety hazards, rather than median or average values only. Furthermore, it is recommended to closely monitor levels of mycotoxins and marine biotoxins in the future, in particular related to risky situations associated with favourable climatic conditions for toxin producing organisms. In particular, it is important to pay attention to the continuity of collecting the right data, and the availability and accessibility of databases. On a European level, it is important to stress the need for harmonisation of terminology and data collection.
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[Removal of DON in micro-polluted raw water by coagulation and adsorption using activated carbon].
Liu, Bing; Yu, Guo-Zhong; Gu, Li; Zhao, Cheng-Mei; Li, Qing-Fei; Zhai, Hui-Min
2013-04-01
Dissolved organic nitrogen as a precursor of new type nitrogenous disinfection by-products in drinking water attracted gradually the attention of scholars all over the world. In order to explore the mechanism of DON removal in micro-polluted raw water by coagulation and adsorption, water quality parameters, such as DON, DOC, NH4(+) -N, UV254, pH and dissolved oxygen, were determined in raw water and the molecular weight distribution of the DON and DOC was investigated. The variations in DON, DOC and UV254 in the coagulation and adsorption tests were investigated, and the changes of DON in raw water were characterized using three-dimensional fluorescence spectroscopy. The results showed that DON, DOC and UV254 were 1.28 mg x L(-1), 8.56 mg x L(-1), 0.16 cm(-1), and DOC/DON and SUVA were 6.69 mg x mg(-1), 1.87 m(-1) x (mg x L(-1))(-1) in raw water, respectively. The molecular weight distribution of the DON in raw water showed a bimodal distribution. The small molecular weight (< 6 000) fractions accounted for a high proportion of 68% and the large (> 20 000) fractions accounted for about 22%. The removal of DON, DOC and UV254 was about 20%, 26% and 70%, respectively, in the coagulation test and the dosage of coagulant was 10 mg x L(-1). The removal of DON, DOC and UV254 was about 60%, 35% and 100%, respectively, in the adsorption test and the dosage of activated carbon was 1.0 g. In the combination of coagulation and adsorption, the removal of DON and DOC reached approximately 82% and 64%, respectively. 3DEEM revealed that the variation of DON in the coagulation and adsorption tests depended intimately on tryptophan protein-like substances, aromatic protein-like substances and fulvic acid-like substances.
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Li, Keqiang; Ma, Yunpeng; Dai, Aiquan; Wang, Xiulin
2017-11-30
Dissolved organic nitrogen (DON) is the major nitrogen form in the Bohai Sea. Land-based DON is released into the nitrogen pool and degraded by planktonic microbiota in coastal ocean. In this study, we evaluated the degradation of land-based DON, particularly its dynamics and bioavailability, in coastal water by linking experiment and modeling. Results showed that the degradation rate constant of DON from sewage treatment plant was significantly faster than those of other land-based sources (P<0.05). DON was classified into three categories based on dynamics and bioavailability. The supply of dissolved inorganic nitrogen (DIN) pool from the DON pool of Liao River, Hai River, and Yellow River was explored using a 3D hydrodynamic multi-DON biogeochemical model in the Bohai Sea. In the model, large amounts of DIN were supplied from DON of Liao River than the other rivers because of prolonged flushing time in Liaodong Bay. Copyright © 2017 Elsevier Ltd. All rights reserved.
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SPACESUIT DONNING AND DOFFING - ZERO-G TRAINING - DON PETERSON - STS-6
1982-07-14
Spacesuit Donning and Doffing in Zero-G Training for Don Peterson of the STS-6 Crew with Astronaut Jerry Ross assisting; and, apparatus for testing the JSC Mechanically-Induced Settling Technology (MIST) Experiment. The training is being held aboard the KC-135 to simulate weightlessness. He is being assisted to don the lower torso of the Extravehicular Mobility Unit (EMU) by an ILC Technician. 1. ASTRONAUT ROSS, JERRY L. - ZERO-G SUITING 2. SHUTTLE - EXPERIMENTS (MIST)
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Evaluation of leachate dissolved organic nitrogen discharge effect on wastewater effluent quality.
Bolyard, Stephanie C; Reinhart, Debra R
2017-07-01
Nitrogen is limited more and more frequently in wastewater treatment plant (WWTP) effluents because of the concern of causing eutrophication in discharge waters. Twelve leachates from eight landfills in Florida and California were characterized for total nitrogen (TN) and dissolved organic nitrogen (DON). The average concentration of TN and DON in leachate was approximately 1146mg/L and 40mg/L, respectively. Solid-phase extraction was used to fractionate the DON based on hydrophobic (recalcitrant fraction) and hydrophilic (bioavailable fraction) chemical properties. The average leachate concentrations of bioavailable (bDON) and recalcitrant (rDON) DON were 16.5mg/L and 18.4mg/L, respectively. The rDON fraction was positively correlated, but with a low R 2 , with total leachate apparent color dissolved UV 254 , chemical oxygen demand (COD), and humic acid (R 2 equals 0.38, 0.49, and 0.40, respectively). The hydrophobic fraction of DON (rDON) was highly colored. This fraction was also associated with over 60% of the total leachate COD. Multiple leachate and wastewater co-treatment simulations were carried out to assess the effects of leachate on total nitrogen wastewater effluent quality using removals for four WWTPs under different scenarios. The calculated pass through of DON suggests that leachate could contribute to significant amounts of nitrogen discharged to aquatic systems. Copyright © 2017 Elsevier Ltd. All rights reserved.
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NASA Astrophysics Data System (ADS)
Chen, You-Xin; Chen, Hung-Yu; Wang, Wei; Yeh, Jun-Xian; Chou, Wen-Chen; Gong, Gwo-Ching; Tsai, Fu-Jung; Huang, Shih-Jen; Lin, Cheng-Ting
2015-07-01
In this study, we collected and analyzed rainwater samples from Keelung, Taiwan, which is a coastal city located south of the East China Sea (ECS). From January 2012 until June 2013, 78 rainwater samples were collected over an 18-month period and were analyzed to examine the total dissolved nitrogen (TDN) and major ions in the rainwater. TDN can be divided into dissolved inorganic nitrogen (DIN) and dissolved organic nitrogen (DON). This study, which focused on the composition of DON, is the first study to employ ultrafiltration to separate DON in wet deposition into low molecular weight-DON (LMW-DON) and high molecular weight-DON (HMW-DON). The concentrations of dissolved nitrogen species observed in the research area between November 2012 and April 2013 were relatively high, whereas those observed between May 2013 and October 2012 were relatively low. The patterns of changes over time were similar to those of non-sea-salt (nss) ions. The concentration of nss-ions was high during months in which the total dissolved nitrogen concentration was also high, which occur frequently during the spring and winter. In addition, the concentration of nss-ions was low during months in which the TDN concentration was low, which primarily occurs during the summer. The amounts of DIN and DON accounted for 63 ± 5% and 37 ± 5% of the TDN, respectively, and the percentage of the DIN was higher during the spring and winter. The concentrations of LMW-DON and HMW-DON, which accounted for 84 ± 3% and 16 ± 3% of the DON, respectively, were both high in the winter and low in the summer. The percentage of LMW-DON increased in the summer, possibly because of the numerous oceanic air masses and typhoons. Furthermore, the percentage of HMW-DON increased in the spring, potentially due to biomass burning during agricultural activities. Regarding the wet deposition fluxes, the DIN (197 ± 10.27 mmol m-2 yr-1) and DON (129 ± 6.82 mmol m-2 yr-1) accounted for approximately 64% and 36% of the TDN, respectively, and the LMW-DON (108 ± 5.97 mmol m-2 yr-1) and HMW-DON (19 ± 1.02 mmol m-2 yr-1) accounted for 85% and 15% of the DON, respectively. The wet deposition flux of the nitrogen species observed in the research area was 332 ± 16.0 mmol m-2 yr-1, and the total flux (dry and wet deposition) was approximately 393.4 ± 25.2 mmol m-2 yr-1 (5508 ± 353 mg N m-2 yr-1).
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Interaction effects of enniatin B, deoxinivalenol and alternariol in Caco-2 cells.
Fernández-Blanco, Celia; Font, Guillermina; Ruiz, Maria-Jose
2016-01-22
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). Copyright © 2015. Published by Elsevier Ireland Ltd.
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USDA-ARS?s Scientific Manuscript database
Fusarium graminearum Schwabe of the ‘3ADON’ chemotype is now displacing ‘15ADON’ isolates in Canada. One concern regarding this shift in chemotypes is related to potential differences in fungicide sensitivity. This could have significant implications as fungicide application is an important strate...
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USDA-ARS?s Scientific Manuscript database
Surveys for crown rot (FCR) and head blight (FHB) of Algerian wheat conducted during 2014 and 2015 revealed that Fusarium culmorum strains producing 3-acetyl-deoxynivalenol (3ADON) or nivalenol (NIV) were the primary causal agents of these important diseases. Morphological identification of the isol...
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USDA-ARS?s Scientific Manuscript database
Mycotoxin contamination in food and feed crops is a major concern worldwide. Fungal pathogens of the genera Aspergillus, Fusarium, and Penicillium are a major threat to food and feed crops due to production of mycotoxins such as aflatoxins, 4-deoxynivalenol, patulin, and numerous other toxic seconda...
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Code of Federal Regulations, 2010 CFR
2010-07-01
... prefers non-destructive, in situ research on DON ship and aircraft wrecks, it recognizes that site... DON ship and aircraft wreck on an individual basis. In some cases, the removal of DON ship and... of DON ship and aircraft wrecks may be justified in specific cases where the existence of a cultural...
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Code of Federal Regulations, 2014 CFR
2014-07-01
... prefers non-destructive, in situ research on DON ship and aircraft wrecks, it recognizes that site... DON ship and aircraft wreck on an individual basis. In some cases, the removal of DON ship and... of DON ship and aircraft wrecks may be justified in specific cases where the existence of a cultural...
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Code of Federal Regulations, 2013 CFR
2013-07-01
... prefers non-destructive, in situ research on DON ship and aircraft wrecks, it recognizes that site... DON ship and aircraft wreck on an individual basis. In some cases, the removal of DON ship and... of DON ship and aircraft wrecks may be justified in specific cases where the existence of a cultural...
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Code of Federal Regulations, 2011 CFR
2011-07-01
... prefers non-destructive, in situ research on DON ship and aircraft wrecks, it recognizes that site... DON ship and aircraft wreck on an individual basis. In some cases, the removal of DON ship and... of DON ship and aircraft wrecks may be justified in specific cases where the existence of a cultural...
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Code of Federal Regulations, 2012 CFR
2012-07-01
... prefers non-destructive, in situ research on DON ship and aircraft wrecks, it recognizes that site... DON ship and aircraft wreck on an individual basis. In some cases, the removal of DON ship and... of DON ship and aircraft wrecks may be justified in specific cases where the existence of a cultural...
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33 CFR 208.82 - Hetch Hetchy, Cherry Valley, and Don Pedro Dams and Reservoirs.
Code of Federal Regulations, 2011 CFR
2011-07-01
... Don Pedro Dams and Reservoirs. 208.82 Section 208.82 Navigation and Navigable Waters CORPS OF..., Cherry Valley, and Don Pedro Dams and Reservoirs. The Turlock Irrigation District and Modesto Irrigation District, acting jointly, hereinafter called the Districts, shall operate Don Pedro Dam and Reservoir in...
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33 CFR 208.82 - Hetch Hetchy, Cherry Valley, and Don Pedro Dams and Reservoirs.
Code of Federal Regulations, 2010 CFR
2010-07-01
... Don Pedro Dams and Reservoirs. 208.82 Section 208.82 Navigation and Navigable Waters CORPS OF..., Cherry Valley, and Don Pedro Dams and Reservoirs. The Turlock Irrigation District and Modesto Irrigation District, acting jointly, hereinafter called the Districts, shall operate Don Pedro Dam and Reservoir in...
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Alt, Jesse; Potter, Michelle C.; Rojas, Camilo; Slusher, Barbara S.
2015-01-01
Glutamine is an abundant amino acid that plays pivotal roles in cell growth, cell metabolism and neurotransmission. Dysregulation of glutamine-utilizing pathways has been associated with pathological conditions such as cancer and neurodegenerative diseases. 6-Diazo-5-Oxo-L-Norleucine (DON) is a reactive glutamine analog that inhibits enzymes affecting glutamine metabolism such as glutaminase, 2-N-amidotransferase, L-asparaginase and several enzymes involved in pyrimidine and purine de novo synthesis. As a result, DON is actively used in preclinical models of cancer and neurodegenerative disease. Moreover, there have been several clinical trials using DON to treat a variety of cancers. Considerations of dose and exposure are especially important with DON treatment due to its narrow therapeutic window and significant side effects. Consequently, a robust quantification bioassay is of interest. DON is a polar unstable molecule which has made quantification challenging. Here we report on the characterization of a bioanalytical method to quantify DON in tissue samples involving DON derivatization with 3N HCl in butanol. The derivatized product is lipophilic and stable. Detection of this analyte by mass spectrometry is fast, specific and can be used to quantify DON in plasma and brain tissue with a limit of detection in the low nanomolar level. PMID:25584882
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Crosby, Heith A; Ihnat, Michael; Miller, Kenneth E
2018-01-01
6-diazo-5-oxo-l-norleucine (DON) is a glutamine antagonist produced naturally by Streptomyces. It inhibits several glutamine-dependent enzyme pathways. Of particular note is its inhibitory effect on the mitochondrial enzyme, glutaminase (GLS), the primary producer of neuronal glutamate. Glutamate is an excitatory neurotransmitter released by primary sensory peripheral nerve terminals and spinal synaptic terminals during pain signaling. Previous work using the tail incision and inflammatory models of pain has demonstrated that a single application of the glutaminase inhibitor, DON, into a surgical incision or the paw of arthritic animals results in pain relief. Even though this compound shows promise as a therapeutic agent, limited data exist regarding its dermal toxicity. As a first approach, we evaluated the effect of several concentrations of DON, on the viability, mitochondrial oxidative capacity and proliferation of rat skin fibroblasts, and then examined the effect of DON after incubation with human liver microsomes on proliferation. Finally, we evaluated DON treated rat skin (tail and hind paw) for cellular necrosis, inflammation and mitotic bodies. No significant effects (p > 0.05) of DON were noted on apoptosis, necrosis, and mitochondrial activity in experiments with cultured rat skin fibroblasts. Flow cytometry revealed the absence of apoptosis in cells treated at the IC50 of 232.5 μM. Enhanced toxicity post-exposure to human microsomes was not observed when compared to DON alone. The H&E staining of the rat skin revealed no obvious pathology in the DON treatment group (10 mM). DON has no/minimal cellular toxicity in vitro on dermal fibroblasts at concentrations that effectively provide analgesia. The local application of concentrations greater than the in vitro IC50 for DON revealed no in vivo skin toxicity. These data provide results indicating zero-to-minimal cellular toxicity with DON and support the further investigation of DON as an analgesic. PMID:29750203
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The Status of Nonagreeing "Don't" and Theories of Root Infinitives
ERIC Educational Resources Information Center
Schutze, Carson T.
2010-01-01
This paper examines two issues concerning nonagreeing "don't" in child English, e.g., "He don't fit". (1) Do children know that "don't" consists of auxiliary "do" plus sentential negation, or do they misanalyze it simply as negation? I argue that the former claim yields both empirical (distributional) and conceptual advantages, while the latter…
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Don Quixote Pond: A Small Scale Model of Weathering and Salt Accumulation
NASA Technical Reports Server (NTRS)
Englert, P.; Bishop, J. L.; Patel, S. N.; Gibson, E. K.; Koeberl, C.
2015-01-01
The formation of Don Quixote Pond in the North Fork of Wright Valley, Antarctica, is a model for unique terrestrial calcium, chlorine, and sulfate weathering, accumulation, and distribution processes. The formation of Don Quixote Pond by simple shallow and deep groundwater contrasts more complex models for Don Juan Pond in the South Fork of Wright Valley. Our study intends to understand the formation of Don Quixote Pond as unique terrestrial processes and as a model for Ca, C1, and S weathering and distribution on Mars.
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Berisha, Arton; Dold, Sebastian; Guenther, Sabine; Desbenoit, Nicolas; Takats, Zoltan; Spengler, Bernhard; Römpp, Andreas
2014-08-30
An ideal method for bioanalytical applications would deliver spatially resolved quantitative information in real time and without sample preparation. In reality these requirements can typically not be met by a single analytical technique. Therefore, we combine different mass spectrometry approaches: chromatographic separation, ambient ionization and imaging techniques, in order to obtain comprehensive information about metabolites in complex biological samples. Samples were analyzed by laser desorption followed by electrospray ionization (LD-ESI) as an ambient ionization technique, by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging for spatial distribution analysis and by high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS) for quantitation and validation of compound identification. All MS data were acquired with high mass resolution and accurate mass (using orbital trapping and ion cyclotron resonance mass spectrometers). Grape berries were analyzed and evaluated in detail, whereas wheat seeds and mouse brain tissue were analyzed in proof-of-concept experiments. In situ measurements by LD-ESI without any sample preparation allowed for fast screening of plant metabolites on the grape surface. MALDI imaging of grape cross sections at 20 µm pixel size revealed the detailed distribution of metabolites which were in accordance with their biological function. HPLC/ESI-MS was used to quantify 13 anthocyanin species as well as to separate and identify isomeric compounds. A total of 41 metabolites (amino acids, carbohydrates, anthocyanins) were identified with all three approaches. Mass accuracy for all MS measurements was better than 2 ppm (root mean square error). The combined approach provides fast screening capabilities, spatial distribution information and the possibility to quantify metabolites. Accurate mass measurements proved to be critical in order to reliably combine data from different MS techniques. Initial results on the mycotoxin deoxynivalenol (DON) in wheat seed and phospholipids in mouse brain as a model for mammalian tissue indicate a broad applicability of the presented workflow. Copyright © 2014 John Wiley & Sons, Ltd.
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Mthiyane, Doctor Mziwenkosi Nhlanhla; Mhlanga, Bhekumusa Sabelo
2017-04-01
This study was aimed at investigating the nutritive value of marula seed cake (MSC) as an alternative protein source for broilers. In a completely randomised design involving six replicate pens of five chickens assigned to each of five treatments, body weight gain (BWG), feed intake (FI), feed conversion efficiency (FCE) and carcass characteristics were measured in an experiment in which 150 28-day-old broilers were fed maize-based diets containing, respectively, 0, 5, 10, 15 and 20% MSC at finisher phase. The results showed MSC to be remarkably high in CP (470.0 g/kg DM) and EE (343.5 g/kg DM), with moderate CF (58.2 g/kg DM), ash (54.3 g/kg DM), Ca (1.1 g/kg DM) and P (11.0 g/kg DM). Whilst very poor in lysine, MSC was found to be rich in methionine, cyst(e)ine, arginine and glutamic acid; it also contains good levels of valine, glycine, threonine, isoleucine, leucine, histidine, phenylalanine, serine, proline and alanine. Also, it contained 85.24% oleic (OA), 9.65% palmitic and 5.11% stearic acids, with a high peroxide value and low levels of mycotoxins deoxynivalenol (DON) and T-2 toxin. BWG, FI and FCE of broiler chickens significantly decreased (P < 0.001) as the dietary level of MSC increased. Further, dietary MSC significantly decreased bird live weight at slaughter (P < 0.001), plucked weight (P < 0.001), dressed weight (P < 0.001) and weights of the liver (P < 0.001) and neck (P < 0.05). The results therefore demonstrate MSC to be a good source of CP, fat, Ca, P, amino acids (except lysine) and OA that can replace soya bean meal (SBM) in broiler diets. However, its use is currently limited by lipid peroxidation and presence of mycotoxins.
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Yibadatihan, S; Jinap, S; Mahyudin, N A
2014-01-01
Palm kernel cake (PKC) is a useful source of protein and energy for livestock. Recently, it has been used as an ingredient in poultry feed. Mycotoxin contamination of PKC due to inappropriate handling during production and storage has increased public concern about economic losses and health risks for poultry and humans. This concern has accentuated the need for the evaluation of mycotoxins in PKC. Furthermore, a method for quantifying mycotoxins in PKC has so far not been established. The aims of this study were therefore (1) to develop a method for the simultaneous determination of mycotoxins in PKC and (2) to validate and verify the method. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using an electrospray ionisation interface (ESI) in both positive- and negative-ion modes was developed for the simultaneous determination of aflatoxins (AFB₁, AFB₂, AFG₁ and AFG₂), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisins (FB₁ and FB₂), T-2 and HT-2 toxin in PKC. An optimum method using a 0.2 ml min⁻¹ flow rate, 0.2% formic acid in aqueous phase, 10% organic phase at the beginning and 90% organic phase at the end of the gradient was achieved. The extraction of mycotoxins was performed using a solvent mixture of acetonitrile-water-formic acid (79:20:1, v/v) without further clean-up. The mean recoveries of mycotoxins in spiked PKC samples ranged from 81% to 112%. Limits of detection (LODs) and limits of quantification (LOQs) for mycotoxin standards and PKC samples ranged from 0.02 to 17.5 μg kg⁻¹ and from 0.06 to 58.0 μg kg⁻¹, respectively. Finally, the newly developed method was successfully applied to PKC samples. The results illustrated the fact that the method is efficient and accurate for the simultaneous multi-mycotoxin determination in PKC, which can be ideal for routine analysis.
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Mycotoxins in Crude Building Materials from Water-Damaged Buildings
Tuomi, Tapani; Reijula, Kari; Johnsson, Tom; Hemminki, Kaisa; Hintikka, Eeva-Liisa; Lindroos, Outi; Kalso, Seija; Koukila-Kähkölä, Pirkko; Mussalo-Rauhamaa, Helena; Haahtela, Tari
2000-01-01
We analyzed 79 bulk samples of moldy interior finishes from Finnish buildings with moisture problems for 17 mycotoxins, as well as for fungi that could be isolated using one medium and one set of growth conditions. We found the aflatoxin precursor, sterigmatocystin, in 24% of the samples and trichothecenes in 19% of the samples. Trichothecenes found included satratoxin G or H in five samples; diacetoxyscirpenol in five samples; and 3-acetyl-deoxynivalenol, deoxynivalenol, verrucarol, or T-2-tetraol in an additional five samples. Citrinine was found in three samples. Aspergillus versicolor was present in most sterigmatocystin-containing samples, and Stachybotrys spp. were present in the samples where satratoxins were found. In many cases, however, the presence of fungi thought to produce the mycotoxins was not correlated with the presence of the expected compounds. However, when mycotoxins were found, some toxigenic fungi usually were present, even if the species originally responsible for producing the mycotoxin was not isolated. We conclude that the identification and enumeration of fungal species present in bulk materials are important to verify the severity of mold damage but that chemical analyses are necessary if the goal is to establish the presence of mycotoxins in moldy materials. PMID:10788357
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Mycotoxins in crude building materials from water-damaged buildings.
Tuomi, T; Reijula, K; Johnsson, T; Hemminki, K; Hintikka, E L; Lindroos, O; Kalso, S; Koukila-Kähkölä, P; Mussalo-Rauhamaa, H; Haahtela, T
2000-05-01
We analyzed 79 bulk samples of moldy interior finishes from Finnish buildings with moisture problems for 17 mycotoxins, as well as for fungi that could be isolated using one medium and one set of growth conditions. We found the aflatoxin precursor, sterigmatocystin, in 24% of the samples and trichothecenes in 19% of the samples. Trichothecenes found included satratoxin G or H in five samples; diacetoxyscirpenol in five samples; and 3-acetyl-deoxynivalenol, deoxynivalenol, verrucarol, or T-2-tetraol in an additional five samples. Citrinine was found in three samples. Aspergillus versicolor was present in most sterigmatocystin-containing samples, and Stachybotrys spp. were present in the samples where satratoxins were found. In many cases, however, the presence of fungi thought to produce the mycotoxins was not correlated with the presence of the expected compounds. However, when mycotoxins were found, some toxigenic fungi usually were present, even if the species originally responsible for producing the mycotoxin was not isolated. We conclude that the identification and enumeration of fungal species present in bulk materials are important to verify the severity of mold damage but that chemical analyses are necessary if the goal is to establish the presence of mycotoxins in moldy materials.
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Desjardins, Anne E; Busman, Mark; Manandhar, Gyanu; Jarosz, Andrew M; Manandhar, Hira K; Proctor, Robert H
2008-07-09
The fungus Fusarium graminearum (sexual stage Gibberella zeae) causes ear rot of maize (Zea mays) and contamination with the 8-ketotrichothecenes nivalenol (1) or 4-deoxynivalenol (2), depending on diversity of the fungal population for the 4-oxygenase gene (TRI13). To determine the importance of 1 and 2 in maize ear rot, a survey of naturally contaminated maize in Nepal was combined with experiments in the field and in a plant growth room. In the survey, 1 contamination was 4-fold more frequent than 2 contamination and 1-producers (TRI13) were isolated more than twice as frequently as 2-producers (Psi TRI13). In maize ear rot experiments, genetically diverse 1-producers and 2-producers caused ear rot and trichothecene contamination. Among strains with the same genetic background, however, 1-producers caused less ear rot and trichothecene contamination than did 2-producers. The high frequency of 1 contamination and the high virulence of many 1-producers are of concern because maize is a staple food of rural populations in Nepal and because 1 has proven to be more toxic than 2 to animals.
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Mycotoxins transmitted into beer from contaminated grains during brewing.
Scott, P M
1996-01-01
Studies with aflatoxin B1, ochratoxin A, zearalenone, deoxynivalenol, and fumonisins B1 and B2 added at various stages of the brewing process show that these mycotoxins (or metabolites) may be transmitted from contaminated grains into beer. Citrinin does not survive the mashing step. Mycotoxins in beer could originate from the malted grain or from adjuncts. Although high incidences and concentrations of aflatoxins and zearalenone have been found in local beers brewed in Africa, aflatoxins have not been detected in European beers. Zearalenone and alpha- or beta-zearalenol (the metabolite likely to occur) have not been found in Canadian and European beers, except for one sample analyzed by thin-layer chromatography only. Ochratoxin A rarely has been detected at > 1 ng/mL in beer; liquid chromatographic methods with a 0.05-0.1 ng/mL detection limit, however, have shown moderately high incidences of trace levels. Deoxynivalenol, which survives the brewing process, has been found with high incidence in Canadian and European beers, with concentration of > 200 ng/mL reported in several German beers. Fumonisins B1 and B2 occur to a limited extent in beer.
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Gajęcka, Magdalena; Stopa, Ewa; Tarasiuk, Michał; Zielonka, Łukasz; Gajęcki, Maciej
2013-01-01
The aim of the study was to verify the hypothesis that intoxication with low doses of mycotoxins leads to changes in the mRNA expression levels of nitric oxide synthase-1 and nitric oxide synthase-2 genes in tissues of the gastrointestinal tract and the liver. The experiment involved four groups of immature gilts (with body weight of up to 25 kg) which were orally administered zearalenone in a daily dose of 40 μg/kg BW (group Z, n = 18), deoxynivalenol at 12 μg/kg BW (group D, n = 18), zearalenone and deoxynivalenol (group M, n = 18) or placebo (group C, n = 21) over a period of 42 days. The lowest mRNA expression levels of nitric oxide synthase-1 and nitric oxide synthase-2 genes were noted in the sixth week of the study, in particular in group M. Our results suggest that the presence of low mycotoxin doses in feed slows down the mRNA expression of both nitric oxide synthase isomers, which probably lowers the concentrations of nitric oxide, a common precursor of inflammation. PMID:24284830
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NASA Astrophysics Data System (ADS)
Wymore, A.; Rodriguez-Cardona, B.; Coble, A. A.; Potter, J.; Lopez Lloreda, C.; Perez Rivera, K.; De Jesus Roman, A.; Bernal, S.; Martí Roca, E.; Kram, P.; Hruska, J.; Prokishkin, A. S.; McDowell, W. H.
2016-12-01
Watershed nitrogen exports are often dominated by dissolved organic nitrogen (DON); yet, little is known about the role ambient DON plays in ecosystems. As an organic nutrient, DON may serve as either an energy source or as a nutrient source. One hypothesized control on DON is nitrate (NO3-) availability. Here we examine the interaction of NO3- and DON in streams across temperate forests, tropical rainforests, and Mediterranean and taiga biomes. Experimental streams also drain contrasting Critical Zones which provide gradients of vegetation, soil type and lithology (e.g. volcaniclastic, granitic, ultramafic, Siberian Traps Flood Basalt) in which to explore how the architecture of the Critical Zone affects microbial biogeochemical reactions. Streams ranged in background dissolved organic carbon (DOC) concentration (1-50 mg C/L) and DOC: NO3- ratios (10-2000). We performed a series of ecosystem-scale NO3- additions in multiple streams within each environment and measured the change in DON concentration. Results demonstrate that there is considerable temporal and spatial variation across systems with DON both increasing and decreasing in response to NO3- addition. Ecologically this suggests that DON can serve as both a nutrient source and an energy source to aquatic microbial communities. In contrast, DOC concentrations rarely changed in response to NO3- additions suggesting that the N-rich fraction of the ambient dissolved organic matter pool is more bioreactive than the C-rich fraction. Contrasting responses of the DON and DOC pools indicate different mechanisms controlling their respective cycling. It is likely that DON plays a larger role in ecosystems than previously recognized.
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Fate of dissolved organic nitrogen in two stage trickling filter process.
Simsek, Halis; Kasi, Murthy; Wadhawan, Tanush; Bye, Christopher; Blonigen, Mark; Khan, Eakalak
2012-10-15
Dissolved organic nitrogen (DON) represents a significant portion of nitrogen in the final effluent of wastewater treatment plants (WWTPs). Biodegradable portion of DON (BDON) can support algal growth and/or consume dissolved oxygen in the receiving waters. The fate of DON and BDON has not been studied for trickling filter WWTPs. DON and BDON data were collected along the treatment train of a WWTP with a two-stage trickling filter process. DON concentrations in the influent and effluent were 27% and 14% of total dissolved nitrogen (TDN). The plant removed about 62% and 72% of the influent DON and BDON mainly by the trickling filters. The final effluent BDON values averaged 1.8 mg/L. BDON was found to be between 51% and 69% of the DON in raw wastewater and after various treatment units. The fate of DON and BDON through the two-stage trickling filter treatment plant was modeled. The BioWin v3.1 model was successfully applied to simulate ammonia, nitrite, nitrate, TDN, DON and BDON concentrations along the treatment train. The maximum growth rates for ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria, and AOB half saturation constant influenced ammonia and nitrate output results. Hydrolysis and ammonification rates influenced all of the nitrogen species in the model output, including BDON. Copyright © 2012 Elsevier Ltd. All rights reserved.
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-12-16
... DEPARTMENT OF COMMERCE Bureau of Industry and Security Donald V. Bernardo, a/k/a Don Bernarndo... the U.S. District Court, Southern District of Florida, Donald V. Bernardo, a/k/a Don Bernardo.... Accordingly, it is hereby ordered I. Until November 16, 2016, Donald V. Bernardo, a/k/a Don Bernardo, with a...
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Sun, Jingyi; Simsek, Halis
2017-07-01
Effluent dissolved organic nitrogen (DON) is problematic in nutrient sensitive surface waters and needs to be reduced to meet demanding total dissolved nitrogen discharge limits. Bioavailable DON (ABDON) is a portion of DON utilized by algae or algae+bacteria, while biodegradable DON (BDON) is a portion of DON decomposable by bacteria. ABDON and BDON in a two-stage trickling filter (TF) wastewater treatment plant was evaluated using three different microalgal species, Selenastrum capricornutum, Chlamydomonas reinhardtii and Chlorella vulgaris and mixed cultured bacteria. Results showed that up to 80% of DON was bioavailable to algae or algae+bacteria inoculum while up to 60% of DON was biodegradable in all the samples. Results showed that C. reinhardtii and C. vulgaris can be used as a test species the same as S. capricornutum since there were no significant differences among these three algae species based on their ability to remove nitrogen species. Copyright © 2017. Published by Elsevier B.V.
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The Don'ts and Don'ts of Teaching
ERIC Educational Resources Information Center
Rubinstein, Gary
2012-01-01
Gary Rubinstein doesn't buy the adage that rules--for students or for teachers--must be stated positively, without don'ts or nots. So he lists 10 rules for things teachers must not do in their first years. Tops on his list is "Don't try to teach too much in one day," which Rubinstein says is a common mis-step because teacher preparation programs,…
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Carbon and nitrogen stoichiometry across stream ecosystems
NASA Astrophysics Data System (ADS)
Wymore, A.; Kaushal, S.; McDowell, W. H.; Kortelainen, P.; Bernhardt, E. S.; Johnes, P.; Dodds, W. K.; Johnson, S.; Brookshire, J.; Spencer, R.; Rodriguez-Cardona, B.; Helton, A. M.; Barnes, R.; Argerich, A.; Haq, S.; Sullivan, P. L.; López-Lloreda, C.; Coble, A. A.; Daley, M.
2017-12-01
Anthropogenic activities are altering carbon and nitrogen concentrations in surface waters globally. The stoichiometry of carbon and nitrogen regulates important watershed biogeochemical cycles; however, controls on carbon and nitrogen ratios in aquatic environments are poorly understood. Here we use a multi-biome and global dataset (tropics to Arctic) of stream water chemistry to assess relationships between dissolved organic carbon (DOC) and nitrate, ammonium and dissolved organic nitrogen (DON), providing a new conceptual framework to consider interactions between DOC and the multiple forms of dissolved nitrogen. We found that across streams the total dissolved nitrogen (TDN) pool is comprised of very little ammonium and as DOC concentrations increase the TDN pool shifts from nitrate to DON dominated. This suggests that in high DOC systems, DON serves as the primary source of nitrogen. At the global scale, DOC and DON are positively correlated (r2 = 0.67) and the average C: N ratio of dissolved organic matter (molar ratio of DOC: DON) across our data set is approximately 31. At the biome and smaller regional scale the relationship between DOC and DON is highly variable (r2 = 0.07 - 0.56) with the strongest relationships found in streams draining the mixed temperate forests of the northeastern United States. DOC: DON relationships also display spatial and temporal variability including latitudinal and seasonal trends, and interactions with land-use. DOC: DON ratios correlated positively with gradients of energy versus nutrient limitation pointing to the ecological role (energy source versus nutrient source) that DON plays with stream ecosystems. Contrary to previous findings we found consistently weak relationships between DON and nitrate which may reflect DON's duality as an energy or nutrient source. Collectively these analyses demonstrate how gradients of DOC drive compositional changes in the TDN pool and reveal a high degree of variability in the C: N ratio (3-100) of stream water dissolved organic matter.
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Oboh, Ganiyu; Ogunsuyi, Opeyemi Babatunde; Olonisola, Oluwaseyi Emmanuel
2017-04-01
Caffeine is adjudged world's most consumed pharmacologically active food component. With reports of the potential cognitive enhancing properties of caffeine, we sought to investigate if caffeine can influence the anticholinesterase and antioxidant properties of donepezil-a selective acetylcholinesterase (AChE) inhibitor used in the management of Alzheimer's disease (AD). In vitro, we investigated the effect of donepezil (DON), caffeine (CAF) and their various combinations on the activity of AChE in rat brain homogenate, as well as determined their antioxidant properties. In vivo, two rat groups were administered single oral dose of DON (5 mg/kg) and CAF (5 mg/kg) separately, while three groups, each received 5 mg/kg DON plus either 5, 50 or 100 mg/kg CAF for three hours, after which the rats were sacrificed and brain isolated. Results show that CAF concentration dependently and synergistically increased the anticholinesterase properties of DON in vitro. Also, CAF produced a significant influence on investigated in vitro antioxidant properties of DON. Furthermore, rats administered 5 mg/kg CAF and DON produced no significant difference in AChE activity compared to rats administered DON alone. However, co-administration of either 50 or 100 mg/kg CAF with DON lead to higher AChE activity compared to both control and DON groups. In addition, DON, CAF and their various combinations augmented brain antioxidant status in treated rats. We conclude that while low caffeine consumption may improve the antioxidant properties of donepezil without having a significant influence on its anticholinesterase effect, moderate-high caffeine consumption could also improve the antioxidant properties of donepezil but reduce its anticholinesterase effect; nevertheless, a comprehensive clinical trial is essential to fully explore these possibilities in human AD condition.
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NASA Astrophysics Data System (ADS)
Paerl, H. W.; Peierls, B. L.; Hounshell, A.; Osburn, C. L.
2015-12-01
Eutrophication is a widespread problem affecting the structure and function of estuaries and is often linked to anthropogenic nitrogen (N) enrichment, since N is the primary nutrient limiting algal production. Watershed management actions typically have ignored dissolved organic nitrogen (DON) loading because of its perceived refractory nature and instead focused on inorganic N as targets for loading reductions. A fluorescence-based model indicated that anthropogenic sources of DON near the head of the microtidal Neuse River Estuary (NRE), NC were dominated by septic systems and poultry waste. A series of bioassays were used to determine the bioavailability of river DON and DON-rich sources to primary producers and whether those additions promoted the growth of certain phytoplankton taxa, particularly harmful species. Overall, at time scales up to two to three weeks, estuarine phytoplankton and bacteria only showed limited responses to additions of high molecular weight (HMW, >1 kDa) river DON. When increases in productivity and biomass did occur, they were quite small compared with the response to inorganic N. Low molecular weight (LMW) river DON, waste water treatment plant effluent, and poultry litter extract did have a positive effect on phytoplankton and bacterial production, indicating a bioavailable fraction. High variability of bulk DON concentration suggested that bioavailable compounds added in the experimental treatments were low in concentration and turned over quite rapidly. Some phytoplankton taxa, as measured by diagnostic photopigments, appeared to be selectively enhanced by the HMW and specific source DON additions, although the taxa could not be positively identified as harmful species. Preliminary tests show that labile autochthonous organic matter may act as a primer for the mineralization of the HMW DON. These and other, longer-term bioavailability studies will be needed to adequately address the fate of watershed DON in estuarine ecosystems.
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Guidelines: the do's, don'ts and don't knows of feedback for clinical education.
Lefroy, Janet; Watling, Chris; Teunissen, Pim W; Brand, Paul
2015-12-01
The guidelines offered in this paper aim to amalgamate the literature on formative feedback into practical Do's, Don'ts and Don't Knows for individual clinical supervisors and for the institutions that support clinical learning. The authors built consensus by an iterative process. Do's and Don'ts were proposed based on authors' individual teaching experience and awareness of the literature, and the amalgamated set of guidelines were then refined by all authors and the evidence was summarized for each guideline. Don't Knows were identified as being important questions to this international group of educators which if answered would change practice. The criteria for inclusion of evidence for these guidelines were not those of a systematic review, so indicators of strength of these recommendations were developed which combine the evidence with the authors' consensus. A set of 32 Do and Don't guidelines with the important Don't Knows was compiled along with a summary of the evidence for each. These are divided into guidelines for the individual clinical supervisor giving feedback to their trainee (recommendations about both the process and the content of feedback) and guidelines for the learning culture (what elements of learning culture support the exchange of meaningful feedback, and what elements constrain it?) Feedback is not easy to get right, but it is essential to learning in medicine, and there is a wealth of evidence supporting the Do's and warning against the Don'ts. Further research into the critical Don't Knows of feedback is required. A new definition is offered: Helpful feedback is a supportive conversation that clarifies the trainee's awareness of their developing competencies, enhances their self-efficacy for making progress, challenges them to set objectives for improvement, and facilitates their development of strategies to enable that improvement to occur.