Trichothecene Mycotoxins Research Articles

Impact of nitrogen metabolism-associated culture pH changes on regulation of Fusarium trichothecene biosynthesis: revision of roles of polyamine agmatine and transcription factor AreA.

Fusarium graminearum produces trichothecene mycotoxins in infected grains and axenic liquid culture. A proposed regulatory model of trichothecene biosynthesis was examined in relation to nitrogen utilization. First, we showed that an important factor for the stimulation of trichothecene biosynthesis was not the occurrence of agmatine as a specific inducer molecule, but rather continuous acidification of the liquid culture medium arising from agmatine catabolism. When the pH of the L-Gln synthetic medium was frequently adjusted to the pH of the agmatine culture, trichothecene productivity of the L-Gln culture was equal to that of the agmatine culture. For efficient trichothecene biosynthesis, the culture pH should be lowered at an appropriate time point during the early growth stage. Second, we re-evaluated the role of the nitrogen regulatory GATA transcription factor AreA in trichothecene biosynthesis. Since Tri6 encodes a transcription factor indispensable for trichothecene biosynthesis, all fifteen AreA-binding consensus sequences in the Tri6 promoter were mutated. The mutant could catabolize L-Phe as the sole nitrogen source; furthermore, the pH profile of the synthetic L-Phe medium (initial pH 4.2) was the same as that of the wild-type (WT) strain. Under such conditions, the promoter mutant exhibited approximately 72% of the trichothecene productivity compared to the WT strain. Thus, F. graminearum AreA (FgAreAp) is dispensable for the functioning of the Tri6 promoter, but it contributes to the increased production of mycotoxin under mildly acidic conditions to some extent. Further investigations on the culture pH revealed that extremely low pH bypasses the function of FgAreAp.

Suppression of Trichothecene-Mediated Immune Response by the Fusarium Secondary Metabolite Butenolide in Human Colon Epithelial Cells.

Butenolide (BUT, 4-acetamido-4-hydroxy-2-butenoic acid gamma-lactone) is a secondary metabolite produced by several Fusarium species and is co-produced with the major trichothecene mycotoxin deoxynivalenol (DON) on cereal grains throughout the world. BUT has low acute toxicity and only very limited occurrence and exposure data are available. The intestinal epithelium represents the first physiological barrier against food contaminants. We aimed to elucidate the intestinal inflammatory response of the human, non-cancer epithelial HCEC-1CT cells to BUT and to characterize potential combinatory interactions with co-occurring trichothecenes, such as DON and NX-3. Using a reporter gene approach, BUT (≥5 μM, 20 h) was found to decrease lipopolysaccharide (LPS; 10 ng/mL) induced nuclear factor kappa B (NF-κB) activation in a dose-dependent manner, and in combinatory treatments BUT represses trichothecene-induced enhancement of this important inflammatory pathway. Analysis of transcription and secretion levels of NF-κB-dependent, pro-inflammatory cytokines, revealed a significant down-regulation of IL-1β, IL-6, and TNF-α in IL-1β-stimulated (25 ng/mL) HCEC-1CT cells after BUT exposure (10 μM). Trichothecene-induced expression of pro-inflammatory cytokines by the presence of 1 μM DON or NX-3 was substantially suppressed in the presence of 10 μM BUT. The emerging mycotoxin BUT has the ability to suppress NF-κB-induced intestinal inflammatory response mechanisms and to modulate substantially the immune responsiveness of HCEC-1CT cells after trichothecene treatment. Our results suggest that BUT, present in naturally occurring mixtures of Fusarium fungal metabolites, should be increasingly monitored, and the mechanism of inhibition of NF-κB that might affect the pathogenesis or progression of intestinal inflammatory disorders, should be further investigated.

Low-level contamination of deoxynivalenol: A threat from environmental toxins to porcine epidemic diarrhea virus infection

Mycotoxins are toxic metabolites produced by fungal species that commonly present in the global environment, especially in cereals and animal forages. The changing global environment may further increase the exposure to these toxins, posing a serious threat to humans and animals. Recently, coronavirus has become one of the most important pathogens threatening human and animal health. It is not clear whether environmental toxins, such as mycotoxins, will affect coronavirus infection. Given that pigs are among the animals most affected by coronavirus and highly homologous to humans, weaned piglets and IPEC-J2 cells were respectively chosen as in vivo and in vitro model to explore the impacts of deoxynivalenol (DON), the most abundant trichothecene mycotoxin in feed, on porcine epidemic diarrhea virus (PEDV) infection and the mechanisms involved. In vivo, twenty-seven piglets infected naturally with PEDV were randomly divided into three groups, receiving the basal diet containing 0, 750 and 1500 μg/kg DON, respectively. Significant increases in the diarrhea rates, gut barrier injury and PEDV proliferation of piglets’ small intestine were observed in experimental groups compared with the control. Additionally, the autophagosome-like vesicles and the autophagy-related proteins expression were also increased in experimental groups. In vitro, we observed that 0.1, 0.5 and 1.0 μM DON significantly promoted the entry and replication of PEDV in IPEC-J2 cells, along with the induction of a complete autophagy. CRISPR‐Cas9‐mediated knockout of LC3B indicated a vital role of autophagy in the promotion. Pretreatment with p38 signaling inhibitor could significantly block the induction of autophagy, indicating that DON could promote the PEDV infection by triggering p38-mediated autophagy. Our findings suggest that mycotoxin could influence the prevalence of coronavirus and provide new ideas for the prevention and control of coronavirus.

Type A Trichothecene Diacetoxyscirpenol-Induced Emesis Corresponds to Secretion of Peptide YY and Serotonin in Mink

The trichothecene mycotoxins contaminate cereal grains and have been related to alimentary toxicosis resulted in emetic response. This family of mycotoxins comprises type A to D groups of toxic sesquiterpene chemicals. Diacetoxyscirpenol (DAS), one of the most toxic type A trichothecenes, is considered to be a potential risk for human and animal health by the European Food Safety Authority. Other type A trichothecenes, T-2 toxin and HT-2 toxin, as well as type B trichothecene deoxynivalenol (DON), have been previously demonstrated to induce emetic response in the mink, and this response has been associated with the plasma elevation of neurotransmitters peptide YY (PYY) and serotonin (5-hydroxytryptamine, 5-HT). However, it is found that not all the type A and type B trichothecenes have the capacity to induce PYY and 5-HT. It is necessary to identify the roles of these two emetogenic mediators on DAS-induced emesis. The goal of this study was to determine the emetic effect of DAS and relate this effect to PYY and 5-HT, using a mink bioassay. Briefly, minks were fasted one day before experiment and given DAS by intraperitoneally and orally dosing on the experiment day. Then, emetic episodes were calculated and blood collection was employed for PYY and 5-HT test. DAS elicited robust emetic responses that corresponded to upraised PYY and 5-HT. Blocking the neuropeptide Y2 receptor (NPY2R) diminished emesis induction by PYY and DAS. The serotonin 3 receptor (5-HT3R) inhibitor granisetron totally restrained the induction of emesis by serotonin and DAS. In conclusion, our findings demonstrate that PYY and 5-HT have critical roles in DAS-induced emetic response.

N-acetyl-cysteine in combination with celecoxib inhibits Deoxynivalenol induced skin tumor initiation via induction of autophagic pathways in swiss mice

Deoxynivalenol is a trichothecene mycotoxin which naturally contaminates small grain, cereals intended for human and animal consumption. Investigations for dermal toxicity of DON has been needed and highlighted by WHO. Previous studies on dermal toxicity suggest that DON has DNA damaging potential leading to skin tumor initiation in mice skin. However, considering its toxicological manifestations arising after dermal exposure, strategies for its prevention/protection are barely available in literatute. Collectively, our study demonstrated that N-acetylcysteine (NAC), precursor of glutathione, significantly alters the genotoxic potential of DON. Further NAC in combination with Celecoxib (CXB) inhibits tumor growth by altering antioxidant status and increasing autophagy in DON initiated Swiss mice. Despite the broad spectrum use of CXB, its use is limited by the concerns about its adverse effects on the cardiovascular system. Serum parameters and histology analysis revealed that CXB (2 mg) when applied topically for 24 weeks did not impart any cardiovascular toxicity which could be because skin permeation potential of CXB was quite low when analyzed through HPLC analysis.Although the anticancer effects of CXB and NAC have been studied, however, the combination of NAC and CXB has yet not been explored for any cancer treatment. Therefore our observations provide additional insights into the therapeutic effects of combinatorial treatment of CXB and NAC against skin tumor prevention. This approach might form a novel alternative strategy for skin cancer treatment as well as skin associated toxicities caused by mycotoxins such as DON. This combinatorial approach can overcome the limitations associated with the use of CXB for long term as topical application of the same seems to be safe in comparison to the oral mode of administration.

Synbiotic (PoultryStar® Sol) Protects Rat Liver from Oxidoreductive Stress Induced by T2-Mycotoxicity

Mycotoxins are secondary metabolic byproducts of molds that posses harmful and detrimental impact on animal and human health. Fungal T2 mycotoxin is the most efficacious trichothecene mycotoxin. It can induce hepatic and intestinal damages through eliciting severe oxidoreductive stress. Consequently, dysbiosis (alteration in the compositional balance of intestinal microbiota) occurred that can play an important role in the pathogenesis of several liver diseases. Recently, the using of pro and prebiotics was proven to exert antioxidative stress potentials in liver and gut ecosystem. To explore, the efficacy of using synbiotic (PoultryStar® Sol) during chronic T-2 mycotoxicosis course in rat, this study was designed. Rats were allocated into 4 main groups, (CN-Gr), which was treated as the negative control and was allowed for the free access to the normal rats chow and the tap water for 90 days, (PSS-Gr), which was assigned as the synbiotic positive control group and was allowed for the free access to normal rat chow and (PoultryStar® Sol) in the tap water (20mg/rat every 5 days) for 90 days. Group 3 or (T2-Gr) and was given the T2 contaminated chow ad libitumat a level of (470 ppb) along with normal drinking water for 90 days, and Group 4 (T2+PSS-Gr) which was given the T-2 contaminated chow ad libitumat the same level of Group 3 along with synbiotic (PoultryStar® Sol) in the tap water the same dose of (PSS-Gr) for 90 days. Each one of the four groups was subdivided into two subgroups (n=7) that one was exsanguinated on day 45 (Subgroup A) meantime the remaining rats (Subgroup B) were kept until the end of the entire study period (90 days). In T2 intoxicated group, liver histopathological findings on day 45 revealed lesions of reversible types (hydropic and fatty degenerations) only. Concurrently, on day 90 the changes comprised apoptosis and some necroinflammatory lesions. In T2+PSS-Gr, the ultimate efficacy of synbiotic success in preventing hepatic necroinflammatory changes and limiting the apoptotic activity which were in accordance with the amelioration of antioxidative status, liver function enzymes and other serum biochemical estimated tests. Summing up, the total results of this study has been clarify the regenerative and antioxidant potentials of (PoultryStar® Sol) in coping with T2 toxin mediated hepatotoxicity in rats.

The mycotoxin deoxynivalenol activates GABAergic neurons in the reward system and inhibits feeding and maternal behaviours

Deoxynivalenol (DON) or vomitoxin, is a trichothecene mycotoxin produced mainly by Fusarium graminearum and culmorum. Mycotoxins or secondary metabolic products of mold fungi are micro-pollutants, which may affect human and animal health. The neuronal and behavioural actions of DON were analysed in the present study. To address, which neurons can be affected by DON, the neuronal activation pattern following intraperitoneal injection of DON (1 mg/kg) was investigated in adult male rats and the results were confirmed in mice, too. DON-induced neuronal activation was assessed by c-Fos immunohistochemistry. DON injection resulted in profound c-Fos activation in only the elements of the reward system, such as the accumbens nucleus, the medial prefrontal cortex, and the ventral tegmental area. Further double labelling studies suggested that GABAergic neurons were activated by DON treatment. To study the behavioural relevance of this activation, we examined the effect of DON on feed intake as an example of reward-driven behaviours. Following DON injection, feed consumption was markedly reduced but returned to normal the following day suggesting an inhibitory action of DON on feed intake without forming taste-aversion. To further test how general the effect of DON on goal-directed behaviours is, its actions on maternal behaviour was also examined. Pup retrieval latencies were markedly increased by DON administration, and DON-treated mother rats spent less time with nursing suggesting reduced maternal motivation. In a supplementary control experiment, DON did not induce conditioned place preference arguing against its addictive or aversive actions. The results imply that acute uptake of the mycotoxin DON can influence the reward circuit of the brain and exert inhibitory actions on goal-directed, reward-driven behaviours. In addition, the results also suggest that DON exposure of mothers may have specific implications.

Effects of Atmospheric-Pressure Cold Plasma Treatment on Deoxynivalenol Degradation, Quality Parameters, and Germination of Barley Grains

Deoxynivalenol (DON) is one of the major trichothecene mycotoxins commonly found in grains, in particular barley. This study focused on the reduction of DON concentration on barley samples using atmospheric cold plasma (ACP) treatment. The effects of moisture content, post-treatment storage, and relative humidity of air on DON degradation on barley were evaluated. Additionally, the germination and the quality parameters of barley, including protein content, β-glucan, and moisture content, were evaluated. The results showed that ACP treatment for 6 and 10 min reduced DON concentration by 48.9% and 54.4%, respectively. No significant differences were observed in the DON degradation levels by increasing the moisture content of barley from 9.5 to 15.7 g water/100 g sample and relative humidity of air from 12 to 60%. Steeping of barley grains without subsequent drying prior to ACP treatment significantly increased the degradation rate of DON by ACP due to the presence of water on the grain surface. No significant differences were observed for the tested quality parameters of barley in comparison with control samples. This study shows that ACP may offer an effective DON reduction in barley without affecting the quality attributes. However, ACP treatment parameters should be optimized to achieve a better DON reduction efficacy.

Changes in Wheat Nutritional Content at Elevated [CO2] Alter Fusarium graminearum Growth and Mycotoxin Production on Grain.

Rising atmospheric [CO2] has been shown to impact plant primary metabolism and the severity of Fusarium head blight (FHB) in wheat. In this study, we evaluated how changes in grain nutritional content due to growth at elevated [CO2] affected Fusarium graminearum growth and mycotoxin production. Susceptible (Norm) and moderately resistant (Alsen) hard spring wheat grains that had been grown at ambient (400 ppm) or elevated [CO2] (800 ppm) were independently inoculated with two F. graminearum fungal strains, which produce the trichothecene mycotoxin, deoxynivalenol. Under higher [CO2], FHB-susceptible and moderately resistant wheat had disproportionate losses in protein and mineral contents, with Alsen being more severely impacted. Furthermore, the F. graminearum strain 9F1 had increased mycotoxin biosynthesis in response to the loss of wheat nutritional content in Alsen. Our results demonstrate that future [CO2] conditions may provide a strain-specific pathogenic advantage on hosts, with greater losses in nutritional content.

ОПТИМИЗАЦИЯ ПРОТОКОЛА ИММУНОФЕРМЕНТНОГО АНАЛИЗА ДЛЯ ИНДИКАЦИИ Т-2 ТОКСИНА

T-2 toxin is one of the most common and toxic trichothecene mycotoxins – secondary metabolites of molds that develop on cereals and some other crops. This article discusses the development stage of a test system for the determination of T-2 toxin based on enzyme-linked immunosorbent assay (ELISA), which uses an enzyme label – a conjugate of anti-rabbit goat antibodies with peroxidase. An important step in the creation of any ELISA-based test system is the preliminary titration of reaction components. The aim of the work was to determine the optimal concentration of the conjugate of antispecies antibodies in an indirectly competitive enzyme-linked immunosorbent enzyme immunoassay with the indication of T-2 toxin. A number of dilutions of the antivirus conjugate were examined: 1 : 1000; 1 : 2000; 1 : 3000; 1 : 4000; 1 : 5000; 1 : 7500; 1 : 10000; 1 : 12 500; 1 : 15,000; 1 : 17,500; 1 : 20,000; 1 : 30 000. In the ELISA staging protocol, calibration solutions of T-2 toxin at concentrations of 0.0 were used; 2.5; 5, 10, 20, 40, 80, 160 ng/ml and specific polyclonal rabbit antibodies to the T-2-BSA conjugate. Based on the average optical density values, calibration plots were constructed using the percentage of signal absorption from the zero standard. When evaluating the results of the study, the criterion for choosing the dilution of the conjugate of anti-species antibodies was considered the greatest, at which the best linearity of the grading plot is achieved and the level of its non-specific reaction with the zero standard would be the lowest. It was established that the optimal variants of dilutions of the conjugate of anti-species antibodies under the same experimental conditions tested were 1: 4000, 1 : 5000 and 1 : 12 500. Dose-dependent signal absorption was observed in all concentrations of anti-species antibodies. Dilutions of the conjugate of anti-species antibodies 1 : 1000–1 : 3000 and 1 : 17 500–1 : 30 000 were not taken into account, since the optical density of most wells was higher than the optimal boundaries in the first case (> 3.9) and lower in the second (< 0.4). Based on the foregoing, optimal dilution of the conjugate of anti-species antibodies was selected 1 : 12 500.

Intestinal Metabolism of α- and β-Glucosylated Modified Mycotoxins T-2 and HT-2 Toxin in the Pig Cecum Model.

The type A trichothecene mycotoxins T-2 and HT-2 toxin are fungal secondary metabolites produced by Fusarium fungi, which contaminate food and feed worldwide. Especially as a result of the high toxicity of T-2 toxin and their occurrence together with glucosylated forms in cereal crops, these mycotoxins are of human health concern. Particularly, it is unknown whether and how these modified mycotoxins are metabolized in the gastrointestinal tract and, thus, contribute to the overall toxicity. Therefore, the comparative intestinal metabolism of T-2 and HT-2 toxin glucosides in α and β configuration was investigated using the ex vivo pig cecum model, which mimics the human intestinal metabolism. Regardless of its configuration, the C-3 glycosidic bond was hydrolyzed within 10-20 min, releasing T-2 and HT-2 toxin, which were further metabolized to HT-2 toxin and T-2 triol, respectively. We conclude that T-2 and HT-2 toxin should be evaluated together with their modified forms for risk assessment.

Effects of maternal T-2 toxin exposure on the hepatic glycolipid metabolism in young mice.

T-2 toxin is a kind of group A trichothecenes mycotoxins, frequently detected in various foods and feeds, having high toxic effects on both humans and animals. The present study aims to investigate the toxic effects of T-2 toxin exposure to ICR mice during pregnancy and lactation on liver glycolipid metabolism of young mice. The pregnant mice were given 0, 0.005 and 0.05mg of T-2 toxin/kg bw daily through oral gavage from late gestation (GD 14) to the lactation (LD 21). Liver and serum samples of the young mice were collected on postnatal day 21 (PND 21), PND 28 and PND 56. The results showed that T-2 toxin increased the contents of triglycerides (TG), total cholesterol (T-CHO) and glucose in serum of young mice on PND 21 and PND 28. In addition, obvious lipid droplets of liver in T-2 toxin treatment groups were observed, especially in 0.05mg group of PND 21and PND 28. Compared with the control group, T-2 treatment also increased the expressions of genes associated with liver glycolipid metabolism, such as PEPCK, Glut2, Fas, Acox1, Hmgcr, PPARα, Srebp1 and CD36. These results demonstrated T-2 toxin exposure to pregnant mice could cause liver glycolipid metabolism disruption in the young mice and the toxic effects weakened on PND 56.

The role and regulatory mechanism of autophagy in hippocampal nerve cells of piglet damaged by deoxynivalenol

Deoxynivalenol (DON), a type B trichothecene mycotoxin mainly affects the health status of pigs and reduced their growth. This study aimed to determine the effects of PI3K/Akt/mTOR pathway on DON-induced autophagy of piglet hippocampal nerve cells (PHNCs), and the relationship between autophagy and apoptosis. The effects of DON on autophagy of PHNCs were examined by cell morphology, cell viability, apoptosis rate, electron microscopy, transient transfection of GFP-LC3 plasmid, immunofluorescence and expression of autophagy-related genes and proteins. The relationship between autophagy and cell apoptosis was analyzed by western blotting, CCK-8 and flow cytometry. The results indicated that, DON inhibited the proliferation of PHNCs and significantly changed cell morphology, and induced apoptosis and autophagy. The expression levels of LC3 protein and gene increased, while the expression levels of PI3K/Akt/mTOR pathway-related genes and proteins decreased, when the concentration of DON increased. Activation of autophagy significantly increased cell viability, reduced apoptosis rate, inhibits autophagy significantly, reduced cell activity and increased apoptosis rate. This data demonstrated that DON exerts certain toxic effect on PHNCs, induced apoptosis and autophagy. PI3K/Akt/mTOR signaling pathway plays a negative regulatory role in DON-induced autophagy of PHNCs. At the same time, autophagy plays a protective role in DON-induced PHNCs injury.

Enhanced Resistance toFusarium graminearumin Transgenic Arabidopsis Plants Expressing a Modified Plant Thionin

The fungal pathogen Fusarium graminearum causes Fusarium head blight (FHB) on wheat, barley, and other grains. FHB results in yield reductions and contaminates grain with trichothecene mycotoxins, which threaten food safety and food security. Innovative mechanisms for controlling FHB are urgently needed. We have previously shown that transgenic tobacco and citrus plants expressing a modified thionin (Mthionin) exhibited enhanced resistance toward several bacterial pathogens. The aim of this study was to investigate whether overexpression of Mthionin could be similarly efficacious against F. graminearum, and whether transgenic expression of Mthionin impacts the plant microbiome. Transgenic Arabidopsis plants expressing Mthionin were generated and confirmed. When challenged with F. graminearum, Mthionin-expressing plants showed less disease and fungal biomass in both leaves and inflorescences compared with control plants. When infiltrated into leaves, macroconidia of F. graminearum germinated at lower rates and produced less hyphal growth in Arabidopsis leaves expressing Mthionin. Moreover, marker genes related to defense signaling pathways were expressed at significantly higher levels after F. graminearum infection in Mthionin transgenic Arabidopsis plants. However, Mthionin expression did not appreciably alter the overall microbiome associated with transgenic plants grown under controlled conditions; across leaves and roots of Mthionin-expressing and control transgenic plants, only a few bacterial and fungal taxa differed, and differences between Mthionin transformants were of similar magnitude compared with control plants. In sum, our data indicate that Mthionin is a promising candidate to produce transgenic crops for reducing FHB severity and ultimately mycotoxin contamination.

Maternal Exposure to T-2 Toxin Affects Puberty Genes and Delays Estrus Cycle in Mice Offspring.

Simple SummaryThe experiment was designed to investigate the effect of gestational and lactational exposure to the T-2 toxin and its effects on the puberty of female mice offspring. T-2 toxin contaminates feed participates in the maternal diet. It directly interferes with the normal physiological function of offspring because the maternal diet is the most important during pregnancy and lactation. Female mice are more sensitive to the maternal diet during the lactation stage. Thus, their maturity stage is very complex and regulated by the hypothalamus, which controls Gonadotropin-releasing hormone (GnRH), and it also secretes reproductive hormones. Current findings report that T-2 toxins during late gestation and lactation period directly affect offspring, disturb the vaginal environment, delayed normal estrus cycle, and puberty age induces oxidative stress-caused ovarian damage. Based on findings, we think it is necessary to raise people’s attention to T-2 toxin-contaminated feed offered to animals. Current results reveal information about the health risks associated with a maternal diet, which affects the next generation.Among foodborne toxicities, the T-2 toxin is the most toxic member of trichothecenes mycotoxins, which has been shown to impair the development and reproductive efficiency of animals. Pups are particularly more quickly prone to programming the effects of the maternal diet during the gestational and lactation periods. Few studies have reported the maternal toxic effect on the next generation. Dams were served the T-2 toxin at a dose of 0.005 and 0.05 mg/kg body weight/day and control group 0 mg/kg from gestation day 14 to lactation day 21. Female mice offspring were selected at the weaning age. Our observations indicate that age during the vaginal opening and di-estrus stage increased and the length of the estrus cycle, first di-estrus, and regular estrus cycling were delayed with prolonged di-estrus in the 0.05 mg/kg group compared to the 0.005 mg/kg and control group. Transcription level analysis showed that mice at a dose of 0.05 mg/kg exhibited a decrease in hypothalamic mRNA expression of Gnrh and Gnrhr, Lhb, and Fshb in the pituitary gland, with a significant decrease of Fshr and Lhr in the ovaries. Present findings report that postnatal exposure to the T-2 toxin delayed puberty age in female mice and induced oxidative stress, ovarian damage, and reduced vaginal epithelium wall majorly in the 0.05 mg/kg group, and showed fewer effects in the 0.005 mg/kg group.

Anorexic responses to trichothecene deoxynivalenol and its congeners correspond to secretion of tumor necrosis factor-α and interleukin-1β

Type B trichothecene mycotoxins comprise deoxynivalenol (“Vomitoxin”, DON) and four structually related congeners: 15-acetyl- and 3-acetyl-deoxynivalenol (15-ADON and 3-ADON), nivalenol (NIV), 4-acetyl-nivalenol (fusarenon X, FX). These foodborne mycotoxins has been linked to food poisoning leading to anorexic response in human and several animal species. However, the pathophysiological basis for anorexic effect is relatively unclear. The goal of this research was to compare anorexic effect to type B trichothecenes and relate these effects to two common cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) following oral and IP exposure. Both cytokines were increased within 1−2 h in plasma and returned to basal concentrations at 6 h following exposure to DON and ADONs. FX evoked both cytokines with initial time and duration at 1−2 h and > 6 h, respectively. Elevation of TNF-α and IL-1β induced by orally exposure to NIV did not occur until 2 h and recovered to basal concentrations at 6 h. Both cytokines were elevated at 1 h and lasted more than 6 h following IP exposure to NIV. Type B trichothecenes stimulated plasma secretion of both cytokines that were consistent with reduction of food intake. In conclusion, our findings demonstrate that TNF-α and IL-1β act critical roles in type B trichothecenes-induced anorexic response.

Intrapopulation Antagonism Can Reduce the Growth and Aggressiveness of the Wheat Head Blight Pathogen Fusarium graminearum.

Fusarium graminearum is a causal agent of Fusarium head blight (FHB), a disease that reduces yield and quality of cereal crops and contaminates grain with mycotoxins that pose health risks to humans and livestock. Interpopulation antagonistic interactions between isolates that produce different trichothecene mycotoxins can reduce FHB in wheat, but it is not known if interactions between isolates with a shared population identity that produce the same trichothecenes have a similar effect. Using isolates from the predominant F. graminearum populations in North America (NA1 and NA2), we examined intrapopulation interactions by comparing growth, disease progression, and toxin production of individual isolates with multi-isolate mixes. In vitro, mycelial growth was significantly greater when most NA1 and NA2 isolates were cultured individually versus when cultured as a mixture of isolates from the same population. In susceptible wheat Norm, FHB generally progressed faster in heads inoculated with an individual isolate versus a multi-isolate mixture, but the antagonistic effect of intrapopulation interactions was more pronounced for NA1 than NA2 isolates. By contrast, in moderately resistant wheat Alsen, mixtures of isolates from either population caused obvious reductions in FHB development. Mycotoxin contamination was not consistently affected by intrapopulation interactions and varied depending on the interacting isolates from either population. Our results indicate that antagonistic intrapopulation interactions can influence FHB in controlled environmental conditions. Understanding if the regional composition of pathogen populations similarly influences FHB in the field could improve disease forecasting and management practices.

Occurrence of Deoxynivalenol, Nivalenol, and Their Glucosides in Korean Market Foods and Estimation of Their Population Exposure through Food Consumption.

Major type B trichothecene mycotoxins, including deoxynivalenol (DON), nivalenol (NIV), and their respective glucoside conjugates, deoxynivalenol-3-β-D-glucose (DON3G) and nivalenol-3-β-D-glucose (NIV3G), are present in food products, such as cereals, legumes, and their processed products. Thus, here, DON, NIV, and their 3-β-D-glucosides were monitored in 506 Korean market foods, and exposure to these mycotoxins was estimated in the population consuming these foods. The accuracy and precision of our method, which simultaneously determined four toxins, were 80.1–106.5% and 0.3–12.4%, in four representative food matrices assessed. The incidences of DON, DON3G, NIV, and NIV3G among all food samples tested were 13%, 8%, 12%, and 5%, respectively. The glucoside conjugate with free toxin was found to have the maximum co-occurrence of 49%. The estimated daily intakes of DON, DON3G, NIV, and NIV3G through food intake under four different scenarios were 0.019–0.102, 0.004–0.089, 0.007–0.094, and 0.002–0.095 μg kg−1 body weight (b.w.) day−1, respectively, which are lower than the established health-based guidance values. Overall, our results suggest that the estimated exposure of the Korean population to type B trichothecenes, namely, DON, NIV, and their 3-β-D-glucoside conjugates, may not pose a potential health risk.

T-2 Toxin Induces Oxidative Stress, Apoptosis and Cytoprotective Autophagy in Chicken Hepatocytes.

T-2 toxin is type A trichothecenes mycotoxin, which produced by fusarium species in cereal grains. T-2 toxin has been shown to induce a series of toxic effects on the health of human and animal, such as immunosuppression and carcinogenesis. Previous study has proven that T-2 toxin caused hepatotoxicity in chicken, but the regulatory mechanism is unclear. In the present study, we assessed the toxicological effect of T-2 toxin on apoptosis and autophagy in hepatocytes. The total of 120 1-day-old healthy broilers were allocated randomly into four groups and reared for 21 day with complete feed containing 0 mg/kg, 0.5 mg/kg, 1 mg/kg or 2 mg/kg T-2 toxin, respectively. The results showed that the apoptosis rate and pathological changes degree hepatocytes were aggravated with the increase of T-2 toxin. At the molecular mechanism level, T-2 toxin induced mitochondria-mediated apoptosis by producing reactive oxygen species, promoting cytochrome c translocation between the mitochondria and cytoplasm, and thus promoting apoptosomes formation. Meanwhile, the expression of the autophagy-related protein, ATG5, ATG7 and Beclin-1, and the LC3-II/LC3-I ratio were increased, while p62 was downregulated, suggesting T-2 toxin caused autophagy in hepatocytes. Further experiments demonstrated that the PI3K/AKT/mTOR signal may be participated in autophagy induced by T-2 toxin in chicken hepatocytes. These data suggest a possible underlying molecular mechanism for T-2 toxin that induces apoptosis and autophagy in chicken hepatocytes

Cardiomyopathy induced by T-2 toxin in rats

T-2 toxin, A trichothecenes mycotoxin, is immunotoxic to animals and humans. Although it is highly cardiotoxic, the pathogenesis of cardiomyopathy caused by T-2 toxin is not entirely clear. Hence, in our research, cardiomyopathy was induced by a single injection of T-2 mycotoxin (0.23 mg/kg s.c., 1 LD50) to Wistar rats. The cardiac tissue was carefully examinated by using basic histopathology, semiquantitative (tissue grading score scales) and imaging (a total number of mast cells – MCs) analyses on days 1, 7, 14, 21, 28 and 60 of the study. The most intensive myocardial alterations (cardiac damage score, CDS = 4.20–4.40), irregular glycogen distribution (glycogen distribution score, GDS = 4.07–4.17), haemorrhagic foci (vascular damage score, VDS = 4.57–4.90), diffuse accumulation and degranulation of MCs were observed on day 28 and 60 after treatment (p < 0.001 vs. control and 1st T-2-toxin-treated group, respectively). Besides, statistically significant positive correlations were obtained regarding myocardial injury, glycogen distribution and intensity of haemorrhage, and a negative correlation was found in the case of MCs. Obtained results are essential and crucial for further in vivo experimental studies, including the development of medications able to reduce T-2 toxin-induced cardiotoxicity.