Oral exposure to Nivalenol induced apoptosis in lymphoid tissues (thymus and spleen) of mice. The apoptotic rate in thymocytes and splenocytes was significantly increased compared to the control group. Histopathological examination showed structural damage in lymphoid organs, including thymic cortical atrophy and splenic white pulp depletion [2]

Male ICR mice (4 weeks old) were used in the experiment. Nivalenol was dissolved in distilled water. Mice were orally administered with Nivalenol at doses of 0.25, 0.5, and 1 mg/kg body weight once daily for 7 consecutive days. Control mice received an equal volume of distilled water. At the end of the exposure period, mice were sacrificed by cervical dislocation, and thymus and spleen were collected for further analysis [2]

Absorption, Distribution and Excretion
In mice, novofusarium oxychloride rapidly distributes to and is cleared from all tested tissues without significant accumulation in any organ. Following long-term oral administration of novofusarium oxychloride, the recoveries in the feces of male rats were: novofusarium oxychloride (7%), de-epoxy novofusarium oxychloride (80%), novofusarium oxychloride (1%), and de-epoxy novofusarium oxychloride (1%) in urine. To investigate the metabolic pathways of novofusarium oxychloride (NIV) and its 4-acetyl derivative (fusarium oxychloride-X, FX) in mice, mice were orally administered either 3H-FX or 3H-NIV. Mice given 3H-FX primarily excreted the radioactive material in their urine, while mice given 3H-NIV primarily excreted it in their feces. Plasma radioactivity peaked 30 or 60 minutes after administration of either (3)H-FX or (3)H-NIV. Compared to mice given (3)H-NIV, mice given (3)H-FX showed a 5-fold higher peak plasma concentration and a 10-fold higher area under the curve (AUC). These results clearly demonstrate that FX is absorbed more readily and efficiently from the gastrointestinal tract than NIV. High-performance liquid chromatography analysis of acetonitrile extracts from urine and feces showed that FX is rapidly metabolized to NIV after absorption from the gastrointestinal tract. In vitro incubation experiments with tissue homogenates and (3)H-FX indicated that the liver and kidneys are the main organs for the conversion of FX to NIV. Therefore, this study indicates that the higher oral toxicity of FX compared to NIV observed in mice and rats is due to the fact that FX is more readily absorbed from the gastrointestinal tract than NIV, and subsequently rapidly converted to NIV in the liver and kidneys. Metabolites/Metabolites There is evidence of significant species-dependent differences in the degree of de-epoxidation of nivalenol in non-ruminants, and this de-epoxidation may occur in the lower gastrointestinal tract of some species. De-epoxidized metabolites were detected in the feces of mice, pigs, and laying hens, but not in mice or broilers, and based on in vitro studies, it is unlikely to form in humans. In ruminants, as with other trichothecene toxins, nivalenol may undergo extensive de-epoxidation in the rumen before absorption. Nivalenol is metabolized to de-epoxidized nivalenol. The cytotoxicity of the deepoxidized metabolites of the trichothecene toxins nivarenol (NIV) and deoxynivarenol (DON) was determined and compared with that of the corresponding toxins with intact epoxy groups and their acetylated derivatives. Cytotoxic effects were determined by a 5-bromo-2'-deoxyuridine (BrdU) incorporation assay, which assessed DNA synthesis. The toxicity of NIV and DON was expressed as the concentration at which 50% DNA synthesis was inhibited (IC50), and both showed similar toxicity within similar micromolar concentration ranges (1.19 ± 0.06 μM and 1.50 ± 0.34 μM, respectively). In the experiments, fusarenon X (4-acetyl NIV) showed similar toxicity to NIV, and 15-AcDON showed comparable toxicity to DON. 3-AcDON showed lower toxicity than DON and 15-AcDON. In the experiment, the IC50 value of deepoxidized DON was 54 times higher than that of DON, while the IC50 value of deepoxidized NIV was 55 times higher than that of NIV. These results confirm previous findings that the deepoxidation reaction is a detoxification reaction.

Interactions
This study conducted a feeding trial to determine the effects of Fusarium graminearum extract on broiler health and production performance, and the potential protective effects of natural zeolite. Fusarium graminearum extract contains novofusarene alcohol, T-2 toxin, and diacetoxyfusarene alcohol, which showed high toxicity after intraperitoneal injection in rats. One-day-old broilers were fed four diets freely for 28 days: Group I – control group; Group II – supplemented with 0.5% zeolite; Group III – supplemented with Fusarium graminearum extract; Group IV – supplemented with both 0.5% zeolite and Fusarium graminearum extract. Broilers were sacrificed after 28 days, and their relative organ weight, white blood cell count, and serum biochemical parameters were measured. No deaths were recorded during the experiment. Fusarium graminearum extract significantly reduced broiler weight gain, feed intake, feed utilization, and water consumption (p<0.05). These parameters were also decreased in Group IV, which was fed a diet supplemented with zeolite and Imperata cylindrica extract. No significant differences were observed between group II and the control group. The relative weights of liver, kidney, heart, and gizzard were significantly increased in groups III and IV (p<0.05), while only the relative weight of liver increased in group II. Application of Imperata cylindrica extract, alone or in combination with zeolite, significantly reduced white blood cell count, serum total protein, and serum albumin levels. Application of zeolite and Imperata cylindrica extract, alone or in combination, increased serum creatinine and uric acid concentrations (p<0.05). These results indicate that sublethal doses of Imperata cylindrica extract have adverse effects on the production performance and health of broilers. Adding zeolite did not alleviate these damages; in fact, for some parameters, adding zeolite exacerbated the adverse effects of Fusarium graminearum extract. Deoxynivalenol (DON) and novofusarenol (NIV) are toxic secondary trifleenene metabolites produced by Fusarium and are commonly found in grains. This study compared the toxicity of these compounds to C57BL/6 mice, examining multiple parameters including changes in plasma biochemical parameters, immune system responsiveness, and hepatic drug metabolism. Mice received oral doses of the two toxins, either alone or in combination, at 0.071 or 0.355 mg/kg body weight, three days a week for four weeks. A single administration of 0.355 mg/kg NIV altered food intake in mice, but no significant changes were detected in body weight, organ weight, or liver protein content. NIV administration also caused significant changes in plasma total carbon dioxide and uric acid concentrations. Exposure to the toxins alone led to increased plasma IgA levels, but no significant changes were observed in in vitro cytokine production in spleen cells. Hepatic ethoxyhalogen O-dealkylase, pentoxyhalogen O-depentylase, and glutathione S-transferase activities were increased, while cytochrome P4501a and P4502b subfamily expression was upregulated. The response to combined administration of DON and NIV was similar to that of each toxin alone. However, depending on the toxin dose ratio and biochemical indicators, some reactions may be additive (plasma IgA and hepatic DCNB binding) or synergistic (plasma uric acid). Deoxynivalenol (DON) is the most common trichothecene toxin in European and North American crops. DON often coexists with other trichothecene type B toxins, such as 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV), and fusarium-X (FX). Although the cytotoxicity of individual mycotoxins has been extensively studied, data on the toxicity of mycotoxin mixtures remain limited. This study aimed to evaluate the interaction of trichothecene type B toxin co-exposure on intestinal epithelial cells. Proliferating Caco-2 cells were exposed to escalating doses of trichothecene type B toxins, either alone or in binary or ternary mixtures. Mitochondrial and lysosomal functions were assessed using the MTT assay and neutral red uptake assay, respectively, to evaluate intestinal epithelial cell toxicity. All five tested mycotoxins exhibited dose-dependent effects on proliferating intestinal cells, with the toxicity increasing in the following order: 3-ADON < 15-ADON ≈ DON < NIV << FX. Binary or ternary mixtures also showed dose-dependent effects. At low concentrations (cytotoxic effects between 10% and 30-40%), mycotoxin combinations showed synergistic effects; however, the DON-NIV-FX mixture showed antagonistic effects. At high concentrations (cytotoxic effects approximately 50%), these combinations showed additive or near-additive effects. These results suggest that the co-occurrence of low doses of mycotoxins in food and diets may be more toxic than the predicted effects of using any single mycotoxin alone. This synergistic effect should be considered given the frequent co-occurrence of trichothecene toxins in diets and the concentrations of toxins consumers are exposed to. This study investigated the effects of adding moldy barley containing mycotoxins to the diets of laying hens on their production performance and health. Health indicators included different plasma parameters and liver vitamin A and E levels. Thirty laying hens were fed three different diets: one diet supplemented with 30% toxin-free barley, and the other two diets supplemented with barley from 1997 and 1998 with different degrees of mold, respectively, for a seven-week period. The moldy diets contained low to moderate concentrations of ochratoxin A, zearalenone, deoxynivalenol, and novofusarium enol. The addition of moldy barley to the diets negatively impacted feed intake, feed conversion ratio, nutrient digestibility, egg production, and egg quality. Compared to the control group, plasma alkaline phosphatase levels were elevated, and certain biochemical blood indicators (bilirubin, uric acid, chloride, protein, albumin, and vitamin A) were also elevated or altered. Although ochratoxin A contamination was relatively low, it may have been associated with reduced feed intake and contributed to some of the aforementioned effects. Higher mycotoxin contamination and an unidentified cytotoxic component in diets supplemented with barley in 1998 may also explain the more significant effects of that diet.
/Alternative and In Vitro Tests/ Nynovial fuméol (NIV) and deoxynivalenol (DON) are trichothecene fungal toxins and are considered common food contaminants. This study aimed to investigate whether the immunotoxic effects of these trichothecene toxins could be mediated by dysregulation of dendritic cell (DC) activity. This study assessed the effects of NIV and DON on LPS-induced DC maturation using mouse bone marrow-derived dendritic cells (DCs). The study found that in LPS-treated dendritic cells (DCs), exposure to NIV and DON downregulated the expression of major histocompatibility complex (MHC) molecules and helper CD11c molecules, but had no effect on the expression of the co-stimulatory molecule CD86, while also reducing nitric oxide (NO) production. Notably, NIV, but not DON, induced DC necrosis. Furthermore, cytokine profiling showed that LPS-induced IL-12 and IL-10 expression were inhibited by these two trichothecene compounds in a dose-dependent manner. On the other hand, exposure to DON and NIV directly increased the secretion of the pro-inflammatory cytokine TNF-α. Taken together, these data suggest that the immunotoxicity of NIV and DON is related to the ability of these two trichothecene compounds to interfere with the phenotypic and functional characteristics of mature dendritic cells (DCs).

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Non-human toxicity values
Oral LD50 in mice: 38.9 mg/kg
Intraperitoneal LD50 in mice: 7.4 mg/kg
Subcutaneous LD50 in mice: 7.2 mg/kg
Intravenous LD50 in mice: 7.3 mg/kg
For more complete data on non-human toxicity values of NIVALENOL (6 types), please visit the HSDB record page.

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NIVALENOLExhibits lymphotoxicity in mice after oral exposure, characterized by increased apoptosis of thymocytes and spleen cells, and damage to lymphoid tissue structures[2]

[1]. Natural Occurrence of Nivalenol, Deoxynivalenol, and Deoxynivalenol-3-Glucoside in Polish Winter Wheat. Toxins (Basel). 2018 Feb 13;10(2).

[2]. Individual and combined mycotoxins deoxynivalenol, nivalenol, and fusarenon-X induced apoptosis in lymphoid tissues of mice after oral exposure. Toxicon. 2019 Jul;165:83-94.

12,13-Epoxy-3,4,7,15-Tetrahydroxytrien-9-en-8-one has been reported in Fusarium graminearum, Fusarium culmorum, and Fusarium tricinctum, with relevant data available.

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Mechanism of Action
Trichothecene compounds, such as novofusinol, inhibit peptidyl transferases, thereby inhibiting peptide bond formation. The target organelles of trichothecene compounds are the 60S subunit of the eukaryotic ribosome, and their protein-inhibiting activity is closely related to ribosome affinity. Protein inhibition mechanisms can be divided into two types: one is the inhibition of the initial step of protein synthesis (type I), and the other is the inhibition of the elongation-termination step (type ET). Novofusinol acts on the initial step of protein synthesis, with an ID50 of 2.5 mg/mL in rabbit reticulocytes. Trichothecene toxins are potent direct and indirect inhibitors of protein, DNA, and RNA synthesis, exhibiting particularly high toxicity to tissues with high cell proliferation rates. Fusarium noctuidae rapidly inhibits protein synthesis in HELA cells and yeast spheroids. Fusarium noctuidae is a potent and highly selective inhibitor of polypeptide chain initiation in eukaryotes. Deoxynivalenol (DON) and NIV are secondary metabolites produced by trichothecene fungi. Trichothecene toxins can cause immune dysfunction, leading to a variety of responses to infection. This study evaluated the effects of DON and NIV on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264 cells. The results showed that LPS-induced NO production was reduced in the presence of these toxins. These toxins also inhibited the transcriptional activation and expression of LPS-induced inducible nitric oxide synthase (iNOS). DON or NIV suppressed LPS-induced interferon-β (IFN-β) expression, while IFN-β plays an indispensable role in LPS-induced iNOS expression. These results indicate that DON and NIV inhibit LPS-induced production of nitric oxide (NO) and IFN-β, both of which play important roles in host defense against pathogen invasion, suggesting that the inhibition of these factors may be related to the immunotoxic effects of these fungal toxins.

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Nystatin is a type B trichothecene toxin produced by Fusarium spp. [1]
; Nystatin is naturally found in Polish winter wheat, and its concentration varies depending on the growing region and environmental conditions [1]
; compared with single exposure, Nystatin combined exposure with deoxynystatin and fusarium ketone-X showed an additive or synergistic effect on inducing apoptosis in mouse lymphoid tissue cells [2]

C15H20O7

312.3151

312.12

23282-20-4

Nivalenol-13C15;911392-40-0

5284433

White to off-white solid powder

1.6±0.1 g/cm3

585.1±50.0 °C at 760 mmHg

222-223ºC

221.9±23.6 °C

0.0±3.7 mmHg at 25°C

1.658

-0.75

4

7

1

22

588

8

CC1=C[C@@H]2[C@]([C@@H](C1=O)O)([C@]3([C@@H]([C@H]([C@H]([C@@]34CO4)O2)O)O)C)CO

UKOTXHQERFPCBU-XBXCNEFVSA-N

InChI=1S/C15H20O7/c1-6-3-7-14(4-16,11(20)8(6)17)13(2)10(19)9(18)12(22-7)15(13)5-21-15/h3,7,9-12,16,18-20H,4-5H2,1-2H3/t7-,9-,10-,11-,12-,13-,14-,15+/m1/s1

(1S,2R,3S,7R,9R,10R,11S,12S)-3,10,11-trihydroxy-2-(hydroxymethyl)-1,5-dimethylspiro[8-oxatricyclo[7.2.1.02,7]dodec-5-ene-12,2'-oxirane]-4-one

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~50 mg/mL (~160.09 mM)

Solubility in Formulation 1: ≥ 5 mg/mL (16.01 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 5 mg/mL (16.01 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 5 mg/mL (16.01 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)