Concentrations Of Zearalenone Research Articles

Actinobacteria as Promising Biocontrol Agents for In Vitro and In Planta Degradation and Detoxification of Zearalenone

Zearalenone (ZEN) is a prevalent mycotoxin found in grains and grain-derived products, inducing adverse health effects in both animals and humans. The in-field application of microorganisms to degrade and detoxify ZEN is a promising strategy to enhance the safety of food and feed. In this study, we investigated the potential of three actinobacterial strains to degrade and detoxify ZEN in vitro and in planta on wheat ears. The residual ZEN concentration and toxicity in the samples were analysed with UHPLC-MS/MS and a bioluminescence BLYES assay, respectively. Streptomyces rimosus subsp. rimosus LMG19352 could completely degrade and detoxify 5 mg/L ZEN in LB broth within 24 h, along with significant reductions in ZEN concentration both in a minimal medium (MM) and on wheat ears. Additionally, it was the only strain that showed a significant colonisation of these ears. Rhodococcus sp. R25614 exhibited partial but significant degradation in LB broth and MM, whereas Streptomyces sp. LMG16995 degraded and detoxified ZEN in LB broth after 72 h by 39% and 33%, respectively. Although all three actinobacterial strains demonstrated the metabolic capability to degrade and detoxify ZEN in vitro, only S. rimosus subsp. rimosus LMG19352 showed promising potential to mitigate ZEN in planta. This distinction underscores the importance of incorporating in planta screening assays for assessing the potential of mycotoxin-biotransforming microorganisms as biocontrol agents.

A method for estimating the outer exposure of dairy cows to deoxynivalenol (DON) and zearalenone (ZEN) as a precondition for risk assessment based on inner exposure with toxin residue levels in blood and urine as indicators.

DON and ZEN residues in the blood and urine of dairy cows can be used to predict the outer exposure to DON and ZEN expressed per kilogram diet for a risk evaluation based on comparisons to critical dietary concentrations. This method was used to evaluate the exposure of dairy cows from 12 farms located in Brandenburg, Germany, fed rations with unknown DON and ZEN concentrations (N = 244). The corresponding diet concentrations predicted by different methods from analyzed blood and urine samples varied significantly amongst farms from 0 to 1.6mg/kg for DON and 0 to 3.0mg/kg for ZEN at a reference dry matter content of 88% but independently of lactational state (post-partum vs. early lactation). This significant variation was noticed below the critical dietary DON concentration of 5mg/kg, while the ZEN concentration in one farm exceeded the critical ZEN level of 0.5mg/kg markedly. Predicted DON concentrations of rations increased with the proportion of maize silage, while the high ZEN concentration found in one farm was most likely related to a higher proportion of sugar beet pulp supposedly highly contaminated by ZEN. Exceeding the critical dietary ZEN concentration and significant variations in DON contents below the critical level was not related to performance, reproductive performance, and health-related traits of cows. For a more consistent evaluation of possible associations between the inner exposure of cows to DON and ZEN, more frequent longitudinal observations of both mycotoxin residue levels and performance and health traits are required.

A new physical and biological strategy to reduce the content of zearalenone in infected wheat kernels: the effect of cold needle perforation, microorganisms, and purified enzyme

With the aim of reintroducing wheat grains naturally contaminated with mycotoxins into the food value chain, a decontamination strategy was developed in this study. For this purpose, in a first step, the whole wheat kernels were pre-treated using cold needle perforation. The pore size was evaluated by scanning electron microscopy and the accessibility of enzymes and microorganisms determined using fluorescent markers in the size range of enzymes (5 nm) and microorganisms (10 μm), and fluorescent microscopy. The perforated wheat grains, as well as non-perforated grains as controls, were then incubated with selected microorganisms (Bacillus megaterium Myk145 and B. licheniformis MA572) or with the enzyme ZHD518. The two bacilli strains were not able to significantly reduce the amount of zearalenone (ZEA), neither in the perforated nor in the non-perforated wheat kernels in comparison with the controls. In contrast, the enzyme ZHD518 significantly reduced the initial concentration of ZEA in the perforated and non-perforated wheat kernels in comparison with controls. Moreover, in vitro incubation of ZHD518 with ZEA showed the presence of two non-estrogenic degradation products of ZEA: hydrolysed zearalenone (HZEA) and decarboxylated hydrolysed ZEA (DHZEA). In addition, the physical pre-treatment led to a reduction in detectable mycotoxin contents in a subset of samples. Overall, this study emphasizes the promising potential of combining physical pre-treatment approaches with biological decontamination solutions in order to address the associated problem of mycotoxin contamination and food waste reduction.

Selective adsorption of zearalenone by a novel magnetic molecularly imprinted carbon nanomaterial.

In this paper, the objective is to immobilize molecularly imprinted polymers (MIPs) onto the surface of magnetic carbon nanoparticles (Fe3O4@SiO2@C) to develop an effective method for the adsorption of zearalenone (ZEN). The prepared products were characterized by FT-IR, SEM, TEM, XRD, VSM, TGA, and BET. The content of zearalenone in corn samples was monitored by HPLC. The results indicate that the particle size of magnetic molecularly imprinted polymers (MMIPs) is approximately 200nm. The adsorption mechanism of MMIPs was confirmed by static adsorption and dynamic adsorption experiments. The maximum adsorption capacity was 1.56mg/g, and the adsorption equilibrium was reached within 50min. The scatchard model showed that MMIPs had two binding sites, a high-affinity binding site and a low-affinity site. Kinetic second-order fitting indicates that MMIPs are mainly through chemisorption. In the actual sample application, the limit of detection (LOD) and limit of quantitation (LOQ) were 0.3mg/L and 0.9mg/L, respectively. The recovery of corn with the standard addition of ZEN was 73.6-88.1%, and the relative standard deviation (RSD) was 2.86-5.63%. The results demonstrated that MMIPs possess the advantages of straightforward operation, high precision, and cost-effectiveness, rendering them suitable for rapid ZEN detection.

Development of a corn flour certified reference material for the accurate determination of zearalenone.

A certified reference material (CRM, KRISS 108-01-002) for zearalenone in corn flour was developed to assure reliable and accurate measurements in testing laboratories. Commercially available corn flour underwent freeze-drying, pulverization, sieving, and homogenization. The final product was packed in amber bottles, approximately 14 g per unit, and preserved at -70 °C. 13C18-Zearalenone was used as an internal standard (IS) for the certification of zearalenone by isotope-dilution liquid chromatography-tandem mass spectrometry (ID-LC‒MS/MS) and for the analysis of α-zearalenol, β-zearalenol, and zearalanone by LC‒MS/MS. The prepared CRM was sufficiently homogeneous, as the among-unit relative standard deviation for each mycotoxin ranged from 2.2 to 5.7 %. Additionally, the stability of the mycotoxins in the CRM was evaluated under different temperature conditions and scheduled test periods, including storage at -70°C, -20°C, and 4°C and room temperature for up to 12 months, 6 months, and 1 month, respectively. The content of each target mycotoxin in the CRM remained stable throughout the monitoring period at each temperature. Zearalenone content (153.6 ± 8.0 µg/kg) was assigned as the certified value. Meanwhile, the contents of α-zearalenol (1.30 ± 0.17 µg/kg), β-zearalenol (4.75 ± 0.33 µg/kg), and zearalanone (2.09 ± 0.16 µg/kg) were provided as informative values.

Diallel Analysis of Wheat Resistance to Fusarium Head Blight and Mycotoxin Accumulation under Conditions of Artificial Inoculation and Natural Infection.

Breeding resistant wheat cultivars to Fusarium head blight (FHB), caused by Fusarium spp., is the best method for controlling the disease. The aim of this study was to estimate general combining ability (GCA) and specific combining ability (SCA) for FHB resistance in a set of eight genetically diverse winter wheat cultivars to identify potential donors of FHB resistance for crossing. FHB resistance of parents and F1 crosses produced by the half diallel scheme was evaluated under the conditions of artificial inoculation with F. graminearum and natural infection. Four FHB related traits were assessed: visual rating index (VRI), Fusarium damaged kernels (FDK), and deoxynivalenol and zearalenone content in the harvested grain samples. Significant GCA effects for FHB resistance were observed for the parental cultivars with high FHB resistance for all studied FHB resistance related traits. The significant SCA and mid-parent heterosis effects for FHB resistance were rare under both artificial inoculation and natural infection conditions and involved crosses between parents with low FHB resistance. A significant negative correlation between grain yield under natural conditions and VRI (r = -0.43) and FDK (r = -0.47) under conditions of artificial inoculation was observed in the set of the studied F1 crosses. Some crosses showed high yield and high FHB resistance, indicating that breeding of FHB resistant genotypes could be performed without yield penalty. These crosses involved resistant cultivars with significant GCA effects for FHB resistance indicating that that they could be used as good donors of FHB resistance.

Improve the accuracy of FT-NIR for determination of zearalenone content in wheat by using the characteristic wavelength optimization algorithm

The research contributes a unique method to achieve high-precision quantification of zearalenone (ZEN) in wheat, significantly improving accuracy in the analysis. Fourier transform near infrared spectroscopy (FT-NIR) was employed to capture the spectral information of wheat with different mildew degrees. Three feature selection models, competitive adaptive reweighted sampling (CARS), support vector machine-recursive feature elimination (SVM-RFE), and multiple feature-spaces ensemble-least absolute shrinkage and selection operator (MFE-LASSO) were introduced to processed pre-processed near-infrared spectral data and established partial least squares (PLS) regression according to the selected features. The outcomes indicated that the optimal generalization performance was achieved by the PLS model optimized through the MFE-LASSO model. The root mean square error of prediction (RMSEP) was 18.6442 μg·kg-1, coefficient of predictive determination (RP2) was 0.9545, and relative percent deviation (RPD) was 4.3198. According to the results, it is feasible to construct a stoichiometric model for the quantitative determination of ZEN in wheat by using FT-NIR combined with feature selection algorithm, and this method can also be extended to the detection of various molds in other cereals in the future.

Forsythoside A protects against Zearalenone-induced cell damage in chicken embryonic fibroblasts via mitigation of endoplasmic reticulum stress.

Zearalenone (ZEA) is a non-steroidal estrogenic mycotoxin that exerts its toxic effects through various damage mechanisms such as oxidative stress, endoplasmic reticulum stress (ERS), mitochondrial damage, cell cycle arrest, and apoptosis. At present, there are few studies on drugs that can rescue ZEA-induced chicken embryonic fibroblasts damage. Forsythoside A (FA) is one of effective ingredients of traditional Chinese medicine that plays a role in various biological functions, but its antitoxin research has not been investigated so far. In this study, in vitro experiments were carried out. Chicken embryo fibroblast (DF-1) cells was used as the research object to select the appropriate treatment concentration of ZEA and examined reactive oxygen species (ROS), mitochondrial membrane potential, ERS and apoptosis to investigate the effects and mechanisms of FA in alleviating ZEA-induced cytotoxicity in DF-1 cells. Our results showed that ZEA induced ERS and activated the unfolded protein response (UPR) leading to apoptosis, an apoptotic pathway characterized by overproduction of Lactate dehydrogenase (LDH), Caspase-3, and ROS and loss of mitochondrial membrane potential. We also demonstrated that FA help to prevent ERS and attenuated ZEA-induced apoptosis in DF-1 cells by reducing the level of ROS, downregulating GRP78, PERK, ATF4, ATF6, JNK, IRE1, ASK1, CHOP, BAX expression, and up-regulating Bcl-2 expression. Our results provide a basis for an in-depth study of the mechanism of toxic effects of ZEA on chicken cells and the means of detoxification, which has implications for the treatment of relevant avian diseases.

Determination of zearalenone content in wheat by modified one-dimensional convolutional neural network-based Raman spectra

In this study, a Raman spectral model trained based on Transformer-improved one-dimensional convolutional neural network (1D-CNN) was proposed for the quantitative analysis of zearalenone in wheat. Raman spectra of wheat flour samples with varying levels of mold contamination were acquired using a portable Raman spectrometer. The 1D-CNN was introduced to extract local features of Raman spectra using three convolutional layers, and then the Transformer module was employed to encode and decode these local features for obtaining global information. This approach ensured that the network could better learn complex input spectra data, thereby improving the detection accuracy and precision of the model. The results showed that the improved 1D-CNN model had better generalization performance compared to the 1D-CNN model, with a root mean square error of prediction (RMSEP) of 2.5759 μg/kg, a coefficient of determination (RP2) of 0.9837, and a relative prediction deviation (RPD) of 7.9516. The study demonstrated that the 1D-CNN model improved by Transformer could achieve better feature self-learning and multivariate model correction of Raman spectroscopy data. This research introduces a novel reference method for chemometric analysis of Raman spectra.

Heme Oxygenase-1 Regulates Zearalenone-Induced Oxidative Stress and Apoptosis in Sheep Follicular Granulosa Cells.

Zearalenone (ZEA) is a common non-steroidal estrogenic mycotoxin found in a range of animal feeds and poses a serious threat to the reproductive health of farm animals and humans. However, the mechanism underlying ZEA-induced reproductive toxicity in sheep remains unknown. Granulosa cells are crucial for egg maturation and the fertility of female sheep. In this study, we aimed to examine the impact of different ZEA concentrations on sheep follicular granulosa cells and to elucidate the potential molecular mechanism underlying ZEA-induced toxicity using transcriptome sequencing and molecular biological approaches. Treating primary sheep follicular granulosa cells with different concentrations of ZEA promoted the overproduction of reactive oxygen species (ROS), increased lipid peroxidation products, led to cellular oxidative stress, decreased antioxidant enzyme activities, and induced cell apoptosis. Using transcriptome approaches, 1395 differentially expressed genes were obtained from sheep follicular granulosa cells cultured in vitro after ZEA treatment. Among them, heme oxygenase-1 (HMOX1) was involved in 11 biological processes. The protein interaction network indicated interactions between HMOX1 and oxidative and apoptotic proteins. In addition, N-acetylcysteine pretreatment effectively reduced the ZEA-induced increase in the expression of HMOX1 and Caspase3 by eliminating ROS. Hence, we suggest that HMOX1 is a key differential gene involved in the regulation of ZEA-induced oxidative stress and apoptosis in follicular granulosa cells. These findings provide novel insights into the prevention and control of mycotoxins in livestock.

Effect of DON and ZEN and their metabolites DOM-1 and HZEN on B cell proliferation and antibody production.

The mycotoxins deoxynivalenol (DON) and zearalenone (ZEN), produced by Fusarium fungi, are frequently found in the cereal-rich diet of pigs and can modulate the immune system. Some enzymes or bacteria present in the digestive tract can de-epoxydize DON to deepoxy-deoxynivalenol (DOM-1) and biotransform ZEN into hydrolyzed ZEN (HZEN). The effects of these metabolites on immune cells, particularly with respect to the vaccine responses, are poorly documented. The aim of this study was to address the impact of DON and ZEN and their respective derivatives, on proliferation, and antibody production of porcine B cells in vitro. Peripheral blood mononuclear cells (PBMCs), isolated from healthy pigs, were stimulated with the Toll-like receptor (TLR) 7/8-agonist Resiquimod (R848) or the TLR/1/2-agonist Pam3Cys-SKKKK in combination with DON [0.1-1.6 µM] or DOM-1 [1.6 µM and 16 µM] and ZEN [2.5-40 µM] or HZEN [40 µM]. A strong decrease in B-cell proliferation was observed at DON concentrations equal to or exceeding 0.8 µM and at ZEN concentrations equal to or exceeding 20 µM. Treatment with 1.6 µM DON or 40 µM ZEN led to almost a complete loss of live CD79α+ B cells. Moreover, CD21 expression of proliferating IgG+ and IgM+ B-cell subsets was decreased at DON concentrations equal to and exceeding 0.4 µM and at ZEN concentrations equal to or exceeding 10 µM. ELISpot assays revealed a decrease of IgG-secreting B cells at concentrations of and exceeding 0.4 µM and at ZEN concentrations equal to and exceeding 10 µM. ELISA assays showed a decrease of IgM, IgG, and IgA secretion at concentrations equal to or exceeding 0.4 µM DON. ZEN reduced IgM secretion at 20-40 µM (both R848 and Pam3Cys-SKKKK), IgG secretion at 40 µM (both R848 and Pam3Cys-SKKKK) and IgA secretion at 20-40 µM. Our in vitro experiments show that while DON and ZEN impair immunoglobulin production and B-cell proliferation, this effect is abrogated by HZEN and DOM-1.

Adsorption of Zearalenone by Aureobasidium pullulans Autolyzed Biomass Preparation and Its Detoxification Properties in Cultures of Saccharomyces cerevisiae Yeast.

Different preventive strategies are needed to minimize the intake risks of mycotoxins, including zearalenone (ZEN). The aim of this study was to determine the ZEN adsorption ability of an autolyzed biomass preparation of polymorphic yeast Aureobasidium pullulans A.p.-3. The evaluation of the antitoxic properties of the preparation was also performed in relation to Saccharomyces cerevisiae yeast (ATCC 2366, ATCC 7090 and ATCC 9763) used as a model cell exposed to a toxic ZEN dose. The preparation at a dose of 5 mg/mL showed the adsorption of ZEN present in model systems at concentrations between 1 μg/mL to 100 μg/mL. The highest degree of adsorption was established for ZEN concentrations of 1 μg/mL and 5 μg/mL, becoming limited at higher doses of the toxin. Based on the Langmuir model of adsorption isotherms, the predicted maximum ZEN adsorption was approx. 190 µg/mL, regardless of pH. The growth of three strains of S. cerevisiae yeast cells in the medium with ZEN at concentrations within the range of 1.56 μg/mL-100 μg/mL was analyzed to determine the minimum inhibitory concentration. The growth of all tested strains was especially limited by high doses of ZEN, i.e., 50 and 100 μg/mL. The protective effect of the tested preparation was noted in relation to yeast cells exposed to toxic 100 μg/mL ZEN doses. The highest yeast cell growth (app. 36% percentage) was noted for a S. cerevisiae ATCC 9763 strain compared to the medium with ZEN but without preparation. More detailed tests determining the antitoxic mechanisms of the A. pullulans preparation are planned in the future, including cell culture bioassays and animal digestive tract models.

Toxicokinetics of Zearalenone following Oral Administration in Female Dezhou Donkeys.

Zearalenone (ZEN) is a mycotoxin produced by various Fusarium strains, that is present in food and feed raw materials worldwide, causing toxicity effects in animals and humans. This research aimed to explore the toxicokinetics of ZEN on female Dezhou donkeys following a single oral exposure dosage of 2 mg/kg BW (body weight). The sample collection of donkeys plasma was carried out at 0, 5, 10, 15, 20, 30, 45, 60, 90 min, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 6 h, 9 h, 12 h, 24 h, 48 h, 72 h, 96 h and 120 h via intravenous catheter, and fecal and urinary samples were severally collected at 0 h and every 6 h until 120 h. The concentrations of ZEN, α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), α-zearalanol (α-ZAL), β-zearalanol (β-ZAL), zearalanone (ZAN) in plasma, urine, and feces were detected by UPLC-MS/MS. Only ZEN was detected in plasma, and the maximum was 15.34 ± 5.12 µg/L occurred at 0.48 h after gavage. The total plasma clearance (Cl) of ZEN was 95.20 ± 8.01 L·kg·BW-1·h-1. In addition, the volume of distribution (Vd) was up to 216.17 ± 58.71 L/kg. The percentage of total ZEN (ZEN plus the main metabolites) excretion in feces and urine was 2.49% and 2.10%, respectively. In summary, ZEN was fast absorbed and relatively slowly excreted in female donkeys during 120 h after a single gavage, indicating a trend of wider tissue distribution and longer tissue persistence.

Safety of Chinese Cabbage Waste and Rice Straw Mixed Silage and Its Effect on Growth and Health Performance of Hu Sheep

Improper disposal of vegetable waste can cause serious environmental pollution, but because they contain huge water content and organic matter, they are not suitable for disposal by methods such as incineration and landfill. However, vegetable waste contains a large amount of nutrients and have some complementary effects with rice straw in terms of physical structure, nutrients, and moisture. In this experiment, the plant feed (corn husk, peanut shells and sorghum shells) was used as the control group (CON group), and the mixed silage of Chinese cabbage waste and rice straw (mixed silage) was used as the experiment group (TRE group), and its safety performance was evaluated by testing its toxin content, pesticide residues, vitamin contents and feeding experiment of Hu sheep. In the animal experiment, 16 healthy Hu sheep (5.5 months, 39.11 ± 4.16 kg) were randomly divided into two groups of 8 each. The results of the safety performance evaluation showed that the content of mycotoxins, heavy metals, and nitrites as well as pesticide residues in the crude feeds of both groups were within the range of Chinese feed hygiene standards. In addition, the levels of deoxynivalenol (DON) and aflatoxin (AFT) in the CON group were lower, while the content of ochratoxin (OTA) and zearalenone were higher than those in the TRE group (p < 0.05). The levels of plumbum(Pb), chromium (Cr), cadmium (Cd), and nitrite in the CON group were lower than the mixed silage, while the levels of As were higher than the mixed silage (p < 0.05). It is worth noticing that the content of vitamin B2 (VB2) and vitamin C (VC) in the TRE group was higher than the CON group (p < 0.05). The results of the feeding experiment showed that the mixed silage did not affect the growth performance, nutrient digestibility, organ index, and intestinal index of Hu sheep (p > 0.05). In addition, the mixed silage reduced the weight of omasum, the proportion of omasum to live weight before slaughter, the amount of compound stomach, and the proportion of compound stomach to live weight before slaughter, which were higher than those in the TRE group (p < 0.05). The thickness of the basal layer of the rumen abdominal sac, the red blood cell count, the content of IL-10, and TNF-α in the blood, and TNF-α content in the rumen of the Hu sheep in the TRE group were higher than the CON group (p < 0.05). In conclusion, the feed safety index content of the mixed silage did not exceed the Chinese feed hygiene and safety standards and did not cause adverse effects on the growth performance of the Hu sheep, and it improved the immune performance of the body and digestive tract of the sheep to a certain extent and promoted the healthy development of the sheep.

A Cohort Study Investigating Zearalenone Concentrations and Selected Steroid Levels in Patients with Sigmoid Colorectal Cancer or Colorectal Cancer.

In this study was to determine if sigmoid colorectal cancer (SCC) and colorectal cancer (CRC) in women (W) and men (M) is accompanied by zearalenone (ZEN) mycotoxicosis and changes in selected steroid levels. This cohort study was conducted on female and male patients selected from a population based on the presence of SCC or CRC, which was accompanied by the presence or absence (control group) of ZEN in their blood. The control group consisted of 17 patients with symptoms of SCC and CRC, where ZEN and its metabolites were not detected in the peripheral blood. The experimental groups comprised a total of 16 patients with SCC and CRC, where ZEN, but not its metabolites, was detected in their peripheral blood samples. In groups SCC and CRC, the ZEN levels were very high, in the range from 214 to 289 ng/mL of blood. Considerable variations were observed in the concentrations of steroid hormones. Estradiol (E2) levels ranged from 166.25 (group C) to 325 pg/mL (group CRC) in women and from 98 (group C) to 95.5 pg/mL (group CRC) in men. Progesterone (P4) levels ranged from 12.09 (group C) to 13.64 ng/mL (group SCC) in women and from 6.98 (group CRC) to 12.01 ng/mL (group C) in men. These results indicate that post-menopausal women and similarly aged elderly men have a high and individualized demand for estrogen that is relatively effectively met by ZEN, which triggers qualitative changes in estrogen receptors. The shortage of ZEN metabolites (values under the sensitivity of the method) confirmed the high estrogen demand in the studied subjects. The presence or absence of ZEN could have influenced the therapeutic outcomes in the analyzed patients.

Assessment of developmental toxicity and the potential mode of action underlying single and binary exposure to estrogenic endocrine disrupting chemicals in zebrafish (Danio rerio)

The current study investigated the effect of single and binary exposure to distinct xenoestrogens, including diethylstilbestrol (DES) and zearalenone (ZEN), on zebrafish embryos subjected to continuous exposure for 4 days starting from 4 h post fertilization. Noteworthy impact on cumulative mortality, hatchability, spinal and tail curvature, pericardial edema, and reduction in blood circulation were observed in DES-treated embryos, with lower incidence and intensity shown for ZEN at the same nominal concentration (3 μM). An interactive effect was seen for the combined exposure to DES and ZEN, in which deformities and circulatory failure mediated by DES were mitigated by co-treatment with low concentrations of ZEN. Similarly, ZEN-induced spinal and tail curvature, pericardial edema, and blood flow reduction declined dramatically following DES co-exposure at low concentrations. A significant counteracting effect has been observed against DES- and ZEN-induced developmental anomalies following co-treatment with an estrogen receptor (ER) antagonist, fulvestrant (FUL). The assessment of the aromatase gene (CYP19A1b) showed that DES strongly upregulated mRNA expression of CYP19A1b with a lower EC50 (1.1 × 10−3 nM) than a natural estrogen, 17β-estradiol (2.5 nM). Similarly, ZEN induced CYP19A1b mRNA expression with an EC50 of 57 nM. Exposure to 10 or 20 μM FUL inhibited the expression of CYP19A1b induced by a single treatment of DES or ZEN. Overall, the competitive action against ER could be the main mechanism underlying the developmental toxicity induced by DES and ZEN.

Dietary aflatoxin B1 and zearalenone contamination affected growth performance, intestinal and hepatopancreas gene expression profiles and histology of the intestine and gill in goldfish, Carassius auratus

The effects of dietary aflatoxin B1 (AFB1) and/or zearalenone (ZEA) contamination on goldfish, was investigated. Total of 540 fingerlings, 6.25 ± 0.12 g, were randomly allocated to experimental groups comprised of factorial designation of different dietary AFB1 (0, 50 and 100 ppb) and ZEA (0, 500 and 1000 ppb) concentrations for 60 days. Each treatment was conducted in triplicate. Results showed dietary AFB1 and/or ZEA depressed the growth performance of fish; the highest and lowest weight gain (WG), final body weight, thermal growth coefficient (TGC) and daily growth coefficient (DGC) were observed in control group and AFB50ZEA1000, respectively (P < 0.05). Moreover, dietary AFB1 and/or ZEA significantly affected the gene expressions of Caspase 3, Cytochrome P450 (CYP) 1A1, Glutathione S-transferases (GST) and Heat shock proteins 70 (HSP70) in a dose-dependent and tissue specific manner (hepatopancreas or intestine) (P < 0.05). In the hepatopancreas, the highest relative expression of Caspase 3 (2.08 ± 0.43), CYP1A1 (2.73 ± 1.18), GST (32.86 ± 4.03) and HSP70 (7.55 ± 2.35) genes were observed in AFB100ZEA1000, AFB50ZEA0, AFB0ZEA1000 and AFB0ZEA1000 groups, respectively (P < 0.05). However, the highest relative expression of Caspase 3 (39.70 ± 10.98), CYP1A1 (966.4 ± 68.6), GST (175.8 ± 37.1) and HSP70 (1.01 ± 0.22) genes in the intestinal tissue were observed in AFB100ZEA1000, AFB0ZEA1000, AFB100ZEA500 AFB0ZEA0 groups (P < 0.05). The highest scores of the intestine tissue inflammation (4.0 ± 0.75) and necrosis (4.0 ± 0.75) were observed in AFB100ZEA500 group (P < 0.05). The results indicated the significant contribution of the intestine in metabolism/excretion of the feed-born toxins. Furthermore, the results revealed that dietary AFB1 and/or ZEA contamination could negatively affect fish performance and detoxifying genes expression profiles at concentrations even lower than those indicated in the literature as the safe feed mycotoxin thresholds.

Preparation of Anti-Zearalenone IgY and Development of an Indirect Competitive ELISA Method for the Measurement of Zearalenone in Post-Fermented Tea.

Post-fermented tea (PFT) is one of the most commonly consumed beverages worldwide. Rapid microbial growth and significant changes in the microbial composition of PFT during processing and storage pose a potential risk of contamination with mycotoxins such as zearalenone (ZEN). Screening for ZEN contamination in a simple, rapid, and inexpensive manner is required to ensure that PFT is safe for consumption. To monitor ZEN in PFT, ZEN was conjugated with bovine serum albumin to prepare egg yolk immunoglobulins (IgY). A specific indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on IgY was developed and validated. ZEN was extracted with acetonitrile and water (50:50, v/v) containing 5% acetic acid and purified using a mixture of primary and secondary amines and graphitized carbon black to remove matrix interference from the PFT samples. Under optimal conditions, the linear range of this assay was 13.8-508.9 ng mL-1, the limit of detection was 9.3 ng mL-1, and the half-maximal inhibitory concentration was 83.8 ng mL-1. Cross-reactivity was negligible, and the assay was specific for ZEN-related molecules. The recovery rate of ZEN in the control blanks of PFT samples spiked with a defined concentration of ZEN of 89.5% to 98.0%. The recovery and accuracy of the method were qualified for PFT matrices. No significant differences were evident between the results of the actual PFT samples analyzed by high-performance liquid chromatography and ic-ELISA. The collective data indicate that the developed ic-ELISA can be used for the rapid and simple detection of ZEN in PFT products.

Effect of Mycotoxins in Silage on Biogas Production.

Mycotoxins can pose a threat to biogas production as they can contaminate the feedstock used in biogas production, such as agricultural crops and other organic materials. This research study evaluated the contents of deoxynivalenol (DON), zearalenone (ZEA), fumonisin (FUM), and aflatoxin (AFL) mycotoxins in maize silage prior to it being processed in a biogas plant and in digestate produced at the end of the anaerobic digestion (AD) process. In the experiment, three samples of silage were collected from one silage warehouse: Variant 1 = low contamination, Variant 2 = medium contamination, and Variant 3 = heavy contamination, which were subjected to investigation. A significantly reduced biogas production was recorded that was proportional to the increasing contamination with molds, which was primarily due to the AD of silage caused by technologically erroneous silage treatment. The AD was connected with changes in silage composition expressed by the values of VS content, sugar content, lactic acid content, acetic acid content, and the ratio of lactic acid content to acetic acid content. The production of biogas and methane decreased with the increasing contents of NDF, ADF, CF, and lignin. The only exception was Variant 2, in which the content of ADF, CF, and lignin was lower (by 8-11%) than that in Variant 1, and only the content of NDF was higher (by 9%) than that in Variant 1. A secondary factor that also correlated with changes in the composition of the substrate was the development of undesirable organisms, which further contributed to its degradation and to the production of mycotoxins. It was also demonstrated in this study that during the AD process, the tested mycotoxins were degraded, and their content was reduced by 27-100%. Only the variant with low mold contamination showed a DON concentration increase of 27.8%.

Quercetin Attenuates the Combined Effects of Zearalenone and Lipopolysaccharide on IPEC-J2 Cell Injury through Activating the Nrf2 Signaling Pathway.

Zearalenone (ZEA) is a mycotoxin with an estrogen-like effect that is widely found in feed. Lipopolysaccharides (LPS) derived from Gram-negative bacteria are a common endotoxin, and both toxins have effects on human and livestock health. During animal feeding, ZEA as an exotoxin and LPS as an endotoxin have the potential to co-exist in organisms. At present, other studies have only focused on the hazards of single toxins, but there are fewer studies on the coexistence and interaction between ZEA and LPS. Therefore, a further study to investigate the combined toxic effects of different concentrations of ZEA and LPS is warranted. Quercetin (QUE) is a natural flavonoid compound with strong antioxidant and anti-inflammatory properties. It is unclear whether QUE can mitigate the combined effects of ZEA and LPS. IPEC-J2, isolated from the jejunum of non-breastfed neonatal piglets, is an ideal model for the study of epithelial cell transport, intestinal bacterial interactions, and the nutrient modulation of intestinal function. Therefore, the purpose of the present study was to demonstrate the effect of QUE in alleviating the combined toxic effect of ZEA and LPS on IPEC-J2 cell damage. Cell viability was measured after treating IPEC-J2 cells sequentially with 10, 20, 30, 40, 60, 80, and 100 μM ZEA, 1, 10, 50, and 100 μg/mL LPS, and 20, 40, 60, 80, 100, and 200 μM QUE for 24 h. Based on the cell viability results, 20 μM ZEA and 1 μg/mL LPS were selected as the most suitable concentrations for further analysis. For QUE, 20 μM increased the cell viability, while 40-200 μM QUE decreased the cell viability. Therefore, for the subsequent study, 20 μM QUE was selected in combination with 20 μM ZEA and 1 μg/mL LPS. The results showed that QUE increased the cellular viability and decreased the LDH content more compared to the effects of the ZEA+LPS group. At the gene level, QUE addition up-regulated the expression of Nrf2, HO-1, SOD2, and NQO1 at the gene or protein level compared to those of the ZEA+LPS group. The measurement of tight junction-related genes and proteins showed QUE up-regulated the expression of Claudin, ZO-1, and Occludin genes and proteins more than in the ZEA+LPS group. QUE addition reduced the rate of apoptosis more than that in the ZEA+LPS group. The expressions of Bcl-2 and Bax were examined at the gene level, and QUE addition significantly reduced the Bax gene expression level compared to that of the ZEA+LPS group, but there was no apparent variation in the expression level of Bcl-2. In summary, QUE can alleviate the combined toxic effects of ZEA and LPS on IPEC-J2 cells via modulating the Nrf2 signaling pathway, up-regulating the expression of antioxidative genes, and enhancing the intestinal barrier.