High Concentrations Of Zearalenone Research Articles

Exposure of Cattle Breeding Herds to Naturally Co-Contaminated Zearalenone and Deoxynivalenol: The Relevance of a Urinary Mycotoxin Monitoring System for Herd Health and Food Safety

The widespread presence of Fusarium mycotoxins in animal feed is a global issue, not only for the health of livestock but also for ensure the safety of food as an end product. High concentrations of zearalenone (ZEN) and deoxynivalenol (DON) have been detected in the diets of Japanese Black (JB) and Holstein Friesian (HF) breeding herds. Consequently, we monitored serum biochemical parameters over a long time in both herds, focusing on anti-Müllerian hormone (AMH) levels and acute-phase inflammation. Additionally, urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) and progesterone levels were measured in the HF herd. The JB herd, a ZEN-dominant model with low DON contamination, demonstrated ZEN levels that exceeded the Japanese limit in the purchased total mixed rations (TMR). Conversely, the HF herd, which primary consumes DON-dominant feed with low ZEN contamination, had high DON levels in the dent corn silage. Specifically, the JB herd’s TMR contained 1.79 mg/kg ZEN and 0.58 mg/kg DON, whereas the HF herd’s silage had 15.3 mg/kg DON (dried sample) and 0.1 mg/kg ZEN. Enzyme-linked immunoassay were used to measure urinary ZEN-DON levels following confirmation through liquid chromatography-tandem mass spectrometry. Urinary ZEN-DON levels measured were significantly correlated (p < 0.05, r > 0.6) in both herds. In the HF herd, AMH levels increased (p = 0.01) and serum amyloid A (SAA) levels decreased (p = 0.02) when contaminated and at the end of the monitoring period. Additionally, urinary ZEN and DON levels were significantly correlated with SAA levels (ZEN: p = 0.00, r = 0.46; DON: p = 0.03, r = 0.33), with an increase in ZEN and DON levels resulting in higher SAA levels. The JB herd showed no significant differences. Additionally, in the HF herd, 8-OHdG/Cre levels increased significantly during major contamination periods (p < 0.05). Clinical data from the HF herd indicated an increase in mastitis cases and treatment rates during periods of major contamination. Abortion rates in the HF herd decreased from 22.9% (before monitoring) to 8.9% (during the high contamination period) and finally to 1% (at the end of the monitoring period), with corresponding increases in progesterone levels. ZEN-DON contamination adversely affects breeding cattle’s productivity, reproductive performance, and health. Therefore, monitoring urinary ZEN-DON is valuable for detecting contaminants and ensuring the safety of food products.

Endoplasmic reticulum stress is involved in mycotoxin zearalenone induced inflammatory response, proliferation, and apoptosis in goat endometrial stromal cells

Zearalenone (ZEA) is an estrogen-like mycotoxin and is considered a secondary metabolite produced by Fusarium fungi, which are widely found in the surrounding environment. ZEA has been found to cause reproductive dysfunction in female and male animals, but the underlying mechanism remains unclear. Therefore, this study examined cell proliferation, cell apoptosis, autophagy protein expression, and some inflammatory cytokines such as IL-1β and IL-8 of goat endometrial stromal cells (ESCs) induced by different concentrations (0, 15, 30, 60, and 90 µM) of ZEA. The apoptosis rate was detected by flow cytometry. Western Blot and ELISA assay were used to identify the ER stress signaling pathway and some inflammatory cytokines. Our results revealed that ZEA induced cell proliferation and inhibited cell apoptosis at low and middle concentrations, while at high concentrations of ZEA, cell apoptosis was induced in ESCs. Additionally, ZEA induced the ER stress protein markers such as ATF6, IRE1α, EIF2α, and ATF4. LC3 as a marker of autophagy was up-regulated at all concentrations of ZEA. Moreover, IL-1β and IL-8 showed down-regulation at a low concentration of ZEA, but middle and high concentrations showed up-regulation. In the present study, Knockdown ERN1 can inhibit autophagy and the main markers of ER stress. These results suggest that the IRE1 pathway can reduce apoptosis protein markers, down activate IRE1, and unfolded protein response branches such as ATF6 and LC3 in ESCs. Additionally, IL-1β and IL-8 achieve up-regulation under knockdown IRE1, which can block ER stress markers.

Changes in antioxidant enzymes and locomotor activity of yellow mealworm larvae fed the mycotoxin zearalenone supplemented diet

The larvae of the yellow mealworm Tenebrio molitor represent a new sustainable source of protein and nutrients for animal and human nutrition. To ensure safe insect-based foods, the risks of mycotoxins and other chemicals must be elucidated. Since oxidative stress is known to underlie many mycotoxin-induced disorders, the present work investigates the effects of the mycotoxin zearalenone (ZEA) on the responses of some oxidative stress biomarkers, superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) in the whole body of mealworm, as well as the effects on larval survival, body weight, and locomotor activity, and some aspects of mycotoxin accumulation. Larvae were fed artificially contaminated wheat bran containing approximately 0.1 mg/kg, 0.5 mg/kg, 2 mg/kg, and 5 mg/kg ZEA for two weeks. The results of the current study showed that ZEA-contaminated wheat bran at a concentration of 0.5 mg/kg was correlated with increased SOD, CAT, and GST enzyme activities and decreased locomotor activity. Higher concentrations of ZEA, i.e., 2 and 5 mg/kg caused enzymes and locomotor activity to return to control levels. The presence of ZEA in the diet for two weeks did not affect survival but resulted in lower weight gain in T. molitor larvae. The concentration of ZEA in the residual material was 21.1–37.6%, while in the larvae it accounted for 0.38–0.51% of the mycotoxin in the feed. The selected concentrations were in accordance with the EU limits for the presence of ZEA in food and with the levels found in naturally contaminated cereals. Our results suggest that ZEA at the tested concentration induces physiological, biochemical, and behavioral changes in larvae that could potentially serve as early markers of mycotoxin contamination.

Zearalenone Biodegradation by the Lactobacillus Spp. and Bacillus Spp. In Vitro

Abstract In this study, the biodegradation of zearalenone (ZEN) by cell suspensions of various Lactobacillus species (Lb. fermentum 2I3 (L1), Lb. reuteri L26 (L3), Lb. plantarum L81 (L4), Lb. reuteri 2/6 (L5), Lb. plantarum CCM 1904 (L6)), Bacillus subtilis CCM 2794 (Bs), and Bacillus licheniformis CCM 2206 (Bl); was investigated in vitro. All lactobacilli cell suspensions showed very good degradation efficiency (57.9—100 %) for zearalenone at the concentration 0.01 ppm. At higher concentrations of zearalenone, their biodegradation activity decreased significantly (0—13.9 %). Bacillus subtilis CCM 2794 was able to degrade zearalenone at concentrations of: 0.01 ppm (100 %), 0.1 ppm (74.5 %), and at higher concentrations of ZEN (1 ppm; 10 ppm), the degradation was 11.7 % and 0 %, respectively. For Bacillus licheniformis CCM 2206, no biodegradation of zearalenone was observed at the concentration of 10 ppm, but slight degradation (4.5—8.8 %) was found at lower zearalenone concentrations.

Concentrations of zearalenone and its metabolites in female wild boars from woodlands and farmlands

As a valuable food base and safe shelter, large agricultural fields create favourable living conditions for wild boars for most of the year. The occurrence of mould in these fields, causing hormonal disturbances, may lead to a lower age of sexual maturity and a prolonged breeding season, and therefore, the population growth in farmlands can be markedly higher than that in extensive woodlands. This study was initiated because of reproductive cycle disturbances in wild boar populations, which were presumably linked with habitat and food types, especially rotten maize. To determine if the major sources of zearalenone (ZEN) in female wild boars are maize cobs infected with mycotoxins, we compared the concentrations of ZEN and its metabolites in organs, tissues and body fluids of wild boars hunted in extensive maize fields and in extensive woodlands where no large maize fields were located within a distance of several km. Samples of blood, bile, liver, kidneys, muscles, urine, stomach and colon contents as well as ovaries from young female wild boars (40–60 kg) were collected in 2011–2014 and prepared for liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The results showed that ZEN was present in most of the samples, from both farmlands and woodlands, but its concentration within individual types of the analysed body fluids or tissues differed depending mainly on habitat type. In nearly all the analysed cases, higher concentrations of ZEN and its derivatives were detected in the samples collected from wild boars in farmlands, suggesting that wild boars living and feeding in extensive maize fields take in markedly higher amounts of mycoestrogens.

A FUSARIOTOXINS IN FEED OF DAIRY COWS AND CARRY-OVER TO MILK AND IMPACT ON RAW MILK QUALITATIVE INDICATORS

The research aims of this study were to determine and to evaluate the effect of feed contaminated with different concentrations of fusariotoxins (zearalenone (ZEA), deoxynivalenol (DON) on the quality of milk investigate concentration zearalenone in cow milk samples. Feed and milk samples were collected from dairy farms which were the high Total Bacterial Count (TBC) and Somatic Cell Count (SCC) in milk. Feed samples were tested by TLC (Romer Labs methods). The zearalenone (ZEA) in the milk was measured by ELISA test, RIDASCREEN® Zearalenon (R-Biopharm AG). The milk qualitative indicators testing was conducted by public enterprise „Pieno tyrimai“ (Lithuania). From 12 dairy cow farms, which have high TBC and SCC, were selected feed samples for detection of fusariotoxins. Average ZEA concentration in feed was 496.25±51 µg/kg (p<0.05), DON average concentration in feed was 65.31±12 µg/kg (p<0.05). Higher amount TBC in milk samples were in those farms where was detection ZEA and DON concentration. As a result, we can assume that the higher TBC in milk samples than 221±20 thous. CFU/ml was find in farms where in feed samples was detected the higher ZEA concentration than 700±50 µg/kg (p<0.05). The highest levels than 650±42 of SCC in milk were found when the feed samples were contaminated ZEA - 700±50 µg/kg (p<0.05) and more. It can also be observed that the higher the ZEA concentration in the sample is, the higher the amount of SCC follows. Comparing concentration ZEA in feed and in milk established percent from 0.01 to 0.31 % (p<0.05). The highest levels of SCC and TBC were found in samples with high concentrations of ZEA. In 16 % milk samples we detected ZEA, concentration was from 0.08 µg/l to 0.4 µg/l. &nbsp

Zearalenone Induces Endoplasmic Reticulum Stress and Modulates the Expression of Phase I/II Enzymes in Human Liver Cells.

Zearalenone (ZEN) is a mycotoxin produced by Fusarium species; however, its mechanisms of action in human livers have not been fully elucidated. Thus, we investigated the toxic mechanisms of ZEN in human liver cells. HepG2 cells were treated with ZEN (0–40 μg/mL) for up to 24 h. A significant decrease in cell viability was observed after treatment with 20 and 40 μg/mL of ZEN, including a significant increase in apoptosis and reactive oxygen species production. ZEN increased GRP78 and CHOP, and eIF2α phosphorylation, indicating ER stress; elevated transcription of the autophagy-associated genes, beclin1 and LC3, and translation of LC3; and increased phase I metabolism by increasing PXR and CYP3A4. The protein expression level of CYP3A4 was higher with ZEN treatment up to 20 μg/mL, but remained at the control level after treatment with 40 μg/mL ZEN. In phase II metabolism, Nrf2 activation and UGT1A expression were increased with ZEN treatment up to 20 μg/mL. Treating cells with an ER stress inhibitor alleviated ZEN-induced cell death and autophagy, and inhibited the expression of phase I/II enzymes. Overall, high ZEN concentrations can modulate the expression of phase I/II enzymes via ER stress and reduced protein levels in human liver cells.

Systems to reduce mycotoxin contamination of cereals in the agricultural region of Poland and Kazakhstan

Field trials were conducted at two different geographical locations to study the effect of pesticide protection and its consequence on the mycotoxin level, ergosterol and quality parameters of wheat. The treatments involved the application of: herbicide (aryloxyalcanoic and benzoic acid), a set of two (benzimidazole and strobilurin) or three (triazole and morpholine) fungicides and a mix of herbicide and fungicides. Polish and Kazakh varieties of wheat were monitored in a three-year study. Weed populations were controlled by MCPA and dicamba, and a reduction of mycotoxin in wheat grain was observed at both geographical locations. The most significant reduction of the mycotoxin (trichothecenes, fumonisins and zearalenone) levels resulted from the application of combined MCPA/dicamba (BBCH 19–28) and thiophanate methyl/azoxytrobin (BBCH 44–58) and propioconazole/cyproconazole/tebuconazole/triadimenol/spiroxamine (BBCH 68–77). The highest concentrations of zearalenone and deoxynivalenol were detected in the control plots (571.0 and 151.0 μg/kg). The relationships between the fungal biomarker ergosterol and mycotoxins, were observed. The highest levels of ergosterol and contamination with mycotoxin were obtained for the Kazakh and Polish cereals in 2016.

Individual and combined cytotoxicity assessment of zearalenone with ochratoxin A or α-zearalenol by full factorial design.

The combined mycotoxins zearalenone (ZEA) with ochratoxin A (OTA) or α-zearalenol (α-ZOL) are frequently found together in milk. Toxicological data concerning the combined effects of these mycotoxins are sparse. In present study, individual and combined ZEA, OTA and α-ZOL caused cytotoxicity and oxidative damage, including reductions in intracellular superoxide dismutase and glutathione peroxidase activities and glutathione content, along with increases in malonaldehyde content on human Hep G2 cells after 48h of exposure. Among individual mycotoxins, OTA had the greatest cytotoxic effect followed by α-ZOL. Compared with individual mycotoxins, combinations produced more serious negative effects, more importantly, ZEA+OTA was antagonistic for these effects, whereas ZEA+α-ZOL was antagonistic at low concentrations, but synergistic at high concentrations of ZEA, which were evaluated by 3×3 full factorial analysis and estimated marginal means plots. Our results also demonstrated a significant correlation between cytotoxicity and oxidative damage in response to these combinations.

Presence of Fusarium mycotoxins in feedstuffs and cow milk sampled from Croatian farms during 2015

Mycotoxins may contaminate food of animal origin due to the carry-over effect and represent a potential risk to human health. The problem of Fusarium mycotoxin contamination becomes an issue especially during rainy years characterised by substantial temperature changes. The aim of this study was to investigate into the level of Fusarium mycotoxins zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FUM) in maize silage (n=21), concentrated dairy cattle feeds (n=56) and cow milk samples (n=105), taken during 2015 from households located in four Croatian regions. The presence of mycotoxins was determined using validated ELISA methods. A high level of feedstuffs’ contamination was evidenced, especially with ZEN, with values higher than recommended observed in 9.5 % of maize silage samples. Fourteen point three percent (14.3 %) of milk samples were DON positive, with the toxin concentrations ranging from 5.4 to 67.3 μg/L. ZEN was determined in 94.3 % of milk samples, ranging from 0.3 to 88.6 μg/L. FUM were not detected in any of the analysed milk samples. Given the tolerable daily intakes (TDIs) defined for these mycotoxins, human health risks arising from the consumption of cow milk can generally be considered low, even in times characterised by weather conditions that facilitate the production of Fusarium mycotoxins in cereals subsequently used as dairy cattle feed. The exception represents particular milk samples in which high ZEN concentrations were found.

Zearalenone and its metabolites in the tissues of female wild boars exposed per os to mycotoxins

The study was performed on 18 clinically healthy female wild boars with initial body weight of 35 ± 2 kg. The animals were divided into two experimental groups (group I and group II) and one control group (group C) of 6 female wild boars per group. Group I animals were administered per os pure zearalenone (ZEN) at 150 μg/kg BW every two months for 7 subsequent days, whereas group II animals received feed naturally contaminated with ZEN at 50 μg/kg BW/day. Female wild boars were exposed to ZEN over a period of 1 year. Control group animals were fed a placebo. Tissue samples (dorsal muscles, left lobe of liver, left kidney, spleen, apical part of the cardiac muscle, cranial lobe of lung, left ovary, central part of the left horn of the uterus) were collected on the last day of the experiment within 3 min after slaughter. In group I, the highest ZEN levels were noted in the spleen (19.813 ng/g), cardiac muscle (18.105 ng/g) and kidneys (14.555 ng/g). In group II, the highest concentrations of ZEN were observed in muscle tissue (12.033 ng/g), uterus (10.821 ng/g) and kidneys (10.463 ng/g). The highest values of the carry-over factor were noted in the same tissues. In the examined female wild boars, per os exposure to natural sources of the parent substance or a combination of ZEN and its metabolites led to different concentrations of ZEN in the analyzed tissues. Zearalenone concentrations were compatible with CF values in both experimental groups.

Effects of the Fusarium toxin zearalenone and/or deoxynivalenol on the serum IL-1, IL-4, and C3 levels in mice

ABSTRACTThe aim of this study was to evaluate the effects of the Fusarium toxin zearalenone (ZEA) and/ or deoxynivalenol (DON) on the serum IL-1, IL-4, and C3 levels in mice. In our study, 360 healthy adult female mice were randomly assigned to 9 groups for a 12-day study. Mice of all groups were given a 4-day continuous intraperitoneal injection with different concentrations of ZEA alone, DON alone, and the mixture of ZEA and DON, individually. Blood samples were collected in days 0, 3, 5, 8, and 12 of the experiment. The experiment results showed that all of the experimental groups could cause the dysregulation of the IL-1, IL-4, and C3 levels, thus affecting the immunity of mice. The mixture groups, especially in the group with higher concentrations of ZEA and DON (Group D2Z2), showed more obvious effect on the dysregulation of the IL-1, IL-4, and C3 levels. There was a synergistic effect between DON and ZEA.

In vitro zearalenone adsorption by a mixture of organic and inorganic adsorbents: application of the Box Behnken approach

Zearalenone (ZEA) adsorption by a mixture of organic (yeast cell wall) and inorganic (activated charcoal) adsorbents was evaluated by an incomplete Box Behnken (33) statistical design with a quintuplicate at the central point. The variables analysed were different ratios of adsorbents (yeast cell wall and activated charcoal) at 100:0, 87.5:12.5 and 75:25, pH (3.0, 4.5 and 6.0) and ZEA concentrations (300, 750 and 1,200 ng/ml). The adsorbent mixture at 75:25 showed higher efficiency for ZEA adsorption (≯96.1%) than the 87.5:12.5 ratio (81.3 to 93.7%) and with the pure yeast cell wall (78.1 to 55.7%). The significant variables were the ratio of adsorbent mixture and ZEA concentration. The effect of pH was not significant (P=0.05), indicating that the binding between ZEA and the adsorbent would be stable at different pH (3.0, 4.5 and 6.0). The quadratic model obtained by the Box Behnken (33) design can be used for predictive purposes, because it showed a non-significant deviation (P=49.54%) and a good correlation coefficient (R2=0.98), suggesting that the ZEA adsorption would be maximum (100%) when the adsorbent mixture is set at 75:25 and the ZEA concentration at 300 ng/ml. Although the predictive model showed that an increase in adsorption efficiency could occur in a smaller ZEA concentration (300 ng/ml), the mixture at the 75:25 ratio presented high efficiency (≯98%) in adsorption when high ZEA concentrations were used (1,200 ng/ml), indicating that these mixtures would be able to adsorb a wide range of ZEA concentrations. Therefore, this mixture of yeast cell wall and activated charcoal adsorbents at 75:25 might be a candidate for furtherin vivo testing.

Effects of the Fusarium toxin zearalenone (ZEA) and/or deoxynivalenol (DON) on the serum IgA, IgG and IgM levels in mice

The aim of this study was to evaluate the effects of the Fusarium toxin zearalenone (ZEA) and/or deoxynivalenol (DON) on the serum IgA, IgG and IgM levels in mice. In our study, 360 healthy adult female mice were randomly assigned to nine groups for a 12-day study. Mice of all groups were given a 4-day continuous intraperitoneal injection with different concentrations of ZEA alone, DON alone and the mixture of ZEA and DON, individually. Blood samples were collected on days 0, 3, 5, 8 and 12 of the experiment. The experiment results showed that all the experimental groups could cause the dysregulation of the immunoglobulin, thus affecting the humoral immune of mice. The mixture groups, especially in the group with higher concentrations of ZEA and DON (Group D2Z2), showed more obvious effect on the dysregulation of the immunoglobulin.

Zearalenone contamination in rainbow trout farms in north-eastern Poland

Zearalenone (ZEN) is a mycotoxin of potent estrogenic (hormone mimicking) activity in livestock animals, and it has frequently been found to be present in plant material worldwide, resulting in contamination of animal feed and food products. Recently, ZEN has also been reported in soil, drainage water, wastewater effluents, and rivers. However, the problem with ZEN in aquaculture remains relatively unexamined up to now. Here, we present the concentrations of ZEN in selected organs of rainbow trout (part of the dorsal white muscles with skin, the ovary, the liver and gallbladder, and the caudal part of the intestine with its content) that were purchased from three commercial fish farms in north-eastern Poland. ZEN was not detected in the trouts' muscles, and in the liver and the intestines only trace amounts of the mycotoxin were found (<2.0μg·kg−1). Interestingly, the highest concentrations of ZEN were found in the fish's ovaries (up to 7.1μg·kg−1). Additional analyses of system (surface) water and fish feed samples from the farms indicate that animal feed may be a possible source of ZEN contamination (concentration up to 81.8μg·kg−1). These results suggest that ZEN contamination may pose little health risk (if any) to the consumers of the fish. However, accumulation of this mycotoxin in the ovaries may be a concern for the aquaculture industry. Further research should evaluate the scale of this problem and answer whether the concentrations of ZEN found in feed affect fish production, especially reproduction.

Soybean isoflavone reduces the residue of zearalenone in the muscle and liver of prepubertal gilts

The aim of the present study was to evaluate the protective effects of isoflavone (ISO) against zearalenone (ZEA) residues in the muscle and liver tissues of prepubertal gilts. Seventy 75-day-old, prepubertal, female pigs (Duroc × Landrace × Yorkshire, 26.5 ± 0.60 kg) were allocated randomly to seven diet treatments for 21days as follows: one control group (fed the basal diet) and six groups fed the basal diet with the addition of either 0.5 or 2.0 mg/kg ZEA plus either 0, 300 or 600 mg/kg ISO. The results showed that the diet with 2.0 mg/kg ZEA added caused an increase of ZEA residue level in muscle tissue (P < 0.05), and that the addition of both 0.5 and 2.0 mg/kg ZEA increased the residue level of ZEA in the liver of prepubertal gilts (P < 0.05). Addition of 600 mg/kg ISO to 2.0 mg/kg ZEA-contaminated diet decreased the ZEA residue level in liver tissue (P < 0.05), and the addition of 300 or 600 mg/kg ISO to the 2.0 mg/kg ZEA-contaminated diet decreased the residue levels of ZEA in muscle tissue (P < 0.05). Western blot analysis demonstrated that feeding ZEA to prepubertal gilts increased their protein expression of 3α/3β-hydroxysteroid dehydrogenase (HSD; P < 0.05), and that the addition of 300 or 600 mg/kg ISO to the 2.0 mg/kg ZEA-contaminated diet decreased the protein expression of 3α/3β-HSD (P < 0.05), compared with the addition of 2.0 mg/kg ZEA alone. The results demonstrated that muscle and liver tissues retain residual ZEA when pigs are fed a diet contaminated with high concentrations of ZEA, and that the concentration of ZEA in muscle and liver tissues increased with increased amounts of ZEA in the feed. In diets contaminated with high levels of ZEA, the addition of ISO may accelerate the biotransformation and degradation of ZEA and its metabolites, and reduce the residues of ZEA in liver and muscle tissues of prepubertal gilts.

Concentrations of Proinflammatory Mediators of The Arachidonic Acid Cascade in Serum of Sheep with Natural Zearalenone Intoxication

Abstract The clinical form of natural zearalenone intoxication was observed in sheep that were kept indoors and fed a constant diet of feed concentrates containing high concentrations of zearalenone and its metabolite α-zearalenol. The clinical form of the disease was not noted in the control group, consisting of sheep that were kept on a pasture from spring to late autumn; only in the winter they were fed wheat pellets, in which the zearalenone concentration was determined to be the lowest among all used feed concentrates. During the course of natural zearalenone intoxication, metabolism of arachidonic acid increased, mainly due to enzymes of the cyclooxygenase group, which are responsible for the generation of prostaglandin F2α and thromboxane B2, and an increase in their concentration. Increased production of F2α and B2 was closely correlated with the serum level of SAA, an indicator of the intensity of the inflammatory reaction. This indicates that both compounds participated in the development of inflammatory reactions in the terminal end of the digestive tract that accompanied zearalenone intoxication in sheep. The imbalances noted between the eicosanoid classes investigated in this study were fundamentally responsible for the development of clinical symptoms in sick sheep that showed symptoms of partial or total prolapse of the anus and rectum and prolapse of the large intestine, which were the direct cause of the animals death.

Zearalenone risk in European wheat

Zearalenone is an oestrogenic mycotoxin produced by several Fusarium species which can infect cereals, in particular wheat and maize. The predominant species responsible for zearalenone production is Fusarium graminearum, which also produces the trichothecene mycotoxin, deoxynivalenol. The infection and subsequent mycotoxin production by Fusarium spp. is largely dependent on rainfall at flowering and before harvest. High concentrations of zearalenone in particular, appear to be produced during delayed wet harvests. There has been a recent and ongoing increase in F. graminearum incidence resulting in increased occurrence of deoxynivalenol and zearalenone across northern Europe, where delayed wet harvests are more common. Zearalenone contamination of grain is largely restricted to the outer layers of wheat grain and therefore is partitioned into the bran fraction during milling. This results in higher concentrations of zearalenone in high fibre cereal products. After the delayed wet harvest in the UK in 2008, 29% of wheat at harvest exceeded the European limit for unprocessed cereals of 100 µg/kg. This resulted in difficulties in sourcing bran which would allow production of high fibre breakfast cereals within zearalenone limits and the European Commission provided a temporary derogation for high fibre breakfast cereals of 100 µg/kg zearalenone until 31 October 2009. Rainfall data and zearalenone concentrations in UK wheat from the last ten years were used to predict the occurrence of high zearalenone in wheat (10% or more of unprocessed wheat greater than 100 µg/ kg zearalenone). High zearalenone was predicted to occur one year in five for northern Europe which matches the observed incidence for the UK within the last ten years. As a consequence, current legislative limits for zearalenone in cereals and cereal products could impact on the availability of high fibre cereal products one year in five.

A study of zearalenone cytotoxicity on human peripheral blood mononuclear cells

The mycotoxin zearalenone (ZEA) is a common contaminant of all major cereal grains worldwide with estrogenic and anabolic activity. We investigated the in vitro cytopathic effects of ZEA on freshly isolated human peripheral blood mononuclear cells (PBMC) in relation to proliferation and cell death patterns of untreated and mitogen-activated cells. The higher concentration of 30 μg/ml ZEA was found to totally inhibit T and B lymphocyte proliferation from the stimulation with phytohemagglutinin and pokeweed mitogen. The inhibitory effects of ZEA were further related to cell necrosis/apoptosis. Flow cytometry analysis showed a distinct necrotic effect on PBMC, irrespective of mitogen stimulation, whereas apoptotic activity was less evident. Necrosis was observed in both the lymphocyte and monocyte/granulocyte gates. Measurements of ZEA-induced intracellular calcium ion (Ca 2+) mobilization showed an increase of both Ca 2+ levels and the number of cells with high Ca 2+ only in the monocyte/granulocyte gated cells. Using phenylmethyl sulfonyl fluoride (PMSF), a serine protease inhibitor, and ammonium chloride (NH 4Cl), a lysosomal inhibitor, both associated with cell necrosis inhibition, we showed that PMSF at 0.05 mM and NH 4Cl at 1 and 10 mM reduced the cytopathic effects induced by 30 μg/ml ZEA, whereas apoptosis was less affected. Expose of PBMC to 1 μg/ml ZEA did not alter the viability of the cells. Our results suggest that high ZEA concentrations in the blood may well exert cytotoxic effects that merit further investigation.

Mycotoxins in Soybean Feedstuffs Used in Germany

Fifty samples of soybean meal-25 of them samples of high-protein soybean meal-and one sample of soybean hulls obtained from the feed industry in Germany were analysed for the mycotoxins aflatoxins, deoxynivalenol (DON), zearalenone (ZON) and ochratoxin A (OTA). Additionally, 4 samples of high protein soybean meal which were suspected of containing high mycotoxin levels were analysed. Aflatoxin B1 (AFB1) was detected in 32 of the 51 non-suspicious samples but the maximal concentration was only 0.41 μg/kg. DON could not be detected in any of the non-suspicious samples. ZON was detected in 23 of the 51 samples with a maximal concentration of 18 μg/kg. It was present in 18 of 25 samples of soybean meal and in the sample of soybean hulls but only in 4 of 25 samples of highprotein soybean meal. This finding suggests that ZON is mainly located in the hulls of soybeans, because high-protein meal does not contain hulls. OTA was found in 4 samples, with the greatest concentration detected being 1 μg/kg.All of the four suspicious samples of high protein soybean meal contained high ZON concentrations of up to 363 μg/kg. The contamination with other mycotoxins was on the same order of magnitude as in the case of the non-suspicious samples.