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Development and Application of an Immunoaffinity Column Enzyme Immunoassay for Mycotoxin Zearalenone in Complicated Samples  [PDF]
Xiaoqian Tang, Xin Li, Peiwu Li, Qi Zhang, Ran Li, Wen Zhang, Xiaoxia Ding, Jiawen Lei, Zhaowei Zhang
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0085606
Abstract: The zearalenone (ZEA) monoclonal antibody (mAb) 2D3, one of the highest sensitivity antibodies, was developed. Based on this mAb, it was established of an immunoaffinity column (IAC) coupled with an indirect competitive enzyme-linked immunosorbent assay (icELISA). After optimization, the icELISA allowed an IC50 against ZEA of 0.02 μg L?1. The mAb 2D3 exhibited a high recognition of ZEA (100%) and β-zearalenol (β-ZOL, 88.2%). Its cross-reactivity with α-zearalenol (α-ZOL) and β-zearalanol (β-ZAL) were found to be 4.4% and 4.6%, respectively. The IAC-icELISA method was employed to analyze ZEA contamination in food samples, compared with high-performance liquid chromatography (HPLC). The spiked assay for ZEA demonstrated the considerable recoveries for IAC-icELISA (83–93%) and HPLC (94–108%) methods. Results showed that the mAb 2D3 and IAC-icELISA method posed potential applications in sensitively determination of ZEA in maize.
Effects of mycotoxin T-2 and zearalenone on histopathological changes in black tiger shrimp (Penaeus monodon Fabricius)
Bundit, O.,Kanghae, H.,Phromkunthong, W.,Supamattaya, K.
Songklanakarin Journal of Science and Technology , 2006,
Abstract: The effects of T-2 toxin and zearalenone were studied in black tiger shrimp (Penaeus monodon Fabricius). In the experiment, black tiger shrimp were fed with different concentrations of T-2 toxin, i.e. 0, 0.1, 1.0 and 2.0 ppm and zearalenone, i.e. 0, 0.1. 0.5 and 1.0 ppm. Shrimp with initial average weight of 4.7 g were experimented for a-10-wk period. Supplementation of 0, 0.1 and 1.0 ppm T-2 produced no histological changes in hepatopancreatic, hemopoietic tissue or lymphoid cell while at higher concentration of 2.0 ppm atrophy, severe necrosis and degeneration of hepatopancreatic tubules, loose contact of hemopoietic tissue and lymphoid organ occurred. Similar observations were noted for the treatments with 0.5 and 1.0 ppm zearalenone - supplemented feed. Histological changes were, however, observed in hepatopancreatic tissue. The scale of histological changes correlated with feeding period and concentrations of zearalenone shrimp received.
Influence of mycotoxin zearalenone and its derivatives (alpha and beta zearalenol) on apoptosis and proliferation of cultured granulosa cells from equine ovaries
Fiorenza Minervini, Alessandra Giannoccaro, Francesca Fornelli, Maria Dell'Aquila, Paolo Minoia, Angelo Visconti
Reproductive Biology and Endocrinology , 2006, DOI: 10.1186/1477-7827-4-62
Abstract: The cell proliferation was evaluated by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test after 3 days exposure at different concentrations of ZEA and its derivatives (from 1 × 10-7 to 0.1 microM). The apoptosis induction was evaluated after 1 day exposure, by DNA analysis using flow cytometry.An increase in cell proliferation with respect to the control was observed in the presence of ZEA at 1 × 10-3 and 1 × 10-4 microM and apoptosis was induced by all mycotoxins at different concentrations.The simultaneous presence of apoptosis and proliferation in GC cultures treated with zearalenones could indicate that these mycotoxins could be effective in inducing follicular atresia. These effects of zearalenones may result from both direct interaction with oestrogen-receptors as well as interaction with the enzymes 3alpha (beta)-hydroxysteroid dehydrogenase (HSD), involved in the synthesis and metabolism of endogenous steroid hormones. These cellular disturbances, described for the first time in equine GCs cultured in vitro, could be hypothesized as referred to reproductive failures of unknown ethiology in the mare.Many different mycotoxins have been recognized and isolated from a variety of Fusarium moulds and some of the disease states, caused by consumption of cereals containing these toxins in domestic animals as well as in humans, have been called fusariotoxicoses. Zearalenone (ZEA) and related compounds α and β zearalenol (α and β-ZOL) and α and β zearalanol (α and β-ZAL) are synthesized by a number of species of Fusarium such as F. graminearum, F. tricinctum, F. moniliforme and F. oxysporum [1].Sensitivity to the effects of mycotoxins is related to species-dependent biotransformation pathways. Zearalenone is metabolized via two pathways in hepatocytes and intestinal cells, namely conjugation with glucuronic acid and reduction to α and β-ZOL by 3 α(β)-hydroxysteroid dehydrogenase (HSD) [2-4]. These reactions show similarities to processe
Zearalenone and Reproductive Function in Farm Animals  [PDF]
Fiorenza Minervini,Maria Elena Dell’Aquila
International Journal of Molecular Sciences , 2008, DOI: 10.3390/ijms9122570
Abstract: Farm animals are exposed to zearalenone through the feed because of the widespread occurrence of this mycotoxin in cereals and clinical reproductive disorders due to mycotoxin effects are often reported in farm animal species. This review describes the metabolism, the mechanistic aspects, the clinical reproductive symptoms and the in vitro effects on functional parameters of oocytes and sperm cells induced by zearalenone and its derivatives in farm animals. The studies on in vitro effects allow to understand the action mechanisms of mycotoxins and, sometime, to explain the in vivo symptoms. The impairment of semen quality and female reproductive function induced by zearalenone could be a factor responsible for the reproductive failure in farm animals.
Occurrence of Aflatoxin B1, T-2 Toxin and Zearalenone in Compound Animal Feed
Abdurrahman Aksoy,Oguzhan Yavuz,Y. Kursad Das,Dilek Guvenc,O. Hakan Muglali
Journal of Animal and Veterinary Advances , 2012,
Abstract: This study was performed to determine mycotoxin (aflatoxin B1 (AFB1), T-2 toxin and Zearalenone (ZEA)) levels in compound ruminant feeds collected from local feed sellers in Samsun province, Turkey. Forty compound feed samples were collected and analyzed by the ELISA method. The mean and standard error of the AFB1 were 6.81±0.81 (range 0.15-32) μg kg-1. T-2 toxin and ZEA levels were 33.87±5.80 (2.24-132.2) μg kg-1 and 175.26±43.05 (51.61-1023.25) μg kg-1, respectively. The incidence of Aflatoxin B1 (AFB1), T-2 toxin and Zearalenone (ZEA) in compound feeds was found to be 95, 65 and 87.5%, respectively. Although, most of samples were found to be contaminated with mycotoxins, the levels of contamination for AFB1 and T-2 toxins were found to be relatively lower than that of ZEA levels in the feed samples. The types and levels of mycotoxins present varied. The most common contaminants were the AFB1 and ZEA. Twenty-three of the samples contained three types of mycotoxins (57.5%). Only one sample did not contain any mycotoxin (2.5%). Two samples contained only AFB1 (5%) and the rest (14 samples) contained two types of mycotoxins (35%). The mycotoxin found in highest levels was ZEA and the mycotoxin with the higher frequency of occurrence was AFB1. Correlation between the occurrence of the Fusarium toxins (T-2 toxins and ZEA) was found to be statistically significant (p<0.05).
Investigation of the Biochemical and Histological Changes Induced by Zearalenone Mycotoxin on Liver in Male Mice and the Protective Role of Crude Venom Extracted from Jellyfish Cassiopea Andromeda  [PDF]
Madeha Al-Seeni, Nagwa El-Sawi, Soad Shaker, Asma Al-Amoudi
Food and Nutrition Sciences (FNS) , 2011, DOI: 10.4236/fns.2011.24045
Abstract: Zearalenone (ZEN) is a non steroidal estrogenic mycotoxin produced by Fusarium species of fungi which contaminate human foods and animal feeds worldwide. In this study hepatotoxicity of ZEN was evaluated in mice by oral adminis-tration of single and repeated doses of ZEN mycotoxin (2.7 mg/kg b.w.). The protective effect of crude venom extracted from jellyfish Cassiopea andromeda was also assessed. Mice were divided into four groups (N = 10). G1: receiving the toxin once and sacrificed 48 h later, G2: toxin administered twice for one week, G3: toxin administered twice a week for two weeks, G4: pretreated orally by a single dose of crude venom (1.78 mg/20g) 24 hours prior to administration of ZEN twice a week for two weeks. Each treated group had its corresponding control which received 1% DMSO sa-line.ZEN treatment significantly increased alanine aminotransferase (ALT), aspartateaminotrnsferase (AST) and alka-line phosphatase (ALP) activities after 48 hours and two weeks, while ALT was also significantly increased after one week. Tumor necrosis factoralpha (TNF-α) level was undetected in treated and control groups except the group treated with ZEN for one week. Alphafetoprotein (AFP) level was increased significantly only after two weeks. The activity of antioxidants was significantly increased in all groups. ZEN was also found to modify the serum proteins especially gamma-globulin which showed a significant decrease after 48 h and two weeks. Improvement in liver func-tion occurred in the group pretreated with the crude venom, and AFP and antioxidants returned to normal level, while TNF-α level was also undetected. Gamma globulin was significantly increased. The recovery observed in the group which was pretreated with crude venom may related to bradykinin content of this venom which exhibits a hepatoprotective effect. Histological changes in mouse liver coincided with biochemical changes. In conclusion, this study revealed that ZEN induced liver function and structural changes promising an approach for using a crude venom of jellyfish to enhance liver function.
Mycotoxin Detection in Human Samples from Patients Exposed to Environmental Molds  [PDF]
Dennis G. Hooper,Vincent E. Bolton,Frederick T. Guilford,David C. Straus
International Journal of Molecular Sciences , 2009, DOI: 10.3390/ijms10041465
Abstract: The goal of this study was to determine if selected mycotoxins (trichothecenes, aflatoxins, and ochratoxins) could be extracted and identified in human tissue and body fluids from patients exposed to toxin producing molds in their environment. Human urine and methanol extracted tissues and sputum were examined. Trichothecenes were tested using competitive ELISA techniques. Aflatoxins B1, B2, G1, and G2, and ochratoxin A were tested by using immunoaffinity columns and fluorometry. Test sensitivity and specificity were determined. Levels of detection for the various mycotoxins varied from 0.2 ppb for trichothecenes, 1.0 ppb for aflatoxins, and 2.0 ppb for ochratoxins. Trichothecene levels varied in urine, sputum, and tissue biopsies (lung, liver, brain) from undetectable (<0.2 ppb) to levels up to 18 ppb. Aflatoxin levels from the same types of tissues varied from 1.0 to 5.0 ppb. Ochratoxins isolated in the same type of tissues varied from 2.0 ppb to > 10.0 ppb. Negative control patients had no detectable mycotoxins in their tissues or fluids. These data show that mycotoxins can be detected in body fluids and human tissue from patients exposed to mycotoxin producing molds in the environment, and demonstrate which human tissues or fluids are the most likely to yield positive results.

Wang Jinglin Zhang Zhidong Li Zhongrun Gu Liangyuan Yin Shixing,

菌物学报 , 1994,
Abstract: Six stabilized hybridoma cell lines secreting anti-zearalenone (ZEN) monoclonal antibodies (IgG) were produced by fusing the mouse cells (SP2/0) with splenocytes isolated from Balb/c mice. The Balb/c mice were previously immunized with ZEN-bovine serum albumin (BSA) conjugation which was prepared by ZEN-oxime with BSA. An indirect competitive ELISA was used to detect ZEN by anti-ZEN McAb. Cross reactivity of these antibodies with zearalenol was 1.3-9.0%. The sensitivity of CI-ELISA for ZEN was 0.3-0.8ng/ml. Average recovery rates of ZEN spiked (25-500ng/g) corn, feed and wheat were 105%, 103% and 90%, respectively. Mean coefficients of variation were 3.8%, 15.2% and 12.7% for interwell and 5.8%, 6.8% and 2.8% for interassay, respectively.
Effects of zearalenone in prepubertal gilts
Teixeira, Letícia C.;Montiani-Ferreira, Fabiano;Locatelli-Dittrich, Rosangela;Santin, Elizabeth;Alberton, Geraldo C.;
Pesquisa Veterinária Brasileira , 2011, DOI: 10.1590/S0100-736X2011000800004
Abstract: prepubertal gilts were fed with a diet containing zearalenone (zea) in a concentration of 0.75 mg/kg for 21 days. the effects of this mycotoxin on morphologic aspects of the reproductive tract as well as on complete blood count (cbc), serum biochemistry analysis (sba) and humoral immune response against sheep red blood cells (srbc) were evaluated. there was a significant increase (p<0.05) on the reproductive tract weight, vulvar area, height of the epithelial cells of endometrial glands and uterine mucosa. these results showed the ability of this nonsteroidal mycotoxin in mimicking actions of 17β estradiol at the concentration of 0.75mg/kg. no changes in weight gain, cbc, sba parameters and humoral response against srbc were observed.
Hepatic hyperplasia and damages induces by zearalenone Fusarium mycotoxins in BALB/c mice
Chatopadhyay, Pronobesh;Pandey, Anurag;Chaurasia, Asshwani K.;Upadhyay, Addesh;Karmakar, Sanjev;Singh, Lokendra;
Arquivos de Gastroenterologia , 2012, DOI: 10.1590/S0004-28032012000100013
Abstract: context: zearalenone is a mycoestrogen and considered a mycotoxin. objective: to establish whether zearalenone produced hepatotoxicity via oral administration. methods: zearalenone was orally administered at a dose of 50 mg, 100 mg and 200 mg zen/body weight/daily, respectively, for 14 days to three groups of balb/c mice. diagnostic modalities used to evaluate hepatic damage and impaired hepatic function pre- and post zearalenone administration included hepatic marker enzyme activity, pentobarbital sleeping time, cytochrome p-450 activities and histopathologic evaluation of liver. results: significant histopathologic changes viz. sinusoidal congestion, cytoplasmic vacuolization, hepatocellular necrosis and neutrophil infiltration were observed after evaluating of liver section from each group after accumulated zearalenone exposure. further, zearalenone exposure increased activities of alanine transaminase, aspartate transaminase and lipid peroxides whereas activities of tissue glutathione and cytochrome p450 were decreased as compared to control mice. zearalenone also increased the sleeping time and decreased sleeping latency after pentobarbital through intraperitoneal route as compared to control mice which indicates that the impairment of hepatic metabolizing enzymes by zearalenone. conclusion: zearalenone is a potential hepatotoxin by oral route.
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