Dietary exposure assessment of deoxynivalenol in wheat flour and its products sold in Shanghai / 上海预防医学

ObjectiveTo investigate the pollution level of deoxynivalenol （DON） in wheat flour and its products sold in Shanghai， and to assess the health risks of DON exposure for residents in Shanghai who ingested DON from wheat flour and its products. MethodsRisk monitoring data of DON in wheat flour and its products sold in Shanghai from 2017 to 2021 were combined with the consumption data of wheat flour and its products by Shanghai residents. A probabilistic assessment method was used to assess dietary exposure of DON in wheat flour and its products. ResultsThe overall detection rate of DON in wheat flour and its products was 77.3% （1 041/1 347）， with a mean concentration of 226.3 μg·kg-1， P50 of 130.0 μg·kg-1 and a maximum value of 3 080.0 μg·kg-1. The mean daily exposure and 95th percentile daily exposure （by body weight） of DON from wheat flour and its products in Shanghai residents were 0.279 μg·kg-1 and 1.146 μg·kg， accounting for 27.9% and 114.6% of the daily tolerable intake of DON TDI， 1 μg·kg， respectively. The probability assessment results indicated that 6.1% of the whole population in Shanghai had DON exposure exceeding the TDI value. Among them， 12.8% of the population aged 6 years old and below， 16.4% of the population aged between 7 and 17 years old， 3.9% of the population aged between 18 and 59 years old and 3.2% of the population aged 60 years old and above exceeded the TDI value for daily DON exposure through wheat flour and its products. ConclusionCertain populations in Shanghai may face certain health risks from daily DON intake wheat flour and its products. Special attention should be paid to the health risk of daily DON exposure through wheat flour and its products for individuals age below 18 years old .

T-2 toxin/DON induced differentially expressed genes in human chondrocytes and its relationship with Kashin-Beck disease / 中华地方病学杂志

Objective:To investigate the relationship between T-2 toxin, deoxynivalenol (DON) induced differentially expressed genes in human chondrocytes and Kashin-Beck disease (KBD), and to search for potential molecular markers of KBD.Methods:Gene microarray profiling was used to analyze the differentially expressed genes induced by T-2 toxin (0.01 μg/ml) and DON (1.0 μg/ml) in normal human chondrocytes, and the differences and similarities between them and the differentially expressed genes in KBD chondrocytes were compared. KEGG pathway enrichment analysis was performed on differentially expressed genes in each group. And the expression patterns of KBD susceptibility genes in T-2 toxin and DON induced human chondrocytes were further compared and analyzed.Results:Gene microarray profiling analysis showed that there were 882 (349 up-regulated genes, 533 down-regulated genes) and 2 118 differentially expressed genes (1 124 up-regulated genes, 994 down-regulated genes) in human chondrocytes induced by T-2 toxin and DON compared with normal control cells, respectively. Compared with differentially expressed genes in KBD chondrocytes, the genes with the same expression trend included B cell translocation gene 1 (BTG1), G protein signaling regulatory protein 5 (RGS5), fatty acid binding protein 4 (FABP4) and key protein senescence 1 (FBLN1), the same KEGG pathway including p53, extracellular matrix receptor interaction and phosphatidylinositol-3-kinase-protein kinase B (PI3K-Akt) signaling pathway. Both T-2 toxin and DON induced human chondrocytes to up-regulate the expression of KBD susceptibility gene growth differentiation factor 5 (GDF5) and down-regulate the expression of collagen type ⅨA1 (COL9A1).Conclusion:The BTG1, RGS5, FABP4, FBLN1, GDF5 and COL9A1 genes play an important role in the pathogenesis of KBD and can be used as potential molecular markers for the pathogenesis of KBD.

Fungal and Mycotoxin Contamination of Stored Maize in Kogi, Northcentral Nigeria: An Implication for Public Health

The maize value chain in the Kogi State and most parts of the country from where maize is purchased into the State lacks mechanisms that ensure grain quality and safety. Against the above-backdrop, this study was designed to evaluate toxigenic fungi and associated mycotoxins in maize produced within different agro-zones of Kogi State. Harvested and stored maize seeds under different storage conditions were collected from three different zones (Zone B Bassa, Zone C Lokoja, and Zone D Idah) and cultured. Different fungal species were isolated by culturing using the spread plate technique on potato dextrose agar (PDA) and identified microscopically. Mycotoxin production by isolated fungi was subsequently evaluated for Deoxynivalenol (DON) contamination using the High-Performance Liquid Chromatography technique (HPLC). The outcome of the study was statistically analysed using simple frequencies and percentages. Aspergillus spp. and Penicillium spp. were the fungi found to be associated with the stored seeds in Kogi, while Fusarium spp. Mucor spp. and Rhizopus spp. were the field fungi identified. Of the thirteen samples collected, the most common genera were Aspergillus (isolated from 41.67% of the evaluated samples), Fusarium (27%) and in a lesser extent Rhizopus spp. (8.33%). The result also shows DON was detected in 92.3% of the stored maize samples, making it one of the widespread mycotoxin contaminants of maize grain. Implications of this study for human and animal health and economic development were discussed and appropriate recommendations made especially for adoption of proper storage technology among small-scale farmers for improved maize quality and safety.

Exposure status and health risk assessment of deoxynivalenol from cereals in Chinese population in different regions / 中华预防医学杂志

Objective@#To evaluate the dietary exposure to deoxynivalenol (DON) from cereals and health risk in Chinese residents in different regions.@*Methods@#The data of DON concentration in cereals was derived from the national food safety risk surveillance from 2010 to 2017, with 15 422 samples of cereals included. China was roughly divided into north part and south part, along with the Qinling Mountains-Huaihe River line. Sample size of each type of cereals, i.e. wheat flour, maize meal, oats and rice was 4 948, 696, 626, 1 006 in the north, while 5 648, 1 068, 266, 1 164 in the south. The data of cereals consumption was derived from China National Nutrition and Health Survey in 2002 and 68 335 respondents aged 3 and above, with 34 234 from the north and 34 101 from the south, were included. Simple distribution model was applied for calculation and comparison of the dietary exposure to DON from cereals in northern and southern residents based on individual consumption of cereals, body weight and average DON concentration in each type of cereals.@*Results@#Average DON concentration in wheat flour, maize meal, oats, and rice sampled in northern China were 235.4, 121.6, 7.0 and 4.6 μg/kg, respectively, while 239.1, 124.3, 29.0 and 15.5 μg/kg in cereals sampled in southern China. The average DON exposure from cereals in surveyed Chinese inhabitants was 0.78 μg/(kg·d). Among them, the DON exposure of northern residents was higher than that of southern residents (P<0.001), and the average exposures were 1.15 and 0.41 μg/(kg·d), respectively. A total of 49.2% of northern residents exceeded provisional maximum tolerable daily intake for DON exposure from cereals, which was much higher than that of southern residents (8.6%) (P<0.001). Wheat-based food products were the main source of DON exposure, with a contribution rate of 96.5% in the north and 68.3% in the south. Average DON exposure was the highest in the 3-6 years [2.12 μg/(kg·d) for children in north and 0.73 μg/(kg·d) in south].@*Conclusion@#Exposure to DON from cereals in northern residents of China was considerably high, with a certain health risk. Northern children aged 3 to 6 exposed even more DON and needed significant attention.

Local strains Aspergillus oryzae KKB4 and Rhizopus oryzae KP1R1 as a reducing and detoxifying agents for deoxynivalenol

Aims@#Deoxynivalenol is a type B trichothecene produced by Fusarium graminearum that can cause serious health problems in human and livestock. The present study aimed to reduce and detoxify deoxynivalenol using a local strain Aspergillus oryzae KKB4 and Rhizopus oryzae KP1R1. @*Methodology and results@#Corn as solid substrate artificially inoculated with F. graminearum bio 163252 to produce deoxynivalenol. Deoxynivalenol contaminated corn then inoculated with A. oryzae KKB4 and R. oryzae KP1R1. During fermentation, a decrease in deoxynivalenol levels is analyzed including loss of dry matter and glucosamine content. Deoxynivalenol was extracted from the substrate by solid phase extraction and quantified using high-performance liquid chromatography. The reduction of deoxynivalenol by A. oryzae KKB4 and R. oryzae KP1R1 were 65.91% and 56.82%, respectively after ten days of fermentation. Toxicity analysis revealed that residues of deoxynivalenol were not toxic to growth of Saccharomyces cerevisiae cells. @*Conclusion, significance and impact of study@#Local strains A. oryzae KKB4 and R. oryzae KP1R1 were able to reduce and detoxify deoxynivalenol in solid substrates. This study provides supporting data to control mycotoxin that is critical for food and feed safety.

Effects of deoxynivalenol on articular cartilage injury / 中华地方病学杂志

Deoxynivalenol (DON) is one of mycotoxins produced by fungal pathogen of fusarium,which has strong cytotoxicity.The study of articular cartilage injury is focused on the study of Kaschin-Beck disease (KBD)pathogenesis.DON can inhibit the proliferation of chondrocytes,induce apoptosis,affect the cell-related metabolic enzymes and other ways to break the balance of chondrocytes and extracellular matrix,eventually leading to irreversible damage of articular cartilage.This article reviews the role of DON toxin in the cartilage injury.

Determination of mycotoxins in pu-erh tea, black tea, and green tea samples / 中国中药杂志

To establish a robust method for the determination of mycotoxins in tea samples, and to provide means for the quality and safety control of tea products. Samples of 20 tea products acquired from international market were extracted by organic solvents (acetonitrile containing 0.1% formic acid) or hot water, respectively. The extracts were analyzed by UPLC-MS/MS.A good linear regression was achieved in a range of 39.1 to 5 000 ng•L⁻¹ for aflatoxin B1 (AFB1) and aflatoxin G1 (AFG1), 117 to 15 000 ng•L⁻¹ for aflatoxin B2 (AFB2) and aflatoxin G2 (AFG2), 2.44 to 313 ng•L⁻¹ for fumonisin B1 (FB1), fumonisinB2 (FB2) and fumonisin B3 (FB3), and 3 125 to 5 000 ng•L⁻¹ for deoxynivalenol, with recovery rates between 85.7% and 99.6%. The coefficient of the linear equation for all standards was greater than 0.999 0, and the RSD value was less than 10%. Mycotoxins were detected in several tea samples using the two extraction methods but with different outcomes. The levels of mycotoxins detected ranging from 0.15 to 7.31 μg•kg⁻¹ were well below the State or US FDA regulation limits of mycotoxins in food products. Both methods are simple, accurate, and sensitive, and thus, suitable for the quantitative determination of mycotoxins in different food products. The method with the 80 ℃ hot-water extraction is more appropriate to determine the trace amounts of mycotoxins in tea leaves that are likely to be present in brewed tea liquor, while organic solvent method is more suitable for the detection of mycotoxins in ingestible foods.

Evaluation of a mycotoxin adsorbent in swine diets containing barley naturally contaminated with Fusarium mycotoxins / Evaluación de un adsorbente de micotoxinas en dietas para cerdos que contienen cebada contaminada naturalmente con micotoxinas de Fusarium / Avaliação de um adsorvente de micotoxinas em suínos dieta contendo cevada naturalmente contaminado com micotoxinas Fusarium

Summary Background: contamination of barley with Fusarium mycotoxins causes significant economic losses for pork producers, and alleviation with mycotoxin sequestering agents has proven inconsistent. Objective: to evaluate a yeast cell wall product in preventing adverse effects of Fusarium mycotoxins on growth performance, blood characteristic, and nutrient digestibility in gilts fed diets containing barley naturally contaminated with Fusarium mycotoxins. Methods: positive and negative controls of corn-soybean meal diets containing 20% control and contaminated barley with Fusarium mycotoxins, respectively, were prepared. Two additional diets were prepared by adding 0.2 or 0.4% of a yeast cell wall product to the negative control diet. The experimental diets were fed to pigs with 61.7 Kg initial body weight for 2 weeks. Results: pigs fed the negative control diet gained less than those fed the positive control diet (p<0.05) from d 0 to 7 and during the overall period, but nutrient digestibility and blood characteristics were not affected by feeding the contaminated diet. Most measurements were not affected by supplementing the yeast cell wall to the negative control diet. Conclusion: addition of the yeast cell wall product to negative control diets failed to ameliorate the adverse effects of dietary Fusarium mycotoxin on growth performance.

Resumen Antecedentes: la contaminación de la cebada con micotoxinas de Fusarium causa importantes pérdidas económicas para los productores de carne de cerdo, pero adsorbentes de micotoxinas para prevenir los efectos perjudiciales de las micotoxinas no dieron resultados consistentes. Objetivo: evaluar el producto de la pared celular de la levadura en la prevención de los efectos adversos de las micotoxinas de Fusarium sobre el crecimiento, características de la sangre, y la digestibilidad de nutrientes en cerdas jóvenes alimentadas con dietas que contienen cebada naturalmente contaminada con micotoxinas de Fusarium. Métodos: se prepararon las dietas a base de harina de maíz-soja de control positivo y negativo contienen 20% de control y cebada contaminados con micotoxinas de Fusarium, respectivamente. Adicionalmente se prepararon 2 dietas con productos de la pared celular de levadura de 0,2 o 0,4% a la dieta de control negativo. Fueron alimentados con las dietas experimentales cerdos con un peso corporal inicial de 61,7 Kg durante 2 semanas. Resultados: los cerdos alimentados con la dieta control negativo ganaron menos que aquellos alimentados con la dieta control positivo (p<0,05) a partir del d 0 a 7 y durante el período en general, pero la digestibilidad de los nutrientes y características de la sangre no fueron afectados por la alimentación de la dieta contaminada. La mayoría de las mediciones no fueron afectadas por la suplementación de la pared celular de la levadura con la dieta control negativo. Conclusión: la adición del producto de pared celular de levadura a la dieta control negativo no logró aminorar los efectos adversos de las micotoxinas de Fusarium sobre el crecimiento.

Resumo Antecedentes: a contaminação da cevada com a micotoxinas Fusarium causa perdas econômicas significativas para os produtores de suínos, porèm os adsorventes de micotoxinas que evitam efeitos prejudiciais de micotoxinas não deram resultados consistentes. Objetivo: avaliar o produto da parede celular das leveduras para prevenir efeitos adversos da micotoxinas Fusarium no desempenho do crescimento, inchaço da vulva, característica do sangue, digestibilidade dos nutrientes em porcas jòvens que são alimentadas com dietas contendo a cevada naturalmente contaminada com micotoxinas do Fusarium. Métodos: foram preparadas dietas de controle positivo e negativo à base de milho e soja, contendo 20% do controle e cevada contaminada com micotoxinas Fusarium, respectivamente. Duas dietas adicionais foram preparadas complementando com 0,2 ou 0,4% o produto da parede celular das leveduras à dieta do controle negativo. As dietas experimentais foram fornecidas aos suínos com peso corporal inicial de 61,7 Kg por 2 semanas. Resultados: os suínos alimentados com a dieta do controle negativo ganharam menos peso do que aqueles alimentados com a dieta do controle positivo (p<0,05) a partir do dia 0 até 7 durante todo o período, a digestibilidade dos nutrientes e as características do sangue não foram afetadas pela alimentação da dieta contaminada. A maioria das medidas não foram afetadas pela complementação da parede celular das leveduras à dieta do controle negativo. Conclusão: a complementação do produto da parede celular das leveduras nas dietas de controle negativo não conseguiu melhorar o efeito adverso das micotoxinas Fusariume no desempenho de crescimento.

Preparation and Application of Immunoaffinity Column for Multi-mycotoxins / 分析化学

The three-in-one immunoaffinity column ( IAC ) for the determination of aflatoxin B1 ( AFB1 )-zearalenone (ZEN)-deoxynivalenol (DON) was prepared with rProtein A-sepharose 4B as the column matrix. The comprehensive performance ( such as nonspecific adsorption, column blank, column capacity, column efficiency and sample standard addition recovery rate) was evaluated and investigated. The results showed that, the column capacities of AFB1 , ZEN, DON were 295 ng per 0. 25 mL gel, 905 ng per 0. 25 mL gel, 2342 ng per 0. 25 mL gel, respectively, and the column blank was 0. The average recoveries of AFB1 , ZEN and DON were 97. 4%, 98. 0% and 98. 4%, respectively. By optimizing conditions, the samples were extracted using the mixture of methanol and water (80:20, V/V), and diluted with phosphate buffered saline (contain 0. 1% Tween-20, PBST). The detection results of FAPAS (Food Analysis Performance Assessment Scheme) by different batch three-in-one columns were close to the target value. The prepared three-in-one immunoaffinity column which could take the place of conventional single immunoaffinity column was able to meet the requirement for treatment of food and feed samples, and lay a foundation for one step enrichment, purification and detection of multi-mycotoxins.

Safety issues and new rapid detection methods in traditional Chinese medicinal materials

The safety of traditional Chinese medicine (TCM) is a major strategic issue that involves human health. With the continuous improvement in disease prevention and treatment, the export of TCM and its related products has increased dramatically in China. However, the frequent safety issues of Chinese medicine have become the 'bottleneck' impeding the modernization of TCM. It was proved that mycotoxins seriously affect TCM safety; the pesticide residues of TCM are a key problem in TCM international trade; adulterants have also been detected, which is related to market circulation. These three factors have greatly affected TCM safety. In this study, fast, highly effective, economically-feasible and accurate detection methods concerning TCM safety issues were reviewed, especially on the authenticity, mycotoxins and pesticide residues of medicinal materials.

Determination of Deoxynivalenol in Four Traditional Chinese Herbal Medicines by LC-MS-MS / 中国药师

Objective:To develop a method for determining deoxynivalenol in four kinds of traditional Chinese herbal medicines. Methods:After being extracted by water, purified by an immunoaffinity column, deoxynivalenol was analyzed by LC-MS-MS. Results:The calibration curve was linear within the range of 2-50 ng·ml-1 for deoxynivalenol. The recovery was 68. 7%-88. 3%. Conclusion:The method is simple, sensitive and accurate in the determination of deoxynivalenol in Semen Ziziphi Spinosae, Semen Sojae Praepara-tum, Semen Coicis and Fructus Psoraleae.

Incidence and multiplex PCR based detection of trichothecene chemotypes of Fusarium culmorum isolates collected from freshly harvested Maize kernels in Southern India

Hundred Fusarium culmorum strains, isolated from freshly harvested maize grain samples from Southern parts of India, were incubated in czapek-dox medium and analyzed for trichothecene (DON/NIV) production. The mPCR assay was standardized targeting trichothecene metabolic pathway genes viz., Tri6, Tri7, Tri13 for detection of trichothecene (DON/NIV) chemotypes and rDNA gene for specific detection of F. culmorum species. Primers for targeted genes were designed and used to predict whether these isolates could produce deoxynivalenol/nivalenol, 94 isolates were able to produce DON/NIV by mPCR assay. Chemical analysis of DON/NIV was carried out for mPCR positive isolates by high performance-thin layer chromatography (HPTLC). To check the practical usefulness of developed mPCR assay, 150 field samples of maize were evaluated and results were compared with conventional HPTLC method. Out of 150 samples, 34% samples stayed as a positive for NIV contamination whereas 44% were found to have deoxynivalenol contamination. Moreover, mPCR results are equivocally matched with the HPTLC chemical analysis for field samples. Chemotyping of F. culmorum isolates were reported for the first time from India, and highlights the important potential of F. culmorum to contaminate maize with DON/NIV.

Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay

Monoclonal antibody (mAb, NVRQS-DON) against deoxynivalenol (DON) was prepared. DON-Ag coated enzyme linked immunosorbent assay (ELISA) and DON-Ab coated ELISA were prepared by coating the DON-BSA and DON mAb. Quantitative DON calculation ranged from 50 to 4,000 ng/mL for DON-Ab coated ELISA and from 25 to 500 ng/mL for DON-Ag coated ELISA. 50% of inhibitory concentration values of DON, HT-2, 15-acetyl-DON, and nivalenol were 23.44, 22,545, 5,518 and 5,976 ng/mL based on the DON-Ab coated ELISA. Cross-reactivity levels of the mAb to HT-2, 15-acetyl-DON, and nivalenol were 0.1, 0.42, and 0.40%. The intra- and interassay precision coefficient variation (CV) were both <10%. In the mAb-coated ELISA, mean DON recovery rates in animal feed (0 to 1,000 microg/kg) ranged from 68.34 to 95.49% (CV; 4.10 to 13.38%). DON in a buffer solution (250, 500 and 1,000 ng/mL) was isolated using 300 microg of NVRQS-DON and 3 mg of magnetic nanoparticles (MNPs). The mean recovery rates of DON using this mAb-MNP system were 75.2, 96.9, and 88.1% in a buffer solution spiked with DON (250, 500, and 1,000 ng/mL). Conclusively we developed competitive ELISAs for detecting DON in animal feed and created a new tool for DON extraction using mAb-coupled MNPs.

Technetium-99m-labeled deoxynivalenol from Fusarium mycotoxin alters organ toxicity in BALB/c mice by oral and intravenous route

The toxicity of deoxynivalenol, both intravenously and orally, was investigated in male and female BALB/c mice. Technetium-99m (99m Tc)-labeled deoxynivalenol was administered to mice by tail vein injection and orally dosed. Distribution of labeled deoxynivalenol at 26 hours was monitored by gamma-scintigraphy. In the evaluated organs, the accumulation of radioactive deoxynivalenol was correlated with the amount of radioactivity. In addition, the toxicity of deoxynivalenol was measured by biochemical assays followed by histopathological findings. Kidney and hepatic marker enzymes were significantly increased in intravenously administered deoxynivalenol as compared to orally treated mice. Intravenously treated mice showed severe damage in liver and kidney when compared to those orally exposed. Biodistribution of 99mTc-labeled deoxynivalenol differed between oral and intravenous treatment. In intravenously exposed mice, deoxynivalenol was distributed primarily in the liver and kidney whereas in oral exposure, it was found in the stomach and intestines after 26 hours. Deoxynivalenol toxicity, associated with its biodistribution and organ toxicity, was greatest where it had accumulated. The results show that the toxicity of deoxynivalenol is associated with organ accumulation.(AU)

Induction of apoptotic lesions in liver and lymphoid tissues and modulation of cytokine mRNA expression by acute exposure to deoxynivalenol in piglets

Six 1-month-old piglets were intravenously injected with deoxynivalenol (DON) at the concentration of 1 mg/kg body weight, with three pigs each necropsied at 6 and 24 h post-injection (PI) for investigation of hepatotoxicity and immunotoxicity with special attention to apoptotic changes and cytokine mRNA expression. Histopathological examination of the DON-injected pigs revealed systemic apoptosis of lymphocytes in lymphoid tissues and hepatocytes. Apoptosis of lymphocytes and hepatocytes was confirmed by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method and immunohistochemical staining against single-stranded DNA and cleaved caspase-3. The number of TUNEL-positive cells in the thymus and Peyer's patches of the ileum was increased at 24 h PI compared to 6 h PI, but the peak was at 6 h PI in the liver. The mRNA expression of interleukin (IL)-1beta, IL-6, IL-18, and tumor necrosis factor (TNF)-alpha in the spleen, thymus and mesenteric lymph nodes were determined by semi-quantitative RT-PCR, and elevated expression of IL-1beta mRNA at 6 h PI and a decrease of IL-18 mRNA at 24 h PI were observed in the spleen. IL-1beta and IL-6 mRNA expressions increased significantly at 6 h PI in the thymus, but TNF-alpha decreased at 6 h PI in the mesenteric lymph nodes. These results show the apoptosis of hepatocytes suggesting the hepatotoxic potential of DON, in addition to an immunotoxic effect on the modulation of proinflammatory cytokine genes in lymphoid organs with extensive apoptosis of lymphocytes induced by acute exposure to DON in pigs.

Validity of method for determining deoxynivalenol through high performance liquid chromatography with detection in the ultraviolet in wheat samples / Validación de un método para la determinación de deoxinivalenol por cromatografía líquida de alta resolución con deteccíon ultravioleta en muestras de trigo

With the purpose of evaluating the wheat grain and wheat flour contamination by Deoxynivalenol (DON) in the municipality of Chapeco-SC, and standardize an useful method to detect this mycotoxin by High Performance Liquid Chromatography (HPLC), six samples of wheat grains from different storage location, and one sample of wheat grain from the flour milling industry were obtained during in the month of august 2008. Samples belong to a corporation from Chapeco-SC that stores and processes wheat grains, flour and wheat middlings, among other products. The extraction has been carried out with methanol: water (100: 100 v / v), filtered paper filter and applied in immunoaffinity column specific DON. After the wash with water column, the toxin was eluted with methanol. The detection and quantification Deoxynivalenol in samples was carrid out through the method of HPLC in the UV-visible with detection 244 nm. The 6 analyzed samples of wheat grain showed DON levels within 7.0 and 10.1 ppb, while the wheat flour contained 90.2 ppb. DON contents in wheat grains and wheat flour are lower than the limits claimed by the studied corporated importers and the international legislation.

Con el objetivo de evaluar la contaminación por Deoxinivalenol (DON) en granos y harina del trigo en la municipalidad de Chapeco-SC y estandarizar un método de deteccíon para este micotoxina por cromatografía líquida de alta resolución (CLAR), se procesaron durante el mes de agosto de 2008, seis muestras de granos del trigo en diferentes situaciones de almacenamiento y una muestra de harina de trigo de un molino . Las muestras pertenecen a una cooperativa de Chapeco-SC que procesa y almacena granos y harinas de trigo entre otros productos. La extracción de la micotoxina se obtuvo con metanol: agua (100: 100 v / v), filtrado en papel filtro y aplicado a una columna de inmunoafinidad específica (DON). Después del lavado de la columna con agua, la toxina fue elucidada con metanol. La detección y cuantificación de Deoxinivalenol en las muestras se determinó por el método CLAR en el UV- visible con una longitud de onda de 244 nm. Las 6 muestras analizadas del grano, mostraron que el DON nivela entre 7,0 y 10, 1 ppb, mientras la harina del trigo alcanzó las 90,2 ppb. Los niveles de DON en los granos y harina de trigo tienen límites menores que los exigidos por las cooperativas importadoras estudiadas y la legislación internacional.

Investigation on DNA damage of mouse mononuclear cells in peripheral blood induced by low-dose T-2 toxin and deoxynivalenol alone or in combination / 中国地方病学杂志

Objective To investigate the characteristics and extent of mononuclear ceils DNA damage in peripheral blood of mice fed with low dose T-2 toxin and Deoxynivalenol(DON) alone or in combination and to explore the long-term toxicity of the toxin at sub-clinical dose. Methods Eighty female Balb/c mice weighing (14.0 ± 1.5)g 3 weeks after birth were divided randomly into control group, T-2 toxin group, DON group and T-2 toxin combined with DON group according to their body weight, 20 in each group. The mice were injected intraperitoneally T-2 toxin(5 μg·kg-1·d-1), DON(20 μg·kg-1·d-1), T-2 toxin(5 μg·kg-1·d-1) combined with:DON (20μg·kg-1·d-1)respectively,control group were treated by isotonic NaCl. In 16 weeks and 21 weeks of exposure, the tail blood of the mice was collected. The comet rate, tail DNA content,tail length and tail extent moment of mouse mononuclear ceils in peripheral blood was observed using single cell gel electrophoresis(SCGE). Results ① In T-2 toxin group,tail DNA content,tail length and tail extent moment were (27.71 ± 15.85)%, (13.67 ± 5.56)μm, 4.26 ± 3.83 at 16 weeks and (28.38 ± 15.57)%, (13.83 ± 5.47)μm, 4.37 ± 3.82 at 21 weeks, all levels of the indexes increased. In the control group, the corresponding values were (11.87 ± 4.61)%, (10.59±6.70)μm, 1.34±0.98 at 16 weeks and (11.31 ± 3.94)%, (10.83 ± 7.05)μm, 1.29±1.01 at 21 weeks, the differences in the two groups were significant (all P < 0.05) ;②In DON group, the comet rate of cells, tail DNA content and tail extent moment of comet ceils were 5.62%, (28.13 ±13.31)%, 3.39 ± 2.35 at 16 weeks and 7.71%, (29.17 ± 15.12)%, 5.70 ± 4.17 at 21 weeks. In the control group, the tailing rate was 4.34% at 16 weeks and 4.38% at 21 weeks, the differences in the two groups were significant (all P < 0.05);③In the group of T-2 toxin combined with DON,the comet rate, tail DNA content, tail length and tail extent moment was 6.21%, (30.14 ± 15.48)%, (16.93± 6.58)μm, 5.54 ± 4.22 at 16 weeks and 8.17%, (30.85 ± 15.76)%, (17,21±6.45)μm, 5.70 ± 4.17 at 21 weeks. Moreover, the levels were significantly higher than that in the control group(all P < 0.05). The tail DNA content and length of comet cell tail significantly increased in the combine group compared with T-2 group or DON group(P < 0.05). Conclusions Low dose T-2 toxin or DON can definitely result in DNA damage of mononuclear cells in peripheral blood of mice. The damage induced by T-2 toxin combined with DON is severer than that caused by T-2 toxin or DON alone.

Experimental observation of articular impairment of New Zealand rabbits by deoxynivalenol / 中国地方病学杂志

Objective To observe toxic effect of deoxynivalenol(DON)on articular cartilage and synovium of New Zealand rabbits's knee ioints.Methods Fifteen male rabbits were divided randomly into 3 groups:control, high-dosage,and low-dosage group.In high-dosage and low-dosage group,saline solution of DON was injected with a dose of 0.10 and 0.05 ms/kg every 48 h into ear vein of rabbits.Specimen of articular cartilage and synovium were through pathologY methods,and IL-1β,TNF-α,NO levels were assayed in joint liquid,after 20 days. Results Morphological changes were observed, such as synovium inflammative infiltration, chondrocytes deformation and necrosis under light microscope.The levels of IL-1β,TNF-α and NO had statistical significance in comDarison between 3 grouPs(F=19.396,18.195,22.136,P＜0.05).The levels of IL-1β,TNF-α and NO were significantly higher(all P＜0.05),high-dosage[(0.451±0.091),(0.575±0.122)μg/L;(70.27±11.53)μmol/L] and low-dosage group[(0.295±0.107),(0.387±0.131)μg/L;(45.32±12.24)μmol/L]compared with control ((0.1 13±0.049),(0.138±0.087)μg/L;(23.56±9.35)μmoL/L],and high-dosage compared with low-dosage group Conclusions DON results in articular and synovial impairment,which has the symptom similar to osteoarthritis. DON probably causes osteoarthritis.

Effects of deoxynivalenol on apoptosis of human gastric carcinoma cell line SNU in vitro / 肿瘤研究与临床

Objective To explore the effects of deoxynivalenol (DON) on apoptosis of human gastric carcinoma cell line SNU in vitro. Methods SNU cells were treated with DON at different concentrations (50, 100, 1000, 2000 μg/L) for 12 hours, and then cells were harvested for cell apoptosis by flow cytometric (FCM) DNA analysis and the expression of Bax, Bcl-2 and Caspase-3 at protein level with FCM and Western blotting. Results FCM results showed that the apoptosis rates of SNU cells in DON treatment groups were all higher than that in control, especially in DON 1000 μg/L and 2000 μg/L groups (P<0.05). In the concentration range from 50 to 2000 μg/L, a significant concentration-depended response correlation could be found between apoptosis rate and DON concentration (r =0.940, P <0.01). FCM and Western blotting showed Bax and Caspase-3 expression in often SNU cells DON treatment for 12 hours were up-regulated while that of Bcl-2 was down-regulated. Conclusion DON can induce apoptosis of SNU cells in vitro in dose-dependent manner, and possible mechanisms of apoptosis induction effects may be up-regulation of the expression of Bax and down-regulation of that of Bcl-2 and activation of the key enzyme of apoptosis Caspase-3.

Plasma haptoglobin and immunoglobulins as diagnostic indicators of deoxynivalenol intoxication

This study aimed to discover potential biomarkers for dioxynivalenol (DON) intoxication. B6C3F1 male mice were rally exposed to 0.83, 2.5 and 7.5 mg/kg body weight (bw) DON for 8 days and the differential protein expressions in their blood plasma were determined by SELDI - Time-of-Flight/Mass Spectrometry (TOF/MS) and the immunoglobulins (Igs) G, A, M and E in the serum were investigated. 11.7 kDa protein was significantly highly expressed according to DON administration and this protein was purified by employing a methyl ceramic HyperD F column with using optimization buffer for adsorption and desorption. The purified protein was identified as a haptoglobin precursor by peptide mapping with using LC/Q-TOF/MS and MALDI-TOF/MS and this was confirmed by western blotting and ELISA. IgG and IgM in serum were decreased in a dose-dependent manner and IgA was decreased at 7.5 mg/kg bw DON administration, but the IgE level was not changed. To compare the expressions of haptoglobin and the Igs patterns between aflatoxin B1 (AFB1), zearalenone (ZEA) and DON intoxications, rats were orally administered with AFB1 1.0, ZEA 240 and DON 7.5 mg/kg bw for 8 days. Haptoglobin was increased only at DON 7.5 mg/kg bw, while it was slightly decreased at ZEA 240 mg/kg bw and it was not detected at all at AFB1 1.0 mg/kg bw. IgG and IgA were decreased by DON, but IgG, IgA, IgM and IgE were all increased by AFB1. No changes were observed by ZEA administration. These results show that plasma haptoglobin could be a diagnostic biomarker for DON intoxication when this is combined with examining the serum Igs.