首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 31 毫秒
1.
In the annual national survey (‘Besondere Ernteermittlung’) the quality of German cereals was analysed. The samples for this approach were collected from statistically selected fields. One of the analysed parameters was the content of mycotoxins. Thus, it was possible to get an area-wide survey of the content of mycotoxins under natural growing conditions. In 2003 a total number of 974 samples were analysed consisting of wheat, rye and barley samples. A special mode of choosing the wheat samples ensured also a statistically validated result for the federal states of Germany. In the year 2003 the content of the mycotoxins DON and ZEA in wheat and rye samples was lower than in 2002 referred to the median. On the other hand the frequency of detection of DON was not reduced. In barley, which was analysed for the first time in this kind of study, the level of DON was in between the levels of wheat and rye.  相似文献   

2.
Mycotoxins are toxic secondary metabolites produced by a number of different fungi, and can be present in a wide range of food and feed commodities including cereal grains, oil seeds, dried fruits, apple juice, wine and meat products from animals fed contaminated meal. Many mycotoxins are highly resistant, and survive food processing, and therefore enter the food chain and provide a threat to human health. The optical waveguide lightmode spectroscopy (OWLS) technique has been applied to the detection of Aflatoxin and Ochratoxin in both competitive and in direct immunoassays. After immobilizing the antibody or antigen conjugate for the direct or indirect measurement, respectively, the sensor chip was used in flow-injection analyser (FIA) system. When using non-competitive method, sensor responses were obtained first only at analyte concentrations of 5-10 ng ml(-1). In both cases, the responses were very unstable. For competitive sensor investigation with the sensitized chip first the optimal dilution rate of monoclonal antibodies was determined, for the measurement of Ochratoxin A and Aflatoxin B1 the monoclonal antibody stock solution was diluted to 1 microg ml(-1) and to a 1:400 dilution, respectively. During the competitive measurement standard solutions were mixed with monoclonal antibodies at the appropriate concentration, the mixture was incubated for 1 min and injected into the OWLS system. The sensitive detection range of the competitive detection method was between 0.5 and 10 ng ml(-1) in both cases. After the establishment of the indirect method, barley and wheat flour samples were measured, and the results were in good correlation by those measured by enzyme linked immuno-sorbent assay (ELISA). Regression coefficient between the two methods for Ochratoxin and Aflatoxin was determined as 0.96 and 0.89, respectively.  相似文献   

3.
抗脱氧雪腐镰刀菌烯醇单克隆抗体的制备   总被引:2,自引:0,他引:2  
[目的]小麦赤霉病菌产生的毒素不仅在病害发展过程中具有加重赤霉病的作用,而且污染谷物导致严重的食用安全性问题.由于赤霉病的普遍发生,有必要建立快速、灵敏、有效的毒素检测方法,本试验旨在制备可用于检测被脱氧雪腐镰刀菌烯醇污染的粮谷类特异性单克隆抗体.[方法]本实验首先将脱氧雪腐镰刀菌烯醇(DON)的衍生物3-半琥珀酰-脱氧雪腐镰刀菌烯醇(3-HS-DON-OVA)与卵清蛋白(OVA)采用碳化二亚胺法进行偶联得到人工抗原,以此人工抗原免疫BALB/C小鼠,取该鼠脾细胞与SP2/O鼠骨髓瘤细胞融合,经筛选和克隆,得到了1株能稳定分泌DON抗体的单克隆细胞株(382),并制备单克隆抗体腹水.[结果]经检测382的抗体类型及亚类均为IgG1,其轻链为κ链.腹水通过间接酶联免疫吸附测定效价在1×10-7以上.该单克隆抗体与脱氧雪腐镰刀菌烯醇特异性结合反应的50%抑制质量浓度为29 μg/L,除与3-acetyldeoxynivalenol(3-Ac-DON)的交叉反应率为78.38%,与其他脱氧雪腐镰刀菌烯醇结构类似物无交叉反应.[结论]本实验所制备的单克隆抗体有较高的灵敏度和特异性,具有较好的应用价值.  相似文献   

4.
The aim of the present study was to investigate the occurrence of mycotoxins in commercial dog food, as a basis to estimate the risk of adverse effects. Seventy-six dry dog food samples from 27 producers were purchased from retail shops, supermarkets, and specialized pet food shops in Vienna, Austria. The frequency and levels of deoxynivalenol (DON), zearalenone (ZEA), fumonisins (FUM), ochratoxin A (OTA). and aflatoxins (AF) in dry dog food were determined. Mycotoxin analysis were performed by commercial enzyme-linked immunosorbent assay (ELISA) test kits. Confirmatory analyses were done for DON, ZEA, and FUM by high performance liquid chromatography (HPLC) after extract clean-up with immunoaffinity columns. The correlations between ELISA and HPLC results for DON and ZEA were acceptable and indicated that ELISA could be a simple, low cost, and sensitive screening tool for mycotoxins detection, contributing to quality and safety of pet food. DON was the mycotoxin most frequently found (83% positives; median 308 μg/kg, maximum 1,390 μg/kg). ZEA (47% positives, median 51 μg/kg and maximum 298 μg/kg) and FUM (42% positives, median 122 μg/kg and maximum 568 μg/kg) were also frequently detected in dog food. OTA was less frequently found (5%, median 3.6 μg/kg, maximum 4.7 μg/kg. AF were not detected (<0.5 μg/kg) in any sample. The results show that dry dog food marketed in Vienna are frequently contaminated with mycotoxins (DON > ZEA > FUM > OTA) in low concentrations, but do not contain AF. The high frequency of Fusarium toxins DON, ZEA, and FUM indicates the need for intensive control measures to prevent mycotoxins in dog foods. The mycotoxin levels found in dry dog food are considered as safe in aspects of acute mycotoxicoses. However, repeated and long-time exposure of dogs to low levels of mycotoxins may pose a health risk.  相似文献   

5.
Thefusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZON) were determined in conventionally and organically grown grain harvested 1998 in Thuringia/Germany. A total of 196 wheat samples and 69 rye samples was analysed.In this year with heavy rainfalls during the summer months, high concentrations offusarium mycotoxins were typical in grain grown in Germany, as the DON concentrations found here. DON concentrations in conventionally grown wheat were found to be significantly higher than in organically grown wheat. 69% of the conventionally grown wheat were tested positive, containing a mean concentration of 1540 µg/kg DM. In 54% of the organically grown wheat samples DON was detected with a mean value of 760 µg/kg DM. DON concentration in rye and ZON concentration in wheat showed similar tendencies.The different cultivars of conventionally grown wheat showed large differences in DON contamination.  相似文献   

6.
目的:利用稀土离子作为示踪剂,建立DON/ZEN双标记间接竞争时间分辨荧光免疫分析方法同时检测DON、ZEN。 方法:以DON BSA、ZEN-BSA共包被于固相微孔板,与DON/ZEN标准或样品中的DON、ZEN竞争结合抗DON多抗、抗ZEN单抗,然后分别用稀土离子Eu3+-羊抗兔IgG及Sm3+ 羊抗鼠IgG进行示踪检测,并对建立DON/ZEN-双标记TRFIA进行方法学的考核。结果:DON/ZEN-双标记TRFIA检测灵敏度,DON为0.2 ng/ml、ZEN为0.7 ng/ml,检测范围为:DON 0.2~100 ng/ml,ZEN 0.7~50 ng/ml,批内、批间变异率均小于10%。不同样品添加回收实验表明玉米、小麦样品中DON平均回收率分别为102.8%、98.8%,ZEN平均回收率分别为94.2%、95.7%。DON/ZEN-双标TRFIA检测时,DON与ZEN不相互干扰,该方法特异性好。玉米样品检测结果表明,DON/ZEN双标记TRFIA与单标记DON -TRFIA、ZEN-TRFIA试剂盒结果高度相关,具有较好的一致性,两者检测DON的结果相关系数为0.9760,检测ZEN结果的相关系数为0.9695,结论:DON/ZEN-双标记TRFIA灵敏度高,检测范围宽,重复性、稳定性好,一次检测可同时得到DON、ZEN两个结果,是一种简便、快速、经济、稳定、可进行大批量样品筛查的检测方法。  相似文献   

7.
To assess the potential for mycotoxin contamination of the human food supply following the 1988 U.S. drought, 92 grain food samples were purchased from retail outlets in the summer of 1989 and surveyed for aflatoxin B1, zearalenone, and deoxynivalenol (DON [vomitoxin]) by monoclonal antibody-based competitive enzyme-linked immunosorbent assay (ELISA). Only one sample (buckwheat flour) was found to contain aflatoxin B1 (12 ng/g), whereas zearalenone was found in 26% of the samples at a mean concentration of 19 ng/g. In contrast, the DON ELISA was positive in 50% of the samples at a detection level of 1.0 micrograms/g. Between 63 and 88% of corn cereals, wheat flour/muffin mixes, rice cereals, and corn meal/muffin mixes yielded positive results for DON, whereas 25 to 50% of oat cereals, wheat- and oat-based cookies/crackers, corn chips, popcorn, and mixed-grain cereals were positive for DON. The mean DON content of the positive samples was 4.0 micrograms/g, and the minimum and maximum levels were 1.2 and 19 micrograms/g, respectively. When positive ELISA samples were also analyzed by high-performance liquid chromatography, a strong correlation between the two methods was found. The presence of DON in the two highest samples, corn meal and mixed-grain cereal, which contained 19 and 16 micrograms/g, respectively, was quantitatively confirmed by gas chromatography-mass spectrometry. The results indicated that DON was present in 1989 retail food products at concentrations that exceeded those found in previous market surveys and that have been experimentally associated with impaired animal health.  相似文献   

8.
The accuracy and precision of a commercially available system based on an indirect competitive immunoassay and planar waveguide technology was evaluated for the analysis of deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZEAR), and T-2 toxin in wheat. The system generally performed well at the tested concentrations that were close to the regulatory limits of DON and OTA in wheat. The mean percent recovery of OTA from certified and in-house reference materials ranged from 90 to 111 %, with a relative standard deviation of 8–16 % (at 4.2, 4.9, and 7.0 μg/kg). Mean percent recoveries of DON ranged from 75 to 103 %, with a relative standard deviation of 14–20 % (at 610, 940, and 1300 μg/kg). As analyte concentrations approached the lower limits of the working range of 3 μg/kg OTA and 400 μg/kg DON, the mean percent recoveries and relative standard deviation increased for both DON and OTA. A lack of reference materials precluded a thorough evaluation of the method for the analysis of ZEAR and T-2. The particular strength of the technology was that multiple mycotoxins were analyzed simultaneously.  相似文献   

9.
To assess the potential for mycotoxin contamination of the human food supply following the 1988 U.S. drought, 92 grain food samples were purchased from retail outlets in the summer of 1989 and surveyed for aflatoxin B1, zearalenone, and deoxynivalenol (DON [vomitoxin]) by monoclonal antibody-based competitive enzyme-linked immunosorbent assay (ELISA). Only one sample (buckwheat flour) was found to contain aflatoxin B1 (12 ng/g), whereas zearalenone was found in 26% of the samples at a mean concentration of 19 ng/g. In contrast, the DON ELISA was positive in 50% of the samples at a detection level of 1.0 micrograms/g. Between 63 and 88% of corn cereals, wheat flour/muffin mixes, rice cereals, and corn meal/muffin mixes yielded positive results for DON, whereas 25 to 50% of oat cereals, wheat- and oat-based cookies/crackers, corn chips, popcorn, and mixed-grain cereals were positive for DON. The mean DON content of the positive samples was 4.0 micrograms/g, and the minimum and maximum levels were 1.2 and 19 micrograms/g, respectively. When positive ELISA samples were also analyzed by high-performance liquid chromatography, a strong correlation between the two methods was found. The presence of DON in the two highest samples, corn meal and mixed-grain cereal, which contained 19 and 16 micrograms/g, respectively, was quantitatively confirmed by gas chromatography-mass spectrometry. The results indicated that DON was present in 1989 retail food products at concentrations that exceeded those found in previous market surveys and that have been experimentally associated with impaired animal health.  相似文献   

10.
制备Asia I口蹄疫病毒vp2单克隆抗体(mAb)并建立了单抗竞争ELISA方法。用纯化的Asia I型口蹄疫病毒vp2重组蛋白免疫BALB/c小鼠, 将免疫小鼠的脾细胞与骨髓瘤SP2/0细胞融合, 采用间接ELISA和有限稀释法筛选杂交瘤细胞。分别用ELISA、Western blotting检测mAb腹水的效价及其特异性。筛选到杂交瘤细胞2株, 腹水效价均在100×29以上; 以纯化后的Asia I型口蹄疫病毒vp2重组蛋白作为抗原, 利用Asia I型口蹄疫病毒vp2单抗酶标物建立了竞争ELISA方法用来检测Asia I型口蹄疫抗体。临床应用表明, 该方法与UBI公司的口蹄疫全病毒抗体检测试剂盒总符合率达89.0%, 和荷兰赛迪公司的口蹄疫病毒LPB-ELISA抗体检测试剂盒总符合率达86.5%。  相似文献   

11.
Fungal contamination of plant products is an important risk factor for health, because of the high mycotoxin potential deriving from these contaminations with multiple effects: hepatic toxicity, teratogenic, mutagenic and carcinogenic. The contamination of cereals with mycotoxins has been a serious problem in Balkan communities. Several studies implicated mycotoxins, in endemic kidney disease geographically limited to Balkan region (Balkan endemic nephropathy). The trichothecenes are of particular concern because they are ubiquitous found in wheat, corn and barley throughout the world. Fumonisins have been isolated from certain Fusarium species of which FB1, FB2 and FB3 are the major ones produced in naturally contaminated foods.These mycotoxins are produced on cereal grains infected by Fusarium while being grown in-the-field. The aim of this study is to evaluate the presence of the Fusarium species in cereals from West side of Romania and to determinate the concentrations of deoxynivalenol (DON) and fumonisine (F1+F2). Identification of Fusarium species was done using the total number of fungal species determination method. The level of mycotoxins was determined with the immune-enzymatic method ELISA. 27 cereal samples from rural households in three counties in West Romania were analysed.  相似文献   

12.
Biolayer interferometry allows for the real time monitoring of the interactions between molecules without the need for reagents with enzymatic, fluorescent, or radioactive labels. The technology is based upon the changes in interference pattern of light reflected from the surface of an optical fiber when materials bind to the tip of the fiber. The technique represents an alternative to technologies such as surface plasmon resonance, with an advantage in that the flow of extracts through small capillaries is not required. In this report, a deoxynivalenol-bovine serum albumin (DON-BSA) conjugate was non-covalently immobilized to the surface of aminopropylsilane sensors and the change in interference pattern resulting from the binding of DON-specific antibodies was measured. The basis for the assay was the competition between DON and the immobilized DON-BSA for binding to limited amounts of antibody. The technique was used to measure DON in extracts of spiked whole wheat flour, with a limit of detection of 0.10 mg DON/kg. Matrix interferences were an issue, and adequate quantification required using matrix-matched standards. When samples were tested with sensors that had not been conditioned to remove loosely attached DON-BSA, the recoveries at five spiking levels over the range from 0.2 to 5 mg/kg averaged 108.8% [relative standard deviation (RSD) 16.0%]. Using sensors that had been conditioned lowered the average recovery (101.4%) and improved the RSD (13.2%). This suggests that conditioning the sensors helped reduce a bias in the assay towards overestimation. These results, and the ease with which assays can be conducted, suggest further exploration of this technology for detection of mycotoxins is warranted.  相似文献   

13.
Deoxynivalenol (DON) is one of most widespread mycotoxins in cereal commodities, and animal feed is prevalently contaminated at high concentrations. This poses a problem in animal nutrition as especially pigs are very sensitive to DON. An effective process for the reduction of the DON concentration is the treatment of contaminated feed with sodium bisulfite (SBS) whereby DON is transformed into DON-sulfonate (DONS). Although the success of this treatment has been confirmed in several feeding studies, it is unexplained if the decrease of DON is accompanied with a coincident increase of DONS. For this reason, we developed a method for the analysis of DONS using hydrophilic interaction chromatography coupled to tandem mass spectrometry. In order to investigate the correlation between DON and DONS concentrations during SBS-treatment, DON-contaminated wheat was treated with SBS and stored for up to 36 days. At defined timepoints of this treatment, samples were analyzed for DON and DONS using stable isotope labeled standards. The preparation, purification, and structure elucidation of DONS, and the HILIC-HPLC-MS/MS method for the analysis of DONS as well as the results of two storage experiments are presented in this paper.  相似文献   

14.
Experimental field trials were carried out to study the influence of N-fertilization on deoxynivalenol (DON) contamination of winter wheat. Within four years of investigation, no definite effect of mineral N-input at dosages varying between 0 and 240 kg N/ha could be observed on DON concentration in wheat grain. The main factors affecting DON contamination of wheat were theFusarium infection pressure, the weather conditions and the susceptibility of the wheat varieties againstFusarium head blight. DON was analyzed with enzyme-linked immunosorbent assay (ELISA) and, for comparison, some of the positive samples were additionally analyzed with high performance liquid chromatography (HPLC). There was a good correlation between the ELISA and the HPLC results for DON concentration in wheat.  相似文献   

15.
Fusarium mycotoxins such as deoxynivalenol (DON) can occur in cereals conjugated to glucose and probably also to other sugars. These conjugates, which are often referred to as ??masked mycotoxins??, will not be detected with routine analytical techniques. Furthermore, it is suspected that the parent toxin may again be released after hydrolysis in the digestive tracts of animals and humans. Today, our knowledge of the occurrence of these compounds in cereal grains is limited. In this paper, a LC-MS/MS method for the simultaneous determination of DON, deoxynivalenol-3-??-d-glucoside (DON-3-glucoside), 3 acetyl-DON, nivalenol, fusarenon-X, diacetoxyscirpenol, HT-2 toxin, and T-2 toxin in naturally (n?=?48) and artificially (n?=?30) contaminated cereal grains (wheat, barley, oat, rye triticale) is reported. The method has also been applied to whole fresh maize plant intended for production of maize silage (n?=?10). The samples were collected from the harvest years 2006?C2010, The results show that DON-3-glucoside and DON co-occurred in cereal grains and, especially in several of the highly contaminated samples, the concentration of the glucoside can be relatively high, corresponding to over 37?% of the DON concentration. The DON-3-glucoside levels in both the naturally and in the artificially grain inoculated with Fusarium were second only to DON, and were generally higher than those of the other tested trichothecenes, which were found at low concentrations in most samples, in many cases even below the detection limit of the method. This argues for the importance of taking DON-3-glucoside into account in the ongoing discussion within the European Community concerning exposure re-evaluations for setting changed values for the tolerable intake for DON. Our results indicate that, in the naturally contaminated grains and in the Fusarium infested cereal grains (winter and spring wheat, oat, triticale), the concentration level of DON-3-glucoside is positively correlated to the DON content. When the DON concentration is high, then the content of DON-3-glucoside will most probably also be high and vice versa.  相似文献   

16.
The wheat supply is known to have levels of deoxynivalenol (DON) from fungal infections. The fungal infections can be suppressed with benomyl (BEN). Under certain exposure laboratory conditions DON and BEN are developmental toxins. Since use of BEN may result in concurrent dietary exposure to both compounds, evaluation of the relative developmental risks is needed. The dietary NOAELs for pup decreased body weight for DON and BEN are 0.375 mg/kg/day in mice and 5 mg/kg/day in rat, respectively. Human exposure to DON can be estimated from surveys of wheat grain and finished products which gave a mean DON concentration of 0.86 ppm. The tolerance level for BEN in grain is 0.2 ppm. Presence of DON and BEN in wheat at these concentrations would not impact wheat consumption. A modified margin of exposure (modMOE) was developed using “a”; kg (food) /kg (body weight) as the constant for wheat consumption. The modMOE for DON is 28/a and that for BEN is 1667/a. Comparative relative risks can be expressed as the ratio of BEN risks to DON risks (60). Treatment with BEN at the most efficacious time decreased DON levels to 34% of controls, and decreased the comparative risk ratio from 60 to 18. The potential for fungicides to decrease mycotoxin health risks is important in the safety assessment for foodstuffs. This approach improves on current practices for food safety assessment by including the expected reductions in risks from naturally occurring mycotoxins in the regulatory analysis.  相似文献   

17.
禾谷镰刀菌是小麦赤霉病的主要致病菌,其真菌次生代谢产生的单端孢霉烯类B型毒素,如雪腐镰刀菌烯醇(nivalenol,NIV)、脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)和其它乙酰化衍生物等污染小麦籽粒后对人畜健康构成威胁。综述了近年来国内外对小麦赤霉病镰孢菌单端孢霉烯类B型毒素生物合成的主要途径及分子调控研究进展,对毒素合成过程中的重要调控基因如TRI5、TRI7和TRI13在农业中的应用进行了阐述。  相似文献   

18.
The purpose of this study was to develop an LC/MS assay to accurately detect three mycotoxins produced by Fusarium graminearum in various matrices. Using different LC conditions, deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-ADON), and zearalenone (ZEN) were detected in four different matrices (fungal liquid cultures, maize grain, insect larvae and pig serum). The sensitivity of MS detection allowed us to detect concentrations as low as 8 ppb of DON and 12 ppb of ZEN. A very small quantity of matrix was therefore necessary for successful analysis of these toxins and a variety of experimental situations were successfully investigated using this technique. Production of 15-ADON and butenolide was monitored in a liquid culture of F. graminearum under controlled conditions. Using simple extraction procedures, the differential accumulation of DON and 15-ADON was followed in inoculated maize genotypes varying in susceptibility to F. graminearum. Toxicokinetic studies were carried out with maize insect pests reared continually on artificial diets containing ZEN and suggested that larvae may possess the ability to degrade ZEN. Finally, persistence of DON was assessed in pigs fed diet supplemented with DON, results indicated that DON accumulates quickly in pig blood and then levels decline progressively for 12 hours thereafter. The LC/MS study reported here is very useful and flexible for the detection of these mycotoxins in different media and at very low concentrations.  相似文献   

19.
Antibodies specific for deoxynivalenol (DON) were prepared by immunizing rabbits with a deoxynivalenol-hemiglutaryl-HSA conjugate. The antibody showed cross-reactivity with DON, 15a-acetyl-DON (15-Ac DON), and 3a-acotyl-DON (3-Ac DON) of 100%, 216%, and 260%, respectively. No cross-reactivity was observed against 20 further trichothecenes of the A-, B-, and C-type. An indirect competitive ELISA procedure was set up for the detection of DON in liquid matrices occurring in the brewing process, Including beer. ELISA was sensitive down to 3.8 ng/mL. Detection limit for DON in tenfold diluted beer was 50 ppb. Recovery of the toxin in spiked beer was 45%, 77%, and 83% at toxin levels of 50; 500; and 1,000 ppb, respectively. ELISA results corresponded quite well with HPLC data. 196 commercial beers from several German breweries, including 36 gushing-positive samples, were assayed for DON. Screening revealed toxin levels from not detectable to 569 ppb. 69% of the gushing beers had marked levels of DON. 37% of the gushing-negative beer samples contained DON in detectable amounts with a maximum content of 172 ppb. Wheat beers had significantly higher DON-contents than beers derived from barley. Concentration of the toxin was significantly higher in gushing beers than in nongushing beers from both cereal sources.  相似文献   

20.
An investigation for the occurrence of nivalenol (NIV), deoxynivalenol (DON) and zearalenone (ZEN) in cereals (ten wheat, one rye and one corn) harvested in Canada have been carried out using a procedure, which is rapid and sensitive for Fusarium mycotoxins. NIV, DON and ZEN were detected in 4, 9 and 9 out of ten wheat samples, and their average levels in the positives were 23 ng/g, 1257 ng/g and 9 ng/g, respectively. One rye and one corn were also contaminated with a minor amount of NIV. This is the first evidence for the natural occurrence of NIV in cereals grown in Canada, though its level was far less than DON.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号