ELISA法测定谷物中伏马毒素

为测定谷物中伏马毒素含量，了解粮食中有关真菌的污染状态，评价伏马毒素的ＥＬＩＳＡ测定法。采集市售大麦、玉米及饲料玉米，采用酶联免疫吸附法测定，对阳性样品进行真菌培养分离，并用ＨＰＬＣ进行确证。结果：４０份大米中未检出伏马毒素，７０份玉米检出阳性样品２７份，均数０．６８ｍｇ／ｋｇ。ＨＰＬＣ法的确证结果表明２７份ＥＬＩＳＡ法阳性样品中２３份检出伏马毒素Ｂ１，均数为０．４２ｍｇ／ｋｇ；６份同时检出伏马毒素Ｂ２，均数为０．２ｍｇ／ｋｇ。２７份阳性样品真菌分离培养结果均检出串珠镰刀菌。结论：玉米受串珠镰刀菌污染较重，并可有伏马毒素产生。本次调查大米中未检出伏马毒素，情况较好。其它主粮中伏马毒素污染情况还有待进一步调查     

粮食中伏马菌素污染及产生源相关性研究

目的 对浙江省的723份粮食进行霉菌相分析和伏马菌素及产生源的测定，揭示污染状况和产生源的相关特性。方法 用蔡氏琼脂平板法测定霉菌污染率和侵染率。用ELISA法测定伏马菌素B1。结果 发现样品霉菌污染率达98．6％，侵染率达68．0％，主要菌相是灰绿曲霉、黑曲霉和串珠镰刀菌等，伏马菌素的样品阳性率达72．6％，平均污染量10．5mg/kg。检出的20株串珠镰刀菌全部产毒。结论 经逐步回归统计分析，粮食中的总霉菌污染量、镰刀菌总数各自与伏马菌素污染量呈一元正相关；粮食中伏马菌素污染量与串珠镰刀菌和其它镰刀菌的侵染率共同呈二元正相关，并且其它镰刀菌比串珠镰刀菌对伏马菌素污染量的影响大一倍左右，显示浙江省粮食中伏马菌素的污染具有多源性。     

象山县粮食中杂色曲霉及杂色曲霉素污染调查

杂色曲霉在自然界中广泛存在，并能产生致癌毒素—杂色曲霉素（ＳＴ），国内有报道和胃癌高发有关。本站通过对６０份大米、小麦、玉米样品进行杂色曲霉和杂色曲霉素的测定。结果表明：小麦中杂色曲霉污染严重，污染率达６０％，经产毒鉴定有６０％产毒。大米、玉米中未检出杂色曲霉。三种主粮中杂色曲霉素（ＳＴ）污染较普遍，总污染率为７８．３％，平均污染量２６１．３ｍｇ／ｋｇ，最高达１６２６．０ｍｇ／ｋｇ，其中小麦污染最为严重，污染率依次为小麦１００％，玉米９５％，大米４０％，平均污染量分别为小麦５３４．１ｍｇ／ｋｇ，玉米２７．５ｍｇ／ｋｇ，大米２．４ｍｇ／ｋｇ。     

3种粮食中伏马菌素污染状况调查

串珠镰刀菌分布极广 ,经常污染粮食作物 ,产生多种具有致癌性和急慢性毒性的代谢产物 ,其中伏马菌素 (简称 FB)较为常见且毒性较强 ,成为美国、日本等发达国家继黄曲霉毒素之后的又一个研究热点。我国对 FB的研究尚未全面开展 ,为进一步探讨真菌毒素与人类疾病的关系 ,给制定国家卫生标准提供依据 ,对小麦等 3种粮食中的伏马菌素污染状况进行了调查 ,现将结果报告如下。1　材料与方法1.1　样品来源　选择平原、丘岭、沿海等有代表性的地区 ,分别在粮库、集贸市场及农户家中分两次采集样品 ,上半年采集贮存一年的小麦、玉米、大米各 30份 ,…     

食管癌高发区伏马菌素含量的检测

食管癌高发区伏马菌素含量的检测章红（中国农业大学生物学院，北京１０００９４）伏马菌素（Ｆｕｍｏｎｉｓｉｎｓ）是一类主要由串珠镰刀菌（Ｆｕｓａｒｉｕｍｍｏｎｉｌｉｆｏｒｍｅ）产生的真菌毒素，其中Ｆｕ－ｍｏｎｉｓｉｎＢ１（ＦＢ１）是天然污染玉米样品及真菌...     

部分克山病病区非病区稻谷玉米样品中串珠镰刀菌素含量的调查

为了探讨克山病的致病因素，利用薄层色谱（ＴＬＣ）和高效液相色谱（ＨＰＬＣ）法测定了四川、云南省克山病病区和北京市非病区稻谷中串珠镰刀菌素（ＭＦ）的含量，共计１２３份样品。病区８３份中，阳性率为８．８％，阳性样品中ＭＦ的含量范围为７３．５～２６５．３μｇ／ｋｇ；非病区４０份中，阳性率为２．５％，ＭＦ含量为１７９．５μｇ／ｋｇ。用ＨＰＬＣ法测定了黑龙江、吉林省克山病病区以及四川省、北京市非病区等地区玉米中的ＭＦ含量，共测定样品１０４份。病区４３份中，阳性率为８１．４％，阳性样品含量范围为５２．３～１１１６．０μｇ／ｋｇ；非病区６１份中，阳性率为９．７％，ＭＦ含量范围为７３．３～１０８２．１μｇ／ｋｇ。从以上测定结果分析，病区和非病区粮食样品中ＭＦ阳性率差异显著，但阳性样品的平均含毒水平差异无显著性；而不同粮食品种之间ＭＦ污染的差别远较不同地区（病区与非病区）之间的差别大。     

伏马菌素B1免疫检测方法的研究

本研究建立了用于检测粮食、饲料中伏马菌素Ｂ１（ＦＢ１）的快速、灵敏、简便的免疫检测方法—建立在单克隆抗体基础上的直接竞争性酶联免疫吸附测定方法（ＤＣ—ＥＬＩＳＡ法）;其最低检出浓度为１０ｎｇ／ｍｌ,线性范围在１０ｎｇ～５μｇ／ｍｌ。对新疆、安徽、黑龙江、哈尔滨四省市玉米样品中的ＦＢ１含量进行了测定,结果表明１５６份样品中有３４份检出ＦＢ１,阳性率为２１．７９％,含量在０．１８～３１．３２μｇ／ｇ范围内,平均含量为１２．０４μｇ／ｇ。     

串珠镰刀菌伏马菌素产毒基因及毒力的测定

目的研究我国串珠镰刀菌分离株伏马菌素产毒性与其生物合成酶基因fum5的关系。方法对不同省区、不同粮食样品中分离的29株串珠镰刀菌进行了产毒基因的测定,对其中18株产毒基因阳性株进行了伏马菌素生物合成,并用高效液相色谱法进行了毒素测定。结果26株为fum5基因阳性。除1株仅产生低水平的伏马菌素FB1外,其他菌株都产生不同水平的伏马菌素FB1、FB2和FB3,产毒量范围分别为0．41～140．20mg／kg、0．06～14．30mg／kg和0．30～58．00mg／kg。从芝麻中分离并鉴定了1株串珠镰刀菌伏马菌素强产毒株,分别产生高水平的伏马菌素FB1、FB2和FB3,产毒量分别达到128．84、11．80和14．88mg／kg。结论我国串珠镰刀菌分离株产生伏马菌素的能力与其生物合成酶基因rum5有密切关系。串珠镰刀菌可以污染芝麻并产生高水平的伏马菌素。     

串珠镰刀菌产伏马菌素和二相性真菌特性

目的 研究串珠镰刀菌产生伏马菌素和二相性真菌的致病性。方法 用间接竞争酶联吸附法测定从浙江省粮食中分离的 珠镰刀菌产竹的优良马菌素。结果 串珠镰刀菌产生的伏刀菌素Ｂ１〉４００μｇ／ｇ。１０份样品的串珠镰刀菌检出率在２￣１００％之间,伏马菌素Ｂ１污染量在３．７￣１４３．１μｇ／ｇ中珠刀菌在９％ＮａＣｌ沙保琼脂平板上３７℃培养可形成酵母样菌落,以出芽繁殖为主;在血平板上２可形成草绿色溶血反应。尚未见到     

胃癌、肝癌高发区和低发区粮食中杂色曲霉素污染量调查

采用间接竞争酶联免疫吸附试验（ＩＣ－ＥＬＩＳＡ）测定了２０８份粮食样品的杂色曲霉素（ＳＴ）污染量，发现其中７９份含ＳＴ少于４μｇ／ｋｇ，１１１份含４～３９μｇ／ｋｇ；ＳＴ污染量高于４０μｇ／ｋｇ的有１８份，最高含３４μｇ／ｋｇ。采自胃癌、肝癌高发区的１０７份样品，有４４．９％含２０μｇ／ｋｇ以上的ＳＴ，而低发区的６１份样品只有１４．８％含ＳＴ在２０μｇ／ｋｇ以上。二者，有极显著的差异（Ｐ＜０．０１）；高发区样品的ＳＴ污染量加权平均值为（１９．６±２１．６）μｇ／ｋｇ，低发区的为（１２．２±１１．８）μｇ／ｋｇ，二者也有极显著的差异（Ｐ＜０．０１）。在同一类地区，原粮的ＳＴ污染量显著高于成品粮的；不同粮食品种之间的ＳＴ污染量有差异，按ＳＴ污染量由大到小排列为：杂粮及饲料＞小麦＞稻谷＞玉米＞面粉＞大米。就单一品种比较，来自胃癌、肝癌高发区和低发区的小麦、玉米、大米三种粮食的ＳＴ污染量之间都有显著差异（Ｐ＜０．０５），而稻谷、面粉的ＳＴ污染量差异不显著（Ｐ＞０．０５）。对其中的１６份样品按现行国家标准检测方法──薄层层析法测定ＳＴ含量，发现１５份样品的测定结果与ＩＣ－ＥＬＩＳＡ法的测定值相一致。二种测定方法的符合?     

某食管癌高发区人群伏马菌素摄入量及尿二氢神经鞘氨醇/神经鞘氨醇比值的调查

为探讨伏马菌素与人类食管癌发生的关系 ,对食管癌高发区鹤壁市郊部分居民的伏马菌素摄入水平及尿二氢神经鞘氨醇 神经鞘氨醇 (Sa So)比值进行了调查。该地居民长期食用玉米 ,95 %的玉米样品检出了伏马菌素 ,其总伏马菌素含量 (伏马菌素B1、B2、B3之和 )为 7 2～ 72 6 8μg kg。估算受检者的伏马菌素摄入量为男性 0～ 4 84μg (kg·d) ,女性 0～ 4 0 4μg (kg·d)。检测他们的尿Sa So比值并与食管癌低发区睢县人群 (未接触伏马菌素 )比较 ,虽然两地女性尿Sa So比值间差异无显著性 ,但男性尿Sa So比值间差异存在显著性 ,食管癌高发区高于低发区。结果提示伏马菌素可能是食管癌发生的危险因素之一     

Toxicity of culture material of Fusarium verticillioides strain MRC 826 to nonhuman primates

We conducted a chronic feeding study in vervet monkeys (Cercopithecus aethiops) over 13.5 years. The experimental design consisted of two dietary treatment groups, each including males and females, fed varying levels of culture material of Fusarium verticillioides (Sacc.) Nirenberg (= F. moniliforme Sheldon) strain MRC 826 mixed into their daily food ration. Two females were included as treatment controls. We conducted blood chemical analyses bimonthly and recorded all clinical signs during the course of the experiment. We took liver biopsies at various stages during the initial phase of the experiment. Several monkeys were terminated in extremis during the experiment. Detailed feed intake profiles were determined 5 years after the experiment began, and the fumonisin B (FB) mycotoxin content of the feed was determined during the final stages of the experiment. The apparent FB consumption patterns were related to changes observed in the biochemical parameters in the blood and urine, including the liver function enzymes and creatinine clearance as well as differential blood counts and sphingolipid levels in the serum and urine. An apparent no-effect threshold for kidney and liver damage is estimated to be between 0.11 and 0.18 mg FB/kg body weight (bw)/day, which corresponds to a feed contamination level of between 8.21 and 13.25 mg FB/kg bw diet. Apart from the effects on the liver and kidney, a wide variety of parameters, including cholesterol and creatine kinase, were also adversely affected. Several blood parameters, including white and red blood cells, also significantly decreased in the treated animals. The serum sphinganine level and the sphingosine/sphinganine ratio, monitored toward the end of the experiment, significantly increased in both the low-dose and high-dose animals. The present study provides important information about the diversity of lesions induced by culture material of F. verticillioides in vervet monkeys and the dosage levels of fumonisins to be used in long-term studies in nonhuman primates.     

Discovery and occurrence of the fumonisins: a historical perspective

This article describes the events leading to the discovery of the fumonisins in South Africa in 1988 and highlights the first 10 years (1988-1998) of fumonisin research. The predominant fungus isolated from moldy corn implicated in a field outbreak of equine leukoencephalomalacia (ELEM) in South Africa in 1970 was Fusarium verticillioides (F. moniliforme). This fungus was also prevalent in moldy home-grown corn consumed by people in high-incidence areas of esophageal cancer (EC) in the Transkei region of South Africa. Culture material on corn of F. verticillioides strain MRC 826, which was isolated from moldy corn in Transkei, was shown to cause ELEM in horses, porcine pulmonary edema (PPE) syndrome in pigs, and liver cancer in rats. A short-term cancer initiation/promotion assay in rat liver was used to purify the carcinogen(s) in the culture material. These efforts finally met with success when fumonisins B1 and B2 novel mycotoxins with cancer-promoting activity in rat liver, were isolated from culture material of F. verticillioides MRC 826 at the Programme on Mycotoxins and Experimental Carcinogenesis of the Medical Research Council in Tygerberg, South Africa. Following the elucidation of the chemical structure of the fumonisins, these carcinogenic mycotoxins were shown to occur naturally in moldy corn in Transkei. Shortly thereafter, high levels of fumonisins in the 1989 U.S. corn crop resulted in large-scale field outbreaks of ELEM and PPE in horses and pigs, respectively, in the United States. Subsequently the fumonisins were found to occur naturally in corn worldwide, including corn consumed as the staple diet by people at high risk for EC in Transkei and China. These findings, together with the fact that the fumonisins cause field outbreaks of mycotoxicoses in animals, are carcinogenic in rats, and disrupt sphingolipid metabolism, have resulted in much worldwide interest in these compounds during the first 10 years after the discovery of the fumonisins in 1988.     

串珠镰刀菌伏马菌素产毒株聚合酶链反应检测方法的研究

以伏马菌素生物合成所必需的多酮肽合成酶基因fum5为基础 ,设计了 3对特异性引物P1 P2、P3 P4和Fum5F2 1 Fum5 1,建立了串珠镰刀菌伏马菌素产毒株PCR检测方法。以 5株ATCC典型串珠镰刀菌及其他菌株为参考 ,其中ATCC 5 2 5 3 9为伏马菌素B1(FB1)产毒株 ,测定了PCR方法的特异性。ATCC 5 2 5 3 9扩增结果阳性 ,该 3对引物分别扩增出 880bp、70 2bp和 10 40bpDNA片段 ,非伏马菌素产毒株ATCC 3 8946、ATCC2 62 63、ATCC 12 763、ATCC 3 80 16及其他阴性对照菌株 (禾谷镰刀菌、梨孢镰刀菌、木贼镰刀菌、三线镰刀菌、黄曲霉、拟青霉和大肠杆菌O15 7)结果阴性 ,证明引物具有很好的特异性。同时以不同稀释度的ATCC 5 2 5 3 9DNA为模板 ,测定了 3对引物PCR的敏感性 ,P1 P2每PCR反应的检出限为 10 0pg ,P3 P4和Fum5F2 1 Fum5R1每PCR反应的检出限均为 10pg ,分别相当于每PCR反应检出 10 4和 10 3个孢子。 3对引物PCR检测国内不同地区分离的 3 2株串珠镰刀菌及交孢变种 ,2 6株为伏马菌素产毒株 ,6株为非伏马菌素产毒株。并对部分产毒株PCR(P3 P4)产物 ,应用内切酶EcoRV和BamHI,进行限制片段长度多态性 (RFLP)分析 ,分别产生 181bp、5 2 1bp 2个和 116bp、2 5 8bp、3 2 8bp 3个DNA片段 ,与文献报道值一致 ,确证了结     

Fumonisin toxicosis in swine: an overview of porcine pulmonary edema and current perspectives

Fumonisin toxicosis in swine was named porcine pulmonary edema (PPE) after outbreaks of a fatal disease in pigs fed Fusarium verticillioides (F. moniliforme)-contaminated corn screenings from the 1989 corn crop in Iowa, Illinois, and Georgia. Pigs that died had severe pulmonary edema, which has not been identified in other species after exposure to fumonisins. The disease has been reproduced experimentally by feeding of naturally contaminated corn, F. verticillioides culture material, and by intravenous administration of fumonisin B1 (FB1). Hepatic lesions consisting of apoptosis, necrosis, and hepatocyte proliferation also are observed. As in other species, alterations in clinical pathology reflect hepatic injury as well as elevated serum cholesterol concentration. In chronic studies, esophageal plaques, hyperplastic hepatic nodules, and right ventricular hypertrophy were found. In pigs, as in other species, fumonisin alters sphingolipid biosynthesis, with the greatest alterations in sphingosine and sphinganine concentrations in kidney, liver, lung, and heart. Our recent studies on fumonisin toxicosis in pigs have focused on immune effects and the pathogenesis of pulmonary edema. The specific immune system was not affected; however, FB1 inhibited phagocytosis and sphingolipid biosynthesis in pulmonary macrophages. Fumonisin induced an accumulation of membranous material in pulmonary capillary endothelial cells; this change appears specific to this cell type and to swine. In short-term cardiovascular studies, fumonisin decreased left ventricular dP/dt(max) (an index of cardiac contractility), mean systemic arterial pressure, heart rate, and cardiac output, and increased mean pulmonary artery pressure and pulmonary artery wedge pressure. These changes are compatible with the inhibition of L-type calcium channels by increased sphingosine and/or sphinganine concentration. Therefore, fumonisin-induced pulmonary edema in swine appears to result from acute left-sided heart failure mediated by altered sphingolipid biosynthesis.     

Increases in serum sphingosine and sphinganine and decreases in complex sphingolipids in ponies given feed containing fumonisins, mycotoxins produced by Fusarium moniliforme.

Consumption of food contaminated with Fusarium moniliforme causes leucoencephalomalacia and hepatotoxicity in horses, pulmonary edema in pigs and liver cancer in rats, and has been correlated with esophageal cancer in humans. The causative agents are thought to be a family of compounds called fumonisins, which have recently been shown to be potent inhibitors of sphingosine (sphinganine) N-acyltransferase. Because inhibition at this step blocks the formation of complex sphingolipids while leading to accumulation of sphinganine, we hypothesized that exposure of animals to fumonisin-contaminated feed might be detected by analyses of serum sphingolipids. Within days of giving ponies feed contaminated with 15 to 44 micrograms/g fumonisin B1, there was an increase in the amount of free sphinganine (and sometimes sphingosine) and a reduction in complex sphingolipids. Free sphinganine and sphingosine decreased when ponies consumed less of the contaminated feed, and increased again when they consumed more fumonisin. When toxicosis was evident as indicated by other serum markers, complex sphingolipids as well as free sphingosine and sphinganine were elevated, probably due to loss of sphingolipids from dying cells. These findings establish that consumption of fumonisin-contaminated feed disrupts sphingolipid metabolism. Because the changes in sphinganine and sphingosine were seen before liver enzymes were noticeably elevated, they may be an early marker of exposure to fumonisins.