癌变·畸变·突变

• 论著 • 上一篇    下一篇

应用PCR法检测微生物类农药木霉菌可湿性粉剂急性经口致病性

LIU Ying-hua,ZHANG Jing-shu*,JIANG Shu-qing,HE Ning   

  1. Tianjin Center for Disease Control and Prevention, Tianjin 300011, China
  • 收稿日期:2012-08-07 修回日期:2012-09-25 出版日期:2011-11-30 发布日期:2011-11-30
  • 通讯作者: 张静姝,Tel,022-24333606,E-mail:rainbow 7546@ 163.com
  • 作者简介:刘英华 (1977- ),女,山东省济宁市人,硕士,主管技师,Tel: 022-24333605, E-mail:liuyinghua36@163.com

PCR detection of acute oral pathogenicity of microbial pesticide trichoderma wettable powder

天津市疾病预防控制中心,天津 300011   

  1. Tianjin Center for Disease Control and Prevention, Tianjin 300011, China
  • Received:2012-08-07 Revised:2012-09-25 Online:2011-11-30 Published:2011-11-30
  • Contact: ZHANG Jing-shu,Tel,022-24333606,E-mail:rainbow 7546@ 163.com
  • About author:刘英华 (1977- ),女,山东省济宁市人,硕士,主管技师,Tel: 022-24333605, E-mail:liuyinghua36@163.com

摘要:

目的: 应用PCR法检测微生物类农药木霉菌可湿性粉剂的急性经口致病性,探讨PCR法在微生物类农药经口致病性研究中的应用。方法:Wistar大鼠共56只,分成对照组和染毒组,每组28只,雌雄各半。选用微生物农药木霉菌可湿性粉剂为受试样品,染毒组每只大鼠按108个孢子经口灌胃,对照组灌胃等量生理盐水。于染毒后第1、7、14、21天收集动物粪便,同时剖解动物的肾脏、心脏和肝脏等组织提取DNA,利用木霉菌特异性引物进行PCR扩增,检测受试物目的基因的表达,以观察受试微生物农药的感染性、持久性。并记录观察期内动物的中毒症状、死亡情况、体质量,对死亡动物进行大体解剖并进行病理组织学检查,以判断其致病性。结果:染毒后仅从第1天染毒大鼠的粪便中提取的DNA,经PCR检测可见木霉菌目的基因扩增条带,染毒后第7天及以后粪便中均未见目的基因表达,其余各脏器提取的DNA,经PCR扩增后亦均未见目的基因表达。动物染毒后,未见明显中毒表现,观察期内无动物死亡,大体检查及病理检查均未见明显异常。结论:木霉菌可湿性粉剂经口灌胃进入动物体内后仅在胃肠道内短期存在,并逐渐被清除掉,并未透过胃和肠道而侵染进入动物的组织、器官中。微生物农药木霉菌可湿性粉经口染毒后对大鼠无致病性。应用PCR可有效检测出受试物的清除情况,且具有较好的特异性,可作为微生物农药经口致病性的检测方法。

关键词: 微生物农药, 急性经口毒性, 致病性, 木霉菌

Abstract:

OBJECTIVE: To detect the acute oral toxicity/pathogenicity of trichoderma wettable powder (WP),and to explore the application of PCR method in the pathogenicity of microbial pesticides.  METHODS:56 Wistar rats were randomly divided into a control group and exposed groups with 28 rats in each group and half male and half female. Trichoderma WP was selected as the test sample. The rats of exposure group were treated with 108 spores by gavage. Animals of control group were gavaged with normal saline. At the 1st,7th,14th and 21st day after exposure,stools were collected,then rats were sacrificed and the kidneys,liver,heart,and other organs were eviscerated. DNA of the tissue samples were extracted,then amplified with PCR. The symptoms of poisoning including mortality and body weight of animals were recorded. Dead animals were dissected and histopathological examination were performed. RESULTS: Only the amplification products of stools on the 1st day appeared positive but not on the 7th,14th and 21st days.  The amplification products of the remaining organs did not appear positive. No mortality was observed after exposure . In the histopathological examination no pathological changes were found. CONCLUSION: The trichoderma was found to exist only in the gastrointestinal tract,and was gradually excreted,It did not infect the animal’s tissues and organs,suggesting the test sample had no pathogenicity in rats.  PCR method could effectively detect the excretion of the test substance,with a good specificity which could be used as a detection method for oral toxicity/pathogenicity of microbial pesticide.

Key words: microbial pesticides, acute oral toxicity, pathogenicity, trichoderma