【作者】 王海燕；

【导师】 牟志美；

【作者基本信息】 山东农业大学 ， 特种经济动物饲养， 2002， 硕士

【摘要】 家蚕(Bombyx mori)细菌性中毒症是家蚕食下苏云金芽孢杆菌(Bacillusthuringiensis，简称B.t)所产生的芽孢和伴孢晶体而引起的急性中毒症，又称卒倒病。该病原广泛存在于各种土壤及空气中，具有很强的杀虫活性，至少能杀死节肢动物门九个目及原生动物门、线形动物门和扁形动物门中的多种动物，而对人、畜、哺乳动物及天敌等无害，因此，B.t是生防工业中应用最广泛的微生物。而随着BT制剂的广泛应用，家蚕卒倒病越来越严重地威胁着我国的蚕业生产。因此，研制性状稳定、对该菌有强烈杀灭效果的消毒药剂对减少蚕病的发生具有重要意义，而其作用机理的研究又能为生产实践提供理论依据。本文应用生物鉴定技术、电泳技术、电镜技术等从生化指标分析及超微形态观察等方面研究了对B.t有高效灭活作用的氯制剂对B.t伴孢晶体及芽孢的灭活机理。研制出一种以二氯异氰尿酸钠为主剂，加以具有增效、缓蚀、渗透、扩散等作用的辅剂而成的一种一剂多用途的复方消毒剂XDJ-Ⅲ。主要取得以下研究结果： 1．SDS—聚丙烯酰胺凝胶电泳结果表明，经优氯净处理的伴孢晶体的电泳谱带发生了明显的变化，说明氯制剂破坏了伴孢晶体的一级结构。 2．样品的扫描电镜观察表明，氯制剂能够破坏伴孢晶体的表面结构，使伴孢晶体表面出现破损，进而出现孔洞，较长时间作用下可以使伴孢晶体完全失去原有外貌呈不规则的蓬松团状，即立体结构受到破坏。伴孢晶体的一级结构和立体结构被破坏导致伴孢晶体的毒力降低甚至完全丧失。 3．氯制剂能够破坏芽孢的表面结构。使芽孢外壁结构失去致密性，从而减弱了其保护作用。 4．氯制剂能够破坏芽孢的内部结构。芽孢外壁被破坏以后，药液渗透进入芽孢内部，溶解破坏内部组织。破坏到一定程度则芽孢的萌发力降低甚至丧失。 5．以具有高度稳定性和安全性的二氯异氰尿酸钠为主剂，配以几种具有增效、缓蚀、渗透、扩散等作用的辅剂研制而成的复方消毒剂XDJ-Ⅲ可以配成溶液用于蚕室、蚕具及叶面消毒，对家蚕的各种病原均具有较强的杀灭作用。也可以直接点燃用于蚕室蚕具和蚕座的熏烟消毒，作为熏烟剂发烟迅速、无明火，有效成分释放率为99.8％以上，残留低。 6．XDJ-Ⅲ用于蚕室蚕具消毒使用有效氯浓度为0.5％。叶面消毒以有效氯浓度为0.3％为宜，要现配现用。蚕室蚕具熏烟消毒以5～8g／m~3，注意密闭门窗12hr以上。养蚕中蚕体蚕座消毒以1.5g／m~3，30min后通风换气，对家蚕的生长发育无不良影响。 I 王海燕：优氯净对苏云金杆菌灭活机理及复方消毒剂的研究 7．XDJ－Il对金属的缓蚀作用明显；生产工艺简单，储存使用方便安全，能很好地满足蚕业生产上对多用途消毒剂的要求。更多还原

【Abstract】 si\kwoTm(Bombyx mori) bacterial toxicosis was caused by Bacillus thuringiensis (B.t) when silkworm ate enough amount of its spore and parasporial crystal .It was an acute toxicosis and its pathogeny was Bacillus thuringiensis .It lied in all kinds of soil and air. B.t had strong insectcidal ability ,and it could kill a lot of creatures of Arthropoda, Protozoa, Nemathelminthes and Platyhelminthes.B.t was the most comprehensive microorganism being used in biotic pesticide.With the more comprehensive use of BT biotic pesticidal, silkworm bacterial toxicosis threntened the development and safety of sericulture in our country.For these reasons, it had great significance developing new medicine with steady character and excellent disinfectant effect to B.t to prevent silkworm from infecting this disease.And the study of disinfectant mechanism of chlorine preparation to parasporal crystal and spore of Bacillus thuringiensis could provide practice with academic direction.We studied these disinfectant mechanisms of sodium dichloroisocyanurate to parasporal crystal and spore by biological assay,electron microscope technique,electrophoresis technique and so on.The major achievements obtained in these experiments were as follows.1. The result of SDS-PAGE indicated that the pedigrees of the treated parasporal crystal had changed obviously and chlorine preparation distroyed primary structure of parasporal crystal and made it lose its activity.2. The result of SEM showed that sodium dichloroisocyanurate damaged the shape structure of parasporal crystal, and then holes appeared on its surface.If treatment time was long enough,the parasporal crystal would lose its former41appearance and became loose corps,that is,its stereoscopic structure was damaged.The damage of primary and stereoscopic structure of parasporal crystal made its virulence receded and even lost completely.3. chlorine preparation could destroy external structure of spore. And the fabric of spore was no longer compact,thus its ability of protection was lower.4. chlorine preparation could destroy inner structure of spore. The medicine penetrating into spore damaged inner tissue.When the damage was serious enough,the germination ability of spore was lower and even lost completely.5. We developed a compound disinfectant ,named XDJ-III, using sodium dichloroisocyanurate as main medicament which had high stability and security and some substance as accessorial medicament which could increase effect,postpone erosion, increase penetration ability and pervasion ability. XDJ-III could disinfect silkworm rooms and tools .leaves surface when being confected to a certain concentration solution. It had good disinfecting effect on silkworm’s main pathogens.At the same time,it could be used as smoking medicine disinfecting silkworm rooms.Being used as smoking medicine, XDJ- El had many strongpoints.For example,it could give out smoke rapidly and had no eyeable fire.lt could loose its active ingredient more than 99.8% and the leftover was very little.6. Being used as disinfectant of silkworm rooms and tools, the available chlorine concentration of XDJ-III should be 0.5%. As disinfecting leaves, the available chlorine concentration of XDJ-III should be 0.3%. We’d better use up the solution, otherwise it would decrease its effect. Being used as smoking disinfectant, its effective concentration was about 5~8g/m3,and we must close windows and doors more than 12 hours. And when disinfecting silkworm and rearing bed, its concentration was about 1.5g/m3 and 30 minutes later we should air the silkworm room. It did not have bad influence on silkworm growth and development.7. XDJ-III had obvious effect of postponing erosion of metal. The manufacture technics , preservasion and use were very simple and easy .It could meet the need of multiuse medicine?in sericulture.更多还原