贝莱斯芽孢杆菌DJB5的生物安全性评价  

Biosafety evaluation of Bacillus velezensis DJB5

作　　者：唐萍[1] 代飞燕 吴毅歆[3,4] 叶敏 何月秋[3,4] TANG Ping;DAI Fei-yan;WU Yi-xin;YE Min;HE Yue-qiu(College of Plant Protection,Yunnan Agricultural University,Kunming 650201,China;College of Animal Medicine,Yunnan Agricultural University,Kunming 650201,China;College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China;National and Local Joint Engineering Center for Screening and Application of Microbial Strains,Kunming 650217,China)

机构地区：[1]云南农业大学植物保护学院,昆明650201 [2]云南农业大学动物医学院,昆明650201 [3]云南农业大学农学与生物技术学院,昆明650201 [4]微生物菌种筛选与应用国家地方联合工程研究中心,昆明650217

出　　处：《南方农业学报》2019年第12期2720-2727,共8页Journal of Southern Agriculture

基　　金：云南省现代农业玉米产业技术体系建设专项(2017KJTX002);云南省教育厅科学研究基金项目(2018JS267)

摘　　要：【目的】研究贝莱斯芽孢杆菌DJB5的生物安全性,为DJB5的开发和安全使用打下基础。【方法】参考微生物菌剂安全性的相关标准,通过灌胃DJB5菌体和发酵液的方式测定DJB5对昆明小鼠的急性经口毒性;通过腹腔注射菌体的方法研究DJB5的致病性;测试DJB5对昆明小鼠的急性眼刺激性、皮肤刺激性和溶血性;用纸片琼脂扩散法检测DJB5的抗菌药物敏感性。【结果】DJB5菌体对昆明小鼠的急性经口半致死剂量(LD50)>5000 mg/kg,灌胃菌体(5000 mg/kg BW)及5和10倍发酵上清浓缩液14 d后,昆明小鼠的体征、各脏器石蜡切片、血相和生化指标与对照组无显著差异(P>0.05,下同);腹腔注射DJB5菌体(500 mg/kg BW)后,处理组昆明小鼠的脏器系数和体重与对照组无显著差异;DJB5对昆明小鼠眼和皮肤无刺激性、无溶血性,且对受试的青霉素G、头孢唑林、丁胺卡那、庆大霉素、红霉素、诺氟沙星、环丙沙星、复方新诺明和氯霉素等9种抗菌药物敏感。【结论】贝莱斯芽孢杆菌DJB5及其发酵液的生物安全性高,具有开发成防治玉米贮藏期黄曲霉制剂的潜力。【Objective】The biosafety of Bacillus velezensis DJB5 was studied to lay a foundation for the development and safe application of DJB5.【Method】Based on standards of microbial agent safety,the acute oral toxicity was studied with stomach perfusion of DJB5 bacterial cells and fermentation supernatant,and pathogenicity was determined by the ways of intraperitoneal injections. Acute eyes,skin irritations and hemolytic tests were proceeded in Kunming mice. Antibiotic resistance was also tested by agar diffusion method.【Result】The results displayed that its acute oral toxicity median lethal dose(LD50)was >5000 DJB5 cells/kg,stomach filling with maximum 5000 mg/kg BW DJB5 and 5 and 10 times of concentrated formulated suspension for 14 d did not cause significant differences of body sign,organs section,hemogram biochemical indicators with negative control(P>0.05,the same below). Intraperitoneal injection with 5000 mg/kg BW DJB5 did not cause any difference of Kunming mouse’s organ coefficient and weight with negative control. Strain DJB5 did not irritate Kunming mouse’s eyes and skin. It did not show hemolysis,but was sensitive to nine antibiotics including penicillin G,cefazolin,amikacin,gentamicin,erythrocin,norfloxacin,ciprofloxacin,selectrin,and chloramphenicol.【Conclusion】Results indicates that strain DJB5 and its formulated suspension are biologically safe and has the potential to develop a biocontrol agent to control Aspergillus flavus.

关 键 词：贝莱斯芽孢杆菌DJB5 生物安全性 抗生素 黄曲霉 

分 类 号：S182[农业科学—农业基础科学]