禽致病性大肠杆菌lpxL和lpxM缺失减毒活疫苗的安全性和保护效果评价  

Evaluation of Efficacy and Safety of the Live Attenuated Vaccine Based on Avian Pathogenic Echerichia coli with Deficient lpxL and lpxM

作　　者：蒋路遥 祝谢民 吴佳炎 杨帆 潘昊 黄恩赐 高清清 郇长超 王小波 高崧 JIANG Luyao;ZHU Xiemin;WU Jiayan;YANG Fan;PAN Hao;HUANG Enci;GAO Qingqing;HUAN Changchao;WANG Xiaobo;GAO Song(College of Veterinary Medicine(Institute of Comparative Medicine),Yangzhou University,Yangzhou,Jiangsu 225009;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou,Jiangsu 225009;Key Laboratory of Avian Bioproduct Development,Ministry of Agriculture and Rural Affairs,Yangzhou,Jiangsu 225009;Institutes of Agricultural Science and Technology Development,Yangzhou University,Yangzhou,Jiangsu 225009)

机构地区：[1]扬州大学兽医学院(比较医学研究院),江苏扬州225009 [2]江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [3]农业农村部禽用生物制剂创制重点实验室,江苏扬州225009 [4]扬州大学农业科技发展研究院,江苏扬州225009

出　　处：《中国家禽》2025年第5期62-70,共9页China Poultry

基　　金：国家自然科学基金项目(32373011);国家重点研发计划(2017YFD0500705);江苏高校优势学科建设工程资助项目(PAPD)。

摘　　要：为研究禽致病性大肠杆菌(Avian pathogenic Escherichia coli,APEC)lpxL和lpxM缺失减毒活疫苗对SPF鸡的安全性和免疫保护效果,试验通过对大肠杆菌双基因缺失减毒活疫苗株体外遗传稳定性、疫苗培养工艺、免疫方式优化和免疫攻毒保护效果评价,探讨其作为禽大肠杆菌病活疫苗候选株的可能性。结果显示:双基因缺失减毒活疫苗株E516ΔlpxLΔlpxM、E058ΔlpxLΔlpxM和E522ΔlpxLΔlpxM在体外传代50代遗传稳定,未发生回复突变,毒力较野生株下降显著(10000倍以上),优化的培养参数为液体初始培养12 h,以终浓度OD_(600 nm)为0.15转至LB肉汤再培养5 h,此时活菌比最高,优化的免疫方式和剂量为以1×10^(9)CFU/只(0.2 mL)进行颈部皮下接种具有良好的保护效果。研究表明,lpxL和lpxM缺失的APEC双基因缺失减毒活疫苗株具有禽大肠杆菌病活疫苗候选株的潜能。This study aimed to investigate the safety and immune efficacy of avian pathogenic Escherichia coli(APEC)lpxL and lpxM deletion attenuated live vaccines in specific pathogen free(SPF)chickens.The experiment evaluated the genetic stability of the double-gene-deleted vaccine strains in vitro,vaccine strain cultivation,immunization methods and the efficacy of prototype vaccine in chickens against APEC challenge.The results showed that the double-gene-deleted attenuated vaccine strains E516ΔlpxLΔlpxM,E058ΔlpxLΔlpxM and E522ΔlpxLΔlpxM maintained genetic stability over 50 generations in vitro without gene reversion,and their virulence was significantly reduced compared to the wild-type strains(more than 10000-fold).The optimized cultivation parameters involved initial liquid culture for 12 h,followed by inoculation into LB broth at a final concentration of OD_(600 nm)=0.15 for an additional 5 h,at which point the highest viable cell ratio was achieved.The optimized immunization method and dosage involved subcutaneous injection at the neck with 1×10^(9) CFU per bird(0.2 mL),which provided sound protective effects.The results indicated that APEC attenuated vaccine strains with deletions in lpxL and lpxM had the potential to serve as candidate strains for APEC gene-deleted attenuated live vaccines.

关 键 词：禽致病性大肠杆菌 基因缺失 活疫苗 lpxL lpxM 

分 类 号：S855.1[农业科学—临床兽医学]