犬支气管败血波氏杆菌的分离鉴定和灭活疫苗初步应用  

Isolation and Identification of Bordetella bronchiseptica in Canines and Preliminary Application of Inactivated Vaccine

作　　者：王潘龙 赵悦 孙强 周昊 尹月 曲雪婷 原昆鹏 栾庆东 陈淑贞 卢艳敬 臧立玉 韩海峰 尹燕博[1,3] WANG Panlong;ZHAO Yue;SUN Qiang;ZHOU Hao;YIN Yue;QU Xueting;YUAN Kunpeng;LUAN Qingdong;CHEN Shuzhen;LU Yanjing;ZANG Liyu;HAN Haifeng;YIN Yanbo(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China;Qingdao Jimo District Animal Husbandry Development Service Center,Qingdao 266299,China;Qingdao Bolong Experimental Animal Co.,Ltd.,Qingdao 266225,China;College of Veterinary Medicine,Shanxi Agricultural University,Jinzhong 030801,China;College of Veterinary Medicine,Dongbei Agricultural University,Haerbin 150030,China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109 [2]青岛市即墨区畜牧业发展服务中心,山东青岛266299 [3]青岛博隆实验动物有限公司,山东青岛266225 [4]山西农业大学动物医学院,山西晋中030801 [5]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国兽医杂志》2024年第7期41-48,共8页Chinese Journal of Veterinary Medicine

基　　金：青岛市新兴产业培育计划项目(23-1-4-xxgg-21-nsh)。

摘　　要：某实验比格犬养殖场部分犬出现咳喘和口鼻流血等症状,严重者死亡。为了调查发病原因并采取针对性措施,本试验采集临床发病犬的鼻腔拭子、死亡犬的肺脏和气管分泌物等,进行细菌分离培养;通过形态观察、革兰染色、16S rRNA和ptxA基因测序,对分离细菌进行鉴定,并对部分菌株进行药敏试验。选取1株致病性强的分离菌株制备支气管败血波氏杆菌蜂胶佐剂灭活疫苗,进行免疫保护试验,分别采用1.0和0.5 mL剂量免疫小鼠,免疫后21 d攻毒;对养殖场临床发病犬和同群健康犬进行紧急免疫接种,剂量为4月龄以上犬每只1.0 mL、2~4月龄犬每只0.5 mL。对临床病死犬和攻毒死亡小鼠进行组织病理学观察。结果显示,从临床发病犬的鼻腔拭子、肺脏、气管和鼻腔黏液状分泌物共分离鉴定获得30株支气管败血波氏杆菌。药敏试验结果显示,临床分离菌株对环丙沙星和米诺环素高度敏感,但临床治疗效果不明显。免疫保护试验中,1.0 mL和0.5 mL蜂胶佐剂灭活疫苗对小鼠的保护率均可达100%;紧急免疫接种后,临床发病犬逐渐恢复,同群健康犬无新发病个体,疫苗免疫效果良好。组织病理学观察发现,临床病死犬和攻毒死亡小鼠均表现典型的支气管肺炎和肺气肿病变,肝细胞呈水泡样变。结果表明,支气管败血波氏杆菌是导致该场部分犬出现呼吸道疾病的病原体,蜂胶灭活疫苗可有效防控实验用比格犬支气管败血波氏杆菌感染。Some Beagle dogs at a breeding farm exhibited symptoms of coughing,wheezing,and bleeding from the mouth and nose,with severe cases resulting in death.To investigate the cause and implement targeted measures,clinical samples including nasal swabs from affected dogs,lung tissue and tracheal secretions from affected dogs were collected for bacterial isolation and culture.The isolated bacteria were identified through morphological observation,Gram staining,and sequencing of the 16S rRNA and ptxA genes.Antibiotic sensitivity tests were conducted on selected strains.A virulent strain was used to prepare a Bordetella bronchiseptica inactivated vaccine with a propolis adjuvant,which was tested for immunoprotection in mice using doses of 1.0 mL and 0.5 mL,followed by challenge infection 21 days post-immunization.Emergency vaccinations were administered to clinically affected and healthy dogs in the breeding farm,with doses of 1.0 mL for dogs over 4 months old and 0.5 mL for dogs aged 2-4 months.Histopathological examinations were conducted on tissues from clinically deceased dogs and challenge deceased mice.The results showed that 30 strains ofBordetella bronchisepticawere isolated from nasal swabs,lung tissue,trachea,and nasal secretions of affected dogs.Antibiotic sensitivity tests revealed high susceptibility of clinical isolates to ciprofloxacin and minocycline,though clinical treatment was not markedly effective.In the immunoprotection test,the propolis-adjuvanted inactivated vaccine provided 100% protection in mice at both 1.0 mL and 0.5 mL doses.Following emergency vaccination,clinically affected dogs gradually recovered,and no new cases were observed among the healthy cohort,indicating effective vaccine protection.Histopathological findings in clinically deceased dogs and challenge deceased mice showed characteristic bronchopneumonia,pulmonary emphysema,and vacuolar degeneration of hepatocytes.The results indicate thatBordetella bronchisepticawas the causative agent of the respiratory disease outbreak in the breed

关 键 词：比格犬 支气管败血波氏杆菌 16S rRNA 组织病理学观察 蜂胶佐剂灭活疫苗 

分 类 号：S855.3[农业科学—临床兽医学]