鸭疫里默氏杆菌血清2型的分离鉴定及商品化疫苗免疫保护效果研究

doi:10.16431/j.cnki.1671-7236.2025.09.025

摘要/Abstract

摘要： 【目的】本研究拟对从发病雏鸭中分离获得的鸭疫里默氏杆菌(Riemerella anatipestifer，RA)进行生物学特性研究，并通过动物攻毒试验评估当前市售的3种鸭传染性浆膜炎商品化疫苗对新分离菌株的保护效果，以期为鸭传染性浆膜炎的防控提供参考。【方法】采集病死雏鸭肝脏、脑组织分离病原菌，通过形态学观察、16S rRNA基因PCR鉴定和测序、系统进化树构建、血清型分型等方法对分离菌株进行鉴定。通过分光光度法和平板计数法测定分离菌株生长曲线，利用纸片扩散法分析其药物敏感性，通过动物试验测定其半数致死量(LD₅₀)和组织载菌量，最后通过攻毒试验评估3种商品化灭活疫苗对分离菌株的免疫保护效果。【结果】试验分离获得1株RA，其为血清2型。药敏试验结果显示，分离株对氨苄西林、头孢他啶和卡那霉素等10种药物均表现耐药,对阿莫西林、头孢噻肟、氟苯尼考和链霉素表现敏感。生长曲线测定结果显示，培养6～10 h，分离株进入对数生长期，培养8 h后活菌数可达到10⁹ CFU/mL；对雏鸭的LD₅₀为7.65×10^4.7 CFU，雏鸭感染后心脏组织载菌量最高。3种商品化灭活疫苗免疫7日龄雏鸭后2周用分离菌株进行攻毒，结果显示，1+2+7型多价RA灭活疫苗保护效果最好，保护率可达70%，而另外2种疫苗保护率＜50%。【结论】本研究分离获得1株多重耐药、高致病性血清2型RA,当前市售RA商品化疫苗对其免疫保护效果有限。研究结果为当前鸭传染性浆膜炎的防控提供理论依据。

关键词: 鸭疫里默氏杆菌; 血清2型; 生物学特性; 商品化疫苗

Abstract: 【Objective】 The aim of this study was to investigate the biological characteristics of Riemerella anatipestifer (RA),which was isolated from diseased ducks,and evaluate the protective effect of 3 commercially available vaccines for infectious serositis in ducks against the newly isolated strain through animal challenge tests,in order to provide a reference for the prevention and control of infectious serositis in ducks. 【Method】 The liver and brain tissues of diseased ducklings were collected to isolate pathogenic bacteria.The isolated strains were identified by morphological observation,16S rRNA gene PCR identification and sequencing,phylogenetic tree construction,and serotype typing.The growth curves of the isolated strains were determined by spectrophotometry and plate counting.The drug sensitivity was analyzed by disk diffusion method.The median lethal dose (LD₅₀) and tissue bacterial load were determined through animal experiments.Finally,the immune protective effects of 3 commercial inactivated vaccines on the isolated strains were evaluated through challenge tests. 【Result】 One strain of RA was isolated in this experiment,and it was serotype 2.The results of the drug sensitivity test showed that the isolate was resistant to 10 drugs such as ampicillin,ceftazidime and kanamycin,and was sensitive to amoxicillin,cefotaxime,florfenicol and streptomycin.The determination results of the growth curve showed that after 6-10 h of culture,the isolate entered the logarithmic growth phase,and the viable bacteria count could reach 10⁹ CFU/mL after 8 h of culture.The LD₅₀ for ducklings was 7.65×10^4.7 CFU,and the bacterial load in heart of ducklings was the highest after infection.2 weeks after immunizing 7-day-old duckings with 3 commercial inactivated vaccines,the isolate was used for challenge.The results showed that the multivalent RA inactivated vaccines of types 1+2+7 had the best protective effect,with a protection rate of up to 70%,while the protection rates of the other two vaccines were less than 50%. 【Conclusion】 In this study,one strain of multi-drug resistant and highly pathogenic RA serotype 2 was isolated，the immune protective effect of commercially available RA vaccines on the isolated was limited.This result provided a theoretical basis for the current prevention and control of infectious serositis in ducks.

Key words: Riemerella anatipestifer; serotype 2; biological characteristics; commercial vaccine

中图分类号:

S852.61

张蓉蓉, 汪最, 郭云清, 李国红, 程伊洛, 卢琴, 罗聪, 张腾飞, 张文婷, 胡巧, 罗青平. 鸭疫里默氏杆菌血清2型的分离鉴定及商品化疫苗免疫保护效果研究[J]. 中国畜牧兽医, 2025, 52(9): 4273-4283.

ZHANG Rongrong, WANG Zui, GUO Yunqing, LI Guohong, CHENG Yiluo, LU Qin, LUO Cong, ZHANG Tengfei, ZHANG Wenting, HU Qiao, LUO Qinping. Isolation and Identification of Riemerella anatipestifer Serotype 2 and the Evalution of Protective Efficacy of Commercial Vaccine[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(9): 4273-4283.

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