猪链球菌2型制苗用菌株的筛选

doi:10.3969/j.issn.1004-1524.2020.01.07

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (1): 57-64.DOI: 10.3969/j.issn.1004-1524.2020.01.07

猪链球菌2型制苗用菌株的筛选

陈章¹, 吴华健¹, 毛天骄¹, 韩业芹¹, 孙裴¹, 魏建忠¹, 李东风², 李郁^1,*

1. 安徽农业大学动物科技学院,安徽合肥 230036;
2. 安徽省兽药饲料监察所,安徽合肥 230001

收稿日期:2018-12-16 出版日期:2020-01-25 发布日期:2020-03-11
通讯作者: ^*李郁,E-mail:liyouer@163.com
作者简介:陈章(1995—),男,安徽合肥人,硕士研究生,研究方向为动物传染病学。E-mail:554323097@qq.com
基金资助:
国家星火计划重点项目(2014GA710002); 安徽省长三角联合科技攻关项目(1101c0603065); 安徽省质量工程项目(2013sxzx008); 安徽省生猪产业体系基金(皖农科〔2016〕84号)

Screening of strain for inactivated vaccine of Streptococcus suis type 2

CHEN Zhang¹, WU Huajian¹, MAO Tianjiao¹, HAN Yeqin¹, SUN Pei¹, WEI Jianzhong¹, LI Dongfeng², LI Yu^1,*

1. College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;
2. Veterinary Drug and Feed Surveillance Institute of Anhui Province, Hefei 230001, China

Received:2018-12-16 Online:2020-01-25 Published:2020-03-11

摘要/Abstract

摘要： 为筛选出致病力强、抗原性优良、遗传性状相对稳定的猪链球菌2型(Streptococcus suis 2,SS2)制苗用菌株。以6株临床分离SS2强毒株为受试菌株(编号为HF1、XC1、HF2、HF3、BZ1、BB1),通过改良寇氏法测定菌株半数致死量(LD₅₀),利用ELISA检测新西兰大白兔和昆明鼠血清的抗体效价测定反应原性,昆明鼠及斑马鱼免疫攻毒试验测定免疫原性,同时连续传代培养受试菌株,检测第10代、20代和30代菌株的半数致死量(LD₅₀)、血清抗体效价和免疫保护率。结果显示,BB1、BZ1、XC1、HF1、HF2和HF3对昆明鼠的LD₅₀分别为3.72×10⁹、3.31×10⁸、1.58×10⁹、1.00×10⁹、5.01×10⁸和3.24×10⁸ CFU·mL^-1;对斑马鱼的LD₅₀分别为3.31×10³、0.93×10²、6.03×10³、3.39×10⁴、0.62×10²和2.34×10³ CFU·mL^-1。ELISA抗体效价测定和免疫攻毒试验结果显示,HF2、HF3和BZ1的反应原性和免疫原性均优于HF1、XC1、BB1。HF2、HF3、BZ1在第10代均出现致病力减弱,其中BZ1传至20代、30代时仍呈现下降趋势,而HF2和HF3则趋于稳定。灭活全菌体HF2、HF3的血清抗体效价均由第10代1∶51 200下降至第30代的1∶12 800,而BZ1则由1∶12 800下降至1∶6 400。HF2、HF3、BZ1对昆明鼠和斑马鱼的免疫保护率分别为100%~80%、80%~60%、80%~40%和66.7%~60%、80%~60%、60%~53.3%。结果表明,HF2和HF3具备毒力强、抗原性好、遗传稳定的特性,可作为SS2制苗用菌株。

关键词: 猪链球菌2型, 疫苗菌株, 致病性, 抗原性, 稳定性

Abstract: To screen out the strains of Streptococcus suis type 2 (SS2) vaccine with strong pathogenicity, excellent antigenicity and relatively pure and stable genetic traits. In this study, 6 clinical isolates of SS2 virulent strains, numbered HF1, XC1, HF2, HF3, BZ1, BB1, were used as tested strains. The half lethal dose (LD₅₀) of strain was determined by modified Karber's method, the reactivity was determined by ELISA in detection of antibody titer of New Zealand white rabbit and Kunming mice serum, the immunogenicity was determined by immune challenge test in Kunming mice and zebrafish. At the same time, the tested strains were cultured continuously, the genetic stability of the tenth, twentieth, thirtieth generation of the tested strains were evaluated by the half lethal dose (LD₅₀), serum antibody titer and immune protection rate. The results showed that the LD₅₀ of BB1, BZ1, XC1, HF1, HF2 and HF3 for Kunming mice were 3.72×10⁹, 3.31×10⁸, 1.58×10⁹, 1.00×10⁹, 5.01×10⁸ and 3.24×10⁸ CFU·mL^-1, respectively, the LD₅₀ for zebrafish were 3.31×10³, 0.93×10², 6.03×10³, 3.39×10⁴, 0.62×10² and 2.34×10³ CFU·mL^-1, respectively. ELISA antibody titer determination, immune challenge test showed that the reactivity and immunogenicity of HF2, HF3 and BZ1 were better than those of HF1, XC1 and BB1. The pathogenicity of HF2, HF3, BZ1 decreased at the 10th generation, among which BZ1 still showed a downward trend at the 20th and 30th generations, while HF2 and HF3 tended to be stable. The serum antibody titers of inactivated whole cell HF3 and HF2 decreased from 1∶51 200 at the 10th generation to 1∶12 800 at the 30th generation, while BZ1 decreased from 1∶12 800 to 1∶6 400. The immune protection rates of HF2, HF3 and BZ1 were 100%-80%, 80%-60%, 80%-40% and 66.7%-60%, 80%-60%, 60%-53.3%, respectively. The results showed that HF2 and HF3 had the characteristics of strong virulence, excellent antigenicity and genetic stability which could be used as strains for SS2 seedling production.

Key words: Streptococcus suis type 2, vaccine strain, pathogenicity, antigenicity, stability

中图分类号:

S828

陈章, 吴华健, 毛天骄, 韩业芹, 孙裴, 魏建忠, 李东风, 李郁. 猪链球菌2型制苗用菌株的筛选[J]. 浙江农业学报, 2020, 32(1): 57-64.

CHEN Zhang, WU Huajian, MAO Tianjiao, HAN Yeqin, SUN Pei, WEI Jianzhong, LI Dongfeng, LI Yu. Screening of strain for inactivated vaccine of Streptococcus suis type 2[J]. , 2020, 32(1): 57-64.

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猪链球菌2型制苗用菌株的筛选

Screening of strain for inactivated vaccine of Streptococcus suis type 2

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