【摘要】：馬腺疫是一種由C型鏈球菌馬鏈球菌馬亞種(Streptococcus equi subspecies equi,S.equi)引起的急性接觸性馬屬動(dòng)物傳染病,在我國牧區(qū)發(fā)病率很高,以2歲以下幼駒易感,給牧民造成巨大的經(jīng)濟(jì)損失,但尚無有效的預(yù)防和控制措施。為盡快緩解馬腺疫在我國牧區(qū)的流行情況,本項(xiàng)研究進(jìn)行了馬腺疫滅活疫苗制備方法的探討,試驗(yàn)獲得的病原菌為分離自馬腺疫重癥馬的野毒株(HLJ2011),利用該菌株制備滅活疫苗能夠?yàn)樾∈筇峁┝己玫谋Ｗo(hù)力。本文從HLJ2011菌株生物學(xué)特性及馬腺疫滅活疫苗小鼠免疫保護(hù)效力兩個(gè)方面進(jìn)行了系統(tǒng)的研究。結(jié)果如下:1.HLJ2011株生物學(xué)特性研究:本試驗(yàn)從病畜下頜破潰膿汁病料中分離出1株主要致病菌,對該菌的16S rRNA鑒定、生化鑒定、不同接種濃度生長曲線、菌落形成單位(Colony-Forming Unit,CFU)與OD值關(guān)系、馬腺疫鏈球菌毒力測定進(jìn)行了研究。結(jié)果表明,通過16S rRNA鑒定及生化鑒定該菌為S.equi,該菌不同接種濃度生長曲線表明,各濃度生長曲線基本相同,接種濃度越高,對數(shù)生長期越短;1%接種濃度培養(yǎng)8~10 h時(shí)CFU最大,可達(dá)1×109 CFU;用1×107 CFU腹腔攻毒8周齡Balb/c小鼠,死亡率100%,用改良寇氏法計(jì)算得出,該菌對小鼠半數(shù)致死量為5×105 CFU。2.馬腺疫滅活疫苗對小鼠免疫保護(hù)效力的研究方面:通過不同溫度和甲醛濃度完成HLJ2011滅活條件的篩選以及馬腺疫滅活疫苗對小鼠的免疫保護(hù)效力的研究。在4℃、28℃及37℃條件下不同甲醛濃度對滅活效果的影響試驗(yàn)表明,溫度越高或甲醛濃度越高,滅活時(shí)間越短。采用28℃、1%甲醛滅活12 h后,加入某高分子聚合物佐劑,經(jīng)無菌檢驗(yàn)后制成馬腺疫滅活疫苗,對5組小鼠進(jìn)行不同劑量的免疫后,分別進(jìn)行不同劑量攻毒試驗(yàn),結(jié)果表明小鼠經(jīng)106 CFU滅活疫苗免疫后,保護(hù)率為80%。本試驗(yàn)完成了對分離菌株HLJ2011的生物學(xué)特征研究,以及利用該菌株制備滅活疫苗對感染小鼠保護(hù)性試驗(yàn)。本項(xiàng)研究為我國馬腺疫病原學(xué)研究和疫苗研究奠定了重要基礎(chǔ)。
[Abstract]:Adenosis is an acute contagious disease of the genus Equine caused by (Streptococcus equi subspecies equi,S.equi (Streptococcus C Maya). The incidence of the disease is very high in pastoral areas of China. It is susceptible to colt under 2 years of age and causes great economic losses to herdsmen. However, there are no effective prevention and control measures. In order to alleviate the epidemic situation of horse adenosis in pastoral areas of China as soon as possible, the preparation method of inactivated vaccine against equine adenosis was studied in this study. The pathogen of the experiment was the wild virus strain (HLJ2011) isolated from the seriously infected horse. The inactivated vaccine prepared by this strain could provide good protection for mice. In this paper, the biological characteristics of HLJ2011 strain and the immune protective effect of inactivated equine adenosis vaccine in mice were studied systematically. The results were as follows: the biological characteristics of 1.HLJ2011 strain: a main pathogenic bacteria was isolated from the material of the disease animal's mandibular burst and pus. The 16s rRNA, biochemical identification and growth curve of different inoculation concentration were obtained. The relationship between colony forming unit (Colony-Forming Unit,CFU) and OD was studied. The results showed that the strain was S. equii by 16s rRNA identification and biochemical identification. The growth curve of different inoculation concentration was basically the same, the higher the inoculation concentration, the shorter the logarithmic growth period, and the maximum CFU was when 1% inoculation concentration was cultured for 810 h. 1 脳 10 ~ 9 CFU; could be used to attack 8 week-old Balb/c mice with 1 脳 10 ~ 7 CFU intraperitoneally. The mortality rate of Balb/c mice was 100%. Calculated by modified Kou's method, the median lethal dose of this strain to mice was 5 脳 10 ~ 5 CFU.2.. The study on the immune protection efficacy of inactivated vaccine against equine adenosis: the selection of HLJ2011 inactivation conditions was completed by different temperature and formaldehyde concentration, and the immune protection effect of inactivated vaccine against equine adenosis was studied. The effects of different formaldehyde concentrations on the inactivation effect at 4 鈩，

本文編號(hào)：2285513