水貂巴氏杆菌病二价灭活疫苗的研究及其免疫效果的评价

发布时间：2018-02-26 11:28

本文关键词： 水貂巴氏杆菌病多杀性巴氏杆菌二价灭活疫苗免疫效果　出处：《吉林大学》2015年硕士论文　论文类型：学位论文

【摘要】：水貂巴氏杆菌病（Mink pasteurellosis）是由多杀性巴氏杆菌（Pasteurellamultocida）引发的水貂的一种急性、热性、败血性传染病，以出血性败血症及传染性肺炎为主要特征。本病常发生于春、秋两季，多呈散发，有时呈地方流行性，热带区域比温带区域发病率高，幼龄水貂感受性最强，发病率与死亡率均较高，该病的发生与流行给我国的水貂养殖业造成了严重的经济损失，因此在我国某些水貂养殖密集性地区进行水貂巴氏杆菌病病原的流行血清型研究，对于水貂巴氏杆菌病的防治有重要意义。目前，我国尚未有国家认证的预防水貂巴氏杆菌病的疫苗，，且多杀性巴氏杆菌血清型众多以及其耐药性的增强，这给水貂巴氏杆菌病的预防和治疗带来了诸多困难。为了有效地预防我国的水貂巴氏杆菌病，本研究采集了疑似水貂巴氏杆菌病病料进行病原的分离与鉴定，并进行了血清型分型、药物敏感性试验，选择致病性强、优势流行血清型菌株研制出水貂巴氏杆菌病二价灭活疫苗，并对其免疫效果进行了评价。首先，对从山东地区采集的2012~2013年疑似水貂巴氏杆菌病病料进行了病原的分离与鉴定。结果显示，从采集的53份疑似水貂巴氏杆菌病病料中检出多杀性巴氏杆菌28株，占总病例的52.8%。对分离菌株进行血清型分型，结果表明血清A型多杀性巴氏杆菌是主要流行型，占67.9%；血清B型是次要流行型，占32.1%。此外还对分离菌株进行了抗生素药物敏感性试验，结果表明大多数的分离菌株对多西环素、丁胺卡那霉素高度敏感。其次，通过对分离菌株致病力的测定，从分离菌株中选出连续传代对多杀性巴氏杆菌的菌体含量、致病力以及免疫原性均保持稳定的菌株血清A型WF403、血清B型WF405，将其4~6代菌株作为疫苗株来制备用于预防水貂巴氏杆菌病的二价灭活疫苗；对疫苗菌株培养条件如温度、培养时间等进行优化，制备了二批次的水貂巴氏杆菌病二价灭活疫苗：2014615、2014624；经物理性状检验、无菌检验、硫柳汞残留量及甲醛残留量的检验、超剂量安全性检验后，二批次的水貂巴氏杆菌病二价灭活疫苗的质量均合格，可以继续对灭活疫苗的免疫效果进行评价。最后，对研制的水貂巴氏杆菌病二价灭活疫苗进行了最小免疫剂量的测定以及免疫效果的评价。研究结果显示，该疫苗对水貂的最小免疫剂量是0.25mL，其常规免疫剂量是1.0mL；水貂在接种疫苗后14~21d，机体内的抗体水平达到高峰；测定水貂巴氏杆菌病二价灭活疫苗的免疫保护期至少是120d；该疫苗保存在2~8℃的条件下在9个月内对其免疫效果无显著的影响。疫苗进行临床试验的结果显示，进行免疫的水貂发病率为0.041%，显著低于未免疫时的发病率，说明此疫苗用于预防我国的水貂巴氏杆菌病具有良好的效果。本研究针对我国多杀性巴氏杆菌的优势流行血清型，研究制备了用于预防水貂巴氏杆菌病的二价灭活疫苗，为我国预防水貂巴氏杆菌病的发生奠定了理论依据和实践基础。
[Abstract]:Mink Pasteurellosis (Mink Pasteurellosis) by Pasteurella multocida (Pasteurellamultocida) is an acute, heat caused by mink, septic infectious disease, hemorrhagic septicemia and infectious pneumonia as the main feature. The disease often occurred in the spring and autumn season two, more sporadic, sometimes is endemic that area is higher than the temperate areas of tropical disease, the young mink feel the strongest, the incidence and mortality are high, causing serious economic losses to the occurrence and epidemic of the disease in our country mink industry, so the study of popular serotypes of Pasteurella multocida from mink mink breeding in our country in some water intensive areas that is important for the prevention of Pasteurellosis in mink. At present, vaccines for prevention of Pasteurellosis in mink has yet to have a national certification in China, and Pasteurella multocida serotype and drug resistance by many Strong, it brings many difficulties to the prevention and treatment of Pasteurellosis in mink mink pasteurellosis. In order to effectively prevent our country, this study collected the suspected Pasteurellosis in mink disease material for isolation and identification of pathogens, and the serotype, drug sensitive test, selection of high pathogenicity. The predominant serotypes, development of water mink Pasteurellosis two valent inactivated vaccine, and the immune effect was evaluated.
First of all, the collected from Shandong area 2012~2013 suspected Pasteurellosis in mink disease material for the isolation and identification of pathogen. The results showed that collected from 53 suspected Pasteurellosis in mink disease materials were detected in 28 strains of Pasteurella multocida, the total cases of 52.8%. of isolated strains were serotype. The results showed that the serum A of Pasteurella multocida is a major epidemic type, accounting for 67.9%; serum type B is a secondary popular type, accounting for 32.1%. in addition to the strains of antibiotic sensitivity test results show that most of the isolates of doxycycline, amikacin is highly sensitive.
Secondly, through the determination of the pathogenicity of isolates, selected continuous passage on cell content of Pasteurella multocida isolated from strain, pathogenicity and immunogenicity of serotype A WF403 strains were kept stable, serum B WF405, the 4~6 strain as vaccine strains to prepare for the prevention of mink PAP coli two bivalent inactivated vaccine; vaccine strains of culture conditions such as temperature, incubation time to optimize the preparation of the two batches of two mink Pasteurellosis bivalent inactivated vaccine: 20146152014624; the physical properties test, sterility test, thimerosal residues and formaldehyde residues, ultra safe dose after the test, the quality of two batches of two mink Pasteurellosis bivalent inactivated vaccine are qualified, can continue to evaluate the immune effect of inactivated vaccine.
Finally, the evaluation of the development of mink Pasteurellosis bivalent inactivated vaccine were two minimum immune dose determination and immune effect. The results showed that the minimum immune dose of the vaccine on mink is 0.25mL, the routine immunization dose is 1.0mL; mink after vaccination 14~ 21d, the antibody level reached a peak within the body immune protection; determination of mink Pasteurellosis bivalent inactivated vaccine at least two is 120d; the vaccine stored at 2~8 Deg. C within 9 months of its immune effect markedly. Vaccine clinical trial results showed that in the immune mink incidence rate was 0.041%, significantly lower than the onset when the immune rate, indicating that this vaccine for mink Pasteurellosis prevention in our country has a good effect.
Based on the dominant serotype of Pasteurella multocida in China, we studied the preparation of a two valent inactivated vaccine against Pasteurella multocida, which laid a theoretical foundation and practical basis for the prevention of Pasteurella multocida in China.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.92

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