本文關(guān)鍵詞： 水貂巴氏桿菌病 多殺性巴氏桿菌 二價滅活疫苗 免疫效果　出處：《吉林大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：水貂巴氏桿菌�。∕ink pasteurellosis）是由多殺性巴氏桿菌（Pasteurellamultocida）引發(fā)的水貂的一種急性、熱性、敗血性傳染病，以出血性敗血癥及傳染性肺炎為主要特征。本病常發(fā)生于春、秋兩季，多呈散發(fā)，有時呈地方流行性，熱帶區(qū)域比溫帶區(qū)域發(fā)病率高，幼齡水貂感受性最強(qiáng)，發(fā)病率與死亡率均較高，該病的發(fā)生與流行給我國的水貂養(yǎng)殖業(yè)造成了嚴(yán)重的經(jīng)濟(jì)損失，因此在我國某些水貂養(yǎng)殖密集性地區(qū)進(jìn)行水貂巴氏桿菌病病原的流行血清型研究，對于水貂巴氏桿菌病的防治有重要意義。目前，我國尚未有國家認(rèn)證的預(yù)防水貂巴氏桿菌病的疫苗，，且多殺性巴氏桿菌血清型眾多以及其耐藥性的增強(qiáng)，這給水貂巴氏桿菌病的預(yù)防和治療帶來了諸多困難。為了有效地預(yù)防我國的水貂巴氏桿菌病，本研究采集了疑似水貂巴氏桿菌病病料進(jìn)行病原的分離與鑒定，并進(jìn)行了血清型分型、藥物敏感性試驗(yàn)，選擇致病性強(qiáng)、優(yōu)勢流行血清型菌株研制出水貂巴氏桿菌病二價滅活疫苗，并對其免疫效果進(jìn)行了評價。 首先，對從山東地區(qū)采集的2012~2013年疑似水貂巴氏桿菌病病料進(jìn)行了病原的分離與鑒定。結(jié)果顯示，從采集的53份疑似水貂巴氏桿菌病病料中檢出多殺性巴氏桿菌28株，占總病例的52.8%。對分離菌株進(jìn)行血清型分型，結(jié)果表明血清A型多殺性巴氏桿菌是主要流行型，占67.9%；血清B型是次要流行型，占32.1%。此外還對分離菌株進(jìn)行了抗生素藥物敏感性試驗(yàn)，結(jié)果表明大多數(shù)的分離菌株對多西環(huán)素、丁胺卡那霉素高度敏感。 其次，通過對分離菌株致病力的測定，從分離菌株中選出連續(xù)傳代對多殺性巴氏桿菌的菌體含量、致病力以及免疫原性均保持穩(wěn)定的菌株血清A型WF403、血清B型WF405，將其4~6代菌株作為疫苗株來制備用于預(yù)防水貂巴氏桿菌病的二價滅活疫苗；對疫苗菌株培養(yǎng)條件如溫度、培養(yǎng)時間等進(jìn)行優(yōu)化，制備了二批次的水貂巴氏桿菌病二價滅活疫苗：2014615、2014624；經(jīng)物理性狀檢驗(yàn)、無菌檢驗(yàn)、硫柳汞殘留量及甲醛殘留量的檢驗(yàn)、超劑量安全性檢驗(yàn)后，二批次的水貂巴氏桿菌病二價滅活疫苗的質(zhì)量均合格，可以繼續(xù)對滅活疫苗的免疫效果進(jìn)行評價。 最后，對研制的水貂巴氏桿菌病二價滅活疫苗進(jìn)行了最小免疫劑量的測定以及免疫效果的評價。研究結(jié)果顯示，該疫苗對水貂的最小免疫劑量是0.25mL，其常規(guī)免疫劑量是1.0mL；水貂在接種疫苗后14~21d，機(jī)體內(nèi)的抗體水平達(dá)到高峰；測定水貂巴氏桿菌病二價滅活疫苗的免疫保護(hù)期至少是120d；該疫苗保存在2~8℃的條件下在9個月內(nèi)對其免疫效果無顯著的影響。疫苗進(jìn)行臨床試驗(yàn)的結(jié)果顯示，進(jìn)行免疫的水貂發(fā)病率為0.041%，顯著低于未免疫時的發(fā)病率，說明此疫苗用于預(yù)防我國的水貂巴氏桿菌病具有良好的效果。 本研究針對我國多殺性巴氏桿菌的優(yōu)勢流行血清型，研究制備了用于預(yù)防水貂巴氏桿菌病的二價滅活疫苗，為我國預(yù)防水貂巴氏桿菌病的發(fā)生奠定了理論依據(jù)和實(shí)踐基礎(chǔ)。
[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.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.92

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