本文關(guān)鍵詞： 水貂 肺炎病原菌 藥敏試驗(yàn) 綠膿桿菌 多價(jià)滅活疫苗　出處：《東北林業(yè)大學(xué)》2015年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：水貂細(xì)菌性肺炎是困擾水貂養(yǎng)殖業(yè)的難題,為了調(diào)查引起該病的主要病原菌并探索安全有效的防治方法,2011年至2013年,對(duì)從山東、遼寧兩省送檢的水貂肺炎病例進(jìn)行病原菌分離,共分離到38株細(xì)菌,結(jié)合生化鑒定和分子生物學(xué)鑒定方法,最終確定包括31株綠膿桿菌,4株肺炎克雷伯氏菌,1株臭鼻克雷伯氏菌,1株產(chǎn)酸克雷伯氏菌和1株與綠膿桿菌混合感染的鮑曼不動(dòng)桿菌。綠膿桿菌包括三個(gè)血清型G型(22/31)、C型(6/31)、B型(3/31),G型為主要流行血清型。藥敏試驗(yàn)結(jié)果顯示所分離到的38株肺炎病原菌均對(duì)亞胺培南和美洛培南敏感,對(duì)氨芐青霉素和阿莫西林／克拉維酸耐藥。所有分離到的病原菌均對(duì)小鼠有較強(qiáng)致病性。取綠膿桿菌G、C、B血清型各一株,做標(biāo)準(zhǔn)抗原,并免疫家兔,采集血清做標(biāo)準(zhǔn)抗體,通過(guò)方陣凝集試驗(yàn)確定標(biāo)準(zhǔn)抗原抗體最適反應(yīng)濃度。將分離到的綠膿桿菌菌株免疫小鼠,采集血清,利用標(biāo)準(zhǔn)抗原檢測(cè)免疫后小鼠血清效價(jià),最終篩選了PA-L14(G型)、PA-L05(B型)、PA-L06(C型)三株免疫原性較好的菌株作為疫苗候選菌株。以終濃度為0.3%甲醛,經(jīng)37℃、24h對(duì)菌液進(jìn)行滅活處理。疫苗包含9×109CFU/mL的G型和B型綠膿桿菌,6×109CFU/mL的C型綠膿桿菌,配以20%鋁膠佐劑,0.01%的硫柳汞,免疫健康無(wú)抗體的水貂進(jìn)行抗體效價(jià)跟蹤,結(jié)果顯示免疫水貂在半月內(nèi)機(jī)體可產(chǎn)生較高水平的抗體,該抗體水平可維持3個(gè)月,滿足了在綠膿桿菌出血性肺炎流行季節(jié)對(duì)水貂的保護(hù)要求,大規(guī)模動(dòng)物免疫回訪結(jié)果顯示,疫苗速發(fā)接種反應(yīng)發(fā)生率低,預(yù)后良好,對(duì)預(yù)防接種和發(fā)病后緊急接種,均具有較好的保護(hù)性。
[Abstract]:Mink bacterial pneumonia is a difficult problem for mink breeding. In order to investigate the main pathogens causing the disease and explore safe and effective control methods, from 2011 to 2013, from Shandong Province. A total of 38 strains of bacteria were isolated from mink pneumonia cases in Liaoning Province. 31 strains of Pseudomonas aeruginosa were identified by biochemical identification and molecular biological identification. Four strains of Klebsiella pneumoniae, one strain of Klebsiella sinensis and one strain of Acinetobacter baumannii co-infected with Pseudomonas aeruginosa. Pseudomonas aeruginosa consists of three serotypes G (22 / 31). C / C / 31 / B was the main epidemic serotype. The results of drug sensitivity test showed that all the 38 strains of pathogenic bacteria were sensitive to imipenem and meropenem. Ampicillin and amoxicillin / clavulanic acid were resistant to ampicillin and amoxicillin / clavulanic acid. All the isolated pathogens were highly pathogenic to mice. The collected serum was used as standard antibody and the optimum reaction concentration of standard antigen antibody was determined by square array agglutination test. The isolated strains of Pseudomonas aeruginosa were immunized with mice and serum was collected. The serum titer of immunized mice was detected by standard antigen, and the PA-L14(G type PA-L05B was screened out. PA-L06(C type) three strains with good immunogenicity were used as vaccine candidates. The final concentration of formaldehyde was 0.3% at 37 鈩，

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