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青島農(nóng)業(yè)大學(xué) 畢 業(yè) 論 文 題 目: 反芻動(dòng)物源性枯草芽孢桿菌的分離與鑒定 姓 名: 學(xué) 院: 動(dòng)物科技學(xué)院 專 業(yè): 動(dòng)物醫(yī)學(xué) 班 級(jí): 1001 學(xué) 號(hào): 20xx5254 指導(dǎo)教師: 溫建新 20xx 年 6 月 5 日 1 反芻動(dòng)物 源性 枯草芽孢桿菌 的分離與鑒定 動(dòng)物醫(yī)學(xué)專業(yè) 袁如意 指導(dǎo)教師 溫建新 摘要: 從健康 荷斯坦成年 奶 牛 新鮮 糞便中分離純化出 1 株 疑似 芽孢桿菌 屬菌株 ,編號(hào)為 K。對(duì)該菌 株 進(jìn)行形態(tài)學(xué)觀察 、生化試驗(yàn)、藥敏試驗(yàn)及安全性試驗(yàn)等 。 試驗(yàn)結(jié)果表明,該菌 菌體 革蘭氏染色陽(yáng)性,芽孢染色為綠色, 呈長(zhǎng)桿狀,芽孢中生,橢圓 ;能發(fā)酵 糖醇,吲哚試驗(yàn)陽(yáng)性,觸媒試驗(yàn)陽(yáng)性,能產(chǎn)生淀粉酶,并且可以液化明膠 ,生化試驗(yàn)結(jié)果表明 K 菌為枯草芽孢桿菌;該菌對(duì) 鏈霉素、青霉素、諾氟沙星、四環(huán)素耐藥性強(qiáng) ,而對(duì)慶大霉素、卡那霉素、環(huán)丙沙星等 抗生素表現(xiàn)敏感 ; 動(dòng)物 安全 試驗(yàn)表明,灌服 濃度為 1011cfu/mL 菌液的小鼠精神狀態(tài)良好,無(wú) 死亡現(xiàn)象。 本試驗(yàn)為 獲得反芻動(dòng)物的有益菌種 及 研制反芻動(dòng)物微生態(tài)制劑 奠定基礎(chǔ)。 關(guān)鍵詞: 反芻動(dòng)物 ;枯草芽孢桿菌; 分離 ; 鑒定 2 Isolation and Identification of Bacillus subtilis Strains from Ruminants Student majoring in veterinary medicine Yuan Ruyi Tutor Wen Jianxin Abstract: One bacillus strain was isolated and purified from the faeces of healthy Holstein named K. Morphological observation, biochemical tests, chemosensitivity test and security experiment were conducted. Experimental results showed that strain K Gram stain was positive, spore staining was green. Its shape was a long rodshaped bacteria, and spores was oval . Common sugar alcohols could be fermented。 Indole test was positive。 Catalyst test was positive。 It could produce amylase。 It also could liquefy gelatin. Susceptibility testing showed that the bacteria strongly resisted to streptomycin, penicillin, norfloxacin,and tetracycline and that it was sensitive to gentamicin, kanamycin and ciprofloxacin. Animal safety trials showed that Mice fed bacterias 1011cfu/mL w ere in good condition and no mortality. therefore, the K was identified as Bacillus subtilis strain. So it provided theoretical basis for developing probiotic preparations. Key words: ruminants。 Bacillus Subtilis。 isolation。 identification 3 引言 枯草芽孢桿菌是存在于動(dòng)物腸道中的非致病需氧菌 [1],可降低腸道內(nèi)氧濃度,產(chǎn)生的枯草菌素、多粘菌素、短桿菌肽等活性物質(zhì)能顯著降低腸道