【正文】 ...................................................... 9 實(shí)驗(yàn)儀器 ............................................................................................................... 9 實(shí)驗(yàn)材料 ............................................................................................................... 9 實(shí)驗(yàn)藥品 ............................................................................................................. 11 預(yù)實(shí)驗(yàn) ........................................................................................................................... 11 BG11 培養(yǎng)液的配置 ............................................................................................ 11 高壓蒸氣滅菌 ..................................................................................................... 12 銅綠微囊藻的 接種 ............................................................................................. 12 銅綠微囊藻的 培養(yǎng) ............................................................................................. 12 實(shí)驗(yàn)方案論證 ............................................................................................................... 12 3. 實(shí)驗(yàn)方案與方法 .................................................................................................................... 14 四環(huán)素儲(chǔ)備液的配置 ................................................................................................... 14 銅綠微囊藻細(xì)胞數(shù)與吸光度的關(guān)系 ........................................................................... 14 實(shí)驗(yàn)原理 ............................................................................................................. 14 實(shí)驗(yàn)方法 ............................................................................................................. 15 銅綠微囊藻生長(zhǎng)曲線及抑制率 ................................................................................... 15 銅綠微囊藻 pH 變化規(guī)律 ............................................................................................ 16 考馬斯亮藍(lán)染色法測(cè)定蛋白質(zhì)含量 ........................................................................... 16 實(shí)驗(yàn)原理 ............................................................................................................. 16 試劑配制 ............................................................................................................. 16 實(shí)驗(yàn)方法 ............................................................................................................. 16 高效液相色譜檢測(cè)四環(huán)素在銅綠 微囊藻液中的降解 ............................................... 17 實(shí)驗(yàn)原理 ............................................................................................................. 17 實(shí)驗(yàn)特點(diǎn) ............................................................................................................. 17 實(shí)驗(yàn)方法 ............................................................................................................. 18 4. 結(jié)果與討論 ............................................................................................................................ 20 銅綠微囊藻細(xì)胞數(shù)與吸光度的關(guān)系 ........................................................................... 20 生長(zhǎng)曲線與抑制率 ....................................................................................................... 20 銅綠微囊藻 pH 變化規(guī)律 ............................................................................................ 25 考馬斯亮藍(lán)染色法測(cè)定蛋白質(zhì)含量 ........................................................................... 26 高效液相色譜檢測(cè)四環(huán)素在銅綠微囊藻液中的降解 ............................................... 28 標(biāo)準(zhǔn)曲線 ............................................................................................................. 28 降解曲線 ............................................................................................................. 31 5. 結(jié)論 ........................................................................................................................................ 37 四環(huán)素對(duì)藍(lán)藻的急性毒性研究 1 1. 引言（緒論） 抗生素的污染現(xiàn)狀 抗生素是一類生物（主要是真菌、放線菌或細(xì)菌等微生物）在其代謝過程中產(chǎn)生的具有殺滅或抑制他種生物（主要是微生物）作用的化學(xué)物質(zhì)。 Acute toxicity。 關(guān)鍵詞： 四環(huán)素、銅綠微囊藻、急性毒性、抑制、降解 Acute toxicity of tetracycline on Microcystis aeruginosa Abstract: Tetracycline antibiotics are widely used in the world. This is one of the largest types of antibiotics, which can remain in the environment and bring potential environmental risks. In this study, the ecotoxicity of tetracyclines on cyanobacteria Microcystis aeruginosa was studied. Growth inhibition of this kind of algae exposure to tetracycline was measured. pH values, protein content, were also studied to determine the acute toxicity of tetracycline on M. aeruginosa. The results showed that at low concentration 1 mg/L tetracycline can stimulate the growth of M. aeruginosa, and in prolonged exposure to high concentrations of tetracycline, M. aeruginosa growth was suppressed. This paper also detected the degradation of tetracycline in M. aeruginosa culture medium by high performance liquid chromatography (HPLC). The results showed that the reaction is a firstorder kiics reaction, Ln (tetracycline concentration) and the reaction time showed linear relationship. And discussed the future research directions, aimed to provide valuable reference for the ecological risk assessment of the antibiotics. KeyWords: Tetracycline。 本文同時(shí)通過高效液相色譜法檢測(cè)了四環(huán)素在銅綠微囊藻 液中的降解情況，結(jié)果表明： 該反應(yīng)屬于一級(jí) 動(dòng)力學(xué) 反應(yīng)， Ln（四環(huán)素濃度）與反應(yīng)時(shí)間呈線性關(guān)系。 并以水華藍(lán)藻銅綠微囊藻為研究對(duì)象，通過四環(huán)素對(duì)銅綠微囊藻的生長(zhǎng)抑制、 四環(huán)素 對(duì)藻液pH 值的影響、 四環(huán)素 對(duì)蛋白質(zhì)含量的影響，研究四環(huán)素對(duì)藍(lán)藻的急性毒性。 指導(dǎo)老師： 葉璟 審核意見： 教研室主任： 曹萍 四環(huán)素對(duì)藍(lán)藻的急性毒性研究 摘要 ： 四環(huán)素 類抗生素 是 目前 使用最廣泛、用量最大的抗生素種類之一 , 其在環(huán)境中的大量殘留帶來了潛在的環(huán)境風(fēng)險(xiǎn)。 5～ 14 周：實(shí)驗(yàn)室實(shí)驗(yàn)。 Smyth, M. R.。 Carmichael, W. W., Analysis and purification of toxic peptides from cyanobacteria by reversedphase highperformance liquid chromatography. Journal of Chromatography A 1988,