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摘要:水体中抗生素与蓝藻共存可能性大，蓝藻在生理生态学方面受抗生素影响的研究却很少被关注。为此将研究四环素类抗生素中的金霉素对铜绿微囊藻合成及释放微囊藻毒素的急性影响作为本次实验的主题，用间接酶联免疫法对其微囊藻毒素含量进行检测。结果表明金霉素在一定的浓度范围内（1ppm、2ppm、5ppm）会促进藻细胞合成产生微囊藻毒素，金霉素浓度为5ppm时，藻细胞合成产生的微囊藻毒素最高为3161.066ng/L，其余试样均低于该浓度；超过该浓度范围（10ppm、20ppm）可能会抑制藻细胞合成产生微囊藻毒素，但释放到环境中的微囊藻毒素的含量则会随着金霉素浓度的增加而升高，金霉素浓度为20ppm时藻细胞释放到环境中的微囊藻毒素含量最高为952.972ng/L。通过对本实验的研究，我们可以了解抗生素对蓝藻释放微囊藻毒素的影响，对合理使用抗生素有一定的指导意义。
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关键词：铜绿微囊藻，金霉素，微囊藻毒素，间接酶联免疫法4167
Aquatic toxicity assessment of Chlortetracycline hydrochloride
Abstract: It is possible that antibiotics could be co-existed with cyanobacteria in water. However, research about the effects of antibiotics on Microcystis aeruginosa gained little attention. Therefore, investigating the acute effects of chlortetracycline, one of the tetracycline antibiotic, on MC-LR release in Microcystis aeruginosa were carried out in the present study. The indirect enzyme-linked immunosorbent assay (ELISA) is applied to monitor the content of MC-LR. The results showed that in a certain concentration range (1 ppm, 2 ppm, 5 ppm), chlortetracycline could induce Microcystis aeruginosa to synthesize and production of MC-LR. At the concentration of 5 ppm, Microcystis aeruginosa synthesized and produced the highest content of MC-LR which is 3161.066 ng/L. The other samples are less. During the concentrations of 10 ppm and 20 ppm, chlortetracycline may inhibit Microcystis aeruginosa synthesis and production of MC-LR, and with the increase of the concentration of chlortetracycline, MC-LR content released to the environment will be increased. At the concentration of 20 ppm, content of MC-LR released into the environment is up to 952.972 ng/L. Through the present study, the effects of antibiotics on MC-LR release in Microcystis aeruginosa could be understood, and it has a significant meaning to use antibiotics rationally.
Keywords: cyanobacteria; Chlortetracycline hydrochloride; MC-LR; enzyme-linked immunosorbent assay (ELISA)
目录
1 前言 1
1.1 选题背景 1
1.1.1 抗生素的基本状况 1
1.1.2 金霉素的物理化学性质 2
1.1.3 微囊藻毒素(Microcystins，MCYSTs，MCs)的物理化学性质 2
1.1.4 微囊藻毒素的研究进展 3
2 研究方案与方法 5
2.1 实验铜绿微囊藻的培养 5
2.2 实验方案 6
3 实验部分 7
3.1 实验材料与试剂 7
3.2 金霉素储备液配制 7
3.3 铜绿微囊藻细胞密度与吸光度关系测定 7
3.4 实验藻样的接种与处理 8
3.5 染毒培养48h后接种藻液吸光度及藻细胞浓度 8
3.6 染毒48h后藻液细胞外微囊藻毒素的提取 8
3.7 染毒后48h藻液细胞内和细胞外总微囊藻毒素提取 9
3.8 微囊藻毒素（MC-LR）含量的测定 9
3.8.1 间接酶联免疫法（Indirect ELISA）原理 10
3.8.2 间接酶联免疫法（Indirect ELISA）实验过程 10
3.9 实验数据分析 11