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1、l 2013 年度本科生毕业论文 生化需氧量传感器生物膜电极的 初步研究 学 院 ： 理工学院 专 业： 环境工程 年 级： 2009 级 学生姓名： 学 号： 09222069 导师及职称： 讲师 2013 年 5 月 l Study on efficient microbial membrance with marine yeast applied in BOD sensor for fishery seawater off shore Department: School of Sicence and Technology Major: Environment Engineering G
2、rade: 2009 Students Name: Yongfu Liu Student No.: 09222069 Tutor: Lecturer Rui Hu Finished by May, 2013 l 摘 要 生化需氧量(BOD)是监测水体中可生物降解的有机物含量的一个重要参数, 是目前最常用的水体有机污染综合指标之一。其标准测定方法为五日法,以被检 验的水样在 201下,5 天内的耗氧量为代表,称其为 5 日生化需氧量,简称 BOD5。但该方法操作较复杂,耗时长,结果准确度及重现性差,无法满足当前环 境监测中快速测定的要求。而日益得到广泛应用的微生物传感器法能够快速、 准确测定水体
3、中 BOD,并可实现在线连续测定,更加适应于现代环境监测和管理 的要求。本文阐述了 BOD 微生物传感器技术的研究进展及应用。分析了微生物 传感器的工作原理及其在水质测定中的优点,重点研究了其核心部分微生物膜的 研究现状以及制备过程中存在的问题。针对微生物膜的研究现状,以菌种选育和 固定化方法的选择作为本次实验研究的重点。本文综述了微生物固定化方法在 各类废水处理中的研究现状,对固定化方法进行了分类,并对其性能作了比较。 通过比较三种微生物膜电极的底物适用范围、线性范围及其测量结果的准 度和精密度，选择了对数生长期的汉逊德巴利海洋酵母 Debaryomyces hansenii HXY-09
4、构建的微生物膜电极作为实验所用电极。进一步对近海养殖水体 BOD 含量的检测结果有显著影响的待测样品底液 pH、电极工作温度和底液离子浓度 3 个因素为自变量，以传感器的响应电流为响应值，研究了外部检测条件的变 化对实验结果的影响，为该复合功能菌群进一步研究近海养殖水体 BOD 传感器 奠定基础。 关键词：生化需氧量；微生物膜；微生物固定化；微生物传感器；溶氧电极 l ABSTRACT Biochemical oxygen demand (BOD) is an important parameter of the organic matter content of biodegradable
5、monitoring in water, is one of the comprehensive index of organic pollution in water is the most commonly used at present. The standard determination method for five days, to water samples tested at 20 1 , oxygen consumption of 5 days as the representative, called the 5 day biochemical oxygen demand
6、, referred to as BOD5. But the method is complicated, time- consuming, accuracy and reproducibility is poor, unable to meet the rapid determination of present environmental monitoring requirements. It has been widely used in microbial sensor method can fast, accurate determination of BOD in water, a
7、nd can realize continuous online measurement, more suitable for modern environmental monitoring and management requirements. In this paper, the research progress and application of BOD microbial sensor technology. Analysis of the principle of microbial sensor and its advantages in measurement in wat
8、er, has focused on its core part of biofilm research status and problems in the process of preparation. According to the research status of microbial membrane, to the breeding and the immobilized method chosen as the key research of this experiment. This paper summarized the present situation of res
9、earch on microbial immobilization method in all kinds of waste water disposal, the immobilization methods are classified, and compared its performance. The Debaryomyces hansenii based biosensor is characterized by the highest sensitivity and, accordingly, the minimal lower limit of linear range. Thu
10、s, the Debaryomyces hansenii biosensor has the best characteristics of those tested. Response Surface Methodology (RSM) was used to investigate the effects of pH, Temperature and concentration buffers on the results. A 33 factorial design was performed to optimize the conditions. The experimental re
11、sults showed that the optimal conditions are: pH of 7.7, temperature 20 and concentration buffers 40 mmol/L, and the small relative error of 1% between biosensor and BOD5. This study l indicates that Debaryomyces hansenii based biosensor can be used as a reliable method to determine BOD in seawater,
12、 the linear fluctuant coefficients (R2) in the range of 9.3422 mg/dm3 when a glucose/glutamate BOD standard was applied. Such device provides a low-cost, easy-operated, fast-response, sensitive and reliable method to measure BOD in seawater, and also is suitable for on-line measurements. Keywords: B
13、iochemical Oxygen Demand(BOD); Microbial Film; Immobilized Microorganism; Microbial Sensor; Dissolved Oxygen Eletrode l 目 录 第一章 绪论.1 1.1 问题的提出 .1 1.1.1 传统测量方法存在的问题 .1 1.1.2 BOD 微生物传感器的产生.1 1.2 课题研究的目的和意义.2 1.3 课题研究的内容.3 第二章 实验材料和方法.4 2.1 材料.4 2.1.1 菌种与来源 .4 2.1.2 培养基 .4 2.1.3 试剂和仪器 .4 2.2 海洋酵母活菌总数与生长周期10.4 2.3 BOD 标准溶液的配制 .5 2.4 微生物膜电极的制备.5 2.5 近海养殖水体中 BOD 微生物传感系统的组装.6 2.6 样品的测定.8 第三章 结论和分析.
