基于氧微电极的生物膜内反应动力学参数的原位测定研究
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摘要
在生物膜模型的应用与研究过程中,如何准确地获得生物膜的反应动力学参数和传质动力学参数是至关重要的,其准确性在很大程度上决定了生物膜模型的预测能力。由于生物膜内部结构和反应过程很复杂,要准确获得生物膜动力学参数比较困难。本文提出了两种获得生物膜动力学参数的原位测定方法:“扩散-反应模型法”和“呼吸速率测量法”。在完成分离式氧微电极结构优化并构建起生物膜动力学参数原位测定系统的基础上,用“扩散-反应模型法”和“呼吸速率测量法”对生物膜动力学参数进行原位测定研究。
“扩散-反应模型法”是在生物膜处于内源呼吸阶段及生长阶段用氧微电极检测生物膜内的溶解氧分布曲线,并结合稳态条件下的扩散-反应方程对生物膜动力学参数进行估值。用该方法检测得到的生物膜内源呼吸速率、衰减系数及氧半饱和常数分别为8.52mgO_2/(gVSS·h)、0.0035h~(-1)、1.00mg/L。与活性污泥系统相比,用该方法获得的内源呼吸速率和衰减系数较小,对氧的半饱和常数较大。
“呼吸速率测量法”是在没有破坏生物膜结构的基础上,用微型加样管在生物膜内某一深度注射微量基质溶液,同时氧微电极在该加样点检测微生物的呼吸速率曲线,并结合Monod方程计算该点的动力学参数。在得到生物膜内不同深度微观点的动力学参数后用算术平均值法对生物膜表观动力学参数进行估值。用该方法获得的生物膜产率系数、最大比增长速率和基质半饱和常数的平均值分别为0.44、4.00d~(-1)、41.06mg/L。与破坏生物膜结构后用呼吸速率测量法检测的结果相比,原位测定方法获得的生物膜产率系数与之相近,最大比增长速率较小,对基质的半饱和常数较大。同时研究结果表明生物膜内各深度微生物的产率系数变化不大,在0.40-0.45范围内波动;最大比增长速率沿着生物膜深度方向逐渐降低,从表层的5.74 d~(-1)降到深层的2.45 d~(-1);对基质的半饱和常数沿生物膜深度方向逐渐增大,从表层的36.63mg/L升高到深层的48.04mg/L。
The methods to obtain the kinetic parameters of biofilm accurately are very important when the biofilm models are used in research, and the forecasting ability of the biofilm models depends on the accuracy of the kinetic parameters. As the complexity of the inner structure and the reaction processes of biofilm, it is very difficult to obtain the kinetic parameters of biofilm exactly. In this study, two kinds of in situ methods to detect the biofilm kinetic parameters were established: the“diffusion-reaction model mothod”and the“oxygen uptake rate method”. After optimizing the configuration of separated oxygen microelectrode and establishing the in-situ detecting system, the two in-situ methods were used to detect the biofilm kinetic parameters.
The“diffusion-reaction model method”could be used to estimate the biofilm kinetic parameters by detecting the oxygen distribution profiles in the biofilm in the endogenous phase and growing phase with an oxygen microelectrode. After obtaining the oxygen distribution profiles, the biofilm kinetic parameters could be calculated based on the diffusion-reaction equations under the stabilized conditions. The average values of the OURin (endogenous respiratory rate), Kd (endogenous decay rate) and the Ko (Monod half saturation coefficient to the oxygen) in the biofilm which were estimated by this in-situ method were 8.52 mg O2/ (gVSS·h), 0.0035h-1 and 1.0mg/L respectively. Compared with the activated sludge system, the endogenous respiratory rate and the endogenous decay rate were smaller; the Monod half saturation coefficient was higher.
The“oxygen uptake rate method”could be used to estimate the biofilm kinetic parameters by combining the Monod equation and the respiration rate profiles which were detected with an oxygen microelectrode after injecting minim of substrate solution to some depth of the biofilm. After acquiring the kinetic parameters of the microcosmic detected points in the different depths of the biofilm, the apparent values of the biofilm kinetic parameters could be attained by averaging the microcosmic parameters. The apparent values of the Y (yield coefficient),μmax(maximum growing rate)and the Ks (Monod half saturation coefficient to the glucose) were 0.44, 4.00d-1, 41.06mg/L respectively by the in situ method of“oxygen uptake rate”. Compared with the values after destroying the biofilm structure, the Y,μmaxand the Ks which were detected using the in situ method were approximate, smaller and higher respectively. Further more, the in situ method of“oxygen uptake rate”could study the kinetic parameters of the microorganisms in the different depths of the biofilm. The results showed that the Y in the different depths of the biofilm which varied from 0.40 to 0.45 didn’t have much diversification; theμmaxdecreased gradually with the depth increasing, which changed from 5.74 d-1 on the surface of biofilm to 2.45 d-1 at the bottom of biofilm; the Ks increased gradually with the depth increasing, which changed from 36.63mg/L on the surface of biofilm to 48.04mg/L at the bottom of biofilm.
“扩散-反应模型法”是在生物膜处于内源呼吸阶段及生长阶段用氧微电极检测生物膜内的溶解氧分布曲线,并结合稳态条件下的扩散-反应方程对生物膜动力学参数进行估值。用该方法检测得到的生物膜内源呼吸速率、衰减系数及氧半饱和常数分别为8.52mgO_2/(gVSS·h)、0.0035h~(-1)、1.00mg/L。与活性污泥系统相比,用该方法获得的内源呼吸速率和衰减系数较小,对氧的半饱和常数较大。
“呼吸速率测量法”是在没有破坏生物膜结构的基础上,用微型加样管在生物膜内某一深度注射微量基质溶液,同时氧微电极在该加样点检测微生物的呼吸速率曲线,并结合Monod方程计算该点的动力学参数。在得到生物膜内不同深度微观点的动力学参数后用算术平均值法对生物膜表观动力学参数进行估值。用该方法获得的生物膜产率系数、最大比增长速率和基质半饱和常数的平均值分别为0.44、4.00d~(-1)、41.06mg/L。与破坏生物膜结构后用呼吸速率测量法检测的结果相比,原位测定方法获得的生物膜产率系数与之相近,最大比增长速率较小,对基质的半饱和常数较大。同时研究结果表明生物膜内各深度微生物的产率系数变化不大,在0.40-0.45范围内波动;最大比增长速率沿着生物膜深度方向逐渐降低,从表层的5.74 d~(-1)降到深层的2.45 d~(-1);对基质的半饱和常数沿生物膜深度方向逐渐增大,从表层的36.63mg/L升高到深层的48.04mg/L。
The methods to obtain the kinetic parameters of biofilm accurately are very important when the biofilm models are used in research, and the forecasting ability of the biofilm models depends on the accuracy of the kinetic parameters. As the complexity of the inner structure and the reaction processes of biofilm, it is very difficult to obtain the kinetic parameters of biofilm exactly. In this study, two kinds of in situ methods to detect the biofilm kinetic parameters were established: the“diffusion-reaction model mothod”and the“oxygen uptake rate method”. After optimizing the configuration of separated oxygen microelectrode and establishing the in-situ detecting system, the two in-situ methods were used to detect the biofilm kinetic parameters.
The“diffusion-reaction model method”could be used to estimate the biofilm kinetic parameters by detecting the oxygen distribution profiles in the biofilm in the endogenous phase and growing phase with an oxygen microelectrode. After obtaining the oxygen distribution profiles, the biofilm kinetic parameters could be calculated based on the diffusion-reaction equations under the stabilized conditions. The average values of the OURin (endogenous respiratory rate), Kd (endogenous decay rate) and the Ko (Monod half saturation coefficient to the oxygen) in the biofilm which were estimated by this in-situ method were 8.52 mg O2/ (gVSS·h), 0.0035h-1 and 1.0mg/L respectively. Compared with the activated sludge system, the endogenous respiratory rate and the endogenous decay rate were smaller; the Monod half saturation coefficient was higher.
The“oxygen uptake rate method”could be used to estimate the biofilm kinetic parameters by combining the Monod equation and the respiration rate profiles which were detected with an oxygen microelectrode after injecting minim of substrate solution to some depth of the biofilm. After acquiring the kinetic parameters of the microcosmic detected points in the different depths of the biofilm, the apparent values of the biofilm kinetic parameters could be attained by averaging the microcosmic parameters. The apparent values of the Y (yield coefficient),μmax(maximum growing rate)and the Ks (Monod half saturation coefficient to the glucose) were 0.44, 4.00d-1, 41.06mg/L respectively by the in situ method of“oxygen uptake rate”. Compared with the values after destroying the biofilm structure, the Y,μmaxand the Ks which were detected using the in situ method were approximate, smaller and higher respectively. Further more, the in situ method of“oxygen uptake rate”could study the kinetic parameters of the microorganisms in the different depths of the biofilm. The results showed that the Y in the different depths of the biofilm which varied from 0.40 to 0.45 didn’t have much diversification; theμmaxdecreased gradually with the depth increasing, which changed from 5.74 d-1 on the surface of biofilm to 2.45 d-1 at the bottom of biofilm; the Ks increased gradually with the depth increasing, which changed from 36.63mg/L on the surface of biofilm to 48.04mg/L at the bottom of biofilm.
引文
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