【作者】 胡子全；

【导师】 赵海泉；

【作者基本信息】 安徽农业大学 ， 微生物学， 2008， 硕士

【摘要】 本研究利用YG培养基从巢湖底泥、校园池塘底泥和南淝河底泥等不同地点采集的污泥中分离纯化了66株细菌,其中30株细菌生长旺盛,将这些菌株分别点种到限磷与过磷的葡萄糖-MOPS培养基上培养2d,在两种培养基上都产生蓝斑的菌株有11株,即为初选聚磷菌。将这11株细菌进行厌氧好氧连续培养,进行内聚物的染色,其中有7株细菌能合成内聚-β-羟基丁酸和多聚磷酸盐;好氧培养24时,在7株能合成内聚-β-羟基丁酸和多聚磷酸盐的菌株体内含磷量均超过对照菌大肠杆菌,其中GP44的菌体含磷量高达11.92%,本文将对菌株GP44进一步的研究。菌株GP44的个体形态为链杆状,无鞭毛,有荚膜,革兰氏染色阴性,大小为0.75μm×2.25μm,菌落边缘平整,不透明,湿润,呈卵黄色,有臭味。16SrDNA序列分析表明,GP44的16SrDNA基因序列与Klebsiella terrigena ATCC 33257T（AF129442）同源性高达（96%）。通过系统进化树分析和生理生化特征,GP44属于肠杆菌科克雷伯氏菌属中土生克雷伯氏菌伍terrigena)。菌株GP44除磷的最佳培养基为废水合成培养基,生长曲线中菌株的对数生长期在第4h到第16h,在除磷变化曲线中菌株在第24h除磷效率达到最高,此时为测定除磷效率最佳时期。本文研究了非营养因素温度、pH值和溶氧量对菌株GP44除磷效率影响,温度在20-35℃的范围内,菌株的生长对除磷的效果变化不是很大,除磷效果最佳的温度为30℃。在温度为30℃条件下,溶氧量和pH对菌株GP44的除磷效果影响较显著,最佳装液量为120mL·250mL^-1,pH为7.5时生长状况和除磷效果最佳。营养因素碳源、氮源和金属离子对菌株GP44的生长和除磷效果影响的研究表明,最适碳源和氮源分别为为葡萄糖和酵母膏,除磷效率为59.12%;Mg²⁺、K²⁺和Fe³⁺促进菌株GP44的生物除磷,但Fe³⁺对菌株的生长有一定抑制作用,Mn²⁺、Cu²⁺、Pb²⁺、Co²⁺和Zn²⁺对菌株生长和除磷效果均有抑制作用,其他金属离子对菌株的生长和除磷效果影响不明显。正交实验获得最佳除磷效果组合为:以废水合成培养基为基础,葡萄糖0.5g·L^-1,pH值为7.5,装液量为120mL·250mL^-1,培养温度为30℃,四因素对生物除磷效果影响的顺序:溶氧量>pH>碳源>温度。菌株GP44在厌氧好氧循环连续培养时表现出厌氧释磷好氧吸磷的特性,厌氧培养6h释磷量为5.19 mg·L^-1,好氧培养12h吸磷量12.34 mg·L^-1。培养液中的磷浓度对菌株GP44放磷和吸磷有一定的调节作用。溶液中磷浓度为0.89 mg·L^-1时,菌株表现为释放磷,溶液中的磷浓度上升后又呈下降趋势;磷浓度为1.78 mg·L^-1时,菌株表现为摄磷,培养液中的磷浓度呈下降趋势;磷浓度为17.84 mg·L^-1时,菌株的除磷速度和除磷率下降。以城市生活污水为培养基,在厌氧好氧连续2次循环培养后污水中的磷浓度从9.62 mg·L^-1下降为0.86 mg·L^-1,达到国家污水排放二级标准。更多还原

【Abstract】 In this experiment,sixty-six strains were isolated from different sludges by theYG media and thirty strains were fast growing. After incubation in MOPS-glucose agar plates containing Pi-limitaion and Pi-excess respectively for two days. Eleven strains were formed blue-colony on different medium and they were primary PAO. Seven strains could synthesize PHB and polyP in the A/O process. The phosphorus concentraion rate of seven strains exceed E.coil after 24h aerobic incubation and the GP44 reached 11.92% of its weight. This article would do further research about GP44.Microscopic examination showed its shape was bacilliform and this bacterium appeared by short chain, capsule, no flagellum, sporeforming and gram-negative bacteria. The size of the bacteria was 0.75μm×2.25μm approximately. The colony of GP44 is round, smooth margin, opaque, moist, egg-yellow and fishy smell. The 16S rDNA genes of GP44 and Klebsiella terrigena type strain ATCC 33257^T（AF129442） shows a high level of similarity （96%）. According to phenotypic characteristics and phylogenetic analysis, it can be suggested that the strain GP44 has been identified as K. terrigena.Synthesized sewage was the best culture medium for GP44.Logarithmic phase was during four to sixteen hour, the high efficiency of phosphorus removal was at twenty-four hour, and this was the optimum time for tasting the rate of phosphorus removal. In the temperature, pH and aeration experiment, the better temperature range was from 20℃to 40℃,and the best temperature was about 30℃.So influence of temperature is not limited in process of application of PAO to wastewater. But pH and aeration that effected GP44 on growth and phosphorus removal rate were remarkable. The optimal pH value and volume of liquid were 7.5 and 120mL·250mL^-1 respectively. In the carbon source, nitrogen source and metal ion test, glucose and yeast extract were the best carbon source and nitrogen source with high phosphorus removal rate at 59.12%.Mg²⁺、K²⁺and Fe³⁺ could stimulate the phosphorus removal rate of GP44, but Fe³⁺ had restraint to the growth of it.Mn²⁺、Cu²⁺、Pb²⁺、Co²⁺ and Zn²⁺ could restrained the growth and phosphorus removal rate of GP44,and the effect was very remarkable. The others were not. Orthogonal experiment determined the best condition: synthesized sewage as basic medium culture, glucose 0.5g·L^-1,pH7.5, volume of liquid 120mL·250mL^-1 and temperature 30℃,in these four factors, aeration was the main influential factor. The order of phosphorus removal effeced is: aeration > pH> carbon source> temperature. Strain GP44 showed the characteristics of a typical phosphate accumulating organism as it displayed apparent phosphorus accumulated and released in the A/O process. Its release phosphorus was 5.19mg·L^-1whith growth in anaerobic culture for six hours, and accumulated phosphorus was12.34mg·L^-1 in aerobic culture for twelve hours. The phosphorus concentration of medium regulate GP44 to release or accumulate phosphorus to a certain extent. GP44 released phosphorus in the phosphorus concentration of 0.89mg·L^-1,accumulated phosphorus in phosphorus concentrations of 1.78 mg·L^-1,and the phosphorus removal speed and rate were slowly in phosphorus concentration of 17.84 mg·L^-1.The phosphorus concentration in wastewater of 9.62 mg·L^-1to 0.86 mg·L^-1 in the A/O process twice by the municipal wastewater medium. The phosphorus concentration in wastewater is to the twice of Integrated wastewater discharge standard.更多还原