水源水中隐孢子虫和贾第虫灭活方式及机理研究
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摘要
隐孢子虫(Cryptosporidium)和贾第虫(Giardia)(以下简称“两虫”)可感染人类和其它多种动物,引起发烧、腹痛等症状,对免疫力低下或缺乏的患者甚至导致死亡。“两虫”在水中的存活时间相当长,且对环境因素和传统氯灭菌剂具有较强的抗性,城市常规给水处理工艺很难完全去除“两虫”。我国水厂水处理工艺落后,大多只能对物理和微生物污染进行净化处理,因而具有爆发大规模水源性“两虫”感染的可能,严重威胁广大人民群众的身体健康、生命安全和社会和谐稳定发展。
目前我国对“两虫”的研究仅仅停留在检测方法评估,提高回收率等方面。因此,加大对水中“两虫”杀灭去除的研究已是迫在眉睫,必须尽快解决的大问题。本论文对荧光活性染色法和免疫嘧啶二聚体(TD)荧光检测法进行优化和改进,提高了活性检测的检出率和特异性,考察了水处理杀菌剂和工艺灭活水中“两虫”的效果及相关影响因素,利用扫描电镜(SEM)、蛋白质实验、脂质过氧化氢化反应、聚合酶链式反应(PCR)等技术,对“两虫”灭活机理进行了初步探讨,寻求高效、可行的防控水中“两虫”的方法和工艺,为保障我国城乡饮用水水质质量和居民健康提供强有力的技术支持。
针对水中“两虫”检测检出率低、无法准确评价“两虫”活性的问题,比较了EPA1623法、体外诱导脱囊、改良抗酸染色和荧光活性染色等方法,考察了“两虫”检出率、所用时间等因素,并对荧光活性染色工序进行改进,获得理想的精密度和准确度,证明其是“两虫”活性检测的可靠方法,为进一步研究的开展奠定了基础;
采用荧光活性染色法,通过对典型水处理杀菌剂灭活“两虫”效果及影响因素的考察发现:O_3对“两虫”的灭活能力强于ClO_2,而Cl_2的作用效果最差,达到理想灭活效果时,O_3投加量和作用时间分别为:3.0mg/L和7min;相关影响因素的研究表明,浊度和有机物是影响灭活的主要因素,浊度(有机物浓度)越低,灭活效果越佳,低温条件影响“两虫”的灭活,pH值对“两虫”灭活效果影响较小;
O_3/H_2O_2协同技术具有最佳的灭活“两虫”效果,不仅降低了药剂的初始时浓度(O_3投量2.0mg/L),而且缩短了灭活时间(7.0min),达到理想的“两虫”灭活效果(灭活率99.0%以上)时,H_2O_2/O_3摩尔比为0.8;同时考察了水中常见离子对H_2O_2/O_3灭活效果的影响,低浓度的Ca~(2+),Mg~(2+)和Cu~(2+)等二价金属离子均对灭活起到一定促进作用,一价离子Na~+对灭活无明显的作用,而NO_3~-、HCO_3~-和Cl~-在一定范围内抑制了H_2O_2/O_3对“两虫”的灭活。光催化剂对“两虫”的灭活效果随着催化剂投量的增加,灭活率逐渐增强。当光催化剂为1.0mg/L时,隐孢子虫和贾第虫的灭活率分别为:69.5%和80.0%。随着剂量的增加,灭活率迅速提高,当光催化剂达到5.0mg/L时,隐孢子虫和贾第虫的灭活率分别为:99.8%和99.9%。
以紫外(UV)为代表,考察了物理工艺灭活“两虫”的情况,实验结果表明,UV照射功率越大,灭活能力越强,UV灭活“两虫”的最佳条件为:UV照射功率4.5mW/cm~2,照射剂量13.5mJ/cm~2;浊度和有机物浓度对UV灭活“两虫”影响较为明显,浊度(有机物浓度)提高,UV照射“两虫”后的灭活率相应降低;pH值和温度对UV灭活无明显的促进或抑制。对实际水源水灭活实验表明,UV灭活水中“两虫”具有较强的可行性和一定的应用价值。
以扫描电镜(SEM)、蛋白质实验、脂质过氧化氢化反应和聚合酶链式反应(PCR)等技术,探求各种水处理药剂和工艺处理水中“两虫”过程中的灭活机理,发现:Cl_2和ClO_2灭活“两虫”的机理主要是依靠氧化性和细胞膜穿透能力,致使“两虫”细胞变形,功能缺失,细胞膜通透性增加,从而引起失活或死亡;O_3灭活“两虫”不仅是自身较强的氧化能力,而且和羟基自由基作用密切相关;UV灭活机理在于细胞内核酸生成嘧啶二聚体,甚至引起核酸失活,致使细胞死亡;H_2O_2/O_3协同作用产生大量的羟基自由基对“两虫”的灭活作用最为显著,羟基自由基是最为有效的“两虫”灭活手段。
Cryptosporidium and Giardia (they are referred as two insects) can infect humans and other species of animal, which will cause fever, abdominal pain and other symptoms, and even to cause the death of the patients whose immunity was low or lacked. Two insects can survived in the water as a long time, and they had a strong resistance to environmental factors and chlorine disinfectants, and could not be completely removed by the conventional water treatment process. In China, the water treatment process are slow, and most of the water plants can only purify the physical and microbial contamination, thus, it had the possibility of infection outbreak at largescale, which threat to our country majority of the people's health, safety and development of harmonious and stable society seriously.
At present, the research in our country only concentrated on the development of detection methods, the improved recovery method and so on. So the study on the removal of water beetle is an urgent question in our country. In this study, the fluorescent staining and immune TD (pyrimidine dimer) fluorescence detect methods had been optimizd and improved, and the activity detection rate and specificity of detection had been improved. Besides, the eliminate effect of Fungicide treatment and the processing on two insects. At the same time, the primary investigation of the mechanism of disinfection through scanning electron microscopy (SEM), the protein experiments, lipid peroxidation hydrogenation reaction, polymerase chain reaction (PCR) technology and fungicide treatment. All the work carried out in this paper is to seek effective and feasible prevention and control of water, the two insects, the methods and processes for the protection of urban and rural drinking water quality and health of residents to provide strong technical support.
For the current problem which is poor detection rate and inaccurate evaluate activity, the EPA1623 method, in vitro excystation and modified acid-fast staining methods, such as the detection rate, and establishment of a fluorescent activity staining method were compared. Besides, the "two worm" detection rate, the time and other factors, and the fluorescent activity staining procedure to improve were studied to get a good precision and accuracy, and to imporve that it was the reliable method for "two worms" activity detecting, which was the foundation of a further study to carry out.
Study on the effect and impact factors of inactivation two insects by chemical disinfection, and found that: the inactivation of O_3 to two insects was stronger than that of ClO_2, and Cl_2 had the worst inactivation effect; the best inactivation effect conditions were as follows: the inactivation rate reached more than 99.0%, and the dosage must be above 6.3mg/L with the reaction time was over 360min. Related factors studies have shown that the turbidity and organic matter were the main factors influencing the inactivation, and the lower the turbidity and organic mater were, the better inactivation effect was. Lower temperature was not suitable for the inactivation of two insect, and pH had little effect on the inactivation of the two insects.
In order to achieve the desired inactivation of two insects (inactivation rate above of 99.0%), the two insects inactivated by O_3/H_2O_2 obtained the best results, which not only reduced the initial concentration of bactericide (O_3 dosage of 2.0mg / L), but also shortened the sterilizing time (7.0min). The best H_2O_2/O_3 molar ratio is 0.8. The common ions which influenced inactivation of H_2O_2/O_3 on two insect were investigated in water. Low concentrations of Ca~(2+), Mg~(2+) and Cu~(2+) are divalent metal ions play a catalytic role for the inactivation, Na~+ had no significant effect on disinfection, but NO_3~-, HCO_3~- and Cl~- inhibited H_2O_2/O_3 inactivation of two insects within a certain range. Photocatalytic inactivation of two insects with the amount of catalyst increased the inactivation rate gradually. When the photocatalyst is 1.0 mg/L, the Cryptosporidium and Giardia inactivation rate was 69.5% and 80.0%, respectively. As the dose increased, inactivation rate is rapidly increasing, especially when the photocatalyst was 5.0mg/L, the Cryptosporidium and Giardia inactivation rate reached 99.8% and 99.9% respectively.
Physical inactivation process studies have shown that inactivation of two insects, UV method has a better ability. The higher the UV irradiation power was, the stronger the inactivation capacity was. When the UV radiation power was 4.5mW/cm~2, the irradiation dose increased to 13.5mJ/cm~2, the inactivation rate of the two insects reached 99.9%. Turbidity and the concentration of organic matter on UV inactivation of the "two bugs" is more obvious, when the turbidity (concentration of organic matter) increased, the the inactivation rate of "two worms" after radiated by UV was reduced accordingly. pH value and temperature no significant promote or inhibit effect on the UV inactivation to two insects. The actual water source experimental results show that the UV inactivation of "two worm" has a strong feasibility and value of certain applications.
Scanning electron microscopy (SEM), the protein experiments, lipid peroxidation hydrogenation reaction and polymerase chain reaction (PCR) technology were used to explore a variety of water treatment chemicals and process water treatment on the inactivation process and the mechanism of "two worms". Results showed that the inactivation of "two worm" by Cl_2 and ClO_2 relied mainly on oxidation and cell membrane penetration, resulting in "two worms" cell deformation, which lost function, cell membrane permeability, and then causing inactivation or death. The inactivation of O_3 on two insects was not only relay on its strong oxidizing ability, but also had a colse relationship with the hydroxyl radicals. The mechanism of UV inactivation on two insects was the generation of intracellular nucleic acid pyrimidine dimers, which even lead to inactivation of nucleic acids, and then resulting in cell death. H_2O_2/O_3 synergies produce large amounts of hydroxyl radicals on the inactivation of the two worms, which is the most significant effective method in the inactivation of two worms.
目前我国对“两虫”的研究仅仅停留在检测方法评估,提高回收率等方面。因此,加大对水中“两虫”杀灭去除的研究已是迫在眉睫,必须尽快解决的大问题。本论文对荧光活性染色法和免疫嘧啶二聚体(TD)荧光检测法进行优化和改进,提高了活性检测的检出率和特异性,考察了水处理杀菌剂和工艺灭活水中“两虫”的效果及相关影响因素,利用扫描电镜(SEM)、蛋白质实验、脂质过氧化氢化反应、聚合酶链式反应(PCR)等技术,对“两虫”灭活机理进行了初步探讨,寻求高效、可行的防控水中“两虫”的方法和工艺,为保障我国城乡饮用水水质质量和居民健康提供强有力的技术支持。
针对水中“两虫”检测检出率低、无法准确评价“两虫”活性的问题,比较了EPA1623法、体外诱导脱囊、改良抗酸染色和荧光活性染色等方法,考察了“两虫”检出率、所用时间等因素,并对荧光活性染色工序进行改进,获得理想的精密度和准确度,证明其是“两虫”活性检测的可靠方法,为进一步研究的开展奠定了基础;
采用荧光活性染色法,通过对典型水处理杀菌剂灭活“两虫”效果及影响因素的考察发现:O_3对“两虫”的灭活能力强于ClO_2,而Cl_2的作用效果最差,达到理想灭活效果时,O_3投加量和作用时间分别为:3.0mg/L和7min;相关影响因素的研究表明,浊度和有机物是影响灭活的主要因素,浊度(有机物浓度)越低,灭活效果越佳,低温条件影响“两虫”的灭活,pH值对“两虫”灭活效果影响较小;
O_3/H_2O_2协同技术具有最佳的灭活“两虫”效果,不仅降低了药剂的初始时浓度(O_3投量2.0mg/L),而且缩短了灭活时间(7.0min),达到理想的“两虫”灭活效果(灭活率99.0%以上)时,H_2O_2/O_3摩尔比为0.8;同时考察了水中常见离子对H_2O_2/O_3灭活效果的影响,低浓度的Ca~(2+),Mg~(2+)和Cu~(2+)等二价金属离子均对灭活起到一定促进作用,一价离子Na~+对灭活无明显的作用,而NO_3~-、HCO_3~-和Cl~-在一定范围内抑制了H_2O_2/O_3对“两虫”的灭活。光催化剂对“两虫”的灭活效果随着催化剂投量的增加,灭活率逐渐增强。当光催化剂为1.0mg/L时,隐孢子虫和贾第虫的灭活率分别为:69.5%和80.0%。随着剂量的增加,灭活率迅速提高,当光催化剂达到5.0mg/L时,隐孢子虫和贾第虫的灭活率分别为:99.8%和99.9%。
以紫外(UV)为代表,考察了物理工艺灭活“两虫”的情况,实验结果表明,UV照射功率越大,灭活能力越强,UV灭活“两虫”的最佳条件为:UV照射功率4.5mW/cm~2,照射剂量13.5mJ/cm~2;浊度和有机物浓度对UV灭活“两虫”影响较为明显,浊度(有机物浓度)提高,UV照射“两虫”后的灭活率相应降低;pH值和温度对UV灭活无明显的促进或抑制。对实际水源水灭活实验表明,UV灭活水中“两虫”具有较强的可行性和一定的应用价值。
以扫描电镜(SEM)、蛋白质实验、脂质过氧化氢化反应和聚合酶链式反应(PCR)等技术,探求各种水处理药剂和工艺处理水中“两虫”过程中的灭活机理,发现:Cl_2和ClO_2灭活“两虫”的机理主要是依靠氧化性和细胞膜穿透能力,致使“两虫”细胞变形,功能缺失,细胞膜通透性增加,从而引起失活或死亡;O_3灭活“两虫”不仅是自身较强的氧化能力,而且和羟基自由基作用密切相关;UV灭活机理在于细胞内核酸生成嘧啶二聚体,甚至引起核酸失活,致使细胞死亡;H_2O_2/O_3协同作用产生大量的羟基自由基对“两虫”的灭活作用最为显著,羟基自由基是最为有效的“两虫”灭活手段。
Cryptosporidium and Giardia (they are referred as two insects) can infect humans and other species of animal, which will cause fever, abdominal pain and other symptoms, and even to cause the death of the patients whose immunity was low or lacked. Two insects can survived in the water as a long time, and they had a strong resistance to environmental factors and chlorine disinfectants, and could not be completely removed by the conventional water treatment process. In China, the water treatment process are slow, and most of the water plants can only purify the physical and microbial contamination, thus, it had the possibility of infection outbreak at largescale, which threat to our country majority of the people's health, safety and development of harmonious and stable society seriously.
At present, the research in our country only concentrated on the development of detection methods, the improved recovery method and so on. So the study on the removal of water beetle is an urgent question in our country. In this study, the fluorescent staining and immune TD (pyrimidine dimer) fluorescence detect methods had been optimizd and improved, and the activity detection rate and specificity of detection had been improved. Besides, the eliminate effect of Fungicide treatment and the processing on two insects. At the same time, the primary investigation of the mechanism of disinfection through scanning electron microscopy (SEM), the protein experiments, lipid peroxidation hydrogenation reaction, polymerase chain reaction (PCR) technology and fungicide treatment. All the work carried out in this paper is to seek effective and feasible prevention and control of water, the two insects, the methods and processes for the protection of urban and rural drinking water quality and health of residents to provide strong technical support.
For the current problem which is poor detection rate and inaccurate evaluate activity, the EPA1623 method, in vitro excystation and modified acid-fast staining methods, such as the detection rate, and establishment of a fluorescent activity staining method were compared. Besides, the "two worm" detection rate, the time and other factors, and the fluorescent activity staining procedure to improve were studied to get a good precision and accuracy, and to imporve that it was the reliable method for "two worms" activity detecting, which was the foundation of a further study to carry out.
Study on the effect and impact factors of inactivation two insects by chemical disinfection, and found that: the inactivation of O_3 to two insects was stronger than that of ClO_2, and Cl_2 had the worst inactivation effect; the best inactivation effect conditions were as follows: the inactivation rate reached more than 99.0%, and the dosage must be above 6.3mg/L with the reaction time was over 360min. Related factors studies have shown that the turbidity and organic matter were the main factors influencing the inactivation, and the lower the turbidity and organic mater were, the better inactivation effect was. Lower temperature was not suitable for the inactivation of two insect, and pH had little effect on the inactivation of the two insects.
In order to achieve the desired inactivation of two insects (inactivation rate above of 99.0%), the two insects inactivated by O_3/H_2O_2 obtained the best results, which not only reduced the initial concentration of bactericide (O_3 dosage of 2.0mg / L), but also shortened the sterilizing time (7.0min). The best H_2O_2/O_3 molar ratio is 0.8. The common ions which influenced inactivation of H_2O_2/O_3 on two insect were investigated in water. Low concentrations of Ca~(2+), Mg~(2+) and Cu~(2+) are divalent metal ions play a catalytic role for the inactivation, Na~+ had no significant effect on disinfection, but NO_3~-, HCO_3~- and Cl~- inhibited H_2O_2/O_3 inactivation of two insects within a certain range. Photocatalytic inactivation of two insects with the amount of catalyst increased the inactivation rate gradually. When the photocatalyst is 1.0 mg/L, the Cryptosporidium and Giardia inactivation rate was 69.5% and 80.0%, respectively. As the dose increased, inactivation rate is rapidly increasing, especially when the photocatalyst was 5.0mg/L, the Cryptosporidium and Giardia inactivation rate reached 99.8% and 99.9% respectively.
Physical inactivation process studies have shown that inactivation of two insects, UV method has a better ability. The higher the UV irradiation power was, the stronger the inactivation capacity was. When the UV radiation power was 4.5mW/cm~2, the irradiation dose increased to 13.5mJ/cm~2, the inactivation rate of the two insects reached 99.9%. Turbidity and the concentration of organic matter on UV inactivation of the "two bugs" is more obvious, when the turbidity (concentration of organic matter) increased, the the inactivation rate of "two worms" after radiated by UV was reduced accordingly. pH value and temperature no significant promote or inhibit effect on the UV inactivation to two insects. The actual water source experimental results show that the UV inactivation of "two worm" has a strong feasibility and value of certain applications.
Scanning electron microscopy (SEM), the protein experiments, lipid peroxidation hydrogenation reaction and polymerase chain reaction (PCR) technology were used to explore a variety of water treatment chemicals and process water treatment on the inactivation process and the mechanism of "two worms". Results showed that the inactivation of "two worm" by Cl_2 and ClO_2 relied mainly on oxidation and cell membrane penetration, resulting in "two worms" cell deformation, which lost function, cell membrane permeability, and then causing inactivation or death. The inactivation of O_3 on two insects was not only relay on its strong oxidizing ability, but also had a colse relationship with the hydroxyl radicals. The mechanism of UV inactivation on two insects was the generation of intracellular nucleic acid pyrimidine dimers, which even lead to inactivation of nucleic acids, and then resulting in cell death. H_2O_2/O_3 synergies produce large amounts of hydroxyl radicals on the inactivation of the two worms, which is the most significant effective method in the inactivation of two worms.
引文
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