亲油缓释肥料的制备及在溢油污染潮间带生物修复中的应用
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摘要
生物修复具有二次污染少、费用低廉等优点,已成为现场去除溢油岸滩石油烃污染的重要选择途径。在溢油污染海岸线环境中,营养盐如氮、磷常常是限制石油烃生物降解的主要因素。为了彻底降解并达到更快的净化程度,常常通过添加营养盐即生物刺激的方式来强化污染物的生物降解。
本论文选择环渤海天津海岸线这一溢油高生态风险区,综合采用现场调查、实验室处理和现场模拟修复的综合研究方法,在通过现场调查确定环渤海典型海岸线生态环境特征的基础上,依据石油烃降解菌对不同营养元素吸收利用的特点,开发兼具亲油性和缓释性肥料配方和制备工艺；在室内评价其亲油性、缓释性及对石油烃降解作用影响的基础上,进一步通过现场中试试验,确定所制备的亲油缓释肥料的在不同类型溢油污染海岸线生物修复中的作用过程和作用机理,以期为亲油缓释肥料的开发及其在溢油污染海岸线生物修复中的应用提供理论依据和科学基础。论文主要研究结果如下：
(1)2008年夏季环渤海典型海岸线23个站位的生态环境要素调查结果表明,夏季环渤海的岸线的温度、pH和DO等环境因子基本在石油烃降解菌生长的适宜范围内。环渤海海岸线有机污染较为严重,石油烃和总有机碳含量分别在57.90 mg/Kg-2788.60 mg/Kg和0.15%-6.16%,超过国家沉积物质量Ⅰ类标准的分别占总调查站位的70%和61%。高值区主要位于天津港六号码头、天津汉沽蔡家堡码头、唐山曹妃甸和辽宁营口辽河口；环渤海海岸线富营养化污染较为严重,基质中间隙水总溶解氮和和总溶解磷含量分别在15.75μmol/L-55.70μmol/L范围内和1.10μmol/L-2.17μmol/L,但在溢油发生时,氮、磷营养盐仍可能是限制石油烃生物降解的主要因素。岸线基质中异养菌总数、石油烃降解菌总数、烷烃降解菌总数和芳烃降解菌总数分别在4×104-7.40×106CFU/g、0-8×105CFU/g、170-3.50×106CFU/g和0-1.58×103CFU/g范围内,其中,烷烃降解菌含量较为丰富,而芳烃降解降解菌则普遍缺乏。
(2)通过室内单因子试验,确定了以颗粒状氮肥和磷肥为核心层,以硫磺为包衣层,以聚合蜡为最外密封和涂覆层的亲油缓释肥料的制备工艺,其中,硫包衣层为20%、聚合蜡密封层为4%、表面调理剂为1%、喷涂温度为170℃时肥料的缓释性能最好。该肥料不仅在不同盐度和pH的海水介质中都具有良好的缓释性能,而且能够持续释放养分而维持石油烃降解菌的生长,并促进原油中各组分包括烷烃和芳烃明显降解,其中,各正构烷烃的降解率提高13.9%-59.0%,、平均提高41.2%；芳烃类化合物的降解率提高9.6%-32.3%,平均提高19.9%。
(3)将所制备的亲油缓释肥料单独或与石油降解菌剂同时应用于中质原油和重质原油溢油的现场生物修复中,并与水溶性肥料的作用结果相比较。结果表明,亲油缓释肥料能和水溶性肥料在加入初期都可以大大提高基质间隙水中营养盐的水平,但水溶性肥料在海水冲刷作用下极易流失而导致肥效很短(≤7天),亲油缓释肥料中营养物质释放周期可达25-30天；在释放周期内,亲油缓释肥料能稳定的释放出足够的营养盐为微生物的生长繁殖提供养料,添加亲油缓释肥料的体系各功能菌包括异养菌、石油烃降解菌、烷烃降解菌和芳烃降解菌的数量都明显高于添加水溶性肥料的体系,而各修复体系微生物群落结构的PCR-DGGE分析结果表明,同时添加亲油缓释肥料和接种菌剂的体系的菌群结构明显不同于同时添加水溶性肥料和接种菌剂的体系以及油对照体系,表明所添加缓释肥料不仅可显著刺激土著石油烃降解菌的生长,而且明显促进了所添加的外源石油烃降解菌的生长；无论对BXBT中质原油还是SZ36-1重质原油,同时添加亲油缓释肥料和接种石油烃降解菌剂对提高生物修复效率措施最为有效,不仅提高了烷烃组分的降解率,而且提高了多环芳烃组分的降解率。其中BXBT中质原油在60天时总石油烃的生物降解率提高1.4倍,SZ36-1重质原油120天时的总石油烃的生物降解率提高1.5-2.7倍。
论文研究结果不仅可以为亲油缓释肥料的开发及其在溢油污染海岸线生物修复中的适用性评价提供相关方法和理论基础,而且可为促进石油污染海岸线生物修复技术的完善和大范围推广使用提供支持。
Boremediation, which has many advantages such as less secondary pollution and low-cost, has become the important method to remove petroleum hydrocarbon pollution in oil-spilled shoreline. In the oil-spilled shoreline, nutrients such as nitrogen, phosphorus usually are the main limiting factors to biodegradation. In order to thoroughly degraded petroleum and purified the environment, we often strengthen the biodegradation of pollutants by adding nutrients.
This research chose Tianjin shoreline, which was the area of high incidence of oil spilling, as experimental site to comprehensive study of field investigation, laboratory handling and on-site simulated bioremediation. Based on the field investigation of the typical characteristics of the ecological environment of Bohai Sea coastline, we developed fertilizer which is both lipophilic and slow-released according to the characteristics of petroleum hydrocarbon degrading bacteria on nutrient absorption and utilization of different sustained-release fertilizer. Based on the evaluation of lipophilic efficiency, slow-released efficiency and the impact to bioremediation in laboratory, through the further in-suit test to study the role in the process and mechanism of the slow-released fertilizer in bioremediation of oil-spilled shoreline, in order to seek a theoretical foundation and scientific basis of the development of slow-released fertilizer and its application in the bioremediation of oil-spilled shoreline. The main results of this research are shown as follows:
(1) The investigation result of ecological environment of 23 stations Round Bohai Area in Summer,2008 shows that the environmental factors such as temperature, pH, DO et al Round Bohai Sea are appropriate for the growth of hydrocarbon degrading bacteria. The organic pollution of Round Bohai Sea Area are serious, petroleum hydrocarbons and TOC are between 57.90 mg/Kg-2788.60 mg/Kg and 0.15%-6.16% respectively, there are 70% and 61% of total stations higher the National Sediment Quality Criteria class 1. High value areas are mainly located in Pier 6 of Tianjin Port, Tianjin Hangu Caijiabao Pier, Caofeidian and Liaoning Yingkou Liaohe River Estuary. The eutrophication of Round Bohai Sea Area is also serious, the total dissolved nitrogen (TDN) and total dissolved phosphorus (TDP) in matrix pore water are between 15.75μmol/L-55.70μmol/Land 1.10μmol/L-2.17μmol/L respectively. However, the lack of nitrogen and phosphorus are still the main limiting factors to bioremediation. The number of heterotrophic bacteria, petroleum hydrocarbon degrading bacteria, alkane degrading bacteria and aromatic degrading bacteria are between 4×104-7.40×106CFU/g,0-8×105CFU/g,170-3.50×106CFU/g and 0-1.58×103CFU/g respectively. The alkane degrading bacteria are more abundant, while the number of aromatic degrading bacteria is small.
(2) Through laboratory single factor experiments, we found the optimum conditions and technology of preparing lipophilic and slow-release fertilizer, which make particulate nitrogen and phosphorus as the core layer, sulfur as the coating layer, polymer wax as the outer layer. The fertilizer has the highest slow-released efficiency when the sulfur core layer is 20%, the polymer wax layer is 4%, the surface conditioner is 1%, spraying temperature is 170℃. The fertilizer has good sustained release efficiency not only in different salinity and pH of the seawater, but also can maintain the growth of hydrocarbon degrading bacteria with sustained released nutrients, and promote hydrocarbon degrading bacteria degrading the crude oil components, including alkanes and aromatic hydrocarbons, the degradation rate of n-alkanes increased by 13.9%-59.0%, with an average of 41.2%; the degradation rate of aromatic compounds increased by 9.6%-32.3%, with an average of 19.9%.
(3) Apply the lipophilic slow-released fertilizer in the in-suit oil-spilled bioremediation field of medium crude oil and heavy crude oil, and compare the efficiency with water-soluble fertilizer. The result shows that both slow-released fertilizer and water-soluble fertilizer can greatly improve the level of the nutrients in Matrix pore water. But water-soluble fertilizer are easily washed away because of water erosion and make it has a extremely weak Fertilizer effect (≤7 days), however the slow-released fertilizers has a release cycle of up to 25-30 days. In the release cycle, the fertilizer can release enough nutrients stably to maintain the growth of microbes, The number of heterotrophic bacteria, petroleum hydrocarbon degrading bacteria, alkane degrading bacteria and aromatic degrading bacteria in the system adding slow-release fertilizer are much high than that in the system adding water-soluble, the PCR-DGGE result shows that the microbial community heterotrophic bacteria, petroleum hydrocarbon degrading bacteria, alkane degrading bacteria and aromatic degrading bacteria in the system adding slow-release fertilizer are much high than that in the system adding water-soluble, the PCR-DGGE result shows that the microbial community structure of the system adding slow-released fertilizer and microorganism preparations are significantly different from that adding water-soluble and microorganism preparations and Oil control system, which means that the slow-released fertilizer can not only promote the growth of the indigenous microorganisms but also the growth of exogenous microorganisms. For both medium crude oil and heavy crude oil, the most effective way to Improve the efficiency of bioremediation is adding slow-released fertilizer and microorganism preparations, it can not only improve the biodegradation rate of the alkanes but also aromatic hydrocarbons components. For medium crude oil, the degradation rate in the system adding slow-released fertilizer and microorganism preparations are 1.4 times higher than that in oil controlled system; for heavy crude oil, the degradation rate in the system adding slow-released fertilizer and microorganism preparations are 1.5-2.7 times higher than that in oil controlled system. This paper can provide a theoretical foundation and scientific basis both for the development of slow-released fertilizer and its application in the oil-spilled shoreline; it also can improve the bioremediation technology for oil-spilled shoreline and support the wide range usage of bioremediation.
本论文选择环渤海天津海岸线这一溢油高生态风险区,综合采用现场调查、实验室处理和现场模拟修复的综合研究方法,在通过现场调查确定环渤海典型海岸线生态环境特征的基础上,依据石油烃降解菌对不同营养元素吸收利用的特点,开发兼具亲油性和缓释性肥料配方和制备工艺；在室内评价其亲油性、缓释性及对石油烃降解作用影响的基础上,进一步通过现场中试试验,确定所制备的亲油缓释肥料的在不同类型溢油污染海岸线生物修复中的作用过程和作用机理,以期为亲油缓释肥料的开发及其在溢油污染海岸线生物修复中的应用提供理论依据和科学基础。论文主要研究结果如下：
(1)2008年夏季环渤海典型海岸线23个站位的生态环境要素调查结果表明,夏季环渤海的岸线的温度、pH和DO等环境因子基本在石油烃降解菌生长的适宜范围内。环渤海海岸线有机污染较为严重,石油烃和总有机碳含量分别在57.90 mg/Kg-2788.60 mg/Kg和0.15%-6.16%,超过国家沉积物质量Ⅰ类标准的分别占总调查站位的70%和61%。高值区主要位于天津港六号码头、天津汉沽蔡家堡码头、唐山曹妃甸和辽宁营口辽河口；环渤海海岸线富营养化污染较为严重,基质中间隙水总溶解氮和和总溶解磷含量分别在15.75μmol/L-55.70μmol/L范围内和1.10μmol/L-2.17μmol/L,但在溢油发生时,氮、磷营养盐仍可能是限制石油烃生物降解的主要因素。岸线基质中异养菌总数、石油烃降解菌总数、烷烃降解菌总数和芳烃降解菌总数分别在4×104-7.40×106CFU/g、0-8×105CFU/g、170-3.50×106CFU/g和0-1.58×103CFU/g范围内,其中,烷烃降解菌含量较为丰富,而芳烃降解降解菌则普遍缺乏。
(2)通过室内单因子试验,确定了以颗粒状氮肥和磷肥为核心层,以硫磺为包衣层,以聚合蜡为最外密封和涂覆层的亲油缓释肥料的制备工艺,其中,硫包衣层为20%、聚合蜡密封层为4%、表面调理剂为1%、喷涂温度为170℃时肥料的缓释性能最好。该肥料不仅在不同盐度和pH的海水介质中都具有良好的缓释性能,而且能够持续释放养分而维持石油烃降解菌的生长,并促进原油中各组分包括烷烃和芳烃明显降解,其中,各正构烷烃的降解率提高13.9%-59.0%,、平均提高41.2%；芳烃类化合物的降解率提高9.6%-32.3%,平均提高19.9%。
(3)将所制备的亲油缓释肥料单独或与石油降解菌剂同时应用于中质原油和重质原油溢油的现场生物修复中,并与水溶性肥料的作用结果相比较。结果表明,亲油缓释肥料能和水溶性肥料在加入初期都可以大大提高基质间隙水中营养盐的水平,但水溶性肥料在海水冲刷作用下极易流失而导致肥效很短(≤7天),亲油缓释肥料中营养物质释放周期可达25-30天；在释放周期内,亲油缓释肥料能稳定的释放出足够的营养盐为微生物的生长繁殖提供养料,添加亲油缓释肥料的体系各功能菌包括异养菌、石油烃降解菌、烷烃降解菌和芳烃降解菌的数量都明显高于添加水溶性肥料的体系,而各修复体系微生物群落结构的PCR-DGGE分析结果表明,同时添加亲油缓释肥料和接种菌剂的体系的菌群结构明显不同于同时添加水溶性肥料和接种菌剂的体系以及油对照体系,表明所添加缓释肥料不仅可显著刺激土著石油烃降解菌的生长,而且明显促进了所添加的外源石油烃降解菌的生长；无论对BXBT中质原油还是SZ36-1重质原油,同时添加亲油缓释肥料和接种石油烃降解菌剂对提高生物修复效率措施最为有效,不仅提高了烷烃组分的降解率,而且提高了多环芳烃组分的降解率。其中BXBT中质原油在60天时总石油烃的生物降解率提高1.4倍,SZ36-1重质原油120天时的总石油烃的生物降解率提高1.5-2.7倍。
论文研究结果不仅可以为亲油缓释肥料的开发及其在溢油污染海岸线生物修复中的适用性评价提供相关方法和理论基础,而且可为促进石油污染海岸线生物修复技术的完善和大范围推广使用提供支持。
Boremediation, which has many advantages such as less secondary pollution and low-cost, has become the important method to remove petroleum hydrocarbon pollution in oil-spilled shoreline. In the oil-spilled shoreline, nutrients such as nitrogen, phosphorus usually are the main limiting factors to biodegradation. In order to thoroughly degraded petroleum and purified the environment, we often strengthen the biodegradation of pollutants by adding nutrients.
This research chose Tianjin shoreline, which was the area of high incidence of oil spilling, as experimental site to comprehensive study of field investigation, laboratory handling and on-site simulated bioremediation. Based on the field investigation of the typical characteristics of the ecological environment of Bohai Sea coastline, we developed fertilizer which is both lipophilic and slow-released according to the characteristics of petroleum hydrocarbon degrading bacteria on nutrient absorption and utilization of different sustained-release fertilizer. Based on the evaluation of lipophilic efficiency, slow-released efficiency and the impact to bioremediation in laboratory, through the further in-suit test to study the role in the process and mechanism of the slow-released fertilizer in bioremediation of oil-spilled shoreline, in order to seek a theoretical foundation and scientific basis of the development of slow-released fertilizer and its application in the bioremediation of oil-spilled shoreline. The main results of this research are shown as follows:
(1) The investigation result of ecological environment of 23 stations Round Bohai Area in Summer,2008 shows that the environmental factors such as temperature, pH, DO et al Round Bohai Sea are appropriate for the growth of hydrocarbon degrading bacteria. The organic pollution of Round Bohai Sea Area are serious, petroleum hydrocarbons and TOC are between 57.90 mg/Kg-2788.60 mg/Kg and 0.15%-6.16% respectively, there are 70% and 61% of total stations higher the National Sediment Quality Criteria class 1. High value areas are mainly located in Pier 6 of Tianjin Port, Tianjin Hangu Caijiabao Pier, Caofeidian and Liaoning Yingkou Liaohe River Estuary. The eutrophication of Round Bohai Sea Area is also serious, the total dissolved nitrogen (TDN) and total dissolved phosphorus (TDP) in matrix pore water are between 15.75μmol/L-55.70μmol/Land 1.10μmol/L-2.17μmol/L respectively. However, the lack of nitrogen and phosphorus are still the main limiting factors to bioremediation. The number of heterotrophic bacteria, petroleum hydrocarbon degrading bacteria, alkane degrading bacteria and aromatic degrading bacteria are between 4×104-7.40×106CFU/g,0-8×105CFU/g,170-3.50×106CFU/g and 0-1.58×103CFU/g respectively. The alkane degrading bacteria are more abundant, while the number of aromatic degrading bacteria is small.
(2) Through laboratory single factor experiments, we found the optimum conditions and technology of preparing lipophilic and slow-release fertilizer, which make particulate nitrogen and phosphorus as the core layer, sulfur as the coating layer, polymer wax as the outer layer. The fertilizer has the highest slow-released efficiency when the sulfur core layer is 20%, the polymer wax layer is 4%, the surface conditioner is 1%, spraying temperature is 170℃. The fertilizer has good sustained release efficiency not only in different salinity and pH of the seawater, but also can maintain the growth of hydrocarbon degrading bacteria with sustained released nutrients, and promote hydrocarbon degrading bacteria degrading the crude oil components, including alkanes and aromatic hydrocarbons, the degradation rate of n-alkanes increased by 13.9%-59.0%, with an average of 41.2%; the degradation rate of aromatic compounds increased by 9.6%-32.3%, with an average of 19.9%.
(3) Apply the lipophilic slow-released fertilizer in the in-suit oil-spilled bioremediation field of medium crude oil and heavy crude oil, and compare the efficiency with water-soluble fertilizer. The result shows that both slow-released fertilizer and water-soluble fertilizer can greatly improve the level of the nutrients in Matrix pore water. But water-soluble fertilizer are easily washed away because of water erosion and make it has a extremely weak Fertilizer effect (≤7 days), however the slow-released fertilizers has a release cycle of up to 25-30 days. In the release cycle, the fertilizer can release enough nutrients stably to maintain the growth of microbes, The number of heterotrophic bacteria, petroleum hydrocarbon degrading bacteria, alkane degrading bacteria and aromatic degrading bacteria in the system adding slow-release fertilizer are much high than that in the system adding water-soluble, the PCR-DGGE result shows that the microbial community heterotrophic bacteria, petroleum hydrocarbon degrading bacteria, alkane degrading bacteria and aromatic degrading bacteria in the system adding slow-release fertilizer are much high than that in the system adding water-soluble, the PCR-DGGE result shows that the microbial community structure of the system adding slow-released fertilizer and microorganism preparations are significantly different from that adding water-soluble and microorganism preparations and Oil control system, which means that the slow-released fertilizer can not only promote the growth of the indigenous microorganisms but also the growth of exogenous microorganisms. For both medium crude oil and heavy crude oil, the most effective way to Improve the efficiency of bioremediation is adding slow-released fertilizer and microorganism preparations, it can not only improve the biodegradation rate of the alkanes but also aromatic hydrocarbons components. For medium crude oil, the degradation rate in the system adding slow-released fertilizer and microorganism preparations are 1.4 times higher than that in oil controlled system; for heavy crude oil, the degradation rate in the system adding slow-released fertilizer and microorganism preparations are 1.5-2.7 times higher than that in oil controlled system. This paper can provide a theoretical foundation and scientific basis both for the development of slow-released fertilizer and its application in the oil-spilled shoreline; it also can improve the bioremediation technology for oil-spilled shoreline and support the wide range usage of bioremediation.
引文
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