生物滤池及人工湿地净化工厂化海水养殖废水效果研究
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摘要
工厂化循环水养殖模式具有高效、可控、节水省地、对环境污染小等特点,正逐渐在世界各国水产养殖业中得到重视和应用,是未来水产养殖业可持续发展的主要方向之一。本论文以养殖污水处理单元生物滤池和人工湿地为研究核心,进行了封闭循环水养殖系统中生物滤池的挂膜研究及水处理效率分析,并运用PCR-DGGE技术分析了不同时期生物滤池中生物膜上的微生物群落结构变化;模拟养殖废水,构建室内复合垂直流人工湿地系统,探讨了人工湿地净化海水养殖废水的影响因素,在此基础上确定了人工湿地的最佳运行参数,研究了人工湿地净化养殖废水的效果并分析其净化机理,主要结论如下:
1.在生物滤池中添加一定量的微生态制剂和营养盐,经过1个月的生物挂膜,生物载体表面形成了群落结构比较稳定的生物膜。进行封闭式循环水养殖半滑舌鳎生产性试验,系统运行稳定,生物滤池能有效降低水体中NH_4~+-N、NO_2~--N、CODMn和PO_4~(3-)-P的浓度,使养殖水体中污染物的浓度维持在合理的水平,半滑舌鳎生长良好。
2.在生物膜形成过程及系统运行时期,经传统微生物培养的异养细菌,氨氧化细菌,硝化菌的数量都呈现出先增加后逐渐减少直至达到稳定的趋势。异养细菌的数量比氨氧化细菌和硝化菌高出1-2个数量级。各个时期生物载体上微生物群落的组成非常丰富,Shannon指数分别为1.53、1.44、1.57、1.08、1.27和1.30。不同时期生物载体上的微生物种类有一定的变化,存在一些各自特有的种属和共有的种属。除A4期与B1期Cs值偏低外,其余任何两个相邻时期的Cs值都很大,表明微生物群落的变化与演替是缓慢而有规律的,从而保证了生物滤池处理养殖废水效果的稳定性。
3.生物滤池中微生物种群分布较为广泛,鉴定出的33条带分别属于变形菌门(Proteobacteria)的α-、β-、γ-变形菌纲(Proteobacteria) (17个OTUs),厚壁菌门(Firmicutes)的芽孢杆菌纲(Bacilli)(1个OTUs)和拟杆菌门(Bacteroidetes)的黄杆菌纲(Flavobacteria) (15个OTUs),主要优势种群以Proteobacteria和Flavobacteria为主,存在多类细菌共同作用于养殖水体污染物的去除,其中Nitrosomonas参与了系统的硝化过程,参与反硝化过程的主要是黄杆菌(Flavobacteriaceae bacterium)等兼性厌氧细菌。这些功能性细菌在整个系统的运行中起了非常重要的作用。
4.随着盐度的升高,人工湿地对PO_4~(3-)-P的去除率显著下降,但对NH_4~+-N和COD_(Mn)的去除率并没有发生显著的变化,去除率稳定在65%~78%之间,芦苇作为人工湿地植物,经过驯化后有较强的耐盐性,在盐度20下生长良好;随着水力负荷的增加,人工湿地对COD_(Mn)、NH_4~+-N和PO_4~(3-)-P的去除率呈现出降低的趋势,变化显著(P <0.05);当污染负荷增加时,去除率呈现出先增加后降低的趋势,在污染负荷为COD_(Mn) 10 mg/L、NH_4~+-N 2 mg/L、PO_4~(3-)-P 0.5 mg/L时,水质处理效果最高,其平均去除率分别为75%,62%和49%;当DO浓度增加时,人工湿地对COD_(Mn)、NH_4~+-N和PO_4~(3-)-P的去除率无显著变化(P >0.05)。
5.在优化工况下,人工湿地连续运行2个月,SS平均去除率在90%以上,NH_4~+-N和BOD_5平均去除率在70%以上,COD_(Mn)平均去除率在60%以上,NO_2~--N、NO_3~--N、TN、PO_4~(3-)-P、TP、和TOC平均去除率在30%-50%之间,出水水质良好,基本符合养殖用水回用标准。
6.生物滤池和人工湿地在处理和净化海水养殖废水方面具有一定的实际应用价值和优势。
Recirculating aquaculture systems is getting attention and application in the world, and will become one of the main direction of aquaculture industry sustainable development. It has many characteristics, such as efficient, controllable, water-saving and land-saving, little environmental pullution. In this paper, as a core of bio-filter and constructed wetlands, the bio-film formation and efficiency of purifing water of bio-filter were studied; the changes of microbial community structure on bio-carrier of recirculating aquaculture systems during different periods were analysed by PCR-DEEG; simulated mariculture effluent and constructed indoor compound vertical flow constructed wetlands system, the influence factors of constructed wetlands purifying mariculture effluent were discussed, on this base, the optimal operating parameters of the systems were determined, and the systems were operated two months, the performance of treating water and preliminary mechanism were studyed. The main conclusions are as follows:
1. Adding a certain amount of probiotics and nutrient in bio-filter, bio-carrier surface formed community structure relatively stable bio-film after a month of bio-film formation. Then devised productive experiment of breeding Cynoglossus semilaevis Günther using recirculating aquaculture systems. The system run stably, and the bio-filter could reduce the concentration of ammonia-nitrogen, nitrite-nitrogen, chemical oxygen demand and phosphate, which maintained the concentration of pollutants of systems in a reasonable level and Cynoglossus semilaevis Günther grew well.
2. It presented a trend that the quantity of heterotrophic bacteria, ammonia oxidize bacteria and nitrite oxidize bacteria increased gradually to a top and then fallen slowly to a stable level. The number of heterotrophic bacteria was 1-2 orders of magnitude more then ammonium oxidation bacteria and nitrite oxidize. The composition of microbial community of the bio-carrier was very abundant in different periods, and Shannon index was 1.53, 1.44, 1.57, 1.08, 1.27 and 1.30, respectively. During different periods,there was a certain shift in the microbial community structure, which existed some unique species and common species. Beside A4 period and B1 period Cs value(similar index) was low, the Cs value in two adjacent periods was high,indicating the variation and succession of the microbial community was slow and regular, and ensured the stable effect of bio-film treating mariculture effluent.
3. The microbial community distributed widely in bio-filter. The 33 DGGE bands belonged to theα-Proteobacteria,β-Proteobacteria andγ-Proteobacteria of Proteobacteria (seventeen operational taxonomic units), the bacillus of Firmicutes (one operational taxonomic units) and the Flavobacteria of Bacteroidetes (fifteen operational taxonomic units), respectively. Proteobacteria and Flavobacteria were main communities. Several bacteria had an effect on removal of pollutants for farming water and the effluent water quality could meet the requirements of high-density culture. The Nitrosomonas and some other facultative anaerobic bacteria(Flavobacteriaceae bacterium)were identified,which indicated that there may be coexisted pathways of nitrification and denitrification in bio-filter. These functional bacteria played a very important role in operating systems.
4. With the increase of salinity, the removal rate of chemical oxygen demand and ammonia-nitrogen didn’t change significantly (P>0.05), the removal rate of phosphate was decreased gradually and changed significantly (P<0.05), but the reed showed a strong salt-tolerant ability, growing well under the condition when the salinity reached to 20. As the hydraulic load increasing, the removal rate of chemical oxygen demand, ammonia-nitrogen and phosphate also showed a fall tendency and changed greatly (P <0.05). While the pollution load increasing, the removal rate of chemical oxygen demand, ammonia-nitrogen and phosphate were showed a trend that increased first and then decreased gradually. When the pollution was chemical oxygen demand 10 mg﹒L~(-1), ammonia-nitrogen 2 mg﹒L~(-1), phosphate 0.5 mg﹒L~(-1), the efficiency of water treatment was highest, and the average removal rate was 75%, 62%, 49%, respectively. When the concentrations of dissolved oxygen increasing, the removal rate of chemical oxygen demand, ammonia-nitrogen and phosphate had no significant change (p>0.05).
5. In the optimized conditions, the constructed wetlands operated two months. The average removal rate of total suspended solid was more than 90%, the average removal rate of ammonia-nitrogen and biochemical oxygen demand was more than 70%, the average removal rate of chemical oxygen demand was more than 60%, and the average removal rate of nitrite-nitrogen, nitrate-nitrogen, total nitrogen, phosphate, total phosphorous and total organic carbon was between 30% and 50%. The quality of effluent was well, which met the standard of water reuse.
6. Bio-filter and constructed wetlands had a certain practical application value and advantage in purifying mariculture effluent.
1.在生物滤池中添加一定量的微生态制剂和营养盐,经过1个月的生物挂膜,生物载体表面形成了群落结构比较稳定的生物膜。进行封闭式循环水养殖半滑舌鳎生产性试验,系统运行稳定,生物滤池能有效降低水体中NH_4~+-N、NO_2~--N、CODMn和PO_4~(3-)-P的浓度,使养殖水体中污染物的浓度维持在合理的水平,半滑舌鳎生长良好。
2.在生物膜形成过程及系统运行时期,经传统微生物培养的异养细菌,氨氧化细菌,硝化菌的数量都呈现出先增加后逐渐减少直至达到稳定的趋势。异养细菌的数量比氨氧化细菌和硝化菌高出1-2个数量级。各个时期生物载体上微生物群落的组成非常丰富,Shannon指数分别为1.53、1.44、1.57、1.08、1.27和1.30。不同时期生物载体上的微生物种类有一定的变化,存在一些各自特有的种属和共有的种属。除A4期与B1期Cs值偏低外,其余任何两个相邻时期的Cs值都很大,表明微生物群落的变化与演替是缓慢而有规律的,从而保证了生物滤池处理养殖废水效果的稳定性。
3.生物滤池中微生物种群分布较为广泛,鉴定出的33条带分别属于变形菌门(Proteobacteria)的α-、β-、γ-变形菌纲(Proteobacteria) (17个OTUs),厚壁菌门(Firmicutes)的芽孢杆菌纲(Bacilli)(1个OTUs)和拟杆菌门(Bacteroidetes)的黄杆菌纲(Flavobacteria) (15个OTUs),主要优势种群以Proteobacteria和Flavobacteria为主,存在多类细菌共同作用于养殖水体污染物的去除,其中Nitrosomonas参与了系统的硝化过程,参与反硝化过程的主要是黄杆菌(Flavobacteriaceae bacterium)等兼性厌氧细菌。这些功能性细菌在整个系统的运行中起了非常重要的作用。
4.随着盐度的升高,人工湿地对PO_4~(3-)-P的去除率显著下降,但对NH_4~+-N和COD_(Mn)的去除率并没有发生显著的变化,去除率稳定在65%~78%之间,芦苇作为人工湿地植物,经过驯化后有较强的耐盐性,在盐度20下生长良好;随着水力负荷的增加,人工湿地对COD_(Mn)、NH_4~+-N和PO_4~(3-)-P的去除率呈现出降低的趋势,变化显著(P <0.05);当污染负荷增加时,去除率呈现出先增加后降低的趋势,在污染负荷为COD_(Mn) 10 mg/L、NH_4~+-N 2 mg/L、PO_4~(3-)-P 0.5 mg/L时,水质处理效果最高,其平均去除率分别为75%,62%和49%;当DO浓度增加时,人工湿地对COD_(Mn)、NH_4~+-N和PO_4~(3-)-P的去除率无显著变化(P >0.05)。
5.在优化工况下,人工湿地连续运行2个月,SS平均去除率在90%以上,NH_4~+-N和BOD_5平均去除率在70%以上,COD_(Mn)平均去除率在60%以上,NO_2~--N、NO_3~--N、TN、PO_4~(3-)-P、TP、和TOC平均去除率在30%-50%之间,出水水质良好,基本符合养殖用水回用标准。
6.生物滤池和人工湿地在处理和净化海水养殖废水方面具有一定的实际应用价值和优势。
Recirculating aquaculture systems is getting attention and application in the world, and will become one of the main direction of aquaculture industry sustainable development. It has many characteristics, such as efficient, controllable, water-saving and land-saving, little environmental pullution. In this paper, as a core of bio-filter and constructed wetlands, the bio-film formation and efficiency of purifing water of bio-filter were studied; the changes of microbial community structure on bio-carrier of recirculating aquaculture systems during different periods were analysed by PCR-DEEG; simulated mariculture effluent and constructed indoor compound vertical flow constructed wetlands system, the influence factors of constructed wetlands purifying mariculture effluent were discussed, on this base, the optimal operating parameters of the systems were determined, and the systems were operated two months, the performance of treating water and preliminary mechanism were studyed. The main conclusions are as follows:
1. Adding a certain amount of probiotics and nutrient in bio-filter, bio-carrier surface formed community structure relatively stable bio-film after a month of bio-film formation. Then devised productive experiment of breeding Cynoglossus semilaevis Günther using recirculating aquaculture systems. The system run stably, and the bio-filter could reduce the concentration of ammonia-nitrogen, nitrite-nitrogen, chemical oxygen demand and phosphate, which maintained the concentration of pollutants of systems in a reasonable level and Cynoglossus semilaevis Günther grew well.
2. It presented a trend that the quantity of heterotrophic bacteria, ammonia oxidize bacteria and nitrite oxidize bacteria increased gradually to a top and then fallen slowly to a stable level. The number of heterotrophic bacteria was 1-2 orders of magnitude more then ammonium oxidation bacteria and nitrite oxidize. The composition of microbial community of the bio-carrier was very abundant in different periods, and Shannon index was 1.53, 1.44, 1.57, 1.08, 1.27 and 1.30, respectively. During different periods,there was a certain shift in the microbial community structure, which existed some unique species and common species. Beside A4 period and B1 period Cs value(similar index) was low, the Cs value in two adjacent periods was high,indicating the variation and succession of the microbial community was slow and regular, and ensured the stable effect of bio-film treating mariculture effluent.
3. The microbial community distributed widely in bio-filter. The 33 DGGE bands belonged to theα-Proteobacteria,β-Proteobacteria andγ-Proteobacteria of Proteobacteria (seventeen operational taxonomic units), the bacillus of Firmicutes (one operational taxonomic units) and the Flavobacteria of Bacteroidetes (fifteen operational taxonomic units), respectively. Proteobacteria and Flavobacteria were main communities. Several bacteria had an effect on removal of pollutants for farming water and the effluent water quality could meet the requirements of high-density culture. The Nitrosomonas and some other facultative anaerobic bacteria(Flavobacteriaceae bacterium)were identified,which indicated that there may be coexisted pathways of nitrification and denitrification in bio-filter. These functional bacteria played a very important role in operating systems.
4. With the increase of salinity, the removal rate of chemical oxygen demand and ammonia-nitrogen didn’t change significantly (P>0.05), the removal rate of phosphate was decreased gradually and changed significantly (P<0.05), but the reed showed a strong salt-tolerant ability, growing well under the condition when the salinity reached to 20. As the hydraulic load increasing, the removal rate of chemical oxygen demand, ammonia-nitrogen and phosphate also showed a fall tendency and changed greatly (P <0.05). While the pollution load increasing, the removal rate of chemical oxygen demand, ammonia-nitrogen and phosphate were showed a trend that increased first and then decreased gradually. When the pollution was chemical oxygen demand 10 mg﹒L~(-1), ammonia-nitrogen 2 mg﹒L~(-1), phosphate 0.5 mg﹒L~(-1), the efficiency of water treatment was highest, and the average removal rate was 75%, 62%, 49%, respectively. When the concentrations of dissolved oxygen increasing, the removal rate of chemical oxygen demand, ammonia-nitrogen and phosphate had no significant change (p>0.05).
5. In the optimized conditions, the constructed wetlands operated two months. The average removal rate of total suspended solid was more than 90%, the average removal rate of ammonia-nitrogen and biochemical oxygen demand was more than 70%, the average removal rate of chemical oxygen demand was more than 60%, and the average removal rate of nitrite-nitrogen, nitrate-nitrogen, total nitrogen, phosphate, total phosphorous and total organic carbon was between 30% and 50%. The quality of effluent was well, which met the standard of water reuse.
6. Bio-filter and constructed wetlands had a certain practical application value and advantage in purifying mariculture effluent.
引文
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