CN110407333B - 一种水平潜流湿地反硝化脱氮增强方法 - Google Patents

一种水平潜流湿地反硝化脱氮增强方法 Download PDF

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CN110407333B
CN110407333B CN201910666462.XA CN201910666462A CN110407333B CN 110407333 B CN110407333 B CN 110407333B CN 201910666462 A CN201910666462 A CN 201910666462A CN 110407333 B CN110407333 B CN 110407333B
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柯凡
刁飞
李文朝
潘继征
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Nanjing Institute of Geography and Limnology of CAS
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Abstract

本发明公开了一种水平潜流湿地反硝化脱氮增强方法,本发明包括菌种驯化、菌种筛选、菌种投放三个处理步骤,首先通过驯化培养来实现好氧反硝化菌的富集,并提高菌种的反硝化优势,得到驯化菌种;同时在得到驯化菌种后,本技术方案中通过对驯化菌种进行多次筛选,去除污泥中的衰老菌种及一些无法进行反硝化作用的菌种,得到真正具有较好的反硝化作用的菌种作为投放菌种;最后利用筛选得出的投放菌种与生物载体相互结合,得到固定化填料;防止菌种流失;本发明工艺操作简单,不仅有效实现了水平潜流湿地的好氧反硝化操作,同时保证了好氧反硝化菌种的生物活性及反硝化性能,增强水平潜流湿地的反硝化。

Description

一种水平潜流湿地反硝化脱氮增强方法
技术领域
本发明涉及湿地治理技术领域,具体是一种水平潜流湿地反硝化脱氮增强方法。
背景技术
在湿地管理运行过程中,污水需要经过微曝气垂直流湿地后进入水平潜流湿地,在这过程中污水的溶氧较高,因此在水平潜流湿地中处于好氧状态,而传统理论认为,反硝化是一个严格的厌缺氧过程,氧会抑制反硝化作用酶,并且在有机质氧化过程中氧是首选的电子受体,从而阻止了硝氮和亚硝氮作为电子受体。
近几年来逐渐有研究人员发现了好氧反硝化菌的存在,但人们在各种不同的环境下如土壤﹑沟渠﹑池塘﹑活性污泥﹑沉积物等分离出了一些好氧反硝化菌,但由于好氧反硝化菌数量少,优势不明显,无法有效完成水平潜流湿地的反硝化脱氮过程。
针对上述情况,我们提供了一种水平潜流湿地反硝化脱氮增强方法,需要提高好氧反硝化菌种的反硝化性能,这是我们亟待解决的问题之一。
发明内容
本发明的目的在于提供一种水平潜流湿地反硝化脱氮增强方法,以解决现有技术中的问题。
为实现上述目的,本发明提供如下技术方案:
一种水平潜流湿地反硝化脱氮增强方法,包括以下步骤:
1)菌种驯化;
2)菌种筛选;
3)菌种投放;
4)结束操作。
本技术方案用于水平潜流湿地,在近几年来逐渐有研究人员发现了好氧反硝化菌的存在,但人们在各种不同的环境下如土壤﹑沟渠﹑池塘﹑活性污泥﹑沉积物等分离出了一些好氧反硝化菌,但由于好氧反硝化菌数量少,优势不明显,因此本技术方案提供了一种用于水平潜流湿地的增强好氧反硝化脱氮的方法。
较优化地,包括以下步骤:
1)菌种驯化:
A.准备碳源、氮源、经富集培养后的活性污泥和培养液,浓缩活性污泥,得到污泥浓缩液,备用;
B.将步骤A准备的培养液投入容器中,再加入污泥浓缩液,调节PH,再加入碳源、氮源,使容器内含碳量为390-410ppm,含氮量约为18-22ppm,碳氮比为C/N=20;
C.小型气泵曝气,使容器内的溶氧量为DO≥5,室温下进行培养,连续培养6-7天,得到驯化菌种;
2)菌种筛选:
a)按比例称取原料,并配制筛选培养基,调节PH至7-7.2,120-125℃灭菌20-30min,冷却备用;
b)从步骤1)驯化后的容器内移取泥水混合液,梯度稀释后涂布在筛选培养基上,培养2-3天,待菌落形成后,挑选培养基周围出现蓝色晕圈的菌落,重复划线,获得初次筛选菌;
c)将初次筛选菌接种于装有液体培养基的锥形瓶,培养取样测定氨氮及总氮去除率,取测定的氨氮及总氮去除水平达到菌种正常去除水平85%以上的菌种,即为复选菌;
3)菌种投放;
a)将复选菌接种于TSA培养基中,在150-160r/min的摇瓶中培养,根据复选菌的生长曲线,当复选菌处于对数生长期时完成培养,得到菌悬液;
b)取步骤a)制备的菌悬液,放入锥形瓶中,随即放入处理好的生物载体,在摇床中培养6-8h进行初挂膜,初挂膜后更换新鲜的培养基,以复选菌对数生长末期为周期在摇床中培养,完成固定化,得到固定化填料;
c)控制湿地环境温度和PH,将步骤b)处理好的固定化填料投放至湿地中,完成反硝化脱氮;
4)结束操作。
较优化地,包括以下步骤:
1)菌种驯化:
A.准备碳源、氮源、经富集培养后的活性污泥和培养液,浓缩活性污泥,得到污泥浓缩液,备用;
B.将步骤A准备的培养液投入容量为15L的容器中,再加入污泥浓缩液,调节PH为6-8,再加入碳源、氮源,使容器内含碳量为390-410ppm,含氮量约为18-22ppm,碳氮比为C/N=20;
C.小型气泵曝气,使容器内的溶氧量为DO≥5,室温下进行培养,以24h为周期投加一次碳源、氮源,其中碳源、氮源的投加量与步骤B中投加量相同,连续培养6-7天,得到驯化菌种;步骤1)中利用曝气培养实现好氧反硝化菌种的富集,同时提高好氧反硝化菌种的反硝化性能,便于后续进行菌种筛选和湿地反硝化处理操作;
2)菌种筛选:
a)按比例称取原料,并配制筛选培养基,调节PH至7-7.2,120-125℃灭菌20-30min,冷却备用;
b)从步骤1)驯化后的容器内移取泥水混合液,梯度稀释后涂布在筛选培养基上,30-35℃下培养2-3天,待菌落形成后,挑选培养基周围出现蓝色晕圈的菌落,重复划线,获得初次筛选菌;步骤b)中通过筛选培养基进行初筛,由于筛选培养基中含有溴百里酚蓝作为指示剂,当PH大于7.6时,该培养基会变成蓝色,而菌种反硝化是生成碱的过程,当筛选培养基中进行反硝化时会引起培养基PH升高,从而使菌落周围出现蓝色晕圈,通过该方案完成好氧反硝化菌种的初步筛选,去除其中的衰老死亡菌种及无法进行反硝化的菌种;
c)将初次筛选菌接种于装有液体培养基的锥形瓶,30-35℃下恒温震荡培养24-48h,取样测定氨氮及总氮去除率,取测定的氨氮及总氮去除水平达到菌种正常去除水平85%以上的菌种,即为复选菌;步骤c)中进行复筛,由于某些菌种在代谢过程中也会生成碱,导致培养基PH升高,因此该步骤中通过复筛进一步来确定所得的菌种具有好氧反硝化性能,去除其掺杂的无法进行好氧反硝化的菌种;并通过测定的氨氮及总氮去除水平,筛选出其中具有优良的好氧反硝化性能的菌种作为投放菌种即复选菌;
3)菌种投放;
a)将复选菌接种于TSA培养基中,在150-160r/min的摇瓶中培养,根据复选菌的生长曲线,当复选菌处于对数生长期时完成培养,得到菌悬液;
b)取步骤a)制备的菌悬液,放入锥形瓶中,随即放入处理好的生物载体,在摇床中培养6-8h进行初挂膜,培养条件为30-32℃、135-145r/min,初挂膜后更换新鲜的培养基,以复选菌对数生长末期为周期在摇床中培养,完成固定化,得到固定化填料;步骤3)进行菌种投放,首先制备复选菌的菌悬液,再利用菌悬液与生物载体相互结合,完成菌种的固定化,制备得到固定化填料;
c)控制湿地环境温度为5-7℃,PH为6.5-7.5,将步骤b)处理好的固定化填料投放至湿地中,完成反硝化脱氮;步骤c)中对湿地环境进行调控,使得好氧反硝化菌种在该环境下快速有效的进行反硝化过程;
4)结束操作。
较优化地,所述步骤1)中,在一个周期内,气泵曝气时间为23.5h,沉降及投放碳源、氮源的时间为0.5h。
较优化地,所述步骤1)中,碳源为甲醇、乙醇、乙酸钠、聚己酸内酯(PCL)、果壳中的一种;氮源为硝酸钾。
较优化地,所述果壳为椰子壳、核桃壳中的一种。
本技术方案中选择甲醇、乙醇、乙酸钠、聚己酸内酯(PCL)、果壳中的一种作为碳源,其中更为优选的方案为椰子壳、核桃壳,完全可以替代乙醇,且原料易得,废物利用,适用于大规模使用,成本低。
较优化地,所述步骤3)的b)步骤中,生物载体为海藻酸钠、菌丝球、聚氨酯泡沫中的一种。
本技术方案中生物载体选择为海藻酸钠、菌丝球、聚氨酯泡沫中的一种,聚氨酯泡沫可通过有机吸附作用将好氧反硝化菌种固定在载体内部和外层,吸附量大,抗冲击能力和抗负荷能力强;海藻酸钠可将好氧反硝化菌种包埋在载体内部,三种原料中最优选的为菌丝球,不仅成本低,易培育,而且具有较好的生物相容性,能够使好氧反硝化菌种有效保持生物活性,吸附性能好。
较优化地,所述步骤2)的a)步骤中,筛选培养基的各种物质的浓度配比如下:包括琼脂20g/L、KNO31g/L、KH2PO41g/L、FeCl2·6H2O0.5g/L、CaCl2·7H2O0.2g/L、MgSO4·7H2O1g/L、琥珀酸钠8.5g/L、溴百里酚蓝(BTB)1mL/L。
较优化地,所述步骤2)的c)步骤中,液体培养基的各种物质的浓度配比如下:包括KNO31g/L、KH2PO41g/L、FeCl2·6H2O0.05g/L、CaCl2·7H2O0.02g/L、MgSO4·7H2O1g/L、琥珀酸钠8.5g/L,120℃下灭菌20-25min,冷却备用。
本技术方案中培养液选择为澄江县污水处理厂的出水。
与现有技术相比,本发明的有益效果是:
本发明包括菌种驯化、菌种筛选、菌种投放三个处理步骤,由于好氧反硝化菌在环境中不是优势菌种,自然界存在较少,目前为止,人们在各种不同的环境下如土壤﹑沟渠﹑池塘﹑活性污泥﹑沉积物等分离出了一些好氧反硝化菌,但由于好氧反硝化菌数量少,优势不明显,所以本技术方案中首先通过驯化培养来实现好氧反硝化菌的富集,并提高菌种的反硝化优势,得到驯化菌种;同时在得到驯化菌种后,本技术方案中通过对驯化菌种进行多次筛选,去除污泥中的衰老菌种及一些无法进行反硝化作用的菌种,得到真正具有较好的反硝化作用的菌种作为投放菌种;最后利用筛选得出的投放菌种与生物载体相互结合,得到固定化填料,这样不仅可以提高好氧反硝化菌种的生化反应速率,还可以对菌种进行保护,防止菌种流失。
本发明设计了一种水平潜流湿地反硝化脱氮增强方法,步骤设计合理,工艺操作简单,不仅有效实现了水平潜流湿地的好氧反硝化操作,同时保证了好氧反硝化菌种的生物活性及反硝化性能,增强水平潜流湿地的反硝化,整个工艺造价成本低,原料易获取,具有较高的实用性和广泛的应用前景。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
通过以下实施例1-3进行水平潜流湿地反硝化脱氮的实验,具体包括以下步骤:
其中液体培养基的各种物质的浓度配比如下:包括KNO31g/L、KH2PO41g/L、FeCl2·6H2O0.05g/L、CaCl2·7H2O0.02g/L、MgSO4·7H2O1g/L、琥珀酸钠8.5g/L,120℃下灭菌20-25min,冷却备用;
筛选培养基的各种物质的浓度配比如下:包括琼脂20g/L、KNO31g/L、KH2PO41g/L、FeCl2·6H2O0.5g/L、CaCl2·7H2O0.2g/L、MgSO4·7H2O1g/L、琥珀酸钠8.5g/L、溴百里酚蓝(BTB)1mL/L。
实施例1:
步骤1:菌种驯化,首先准备碳源、氮源、经富集培养后的活性污泥和培养液,浓缩活性污泥,得到污泥浓缩液,备用;再将步骤A准备的培养液投入容量为15L的容器中,再加入污泥浓缩液,调节PH为6,再加入碳源、氮源,使容器内含碳量为390ppm,含氮量约为19.5ppm,碳氮比为C/N=20;最后小型气泵曝气,使容器内的溶氧量为DO≥5,室温下进行培养,以24h为周期投加一次碳源、氮源,其中碳源、氮源的投加量与步骤B中投加量相同,在一个周期内,气泵曝气时间为23.5h,沉降及投放碳源、氮源的时间为0.5h;连续培养6天,得到驯化菌种;其中碳源为聚己酸内酯(PCL);氮源为硝酸钾;
再进行菌种筛选,按比例称取原料,并配制筛选培养基,调节PH至7,120℃灭菌20min,冷却备用;从驯化后的容器内移取泥水混合液,梯度稀释后涂布在筛选培养基上,30℃下培养2天,待菌落形成后,挑选培养基周围出现蓝色晕圈的菌落,重复划线,获得初次筛选菌;再将初次筛选菌接种于装有液体培养基的锥形瓶,30℃下恒温震荡培养24h,取样测定氨氮及总氮去除率,取测定的氨氮及总氮去除水平达到菌种正常去除水平85%以上的菌种,即为复选菌;
接着进行菌种投放,将复选菌接种于TSA培养基中,在150r/min的摇瓶中培养,根据复选菌的生长曲线,当复选菌处于对数生长期时完成培养,得到菌悬液;取制备的菌悬液,放入锥形瓶中,随即放入处理好的生物载体,在摇床中培养6h进行初挂膜,培养条件为30℃、135r/min,初挂膜后更换新鲜的培养基,以复选菌对数生长末期为周期在摇床中培养,完成固定化,得到固定化填料;再控制湿地环境温度为5℃,PH为6.5,将固定化填料投放至湿地中,完成反硝化脱氮;其中生物载体为海藻酸钠。
实施例2:
步骤1:菌种驯化,首先准备碳源、氮源、经富集培养后的活性污泥和培养液,浓缩活性污泥,得到污泥浓缩液,备用;再将步骤A准备的培养液投入容量为15L的容器中,再加入污泥浓缩液,调节PH为7,再加入碳源、氮源,使容器内含碳量为400ppm,含氮量约为20ppm,碳氮比为C/N=20;最后小型气泵曝气,使容器内的溶氧量为DO≥5,室温下进行培养,以24h为周期投加一次碳源、氮源,其中碳源、氮源的投加量与步骤B中投加量相同,在一个周期内,气泵曝气时间为23.5h,沉降及投放碳源、氮源的时间为0.5h;连续培养6天,得到驯化菌种;其中碳源为椰子壳;氮源为硝酸钾;
再进行菌种筛选,按比例称取原料,并配制筛选培养基,调节PH至7.1,123℃灭菌25min,冷却备用;从驯化后的容器内移取泥水混合液,梯度稀释后涂布在筛选培养基上,33℃下培养2天,待菌落形成后,挑选培养基周围出现蓝色晕圈的菌落,重复划线,获得初次筛选菌;再将初次筛选菌接种于装有液体培养基的锥形瓶,33℃下恒温震荡培养36h,取样测定氨氮及总氮去除率,取测定的氨氮及总氮去除水平达到菌种正常去除水平85%以上的菌种,即为复选菌;
接着进行菌种投放,将复选菌接种于TSA培养基中,在155r/min的摇瓶中培养,根据复选菌的生长曲线,当复选菌处于对数生长期时完成培养,得到菌悬液;取制备的菌悬液,放入锥形瓶中,随即放入处理好的生物载体,在摇床中培养7h进行初挂膜,培养条件为31℃、140r/min,初挂膜后更换新鲜的培养基,以复选菌对数生长末期为周期在摇床中培养,完成固定化,得到固定化填料;再控制湿地环境温度为6℃,PH为7,将固定化填料投放至湿地中,完成反硝化脱氮;其中生物载体为海藻酸钠。
实施例3:
步骤1:菌种驯化,首先准备碳源、氮源、经富集培养后的活性污泥和培养液,浓缩活性污泥,得到污泥浓缩液,备用;再将步骤A准备的培养液投入容量为15L的容器中,再加入污泥浓缩液,调节PH为7,再加入碳源、氮源,使容器内含碳量为410ppm,含氮量约为20.5ppm,碳氮比为C/N=20;最后小型气泵曝气,使容器内的溶氧量为DO≥5,室温下进行培养,以24h为周期投加一次碳源、氮源,其中碳源、氮源的投加量与步骤B中投加量相同,在一个周期内,气泵曝气时间为23.5h,沉降及投放碳源、氮源的时间为0.5h;连续培养6-7天,得到驯化菌种;其中碳源为核桃壳;氮源为硝酸钾;
再进行菌种筛选,按比例称取原料,并配制筛选培养基,调节PH至7.2,125℃灭菌20-30min,冷却备用;从驯化后的容器内移取泥水混合液,梯度稀释后涂布在筛选培养基上,30-35℃下培养3天,待菌落形成后,挑选培养基周围出现蓝色晕圈的菌落,重复划线,获得初次筛选菌;再将初次筛选菌接种于装有液体培养基的锥形瓶,30-35℃下恒温震荡培养48h,取样测定氨氮及总氮去除率,取测定的氨氮及总氮去除水平达到菌种正常去除水平85%以上的菌种,即为复选菌;
接着进行菌种投放,将复选菌接种于TSA培养基中,在160r/min的摇瓶中培养,根据复选菌的生长曲线,当复选菌处于对数生长期时完成培养,得到菌悬液;取制备的菌悬液,放入锥形瓶中,随即放入处理好的生物载体,在摇床中培养8h进行初挂膜,培养条件为32℃、145r/min,初挂膜后更换新鲜的培养基,以复选菌对数生长末期为周期在摇床中培养,完成固定化,得到固定化填料;再控制湿地环境温度为7℃,PH为7.5,将固定化填料投放至湿地中,完成反硝化脱氮;其中生物载体为海藻酸钠。
对比例1:
步骤1:菌种驯化,首先准备碳源、氮源、经富集培养后的活性污泥和培养液,浓缩活性污泥,得到污泥浓缩液,备用;再将步骤A准备的培养液投入容量为15L的容器中,再加入污泥浓缩液,调节PH为7,再加入碳源、氮源,使容器内含碳量为400ppm,含氮量约为20ppm,碳氮比为C/N=20;最后小型气泵曝气,使容器内的溶氧量为DO≥5,室温下进行培养,以24h为周期投加一次碳源、氮源,其中碳源、氮源的投加量与步骤B中投加量相同,在一个周期内,气泵曝气时间为23.5h,沉降及投放碳源、氮源的时间为0.5h;连续培养6天,得到驯化菌种;其中碳源为椰子壳;氮源为硝酸钾;
再进行菌种投放,将驯化菌种接种于TSA培养基中,在155r/min的摇瓶中培养,根据复选菌的生长曲线,当复选菌处于对数生长期时完成培养,得到菌悬液;取制备的菌悬液,放入锥形瓶中,随即放入处理好的生物载体,在摇床中培养7h进行初挂膜,培养条件为31℃、140r/min,初挂膜后更换新鲜的培养基,以复选菌对数生长末期为周期在摇床中培养,完成固定化,得到固定化填料;再控制湿地环境温度为6℃,PH为7,将固定化填料投放至湿地中,完成反硝化脱氮;其中生物载体为海藻酸钠。
对比例2:
步骤1:菌种驯化,首先准备碳源、氮源、经富集培养后的活性污泥和培养液,浓缩活性污泥,得到污泥浓缩液,备用;再将步骤A准备的培养液投入容量为15L的容器中,再加入污泥浓缩液,调节PH为7,再加入碳源、氮源,使容器内含碳量为400ppm,含氮量约为20ppm,碳氮比为C/N=20;最后小型气泵曝气,使容器内的溶氧量为DO≥5,室温下进行培养,以24h为周期投加一次碳源、氮源,其中碳源、氮源的投加量与步骤B中投加量相同,在一个周期内,气泵曝气时间为23.5h,沉降及投放碳源、氮源的时间为0.5h;连续培养6天,得到驯化菌种;其中碳源为椰子壳;氮源为硝酸钾;
再进行菌种筛选,按比例称取原料,并配制筛选培养基,调节PH至7.1,123℃灭菌25min,冷却备用;从驯化后的容器内移取泥水混合液,梯度稀释后涂布在筛选培养基上,33℃下培养2天,待菌落形成后,挑选培养基周围出现蓝色晕圈的菌落,重复划线,获得初次筛选菌;再将初次筛选菌接种于装有液体培养基的锥形瓶,33℃下恒温震荡培养36h,取样测定氨氮及总氮去除率,取测定的氨氮及总氮去除水平达到菌种正常去除水平85%以上的菌种,即为复选菌;
接着进行菌种投放,控制湿地环境温度为6℃,PH为7,将复选菌投放至湿地中,完成反硝化脱氮。
结论:通过对比实施例1-3、对比例1、对比例3的菌种反硝化脱氮效果,可以得到以下结论:
1、本技术方案中通过驯化培养得到了驯化菌种,驯化菌种具有较好的反硝化优势,同时驯化培养过程也实现了好氧反硝化菌种的富集;
2、通过对比实施例1-3与对比例1,可以发现本技术方案中通过对驯化菌种进行多次筛选,去除污泥中的衰老菌种及一些无法进行反硝化作用的菌种,能够得到真正具有较好的反硝化作用的菌种作为投放菌种;
3、通过对比实施例1-3与对比例2,可以发现本技术方案中利用筛选得出的投放菌种与生物载体相互结合,得到固定化填料,不仅可以提高好氧反硝化菌种的生化反应速率,还可以对菌种进行保护,防止菌种流失。
本发明设计了一种水平潜流湿地反硝化脱氮增强方法,步骤设计合理,工艺操作简单,不仅有效实现了水平潜流湿地的好氧反硝化操作,同时保证了好氧反硝化菌种的生物活性及反硝化性能,增强水平潜流湿地的反硝化,整个工艺造价成本低,原料易获取,具有较高的实用性和广泛的应用前景。
对于本领域技术人员而言,显然本发明不限于上述示范性实施例的细节,而且在不背离本发明的精神或基本特征的情况下,能够以其他的具体形式实现本发明。因此,无论从哪一点来看,均应将实施例看作是示范性的,而且是非限制性的,本发明的范围由所附权利要求而不是上述说明限定,因此旨在将落在权利要求的等同要件的含义和范围内的所有变化囊括在本发明内。

Claims (4)

1.一种水平潜流湿地反硝化脱氮增强方法,其特征在于:包括以下步骤:
1)菌种驯化:
A.准备碳源、氮源、经富集培养后的活性污泥和培养液,浓缩活性污泥,得到污泥浓缩液,备用;
B.将步骤A准备的培养液投入容量为15L的容器中,再加入污泥浓缩液,调节pH为6-8,再加入碳源、氮源,使容器内含碳量为390-410ppm,含氮量为18-22ppm,碳氮比为C/N=20;
C.小型气泵曝气,使容器内的溶氧量为DO≥5,室温下进行培养,以24h为周期投加一次碳源、氮源,其中碳源、氮源的投加量与步骤B中投加量相同,连续培养6-7天,得到驯化菌种;
2)菌种筛选:
a)按比例称取原料,并配制筛选培养基,调节pH至7-7.2,120-125℃灭菌20-30min,冷却备用;
b)从步骤1)驯化后的容器内移取泥水混合液,梯度稀释后涂布在筛选培养基上,30-35℃下培养2-3天,待菌落形成后,挑选培养基周围出现蓝色晕圈的菌落,重复划线,获得初次筛选菌;
c)将初次筛选菌接种于装有液体培养基的锥形瓶,30-35℃下恒温震荡培养24-48h,取样测定氨氮及总氮去除率,取测定的氨氮及总氮去除水平达到85%以上的菌种,即为复选菌;
3)菌种投放;
a)将复选菌接种于TSA培养基中,在150-160r/min的摇瓶中培养,根据复选菌的生长曲线,当复选菌处于对数生长期时完成培养,得到菌悬液;
b)取步骤a)制备的菌悬液,放入锥形瓶中,随即放入处理好的生物载体,在摇床中培养6-8h进行初挂膜,培养条件为30-32℃、135-145r/min,初挂膜后更换新鲜的培养基,以复选菌对数生长末期为周期在摇床中培养,完成固定化,得到固定化填料;
c)控制湿地环境温度为5-7℃,pH为6.5-7.5,将步骤b)处理好的固定化填料投放至湿地中,完成反硝化脱氮;
4)结束操作;
所述步骤1)中,在一个周期内,气泵曝气时间为23.5h,沉降及投放碳源、氮源的时间为0.5h;
所述步骤1)中,碳源为果壳;氮源为硝酸钾;所述果壳为椰子壳、核桃壳中的一种。
2.根据权利要求1所述的一种水平潜流湿地反硝化脱氮增强方法,其特征在于:所述步骤3)的b)步骤中,生物载体为海藻酸钠、菌丝球、聚氨酯泡沫中的一种。
3.根据权利要求1所述的一种水平潜流湿地反硝化脱氮增强方法,其特征在于:所述步骤2)的a)步骤中,所述筛选培养基的各种物质的浓度配比如下:包括琼脂20g/L、KNO31g/L、KH2PO41g/L、FeCl2·6H2O0.5g/L、CaCl2·7H2O0.2g/L、MgSO4·7H2O1g/L、琥珀酸钠8.5g/L、溴百里酚蓝1mL/L。
4.根据权利要求1所述的一种水平潜流湿地反硝化脱氮增强方法,其特征在于:所述步骤2)的c)步骤中,所述液体培养基的各种物质的浓度配比如下:包括KNO31g/L、KH2PO41g/L、FeCl2·6H2O0.05g/L、CaCl2·7H2O0.02g/L、MgSO4·7H2O1g/L、琥珀酸钠8.5g/L,120℃下灭菌20-25min,冷却备用。
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