CN110117558B - 一种培养同步脱氮除硫混合菌群的方法 - Google Patents
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Abstract
本发明公开了一种培养同步脱氮除硫混合菌群的方法,包括:在反应器中接种厌氧氨氧化颗粒污泥,以含氨氮、亚硝氮、硫化物、无机盐、微量元素和碳酸氢钠的模拟废水作为进水,在温度为30~36℃、控制水力停留时间恒定的厌氧条件下运行至稳定;然后采用依次梯度提高进水中硫化物浓度、梯度降低氨氮和亚硝氮浓度、再梯度提高硫化物浓度、最后缩短水力停留时间的方法完成同步脱氮除硫混合菌群的培养。本发明以厌氧氨氧化污泥为单一接种物,通过逐级驯化的方法培养出了同步脱氮除硫的混合菌群,有利于对化工厂等同时含氨氮、亚硝氮和硫化物的废水处理。
Description
技术领域
本发明涉及污泥培养技术领域,具体涉及一种培养同步脱氮除硫混合菌群的方法。
背景技术
氮是水生系统的营养元素,也是生态循环的重要组成部分。而过多的氮素排放则会引发水体富营养化等问题。硫化物由于其恶臭、腐蚀性和毒性,对人体甚至整个生态系统有巨大影响。因此,在废水排放前进行脱氮除硫尤为重要。
与传统的物理化学方法相比,生物技术由于其低能源投入且少化学污泥产生,被称为“环境友好”技术。但目前已有的一些废水生物脱氮除硫工艺通常将硫和氮分开处理,或者只关注于硫化物和亚硝酸盐、或者硫化物和硝酸盐的双组分的去除,而氨盐,亚硝酸盐和硫化物的三者共存问题被忽略。事实上,采矿工厂,垃圾填埋场等的出水中,氨盐,亚硝酸盐和硫化物通常被同时排放。因此,为了简化废水处理程序,提高处理效率,节约能源,在单级工艺中培养出能够脱氮除硫的混合菌群尤为关键。
与传统的生物脱氮相比,厌氧氨氧化工艺可在厌氧环境下,通过厌氧氨氧化菌,利用亚硝酸盐为电子受体,将氨氮直接转化为氮气,其高效脱氮和低能耗性能受到越来越多的关注。脱氮硫杆菌在厌氧条件下以反硝化的方式同时参与硫、氮循环,在氧化硫化物的过程中获得能量,以硝酸盐为电子受体生成氮气。厌氧氨氧化菌和脱氮硫杆菌均为自养型细菌,它们的生长无需外加有机碳源,且在一定条件下可以共存。
因此,若用厌氧氨氧化污泥作为接种污泥,通过逐级驯化的方式培养出脱氮硫杆菌,则可在单级工艺中实现脱氮除硫混合菌群的共存,达到氨氮、亚硝氮和硫化物的同步去除的目的。
发明内容
针对本领域存在的不足之处,本发明提供了一种培养同步脱氮除硫混合菌群的方法,通过将主要含厌氧氨氧化菌的菌群逐渐驯化为含厌氧氨氧化菌和脱氮硫杆菌的混合菌群,达到氨氮、亚硝氮和硫化物的同步去除的目的,简化了废水处理程序,减少了占地面积,运行费用低,适宜处理矿业厂、垃圾填埋场和制药厂等排放的同时含有氮素和硫素的废水。
一种培养同步脱氮除硫混合菌群的方法,包括:
(1)在反应器中接种厌氧氨氧化颗粒污泥,以含氨氮、亚硝氮、硫化物、无机盐、微量元素和碳酸氢钠的模拟废水作为进水,在温度为30~36℃、控制水力停留时间恒定的厌氧条件下运行至稳定;
(2)梯度提高进水中硫化物浓度,每一梯度的运行时间为7~21天,运行至反应器出水总氮或总硫去除率为70%~80%;
(3)梯度降低进水中氨氮和亚硝氮浓度,每一梯度的运行时间为1~21天,运行至反应器出水总氮和总硫去除率均大于80%;
(4)再次梯度提高进水中硫化物浓度,每一梯度的运行时间为7~14天,运行至反应器出水总氮和总硫去除率均不低于80%;
(5)梯度缩短水力停留时间,每一梯度的运行时间为1~14天,运行至反应器出水总氮和总硫去除率均不低于90%,完成同步脱氮除硫混合菌群的培养。
所述步骤(1)和步骤(2)为培养的第一阶段。此阶段主要是在稳定运行的厌氧氨氧化反应器中,逐步提高进水硫化物浓度,使反应器能承受尽可能高浓度的硫化物,运行至脱氮性能降低。
步骤(1)中,优选地,所述的厌氧氨氧化颗粒污泥的挥发性悬浮固体浓度为2~4gL-1,该浓度污泥具有较好的活性和沉降性能。
优选地,所述的氨氮与亚硝氮的质量浓度比为1:(0.9~1.1)。根据厌氧氨氧化反应方程式的系数比,该比例浓度能避免亚硝氮反应不完全而积累,导致抑制反应器性能的现象。
更优选地,所述的氨氮与亚硝氮的质量浓度比为1:1。根据厌氧氨氧化反应方程式的系数比,该比例浓度能避免亚硝氮反应不完全而积累,导致抑制反应器性能的现象,且能尽可能使反应器中氨氮和亚硝氮均反应完全。
优选地,所述进水中总氮浓度为140~560mg L-1。该浓度能避免底物浓度不足,而导致微生物饥饿的现象。所述总氮浓度为氨氮与亚硝氮的浓度之和。
优选地,步骤(1)中,所述进水中的硫化物浓度不大于5mg L-1,避免出水中氨氮和亚硝氮浓度变化过大。过高浓度硫化物的添加会破坏微生物菌群的适应性,导致反应器性能出现较大波动。
优选地,为避免所述的模拟废水在进入反应器前发生化学反应,将模拟废水分为A液和B液,运行过程中,A液和B液采用分开进水的方式。
所述A液含有氨氮、亚硝氮、KH2PO4、CaCl2·2H2O、MgSO4·7H2O、KHCO3和微量元素I溶液、微量元素II溶液。
所述微量元素I溶液含有EDTA和FeSO4。
所述微量元素II溶液含有:EDTA、ZnSO4·7H2O、CoCl2·6H2O、MnCl2·4H2O、CuSO4·5H2O、NaMoO4·2H2O、NiCl2·6H2O和H3BO4。
所述B液含有S2-和NaHCO3。
更优选地,所述A液的组成为:70~280mg L-1氨氮、70~280mg L-1亚硝氮、10mgL- 1KH2PO4、5.6mgL-1CaCl2·2H2O、300mgL-1MgSO4·7H2O、1250mgL-1KHCO3和1.25mL L-1微量元素I溶液、1.25mL L-1微量元素II溶液。
所述微量元素I溶液的组成为:5.00gL-1EDTA和9.14gL-1FeSO4;
所述微量元素II溶液的组成为:15.0gL-1EDTA、0.430gL-1ZnSO4·7H2O、0.240gL- 1CoCl2·6H2O、0.990gL-1MnCl2·4H2O、0.250gL-1CuSO4·5H2O、0.220gL-1NaMoO4·2H2O、0.210gL-1NiCl2·6H2O和0.014gL-1H3BO4;
所述B液的组成为:4~221.5mg L-1S2-和3gL-1NaHCO3。
优选地,步骤(1)中,所述的水力停留时间为3.5~5h,使反应器保持最佳性能。
步骤(2)中,优选地,每次提高后的硫化物浓度为提高前的硫化物浓度的1.5~2.5倍,使反应器能快速适应外界硫化物的添加,避免反应器产生抑制现象。
所述步骤(3)为培养的第二阶段,为反应器性能恢复阶段,为避免亚硝酸盐和硫化物对厌氧氨氧化污泥的协同抑制作用,采用降低进水氨氮和亚硝氮浓度的方式来调节反应器。
步骤(3)中,优选地,每次降低后的氨氮和亚硝氮浓度分别为降低前的氨氮和亚硝氮浓度的1/2~3/4,在保证反应器运行性能稳定的前提下,尽量保持高氮负荷。
所述步骤(4)为培养的第三阶段,反应器性能恢复后,继续进行脱氮硫杆菌的驯化。
优选地,步骤(4)中,每次提高后的硫化物浓度为提高前的硫化物浓度的1.5~2.5倍,使反应器能快速适应外界硫化物的添加,避免反应器产生抑制现象。
优选地,步骤(4)中,硫化物浓度不大于步骤(4)中总氮浓度的80%。,避免硫化物浓度过高,导致反应器抑制。
所述步骤(5)为培养的第四阶段,为性能提升阶段,缩短水力停留时间,可以增大反应器负荷,进而提高反应器性能。
步骤(5)中,所述水力停留时间的缩短梯度为0.5~1h。水力停留时间变化过大会导致微生物无法快速适应环境变化,导致反应器出现运行不稳定,运行性能下降等情况。
本发明与现有技术相比,主要优点包括:本发明以厌氧氨氧化污泥为单一接种物,通过逐级驯化的方法培养出了同步脱氮除硫的混合菌群,有利于对化工厂等同时含氨氮、亚硝氮和硫化物的废水处理。
附图说明
图1为实施例的反应器运行性能图;
图中:NLR代表氮负荷率,NRR代表氮去除速率,NRE代表氮去除效率。
具体实施方式
下面结合附图及具体实施例,进一步阐述本发明。应理解,这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法,通常按照常规条件,或按照制造厂商所建议的条件。
取有效体积为0.8L的上流式厌氧污泥床反应器,接种厌氧氨氧化污泥,厌氧氨氧化污泥的挥发性悬浮固体浓度为2.82g L-1,主导菌为CandidatusKuenenia。
以模拟废水为进水,反应器在厌氧、35±1℃恒温,水力停留时间为4h的条件下运行。为避免模拟废水在进入反应器前发生化学反应,将模拟废水分为A液和B液,运行过程中,A液和B液采用分开进水的方式。A液的组成为:氨氮70~280mg L-1,亚硝氮70~280mg L-1,NaHCO3 840mgL-1,KH2PO4 10mgL-1,CaCl2·2H2O 5.6mgL-1,MgSO4·7H2O 300mgL-1,KHCO31250mgL-1和微量元素I溶液、微量元素II溶液,且氨氮和亚硝氮的质量浓度比始终为1:1;氨氮和亚硝氮分别由(NH4)2SO4和NaNO2提供。
微量元素I溶液组成为:EDTA 5.00gL-1,FeSO49.14gL-1;
微量元素II溶液组成为:EDTA 15.0gL-1,ZnSO4·7H2O 0.430gL-1,CoCl2·6H2O0.240gL-1,MnCl2·4H2O 0.990gL-1,CuSO4·5H2O 0.250gL-1,NaMoO4·2H2O 0.220gL-1,NiCl2·6H2O 0.210gL-1,H3BO40.014gL-1。
B液组成为:S2-4~221.5mg L-1和NaHCO33g L-1。S2-由Na2S·9H2O提供。
反应器稳定运行后开始梯度驯化策略,根据进水中硫化物浓度和硫化物浓度变化梯度,分为三个阶段,其运行时间和反应器运行性能分别见表1和图1,并分别在第30、48、90和126天,从反应器中取出泥样进行高通量测序,样品分别命名为S30,S48,S90和S126。
表1进水基质浓度及运行时间
第一阶段首次向进水中添加4mg L-1硫化物,运行16天后,出水的氨氮,亚硝氮和硝氮浓度波动在5%以内,总氮平均去除率为92%,参见图1。通过计算,此时厌氧氨氧化对脱氮的贡献率为99.5±0.02%,说明此时系统中厌氧氨氧化菌为优势菌。之后分别在第17天、31天、40天时,添加硫化物至8mg L-1、16mg L-1、32mg L-1,其出水的氨氮和亚硝氮浓度均低于20mg L-1,总氮去除率分别为91%,92%和93%,保持在90%以上。在样品S30和S48中,其主导属均为NorankfAnaerolineaceae和CandidatusKuenenia。在第49天,提升硫化物浓度为64mg L-1,运行至第56天时,出水的氨氮和亚硝氮浓度增加到74mg L-1和71mg L-1,总氮去除率降低至72%,继续运行至59天,总氮去除率仍低于80%。此时厌氧氨氧化对脱氮的贡献率降至96.81±0.02%。
第二阶段为反应器性能恢复阶段,为防止反应器的进一步恶化,在第60天降低进水总氮浓度为420mg L-1,运行至第78天,反应器出水的氨氮和亚硝氮浓度依旧保持较高浓度,分别为82mg L-1和67mg L-1,总氮去除率为77%。进一步降低进水总氮浓度为280mg L-1,在第80天出水的氨氮和亚硝氮浓度分别降低为16mg L-1和11mg L-1,总氮去除率恢复至90%。
第三阶段继续提高进水硫化物浓度。在第81天,进水总氮浓度为280mg L-1,硫化物浓度为128mg L-1。运行至第93天,出水的氨氮和亚硝氮浓度分别为32mg L-1和38mg L-1,总氮去除率为83%。此时微生物群落结构发生了明显变化,NorankfAnaerolineaceae和CandidatusKuenenia相对丰度明显降低,而Acinetobacter相对丰度明显增加。第94天进水硫化物浓度为221.5mg L-1,运行至108天,出水的氨氮,亚硝氮和硝氮浓度波动在5%以内,出水的氨氮和亚硝氮浓度分别为13mg L-1和9mg L-1,总氮去除率为90%。此时反硝化对脱氮的贡献率为15%。
第四阶段为性能提升阶段。从第109天开始,逐渐缩短水力停留时间(HRT)。第109天,HRT缩短为3h,运行13天后,总氮去除率为88%。第123天进一步缩短HRT为2.5h,运行至126天,反应器出水的氨氮和亚硝氮浓度均低于10mg L-1,总氮去除率达93%,反硝化对脱氮的贡献率增至26%,且硫化物去除率始终保持95%以上。此时其微生物优势属为Thiobacillus(35.42%),Pseudoxanthomonas(11.61%)和Azoarcus(6.21%),说明经过126天的驯化,培养出了同步脱氮除硫的混合菌群。
此外应理解,在阅读了本发明的上述描述内容之后,本领域技术人员可以对本发明作各种改动或修改,这些等价形式同样落于本申请所附权利要求书所限定的范围。
Claims (7)
1.一种培养同步脱氮除硫混合菌群的方法,包括:
(1)在反应器中接种厌氧氨氧化颗粒污泥,以含氨氮、亚硝氮、硫化物、无机盐、微量元素和碳酸氢钠的模拟废水作为进水,在温度为30~36℃、控制水力停留时间恒定的厌氧条件下运行至稳定;
所述的氨氮与亚硝氮的质量浓度比为1:1;
所述进水中的硫化物浓度不大于5mg L-1;
将模拟废水分为A液和B液分开进水;
所述A液的组成为:70~280mg L-1氨氮、70~280mg L-1亚硝氮、10mg L-1KH2PO4、5.6mgL-1CaCl2·2H2O、300mg L-1MgSO4·7H2O、1250mg L-1KHCO3和1.25mL L-1微量元素I溶液、1.25mL L-1微量元素II溶液;
所述微量元素I溶液的组成为:5.00g L-1EDTA和9.14g L-1FeSO4;
所述微量元素II溶液的组成为:15.0g L-1EDTA、0.430g L-1ZnSO4·7H2O、0.240g L- 1CoCl2·6H2O、0.990g L-1MnCl2·4H2O、0.250g L-1CuSO4·5H2O、0.220g L-1NaMoO4·2H2O、0.210g L-1NiCl2·6H2O和0.014g L-1H3BO4;
所述B液的组成为:4~221.5mg L-1S2-和3g L-1NaHCO3;
(2)梯度提高进水中硫化物浓度,每一梯度的运行时间为7~21天,运行至反应器出水总氮或总硫去除率为70%~80%;
(3)梯度降低进水中氨氮和亚硝氮浓度,每一梯度的运行时间为1~21天,运行至反应器出水总氮和总硫去除率均大于80%;
(4)再次梯度提高进水中硫化物浓度,每一梯度的运行时间为7~14天,运行至反应器出水总氮和总硫去除率均不低于80%;
(5)梯度缩短水力停留时间,每一梯度的运行时间为1~14天,运行至反应器出水总氮和总硫去除率均不低于90%,完成同步脱氮除硫混合菌群的培养。
2.根据权利要求1所述的培养同步脱氮除硫混合菌群的方法,其特征在于,所述的厌氧氨氧化颗粒污泥的挥发性悬浮固体浓度为2~4g L-1。
3.根据权利要求1所述的培养同步脱氮除硫混合菌群的方法,其特征在于,步骤(2)中,每次提高后的硫化物浓度为提高前的硫化物浓度的1.5~2.5倍。
4.根据权利要求1所述的培养同步脱氮除硫混合菌群的方法,其特征在于,步骤(3)中,每次降低后的氨氮和亚硝氮浓度分别为降低前的氨氮和亚硝氮浓度的1/2~3/4。
5.根据权利要求1所述的培养同步脱氮除硫混合菌群的方法,其特征在于,步骤(4)中,每次提高后的硫化物浓度为提高前的硫化物浓度的1.5~2.5倍。
6.根据权利要求1或5所述的培养同步脱氮除硫混合菌群的方法,其特征在于,步骤(4)中,硫化物浓度不大于本步骤中总氮浓度的80%。
7.根据权利要求1所述的培养同步脱氮除硫混合菌群的方法,其特征在于,所述水力停留时间的缩短梯度为0.5~1h。
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