CN114720586B - 一种评价阴离子对污泥厌氧消化的影响的方法 - Google Patents
一种评价阴离子对污泥厌氧消化的影响的方法 Download PDFInfo
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Abstract
本发明提供一种评价阴离子对污泥厌氧消化的影响的方法,涉及污泥处理领域。该方法包括:将厌氧消化污泥和活性污泥加入到全自动甲烷潜力测试系统的反应器中;向反应器中的污泥加入含有阴离子的溶液作为实验组,以向反应器中的污泥加入等量超纯水作为对照组,以反应器中只加入污泥作为空白组;开启各反应器进行污泥厌氧消化反应直至反应完全;获取如下指标中的至少一种以评价阴离子对污泥厌氧消化的影响:厌氧消化反应产甲烷量、污泥离心后上清液中的阴离子浓度、污泥离心后上清液中的氨氮浓度、厌氧消化反应前后污泥中的微生物多样性水平、厌氧消化反应前后污泥中的微生物组成。
Description
技术领域
本发明涉及污泥处理领域,具体涉及一种评价阴离子对污泥厌氧消化的影响的方法。
背景技术
发明内容
因此,本发明旨在提供一种评价阴离子对污泥厌氧消化的影响的方法。
为实现上述目的,本发明提供如下技术方案:
本发明提供一种评价阴离子对污泥厌氧消化的影响的方法,包括以下步骤:
(1)将厌氧消化污泥和活性污泥加入到全自动甲烷潜力测试系统的反应器中;
(2)向反应器中的污泥加入含有阴离子的溶液作为实验组,以向反应器中的污泥加入等量超纯水作为对照组,以反应器中只加入污泥作为空白组;
(3)开启各反应器进行污泥厌氧消化反应直至反应完全;
(4)获取如下指标中的至少一种以评价阴离子对污泥厌氧消化的影响:厌氧消化反应产甲烷量、污泥离心后上清液中的阴离子浓度、污泥离心后上清液中的氨氮浓度、厌氧消化反应前后污泥中的微生物多样性水平、厌氧消化反应前后污泥中的微生物组成。
进一步地,步骤(1)中,按照厌氧消化污泥与活性污泥的挥发性固体比例为3:2加入所述厌氧消化污泥和活性污泥,所述厌氧消化污泥和活性污泥的总重量为400g。
进一步地,步骤(2)中,含有阴离子的溶液和超纯水的添加量均为5mL,调节反应器内阴离子浓度分别为为2mmol/L、4mmol/L和8mmol/L。
进一步地,步骤(3)中,开启各反应器前向反应器中通入氮气2min;调节反应器搅拌棒转速为100rpm/min,搅拌启动间隔为60s,反应器内温度为37℃,反应器连续运行18天。
进一步地,步骤(3)中,反应过程中向所述全自动甲烷潜力测试系统的气体吸附单元中加入3mol/L的NaOH溶液以吸附污泥厌氧消化反应产生的酸性气体。
进一步地,步骤(4)中,由所述全自动甲烷潜力测试系统的数据处理单元获取厌氧消化反应产甲烷量。
进一步地,步骤(4)中,污泥离心后上清液中的阴离子浓度的检测方法包括:将污泥在4000rpm、4℃条件下离心10min;取上清液,采用C18柱去除有机物和杂质;用超纯水将C18柱过滤后的上清液稀释10倍,经 0.45μm滤膜过滤,采用离子色谱法测定上清液中的浓度。
进一步地,步骤(4)中,污泥离心后上清液中的氨氮浓度的检测方法包括:将污泥在4000rpm、4℃条件下离心10min;取上清液,用超纯水稀释10倍,采用氨气敏电极法测定上清液中的氨氮浓度。
进一步地,步骤(4)中,污泥中的微生物多样性水平通过ACE多样性指数、observedspecies多样性指数、谱系多样性指数、微生物菌群差异性来评价;污泥中的微生物组成通过不同水平上的微生物群落相对丰度来评价。
本发明技术方案,具有如下优点:
本发明提供了一种评价阴离子对污泥厌氧消化的影响的方法,利用全自动甲烷潜力测试系统,通过向污泥中加入含有阴离子的溶液并以厌氧消化反应产甲烷量、污泥离心后上清液中的阴离子浓度和氨氮浓度、厌氧消化反应前后污泥中的微生物多样性水平、微生物组成等作为评价指标,反映出阴离子对污泥厌氧消化的影响,以期对后续污泥处理工作中如何加速污泥厌氧消化、缩短反应器停留时间等提供参考。
附图说明
为了更清楚地说明本发明具体实施方式或现有技术中的技术方案,下面将对具体实施方式或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图是本发明的一些实施方式,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图7是本发明实施例中不同阴离子种类和浓度对反应后污泥上清液中氨氮的影响结果图;
图8是本发明实施例中不同阴离子种类和浓度对反应后污泥中微生物 ACE多样性指数的影响结果图;
图9是本发明实施例中不同阴离子种类和浓度对反应后污泥中微生物 observedspecies多样性指数的影响结果图;
图10是本发明实施例中不同阴离子种类和浓度对反应后污泥中微生物谱系多样性指数的影响结果图;
图11是本发明实施例中不同阴离子种类和浓度对反应后污泥的NMDS 分析图;
图12是本发明实施例中不同阴离子种类和浓度对反应后污泥的PCA 分析图;
图13是本发明实施例中引入不同种类和浓度的阴离子后污泥微生物群落在门水平上的相对丰度图;
图14是本发明实施例中引入不同种类和浓度的阴离子后污泥微生物群落在属水平上的相对丰度图。
具体实施方式
提供下述实施例是为了更好地进一步理解本发明,并不局限于所述最佳实施方式,不对本发明的内容和保护范围构成限制,任何人在本发明的启示下或是将本发明与其他现有技术的特征进行组合而得出的任何与本发明相同或相近似的产品,均落在本发明的保护范围之内。
实施例中未注明具体实验步骤或条件者,按照本领域内的文献所描述的常规实验步骤的操作或条件即可进行。所用原料或仪器,均为可以通过市购获得的常规产品,包括但不限于本申请实施例中采用的原料或仪器。
实施例
1、污泥样品
污泥样品取自北京市的城市污水处理厂,其污水处理工艺为(A2O),污泥处理工艺为厌氧消化。分别从厌氧消化罐与二沉池单元取出厌氧消化污泥与活性污泥样品,在厂内取样后2小时内转移到实验室后避光保存于 4℃环境中。
厌氧消化污泥与活性污泥的主要物理化学特征如表1所示:
表1实验所使用的污泥物理化学指标
样品 | pH | TSS(g/L) | VSS(g/L) |
活性污泥 | 6.56±0.2 | 19.57±0.7 | 17.36±0.4 |
厌氧消化污泥 | 7.68±0.1 | 55.64±0.5 | 24.62±0.3 |
2、试剂
在污泥厌氧消化时引入的阴离子溶液有硝酸钠、亚硝酸钠、氯酸钠和硫酸钠;气体吸附单元使用的氢氧化钠、百里酚酞均采用国药集团生产的分析纯级药品;检测时所需标准溶液(氨氮等)均购自国家标准物质研究中心并进行配制。
3、仪器设备
污泥pH值的测定采用110p-01A STAR pH计(ThermoFisher,Co., USA);污泥厌氧消化反应通过AMPTS-II全自动甲烷潜力测试系统 (Automatic methane potential testsystem II(AMPTS-II)from Bioprocess Control,Sweden)运行;污泥通过高速冷冻离心机(SIGMA 3-16PK)进行离心;离心后污泥上清液中阴离子浓度的测定采用ICS-5000型离子色谱仪 (ThermoFisher,Co.,USA),氨氮浓度的测定采用雷磁PNH3-3氨气敏电极(上海,雷磁),干燥过程采用冷冻干燥机(北京博医康实验仪器有限公司,FD-1A-50)。
AMPTS-II是一种可调整搅拌速率及方向的连续式厌氧反应器,整套系统可分为反应单元(反应器)、气体吸附单元、数据处理单元等。设备运行时,反应器中产生的气体通过连接反应器与气体吸附单元的硅胶软管排出,而后通过气体吸附单元中的氢氧化钠溶液(3mol/L),此时反应器中可能产生的CO2、H2S等酸性气体被氢氧化钠溶液吸附,从而排除其对反应产甲烷量测定的干扰。未被氢氧化钠溶液吸收的气体经硅胶软管排出进入数据处理单元,利用浸泡在去离子水中的气阀记录每组反应器的产甲烷量,最终提供每小时或每天累积产甲烷量、总产甲烷量等信息。
4、操作方法
(1)加入厌氧消化污泥和活性污泥
按照厌氧消化污泥与活性污泥的挥发性固体(VS)比例为3:2将厌氧消化污泥和活性污泥加入到全自动甲烷潜力测试系统的反应器中,厌氧消化污泥和活性污泥的总重量为400g。
(2)加入含有阴离子的溶液
分别向不同的反应器中的污泥加入5mL硝酸钠、亚硝酸钠、氯酸钠或硫酸钠作为实验组,分析引入同一种盐溶液时,改变阴离子引入浓度研究不同浓度阴离子对污泥厌氧消化反应造成的影响,使反应器各内相应阴离子浓度分别为2mmol/L、4mmol/L和8mmol/L;以向反应器中的污泥加入等量超纯水作为对照组;以反应器中只加入污泥作为空白组。
(3)模拟污泥厌氧消化反应
污泥混合完毕后迅速密封,通过硅胶软管向每一反应器中通入氮气 2min,确保反应器中氧气完全排出,使反应器内达到进行厌氧消化反应所需的无氧条件。
开启各反应器进行污泥厌氧消化反应,调节反应器搅拌棒转速为 100rpm/min,搅拌启动间隔为60s,反应器内温度为37℃,反应器连续运行 18天,所有反应器在数据处理单元内显示产甲烷量趋近于0,认为厌氧消化反应产甲烷量已经达到饱和,产气数据不再增加。
(4)获取评价指标
a.产甲烷量
通过全自动甲烷潜力测试系统的数据处理单元读取厌氧消化反应产甲烷量。
反应结束后,取出污泥样品置于冷冻离心机中,在4000rpm、4℃条件下离心10min;取上清液,采用C18柱去除有机物和杂质;用超纯水将C18柱过滤后的上清液稀释10倍,经0.45μm滤膜过滤,采用离子色谱法测定上清液中的浓度。
c.污泥上清液中氨氮浓度
反应结束后,取出污泥样品置于冷冻离心机中,在4000rpm、4℃条件下离心10min;取上清液,用超纯水稀释10倍,采用氨气敏电极法测定上清液中的氨氮浓度。
d.微生物多样性水平、微生物菌群差异性和微生物组成
反应结束后,取出污泥样品进行冷冻干燥,与未进行反应的混合污泥样品(冷冻保存)一并送至上海启因生物科技有限公司进行16S rRNA V3、 V4区高通量测序。通过对测序数据进行处理得到ACE多样性指数、observed species多样性指数、谱系多样性指数、微生物菌群差异性以及不同水平上的微生物群落相对丰度等。
(5)结果分析
a.阴离子种类和浓度对厌氧消化产甲烷量的影响
如图1所示,引入2mmol/L、4mmol/L和8mmol/L NaNO3的累积产甲烷量均高于未引入阴离子的对照组。对照组累积产甲烷量为409.5mL,引入 2mmol/L后产甲烷量为414.2mL;进一步增大投量至4mmol/L,产甲烷量相应提高至422.9mL;继续增大至8mmol/L,产甲烷量不再明显升高。
如图2所示,较低浓度的Na2SO4引入对厌氧消化产气表现出一定的促进作用,而高浓度的Na2SO4引入则抑制厌氧消化产气。引入2mmol/L的时,累积产甲烷量升高至438.7mL,为实验组与对照组中具有最高产甲烷量的一组。但随着引入浓度持续增加至4mmol/L与8mmol/L时,污泥厌氧消化产甲烷量分别为402.3mL和370.8mL,低于对照组产甲烷量,引入8mmol/L时缩短了产甲烷时间,导致第10天污泥停止产生甲烷。
如图3所示,引入2mmol/L的时,累积产甲烷量由对照组累积产甲烷量409.5mL升高至421.8mL;进一步增大至4mmol/L和8mmol/L,累积产甲烷量分别降低至405.8mL和382.4mL。与引入Na2SO4不同,污泥中浓度升高时前期产气速率明显降低。反应开始的前8天时,对照组以及引入2mmol/L、4mmol/L、8mmol/L后的累积产甲烷量分别为 405.1mL、388.2mL、366.8mL和277.46mL。
如图4所示,引入NaClO3对污泥在厌氧消化产气抑制作用,且随着增大抑制效应越明显。具体而言引入为0mmol/L、2mmol/L、 4mmol/L和8mmol/L时,污泥厌氧消化累积产甲烷量分别为409.5mL、 393.3mL、363.9mL、353.0mL。
在上述各种实验条件下,NaClO3投量为8mmol/L时对厌氧消化产气表现出最显著的抑制作用,而Na2SO4为2mmol/L时表现出最明显促进产气作用。
c.阴离子种类和浓度对反应后污泥上清液中氨氮的影响
如图7所示,因为停留时间较长,样品氨氮浓度范围在2.5~9.4g/L。向污泥中引入不同浓度时,氨氮浓度范围依次为 2.5~3.0g/L、3.0~3.5g/L、3.0~5.0g/L和7.0~9.6g/L。污泥中引入的和 浓度增加时,上清液中氨氮浓度升高幅度较小,与对照组氨氮浓度 (2.6g/L)接近;污泥中和浓度增加时,上清液中氨氮浓度升高幅度较大,显著高于对照组氨氮浓度。由于引入并未导致污泥上清液中氨氮浓度增高,所以认为引入的参与了厌氧消化反应,被厌氧污泥中微生物吸收,用于生长与代谢。
结合图1~4和图7,对比不同反应器中氨氮与产甲烷量之间的关系。随着污泥上清液中氨氮浓度的增高,相应的厌氧消化反应中产甲烷量受到了抑制。向厌氧消化反应前引入4mmol/L、8mmol/L的时,污泥上清液中氨氮浓度大于9g/L,与未添加碳源的空白组污泥上清液氨氮浓度9.1g/L 相近。引入后导致污泥中氨氮浓度增加,高浓度氨氮改变了微生物生存条件,是污泥厌氧消化反应产气被抑制的原因。
d.阴离子种类和浓度对污泥微生物多样性的影响
图8~10示出了引入不同种类和浓度阴离子之后污泥微生物的α多样性水平。由图可以看出,引入时ACE、observed species和谱系多样性指数值较高,而的ACE、observed species和谱系多样性指数值的较低;引入阴离子均在不同程度上影响了微生物的α多样性水平。具体而言,往污泥体系中引入不同浓度和时,ACE 值依次为998±54、889±49、872±45和1032±94,observed species值依次为 882±66、761±40、733±26和821±47,而谱系多样性指数依次为94±5、83±4、 83±3、88±3。ACE、observed species、谱系多样性指数等指标的数值越高,说明微生物群落多样性越高。上述结果表明,引入和使得污泥厌氧消化后污泥的微生物种群更丰富,而和则使得污泥中的微生物物种数目更低。
采用β多样性的非度量多维尺度分析(NMDS)和主成分分析(PCA) 方法研究了引入不同种类和浓度的阴离子经厌氧消化后污泥的微生物菌群差异性,结果如图11和12所示。
由图11可以看出,引入8mmol/L时与所有样品之间距离最远,引入2mmol/L、4mmol/L时与其他样品之间距离较远。因此,引入8mmol/L 时与其他组的群落结构具有显著差异,引入2mmol/L、4mmol/L的组与剩余组间在群落结构上存在较大差异。
如图12所示,PCA分析结果显示引入不同阴离子时在PC1轴和PC2 轴上的差异。与NMDS结果相似,引入8mmol/L时与所有样品之间距离最远,引入4mmol/L2mmol/L时与其他样品之间距离较远。但引入2mmol/L、4mmol/L、8mmol/L时距离较小。同NMDS中的分析,引入8mmol/L时微生物群落结构与其他组有明显差异,引入 4mmol/L2mmol/L时与剩余组间在群落结构上存在较大差异。
e.阴离子种类和浓度对污泥微生物群落组成的影响
通过扩增子测序,15个样品共得到了1902个OTUs。
如图13所示,在门的水平上,厚壁菌、变形菌、拟杆菌、互养菌、硝化螺菌、Atribacteria,放线菌、绿弯菌和软壁菌是相对丰度前九的类别。与未反应的污泥相比,厌氧消化反应后污泥的微生物组成均发生了显著变化,主要表现为变形菌和硝化螺菌的相对丰度降低,以及厚壁菌和软壁菌的相对丰度升高。未反应的污泥样品是厌氧消化污泥和活性污泥的混合物,变形菌和硝化相关的菌的相对丰度较高,在厌氧发酵过程中,厚壁菌门作为适应厌氧消化并产生挥发性脂肪酸(VFA)的门类,其相对丰度迅速上升,且软壁菌门相对丰度也升高,而硝化螺菌等由于营养环境的改变而消减。
如图13所示,分析所有培养的污泥样品门水平上的微生物群落结构,得到厚壁菌门、变形菌门、拟杆菌门为厌氧消化后样品中相对丰度占居前三的类别,其相对丰度分别约43.5~77.9%、5.5~23.6%、3.5~12.3%。引入 4mmol/L的污泥中拟杆菌的相对丰度最高,引入8mmol/L时,污泥中变形菌门的相对丰度最高。变形菌门和拟杆菌门的相对丰度在引入 和的样品中分别得到提高,而厚壁菌门相对丰度与对照组相比更低,引入8mmol/L时达到最低。厚壁菌门的相对丰度在引入 2mmol/L时最高,并随着浓度的升高而降低。
进一步分析对照组与引入不同种类和浓度阴离子之后污泥在属水平相对丰度前十的微生物,如图14所示,厚壁菌门的(Fastidiosipila)属、 (Lactobacillus)属和变形菌门的(Arcobacter)是相对丰度前三的属,相对丰度分别约5.2~15.6%、2.0~7.5%和0~7.8%。
引入2mmol/L、4mmol/L、8mmol/L时,Dechloromonas分别为1.8%、 2.2%、3.6%,引入8mmol/L时Thauera的相对丰度为2.5%。 Dechloromonas和Thauera具有反硝化脱氮的能力。因此,污泥中浓度增加提高了污泥反硝化能力。
Fastidiosipila和Lactobacillus能够有机物分解有机酸。污泥引入 2mmol/L、4mmol/L时,Fastidiosipila的相对丰度分别降为4.1%、5.4%、 Lactobacillus的相对丰度引入2mmol/L、4mmol/L时趋于0。明显低于对照组中Fastidiosipila和Lactobacillus的相对丰度。引入2mmol/L、4mmol/L 导致污泥中氨氮浓度增加抑制了Fastidiosipila和Lactobacillus的生长,影响了水解酸化的速度反应,因此厌氧消化反应受到抑制。另外,引入 8mmol/L时虽然未抑制Lactobacillus,但促进了以甲烷为碳源的 Methylobacter;导致引入8mmol/L时,厌氧消化反应产甲烷量最低。
显然,上述实施例仅仅是为清楚地说明所作的举例,而并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (7)
(1)将厌氧消化污泥和活性污泥加入到全自动甲烷潜力测试系统的反应器中;
(2)向反应器中的污泥加入含有阴离子的溶液作为实验组,以向反应器中的污泥加入等量超纯水作为对照组,以反应器中只加入污泥作为空白组;
(3)开启各反应器进行污泥厌氧消化反应直至反应完全,其中,开启各反应器前向反应器中通入氮气2min;调节反应器搅拌棒转速为100rpm/min,搅拌启动间隔为60s,反应器内温度为37℃,反应器连续运行18天;
(4)获取如下指标以评价阴离子对污泥厌氧消化的影响:厌氧消化反应产甲烷量、污泥离心后上清液中的阴离子浓度、污泥离心后上清液中的氨氮浓度、厌氧消化反应前后污泥中的微生物多样性水平、厌氧消化反应前后污泥中的微生物组成,其中,污泥中的微生物多样性水平通过ACE多样性指数、observedspecies多样性指数、谱系多样性指数、微生物菌群差异性来评价;污泥中的微生物组成通过不同水平上的微生物群落相对丰度来评价。
2.根据权利要求1所述的评价阴离子对污泥厌氧消化的影响的方法,其特征在于,步骤(1)中,按照厌氧消化污泥与活性污泥的挥发性固体比例为3:2加入所述厌氧消化污泥和活性污泥,所述厌氧消化污泥和活性污泥的总重量为400g。
3.根据权利要求1所述的评价阴离子对污泥厌氧消化的影响的方法,其特征在于,步骤(2)中,含有阴离子的溶液和超纯水的添加量均为5mL,调节反应器内阴离子浓度分别为2mmol/L、4mmol/L和8mmol/L。
4.根据权利要求1所述的评价阴离子对污泥厌氧消化的影响的方法,其特征在于,步骤(3)中,反应过程中向所述全自动甲烷潜力测试系统的气体吸附单元中加入3mol/L的NaOH溶液以吸附污泥厌氧消化反应产生的酸性气体。
5.根据权利要求1所述的评价阴离子对污泥厌氧消化的影响的方法,其特征在于,步骤(4)中,由所述全自动甲烷潜力测试系统的数据处理单元获取厌氧消化反应产甲烷量。
7.根据权利要求1所述的评价阴离子对污泥厌氧消化的影响的方法,其特征在于,步骤(4)中,污泥离心后上清液中的氨氮浓度的检测方法包括:将污泥在4000rpm、4℃条件下离心10min;取上清液,用超纯水稀释10倍,采用氨气敏电极法测定上清液中的氨氮浓度。
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CN107063922A (zh) * | 2017-02-27 | 2017-08-18 | 湖北洛克泰克仪器有限公司 | 一种全自动甲烷潜力测试系统 |
AU2018241518B2 (en) * | 2017-03-30 | 2023-11-02 | The University Of Queensland | Process for the treatment of sludge |
CN109942159A (zh) * | 2019-03-11 | 2019-06-28 | 广州市香港科大霍英东研究院 | 一种用亚硫酸盐处理剩余污泥的方法 |
CN110330200B (zh) * | 2019-06-28 | 2021-10-22 | 昆明理工大学 | 一种CNTs-Ni-Fe3O4促进厌氧消化过程并提高产甲烷效率的方法 |
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