外源活性污泥胞外聚合物对小球衣藻聚集行为的影响
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摘要
微藻聚集行为在藻华的治理等方面具有重要作用,也是目前微藻生物燃料收集的瓶颈之一.以小球衣藻为研究对象,观察污泥胞外聚合物(Extracellular polymeric substance,EPS)对微藻聚集行为的影响,并通过其聚集数据采集、EPS客观特性和微藻的自身响应等方面探究其可能的聚集原因.结果显示:随着外源性污泥EPS浓度的增大,小球衣藻聚集率从18.4%上升到81.8%,并且在110 mg/L时达到聚集率最大值.微藻细胞运动速度从26.8μm/s上升到49.4μm/s,增大了细胞间的碰撞几率,促进了微藻的聚集.三维荧光光谱表明,污泥胞外聚合物刺激了微藻色氨酸类蛋白和芳香族类蛋白的分泌,其分泌量与EPS的浓度成正比.但是外源性EPS溶液Zeta电位变化与微藻的聚集率呈负相关关系,并非微藻聚集原因.本研究表明外源性污泥胞外聚合物能够促进微藻的聚集,结果可为下一步在分子方面的探究提供实验基础.(图7表3参38)
Though extracellular polymeric substances(EPSs) play important roles in the aggregation of activated microbial sludge, little is known about their effects on the aggregation of other, non-bacterial species. In this study, the interaction between activated sludge-extracted EPSs and microalgal cells(Chlamydomonas microsphaera) was investigated. Image and video analyses were applied to track microalgal aggregation and motility. The characteristics of the EPSs used and the selfresponses of the tested microalgae were also measured. Results showed that with the application of increasing amounts of exogenous EPSs, the aggregation rate of Chlamydomonas microsphaera increased from 18.4 to 81.8%, with the maximum aggregation rate reached with the application of 110 mg/L of EPSs. In the presence of activated sludge-extracted EPSs,microalgal cells moved faster, with their speed increasing from 26.8 to 49.4 μm/s, which caused there to be higher probabilities of collision among microalgal cells that favored microalgal aggregation. The results of EEM also showed that the EPS content of the activated sludge was proportional to that of the aromatic and tryptophan proteins secreted by the microalgae, which implied that the sludge-extracted EPSs stimulated the secretion of aromatic and tryptophan proteins by the microalgae.However, the zeta potential of the exogenous EPS solution was negatively correlated with the aggregation rate of the microalgae, indicating that zeta potential changes were not responsible for the accumulation of microalgal cells. In conclusion,it was found that sludge-extracted EPSs enhanced the aggregation of microalgae.
Though extracellular polymeric substances(EPSs) play important roles in the aggregation of activated microbial sludge, little is known about their effects on the aggregation of other, non-bacterial species. In this study, the interaction between activated sludge-extracted EPSs and microalgal cells(Chlamydomonas microsphaera) was investigated. Image and video analyses were applied to track microalgal aggregation and motility. The characteristics of the EPSs used and the selfresponses of the tested microalgae were also measured. Results showed that with the application of increasing amounts of exogenous EPSs, the aggregation rate of Chlamydomonas microsphaera increased from 18.4 to 81.8%, with the maximum aggregation rate reached with the application of 110 mg/L of EPSs. In the presence of activated sludge-extracted EPSs,microalgal cells moved faster, with their speed increasing from 26.8 to 49.4 μm/s, which caused there to be higher probabilities of collision among microalgal cells that favored microalgal aggregation. The results of EEM also showed that the EPS content of the activated sludge was proportional to that of the aromatic and tryptophan proteins secreted by the microalgae, which implied that the sludge-extracted EPSs stimulated the secretion of aromatic and tryptophan proteins by the microalgae.However, the zeta potential of the exogenous EPS solution was negatively correlated with the aggregation rate of the microalgae, indicating that zeta potential changes were not responsible for the accumulation of microalgal cells. In conclusion,it was found that sludge-extracted EPSs enhanced the aggregation of microalgae.
引文
1 Xu L,Huo M,Sun C,Cui X,Zhou D,Crittenden JC,Yang W.Bioresources inner-recycling between bioflocculation of Microcystis aeruginosa and its reutilization as a substrate for bioflocculant production[J].Sci Rep,2017,7:43784
2 Laughinghouse HD,Pra D,Silva-Stenico ME,Rieger A,Frescura VDS,Fiore MF,Tedesco SB.Biomonitoring genotoxicity and cytotoxicity of Microcystis aeruginosa(Chroococcales,Cyanobacteria)using the Allium cepa test[J].Sci Total Environ,2012,432(16):180-188
3 Zhang H,Yu Z,Huang Q,Xiao X,Wang X,Zhang F,Wang X,Liu Y,Hu C.Isolation,identification and characterization of phytoplankton-lytic bacterium CH-22 against Microcystis aeruginosa[J].Limnology,2011.41(1):70-77
4 Lee J,Cho DH,Ramanan R,Kim BH,Oh HM,Kim HS.Microalgaeassociated bacteria play a key role in the flocculation of Chlorella vulgaris[J].Bioresour Technol,2013,131(2):195-201
5 Chen CY,Yeh KL,Aisyah R,Lee DJ,Chang JS.Cultivation,photobioreactor design and harvesting of microalgae for biodiesel production:a critical review[J].Bioresour Technol,2011,102(1):71-81
6 Powell RJ,Hill RT.Rapid aggregation of biofuel-producing algae by the bacterium Bacillus sp.strain RP1137[J].Appl Environ Microbiol,2013,79(19):6093
7张佳琪,庞宁,杨舒棋,林泓,胡筱敏,姜彬慧.絮凝法采收生物燃料微藻的研究进展[J].环境保护与循环经济,2017,37(3):25-30[Zhang JQ,Pang N,Yang SQ,Lin H,Hu YM,Jiang BH.Advances in research on harvesting biofuel microalgae by flocculation[J].Liaoning Urban Rural Environ Sci Technol,2017,37(3):25-3 0]
8 Christenson L,Sims R.Production and harvesting of microalgae forwastewater treatment,biofuels,and bioproducts[J].Biotechnol Adv2011,29(6):686-702
9 Uduman N,Qi Y,Danquah MK,Forde GM,Hoadley A.Dewatering of microalgal cultures:a major bottleneck to algae-based fuels[J].J Renew Sustain Energ,2010,2(1):23-571
10 Alam MA,Wan C,Guo SL,Zhao XQ,Huang ZY,Yang YL,Chang JS,Bai FW.Characterization of the flocculating agent from the spontaneously flocculating microalga Chlorella vulgaris JSC-7[J].J Biosc Bioeng,2014,1 18(1):29-33
11 McSwain BS,Irvine RL.Composition and distribution of extracellular polymeric substances in aerobic flocs and granular sludge[J].Appl Environ Microbiol,2005,71(2):1051-1057
12 Liu XM,Sheng GP,Luo HW,Zhang F,Yuan SJ,Xu J,Zeng Raymond J;Wu JG,Yu HQ.Contribution of extracellular polymeric substances(EPS)to the sludge aggregation[J].Environ Sci Technol,2010,44(11):4355-4360
13 Sanin FD,Vesilind PA.Effect of centrifugation on the removal of extracellular polymers and physical properties of activated sludge[J].Water Sci Technol,1994,30(8):117-127
14 Yu GH,He PJ,Shao LM.Characteristics of extracellular polymeric substances(EPS)fractions from excess sludges and their effects on bioflocculability[J].Bioresour Technol,2009,100(13):3193-3 198
15 Wilen BM,Jin B,Lant P.The influence of key chemical constituents in activated sludge on surface and flocculating properties[J].Water Res,2003,37(9):2127-2139
16龙向宇,龙腾锐,唐然,李金印,罗太忠.阳离子交换树脂提取活性污泥胞外聚合物的研究[J].中国给水排水,2008,24(3):29-33[Long XY,Long TR,Tang R,Li JY,Luo TZ.Study on extraction of extracellular polymeric substance from activated sludge using cation exchange resin[J].Chin Wa ter Wa stewater,2008,24(3):29-33]
17 Oh H,Lee S,Park M,Kim H.Harvesting of Chlorella vulgaris using a bioflocculant from Paenibacillus sp.AM49[J].Biotechnol Lett,2001,23(15):1229-1234
18 Xu H,Yu G,Jiang H.Investigation on extracellular polymeric substances from mucilaginous cyanobacterial blooms in eutrophic freshwater lakes[J].Chemosphere,2013,93(1):75-81
19 Chen X,Zhang C,Tan L,Wang J.Toxicity of Co nanoparticles on three species of marine microalgae[J].Environ Pollut,2018,236:454-461
20 Chen W,Westerhoff P,Leenheer JA,Booksh K.Fluorescence excitationemission matrix regional integration to quantify spectra for dissolved organic matter[J].Environ Sci Technol,2003,37(24):5701-5710
21 Chabalina LD,Pastor MR,Rico DP.Characterization of soluble and bound EPS obtained from 2 submerged membrane bioreactors by3D-EEM and HPSEC[J].Talanta,2013,115(17):706-712
22 Liu X,Wu S,Zhang D,Shen J,Han W,Sun X,Li J,Wang L.Simultaneous pyridine biodegradation and nitrogen removal in an aerobic granular system[J].J Environ Sci,2018,67(5):318-329
23 Jia F,Yang Q,Liu X,Li X,Li B,Zhang L,Peng,Y.Stratification of extracellular polymeric substances(EPS)for aggregated anammox microorganisms[J].Environ Sci Technol,2017,51(6):3260-3268
24 Hori K,Matsumoto S.Bacterial adhesion:from mechanism to control[J].Biochem Eng J,2010,48(3):424-434
25 Marshall KC,Stout R,Mitchell R.Mechanism of the initial events in thesorption of marine bacteria to surfaces[J].J Gen Microbiol,1971,68(3):337-348
26张兰河,李军,郭静波,贾艳萍,张海丰.EPS对活性污泥絮凝沉降性能与表面性质的影响[J].化工学报,2012,63(6):1865-1871[Zhang L,Li J,Guo J,Jia Y,Zhang H.Effect of EPS on flocculation-sedimentation and surface properties of activated sludge[J].J Chem Ind Eng(Chin).2012,63(6):1865-1871]
27 Kolter R,Greenberg EP.Microbial sciences:the superficial life of microbes[J].Nature,2006,441(7091):300-302
28 Long Z,Quaife B,Salman H,Oltvai ZN.Cell-cell communication enhances bacterial chemotaxis toward external attractants[J].Sci Rep.2017,7(1)
29 Golzarijalal M,Ashtiani FZ,Dabir B.Modeling of microalgal shearinduced flocculation and sedimentation using a coupled CFD-population balance approach[J].Biotechnol Prog,2018,34(1):160-174
30胡沅胜,刘斌,郝晓地,曹亚莉.微藻处理污水中的絮凝分离/采收研究现状与展望[J].环境科学学报,2015,35(1):12-29[Hu YS,Liu B,Hao XD,Cao YL.Current status and outlook of microalgae flocculation in wastewater treatment[J].Acta Sci Circumt,2015,35(1):12-29]
31 Salim S,Bosma R,Vermue MH,Wijffels.Harvesting of microalgae by bio-flocculation[J].J Appl Phycol,2011,23(5):849-855
32 Ozkan A,Berberoglu H.Adhesion of algal cells to surfaces[J].Biofouling,2013,29(4):469-482
33 Zhou D,Zhang C,Fu L,Xu L,Cui X,Li Q,Crittenden J C.Responses of the microalga Chlorophyta sp.to bacterial quorum sensing molecules(N-Acylhomoserine Lactones):aromatic protein-induced self-aggregation[J].Environ Sci Technol,2017,51(6):3490-3498
34 Ding A,Pronk W,Qu FS,Ma J,Li GB,Li K,Liang H.Effect of calcium addition on sludge properties and membrane fouling potential of the membrane-coupled expanded granular sludge bed process[J].J Membr Sci,2015,489:55-63
35 Meng FG,Zhou ZB,Bing-Jie Ni,Zheng X,Huang GC,Jia XS,Li SY,Xiong Y,Kraume M.Characterization of the size-fractionated biomacromolecules:tracking their role and fate in a membrane bioreactor[J].Wa ter Res,2011,45(15):4661-4671
36宿程远,郑鹏,卢宇翔,谢莲,黄纯萍,黄智,陈孟林.海泡石与生物质炭强化厌氧处理养猪废水[J].中国环境科学,2017,37(10):3764-3772[Su CY,Zheng P,Lu YX,Xie L,Huang C, Huang Z,Chen ML.Enhanced efficiency of an anaerobic reactor containing sepiolite or biochar for treatment swine wastewater[J].Chin Environ Sci,2017,37(10):3764-3772]
37王硕,于水利,徐巧,付强,李激.好氧颗粒污泥特性、应用及形成机理研究进展[J].应用与环境生物学报,2014,20(4):732-742[Wang S,Yu SL,Xu Q,Fu Q,Li J.Characteristics,application and formation mechanisms of aerobic granular sludge:recent advances[J].Chin J Appl Environ Biol,2014,20(4):732-742]
38唐堂,王硕,蒋志坚,李激.群感效应与信号分子在污泥颗粒化过程中的作用研究进展[J].应用与环境生物学报,2016,22(4):718-724[Tang T,Wang S,Jiang ZJ,Li J.Progress in study of the function of quorum sensing and cell signaling in the formation of aerobic granular sludge[J].Chin J Appl Environ Biol,2016,22(4):718-724]
2 Laughinghouse HD,Pra D,Silva-Stenico ME,Rieger A,Frescura VDS,Fiore MF,Tedesco SB.Biomonitoring genotoxicity and cytotoxicity of Microcystis aeruginosa(Chroococcales,Cyanobacteria)using the Allium cepa test[J].Sci Total Environ,2012,432(16):180-188
3 Zhang H,Yu Z,Huang Q,Xiao X,Wang X,Zhang F,Wang X,Liu Y,Hu C.Isolation,identification and characterization of phytoplankton-lytic bacterium CH-22 against Microcystis aeruginosa[J].Limnology,2011.41(1):70-77
4 Lee J,Cho DH,Ramanan R,Kim BH,Oh HM,Kim HS.Microalgaeassociated bacteria play a key role in the flocculation of Chlorella vulgaris[J].Bioresour Technol,2013,131(2):195-201
5 Chen CY,Yeh KL,Aisyah R,Lee DJ,Chang JS.Cultivation,photobioreactor design and harvesting of microalgae for biodiesel production:a critical review[J].Bioresour Technol,2011,102(1):71-81
6 Powell RJ,Hill RT.Rapid aggregation of biofuel-producing algae by the bacterium Bacillus sp.strain RP1137[J].Appl Environ Microbiol,2013,79(19):6093
7张佳琪,庞宁,杨舒棋,林泓,胡筱敏,姜彬慧.絮凝法采收生物燃料微藻的研究进展[J].环境保护与循环经济,2017,37(3):25-30[Zhang JQ,Pang N,Yang SQ,Lin H,Hu YM,Jiang BH.Advances in research on harvesting biofuel microalgae by flocculation[J].Liaoning Urban Rural Environ Sci Technol,2017,37(3):25-3 0]
8 Christenson L,Sims R.Production and harvesting of microalgae forwastewater treatment,biofuels,and bioproducts[J].Biotechnol Adv2011,29(6):686-702
9 Uduman N,Qi Y,Danquah MK,Forde GM,Hoadley A.Dewatering of microalgal cultures:a major bottleneck to algae-based fuels[J].J Renew Sustain Energ,2010,2(1):23-571
10 Alam MA,Wan C,Guo SL,Zhao XQ,Huang ZY,Yang YL,Chang JS,Bai FW.Characterization of the flocculating agent from the spontaneously flocculating microalga Chlorella vulgaris JSC-7[J].J Biosc Bioeng,2014,1 18(1):29-33
11 McSwain BS,Irvine RL.Composition and distribution of extracellular polymeric substances in aerobic flocs and granular sludge[J].Appl Environ Microbiol,2005,71(2):1051-1057
12 Liu XM,Sheng GP,Luo HW,Zhang F,Yuan SJ,Xu J,Zeng Raymond J;Wu JG,Yu HQ.Contribution of extracellular polymeric substances(EPS)to the sludge aggregation[J].Environ Sci Technol,2010,44(11):4355-4360
13 Sanin FD,Vesilind PA.Effect of centrifugation on the removal of extracellular polymers and physical properties of activated sludge[J].Water Sci Technol,1994,30(8):117-127
14 Yu GH,He PJ,Shao LM.Characteristics of extracellular polymeric substances(EPS)fractions from excess sludges and their effects on bioflocculability[J].Bioresour Technol,2009,100(13):3193-3 198
15 Wilen BM,Jin B,Lant P.The influence of key chemical constituents in activated sludge on surface and flocculating properties[J].Water Res,2003,37(9):2127-2139
16龙向宇,龙腾锐,唐然,李金印,罗太忠.阳离子交换树脂提取活性污泥胞外聚合物的研究[J].中国给水排水,2008,24(3):29-33[Long XY,Long TR,Tang R,Li JY,Luo TZ.Study on extraction of extracellular polymeric substance from activated sludge using cation exchange resin[J].Chin Wa ter Wa stewater,2008,24(3):29-33]
17 Oh H,Lee S,Park M,Kim H.Harvesting of Chlorella vulgaris using a bioflocculant from Paenibacillus sp.AM49[J].Biotechnol Lett,2001,23(15):1229-1234
18 Xu H,Yu G,Jiang H.Investigation on extracellular polymeric substances from mucilaginous cyanobacterial blooms in eutrophic freshwater lakes[J].Chemosphere,2013,93(1):75-81
19 Chen X,Zhang C,Tan L,Wang J.Toxicity of Co nanoparticles on three species of marine microalgae[J].Environ Pollut,2018,236:454-461
20 Chen W,Westerhoff P,Leenheer JA,Booksh K.Fluorescence excitationemission matrix regional integration to quantify spectra for dissolved organic matter[J].Environ Sci Technol,2003,37(24):5701-5710
21 Chabalina LD,Pastor MR,Rico DP.Characterization of soluble and bound EPS obtained from 2 submerged membrane bioreactors by3D-EEM and HPSEC[J].Talanta,2013,115(17):706-712
22 Liu X,Wu S,Zhang D,Shen J,Han W,Sun X,Li J,Wang L.Simultaneous pyridine biodegradation and nitrogen removal in an aerobic granular system[J].J Environ Sci,2018,67(5):318-329
23 Jia F,Yang Q,Liu X,Li X,Li B,Zhang L,Peng,Y.Stratification of extracellular polymeric substances(EPS)for aggregated anammox microorganisms[J].Environ Sci Technol,2017,51(6):3260-3268
24 Hori K,Matsumoto S.Bacterial adhesion:from mechanism to control[J].Biochem Eng J,2010,48(3):424-434
25 Marshall KC,Stout R,Mitchell R.Mechanism of the initial events in thesorption of marine bacteria to surfaces[J].J Gen Microbiol,1971,68(3):337-348
26张兰河,李军,郭静波,贾艳萍,张海丰.EPS对活性污泥絮凝沉降性能与表面性质的影响[J].化工学报,2012,63(6):1865-1871[Zhang L,Li J,Guo J,Jia Y,Zhang H.Effect of EPS on flocculation-sedimentation and surface properties of activated sludge[J].J Chem Ind Eng(Chin).2012,63(6):1865-1871]
27 Kolter R,Greenberg EP.Microbial sciences:the superficial life of microbes[J].Nature,2006,441(7091):300-302
28 Long Z,Quaife B,Salman H,Oltvai ZN.Cell-cell communication enhances bacterial chemotaxis toward external attractants[J].Sci Rep.2017,7(1)
29 Golzarijalal M,Ashtiani FZ,Dabir B.Modeling of microalgal shearinduced flocculation and sedimentation using a coupled CFD-population balance approach[J].Biotechnol Prog,2018,34(1):160-174
30胡沅胜,刘斌,郝晓地,曹亚莉.微藻处理污水中的絮凝分离/采收研究现状与展望[J].环境科学学报,2015,35(1):12-29[Hu YS,Liu B,Hao XD,Cao YL.Current status and outlook of microalgae flocculation in wastewater treatment[J].Acta Sci Circumt,2015,35(1):12-29]
31 Salim S,Bosma R,Vermue MH,Wijffels.Harvesting of microalgae by bio-flocculation[J].J Appl Phycol,2011,23(5):849-855
32 Ozkan A,Berberoglu H.Adhesion of algal cells to surfaces[J].Biofouling,2013,29(4):469-482
33 Zhou D,Zhang C,Fu L,Xu L,Cui X,Li Q,Crittenden J C.Responses of the microalga Chlorophyta sp.to bacterial quorum sensing molecules(N-Acylhomoserine Lactones):aromatic protein-induced self-aggregation[J].Environ Sci Technol,2017,51(6):3490-3498
34 Ding A,Pronk W,Qu FS,Ma J,Li GB,Li K,Liang H.Effect of calcium addition on sludge properties and membrane fouling potential of the membrane-coupled expanded granular sludge bed process[J].J Membr Sci,2015,489:55-63
35 Meng FG,Zhou ZB,Bing-Jie Ni,Zheng X,Huang GC,Jia XS,Li SY,Xiong Y,Kraume M.Characterization of the size-fractionated biomacromolecules:tracking their role and fate in a membrane bioreactor[J].Wa ter Res,2011,45(15):4661-4671
36宿程远,郑鹏,卢宇翔,谢莲,黄纯萍,黄智,陈孟林.海泡石与生物质炭强化厌氧处理养猪废水[J].中国环境科学,2017,37(10):3764-3772[Su CY,Zheng P,Lu YX,Xie L,Huang C, Huang Z,Chen ML.Enhanced efficiency of an anaerobic reactor containing sepiolite or biochar for treatment swine wastewater[J].Chin Environ Sci,2017,37(10):3764-3772]
37王硕,于水利,徐巧,付强,李激.好氧颗粒污泥特性、应用及形成机理研究进展[J].应用与环境生物学报,2014,20(4):732-742[Wang S,Yu SL,Xu Q,Fu Q,Li J.Characteristics,application and formation mechanisms of aerobic granular sludge:recent advances[J].Chin J Appl Environ Biol,2014,20(4):732-742]
38唐堂,王硕,蒋志坚,李激.群感效应与信号分子在污泥颗粒化过程中的作用研究进展[J].应用与环境生物学报,2016,22(4):718-724[Tang T,Wang S,Jiang ZJ,Li J.Progress in study of the function of quorum sensing and cell signaling in the formation of aerobic granular sludge[J].Chin J Appl Environ Biol,2016,22(4):718-724]
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