好氧顆粒污泥的培養(yǎng)過(guò)程

附件2論文中英文摘要作者姓名：倪丙杰論文題目：好氧顆粒污泥的培養(yǎng)過(guò)程、作用機(jī)制及數(shù)學(xué)模擬作者簡(jiǎn)介：倪丙杰，男，1981年11月出生，2006年9月師從于中國(guó)科學(xué)技術(shù)大學(xué)俞漢青教授，于2009年7月獲博士學(xué)位。中文摘要好氧顆粒污泥是活性污泥微生物通過(guò)自固定最終形成的結(jié)構(gòu)緊湊、外形規(guī)則的生物聚集體。它具有相對(duì)密實(shí)的微觀結(jié)構(gòu)、優(yōu)良的沉淀性能、較高濃度的生物體截留和多樣的微生物種群。因此，好氧顆粒污泥技術(shù)作為一種新型的廢水生物處理形式，在城市污水和工業(yè)廢水處理中具有非常廣闊的應(yīng)用前景。好氧顆粒污泥的形成過(guò)程非常復(fù)雜，它的作用機(jī)制涉及到微生物的生長(zhǎng)與競(jìng)爭(zhēng)、氧的傳質(zhì)、底物的擴(kuò)散及微生物產(chǎn)物的形成等各個(gè)方面。本論文系統(tǒng)地研究了好氧顆粒污泥的形成過(guò)程、作用機(jī)制和數(shù)學(xué)模擬，探索了顆粒污泥在好氧和缺氧條件下的胞內(nèi)儲(chǔ)存過(guò)程機(jī)理，深入闡明了顆粒污泥中胞外聚合物和溶解性微生物產(chǎn)物的形成規(guī)律，并首次成功地以低有機(jī)物濃度城市污水為基質(zhì)在中試規(guī)模反應(yīng)器中培養(yǎng)出性能優(yōu)良的好氧顆粒污泥。主要研究?jī)?nèi)容和研究結(jié)果如下：分別采用豆制品加工廢水和混合酸廢水在SBR反應(yīng)器中培養(yǎng)好氧顆粒污泥，基于實(shí)驗(yàn)結(jié)果和形成機(jī)理的分析，實(shí)現(xiàn)了好氧顆粒污泥形成過(guò)程的定量描述。粒污泥在形成過(guò)程中粒徑逐漸增大，沉降速度提高到40mh-1,污泥體積指數(shù)SVI減小至20mLg〔，COD去除效率高于98%；模型能夠很好地定量描述好氧顆粒污泥的形成過(guò)程及基質(zhì)在顆粒內(nèi)部的擴(kuò)散；好氧顆粒污泥的形成過(guò)程可分為適應(yīng)期、快速生長(zhǎng)期和成熟期。通過(guò)好氧顆粒污泥的間歇實(shí)驗(yàn)，探索了顆粒中自養(yǎng)微生物和異養(yǎng)微生物的生長(zhǎng)與競(jìng)爭(zhēng)；根據(jù)實(shí)驗(yàn)結(jié)果修正了ASM3模型，用以描述好氧顆粒SBR反應(yīng)器中自養(yǎng)菌和異養(yǎng)菌的同時(shí)生長(zhǎng)，并分析兩類微生物對(duì)于溶解氧的競(jìng)爭(zhēng)和在顆粒中的空間分布；發(fā)現(xiàn)氨氮和COD在顆粒SBR反應(yīng)器運(yùn)行周期的前1.5小時(shí)內(nèi)分別被自養(yǎng)菌和異養(yǎng)菌消耗完畢，且異養(yǎng)菌消耗更多的溶解氧；自養(yǎng)菌主要位于顆粒的外層，而異養(yǎng)菌則分布于顆粒的外層或者中心。通過(guò)McCarty建立的生物熱能學(xué)方法估算出活性污泥的微生物產(chǎn)率，分析了反應(yīng)器中關(guān)鍵化學(xué)和生物組分的總體計(jì)量學(xué)；與生化反應(yīng)和電子平衡相耦合，建立了分析反應(yīng)器中各種重要化學(xué)組分的動(dòng)態(tài)變化的熱力學(xué)新方法，并同修正后的ASM1模型相結(jié)合，解析了豆制品廢水活性污泥處理系統(tǒng)中C16H24O5N4、CH20、細(xì)胞(C5H7O2N)、H+、NH4+、HCO3「和C02濃度的動(dòng)態(tài)變化規(guī)律。通過(guò)不同初始條件下的顆粒污泥儲(chǔ)存實(shí)驗(yàn)，探索了好氧條件下顆粒污泥的胞內(nèi)儲(chǔ)存機(jī)制和電子轉(zhuǎn)移途徑，實(shí)現(xiàn)了好氧顆粒污泥胞內(nèi)儲(chǔ)存過(guò)程的準(zhǔn)確描述。當(dāng)系統(tǒng)中外源底物過(guò)剩時(shí)，底物被微生物快速利用并形成一定量的儲(chǔ)存物質(zhì)；在外源底物消耗完之后，微生物則會(huì)利用胞內(nèi)儲(chǔ)存物質(zhì)進(jìn)行生長(zhǎng)。采用好氧和缺氧條件下的呼吸速率實(shí)驗(yàn)結(jié)果對(duì)反硝化條件下顆粒污泥的儲(chǔ)存機(jī)理進(jìn)行了探索，發(fā)現(xiàn)其生化機(jī)理過(guò)程主要包括反硝化條件下的水解、缺氧同時(shí)儲(chǔ)存和生長(zhǎng)、缺氧維持以及生物衰減。動(dòng)力學(xué)分析結(jié)果表明，好氧顆粒污泥的反硝化胞內(nèi)儲(chǔ)存過(guò)程N(yùn)UR包括4個(gè)不同的硝酸鹽利用線性階段；根據(jù)缺氧儲(chǔ)存機(jī)理，建立了缺氧校正系數(shù)的估算方法，并解析出好氧顆粒污泥在反硝化條件下的胞內(nèi)儲(chǔ)存過(guò)程。采用凝膠滲透色譜(GPC)和三維熒光光譜(EEM)對(duì)活性污泥胞外多聚物(EPS)的產(chǎn)生過(guò)程進(jìn)行了表征。EPS的形成量及其組分分子量分布依賴于外源底物的利用， EPS量在底物利用過(guò)程中逐漸增加；EPS中的熒光物質(zhì)主成分有兩種:最大激發(fā)發(fā)射波長(zhǎng)位于280/340nm的蛋白類物質(zhì)和最大激發(fā)發(fā)射波長(zhǎng)位于320/400nm的類富里酸物質(zhì)。提出了一套估算活性污泥EPS形成動(dòng)力學(xué)參數(shù)的新方法。在該方法中利用非線性最小二乘法建立目標(biāo)函數(shù)，而采用蒙特卡羅法優(yōu)化目標(biāo)函數(shù)；參數(shù)估計(jì)結(jié)果表明，此方法能夠準(zhǔn)確、簡(jiǎn)便地獲得活性污泥微生物降解有機(jī)物過(guò)程中 EPS的形成動(dòng)力學(xué)參數(shù)值及其相應(yīng)的可靠性信息；外源底物電子的分布途徑為：61%的電子用于細(xì)胞生物量的合成，21%的電子轉(zhuǎn)移到電子受體溶解氧用于生物呼吸產(chǎn)生能量，而剩下的 18%的電子則用于EPS的形成。集成運(yùn)用凝膠滲透色譜、耗氧速率分析、多糖和蛋白測(cè)定、三維熒光光譜、傅立葉變換紅外光譜和溶解性有機(jī)碳測(cè)定等技術(shù)，全面分析了活性污泥SMP的分子量分布、化學(xué)特性及其分類，對(duì)污泥微生物降解底物過(guò)程中SMP的形成及其分類劃分進(jìn)行了定性和定量研究。發(fā)現(xiàn)UAP產(chǎn)生于底物利用階段，其分子量低于290kDa,而B(niǎo)AP為290-5000kDa范圍內(nèi)的生物大分子物質(zhì)，BAP則可進(jìn)一步分為生長(zhǎng)相關(guān)BAP(GBAP)和內(nèi)源相關(guān)BAP(EBAP)。提出了分析SMP、UAP和BAP產(chǎn)生動(dòng)力學(xué)的新方法，并建立了描述SMP形成過(guò)程及其劃分的數(shù)學(xué)模型?；诘孜锢梅e分方程分別建立了UAP和BAP目標(biāo)函數(shù)，運(yùn)用規(guī)劃求解法優(yōu)化目標(biāo)函數(shù)使其最小化，從而得到活性污泥SMP形成的相關(guān)速率系數(shù)、Ks、kBAP以及產(chǎn)率系數(shù)Yh、kuAP的最佳估計(jì)值；在此基礎(chǔ)上描述了3種SMP的產(chǎn)生與底物利用之間的聯(lián)系以及SMP的形成機(jī)理。采用實(shí)驗(yàn)分析和數(shù)學(xué)模擬相結(jié)合的方法闡明了好氧顆粒污泥系統(tǒng)中 EPS、SMP和XSTO的形成和消耗過(guò)程，在將這3種不同微生物代謝產(chǎn)物的電子轉(zhuǎn)移途徑相耦合的基礎(chǔ)上，建立一套統(tǒng)一的EPS、SMP和XSTO的形成動(dòng)力學(xué)模型，分析了反應(yīng)器污泥停留時(shí)間(SRT)對(duì)EPS、SMP和XSTO形成的影響，并提出了基于EPS、SMP和XSTO動(dòng)力學(xué)模型調(diào)控顆粒反應(yīng)器的新方法。對(duì)好氧顆粒污泥自養(yǎng)菌微生物產(chǎn)物的產(chǎn)生規(guī)律、成分特性及數(shù)學(xué)模擬進(jìn)行了系統(tǒng)的研究，闡明了自養(yǎng)菌微生物代謝產(chǎn)物的形成機(jī)理，并對(duì)自養(yǎng)微生物體系中產(chǎn)生的EPS和SMP進(jìn)行了表征；探索了自養(yǎng)微生物體系中異養(yǎng)微生物利用自養(yǎng)微生物代謝產(chǎn)物的生長(zhǎng)規(guī)律；發(fā)現(xiàn)NoB在好氧條件下以亞硝酸鹽作為電子供體進(jìn)行生長(zhǎng)，并產(chǎn)生新的NoB細(xì)胞、UAP、EPS和硝酸鹽；這些自養(yǎng)微生物的代謝產(chǎn)物是自養(yǎng)體系中異養(yǎng)菌細(xì)胞合成的唯一電子供體來(lái)源。首次利用低濃度實(shí)際城市污水在中試規(guī)模反應(yīng)器中培養(yǎng)出性能優(yōu)良的好氧顆粒污泥，探索了低濃度有機(jī)廢水作為底物條件時(shí)污泥顆粒化的關(guān)鍵因素，并且實(shí)現(xiàn)了中試顆粒污泥反應(yīng)器的數(shù)學(xué)模擬。好氧顆粒污泥粒徑適中、沉降性良好、生物活性高；顆粒污泥主要以絲狀菌為骨架，由結(jié)構(gòu)密實(shí)地桿狀細(xì)菌和球菌組成；分析表明體積交換率和沉降時(shí)間是低濃度廢水條件下SBR反應(yīng)器中污泥顆粒化的關(guān)鍵參數(shù)。采用好氧顆粒污泥為接種物，在上流式厭氧污泥反應(yīng)器(UASB)中實(shí)現(xiàn)厭氧氨氧化(ANAMMoX)過(guò)程的快速啟動(dòng)，并對(duì)所形成ANAMMoX顆粒污泥的結(jié)構(gòu)和物化特性以及反應(yīng)器運(yùn)行工況進(jìn)行表征；在此基礎(chǔ)上，采用16SrRNA基因擴(kuò)增和PCR-DGGE等分子生物學(xué)技術(shù)對(duì)啟動(dòng)過(guò)程中顆粒污泥的微生物種群結(jié)構(gòu)變化進(jìn)行定量分析，為拓展好氧顆粒污泥的應(yīng)用領(lǐng)域提供了新的思路。關(guān)鍵詞：好氧顆粒污泥；數(shù)學(xué)模擬；微生物產(chǎn)物；序批式反應(yīng)器(SBR)；中試顆粒污泥反應(yīng)器Formationprocess,functionmechanismandmathematicalmodelingof

theaerobicgranularsludgeNiBing-JieABSTRACTAerobicgranularsludgeisatypeofmicrobialaggregatethroughself-immobilizationandgranulationofthemicroorganismsinactivatedsludge.Aerobicgranuleusuallyhasregularshape,goodsettlingability,compactstructure,highbiomassretention,anddiversityofmicrobialpopulation.Itensuresahighersubstrateremovalefficiency,lessexcesssludgedisposal,lessareaconsumptionandlowercostsforcapitalconstruction,comparedwithactivatedsludgeflocs.Therefore,thisprocesshasbeenregardedasapromisingwastewatertreatmentsystem.However,aerobicgranulationisaverycomplexphenomenon.Therearenumerousinternalinteractionsamongprocessvariables,suchasgrowth,storage,microbialproductsformationandendogenousrespiration,andsludgecharacteristics,includingbiomassdetachment,oxygentransferanddiffusion.Inthisthesis,theformation,thefunctionmechanismandthemathematicalmodelingofaerobicgranularsludgeweresystematicallyexplored.Thestorageprocessesofaerobicgranulesunderbothaerobicandanoxicconditionswereinvestigated.Theproductionofextracellularpolymericsubstances(EPS)andsolublemicrobialproducts(SMP)inaerobicgranularsludgewerealsoexplored.Furthermore,thisworkwasthefirstattempttocultivateaerobicgranulesonlow-strengthmunicipalwastewaterinapilot-scalesequencingbatchreactor(SBR).Maincontentsandresultsareasfollows:AerobicgranulesweresuccessfullycultivatedinSBRsfedwithbothsoybean-processingandfatty-acids-richwastewaters.Basedonexperimentalobservationsandformationmechanismanalysis,theaerobicgranulationprocessintermsofmeanradiusprofileswasquantitativelycharacterized.Inthegranulationprocess,themeandiameterofbioparticlesgraduallyincreased.Theirsettlingvelocityincreasedto40mh-1whiletheirsludgevolumeindex(SVI)decreasedto-i20mLg.Thechemicaloxygendemand(COD)removalefficiencyincreasedto98%.Thedevelopedmodelisapplicabletodescribingtheaerobicsludgegranulationprocessandsubstratediffusionwithingranulesappropriately.Threephasesinthegranuleformationprocesscouldbeclearlydistinguished:initialexponentialgrowthphase,lineargrowthphaseafterwardsandfinalstablephase.Theautotrophicandheterotrophicgrowthandcompetitioninaerobicgranularsludgewereexploredusingabatchexperimentalapproach.TheactivatedsludgemodelNo.3(ASM3)wasmodifiedbasedonexperimentalresultsinordertodescribethesimultaneousautotrophicandheterotrophicgrowthinaerobicgranules.Thedistributionwithingranulesandcompetitionfordissolvedoxygenofautotrophsandheterotrophswereanalyzed.ItwasfoundthatfulloxidationofammoniaandCODbyautotrophsandheterotrophsoccurredwithin1.5h.Theheterotrophsaccountedformajoroxygenconsumptionthantheautotrophs.Theautotrophsweremainlylocatedontheouterlayerofgranules,whereastheheterotrophswerepresentinthecenterofgranules,orontheouterlayerofgranules.WiththebioenergeticmethodologyestablishedbyMcCarty,themicrobialyieldwaspredictedandtheoverallstoichiometricsforbiologicalreactionsinvolvingthekeychemicalandbiologicalspeciesinactivatedsludgewereestablished.ThebioenergeticmethodologywasintegratedwithamodifiedactivatedsludgemodelNo.1(ASM1)toformulateanewapproachtoanalyzetheactivatedsludgeprocess,withthetreatmentofsoybean-processingwastewaterasanexample.ThisapproachwasabletoapproximatelydescribethetreatmentofsoybeanwastewaterbyactivatedsludgeintermsoftheconcentrationdynamicsofC16H24O5N4,CH2O,cell(C5H7O2N),H+,+NH4+,HCO3-andCO2.Theinternalstoragemechanismsandelectronflowsfromtheexternalsubstrateoccurringinaerobicgranulesludgewereexploredwithextensivestorageexperimentsunderdifferentinitialconditions.Thesimultaneousgrowthandstorageprocessesinaerobicgranuleswereaccuratelymodeled.AerobicgranulesinanSBRweresubjectedtoalternativefeastandfamineconditions,andwereabletorapidlytakeupcarbonsubstrateinwastewaterandtostoreitasintracellularstorageproductswhenthesubstratewasinexcesAs.fterthedepletionoftheexternalsubstrate,theaccumulatedstoragepolymerwasutilizedforheterotrophicgrowth.Theinternalstoragemechanismsinaerobicgranulesludgeunderanoxicconditionswereinvestigatedwithrespirometricexperimentalresults.Hydrolysis,simultaneousanoxicstorageandgrowth,anoxicmaintenance,andendogenousdecaywerefoundtobethemainbioreactionprocessesgoverningtheanoxicstorageintheaerobicgranules.Kineticanalysisofnitrateutilizationrate(NUR)indicatesthattheNURofgranules-baseddenitrificationprocessincludedfourlinearphasesofnitratereduction.Furthermore,themethodologyfordeterminingthemostimportantparameterinanoxicstorage,i.e.,anoxicreductionfactor,wasestablishedbasedonananalysisofanoxicstoragemechanisms.Theperformanceofstorageprocessinagranule-baseddenitrificationsystemwasaccuratelymodeled.EPSproducedbymixedmicrobialcommunitywerecharacterizedusinggel-permeatingchromatography(GPC)and3-dimensionalexcitationemissionmatrix(EEM)fluorescencespectroscopymeasurement.TheproductionofEPS,aswellasitsmolecularweight(MW),dependedontheexternalsubstrateutilization.ThequantityofproducedEPSincreasessignificantlyinthesubstrateutilizationprocess.Withtheparallelfactoranalysis(PARAFAC)approach,twocomponentsofthepolymermatrixareidentifiedbytheEEManalysis,oneasproteinsatEx/Em280/340nmandonematrixassociatedasfulvic-acid-likesubstancesat320/400nm.AnovelandconvenientapproachtoevaluatetheEPSproductionkineticswasdeveloped.Theweightedleast-squaresanalysiswasemployedtocalculateapproximatedifferencesinEPSconcentrationbetweenmodelpredictionsandexperimentalresults.AniterativesearchroutineinMonteCarlosimulationwasutilizedforoptimizingtheobjectivefunctionbyminimizingthesumofsquaredweightederrors.ParametersestimationresultsindicatethatthekineticcoefficientsofEPSproductionbyactivatedsludgeandtheirpracticalidentifiabilityinformationcouldbeobtainedaccuratelyandconvenientlywiththisapproach.Electronsfromtheexternalsubstrateweredistributedinthefollowingorder:newbiomasssynthesisof61%,oxygenforrespirationof21%,andEPSof18%.Thesub-fractionsoftheSMP,i.e.,utilization-associatedproducts(UAP)andbiomass-associatedproducts(BAP),excretedbyactivatedsludgewerecharacterizedintermsofformationsequence,MWandchemicalnatures,usingMWanddissolvedorganiccarbon(DOC)measurements,coupledwithoxygenutilizationratedetermination,polysaccharideandproteinmeasurement,3-dimensionalEEMfluorescencespectroscopyandFouriertransforminfraredspectroscopy(FTIR)analysis.TheUAP,producedinthesubstrateutilizationprocess,werefoundtobecarbonaceouscompoundswithanMWlowerthan290kDa.TheBAPweremainlycellularmacromoleculeswithanMWinarangeof290-5000kDa,andwerefurtherclassifiedintothegrowth-associatedBAP(GBAP)andtheendogeny-associatedBAP(EBAP).AnewapproachfordeterminatingSMP,UAPandBAPandtheirproductionkineticswasestablished.Amathematicalmodelwasdevelopedtofurtherquantitativelyandqualitativelydescribetheproductionkineticsandsub-fractionationofSMP.TheobjectivefunctionsofUAPandBAPwereconstructedbasedontheintegratedsubstrateutilizationequation.Aspreadsheetprogramwasutilizedtooptimizetheobjectivefunctionbyminimizingthesumofsquaredweightederrors.Inthisway,thebestestimationsoftheratecoefficients曲,KSandkBAPandthetwoyieldcoefficientsforactivebacteria(YH)andUAP(kUAP)weredetermined.Furthermore,therelationshipsamongtheformationofthethreesub-fractionsoftheSMPandthesubstrateutilization,aswellastheSMPformationmechanisms,wereelucidated.TheformationofEPS,SMPandinternalstorageproducts(XSTO)inaerobicgranularsludgewasinvestigatedusingexperimentalandmodelingapproaches.AnexpandedunifiedmodeldescribingtheproductionandtheconsumptionofEPS,SMP,andXSTOwasformulatedafterintegratingtheelectronflowsfromtheexternalsubstratetoEPS,SMP,andXSTO.Theeffectofthesludgeretentiontime(SRT)ofthereactorontheformationofEPS,SMP,andXSTOintheaerobicgranularsludgewasanalyzed.Thenewmodelcouldbeusedforprocesscontrolandthusfortheoptimizationofaerobic-granule-basedreactors.Theformationmechanism,componentcharacterization,andmathematicalmodelingofthemicrobialproductsofautotrophsintheaerobicgranularsludgewereinvestigatedsystemically.TheformationmechanismofEPSandSMPbyautotrophswasclarified.InformationontheMWandchemicalnaturesofmicrobialproductsinthenitrifyingsludgewasexploredusingEEMfluorescencespectroscopyandGPCmeasurement.Theheterotrophicgrowthonthemicrobialproductsinthenitrifyingsludgefeedingwithnon-organiccarbonsourcewasalsoevaluated.TheaerobicgrowthoftheNOBoccurredattheexpenseofnitriteasanelectrondonorandresultedintheproductionofnewbiomass,UAP,EPS,andnitrate.Theseautotrophicmicrobialproductscouldbeutilizedasthesoleelectronsandcarbonsourcefortheheterotrophicgrowthinautotrophicsystems.Forthefirsttime,aerobicgranuleswithanexcellentsettlingabilitywerecultivatedinapilot-scale