胡小兵,汪 坤,李晶晶,陳紅偉,宋維維,江用彬,2,常 靜,鐘梅英

污泥絮體特征與微型動(dòng)物運(yùn)動(dòng)速度相關(guān)性研究——以自由運(yùn)動(dòng)型微型動(dòng)物為例

胡小兵1,2*,汪 坤1,李晶晶1,陳紅偉1,宋維維1,江用彬1,2,常 靜1,鐘梅英1

(1.安徽工業(yè)大學(xué)建筑工程學(xué)院,安徽 馬鞍山 243002；2.生物膜法水質(zhì)凈化及利用技術(shù)教育部工程研究中心,安徽 馬鞍山 243032)

絮體微觀形態(tài)參數(shù)；PCA法；微型動(dòng)物；運(yùn)動(dòng)速度；活性污泥

活性污泥絮體是由微生物(細(xì)菌、真菌和微型動(dòng)物等)與其代謝產(chǎn)物及所吸附的有機(jī)、無機(jī)物混合組成[1],表面凹凸不平,形態(tài)結(jié)構(gòu)多樣[2].絮體作為活性污泥法中的微型生物處理單元,承擔(dān)著有機(jī)污染物降解的重要作用[3].絮體的結(jié)構(gòu)特征可用于污泥物理特性、生化活性以及廢水處理效果的判斷[4].目前活性污泥絮體的結(jié)構(gòu)特征分析主要有定量圖像分析法[5]和分形理論法[6-7],后者對于絮體結(jié)構(gòu)特征描述主要集中于理論分析與計(jì)算機(jī)數(shù)值模擬分析,缺少對分形結(jié)構(gòu)理論完整性的系統(tǒng)表達(dá)[8].前者主要運(yùn)用顯微成像系統(tǒng)采集絮體圖片,利用定量圖像分析法(QIA)對絮體的平均面積、孔隙面積率、圓度、當(dāng)量直徑、最大Feret直徑等基本參數(shù)進(jìn)行測量[9-10].但分析過多的基本描述參數(shù)將會(huì)加大分析負(fù)擔(dān)與難度,且容易造成基本描述參數(shù)攜帶信息的重復(fù),因此需要運(yùn)用數(shù)據(jù)降維方法高效提取基本描述參數(shù)重要特征,降低分析難度[11].主成分分析(PCA)線性降維由于具有簡單性、可解釋性和高延展性等優(yōu)點(diǎn),使其成為應(yīng)用廣泛的數(shù)據(jù)降維方法[12].

處于較高營養(yǎng)級的微型動(dòng)物(包括原生動(dòng)物和后生動(dòng)物)作為細(xì)菌的主要捕食者,在以活性污泥絮體構(gòu)建的微生態(tài)系統(tǒng)中起著重要作用,其中指示作用是微型動(dòng)物最為重要的功能[13].有研究認(rèn)為溝鐘蟲()、表殼蟲屬(sp.)和累枝蟲屬(sp.)、輪蟲(Rotifer)的出現(xiàn)預(yù)示著出水效果和污泥絮體結(jié)構(gòu)特性良好[14-16].小口鐘蟲()和褶累枝蟲()則會(huì)出現(xiàn)于低MLSS、高SVI的活性污泥絮體環(huán)境中[13,17].另有研究提出累枝蟲屬(sp.)和輪蟲(Rotifer)的大量出現(xiàn)是活性污泥絮體結(jié)構(gòu)惡化,即將解絮膨脹的征兆[18].溝鐘蟲()和褶累枝蟲()對環(huán)境的適應(yīng)能力較強(qiáng),不適宜作為指示生物[19].在絮體結(jié)構(gòu)不太密實(shí)的活性污泥中也出現(xiàn)了累枝蟲屬(sp.)和鐘蟲屬(sp.)[20].同時(shí),通過微型動(dòng)物群落多樣性來指示活性污泥絮體運(yùn)行狀態(tài)[21],又往往存在微型動(dòng)物的鑒別誤判、分類和統(tǒng)計(jì)分析工作量大、耗時(shí)長等問題,限制了其在工程中的實(shí)用性.所以需要采用更為簡便、準(zhǔn)確的方法表征微型動(dòng)物對活性污泥絮體結(jié)構(gòu)特性變化的指示作用.

運(yùn)動(dòng)行為反應(yīng)被視為環(huán)境擾動(dòng)影響的首要指標(biāo)[22-23].當(dāng)環(huán)境條件發(fā)生改變時(shí),微型動(dòng)物在行為學(xué)上的應(yīng)答與反應(yīng)優(yōu)先于物種群落結(jié)構(gòu)變化[24].在污水處理中,活性污泥絮體結(jié)構(gòu)特性變化是影響微型動(dòng)物運(yùn)動(dòng)的一個(gè)重要外部環(huán)境因子.所以采用微型動(dòng)物的運(yùn)動(dòng)方式指示活性污泥絮體結(jié)構(gòu)特性變化,能夠提高分析效果.

本研究使用顯微鏡采集典型微型動(dòng)物的運(yùn)動(dòng)視頻、活性污泥絮體圖片,利用圖像分析軟件進(jìn)行形態(tài)結(jié)構(gòu)特征參數(shù)量化分析,并采用PCA法進(jìn)行活性污泥絮體微觀結(jié)構(gòu)降維處理.探究活性污泥絮體與微型動(dòng)物共存的微生態(tài)系統(tǒng)中,污泥絮體微觀指標(biāo)與微型動(dòng)物運(yùn)動(dòng)參數(shù)的相互影響關(guān)系,為采用典型微型動(dòng)物定量運(yùn)動(dòng)參數(shù)指示活性污泥絮體結(jié)構(gòu)特性提供技術(shù)支持.

1 材料與方法

1.1 試驗(yàn)裝置與用水

本試驗(yàn)采用由透明有機(jī)玻璃制成的高度為90.00cm,外徑為7.50cm,有效容積為3.0L的序批式活性污泥法反應(yīng)器(SBR).反應(yīng)器運(yùn)行周期為12h,包括進(jìn)水5min,曝氣10h,沉淀110min和排水5min.曝氣時(shí)間通過智能控制開關(guān)控制,采用電磁式空氣泵(ACO-005,中國)供氣,通過轉(zhuǎn)子流量計(jì)(LZB-3WB,中國)控制曝氣強(qiáng)度(0.2L/min),維持反應(yīng)器DO在(4.67±0.65)mg/L,反應(yīng)器在室溫(20±5)℃下運(yùn)行.

試驗(yàn)用水取自校園生活區(qū)化糞池,按照化糞池出水與自來水以1:1的比例混合.為了確保進(jìn)水中營養(yǎng)平衡,按照BOD:TN:TP質(zhì)量比為100:5:1的比例在試驗(yàn)用水中添加C6H12O6、NH4Cl和KH2PO4(均為分析純),采用NaHCO3/Na2CO3緩沖劑(pH=9.12)調(diào)節(jié)進(jìn)水pH值.試驗(yàn)用水主要水質(zhì)指標(biāo)如表1所示.

表1 試驗(yàn)進(jìn)水水質(zhì)

1.2 微型動(dòng)物鑒別與運(yùn)動(dòng)參數(shù)分析

在均勻曝氣狀態(tài)下從反應(yīng)器中分時(shí)段取出20mL活性污泥樣品進(jìn)行混合后置于燒杯中,研究的90d中,等間隔的采集了45個(gè)樣品.使用微量移液器(DRAGON,中國)吸出25μL的活性污泥樣品于玻璃載玻片的中央,覆上蓋玻片,置于光學(xué)顯微鏡(OLYMPUS,BX53,日本)下進(jìn)行觀察,參照《微型生物監(jiān)測新技術(shù)》[25]與《環(huán)境微生物圖譜》[26]鑒別到種,屬或是類群.

使用顯微CCD攝像頭(Mshot DC30,中國)與明美成像系統(tǒng)(Mshot Digital Imaging System,中國)進(jìn)行微型動(dòng)物顯微鏡拍攝視頻圖像采集工作,結(jié)合Image J軟件(National Institutes of Health,美國)完成微型動(dòng)物運(yùn)動(dòng)行為參數(shù)的定量表征分析.具體操作和分析方法見文獻(xiàn)[27].

1.3 污泥絮體采集與微觀形態(tài)特征參數(shù)表征

微型動(dòng)物顯微視頻拍攝采集完成后,將顯微鏡調(diào)整至40倍進(jìn)行污泥絮體顯微圖像拍攝,為保證絮體顯微圖像拍攝的完整性,對載玻片所有區(qū)域進(jìn)行逐行不重復(fù)拍攝,每個(gè)載玻片拍攝約60個(gè)圖像,共采集了約2700個(gè)圖像.使用Image-pro Plus 6.0(Media Cybernetics,美國)軟件對所拍攝的圖像進(jìn)行8bit調(diào)整和直方圖均衡的預(yù)處理[28]、自動(dòng)閾值分割[29],中值濾波和開閉運(yùn)算的形態(tài)學(xué)處理[30]、標(biāo)尺轉(zhuǎn)換、殘缺絮體剔除后續(xù)處理操作后,最后導(dǎo)出軟件參數(shù)計(jì)算結(jié)果[31].

研究中共分析16個(gè)絮體微觀形態(tài)參數(shù)[9,32-33].根據(jù)各參數(shù)所表征的物理意義,將其歸屬為絮體大小(SZ)、絮體密實(shí)性(CP)、絮體規(guī)則性(RG)和絮體伸長性(ST)4類特征指標(biāo)[34].對于單個(gè)載玻片所拍攝的60張顯微圖像所提取的16個(gè)微觀形態(tài)特征參數(shù)取平均值作為單次污泥樣品的參數(shù)值.試驗(yàn)中共獲得45組圖像和720組數(shù)據(jù)用于絮體微觀形態(tài)特征參數(shù)相關(guān)性分析.

1.4 污泥絮體微觀形態(tài)參數(shù)PCA分析

為了更加簡潔、高效地表征絮體結(jié)構(gòu),便于分析絮體對微型動(dòng)物運(yùn)動(dòng)的影響,擬采用PCA法在保留大部分信息的情況下進(jìn)一步進(jìn)行降維分析.在考察污泥絮體微觀形態(tài)參數(shù)之間的相互關(guān)系基礎(chǔ)上進(jìn)行PCA,將多項(xiàng)參數(shù)指標(biāo)降維至幾個(gè)綜合參數(shù)指標(biāo)[35].使用SPSS 24(IBM,美國)進(jìn)行主成分分析操作的具體步驟如下:1)進(jìn)行絮體微觀形態(tài)特征參數(shù)的Pearson相關(guān)性分析,計(jì)算相關(guān)系數(shù)矩陣,判斷是否適合進(jìn)行主成分分析;2)對SC和SR指標(biāo)所對應(yīng)的原始參數(shù)矩陣進(jìn)行標(biāo)準(zhǔn)化處理后求出相關(guān)矩陣的特征根和特征向量;3)通過判斷主成分特征值是否大于1確定SC和SR指標(biāo)的主成分個(gè)數(shù),并使得累計(jì)貢獻(xiàn)率達(dá)到75%以上[36];4)得出SC和SR的主成分表達(dá)式,獲得污泥絮體微觀形態(tài)綜合指標(biāo).

采用Pearson相關(guān)性分析污泥絮體微觀形態(tài)綜合指標(biāo)(SC,SR)與自由運(yùn)動(dòng)型微型動(dòng)物運(yùn)動(dòng)速度之間的相關(guān)性特征.

2 結(jié)果與討論

2.1 污泥絮體微觀形態(tài)特征參數(shù)分析

2.1.1 PCA相關(guān)性分析 使用PCA法進(jìn)行絮體微觀形態(tài)特征參數(shù)降維處理的前提是要保證各個(gè)參數(shù)間存在一定的相關(guān)性.

圖1 絮體微觀形態(tài)特征參數(shù)相關(guān)性

此外,FD與Ext間呈顯著負(fù)相關(guān)(=-0.80,< 0.01),說明隨著絮體的分形維數(shù)增加,絮體形狀越規(guī)則,絮體結(jié)構(gòu)緊湊,但絮體充實(shí)度降低[38-39].

表征絮體微觀形態(tài)特征的2類綜合指標(biāo)(SC,SR)不僅內(nèi)部參數(shù)間存在較強(qiáng)相關(guān)性,不同類別參數(shù)間也具有一定的相關(guān)性,約50%的微觀形態(tài)參數(shù)間相關(guān)系數(shù)大于0.3,適合利用PCA法進(jìn)行降維分析,建立絮體大小密實(shí)度(SC)和形狀規(guī)則度(SR)綜合指標(biāo)[40].

2.1.2 基于PCA法的絮體形態(tài)特征指標(biāo)分析 利用PCA法進(jìn)行絮體大小密實(shí)度指標(biāo)(SC)9個(gè)參數(shù)(Amean、Deq、Fmax、、Pconv、、、HR和Ext)的降維分析(表2).

表2 SC參數(shù)的主成分信息提取

注:選取特征值31的成分,下同.

9個(gè)參數(shù)在不同主成分上的因子載荷如圖2所示.

圖2 SC參數(shù)因子載荷圖

式中:Z表示標(biāo)準(zhǔn)化參數(shù)數(shù)據(jù),下同.

故降維后的絮體大小密實(shí)度綜合指標(biāo)

表征絮體形狀規(guī)則度指標(biāo)(SR)的7個(gè)參數(shù)(Asp、ST、RR、BR、FF、RO和FD)經(jīng)過PCA法降維成2個(gè)主成分(表3),累計(jì)保留76.364%信息.

表3 SR參數(shù)的主成分信息提取

7個(gè)參數(shù)在不同主成分上的因子載荷如圖3所示.

圖3 SR參數(shù)因子載荷圖

PC1和PC2表達(dá)式為:

故降維后的絮體形狀規(guī)則度綜合指標(biāo)

2.2 絮體對微型動(dòng)物運(yùn)動(dòng)影響

圖4 銳利楯纖蟲(A. lynceus)運(yùn)動(dòng)參數(shù)

a:絮體間游泳;b:絮體上爬行;c:游泳-爬行互變

2.3 絮體結(jié)構(gòu)與微型動(dòng)物運(yùn)動(dòng)速度的相關(guān)性

圖5 SC和SR與微型動(dòng)物運(yùn)動(dòng)速度(V和W)的相關(guān)系數(shù)

圖6 凹扁前口蟲(F. depressa)運(yùn)動(dòng)軌跡圖

左圖為未受絮體干擾;右圖為受絮體干擾

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Microscopic characteristics of sludge flocs as related to the movement velocity of microfauna: A case study of free-moving microfauna.

HU Xiao-Bing1,2*,WANG Kun1,LI Jing-Jing1,CHEN Hong-Wei1,SONG Wei-Wei1,JIANG Yong-Bin1,2,CHANG Jing1,ZHONG Mei-Ying1

(1.College of Architectural Engineering,Anhui University of Technology,Ma'anshan 243002,China；2.Engineering Research Center of Water Purification and Utilization Technology based on Biofilm Process,Ministry of Education,Ma'anshan 243032,China).,2022,42(8)：3666～3673

microscopic characteristics of flocs；PCA；microfauna；movement velocity；activated sludge

X703

A

1000-6923(2022)08-3666-08

2022-01-05

教育部工程研究中心項(xiàng)目(BWPU2021ZY01,02; BRT19-02);安徽省重點(diǎn)研究與開發(fā)計(jì)劃(202004h07020027)

* 責(zé)任作者,副教授,hxb6608@163.com

胡小兵(1966-),男,安徽涇縣人,副教授,博士,主要從事水處理生物學(xué)與污水生態(tài)處理研究.發(fā)表論文50余篇.