鄭師梅,周啟星,2*,熊紅霞 (.南開大學(xué)環(huán)境科學(xué)與工程學(xué)院,環(huán)境污染過程與基準(zhǔn)教育部重點(diǎn)實(shí)驗(yàn)室,天津市城市生態(tài)環(huán)境修復(fù)與污染防治重點(diǎn)實(shí)驗(yàn)室,天津 30007；2.中國(guó)科學(xué)院沈陽(yáng)應(yīng)用生態(tài)研究所,中國(guó)科學(xué)院陸地生態(tài)過程重點(diǎn)實(shí)驗(yàn)室,遼寧 沈陽(yáng) 006)

腹足綱動(dòng)物毒性測(cè)試候選物種的研究

鄭師梅1,周啟星1,2*,熊紅霞1(1.南開大學(xué)環(huán)境科學(xué)與工程學(xué)院,環(huán)境污染過程與基準(zhǔn)教育部重點(diǎn)實(shí)驗(yàn)室,天津市城市生態(tài)環(huán)境修復(fù)與污染防治重點(diǎn)實(shí)驗(yàn)室,天津 300071；2.中國(guó)科學(xué)院沈陽(yáng)應(yīng)用生態(tài)研究所,中國(guó)科學(xué)院陸地生態(tài)過程重點(diǎn)實(shí)驗(yàn)室,遼寧 沈陽(yáng) 110016)

隨著對(duì)內(nèi)分泌干擾物在腹足綱動(dòng)物中危害性的關(guān)注,軟體動(dòng)物污染生態(tài)問題逐漸受到重視.基于該研究方向現(xiàn)有進(jìn)展,就幾種典型腹足綱動(dòng)物作為化學(xué)品測(cè)試受試生物的候選物種進(jìn)行了論述,認(rèn)為4種前鰓亞綱生物(新西蘭泥蝸、魚盤螺、福壽螺和銅銹環(huán)棱螺)和1種肺螺亞綱生物(靜水椎實(shí)螺)具有生態(tài)意義,可以作為標(biāo)準(zhǔn)受試生物來(lái)發(fā)展.相對(duì)于其他國(guó)外學(xué)者提議的 4種生物,我國(guó)學(xué)者提議的銅銹環(huán)棱螺具有本土意義,在化學(xué)品毒性測(cè)試過程中應(yīng)該受到重視.在以上論述基礎(chǔ)上,對(duì)腹足綱動(dòng)物毒性測(cè)試今后研究的發(fā)展方向進(jìn)行了展望.

化學(xué)品；受試生物；腹足綱動(dòng)物；污染生態(tài)問題；生態(tài)意義

在化學(xué)品毒性測(cè)試中,受試生物的選擇對(duì)正確評(píng)價(jià)化學(xué)品影響很大.自20世紀(jì)80年代以來(lái),經(jīng)濟(jì)合作發(fā)展組織(OECD)、國(guó)際標(biāo)準(zhǔn)化組織(ISO)等國(guó)際組織致力于將化學(xué)品測(cè)試以及基準(zhǔn)研究的受試生物動(dòng)物化.OECD推薦的受試生物有以下幾類:魚類包括斑馬魚(Brachydanio rerio)、黑頭軟口鰷(Pimephales promelas)、鯉魚(Cyprinus carpio)、日本青鳉(Oryzias latipes)、藍(lán)鰓太陽(yáng)魚(Lipomis macrochirus)、孔雀魚(Poecilia reticulata)和虹鱒(Oncorhynchus mykiss)[1];沉積物測(cè)試生物是搖蚊幼蟲(Chironomus tentans)[2]. ISO推薦斑馬魚、藍(lán)鰓太陽(yáng)魚、黑頭軟口鰷和孔雀魚作為淡水魚的代表[3].美國(guó)環(huán)境保護(hù)局(USEPA)推薦搖蚊幼蟲、端足類淡水蝦(Hyalella azteca)和夾雜帶絲蚓(Lumbriculus varigatus)作為沉積物的受試生物[4].美國(guó)試驗(yàn)與材料協(xié)會(huì)(ASTM)推薦的沉積物受試生物除了包括 USEPA推薦的生物外,還包括蜉蝣幼蟲(Hexagenia limbata)、大型蚤(Daphnia magna)、網(wǎng)紋水蚤(Ceriodaphnia dubia)和顫蚓(Tubifex tubifex)等[5].而我國(guó)近幾年開發(fā)的受試生物主要是魚類,包括劍尾魚(Xiphophorus helleri)、稀有鮈鯽(Gobiocypris rarus, 我國(guó)特有種)和紅鯽(Carassius auratus)等[3,6].可見,目前發(fā)展的實(shí)驗(yàn)生物主要是脊椎動(dòng)物中的魚類和無(wú)脊椎動(dòng)物中的節(jié)肢動(dòng)物,卻缺少了無(wú)脊椎動(dòng)物很重要的一部分——軟體動(dòng)物,特別是其中的腹足綱動(dòng)物.雖然USEPA在1985年提出水質(zhì)基準(zhǔn)推導(dǎo)準(zhǔn)則時(shí),將軟體動(dòng)物納入到了受試物種體系中,卻不是如脊椎動(dòng)物和節(jié)肢動(dòng)物一樣的必選物種[7],這說明推導(dǎo)的水質(zhì)基準(zhǔn)難以為軟體動(dòng)物提供應(yīng)有的保護(hù)作用.1995年USEPA發(fā)展了一個(gè)基于腹足綱前鰓亞綱生物紅鮑螺(Haliotis rufescens)幼體發(fā)育的方法用來(lái)檢測(cè)廢水的毒性[8],但不是針對(duì)化學(xué)品的測(cè)試.

1 腹足綱動(dòng)物毒性測(cè)試的生態(tài)意義

軟體動(dòng)物超過了130000種,共有7個(gè)綱,其中已知的 110000種腹足綱動(dòng)物占軟體動(dòng)物的85%[9],其物種的重要生態(tài)意義不應(yīng)該被忽略.蝸牛是腹足綱物種的重要代表,雖然已經(jīng)被用作湖泊營(yíng)養(yǎng)級(jí)水平的指示生物[10]和污染的指示生物

[11-12],但是這只是腹足綱物種生態(tài)意義的部分體現(xiàn),其生態(tài)意義的重要性還沒有被完全的認(rèn)識(shí)和開發(fā).

資料表明,動(dòng)物對(duì)化學(xué)品的敏感性存在差異,不論是脊椎動(dòng)物與無(wú)脊椎動(dòng)物之間,還是脊椎動(dòng)物之間、無(wú)脊椎動(dòng)物之間.Oetken等[13]研究發(fā)現(xiàn),抗癲癇藥卡巴咪嗪(carbamazepine)對(duì)夾雜帶絲蚓和新西蘭泥蝸(Potamopyrgus antipodarum)沒有顯著影響,而對(duì)搖蚊幼蟲有明顯的劑量-效應(yīng)關(guān)系.Nassef等[14]研究發(fā)現(xiàn),卡巴咪嗪對(duì)日本青鳉的進(jìn)食行為和游泳速度都產(chǎn)生影響.而 Nentwig等[15]發(fā)現(xiàn),抗抑郁藥氟西汀(fluoxetine)對(duì)搖蚊幼蟲沒有明顯的劑量-效應(yīng)影響,卻使夾雜帶絲蚓的生殖有略微增加,并且造成了新西蘭泥蝸的生殖率大幅度降低.Foran等[16]發(fā)現(xiàn),氟西汀對(duì)日本青鳉的產(chǎn)卵率、受精率或者受精卵的孵化沒有明顯影響.Stuart等[17]發(fā)現(xiàn),肥鰓螺(Gillia altilis)對(duì)五氯苯酚的敏感性比金魚和黑頭呆魚的高,而Robertson等[18]卻發(fā)現(xiàn)殺蟲劑二嗪農(nóng)(diazinon)對(duì)肥鰓螺的LC50比對(duì)藍(lán)鰓太陽(yáng)魚,虹鱒魚和藍(lán)點(diǎn)石斑的大.由以上結(jié)果可知,用單一物種來(lái)鑒定所有的環(huán)境污染問題是存在矛盾的,基于魚類和節(jié)肢動(dòng)物的化學(xué)品測(cè)試體系的代表性有待完善,需要把軟體動(dòng)物特別是腹足綱動(dòng)物這部分補(bǔ)充進(jìn)來(lái).

實(shí)地檢測(cè)數(shù)據(jù)表明,軟體動(dòng)物對(duì)污染物有很高的敏感性,并對(duì)污染物有指示作用和富集作用[19-28],其中最經(jīng)典的例子是有機(jī)錫導(dǎo)致至少150種海洋前鰓亞綱動(dòng)物的性畸變[9,24-28],一些敏感的新腹足目種類在三丁基錫污染嚴(yán)重的地區(qū)已經(jīng)滅絕[24],而脊椎動(dòng)物或節(jié)肢動(dòng)物未受到或受到很小的影響.因此用軟體動(dòng)物作為毒性試驗(yàn)生物,能填補(bǔ)軟體動(dòng)物缺失的生態(tài)意義.

2 腹足綱動(dòng)物毒性測(cè)試最重要的候選物種

2.1 新西蘭泥蝸

新西蘭泥蝸屬于腹足綱(Gastropoda)的前鰓亞綱(Prosobranchia)角崔螺科(Hydrobiidae).它原產(chǎn)于新西蘭,后來(lái)被引入到世界其他地方,在小溪、河流和湖泊等淡水生態(tài)系統(tǒng)中分布廣泛,在河口等咸水中也有分布.雌雄異體,有單性生殖和雙親生殖兩種,生殖方式為卵胎生.成螺的平均殼高在4.3mm左右,最高能夠達(dá)到6mm.它們從植物、石頭或者沉積物的表面刮食碎屑、藻類和細(xì)菌[29].

Duft等[29]提出3種前鰓亞綱生物作為評(píng)價(jià)內(nèi)分泌干擾物的候選物種,分別是新西蘭泥蝸、大 羊 角 螺 (Marisa cornuarietis,瓶 螺 科Ampullariidae)和網(wǎng)紋織螺(Nassarius reticulatus,蛾螺科Buccinidae).這3種生物對(duì)內(nèi)分泌干擾物都有很高的敏感性,見表 1,但是新西蘭泥蝸在實(shí)驗(yàn)操作方面的優(yōu)勢(shì)最多,所以Duft等最終選擇該生物作為候選物種,并推薦該生物28d的繁殖實(shí)驗(yàn)作為標(biāo)準(zhǔn)測(cè)試.新西蘭泥蝸的優(yōu)勢(shì)依據(jù)為新西蘭泥蝸和大羊角螺已經(jīng)在實(shí)驗(yàn)室成功培養(yǎng),具有實(shí)際應(yīng)用的優(yōu)勢(shì);歐洲的新西蘭泥蝸只有雌性螺,進(jìn)行單性生殖,而來(lái)自單性的個(gè)體對(duì)脅迫因子反應(yīng)的差異性比來(lái)自雙性種類的少;由表1看到,新西蘭泥蝸的測(cè)試終點(diǎn)容易處理,實(shí)驗(yàn)周期相對(duì)較短.而大羊角螺和網(wǎng)紋織螺的測(cè)試需要先進(jìn)的技術(shù)檢測(cè)性畸性,且實(shí)驗(yàn)周期相對(duì)較長(zhǎng).

表1 新西蘭泥蝸、大羊角螺和網(wǎng)紋織螺的實(shí)驗(yàn)參數(shù)[29]Table 1 The experimental parameters of Potamopyrgus antipodarum, Marisa cornuarietis and Nassarius reticulates[29]

2.2 靜水椎實(shí)螺

靜水椎實(shí)螺(Lymnaea stagnalis)是腹足綱肺螺亞綱(Pulmonata)的椎實(shí)螺科(Lymnaeidae)物種,體型為中大型,殼高可達(dá)60mm以上,棲息于緩流或靜水水域,食草性,雌雄同體,主要是自體受精,也存在異體受精[30].

Lagadic等[30]指出,從1960開始,不斷有報(bào)道稱肺螺暴露在大量異型物質(zhì)(包括在無(wú)脊椎動(dòng)物或者脊椎動(dòng)物中已知的內(nèi)分泌干擾物)下,其繁殖行為會(huì)發(fā)生改變.內(nèi)分泌干擾物不僅直接與受體結(jié)合,也間接通過對(duì)荷爾蒙的生產(chǎn)、可得性或者代謝相關(guān)的生物化學(xué)過程的干擾調(diào)節(jié)內(nèi)源荷爾蒙水平,或者通過調(diào)節(jié)受體來(lái)干擾[30-31].所以,肺螺很可能正在遭受與前鰓亞綱螺的性畸形不同的內(nèi)分泌干擾危害,然而此危害未受到重視.

同時(shí),EDIETA (Endocrine Disruption in Invertebrates: Endocrinology, Testing and Assessment)工作組中的deFur等在1999年的報(bào)道中指出,在非前鰓亞綱螺——靜水椎實(shí)螺和紫貽貝(Mytilus edulis)中可能發(fā)生了非有機(jī)錫內(nèi)分泌干擾物的影響[32],具體指,DDT和(4-氯-2-甲基苯氧基)乙酸甲酯導(dǎo)致靜水椎實(shí)螺的產(chǎn)卵率下降[33],金屬鎘明顯抑制紫貽貝雌性和雄性性腺卵泡的發(fā)育,同時(shí)對(duì)產(chǎn)卵頻率有刺激作用[34].由于靜水椎實(shí)螺的神經(jīng)荷爾蒙控制的繁殖已經(jīng)完全被研究清楚,幾種荷爾蒙多肽的結(jié)構(gòu)和受體的主要結(jié)構(gòu)已經(jīng)建立,所以Lagadic等[30]提議將靜水椎實(shí)螺作為內(nèi)分泌干擾物對(duì)肺螺影響的模式生物.

2.3 魚盤螺

魚盤螺(Valvata piscinalis)屬于腹足綱前鰓亞綱的盤螺科(Valvatidae),雌雄同體,異體受精,濾食或者刮食小顆粒的營(yíng)養(yǎng)碎屑、硅藻和細(xì)菌.通常存在于松軟沉積物和泥漿的表層,在運(yùn)河、水溝、池塘、湖泊以及河流等的水生態(tài)系統(tǒng)都能發(fā)現(xiàn)它,在歐洲、加拿大、美國(guó)和亞洲西部,魚盤螺通常是優(yōu)勢(shì)種[35].

表2 魚盤螺的生命周期參數(shù)[35]Table 2 Life cycle parameters for Valvata piscinalis[35]

表3 魚盤螺實(shí)驗(yàn)的推薦[35]Table 3 Recommendations for testing with Valvata piscinalis [35]

Ducrot等[35]將魚盤螺作為沉積物毒性測(cè)試的一種新的候選生物,并提出了培養(yǎng)和測(cè)試的實(shí)驗(yàn)方案.該螺的生物/生態(tài)特點(diǎn)跟其他的腹足綱生物相似,其對(duì)化合物的敏感性具有代表性.該生物在實(shí)驗(yàn)室里被成功培養(yǎng)了六代,由表2看到,實(shí)驗(yàn)室中的新生螺和幼體的生長(zhǎng)速率以及產(chǎn)卵量都比實(shí)地條件的高.Ducrot等推薦,該螺從胚胎到成螺都可以作為毒性測(cè)試生物,見表3.

2.4 福壽螺

福壽螺(Pomacea lineata)是腹足綱前鰓亞綱的瓶螺科生物,是一種兩棲類腹足動(dòng)物,能離開水體短暫生活.個(gè)體大,能達(dá)到70mm左右,為雌雄異體,體內(nèi)受精,體外發(fā)育的卵生動(dòng)物.多集群棲息于池邊淺水區(qū),是以植物性餌料為主的雜食性螺類[36].

Melo等[36]將福壽螺作為巴西東北部本土動(dòng)植物受試生物的代表,利用篩選法半定量地確定不依賴時(shí)間的 LC50和依賴時(shí)間 ILC50.用新生的(4d)、饑餓的福壽螺作為實(shí)驗(yàn)生物,如果第4d或者5d內(nèi)急性毒性明顯(即死亡明顯),研究者應(yīng)繼續(xù)實(shí)驗(yàn)直至實(shí)驗(yàn)生物的死亡停止或者對(duì)照的存活率低于90%.延長(zhǎng)的實(shí)驗(yàn)期間(一般10～14d)能夠看到實(shí)驗(yàn)濃度之間更加清晰的區(qū)別,一般足夠分辨不依賴時(shí)間的最低觀察死亡濃度(LOMC)和無(wú)觀察死亡濃度(NOMC) (分別對(duì)應(yīng)于最低觀察效應(yīng)濃度LOEC,無(wú)觀察效應(yīng)濃度NOEC).

2.5 銅銹環(huán)棱螺

銅銹環(huán)棱螺(Bellamya aeruginosa)屬于腹足綱前鰓亞綱的田螺科(Viviparidae),個(gè)體中等大小,能達(dá)到27mm,雌雄異體,卵胎生.在我國(guó)河流、湖泊、水庫(kù)、水溝等淡水水體中分布廣,數(shù)量大,主要棲居在富含有機(jī)物的沉積物中,以攝食沉積物中有機(jī)碎屑、細(xì)菌和藻類為生.該物種還是一些魚類(如青魚)的食物,同時(shí)也被人們食用[37].

Ma等[37]研究了銅銹環(huán)棱螺在實(shí)驗(yàn)室條件下的生命特征,建立了該物種的實(shí)驗(yàn)室培養(yǎng)和沉積物毒性評(píng)價(jià)的實(shí)驗(yàn)方案,推薦該生物作為沉積物測(cè)試的候選物種.與卵生腹足動(dòng)物(如魚盤螺)相比,銅銹環(huán)棱螺的卵胎生的生長(zhǎng)特點(diǎn)有助于培養(yǎng),因?yàn)樵谄渖缙陔A段不需要特別的照顧.同時(shí)與魚盤螺和新西蘭泥蝸相比,Ma等認(rèn)為銅銹環(huán)棱螺有明顯的優(yōu)勢(shì),主要體現(xiàn)在它的個(gè)體相對(duì)較大(成螺比另外兩種高出 17mm),有助于實(shí)驗(yàn)的處理.基于銅銹環(huán)棱螺的生活史,Ma等推薦幼螺(7d～28d)、中螺(28～112d)和成螺(＞ 112d)都可以用來(lái)評(píng)價(jià)沉積物毒性.

3 存在問題

Giesy等[38]在1989年選擇沉積物毒性測(cè)試的受試生物時(shí),雖然他們認(rèn)為螺類和蛤類生物對(duì)研究很有用,但是他們也指出了該類生物作為受試生物的局限性:很難準(zhǔn)確地確定終點(diǎn)(一般指死亡);確定實(shí)際的暴露劑量存在困難;培養(yǎng)該類生物存在困難.基于以上考慮,以往未將該類生物列入受試物種.

雖然現(xiàn)代方法學(xué)的發(fā)展解決了以上的局限性[35],但是到目前為止,軟體動(dòng)物還沒有常規(guī)的用于化學(xué)品毒性分析中.在 USEPA2005年的生態(tài)毒理受試物種數(shù)據(jù)庫(kù)中僅有47種軟體動(dòng)物被用于實(shí)驗(yàn)室的毒理測(cè)試[35],隨著近幾年對(duì)該類物種生態(tài)意義關(guān)注度的增加,到2010年6月數(shù)據(jù)庫(kù)中蝸牛的種類增加到180種[39].盡管種類豐富度增加,同時(shí)有大量關(guān)于軟體動(dòng)物毒理實(shí)驗(yàn)的文獻(xiàn),但是到目前仍未發(fā)展一種淡水的或者海水的軟體動(dòng)物作為化學(xué)品測(cè)試的標(biāo)準(zhǔn)受試生物.

對(duì)于新西蘭泥蝸,Duft等[29]指出,盡管該生物對(duì)雌性和雄性激素均有高敏感性,但是具有一般生殖毒性的其他物質(zhì)(如重金屬和多環(huán)芳烴)也能產(chǎn)生與內(nèi)分泌干擾物相似的影響,因此研究者要排除一般生殖毒性物質(zhì)的影響.實(shí)際上,新西蘭泥蝸 28d的繁殖測(cè)試,是 EDIETA工作組為OECD發(fā)展無(wú)脊椎動(dòng)物內(nèi)分泌測(cè)試推薦的實(shí)驗(yàn)方案.對(duì)于靜水椎實(shí)螺,Lagadic等[30]指出,盡管充分了解其神經(jīng)荷爾蒙控制繁殖的內(nèi)分泌系統(tǒng),但是在現(xiàn)有內(nèi)分泌學(xué)的知識(shí)水平上還不能清楚的檢測(cè)化學(xué)品誘導(dǎo)的內(nèi)分泌干擾,因此針對(duì)內(nèi)分泌干擾物對(duì)靜水椎實(shí)螺影響的大部分研究,都是從整體方法中間接的探索可能的根源,而且涉及到的機(jī)制是否為內(nèi)分泌干擾仍不清楚.

對(duì)于內(nèi)分泌干擾物的測(cè)試,Matthiessen等[24]指出,目前的研究表明,軟體動(dòng)物特別是蝸牛是對(duì)內(nèi)分泌干擾物最敏感的生物.deFur等在內(nèi)分泌干擾物對(duì)無(wú)脊椎動(dòng)物的影響中指出[32],在發(fā)展內(nèi)分泌干擾物的測(cè)試中,昆蟲和前鰓亞綱蝸牛是最有希望的候選物種. Matthiessen[40]認(rèn)為新西蘭泥蝸暴露 28d不足以覆蓋所有可能的內(nèi)分泌過程,因此提議將軟體動(dòng)物的整個(gè)生命周期作為實(shí)驗(yàn)周期.考慮到靜水椎實(shí)螺的生命周期比新西蘭泥蝸的短,故Matthiessen[40]建議靜水椎實(shí)螺作為整個(gè)生命周期的實(shí)驗(yàn)生物,新西蘭泥蝸?zhàn)鳛椴糠稚芷诘膶?shí)驗(yàn)生物.同樣,Segner等[41]考慮到對(duì)無(wú)脊椎動(dòng)物內(nèi)分泌系統(tǒng)的有限了解,也認(rèn)為無(wú)脊椎動(dòng)物的內(nèi)分泌干擾物實(shí)驗(yàn)更適于在整個(gè)生命周期測(cè)試,并認(rèn)為整個(gè)生命周期測(cè)試是評(píng)價(jià)無(wú)脊椎動(dòng)物內(nèi)分泌干擾物的黃金標(biāo)準(zhǔn).

對(duì)于魚盤螺,Ducrot等[35]比較了加標(biāo) Zn沉積物對(duì)魚盤螺和搖蚊幼蟲的生長(zhǎng)毒性.魚盤螺的LOEC為624mg/kg,而搖蚊幼蟲第2代、第3代和第4代的LOEC分別是953、1382、1400mg/kg,都高于魚盤螺.同時(shí),該螺水體中Zn的24h LC50為31.6mg/L也低于文獻(xiàn)[42]中大型蚤的35.4mg/L.這些結(jié)果表明魚盤螺對(duì)Zn的短期測(cè)試和長(zhǎng)期測(cè)試的敏感性均比兩種標(biāo)準(zhǔn)實(shí)驗(yàn)物種高.

對(duì)于福壽螺,Melo等[36]指出,該螺在毒性研究中不具有代表性,但是當(dāng)它饑餓的時(shí)候,對(duì)低濃度水平的工業(yè)污水和除草劑反應(yīng)的敏感性和一致性在可接受的信用區(qū)間.新生螺對(duì)百草枯(paraquat)的 ILC50和 LC50分別是 0.15和0.35mg/L,LOMC和 NOMC分別是 0.12和0.07mg/L,與文獻(xiàn)[43]中的水蚤、鉤蝦、石蠅、鏢水蚤和波魚相比,福壽螺的LC50要低一個(gè)數(shù)量級(jí).此實(shí)驗(yàn)方案獨(dú)特之處在于饑餓過程,實(shí)驗(yàn)動(dòng)物的敏感性因此增加.而Coller等[44]利用Melo的方法研究除草劑農(nóng)達(dá)(Round-up)的毒性時(shí)發(fā)現(xiàn),不能清晰的分辯死亡是由除草劑的毒性導(dǎo)致的還是饑餓導(dǎo)致的,所以Coller將饑餓過程去掉,喂食新生螺,選擇體重增長(zhǎng)率替代死亡作為反應(yīng)參數(shù),并證明這個(gè)參數(shù)更好.Melo實(shí)驗(yàn)中得到的百草枯的LC50使 Coller實(shí)驗(yàn)中的生物在 6d內(nèi)全部死亡,說明在存活率基礎(chǔ)上估算出來(lái)的LC50將生物放在了一個(gè)極其脆弱的生存水平上. Coller還發(fā)現(xiàn),延長(zhǎng)高濃度農(nóng)達(dá)的暴露時(shí)間,以死亡率為依據(jù),螺未顯示出更高的敏感性.

關(guān)于銅銹環(huán)棱螺,加標(biāo) Cu沉積物對(duì)幼螺的存活率和生長(zhǎng)的 LOEC為 195μg/g,低于成螺的570 μg/g,表明幼體對(duì)加標(biāo)Cu沉積物的敏感性比成螺高[37].幼螺和成螺的LOEC比文獻(xiàn)[45]中端足類淡水蝦和搖蚊幼蟲的(＞2010μg/g)都要低,說明銅銹環(huán)棱螺比兩個(gè)標(biāo)準(zhǔn)受試生物對(duì)沉積物的Cu更為敏感.

對(duì)于選擇實(shí)驗(yàn)最適生物,Buikema等[46]、Giesy等[38]以及 ASTM[47]提出了相似的標(biāo)準(zhǔn):生物必須代表一種對(duì)于分類、營(yíng)養(yǎng)級(jí)或者生態(tài)位很重要的生態(tài)組;生物必須在環(huán)境中廣泛存在,全年可得,且在實(shí)驗(yàn)室容易培養(yǎng),遺傳穩(wěn)定;研究者應(yīng)該知道其生理學(xué)、遺傳學(xué)、分類學(xué)、行為學(xué)等方面的信息;生物對(duì)毒物的反應(yīng)必須是敏感的、一致的、可以檢測(cè)的;生物個(gè)體必須對(duì)疾病和身體傷害有抵抗.以上標(biāo)準(zhǔn)對(duì)候選生物來(lái)說,最重要的方面是對(duì)污染物的敏感性.因此,除了新西蘭泥蝸和靜水椎實(shí)螺,對(duì)于其余3種候選生物來(lái)說,還欠缺大量污染物的急性和慢性的敏感性信息.

文獻(xiàn)中推薦的5種腹足綱候選生物有4種前鰓亞綱生物和1種肺螺亞綱生物.與前鰓亞綱螺不同,肺螺亞綱生物均無(wú)厴,對(duì)毒物不能回避,所以對(duì)毒物比較敏感,因此在發(fā)展腹足綱模式受試生物的時(shí)候應(yīng)該適當(dāng)增加該類生物的比重.該類生物用肺呼吸,適合于水體毒性實(shí)驗(yàn)而不適合沉積物毒性實(shí)驗(yàn)[37].

4 對(duì)發(fā)展我國(guó)模式生物毒性測(cè)試的一點(diǎn)思考

在建立化學(xué)品測(cè)試方面,我國(guó)落后于美國(guó)、歐洲等發(fā)達(dá)國(guó)家和地區(qū).我國(guó)化學(xué)品環(huán)境管理的技術(shù)導(dǎo)則主要是參考OECD的相關(guān)框架和內(nèi)容而制定,如“化學(xué)品測(cè)試導(dǎo)則(HJ/T 153-2004)”、“新化學(xué)品危害評(píng)估導(dǎo)則(HJ/T 154-2004)”和“化學(xué)品測(cè)試合格實(shí)驗(yàn)室導(dǎo)則(HJ/T 155-2004)”.《水和廢水檢測(cè)分析方法》[48]和《化學(xué)品測(cè)試方法》[49]推薦的實(shí)驗(yàn)生物除了我國(guó)的稀有鮈鯽和劍尾魚外,其余都是國(guó)際通用物種.由于生態(tài)系統(tǒng)的敏感種和優(yōu)勢(shì)種具有地域特性,因此使用國(guó)際通用物種不能真實(shí)的反映化學(xué)品對(duì)我國(guó)本土生物的影響,勢(shì)必會(huì)對(duì)我國(guó)本土生物的保護(hù)造成欠缺,甚至可能產(chǎn)生不可恢復(fù)的危害.因此發(fā)展適合我國(guó)本土的實(shí)驗(yàn)生物對(duì)于保護(hù)我國(guó)的自然環(huán)境和生態(tài)系統(tǒng)很重要.目前軟體動(dòng)物的標(biāo)準(zhǔn)受試生物在國(guó)際上還是個(gè)技術(shù)空白,國(guó)際組織(如OECD)和某些國(guó)家正在發(fā)展軟體動(dòng)物實(shí)驗(yàn)準(zhǔn)則,將軟體動(dòng)物列入化學(xué)品測(cè)試準(zhǔn)則的受試生物中.我國(guó)學(xué)者也應(yīng)該認(rèn)識(shí)到軟體動(dòng)物長(zhǎng)期處于欠保護(hù)的事實(shí),發(fā)展符合我國(guó)生態(tài)系統(tǒng)的軟體動(dòng)物尤其是腹足綱動(dòng)物的化學(xué)品測(cè)試準(zhǔn)則.

我國(guó)腹足綱動(dòng)物毒性測(cè)試的發(fā)展水平低于發(fā)達(dá)國(guó)家,目前可供選擇用來(lái)發(fā)展模式生物的實(shí)驗(yàn)物種有限.淡水腹足動(dòng)物的研究主要集中在田螺科和椎實(shí)螺科,海洋腹足動(dòng)物的研究相對(duì)較多,其中很多的研究是針對(duì)有機(jī)錫內(nèi)分泌干擾的毒性.在這些研究中,只有一種淡水腹足綱生物作為候選物種發(fā)展,那就是 Ma等[37]提議的銅銹環(huán)棱螺,作為我國(guó)沉積物實(shí)驗(yàn)的候選物種.國(guó)外學(xué)者提議的新西蘭泥蝸在亞洲從未被發(fā)現(xiàn),因此發(fā)展銅銹環(huán)棱螺對(duì)我國(guó)具有本土意義.劉保元[50]研究過水體污染物對(duì)銅銹環(huán)棱螺的影響,所以銅銹環(huán)棱螺還可以作為水體毒性的實(shí)驗(yàn)生物來(lái)發(fā)展.

5 今后研究展望

5.1 開發(fā)我國(guó)本土腹足綱受試生物

鑒于我國(guó)腹足綱毒性測(cè)試發(fā)展落后的情況,我國(guó)科研工作者應(yīng)該開發(fā)更多具有本土意義的物種.除了前鰓亞綱螺外,肺螺在我國(guó)分布也很廣,因此也應(yīng)該考慮將肺螺發(fā)展為受試生物.由于內(nèi)分泌干擾物對(duì)生物危害的嚴(yán)峻性,我國(guó)學(xué)者應(yīng)該結(jié)合和借鑒國(guó)際內(nèi)分泌干擾物測(cè)試的發(fā)展,開發(fā)我國(guó)本土的腹足綱內(nèi)分泌干擾物實(shí)驗(yàn)生物.

目前我國(guó)正在發(fā)展我國(guó)生態(tài)系統(tǒng)的水質(zhì)基準(zhǔn)[51],而面臨的最大困難是缺少本土的模式受試物種[52],因此開發(fā)我國(guó)本土的腹足綱測(cè)試物種將為水質(zhì)基準(zhǔn)的制定提供技術(shù)支持,同時(shí)避免了USEPA在制定水質(zhì)基準(zhǔn)時(shí)對(duì)軟體動(dòng)物不重視的問題.

5.2 優(yōu)化新西蘭泥蝸和靜水椎實(shí)螺的實(shí)驗(yàn)

新西蘭泥蝸和靜水椎實(shí)螺的實(shí)驗(yàn)程序在形成國(guó)際化準(zhǔn)則之前,需要大量的實(shí)驗(yàn)優(yōu)化和有效化,建立一個(gè)具有充分說服力和最小成本的實(shí)驗(yàn)準(zhǔn)則[40].這需要生物對(duì)更多內(nèi)分泌干擾物的敏感性信息,最佳實(shí)驗(yàn)時(shí)間(28d或者56d),還需要實(shí)驗(yàn)新西蘭泥蝸?zhàn)鳛樗w毒性生物的情況,以及兩種螺繁殖模式對(duì)軟體動(dòng)物整體的代表性程度,更深層次的需要調(diào)查種群反應(yīng)的基因變化.

5.3 重視無(wú)脊椎動(dòng)物內(nèi)分泌系統(tǒng)的研究

由于對(duì)無(wú)脊椎動(dòng)物內(nèi)分泌系統(tǒng)知識(shí)的匱乏,當(dāng)前內(nèi)分泌干擾物測(cè)試的實(shí)驗(yàn)生物主要是魚類和兩棲動(dòng)物[53-54],那么無(wú)脊椎動(dòng)物能否在內(nèi)分泌干擾物測(cè)試中作為實(shí)驗(yàn)生物, 起決定性作用的無(wú)疑是無(wú)脊椎動(dòng)物內(nèi)分泌系統(tǒng)的研究.除去前鰓亞綱螺的性畸形/性間性的特殊情況,在缺少特殊暴露標(biāo)志物的情況下,整體的反應(yīng)標(biāo)準(zhǔn)(存活率、生長(zhǎng)、生殖等)是目前評(píng)價(jià)無(wú)脊椎動(dòng)物內(nèi)分泌干擾物的唯一選擇.然而,這種方法很難確定對(duì)生物的影響是否歸因于化合物的內(nèi)分泌活性[41],所以,發(fā)展機(jī)制獨(dú)特性的生物標(biāo)志物在評(píng)價(jià)無(wú)脊椎動(dòng)物內(nèi)分泌干擾物測(cè)試中很有價(jià)值,同時(shí)生物標(biāo)志物還能用來(lái)指導(dǎo)實(shí)驗(yàn)物種的選擇[55].

5.4 拓展魚盤螺和銅銹環(huán)棱螺的實(shí)驗(yàn),整合福壽螺實(shí)驗(yàn)到國(guó)家標(biāo)準(zhǔn)程序中

下一步的研究應(yīng)該評(píng)價(jià)魚盤螺和銅銹環(huán)棱螺對(duì)培養(yǎng)條件的耐受性及其耐受性對(duì)生物量產(chǎn)率的影響,以及評(píng)價(jià)其他培養(yǎng)條件對(duì)產(chǎn)率的影響,進(jìn)而優(yōu)化培養(yǎng)條件.在此基礎(chǔ)上,研究?jī)煞N螺對(duì)大量污染物的敏感性信息.魚盤螺和銅銹環(huán)棱螺也可作為水體毒性受試生物,這需要進(jìn)一步實(shí)驗(yàn)無(wú)沉積物的情況.

提議將福壽螺作為巴西本土生物的代表,希望能將該生物常規(guī)用于除草劑和污水毒性的急性毒性測(cè)試中,并推薦將該毒性實(shí)驗(yàn)作為常規(guī)的程序列入國(guó)家標(biāo)準(zhǔn)程序中,作為早期預(yù)警檢測(cè)工具.

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Discussion on candidate species of gastropoda organisms in toxicity testing.

ZHENG Shi-mei1, ZHOU Qi-xing1,2*, XIONG Hong-xia1(1.Key Laboratory of Pollution Processes and Environmental Criteria, Ministry of Education, Tianjin Key Laboratory of Environmental Remediation and Pollution Control, College of Environmental Science and Engineering, Nankai University, Tianjin 300071, China；2.Key Laboratory of Terrestrial Ecological Process, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China). China Environmental Science, 2011,31(8)：1390～1397

With an attention to the hazard of endocrine disruptors on gastropoda organisms, the pollution-ecological problem of mollusks is being concerned gradually. Based on the current progress in the research area, several typical gastropoda organisms were discussed as the candidate species of testing organisms in the toxicity testing of chemicals. In view of their ecological significance, the four candidate prosobranchia species (Potamopyrgus antipodarum, Valvata piscinalis, Pomacea lineate and Bellamya aeruginosa) and one pulmonata organism (Lymnaea stagnalis) were suggested to develop as the standard testing organisms. Compared with the four other organisms proposed by foreign scholars, Bellamya aeruginosa proposed by Chinese scholars is of indigenous significance. Thus, attentions should be paid to the species in the toxicity testing of chemicals. On the basis of above discussions, the aspects of research on gastropoda organisms as candidate species in the toxicity testing in the future were prospected.

chemical；testing organism；gastropoda organism；pollution-ecological problem；ecological significance

X171.5

A

1000-6923(2011)08-1390-08

2010-11-01

國(guó)家環(huán)保公益性項(xiàng)目(200909041);國(guó)家自然科學(xué)基金資助項(xiàng)目(40930739)

* 責(zé)任作者, 教授, Zhouqx@nankai.edu.cn

鄭師梅(1983-),女,山東安丘人,博士研究生,主要從事水質(zhì)基準(zhǔn)與生態(tài)毒理學(xué)研究.發(fā)表論文3篇. adsorption and kinetics of chromium(VI) removal from aqueous solutions by Ocimum americanum L. seed pods [J]. J Hazard Mater, 2009,161:709-713.

作者簡(jiǎn)介：李國(guó)新(1981-),男,湖北公安人,博士,主要從事城市水環(huán)境生態(tài)與修復(fù)研究.發(fā)表論文10余篇.