苯并芘B[a]P对泥蚶组织EROD、GST酶活力和MDA含量的影响

采用实验生态学的方法,通过染毒和清除,研究了不同浓度(0.05、0.5、5和10 μg/L)苯并芘B[a]P胁迫15d和释放15d后,泥蚶(Tegillarca granosa)消化盲囊和鳃丝乙氧基异吩唑脱乙基酶(EROD)、谷胱甘肽转移酶(GST)活力和脂质过氧化物(MDA)含量的变化。结果表明: 在胁迫阶段0.5、5 和10μg/L,B[a]P处理组对泥蚶消化盲囊和鳃丝EROD、GST 酶活力和MDA含量显著影响(P＜0.05), EROD、GST 酶活力分别被诱导和抑制,第5d趋于稳定, MDA含量随时间呈上升趋势, 在第10d基本达到峰值并趋于稳定。在清除阶段, EROD活力和MDA含量逐渐下降, GST活力逐渐升高, 并在5～10d恢复到对照组水平。本研究中, EROD和GST活力的变化能够反映机体解毒代谢的能力, MDA含量的变化反映了机体氧化损伤的程度,表现出了一定的剂量和时间效应。     

苯并(a)芘影响黑鲷肝脏EROD活性变化的动力学研究

研究了黑鲷(Sparusmacro cephalus)在实验生态条件下暴露于不同浓度苯并(a)芘(benzo(a)pyrene,BaP)后,肝脏乙氧基异吩噁唑酮脱乙基酶(EROD)活性在不同暴露时相的动力学变化情况.EROD活性采用动力学方法测定.实验结果表明,暴露于较低质量浓度BaP(0.5,1.0μg/L)黑鲷肝脏的EROD活性在2 d后极显著高于对照组(P<0.01),2.0μg/L组则在2 h后极显著高于对照组,并且随暴露时间的延长,诱导程度表现出不同的下降趋势,而暴露于高质量浓度组(5.0 μg/L)的肝脏EROD活性极显著性诱导出现在12h.总体上较高质量浓度组(2.0,5.0μg/L)EROD活性出现极显著性诱导的时间比低质量浓度组(0.5,1.0μg/L)早,诱导程度也比低质量浓度组的高.在净化实验中各暴露组肝脏EROD活性均下降,与空白对照组比较均没有显著性差异,说明试验浓度没有超出严重损伤肝脏自身恢复系统的范围,在此浓度范围内污染的鱼通过净化可清除BaP污染.     

环境因子对栉孔扇贝血淋巴中谷胱甘肽转硫酶活性的影响

采用分光光度法研究了环境因子对栉孔扇贝（Chlamys farreri）血淋巴中谷胱甘肽转硫酶（GST）活性的影响。结果表明,血清和血细胞中都存在GST,且血细胞中酶活力高于血清。血细胞中GST的适宜温度范围是20～35℃,最适温度为30℃,在60℃时酶活力完全丧失。该酶的适宜pH为7—8,最适pH为7．5。将血细胞裂...     

基于综合生物标志物响应评价苯并芘对脊尾白虾(Exopalaemon carinicauda)的毒性效应

苯并[a]芘(Ba P)是一类典型多环芳烃(PAHs)污染物。为了定量评价Ba P对脊尾白虾(Exopalaemon carinicauda)的毒性效应,采用水体暴露实验,研究了脊尾白虾肝胰脏、肌肉组织的5种生物标志物(EROD、GST、SOD、CAT、MDA)对Ba P暴露的响应,并应用综合生物标志物响应(IBR)定量评价了Ba P对肝胰脏、肌肉组织的毒性差异。结果显示:肝胰脏、肌肉组织的EROD、GST、SOD、CAT活性整体呈现诱导-抑制的规律性变化,除了0.05μg/L实验组CAT外,其余各实验组肝胰脏组织生物标志物活性在实验结束时均显著高于对照组(P0.05),除了0.45μg/L实验组EROD、CAT外,其余各浓度组肌肉组织生物标志物活性在实验结束时均与对照组无显著性差异(P0.05),MDA含量呈现持续上升的趋势,在实验结束时各实验组均显著高于对照组(P0.05);IBR值存在时间效应和剂量效应。研究结果表明,肝胰脏、肌肉组织对Ba P的暴露响应存在差异性,肝胰脏生物标志物综合应激活性高于肌肉组织。     

海水酸化和磺胺甲恶唑复合暴露对大马蹄螺抗氧化酶活性和脂质过氧化的影响

为探讨海水酸化和磺胺甲恶唑环境残留对珊瑚礁区生物的影响,本文以南海珊瑚礁区关键礁栖生物大马蹄螺（Trochus niloticus） 为研究对象,探究了两种环境胁迫因素对大马蹄螺的氧化胁迫作用。分别采用两个 pH 水平 （pH 8.1、 pH 7.6） 和三个磺胺甲恶唑浓度水平 （0 ng/L、100 ng/L 和 1 000 ng/L） 单独和复合暴露 28 天,测定其对大马蹄螺鳃组织的超氧化物歧化酶 （SOD）、过氧化氢酶 （CAT） 和谷胱甘肽巯基转移酶 （GST） 活性以及谷胱甘肽 （GSH） 含量和脂质过氧化水平 （LPO） 的影响。结果发现：海水酸化和磺胺甲恶唑单独暴露能够导致大马蹄螺鳃组织出现不同程度的氧化应激现象,表现为抗氧化酶 （CAT、SOD 和 GST） 活性和谷胱甘肽含量的升高。在复合暴露条件下,大马蹄螺鳃组织的抗氧化酶（CAT、SOD 和 GST） 活性和谷胱甘肽含量均受到不同程度的抑制,同时伴随着脂质过氧化水平的显著升高。这表明短期海水酸化和磺胺甲恶唑复合暴露可导致大马蹄螺鳃组织出现氧化损伤现象,进而可能对个体存活和种群繁衍造成负面影响。     

原油WSF对三种海洋鱼类肝微粒体EROD活性的诱导和恢复的比较

研究了实验室条件下原油水溶性组分(WSF)暴露对黑鲷、黄鳍鲷和褐菖鲉肝微粒体EROD活性的剂量-效应,时间-效应和恢复过程。实验结果表明,在剂量诱导实验中,褐菖鲉肝EROD活性在原油WSF浓度为50μg/dm3时呈现生物统计学上的显著差异,而黑鲷和黄鳍鲷肝EROD活性在75μg/dm3时才呈现生物统计学上的显著差异;褐菖鲉肝EROD活性诱导倍数最高,但黑鲷的诱导浓度范围较广。在时间诱导实验中,在40μg/dm3原油WSF暴露下黄鳍鲷肝EROD活性在2 d时首先呈现显著差异;三种鱼肝EROD活性均在第4天达到最高,并呈现显著性变化,此后随着暴露时间的延长而逐渐下降并接近对照组水平。在恢复实验中三种鱼肝EROD活性下降并恢复到对照组水平。研究结果表明:对于石油污染物,黑鲷、黄鳍鲷和褐菖鲉肝EROD活性都可以作为污染生化效应监测指标,然而就三种鱼类比较而言,褐菖鲉最敏感,更适合于作为石油类污染及其生化效应,尤其是低剂量效应的监测生物。     

不同浓度苯并（a）芘对杂色鲍抗氧化酶活性的影响

为阐释不同浓度苯并（a）芘[B（a）P]对杂色鲍（Haliotis diversicolor）的毒性效应,以0.02、0.04和0.08 mg/dm^33个质量浓度的B（a）P对杂色鲍进行水体暴露胁迫,7d后检测肌肉、外套膜、鳃、性腺、肾和肝胰腺的超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽S-转移酶（GST）和谷胱甘肽过氧化物酶（GPx）活性及还原型谷胱甘肽（GSH）含量.结果显示：0.02 mg/dm^3的B（a）P暴露对杂色鲍抗氧化系统的影响不明显,而0.04、0.08 mg/dm^3的B（a）P则显著抑制了SOD、CAT和GPx酶活性以及GSH含量,同时对GST酶活性具有显著的诱导作用;实验还发现,不同组织抗氧化酶活性对B（a）P胁迫的敏感性响应存在较大差异,其中鳃、肾脏和肝胰腺的敏感性响应明显高于肌肉和外套膜.上述结果表明,苯并（a）芘暴露对杂色鲍抗氧化酶活性的影响具有明显的剂量-效应关系及组织差异性,杂色鲍的鳃、肾脏或者肝胰腺的抗氧化酶响应多环芳香烃胁迫的更敏感,可作为B（a）P污染的生物标志物.     

As（Ⅲ） 暴露对紫贻贝抗氧化酶活性和 脂质过氧化的影响

为探讨As（Ⅲ）对紫贻贝（Mytilus galloprovincialis） 肝胰腺和鳃组织抗氧化能力的影响,测定了1、10、100 ug/L As （Ⅲ） 暴露30 天后,肝胰腺和鳃组织超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽巯基转移酶（GST）的活性 变化以及谷胱甘肽（GSH） 和丙二醛（MDA） 含量的变化。结果发现：As（Ⅲ）暴露对紫贻贝肝胰腺CAT、SOD、GST 活性 以及GSH 含量无显著影响,高浓度As（Ⅲ） 暴露可诱导MDA含量的显著升高;对鳃组织而言,中等剂量暴露组CAT 活性 以及GSH含量较对照组水平极显著升高,高剂量As（Ⅲ） 暴露组MDA 含量较对照组也显著升高。研究结果表明,低浓度 As （Ⅲ） 长期暴露即可导致紫贻贝产生氧化胁迫,导致肝胰腺和鳃组织的脂质过氧化,且As（Ⅲ） 对紫贻贝鳃组织的影响 较肝胰腺组织更加显著。     

原油WSF对三种海洋鱼类肝微粒体EROD活性的诱导和恢复的比较

研究了实验室条件下原油水溶性组分(WSF)暴露对黑鲷、黄鳍鲷和褐菖鲉肝微粒体EROD活性的剂量-效应,时间-效应和恢复过程。实验结果表明,在剂量诱导实验中,褐菖鲉肝EROD活性在原油WSF浓度为50 μg/dm³时呈现生物统计学上的显著差异,而黑鲷和黄鳍鲷肝EROD活性在75 μg/dm³时才呈现生物统计学上的显著差异;褐菖鲉肝EROD活性诱导倍数最高,但黑鲷的诱导浓度范围较广。在时间诱导实验中, 在40 μg/dm³原油WSF暴露下黄鳍鲷肝EROD活性在2 d时首先呈现显著差异; 三种鱼肝EROD活性均在第4天达到最高,并呈现显著性变化,此后随着暴露时间的延长而逐渐下降并接近对照组水平。在恢复实验中三种鱼肝EROD活性下降并恢复到对照组水平。研究结果表明:对于石油污染物,黑鲷、黄鳍鲷和褐菖鲉肝EROD活性都可以作为污染生化效应监测指标,然而就三种鱼类比较而言,褐菖鲉最敏感,更适合于作为石油类污染及其生化效应,尤其是低剂量效应的监测生物。     

黄芩苷对中国对虾细胞色素P450酶及谷光甘肽-S-转移酶活性的影响

用紫外分光光度法及荧光分光光度法测定中国对虾肝胰腺及鳃氨基比林-N-脱甲基酶(APND)、乙氧基香豆素-O-脱乙基酶(ECOD)、谷光甘肽-S-转移酶(GST)几种药物代谢酶的活性,探讨黄芩苷对上述几种酶活性的影响.实验结果表明,中国对虾肝胰腺和鳃丝APND活性分别为3.172和2.242 nmol/(min·mgprotein~(-1))、ECOD活性分别为0.325 0和0.357 9 pmol/(min·mgprotein-~(-1))、GST活性分别为70.59和13.97 nmol/(min·mgprotein~(-1)).中国对虾连续7 d投喂黄芩苷药饵(100 mg/kg)后,黄芩苷对几种酶均有不同程度的诱导作用,酶的活性均呈现先升高后降低的趋势.肝胰腺和鳃丝APND活性均在3 h左右达到最高,ECOD活性均在1 h左右达到最高,而GST活性迟于CYP450酶被诱导,2种组织GST活性分别在6,12 h达到最高水平.在临床用药中应考虑到黄芩苷对药物代谢酶的诱导作用,适当用药,合理配伍.     

Effects of β-naphthoflavone on hepatic biotransformation and glutathione biosynthesis in largemouth bass (Micropterus salmoides)

We are investigating the effects of in vivo exposure of prototypical enzyme inducing agents on hepatic biotransformation enzyme expression in largemouth bass (Micropterus salmoides), a predatory game fish found throughout the United States and Canada. The current study targeted those genes involved in biotransformation and oxidative stress that may be regulated by Ah-receptor-dependent pathways. Exposure of bass to β-naphthoflavone (β-NF, 66 mg/kg, i.p.) elicited a 7–9-fold increase in hepatic microsomal cytochrome P4501A-dependent ethoxyresorufin O-deethylase (EROD) activities, but did not affect cytosolic GST catalytic activities toward 1-chloro-2,4-dinitrobenzene (CDNB) or 5-androstene-3,17-dione (ADI). Glutathione S-transferase A (GST-A) mRNA expression exhibited a transient, but non-significant increase following exposure to β-NF, and generally tracked the minimal changes observed in GST–CDNB activities. Expression of the mRNA encoding glutamate-cysteine ligase catalytic subunit (GCLC), the rate-limiting enzyme in glutathione (GSH) biosynthesis, was increased 1.7-fold by β-NF. Changes in GCLC mRNA expression were paralleled by increases in intracellular GSH. In summary, largemouth bass hepatic CYP1A-dependent and GSH biosynthetic pathways, and to a lesser extent GST, are responsive to exposure to β-NF.     

Biomarker responses to pollution in two invertebrate species: Scrobicularia plana and Nereis diversicolor from the Cádiz bay (SW Spain)

The clam Scrobicularia plana and the polychaete worm Nereis diversicolor were collected in several sites from a littoral enclosure in SW Spain. The aim of our study was to relate various biomarker responses in these species to a pollution gradient caused by untreated domestic discharges and to verify the adequacy of the selected species as sentinels in this habitat. The biomarkers selected were the antioxidant enzymes catalase (CAT), glutathione peroxidase (GPX) and DT-diaphorase (DT-D). In addition, the activities of cytochrome P450-dependent ethoxyresorufin O-deethylase (EROD) activity, the phase II detoxifying enzyme glutathione S-transferase (GST) and the neurotoxicity marker acetylcholinesterase (AChE) were measured. Metallothionein levels were selected as biomarkers of heavy metals exposure in both species. The results suggest a different response in the water filtering organism (clam) and the sediment eater (polychaete), probably as a consequent of different pollution exposure and that samples from the “Caño Sancti-Petri” were exposed to biologically active compounds that altered some of their biochemical responses. AChE was the most sensitive biomarker in both species and N. diversicolor proved to be a more robust sentinel in this ecosystem.     

Short-term variability of multiple biomarker response in fish from estuaries: Influence of environmental dynamics

Short-term variability in biomarker responses and the effects of temperature and salinity variation were explored in three fish species (Dicentrarchus labrax, Solea senegalensis and Pomatoschistus microps) occurring in the Tejo estuary. Short-term variability in biomarkers was observed in all species although no pattern was discerned over time (days to weeks). Antioxidant enzymes activity (superoxide dismutase (SOD) and catalase (CAT)) showed low temporal variability, indicating some constancy or baseline level in antioxidant responses. Only CAT activity in S.?senegalensis was correlated with temperature, suggesting that exposure to contaminants triggered antioxidant acclimation. Higher short-term variability was observed in xenobiotic biotransformation enzymes activity (phase I ethoxyresorufin O-deethylase (EROD) and phase II glutathione S-transferase (GST)). Yet a significant correlation between EROD and GST in D.?labrax and S.?senegalensis suggests a concomitant response to contaminants. Moreover the lack of correlation between xenobiotic biotransformation enzymes and abiotic variables on concordant time scales, suggest a high specificity of these biomarkers to chemical exposure, rather than high variability due to environmental dynamics.     

三氯异氰尿酸对脊尾白虾幼体存活及发育的影响

三氯异氰尿酸(Trichloroisocyanuric acid ,TCCA)是水产养殖中常用的水质消毒剂, 使用不当会给生产和环境带来危害。为指导脊尾白虾(Exopalaemon carinicauda)人工育苗过程中TCCA 的安全用药,作者利用静水试验法研究了TCCA 对脊尾白虾幼体的急性毒性及对幼体发育的影响。实验设7 个浓度梯度组(1.0、1.5、2.0、2.5、3.0、3.5、4.0 mg/L)和1 个空白对照组, 每组3 个平行。急性毒性实验持续96 h 后结束, 用正常育苗海水代替TCCA 溶液, 正常管理直到幼体全部变态为仔虾, 评估短时间的TCCA 处理对脊尾白虾幼体发育的影响。结果表明, TCCA 对脊尾白虾幼体在24、48、72 和96 h 的半致死浓度分别为3.81、3.03、2.71 和2.47 mg/L, 其安全浓度为0.25 mg/L; 随着TCCA 浓度的升高, 脊尾白虾幼体变态存活率显著降低(P<0.05), 但能成功变态为仔虾个体的变态持续时间及仔虾体长与对照组无明显差异(P>0.05)。本实验结果可以为脊尾白虾人工育苗安全用药提供参考和依据。     

Effects of beta-naphthoflavone on hepatic biotransformation and glutathione biosynthesis in largemouth bass (Micropterus salmoides)

We are investigating the effects of in vivo exposure of prototypical enzyme inducing agents on hepatic biotransformation enzyme expression in largemouth bass (Micropterus salmoides), a predatory game fish found throughout the United States and Canada. The current study targeted those genes involved in biotransformation and oxidative stress that may be regulated by Ah-receptor-dependent pathways. Exposure of bass to beta-naphthoflavone (beta-NF, 66 mg/kg, i.p.) elicited a 7-9-fold increase in hepatic microsomal cytochrome P4501A-dependent ethoxyresorufin O-deethylase (EROD) activities, but did not affect cytosolic GST catalytic activities toward 1-chloro-2,4-dinitrobenzene (CDNB) or 5-androstene-3,17-dione (ADI). Glutathione S-transferase A (GST-A) mRNA expression exhibited a transient, but non-significant increase following exposure to beta-NF, and generally tracked the minimal changes observed in GST-CDNB activities. Expression of the mRNA encoding glutamate-cysteine ligase catalytic subunit (GCLC), the rate-limiting enzyme in glutathione (GSH) biosynthesis, was increased 1.7-fold by beta-NF. Changes in GCLC mRNA expression were paralleled by increases in intracellular GSH. In summary, largemouth bass hepatic CYP1A-dependent and GSH biosynthetic pathways, and to a lesser extent GST, are responsive to exposure to beta-NF.     

Ethoxyresorufin-O-deethylase activity and fixed wavelength fluorescence detection of PAHs metabolites in bile in turbot (Scophthalmus maximus L.) Exposed to a dispersed topped crude oil in a Continuous Flow System

Long term effects of sublethal concentrations of oil on the marine environment have become of general concern. Cytochrome P4501A activity (EROD) in liver and fixed wavelength fluorescence detection of PAHs metabolites (FF) have in this study been used as biomarkers for dispersed oil exposure on a long term period of juvenile turbot (Scophthalmus maximus L.). A Continuous Flow System was used to carry out the study. The fish were continuously exposed to 0.125, 0.5 or 2.0 mg litre⁻¹ dispersed topped crude oil for 6, 15, 24 h, 4 and 21 days followed by a 9 days recovery period in clean seawater. No induction of the cytochrome P4501A was measured. A maximum level in bile metabolites (4- to 5-fold) was recorded after 24 h of exposure revealing thereby a detoxification process, but a decline occurred from day 4 to day 21. This study demonstrated that FF detection of PAHs metabolites in bile could be a more sensitive biomarker than EROD activity in a long term exposure to sublethal concentration of oil.     

Cytochrome P4501A1 induction in Kenai River sculpin (Cottus spp.) as a monitor of freshwater pollution effects

Freshwater sculpin were taken from various locations in the Kenai River to determine their in-vivo response to pollution exposure. The specific activity of cytochrome P4501A1 was determined using fluorescence detection of the 7-ethoxyresorufin-O-dealkylase (EROD) reaction. The EROD specific activity was found to be independent of river miles but appeared to relate to specific sites of urban runoff. Glutathione-S-transferase (GST) activities were also measured but showed no significant difference between sites. Cytochrome P450 laboratory induction studies were carried out on individuals collected from a pristine site. Five groups of 10 fish were dosed from 10–150 mg β-Naphthoftavone (βNF)/kg body weight. The response was linear from 10–50 mg βNF/kg body weight. Sculpin appear to have excellent potential as a sensitive indicator of xenobiotic exposure in a freshwater benthic habitat.     

Pollutant-induced cell injury in fish liver: Use of fluorescent molecular probes in live hepatocytes

This study was conducted to test whether live cells from the liver of fish could be used to detect early changes that are indicative of pollutant-induced liver damage. Low-molecular-weight fluorescent molecular probes were inserted into isolated hepatocytes from dab (Limanda limanda) from a contaminated site and a reference site in the North Sea. These included bioprobes for endoplasmic reticulum (ER), cytochrome P-450-associated 7-ethoxyresorufin-O-deethylase (EROD), oxyradicals, reduced glutathione (GSH) and microtubules (MT). Endocytosis of Texas Red-albumin was used as an integrated indicator of hepatocyte performance. Findings showed increases in ER-associated fluorescence, EROD and oxyradical generation, with a marked decrease in endocytosis in hepatocytes from fish caught at the contaminated site. These results indicate that fish from the historically contaminated site were impacted by organic xenobiotics which induced the biotransformation system, radical production and cell injury.     

Biomarker responses to pollution in two invertebrate species: Scrobicularia plana and Nereis diversicolor from the Cádiz bay (SW Spain)

The clam Scrobicularia plana and the polychaete worm Nereis diversicolor were collected in several sites from a littoral enclosure in SW Spain. The aim of our study was to relate various biomarker responses in these species to a pollution gradient caused by untreated domestic discharges and to verify the adequacy of the selected species as sentinels in this habitat. The biomarkers selected were the antioxidant enzymes catalase (CAT), glutathione peroxidase (GPX) and DT-diaphorase (DT-D). In addition, the activities of cytochrome P450-dependent ethoxyresorufin O-deethylase (EROD) activity, the phase II detoxifying enzyme glutathione S-transferase (GST) and the neurotoxicity marker acetylcholinesterase (AChE) were measured. Metallothionein levels were selected as biomarkers of heavy metals exposure in both species. The results suggest a different response in the water filtering organism (clam) and the sediment eater (polychaete), probably as a consequent of different pollution exposure and that samples from the "Ca?o Sancti-Petri" were exposed to biologically active compounds that altered some of their biochemical responses. AChE was the most sensitive biomarker in both species and N. diversicolor proved to be a more robust sentinel in this ecosystem.     

Monitoring of the chemical pollution of the river Rhône through measurement of DNA damage and cytochrome P4501a induction in chub (Leuciscus cephalus)

The in vivo response of freshwater fish exposed to pollutants was assessed using two biomarkers, 7-ethoxyresorufin-O-deethylase (EROD) induction and DNA single strand breaks. Chub (Leuciscus cephalus) were caught in spring and in fall at various locations in the river Rhône watershed. EROD activity was measured in the liver while DNA damage was evaluated in chub erythrocytes using the recently developed Comet assay. Chemical contamination was evaluated both in fish muscle (PCBs) and in sediment (PCBs, PAHs, heavy metals) collected at each sampling station. Sex of individuals was shown to influence the level of EROD activity but not the level of DNA damage. The EROD activity as well as the DNA damage were found to be higher in the mostly contaminated stations compared to the reference one. This study shows that multibiomarker-based approach provides complementary informations about early effects in feral fish exposed to complex chemical pollution and highlights the interest of the Comet assay in genotoxicity assessment.