Effects of p,p′-DDE exposure on gonadal development and gene expression in Japanese medaka （Oryzias latipes）

Although 1,1-dichloro-2,2-bis（p-chlorophenyl）-ethylene （p,p′-DDE）, the major and most persistent metabolite of dichlorodiphenyltrichloroethane （DDT）, was continually detected in wild fishes that showed abnormal gonad development such as intersex, little is known about the impact of p,p′-DDE exposure on gonad development in fishes. To survey the effects of p,p′-DDE on gonadal development and gene expressions, male juvenile （20-d post hatch） Japanese medaka （Oryzias latipes） was exposed to 1, 5, 20, and 100 μg/L p,p′-DDE for two months. Increased hepatosomatic index （HSI） and decreased gonadosomatic index （GSI） were found in the p,p′-DDE-treated groups. Intersex was found in 100 μg/L p,p′-DDE exposure group, as well as 100 ng/L 17α-ethynylestradiol （EE2） group. By quantitative real-time RT-PCR, it was found that gene expressions of vitellogenins （VTG-1, VTG-2）, choriogenins （CHG-H, CHG-L）, and estrogen receptor α （ER-α） in the liver of the fish were significantly up-regulated by p,p′-DDE exposure. VTG-1 and VTG- 2 were recommended as the preferred biomarker for assessing anti-androgenic p,p′-DDE because they were the highest up-regulated among the genes and showed good dose-response relationship. The up-regulated ER-α suggested that a potential synergetic effect would occur when p,p′-DDE coexists with other ER-α-binding endocrine-disrupting chemicals （EDCs）.     

柱孢藻毒素对草鱼淋巴细胞毒效应及氧化损伤机理研究

针对淡水水体拟柱孢藻水华产生的柱孢藻毒素,以我国典型食用鱼种草鱼(Ctenophar yngodon idellus)为实验材料,研究了其对鱼体免疫细胞毒效应及机理.结果表明,随着体外暴露剂量的增加,草鱼淋巴细胞活性逐渐降低,暴露24h后100μg·L-1柱孢藻毒素诱导组细胞活性仅为对照组的20%;对诱导组淋巴细胞的DNA检测发现呈现阶梯状电泳的典型细胞凋亡特征;应用流式细胞仪进一步检测了细胞凋亡率,表明1～100μg·L-1柱孢藻毒素均能够诱导细胞凋亡,并且其凋亡毒效应呈现明显的时间-效应和剂量-效应关系;诱导12h后,氧化应激产物活性氧(ROS)和丙二醛(MDA)含量均明显上升,100μg·L-1柱孢藻毒素诱导组ROS含量为对照组的2.5倍,MDA含量为对照组的2倍,诱导24h后,1、10、50和100μg·L-1实验组氧化应激产物含量仍然明显上升,这说明氧化应激是柱孢藻毒素导致草鱼淋巴细胞DNA损伤的重要原因;RT-PCR技术对重要凋亡基因的检测表明,1～100μg·L-1实验组凋亡促进基因Bax表达显著增强(p<0.05),50和100μg·L-1高剂量组凋亡抑制基因Bcl-2表达显著降低(p<0.01).本研究从细胞水平揭示柱孢藻毒素对草鱼免疫细胞具有明显的毒性,该毒效应通过氧化应激介导的DNA损伤表现,凋亡是柱孢藻毒素对草鱼免疫细胞毒性的主要机制.     

Effects of phenanthrene on hepatic enzymatic activities in tilapia (Oreochromis niloticus♂×O. aureus♀)

The effects of phenanthrene (Phe) on hepatosomatic index (HSI) and hepatic enzymatic activities in hybrid tilapia (Oreochromis niloticus ♀× O.aureus ♂) were investigated via the static freshwater exposure at dosage of 50,100 or 400 μg/L for 4-14 d.Compared with the control group,HSI was significantly decreased (P<0.05) at 400 μg/L at day 14.Increased enzymatic activities (P<0.05) were observed for catalase (CAT),glutathione peroxidase μgPx) and superoxide dismutase (SOD) at either 100 or 400 μg/L at day 8 and 14,as well as for CAT at 50 μg/L at day 14,except for GPx at 400 μg/L at day 8.Ethoxyresorufin O-deethylase (EROD) activity was significantly increased (P<0.05) at all dosage at day 4 as well as at 50 μg/L at day 8,but significantly decreased at either 100 or 400 μg/L at day 14 (P<0.05).Glutathione-S-transferase μgST) activity was not affected.The results suggest that CAT,GPx,SOD and EROD,as well as HSI in tilapia may be used as the biomarkers or indexes for evaluating or monitoring the pollution of polycyclic aromatic hydrocarbons (PAHs) such as Phe.     

五氯酚对稀有鮈鲫胚胎毒性效应研究

研究五氯酚(PCP)对稀有鮈鲫(Gobiocypris rarus)胚胎的致畸和毒性效应.以7.5,30,60,120,250μg/L5个浓度的PCP对0hpf(hpf,受精卵孵出时间)的稀有鮈鲫胚胎进行暴露染毒,同时设置空白对照组、二甲基亚砜溶剂对照组和雌二醇(EE2,2.5ng/L)阳性对照组.在立体显微镜下观察整个胚胎的发育过程,统计胚胎的孵化率、96hpf相对存活率和各时期的畸形率,并利用半定量RT-PCR检测胚胎中CYP1A基因和p53基因mRNA的表达.结果表明,PCP暴露能延迟稀有鮈鲫胚胎发育,并造成胚胎卵凝结、心包囊肿、脊柱弯曲等多种畸形甚至死亡.随着PCP暴露浓度的升高,稀有鮈鲫胚胎的孵化率和96hpf相对存活率降低,各时期的畸形率增加,并呈现一定的浓度效应.稀有鮈鲫胚胎CYP1A基因和p53基因mRNA表达被显著诱导,并随PCP浓度的升高而增加.PCP对稀有鮈鲫胚胎发育表现为显著的毒性效应.稀有鮈鲫胚胎孵化率、96hpf相对存活率、各时期畸形率及CYP1A基因和p53基因的诱导表达可以作为评价PCP毒性作用的敏感指标.     

壬基酚在鱼体组织中的累积及对鱼类性腺的影响

采用HPLC法测定了在一定条件下暴露后鲤鱼和草鱼体内壬基酚的含量和分布,分析了鱼类性腺重量的变化.结果表明,在试验浓度下,20～25d后鲤鱼和草鱼体内的壬基酚可达到累积和释放的动态平衡,其最大累积系数分别为84.3和78.8,累积规律为鲤鱼>草鱼,肝脏>肌肉.受试雌鱼的性腺重量有明显的提高,相当于100ng/L17β-雌二醇(E₂)作用下鱼类性腺重量的34%～41%,表明壬基酚对鱼类具有一定的雌激素活性,且壬基酚的生物累积性和雌激素活性在产生时间和活性数值方面均是同步的和成正相关的.     

四环素和砷对斑马鱼的联合毒性及机制

选取斑马鱼为模式生物,研究TET（四环素）和As（砷）对斑马鱼的联合毒性.结果表明,50μg/L TET和100μg/L As的联合暴露显著增加斑马鱼体内氧化应激（GPx）、炎症因子（TNF-α）和凋亡因子（C-jun）相关基因表达,造成肠道和肝脏组织更严重的损伤.50μg/L TET和100μg/L As联合暴露呈现的协同毒性效应与P-gp基因被抑制介导的As在肠道和肝脏蓄积量[（704.15±24.50） ng/g和（458.35±24.25ng/g）]显著增加有关.此外,发现肠道的通透性及微生物群落结构的变化对TET和As协同毒性无明显影响.     

氟对斑马鱼生殖毒性研究

为了探讨氟对鱼类的生殖毒性效应,以成年斑马鱼为试验对象,氟化钠暴露45 d和90 d后,运用组织切片观察斑马鱼性腺组织的发育情况,采用酶联免疫分析法测定性腺中雌二醇（Estradiol,E2）、卵黄蛋白原（Vitellogenin,VTG）和睾酮（Testosterone,T）的含量,并使用实时荧光定量PCR方法检测斑马鱼生殖相关基因的表达水平.结果显示：氟暴露会阻滞成年斑马鱼性腺发育,造成卵泡发育不良,精巢生精能力下降;同时降低雌性斑马鱼性腺中E2和VTG的含量,增加T的含量;雌性斑马鱼孕酮受体（Prg）和糖皮质激素受体（GR）基因相对表达量随氟浓度的增加呈先降后升的趋势,随着染毒剂量与时间的增加,雄激素受体（AR）基因相对表达量逐渐增加,而CYP19a基因相对表达量逐渐减少.雄性斑马鱼性腺中E2、VTG、T的含量及生殖相关基因表达的变化趋势则与雌性斑马鱼相反.综上所述,氟暴露可以通过影响斑马鱼体内性腺发育、E2、VTG、T含量及生殖相关基因表达来影响其生殖功能.实验结果可为进一步研究氟化物对水生动物的生殖毒性效应提供理论依据.     

Effects of phenanthrene on hepatic enzymatic activities in tilapia (Oreochromis niloticus♂×O. aureus♀)

The effects of phenanthrene (Phe) on hepatosomatic index (HSI) and hepatic enzymatic activities in hybrid tilapia (Oreochromis niloticus O. aureus ) were investigated via the static freshwater exposure at dosage of 50, 100, and 400 g/L for 4–14 d. Compared with the control group, HSI was significantly decreased (P < 0.05) at 400 g/L at day 14. Increased enzymatic activities (P < 0.05) were observed for catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) at either 100 or 400 g/L at day 8 and 14, as well as for CAT at 50 g/L at day 14, except for GPx at 400 g/L at day 8. Ethoxyresorufin O-deethylase (EROD) activity was significantly increased (P < 0.05) at all dosage at day 4 as well as at 50 g/L at day 8, but significantly decreased at either 100 or 400 g/L at day 14 (P < 0.05). Glutathione-S-transferase (GST) activity was not a ected. The results suggest that CAT, GPx, SOD and EROD, as well as HSI in tilapia may be used as the biomarkers or indexes for evaluating or monitoring the pollution of polycyclic aromatic hydrocarbons (PAHs) such as Phe.     

城市污水处理厂水样中有机氯农药残留分析

建立了基于HLB固相萃取柱和气相色谱/电子捕获(GC/ECD)分析城市污水中有机氯农药的分析方法,对北京市五大城市污水处理厂———高碑店、北小河、酒仙桥、清河、方庄污水处理厂进出水水样中有机氯农药进行了分析。在5个污水处理厂的水样中共检出了6种有机氯类农药,分别是α-六六六、β-六六六、γ-六六六、δ-六六六、艾氏剂、4,4’-滴滴伊和异狄氏剂,浓度在0.2~76.4ng/L之间。方法对有机氯农药的回收率达到60.93%~141.50%,方法检测限为(0.27~2.90)ng/L。     

乙炔基雌二醇对真鲷幼鱼的雌激素效应研究

采用静态暴露方式研究17α-乙炔基雌二醇(EE_2)对真鲷(Pagrosomus major)幼鱼的雌激素效应.当真鲷幼鱼暴露于0.01、0.1、0.5 μg/L EE_2,42 d后,真鲷幼鱼的肥满度极显著下降;血浆中卵黄蛋白原(Vtg)被诱导产生,肝胰脏指数(HSI)和血浆蛋白总量极显著升高;血浆雌二醇(E_2)和睾酮(T)水平显著降低,其中血浆雌二醇受EE_2的干扰更显著,降低的幅度更大.结果提示,真鲷幼鱼的肥满度、肝胰脏指数、血浆蛋白总量、血浆性激素水平等可作为评估EE_2等环境内分泌干扰物毒性效应的生物指标;真鲷幼鱼血浆中的Vtg具有作为生物标志物,用于监测海洋水体中EE_2等环境内分泌干扰物的潜力.     

Distribution and health risk assessment of organochlorine pesticides （OCPs） in industrial site soils： A case study of urban renewal in Beijing, China

A field survey was conducted in a contaminated industrial site of southern Beijing,China to investigate the contents and distribution of the organochlorine pesticides(α-,β-,γ-,δ-HCH,p,p-DDT,p,p-DDE,p,p-DDD and o,p-DDT) in the profiles of soil,and a health risk assessment was carried out with CalTOX multimedia exposure model.Results showed that mean concentrations of total hexachlorocyclohexane isomers(HCHs) and total dichlorodiphenyltrichloroethane isomers(DDXs) in soils were in the range of 13.20-148.71 mg...     

CuSO4对松江鲈肾细胞系的毒性效应

本文研究了不同浓度的CuSO₄对55-65代松江鲈肾细胞系的毒性效应。采用MTT法测得CuSO₄ 24 h LC₅₀为154.34 μmol/L。酶活力测定的结果显示:超氧化物歧化酶（SOD）和谷胱甘肽-S-转移酶（GST）的活性在CuSO₄浓度为0~300 μmol/L时,随浓度的升高逐渐升高,在300 μmol/L时达到最大值;谷胱甘肽过氧化物酶（GSH-Px）的活性在CuSO₄浓度为100 μmol/L时达到最大值,随后随着CuSO₄浓度的降低逐渐降低。微核率随CuSO₄浓度增加而增加,最高达14.33‰,彗星实验发现在半致死浓度条件下松江鲈肾细胞拖尾率为54.00%、迁移长度18.41±2.94 μm,与对照组差异显著（p ＜0.05）。认为CuSO₄会引起细胞氧化酶活性的改变以及细胞核DNA损伤。松江鲈肾细胞系可以作为Cu2+污染的监测指标。     

溴氰菊酯对稀有鮈鲫早期生命阶段发育和内分泌干扰毒性

采用半静态水体暴露方式研究了水中溴氰菊酯对稀有鮈鲫早期生命阶段的发育毒性与内分泌干扰效应.结果表明,影响稀有鮈鲫胚胎孵化的LOEC (最低可观察效应浓度)>3.0μg/L;影响稀有鮈鲫仔鱼发育畸形和死亡指标的LOEC和NOEC (无可观察效应浓度)分别为1.0和0.33μg/L.低至0.04μg/L的溴氰菊酯暴露便可显著下调稀有鮈鲫幼鱼体内雄激素受体基因(AR)表达量并上调甲状腺激素受体基因(TRβ)表达量;0.11μg/L的溴氰菊酯暴露可以下调稀有鮈鲫幼鱼体内雄激素受体基因(AR)、雌激素受体基因(ER1、ER2b)和芳香烃受体基因(AhR1a)的表达量,并上调甲状腺激素受体基因(TRβ)表达量;0.33μg/L的溴氰菊酯可以下调稀有鮈鲫幼鱼体内雄激素受体基因(AR)、雌激素受体基因(ER1、ER2b)、芳香烃受体基因(AhR1a、AhR1b、AhR2)的表达量,并诱导甲状腺激素受体基因(TRβ)表达量的上调.上述作用浓度水平已经处于多个天然水体中溴氰菊酯的检出浓度范围,因此,关于水体残留溴氰菊酯对鱼类的内分泌干扰效应必须予以重视.     

壬基酚聚氧乙烯醚对成年雄性斑马鱼HPG轴相关基因表达的影响

本研究以模式动物斑马鱼（Danio rerio）为受试生物,采用半静态水体暴露的方式研究了不同浓度壬基酚聚氧乙烯醚（NPEO）对斑马鱼雄性成鱼下丘脑-垂体-性腺轴（HPG轴）的影响.结果显示,考察浓度范围内的NPEO暴露可以显著上调斑马鱼脑中GnRH2、GnRHR1、GnRHR2、GnRHR4、FSHβ和LHβ基因,以及性腺中LHR基因的相对表达量.GnRHR和LHR基因表达量对较低浓度NPEO暴露较为敏感,其中GnRHR4和LHR基因表达量在低至0.001mg/L的NPEO暴露下即出现显著上调.0.1和10mg/L NPEO暴露可以显著抑制斑马鱼精巢中CYP17基因的表达量,而10mg/L NPEO暴露则可以显著诱导CYP19a基因的表达量.GnRH相关调控基因表达量的上调,表明NPEO暴露可以诱导下丘脑分泌GnRH,进而刺激垂体分泌GtH.NPEO暴露诱导CYP19a基因的表达,促进了内源雌激素的合成.同时,NPEO通过抑制CYP17表达,可能抑制睾酮（T）的合成,干扰斑马鱼精巢中原有的性激素平衡.斑马鱼精巢内雌激素水平升高负反馈给垂体,刺激垂体分泌促性腺激素.由此表明,考察浓度范围内（0.001~10mg/L）的NPEO的暴露可以影响雄性斑马鱼成鱼HPG轴的反馈调节.     

Toxic effects of crude-oil-contaminated soil in aquatic environment on Carassius auratus and their hepatic antioxidant defense system

Under the indoor simulant conditions, toxic effects of crude-oil-contaminated soil which was put into aquatic environment on the young fishes Carassius auratus and their hepatic antioxidant system after a 20-d exposure were investigated. Results showed that the relationship between the mortality of C. auratus and the exposed doses could be divided into 3 phases: fishes exposed to the low dose groups (0.5-5.0 g/L) were dead due to the ingestion of crude-oil-contaminated soils in aquatic environment; at the m...     

生物降解型消油剂与180#燃料油对黄海胆（Glyptocidaris crenularis）CAT和SOD活性的影响

为研究消油剂和溢油对海洋底栖生物的急性毒性效应,将黄海胆（Glyptocidaris crenularis）暴露于不同浓度（油水配比浓度分别为0.92 g/L、1.84 g/L、3.68 g/L）的180#燃料油分散液（water-accommodated fractions,WAFs）和经生物降解型消油剂处理后得到的乳化液（Biological enhanced water-accommodated fractions,BEWAFs）中,分别于暴露期24 h、48 h和96 h及恢复期24 h和48 h时,检测WAFs和BEWAFs对海胆性腺和肠组织的过氧化氢酶（CAT）和超氧化物歧化酶（SOD）的影响。结果显示,暴露在WAFs和BEWAFs中的性腺组织CAT、SOD和肠组织CAT活性总体呈先升高后降低趋势,而肠组织SOD活性随着暴露时间的增加呈升高—降低—升高趋势。肠组织中两种酶活性的最大诱导值出现的时间早于性腺组织,且恢复期时肠组织中各浓度组与对照组间差异更为显著,说明肠组织受石油烃氧化胁迫的敏感性比性腺组织强。与WAFs和BEWAFs相比,消油剂对照组中两种酶活性变化较小,由此可知,生物降解型消油剂本身对黄海胆的影响非常小,但其与燃料油对黄海胆的复合毒性效应却大于燃料油本身。海胆体内的CAT和SOD活性对石油烃的急性毒性效应较为敏感,可作为监测海上石油烃污染程度的生物标志物。     

Unexpected phenotypes of malformations induced in Xenopus tropicalis embryos by combined exposure to triphenyltin and 9-cis-retinoic acid

Xenopus tropicalis embryos were exposed for 48 hr to the mixtures of 5 g Sn/L triphenyltin(TPT), which is a well-known endocrine disruptor, and 0.25–5 g/L 9-cis retinoic acid(9c-RA), which is the natural ligand of retinoid X receptor. The phenotypes induced by combined exposure were more variable than those resulting from single exposure to either TPT or 9c-RA. The prominent phenotypes included underdeveloped head structures, abnormal eyes, narrow fins, enlarged proctodaeum, etc. Especially, combined exposure induced unexpected notochord malformations, which ranged from small swellings of the surface of the tails to the extension and extrusion of notochord out of the posterior tails. Compared with the 5 g Sn/L TPT-treated group, the index of fin deficiency was not affected, and the index of axis deficiency was significantly increased with increasing RA concentrations in the mixtures. Our results suggest that combined exposure to TPT and 9c-RA induced not only more variable phenotypes of malformations than exposure to single compound but also some new and unexpected phenotypes.     

DDTs在海河干流市区段沉积物/水间迁移行为研究

于2007年8月~2008年3月对海河干流表层水和沉积物中滴滴涕(DDTs)污染状况进行了采样调查.结果表明,海河干流水体DDTs含量为6.88~78.82 ng/L(均值28.54 ng/L),表层沉积物中DDTs含量为5.82~45.5 ng/g(均值21.55 ng/g),表层水和表层沉积物中DDTs主要成分分别为p,p-′DDT和p,p-′DDE.利用稳态非平衡逸度模型计算了p,p-′DDT在海河干流沉积物/水间迁移和分布,模型结果用现场实测浓度进行验证,计算值与实测值吻合很好.模型参数灵敏度分析进一步表明,生物降解速率常数、污染物在悬浮颗粒物/水间分配系数以及水体颗粒物沉降通量是影响p,p-′DDT在沉积物/水间迁移过程的主要因素.     

Prolonged manganese exposure induces severe deficits in lifespan, development and reproduction possibly by altering oxidative stress response in Caenorhabditis elegans

We examined the possible multiple defects induced by acute and prolonged exposure to high levels of manganese (Mn) solution by monitoring the endpoints of lifespan,development,reproduction,and stress response.Our data suggest that acute exposure (6 h) to Mn did not cause severe defects of life span,development,and reproduction.Similarly,no significant defects could be found for the life span,development,and reproduction in animals exposed to a low concentration of Mn (2.5 μmol/L) for 48 h.In contrast,prolonged exposure (48-h) to high concentrations of Mn (75 and 200 μmol/L) resulted in significant defects of life span,development,and reproduction,as well as the increase of the percentage of population with hsp-16.2::gfp expression indicating the obvious induction of stress responses in exposed animals.Moreover,prolonged exposure (48-h) to high concentrations (75 and 200 μmol/L) of Mn decreased the expression levels of antioxidant genes of sod-1,sod-2,sod-3,and sod-4 compared to control.Therefore,prolonged exposure to high concentrations of Mn will induce the severe defects of life span,development,and reproduction in nematodes possibly by affecting the stress response and expression of antioxidant genes in Caenorhabditis elegans.     

Using the nematode Caenorhabditis elegans as a model animal for assessing the toxicity induced by microcystin-LR

Among more than 75 variants of microcystin(MC),microcystin-LR(MC-LR) is one of the most common toxins.In this study,the feasibility of using Caenorhabditis elegans to evaluate MC-LR toxicity was studied.C.elegans was treated with MC-LR at different concentrations ranging from 0.1 to 80 μg/L.The results showed that MC-LR could reduce lifespan,delay development,lengthen generation time,decrease brood size,suppress locomotion behavior,and decreases hsp-16-2-gfp expression.The endpoints of generation time,brood...