Relation of hepatic EROD activity and CYP1A level in Sebastiscus marmoratus exposed to benzo[a]pyrene

This study was designed to investigate the in vivo effects of benzo[a]pyrene (BaP) on hepatic ethoxyresorufin-O-deethylase (EROD) activity and its correlation with cytochrome P450 1A (CYP1A) protein levels in Sebastiscus marmoratus, which were exposed through a water column to BaP (10, 100, 1000 ng/L, respectively) or were treated with intraperitoneal injections of BaP (0.5, 1, 5, 10 mg/kg, respectively) every 7 d. The results showed that after 25 d of waterborne exposure to 1000 ng/L BaP, fish hepatic CYP1A levels and EROD activity were significantly induced. In contrast, EROD activity was not altered 7 d after second ip injections, whereas, CYP1A protein levels were increased. Dose-dependent increase of biliary BaP metabolites demonstrated that the catalytic activity of CYP1A was induced by treatment with BaP. The lowest observable effect concentration with regard to biliary BaP metabolites (100 ng/L) was much lower than that with reference to EROD activity (1000 ng/L). The results suggest that biliary polycyclic aromatic hydrocarbon (PAH) metabolites were shown to better reflect the contamination gradients of PAHs than EROD activity. It appeared to be necessary to measure CYP1A protein levels to complement the EROD activity in relevant toxicological assessments.     

Bioaccumulation and the expression of hepatic cytochrome P450 genes in marine medaka (Oryzias melastigma) exposed to difenoconazole

This study was conducted to assess the effects of difenoconazole (DFZ), a triazole fungicide, on the hepatic biotransformation system and its bioaccumulation in marine medaka (Oryzias melastigma). Fish were exposed to DFZ (1, 10, 100, 1000 ng/L) for 180 days. The results showed that: (1) The mRNA levels of hepatic CYP1A1, CYP1B, CYP1C1, CYP27B and CYP3A40 were up-regulated, but those of CYP3A38 and CYP27A1 were down-regulated. (2) The activity of ethoxyresorufin-O-deethylase (EROD) and the content of reduced glutathione (GSH) in the liver were increased in the DFZ-treated groups, and glutathione S-transferase (GST) activity was increased in the 100 and 1000 ng/L groups. (3) DFZ was accumulated in the muscle and the biological concentration factors in the 10, 100, and 1000 ng/L groups were respectively 149, 81 and 25. These results suggested that long-term exposure to DFZ at low concentrations would result in a bioaccumulation of this compound and disturb the biotransformation system.     

苯并[a]芘对鲫鱼生物标志物的影响研究

研究了苯并[a]芘暴露对鲫鱼(Carassius auratus)的几种生物标志物的影响.在鲫鱼接受3个不同质量分数苯并[a]芘(BaP)的腹腔注射14 d后,分析了其肝脏7-乙氧基-3-异吩噁唑酮-脱乙基酶(EROD)活性,以及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)等抗氧化酶活性和红细胞核异常(NA)率变化.结果显示:w(BaP)为1 mg/kg处理组的EROD活性显著升高(P＜0.001).w(BaP)为0.1和1 mg/kg处理组SOD活性显著低于对照组(P＜0.001),在暴露结束时,w(BaP)为1 mg/h处理组CAT活性显著低于对照组(P＜0.01),而w(BaP)为0.1和1 mg/kg处理组GPx活性被显著抑制(P＜0.001).表明BaP对EROD活性有诱导作用,而对SOD,CAT和GPx等抗氧化酶活性的抑制作用明显.所有处理组NA率与对照组均无显著差异.因此,鲫鱼EROD,SOD,CAT和GPx等酶活性的改变适于作为生物标志物,可以用来显示暴露于多环芳烃有机污染物鱼类的生活状态.      

Ploidy-, gender-, and dose-dependent alteration of selected biomarkers in Clarias gariepinus treated with benzo[a]pyrene

Naturally-occurring and artificially-induced polyploids have been documented in various fish species but to date no comparison has been reported of the impacts of ploidy on fish biomarker responses to organic pollutants. This study describes effects of ploidy, gender, and dose on biliary fluorescent aromatic compound (FAC) concentrations, hepatic ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST) activities in one of the most commonly cultured warm-water species, the African catfish Clarias gariepinus. Recently matured male and female diploid and triploid fish were intraperitoneally (i.p.) injected with 0, 5 or 25 mg/kg benzo[a]pyrene (BaP) and liver and gallbladder were sampled 48 hr later. No significant differences were found between ploidies in bile concentrations of 7,8 dihydrodiolbenzo[a]pyrene (7,8D BaP), 1-hydroxybenzo[a]pyrene (1-OH BaP) or 3-hydroxybenzo[a]pyrene (3-OH BaP). However, concentrations of the biliary FACs did differ between males and females at different dose of injection with generally higher concentrations in females at the low dose of BaP and higher concentrations in males at the higher BaP concentration. Hepatic EROD activity did not exhibit gender-dependent difference, whereas it was significantly higher in triploids than diploids. GST activities were not significantly influenced by any of the tested factors. This work advanced our understanding of the role of ploidy, gender, and dose in biotransformation of pollutants in fish.     

Combined effect of tributyltin and benzo[a] pyrene on the levels of sex hormone and vitellogenin in female Sebastiscus marmoratus

Tributyltin（TBT）, an organometal used as an antifouling biocide, has been reported to induce masculinization of fish. Benzo [a]pyrene （BaP）, a widespread carcinogenic polycyclic aromatic hydrocarbon, has been reported that its microsomal metabolites can produce an estrogenic response when tested in vitro. This study was therefore designed to examine the potential in vivo influence of TBT, BaP and their mixture on sex hormone levels in serum of Sebastiscus marmoratus, which were given 2 separate intraperitoneally （ip） injections（a single injection every 7 d） of TBT（0.5, 1, 5 and 10 mg/kg）, BaP（0.5, 1, 5 and 10 mg/kg）, or both in combination（0.5, 1, 5 and 10 mg/kg）; control fish received olive oil vehicle only. Six days after the 2nd injection, serum samples were collected and analyzed for sex hormone levels and alkali labile protein phosphorus （ALPP）, which is related to the yolk precursor protein vitellogenin. The pollutants at all doses significantly reduced serum testosterone, estradiol and ALPP content after 2 injections compared with the corresponding controls. The reduction of the estradiol levels should be response for the decrease of the vitellogenin levels. The results in the present study suggested that aromatase seems not the major target acted by TBT and BaP in fish. This study demonstrated that TBT or BaP exposure both inhibit the reproductive potential in female Sebastiscus marmoratus. Combined effect of TBT and BaP on the serum testosterone, estradiol and ALPP was not antagonism from the anticipation.     

鲫鱼肝脏中CYP1A1的诱导作为沉积物中二的毒理学指标的研究

在实验室内用鲫鱼分别暴露于含有PCDD/Fs和PCBs严重污染的沉积物和未受污染的沉积物中,测定鱼肝脏中细胞色素P4501A1（CYP1A1）依赖的7－乙氧基－3－异吩唑酮－脱乙基酶（EROD）的活性。结果表明,鲫鱼暴露于受二类化合物污染沉积物中1周后,肝脏中EROD的活性是对照组的83倍,而未被污染的沉积物暴露试验中EROD的活性和对照组没有明显差别。在此沉积物－水实验系统中二类污染物的TCDD等当量浓度（TEQ/L）与肝脏EROD的活性有较好的相关性。较长时间暴露,得到肝脏EROD的活性随时间的变化关系。还获得了试验温度对EROD活性诱导的影响。此实验表明,用实验室鲫鱼暴露于沉积物－水系统中,鱼肝脏EROD活性诱导可作为沉积物毒性评价指标,用于受二类化合物污染的沉积物样品的筛选,并有希望成为此类污染物的半定量检测方法。     

京津冀民用燃煤减排对大气中BaP污染影响评估

运用空气质量模型CMAQ模拟评估民用燃煤减排政策对京津冀大气中BaP污染状况的改进效果,模拟包括作为基准情景的2014年和低消减和高消减2个减排情景.模拟结果表明：基准情景下,京津冀BaP年均浓度为2.54ng/m³,超过国家空气质量标准（1ng/m³）,呈现1月 > 4月 > 10月 > 7月的季节变化特点,反映出冬季供暖燃煤的影响;京津冀南部BaP浓度高于北部,推测原因是南部BaP排放量较高.1月削减民用燃煤排放量对降低该地区BaP浓度和沉降量效果最显著,低削减和高削减下,北京、保定、廊坊BaP浓度分别比基准情景降低30%和40%以上;4、7、10月削减民用燃煤排放量对该地区BaP浓度和沉降量的变化影响不大.京津冀民用燃煤联防联控能更加有效地降低该地区BaP浓度.     

室内空气中苯并[a]芘的健康限值研究

应用健康风险评估方法,基于中国人群暴露参数特征推导我国不同人群的室内苯并[a]芘（Benzo[a]pyrene,BaP）浓度限值.并通过文献调研收集我国室内空气中BaP浓度,运用蒙特卡洛方法计算不同人群的BaP暴露致癌风险,探讨限值的可达性.研究发现,基于具有全国代表性人群暴露特征推导所得的BaP限值的平均值±标准偏差为（1.38±0.32）ng/m³,与我国现行《室内空气质量标准》（GB18883-2002）中的BaP限值相近.基于城市男性、女性暴露特征推导的健康限值分别为1.34ng/m³和1.40ng/m³,基于农村男性、女性暴露特征推导的限值分别为1.35ng/m³和1.41ng/m³.基于低年龄人群暴露特征推导的限值要低于高年龄段人群限值.我国大部分室内空气BaP实际监测浓度高于本研究推导出的健康限值,建议持续加强室内清洁能源使用和控烟等措施,减少BaP室内暴露风险.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

苯并(a)芘致毒的鱼的分子生态毒理学指标研究

通过测定典型的多环芳烃类物质苯并（a）芘（BaP）致毒后鱼体内几种重要分子生态毒理学指标的变化,来反映苯并（a）芘致毒对鱼体的影响．结果表明,肝脏ATPase活性降低,GST活性升高,DNA加合物相对标记水平（RAL）也增大,而EROD活性没有明显改变．这说明苯并（a）芘致毒对鱼体正常生命活动产生了重大影响,并具有潜在的致癌性．     

苯并[a]芘高效降解菌筛选及其降解特性研究

为获得苯并[a]芘（BaP）污染修复的新菌株,通过梯度提高无机盐培养基（MSM）中BaP浓度的方法,本研究从北京焦化厂污染土壤中筛选出1株能够对液体中高浓度BaP（100 mg/L）降解的霉菌,鉴定结果为可可毛色二孢菌（Lasiodiplodiatheobromae）.并首次对L.theobromae在液体中降解BaP...     

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.     

云南省曲靖燃煤区人群多环芳烃的日暴露特征研究

2007年12月~2008年1月于我国云南省曲靖燃煤区开展人群多环芳烃(PAHs)的暴露评价研究.采用气相色谱/质谱联用(GC/MS)测定了环境样品中16种PAHs的浓度,对暴露的途径及各种PAHs的来源比例进行了分析,以苯并(a)芘(BaP)致癌当量计算了人群日暴露总量.结果表明,人群日饮食暴露量为0.20μg/d,且与WHO报道的水平相当.日空气暴露量为5.26μg/d,占PAHs日暴露总量的96%,表明空气是当地PAHs暴露的主要来源.用BaP当量剂量评价致癌风险时,BaP从数量比3.08%上升至毒性当量比49.6%,低环PAHs毒性当量所占比例明显下降.当地人群BaP平均日暴露量为5.46μg/d,超过了标准限值3μg/d.PAHs暴露给当地环境和人群健康带来了长期危害.     

基于CMAQ模式的中国大气BaP迁移转化模拟研究

在空气质量模式CMAQ中增加了对大气中苯并[a]芘(BaP)传输、转化和沉降过程的模拟,新增了BaP气相/颗粒相间分配模式(K_OA吸收模式)以及BaP化学转换,并采用该模型模拟了2014年1、4、7、10月四个季节代表月中国大气中BaP的浓度以及干湿沉降通量的空间分布特征.模拟结果与部分地区的实测数据进行了初步对比验证,结果显示BaP浓度的模拟值和测量值的量级基本一致,其比值基本在1~2个量级范围内,表明模拟结果处于可接受的范围.基于模型结果,还探讨了BaP浓度与沉降季节变化特征与人为源排放的紧密联系,说明湿沉降是BaP最主要的沉降途径.BaP沉降的空间分布与其排放源和浓度具有密切的联系,并且具有长距离输送的特征等.     

苯并[a]芘对脊尾白虾(Exopalaemon carinicauda)肝脏生物标志物指标的影响

研究了脊尾白虾(Exopalaemon carinicauda)在对苯并[a]芘(benzo[a]pyrene,BaP)的富集(15 d)、释放(15 d)过程中肝脏的生物标志物(EROD、GST、SOD、CAT、GPx、MDA)的响应,结果显示:在富集阶段,第1 d低浓度实验组(0.05 g/L)和高浓度实验组(0.45 g/L)六种生物标志物即均显著受到诱导(P0.05),诱导程度与暴露浓度成正相关,而后不同种类生物标志物呈现差异性的变化趋势,EROD继续上升并在第10 d后达到稳定状态,GST继续上升并在第10 d后受到抑制下降,SOD、CAT、GPx均受到抑制逐渐下降,且均在15 d时低于对照组水平,MDA呈现持续上升的趋势,直接反映了肝脏的氧化损伤程度。在释放阶段,低浓度实验组各生物标志物的恢复速度较高浓度实验组要快,且均能恢复到接近对照组水平(P0.05),高浓度实验组各生物标志物则均未能恢复到对照组水平(P0.05)。实验结果表明,脊尾白虾肝脏具有一定恢复能力,各生物标志物对BaP暴露的敏感性具有一定差异,BaP暴露的浓度及时间是影响各生物标志物响应变化的主要因素。     

镉对黑斑蛙精巢细胞色素P450酶及谷胱甘肽转移酶活性的影响

细胞色素P450酶(CYP450)和谷胱甘肽转移酶(GST)是动物体内主要的解毒酶,在外源毒物的转化和代谢过程中具有重要作用.本文在实验条件下, 将健康性成熟黑斑蛙(Rana nigromaculata)暴露于浓度分别为0.005、0.01、0.05、0.1、0.5和1.0 mg·L^-1的Cd溶液中30 d, 采用荧光分光光度法和紫外分光光度法测定精巢组织乙氧基异酚恶唑脱乙基酶(EROD)、红霉素N-脱甲基酶 (ERND)、氨基比林-N-脱甲基醇酶(APND)和谷胱甘肽转移酶(GST)活性,探讨Cd在精巢的可能代谢过程和毒性机理.结果表明, 与对照组比较, EROD和GST酶活性在0.5和1.0 mg·L^-1 Cd处理组被显著抑制; ERND酶活性在0.01~1.0 mg·L^-1 Cd处理组均被显著抑制;而APND酶活性在各处理组响应变化不明显.结果显示, 精巢EROD、ERND和GST对Cd胁迫的敏感性高于APND, 黑斑蛙精巢中EROD、ERND和GST酶活性的响应可用来评价环境中低水平Cd的污染效应.     

Isolation of marine benzo[a]pyrene-degrading Ochrobactrum sp. BAP5 and proteins characterization

A bacterial strain BAP5 with a relatively high degradation ability of benzo[a]pyrene (BaP) was isolated from marine sediments of Xiamen Western Sea, China and identified as Ochrobactrum sp. according to 16S rRNA gene sequence as well as Biolog microbial identification system. Strain BAP5 could grow in mineral salt medium with 50 mg/L of BaP and degrade about 20% BaP after 30 d of incubation. Ochrobactrum sp. BAP5 was able to utilize other polycyclic aromatic hydrocarbons (PAHs) (such as phenanthrene, pyrene and fluoranthene) as the sole carbon source and energy source, suggesting its potential application in PAHs bioremediation. The profile of total soluble protein from Ochrobactrum sp. BAP5 was also investigated. Some over- and special-expressed proteins of strain BAP5 when incubated with the presence of BaP were detected by two-dimensional polyacrylamide gel electrophoresis, and found to be related with PAHs metabolism, DNA translation, and energy production based on peptide fingerprint analysis through matrix-assisted laser desorption/ionization-time of flight mass spectrometry.     

焦化废水中痕量苯并(a)芘的固相萃取和GC-MS检测方法

基于固相萃取(SPE)和气相色谱/质谱联用技术(GC-MS),建立了一种焦化废水中痕量苯并(a)芘(BaP)的定量分析方法。BaP水样添加甲醇改性后经SPE富集净化、二氯甲烷洗脱、氮吹浓缩、二氯甲烷或乙腈定容后采用GC-MS进行定量分析。结果表明:在优化的样品前处理和分析条件下,BaP标准曲线的线性相关系数R2>0.99,方法回收率为79.6%~85.5%,方法检出限和定量限分别为4.33 ng/L和14.45 ng/L。该方法应用于内蒙古某焦化厂二级生化出水加标样品中BaP的测定,得到BaP的回收率为83.3%~89.3%,表明其灵敏度高、选择性好,适用于焦化废水中痕量BaP的定量检测。     

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.     

Antioxidant responses to benzo[a]pyrene, tributyltin and their mixture in the spleen of Sebasticus marmoratus

It has been reported that there is an interaction between Benzo[a]pyrene （BaP）, a widespread carcinogenic polycyclic aromatic hydrocarbon, and tributyltin （TBT）, an organometal used as an antifouling biocide. This study was therefore designed to examine the potential in vivo influence of BaP, TBT and their mixture on splenic antioxidant defense systems of Sebastiscus marmoratus. The fish were exposed to water containing environmentally relevant concentrations of BaP, TBT and their mixture. Spleens were collected for biochemical analysis after exposure for 7, 25, 50 d and after recovery for 7, 20 d. Cotreatment with BaP and TBT for 7 d potentiated the induction of glutathione peroxidase （GPx） activity by BaP or TBT alone. The cotreatment for 25 and 50 d resulted in inhibition of GPx activity, which was similar to the effect of TBT. Splenic glutathione S-transferase （GST） activities were significantly elevated in S. marmoratus exposed to BaP starting from 7 d and remained high up to 25 d. However, no further activity change was found with prolonged exposure. Cotreatment of BaP and TBT primarily inhibited the GST activity, which was similar to the effect of TBT. Cotreatment with BaP and TBT for 25 or 50 d potentiated the depletion of GSH （glutathione） by BaP or TBT alone. MDA （malondialdehyde） contents in spleen of S. marmoratus were not significantly altered compared with the control during the test period. Spleen, as an immune organ, is sensitive to exposure of BaP or TBT. It should have an effective mechanism to counteract oxidative damage. Antioxidative defense systems in spleen of S. marmoratus should be considered as potential biomarkers. Short-term exposure of BaP or TBT could result in induction of antioxidant defense system. A significant decrease of these indices, such as GSH, GST, GPx might indicate more severe contamination.