苯并[a]芘对雄性褐菖鲉精巢成熟的影响

本实验采用雄性褐菖鲉作为实验对象,腹腔注射0.5×10-6、1×10-6、5×10-6、10×10-6(m/m)苯并[a]芘,7d后苯并[a]芘处理组精巢睾酮水平和睾酮与17β-雌二醇的比值都有上升,但都未表现出显著性差异(P>0.05).腹腔注射50×10-6(m/m)苯并[a]芘,11d后取出精巢,常规石蜡切片,结果显示雄性褐菖鲉的精巢,精小叶腔和输出管中精子密集程度明显增加,次级精母细胞减少,精巢壁厚度变薄.结果提示,苯并[a]芘对褐菖鲉具有类雄激素效应.     

蒽与苯并[a]芘对海洋微藻致毒影响的比较研究

新月菱形藻 ,球等鞭金藻 870 1和亚心形扁藻 ,分别用不同浓度的蒽和苯并 [a]芘胁迫72 h,测定三种藻的生长速率 ,叶绿素 a含量。实验表明 ,三种微藻对蒽的敏感性由高到低依次为新月菱形藻、球等鞭金藻 870 1和亚心形扁藻 ,它们 72 h的半抑制浓度 EC50 分别是 0 .0 6 0 mg/L、0 .0 6 5mg/L和 0 .0 94 mg/L。而对苯并 [a]芘的敏感性由高到低依次为亚心形扁藻、新月菱形藻和球等鞭金藻 870 1,它们的 72 h· EC50 分别是 0 .0 76 mg/L、0 .0 88mg/L 和 >0 .10 mg/L(在本实验范围内未测到 )。叶绿素 a含量与各自的相对增长率呈一定的正相关。不同的海洋微藻对蒽和苯并 [a]芘的敏感性不同。     

苯并(a)芘和芘对梭鱼肝脏DNA损伤的研究

用苯并(a)芘、芘以及它们的等量混和物,分别在浓度为0.1,1,10,20,50μg/dm3浓度下对梭鱼暴污,5d后取梭鱼肝脏和鳃用碱解旋法分别测定其DNA的损伤,结果随着污染物浓度的增加,肝脏DNA损伤程度增加;在相同浓度下,苯并(a)芘和芘的联合毒性大于苯并(a)芘和芘分别作用时的毒性之和。所以苯并(a)芘和芘对DNA损伤的联合作用应为加强作用。     

苯并[a]芘(BaP)对褐菖鲉(Sebasticus marmoratus)肝CYP1A1酶活性、基因表达及蛋白表达的影响

为了了解苯并[a]芘(BaP)对鱼类细胞色素P4501A1(CYP1A1)表达的影响,以褐菖鲉(Sebasticus marmoratus)为实验材料,采用体内实验,研究其在经过不同浓度(0.1、1、10、20、50mg/kg鱼体重量)的BaP诱导后,鱼体肝脏研究CYP1A1基因表达的情况,筛选出后续时间-效应实验中BaP注射的最佳浓度,研究BaP诱导6h、12h、1d、3d、7d后(质量浓度为20mg/kg鱼体重量)鱼体肝脏CYP1A1酶活性、基因表达和蛋白表达的情况。结果表明:剂量-效应实验中,20mg/kg鱼体重量为最佳浓度,此浓度下,基因表达在各组中变化最显著。时间-效应实验中,较空白对照组而言,染毒6h、12h和1d后,EROD酶活性显著增加。3d后开始下降,与对照组相比变化不大,7d后酶活性又发生上调。半定量RT-PCR结果表明,各染毒组与对照组相比,CYP1A1基因表达量都发生了上调,呈现先上升后下降的趋势。其中,6h和12h组相对表达量极显著增加,1d后开始下降且与3d和7d组相比变化不明显。Western blot结果表明,蛋白表达量在染毒12h后表现出显著的诱导效应,随着时间的延长略有回落,但与对照组相比仍有显著性差异。研究表明:BaP对褐菖鲉CYP1A1具有较强的诱导作用。一定质量浓度的BaP注射于褐菖鲉不同的时间后,能诱导褐菖鲉活体EROD酶活性、CYP1A1基因m RNA表达及蛋白表达,并随着时间的延长呈现先诱导后抑制的趋势。这说明BaP作为诱导剂对CYP1A1酶活性和蛋白表达的作用机制可能与调控CYP1A1的转录水平有关。     

苯并（a）芘对大弹涂鱼肝脏超氧化物歧化酶活性的影响

在实验生态条件下，研究了在苯并（a)芘（BaP)胁迫下大弹涂鱼肝脏超氧化物歧化酶（SOD）活性的变化，结果显示：暴露3d时，不同BaP含量组大弹涂鱼肝脏SOD活性无显著差异（P＞0．05），而暴露7d时，随着B aP含量的升高，0.5mg/dm^3 BaP含量组SOD活性被显著诱导（P＜0．05），为对照组的1．87倍；随着暴露时间的延长，各含量组肝脏SOD的活性均表现出不同程度的下降趋势，其中对照组肝脏SOD活性显著降低，表明SOD活性易受污染以外的因素，如水体容量、铒料、光照等的影响。污染解除后，0.5mg/dm^3含量组SOD活性显著升高，表明大弹涂鱼肝脏仍具有较强的生理调节机能，也可能表明SOD活性的变化相对于环境因子改变的“迟滞性”。以上这些结果表明SOD有可能作为大弹涂鱼受BaP胁迫的生物指标。     

苯并(a)芘对紫菜(Porphyra)的致突变效应

苯并(a)芘(Benzo(a)pyrene)是石油组份中的一种重要的具致癌作用的多环芳香烃化合物。由于燃烧和石油污染等多种原因,在大气、水体和海洋中都能检出它的存在,特别是在海洋中,这种化合物有的沉积于沉积物中,有的被生物体吸收,有的附着在生物体上,有的存在于水体中,因而构成了海洋中一类特殊的污染物。特别引起科学家注意的是这物种质可为某些海洋生物所富集或贮存,并经食物链而最后进入人体。     

苯并[a]芘在脊尾白虾（Exopalaemon carinicauda）体内富集的动力学研究

通过室内实验模拟了脊尾白虾（Exopalaemon carinicauda）对苯并[a]芘（benzo[a]pyrene,Ba P）的生物富集,通过非线性曲线拟合获得脊尾白虾对Ba P的富集动力学参数（吸收速率常数k1、释放速率常数k2、生物富集因子BCF、平衡状态下脊尾白虾体内Ba P含量CAmax、生物学半衰期B1/2）.各动力学参数分别为：k1范围为12.34～24.96,平均值为18.80,k2范围为0.06～0.10,平均值为0.08,BCF范围为208.41～248.03,平均值为228.02、CAmax范围为12.40～93.79 ng/g,平均值为46.78 ng/g,B1/2范围为6.89～11.71 d,平均为8.95 d.脊尾白虾对Ba P的吸收速率常数k1、释放速率常数k2、BCF均随外部水体中Ba P浓度的增大而减少,CAmax、B1/2随外部水体中Ba P浓度的增大而增大.     

苯并(a)芘、芘及其混合物暴露对梭鱼肝脏谷胱甘肽硫转移酶活性的影响

在实验生态条件下,观察苯并(a)芘、芘及其等浓度混合物暴露对梭鱼(Mugil so-iuy)肝脏谷胱甘肽硫转移酶(GST)活性的影响。结果显示,在7d的暴露中,苯并(a)芘、芘对肝脏GST活性的影响主要为诱导效应,芘对GST活性的诱导比苯并(a)芘强。混合物在15d的暴露中未观察到GST活性的诱导,而是在暴露的后期出现GST活性的抑制。实验表明,肝脏GST活性的诱导指示受到PAHs污染胁迫,而GST活性抑制则是受到较长时间或较严重的污染。     

苯并（a）芘诱导下岩虫CYP基因的表达特征

Rapid amplification of cDNA ends(RACE) and real-time polymerase chain reaction(RT-PCR) were carried out to analyze the CYP4 gene expression in polychaete Marphysa sanguinea exposed to benzo[a]pyrene(BaP) in this study. The full length of MsCYP4 cDNA was 2 470 bp, and it encoded 512 amino acids. The deduced amino acid sequence showed 47% identity with CYP4 F from frog Xenopus tropicalis and shared high homology with other known CYP4 sequences. To analyse the role of CYP4 in protecting M. sanguinea from BaP exposure, three BaP groups were established: 0.5, 5 and 50 μg/L. Polychaetes were sampled after 3, 7 and 12 d. At 0.5 μg/L, the effect of BaP on MsCYP4 gene expression increased with time prolonged. MsCYP4 gene expression curve showed Ushaped trend with time in 5 and 50 μg/L BaP groups. Therefore, MsCYP4 gene may play an important role in maintaining the balance of cellular metabolism and protecting M. sanguinea from BaP toxicity.     

Phototoxicity of pyrene and benzo[a]pyrene to embryo-larval stages of the Pacific oyster Crassostrea gigas

There is a growing body of evidence to suggest that certain polycyclic aromatic hydrocarbons (PAHs) pose a greater hazard to aquatic organisms than previously demonstrated, due to their potential to cause photo-induced toxicity when exposed to ultraviolet (UV) radiation. The consequences of photo-induced toxicity are reported here for embryo-larval stages of the pacific oyster Crassostrea gigas, following exposure to pyrene and benzo[a]pyrene. During laboratory investigations, significant increases in toxicity were observed in the presence of environmentally attainable levels of UV-radiation, compared with embryos exposed to PAH alone, at levels previously deemed to have little acute biological effect. The phototoxicity of pyrene and benzo[a]pyrene completely inhibited the development to the D-shell larval stage when embryos were simultaneously exposed to 5 microg l(-1) PAH and ultraviolet light (UVB = 6.3 +/- 0.1 microW/cm2 and UVA = 456.2 +/- 55 microW/cm2). A linear relationship was also demonstrated for benzo[a]pyrene phototoxicity with decreasing UV light intensity.     

Trophic transfer of benzo[a]pyrene metabolites between benthic marine organisms

This study was undertaken to determine the potential for trophic transfer of polycyclic aromatic hydrocarbon (PAH) metabolites from infaunal organisms to bottom-feeding fish. Winter flounder, Pseudopleuronectes americanus, were given single oral doses of ground polychaetes (Nereis virens), either treated with pure [¹⁴C]henzo[a]pyrene (BaP) or containing a mixture of naturally produced radiolabeled BaP and BaP metabolites. Fish were sacrificed 24 h after feeding and total accumulated radioactivity and metabolite class profiles determined in major organs. Metabolites produced by worms were absorbed by flounder, although as a percentage of dose given they were less available than parent BaP. Comparison of metabolite profiles in the worm diet and in target organs in the fish indicated that metabolites accumulated through the diet can be further modified by the prey organism and can lead to the formation of bound residues. These results demonstrate that PAH metabolites in the diet are available for accumulation. Furthermore, metabolites absorbed appear to be susceptible to metabolic alteration by consumer organisms.     

Effects of benzo[a]pyrene on DNA damage and histological alterations in gonad of scallop Chlamys farreri

To investigate the biological impact of polycyclic aromatic hydrocarbons (PAHs) on reproductive system, scallops Chlamys farreri were continuously exposed to benzo[a]pyrene (BaP) (0.5, 3, 10 μg L^?1) during 15 days. DNA damage and histological alterations were examined in female gonad. DNA strand break levels significantly increased after 12 h exposure, and remained high and significantly different from control values until the end of the exposure. In the ovaries of the scallops exposed to 10 μg L^?1 BaP for ten days, histological analysis showed that the cytoplasts of the oocytes in the outer layer of the ovaries became sparse, and the nuclear membranes were obscure. Damaging effects on ovaries and oocytes were more severe after 15 days exposure. Degenerating oocytes increased and the connective tissue of the ovary envelops became loose. Electron microscopic examination revealed ultrastructural aspects of degenerating oocyte and degenerated oocyte after 15 days exposure.     

Interactive effects of cadmium and benzo[a]pyrene on metallothionein induction in mummichog (Fundulus heteroclitus).

Previous experiments demonstrated that exposure of mummichog to cadmium (Cd) in combination with benzo[a]pyrene (BaP) caused a higher mortality than would be expected from simple additive effects. Experiments are described here that investigated whether BaP exposure inhibits the induction of metallothionein (MT), a major detoxifying protein for Cd, or if reactive BaP metabolites compete with Cd for binding sites on MT. Fish were injected with or without BaP (18 mg/kg) in combination with a low (1 mg/kg) or high (3.2 mg/kg) dose of Cd, and in one treatment BP was dosed 4 days after Cd. The results showed a rapid induction of MT to 1.5 mg/g wet weight liver, 1 day after injecting the low Cd dose. Simultaneous BaP exposure significantly delayed the induction of MT, for both low and high Cd doses, and BaP temporarily lowered the induced MT concentration when dosed 4 days after induction by Cd. To test if binding of BaP metabolites to MT reduces the detoxification potential for Cd, microsomes of CYP1A-induced fish were incubated with MT and radiolabeled BaP. Active metabolism of BaP was observed by high-performance liquid chromatography analysis, but no association of BaP metabolites with MT was found. Neither could this be demonstrated in vivo, in liver MT isolated from mummichog dosed with 3H-BaP and Cd. These results suggest that increased toxicity of Cd in combination with BaP exposure is likely to be caused by inhibited MT synthesis, rather than by interference of BaP metabolites with Cd binding on MT.     

In-vitro metabolism of benzo[a]pyrene and benzo[a]pyrene-7,8-dihydrodiol by liver and intestinal mucosa homogenates from the winter flounder (Pseudopleuronectes americanus)

Freshly prepared homogenates were used to assess the relative ability of winter flounder (Pseudopleuronectes americanus) liver and intestinal mucosal cells to metabolize the polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (BaP) and its proximate carcinogenic metabolite, BaP-7,8-dihydrodiol (7,8-Diol). Data obtained from homogenates prepared from fish previously fed β-naphthoflavone (BNF) indicated that both tissues had similar abilities to metabolize either BaP or 7,8-Diol on a per gram of protein basis. Metabolite profiles produced indicate that water-soluble metabolite formation is favored at low doses. These findings support the hypothesis that the intestine plays an important role in first-pass metabolism of dietary carcinogens in the winter flounder.     

Correlative changes in metabolism and DNA damage in turbot (Scophthalmus maximus) exposed to benzo[a]pyrene

Juvenile turbot (Scophthalmus maximus) were injected intraperitoneally with either corn oil or 5 mg/kg benzo[a]pyrene (BaP) dissolved in corn oil and sampled I and 3 days after injection. After 1 day, no elevation of 7-ethoxyresorufin O-deethylase (EROD) activity was observed, however bile metabolites (BaP-7,8 dihydrodiol representing 70% of the total metabolites) and a single hepatic DNA adduct spot (0.47 adducts/10(8) nucleotides) identified by 32P-postlabelling were formed. No BaP metabolites or DNA adducts were observed in either control or carrier control fish. Fish sampled after 3 days reported 5-fold higher (P < 0.05) levels of EROD activity, a shift in the bile metabolite profile towards BaP phenol formation (1OH and 30H BaP comprising up to 60% of total metabolites detected) and the formation of two adduct spots (0.86 and 0.71 adducts/10(8) nucleotides). These results show that BaP can be metabolised and form hydrophobic DNA adducts in turbot without EROD elevation. Following EROD elevation, a shift in the profile of both BaP metabolites and BaP metabolite-DNA interactions occurs indicative of other oxidative processes.     

Evidence for resistance to benzo[a]pyrene and 3,4,3'4'-tetrachlorobiphenyl in a chronically polluted Fundulus heteroclitus population

Halogenated aromatic hydrocarbons (HAHs) and polynuclear aromatic hydrocarbons (PAHs) are major environmental contaminants. Fish species that are chronically exposed to these compounds can develop resistance to their toxic effects. In all fish species studied to date, toxicant resistance has been accompanied by decreased inducibility of the xenobiotic metabolizing enzyme, cytochrome P450 1A (CYPIA). CYP1A induction is mediated through the Aryl Hydrocarbon Receptor (AHR). Although these compounds mediate their effects through this pathway, there have been resistant populations in which one chemical class cannot induce CYPIA expression (HAHs) while the other (PAHs) can. Resistance to PAHs was examined in a HAH-resistant population of Fundulus heteroclitus collected from a site contaminated with both compound classes (Newark Bay, NJ). Fish were injected intraperitoneally with the HAH 3,4,3',4'-tetrachlorobiphenyl (PCB77), benzo[a]pyrene (B[a]P, a PAH) or vehicle and sacrificed after 2 (B[a]P) or 5 days (PCB77, vehicle). We found no significant increase in CYP1A mRNA levels in resistant Newark Bay F. heteroclitus treated with either B[a]P or PCB77, while there was a 3.9 fold (PCB77) and 4.2 fold (B[a]P) increase in CYP1A mRNA in Flax fish relative to controls. AHR labeling studies revealed significantly (P < 0.05) lower levels of hepatic AHR in Newark fish (1,770 +/- 1,693.2 DPM) relative to Flax fish (6,082.5 +/- 1,709.9 DPM). Overall, these data suggest Newark F. heteroclitus are resistant to both PAHs and HAHs at the level of CYP1A mRNA, which might be mediated, in part, though lower expression of AHR. We are currently studying the promoter sequence to determine its role in chemical resistance.     

A survey of in vivo benzo[alpha]pyrene metabolism in small benthic marine invertebrates.

A micro-extraction technique was used to examine in vivo polycyclic aromatic hydrocarbon (PAH) metabolism in 10 small invertebrate species exposed to sediments amended with 3H-benzo[alpha]pyrene (BaP). Phyla examined included Mollusca (Hydrobia totteni, Ilyanassa obsoleta, Yoldia limatula, and Gemma gemma), Annelida (Nereis succinea, Pectinaria gouldii, Haploscolopolous sp., and Capitella sp. 1) and Arthropoda (Edotea triloba, and Gammarus mucronatus). Organisms were exposed to BaP-labeled sediments, harvested, and parent BaP separated from all polar metabolites by liquid extraction The percent of BaP-derived radio-activity present as polar metabolites ranged from 96% for N. succinea to 7% for P. gouldii. Wide ranges in metabolic capability were also observed between species in the other two phyla examined. Reverse-phase HPLC analysis of extracts of representative species from each phyla indicated that all these organisms form bay region metabolites, with two species forming the 7,8-dihydrodiol (N. succinea and G. mucronatus). In light of the high variability in metabolic capability observed within each phylum, species-specific information on metabolic ability should be obtained before assessing bioaccumulation, critical body burdens, or trophic transfer of PAHs in invertebrates.