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1.
Perch (Perca fluviatilis), were sampled from unpolluted and polluted areas in Swedish coastal waters. The level of aromatic/hydrophobic DNA adducts in liver tissue was analyzed using the nucleas P1 version of the32P-post-labeling assay. The level of total adducts measured in the individual fish from polluted areas was between 6 and 22 nmol of adducts/mol of nucleotides, and in the fish from the reference area between 0.2 and 0.6 nmol of adducts/mol of nucleotides. 相似文献
2.
Turbot (Scophthalmus maximus) and mussel (Mytilus edulis) microsomes were incubated with DNA to examine if microsomal in vitro metabolism of BaP could result in DNA adducts detected by 32P-postlabelling. Turbot DNA was incubated with benzo[a]pyrene (BaP), NADPH and microsomal activating systems prepared from either livers of unexposed turbot, turbot exposed to BaP or beta-naphthoflavone (beta-NF) or digestive glands from mussels. The beta-NF activating system generated the highest levels of DNA adducts detected in this study (451.7 adducts per 10(8) nucleotides) and were distributed in three discrete adduct TLC spots, one of which (97% of the total adducts) co-migrated with the 32P-postlabelled BaP 7,8-diol, 9,10-epoxide-N2-guanine adduct. Fewer adducts (P < 0.05) were generated by BaP-induced microsomes (9.4-30.6 adducts per 108 nucleotides) but levels were higher (P <0.05) than those generated from untreated fish (3.5 adducts per 10(8) nucleotides). Co-incubation with 500 microM alpha-naphthoflavone (alpha-NF) resulted in 97-99% inhibition in adduct formation implicating cytochrome P450-dependent (CYP) bioactivation however there was some evidence for carry over of BaP in the liver microsomal preparations from BaP injected fish. In contrast to the fish activating systems, no DNA adducts were observed when mussel microsomes were incubated with BaP, DNA and NADPH. 相似文献
3.
Gunilla Ericson Gun kerman Birgitta Liewenborg Lennart Balk 《Marine environmental research》1996,42(1-4)
DNA adducts in cod embryos and larvae were analysed by 32P-postlabeling to test the hypothesis that anthropogenic substances, which could form reactive intermediates, are involved in the reproductive failure of cod (Gadus morhua) from the Baltic Sea. A comparison with cod from the Barents Sea was performed. The mean value of DNA adducts in cod embryos/larvae from the Baltic Sea was 2.3 nmol of adducts/mol nucleotides, compared to 0.12 nmol of adducts/mol nucleotides in the embryos/larvae from the Barents Sea. 相似文献
4.
The viviparous blenny (Zoarces viviparus), also known as eelpout, is considered a suitable biomonitor for use in European estuarine and coastal waters. We have previously shown that Z. viviparus from the industrialised Tyne estuary (UK) display certain histopathologies (e.g., ovotestis and hepatic nuclear and cellular pleomorphism), which are associated with contaminant exposure [Mar. Environ. Res. 55 (2003) 137]. Furthermore, the prevalence of these pathologies was higher than in fish collected from a less contaminated reference site (the Alde estuary, UK). Here, tissue samples were collected from Z. viviparus from the Tyne and Alde estuaries (in the spring and autumn) and analysed for DNA adducts using the (32)P-postlabelling assay and for histopathology. Z. viviparus caught from the Tyne during the spring (56.4+/-18.5 adducts per 10(8) nucleotides) and autumn (63.1+/-7.5 adducts per 10(8) nucleotides) exhibited patterns of DNA adducts which indicated exposure to a complex mixture of genotoxins. In contrast, levels of DNA adducts in Z. viviparus from Alde estuary were significantly lower during both sampling periods. Histopathological changes to the gonads included intersex (ovotestis) and germ cell apoptosis (detected via the TUNEL assay) in male fish captured from the Tyne estuary. Further studies on the use of germ cell apoptotic rate at different estuarine sites are required if it is to be used as a reproductive biomarker of contaminant exposure in male fish. 相似文献
5.
Dab (Limanda limanda) were sampled from a number of polluted and unpolluted areas in British coastal waters. The 32P-postlabelling assay was used to analyse the level of aromatic/hydrophobic DNA adducts in pooled samples of liver tissue. The mean levels of DNA adducts detected from areas known to receive anthropogenic pollutants ranged from 4.0 to 26.8 adducts per 10(8) nucleotides, with all sites containing samples displaying DNA adduct profiles consisting of diagonal radioactive zones. In contrast no DNA adducts were detectable in samples from an unpolluted reference site. The ranking of polluted sites based on DNA adduct levels did not correspond with the ranking of sites based on sediment associated polycyclic aromatic hydrocarbon levels, highlighting the problem of linking the presence of contamination with detectable biological responses. No correlation could be found in this study between EROD activity and the level of DNA adducts. 相似文献
6.
Turbot (Scophthalmus maximus) and mussel (Mytilus edulis) microsomes were incubated with DNA to examine if microsomal in vitro metabolism of BaP could result in DNA adducts detected by 32P-postlabelling. Turbot DNA was incubated with benzo[a]pyrene (BaP), NADPH and microsomal activating systems prepared from either livers of unexposed turbot, turbot exposed to BaP or β-naphthoflavone (ß-NF) or digestive glands from mussels. The β-NF activating system generated the highest levels of DNA adducts detected in this study (451.7 adducts per 108 nucleotides) and were distributed in three discrete adduct TLC spots, one of which (97% of the total adducts) co-migrated with the 32P-postlabelled BaP 7,8-diol, 9,10-epoxide-N2-guanine adduct. Fewer adducts (P <0.05) were generated by BaP-induced microsomes (9.4–30.6 adducts per 108 nucleotides) but levels were higher (P <0.05) than those generated from untreated fish (3.5 adducts per 108 nucleotides). Co-incubation with 500 μM α-naphthoflavone (α-NF) resulted in 97–99% inhibition in adduct formation implicating cytochrome P450-dependent (CYP) bioactivation however there was some evidence for carry over of BaP in the liver microsomal preparations from BaP injected fish. In contrast to the fish activating systems, no DNA adducts were observed when mussel microsomes were incubated with BaP, DNA and NADPH. 相似文献
7.
DNA adducts in gills and digestive gland, as well as polycyclic aromatic hydrocarbon (PAH) tissue levels were analysed in blue mussels (Mytilus spp.) from Nordic coastal areas (Iceland, Norway and Sweden) with diffuse or point sources of PAHs of various origins. Both DNA adduct and PAH tissue levels were generally low, indicating low PAH exposure to the mussels in the areas studied. DNA adducts were found to be higher in gills than in digestive gland of the mussels at all sites studied. Elevated DNA adduct levels in gills were found at 6 sites out of 18 compared to reference sites in respective coastal zones. Adduct levels ranged from 0.5 to 10 nmol adducts/mol normal nucleotides, being highest in mussels from Reykjavík harbour, Iceland (intertidal mussels), and from Fiskaatangen, Norway (subtidal mussels). Total PAH tissue levels in the mussels ranged between 40 and 11,670 ng/g dry wt., and were significantly correlated with DNA adduct levels (r(2)=0.73, p<0.001). PAH ratio values indicated that the PAHs were in most cases of pyrolytic origin, but with petrogenic input near harbours and an oil refinery. 相似文献
8.
Telli-Karakoç F Ruddock PJ Bird DJ Hewer A Van Schanke A Phillips DH Peters LD 《Marine environmental research》2002,54(3-5):511-515
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. 相似文献
9.
Copper accumulation, and its effect on growth and hematological parameters were investigated in juvenile rockfish, Sebastes schlegeli (mean length 11.83+/-0.03 cm, and mean weight 26.02+/-0.23 g), after sub-chronic dietary Cu exposure (0, 50, 125, 250 and 500 mg/kg) for 60 days. The profile of Cu accumulation among tissues in rockfish is dependent on the exposure periods and Cu concentration. Liver of rockfish is a more important storage tissue than other tissues, and the order of Cu accumulation in tissues was liver > intestine > kidney > gill > muscle. Cu reduced the growth rate, and there was an inverse relationship between growth and Cu concentration (> 50 mg/kg). The RNA:DNA ratios were not affected by exposure and there was no correlation between growth rate and RNA:DNA ratio in the liver and muscle. There was no significant effect of exposure on blood parameters except for magnesium. Cu exposure increased GOT and GPT serum concentrations with increasing time and dose. 相似文献
10.
应用荧光定量PCR技术, 检测了TLR21 基因在牙鲆(Paralichthys olivaceus)感染迟缓爱德华 氏菌(Edwardsiella tarda)后, 在0 h、1 h、3 h、6 h、12 h、1 d、3 d、6 d 后, 在心脏、肝脏、脾脏、 头肾、鳃、小肠、肌肉和血的时空表达特征, 并探讨了它们与牙鲆先天免疫反应的关系。结果表明, 大 多组织在感染病原6 h 后TLR21 基因表达明显上调, 尤其是头肾和小肠。头肾6 h 的表达量达到了 对照组的59.3 倍, 小肠6 h 的表达量为对照组的38.6 倍。迟缓爱德华氏菌感染引起牙鲆体内各组织 中TLR21 的上调表达和变化, 为研究牙鲆对迟缓爱德华氏菌的防御机制提供了理论依据。 相似文献
11.
The chronic effects of polycyclic aromatic hydrocarbons (PAHs) on ovary development, total hepatic lipids and plasma sex- and corticosteroid levels in female flounder (Platichthys flesus) were examined. Sexually mature feral female flounder were exposed via the diet to phenanthrene (0.5, 2.5 or 12.5 nmol/g food) or chrysene (0.4 nmol/g food) for 12 weeks, during the previtellogenic phase of the annual reproductive cycle. PAH exposure did not directly affect germ cell development since no structural and/or developmental differences were observed between control and exposed fish. On the contrary, all treatments resulted in altered plasma steroid levels. The most pronounced effect was the significant decrease in plasma 17 beta-estradiol to 19 +/- 11%, 27 +/- 7%, 63 +/- 20% and 61 +/- 12% in relation to control fish, respectively, in flounders exposed to 12.5, 2.5 or 0.5 nmol phenanthrene/g food and 0.4 nmol chrysene/g food. Impaired ovarian growth was not observed, most likely because experiments were ended before the period of vitellogenesis, even though a non-significant general decline in total hepatic lipids could be observed. Moreover, all exposed flounders, except fish fed with the highest amount of phenanthrene, showed a negative correlation between plasma 17 beta-estradiol and 17 alpha-hydroxyprogesterone levels (r = -0.46). One possible explanation is that PAH action may be mediated by a specific inhibition of steroidogenic enzymes. These findings provide evidence that selected PAHs are antiestrogenic xenobiotics with the capability to impair female teleost reproductive function. 相似文献
12.
抗细胞凋亡因子(Defender against cell death 1,DAD1)是一种重要的内源性细胞凋亡抑制因子.采用RACE-PCR方法,结合SMART cDNA合成方法、抑制性差减杂交技术,从青石斑鱼脾脏中克隆DAD1并研究其在正常的青石斑鱼和注射了脂多糖的青石斑鱼各组织中的表达模式.结果表明:DAD1基因cDNA全长721 bp,包含一个342 bp的完整阅读框,编码一个由113个氨基酸组成的蛋白.RT-PCR半定量分析显示DAD1在正常的青石斑鱼和注射了脂多糖的青石斑鱼的脾、心、头肾、肾、肝中都有转录,在脾、肝中表达水平较高,而在肌肉中未见明显表达.DAD1在注射了脂多糖的青石斑鱼各组织中的表达量均高于正常的青石斑鱼. 相似文献
13.
采用组织学方法,获得患肾肿大症美国红鱼的心、脾、肾、肝、肠和鳃6种组织,利用组织切片以及超薄切片电镜技术对患病组织的病理变化进行了研究分析,探讨该病的致病机制。患病红鱼解剖后主要症状为:体表溃烂,部分病鱼鳍条上有肿块;病鱼肾脏显著肿大,体积甚至达到正常肾脏的1()倍;肝脏、脾脏及胆囊等也有肿大现象;有些病鱼脏器上有白色结节。组织病理显示:美国红鱼肾肿大症的主要表现为肾脏有典型的肉芽肿,大量炎性细胞浸润,肾小管管壁破裂、坏死;肝脏中肝实质有充血现象,实质细胞呈空泡变性;脾脏充血,现大量病变的红细胞,有脂肪样变。超徽病理显示:肾近曲小管上皮细胞凋亡现象严重,肾小管内刷状缘排列稀疏;肝细胞坏死严重,肝细胞界限模糊不清;脾脏组织脾窦扩张,有大量凋亡红细胞和淋巴细胞。实验结果表明:美国红鱼肾肿大症的组织细胞结构发生明显的病理变化,肾脏和肝脏等主要组织器官受到严重损伤病变,使鱼体不能进行正常的生命活动,最后导致死亡。 相似文献
14.
The effects of waterborne nitrite (3 mg/l NO2) on channel catfish were studied to evaluate changes in hematological parameters and phase I-II biotransformation in liver slices. Nitrite-exposed fish had significantly higher methemoglobin, blood and liver nitrite, and significantly lower pO2 than control fish. Total phase I-mediated metabolism of 7-ethoxycoumarin (EC) was not altered in nitrite-exposed fish compared with control fish (291 +/- 43 and 312 +/- 20 pmol/mg/h, respectively). However, phase II glucuronosyltransferase-mediated metabolism of 7-hydroxycoumarin (HC), both as a phase I metabolite of EC and as a parent substrate, was elevated in nitrite-exposed fish (204 +/- 17 and 1007 +/- 103 pmol/mg/h, respectively) as compared to control fish (149 +/- 14 and 735 +/- 87 pmol/mg/h) (P < 0.05). Sulfotransferase-mediated metabolism of HC (as a metabolite of EC and as a parent substrate) was not notably altered in nitrite-exposed fish (95 +/- 16 and 617 +/- 33 pmol/mg protein/h, respectively) as compared with control fish (118 +/- 24 and 575 +/- 55 pmol/mg/h, respectively). These studies indicate that in vivo nitrite exposure and associated changes in hematological parameters do not appear to affect hepatic phase I EC biotransformation in channel catfish. However, subtle but significant changes in phase II glucuronidation, but not sulfation activity, were observed. The mechanism of these alterations is unclear. However, the data suggest that environmentally realistic concentrations of nitrite may affect the dynamics of conjugative metabolism in exposed fish. 相似文献
15.
16.
Conjugation of phenolic xenobiotics and metabolites through sulfation and glucuronidation is an important biotransformation pathway. Sulfotransferases (SULT) are generally considered non-inducible, while some UDP-glucuronosyltransferase (UGT) isoenzymes are co-induced with cytochrome P450-1A by Ah-receptor ligands. To test these assumptions for two fish species, we measured sulfation and glucuronidation of 9-hydroxybenzo[a]pyrene in 3-methylcholanthrene (3-MC) treated channel catfish (Ictalurus punctatus) and in mummichog (Fundulus heteroclitus) from the creosote contaminated Atlantic Wood site in the Elizabeth River, VA. The results show a significant induction of both UGT and SULT activity in 3-MC treated catfish, linked to the expected induction of EROD activity. In mummichog, significant induction of UGT was measured at the contaminated site over the reference site (King's Creek, VA), as well as extremely low SULT activities at both sites. Western blots, using a polyclonal antibody for catfish phenol-type SULT, confirmed the absence of phenol-type SULT in mummichog. Residual, though slightly inducible, SULT activity may be attributed to other SULT isoforms. 相似文献
17.
B. A. McKeown 《Marine environmental research》1978,1(2):119-123
Chlorine-labelled Bunker C oil was used to measure the differential accumulation in various fish tissues between a hydrocarbon and a hydrocarbon/oil dispersant mixture. There is an increased movement of the emulsified oil across the gill structure although accumulation by this tissue is similar for both test conditions. The liver and kidney showed significantly higher levels of the oil/dispersant mixture whereas muscle accumulations were less dramatic. The amounts of Bunker C found in the gills, liver and kidney were considerably higher than that found in the muscle. Consideration was given to the varying capability of the blood to carry polar, compared with non-polar, compounds. 相似文献
18.
Diagnosis of iridovirus in large yellow croaker by PCR 总被引：1，自引：0，他引：1
A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus(LYCIV) is described,which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen.Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus(RSIV) and sea bass iridovirus(SBIV),suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone.The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes,the expected fragment was detected from spleen DNA samples of infected fishes,whereas no fragments were amplified from healthy fish spleen DNA,white spot syndrome baculoviruses(WSBV) DNA and pseudorabies virus(PRV) DNA.Detection limit of this method was 10-7 ng positive plasmid DNA containing target sequence,equal to about 100 virions.In the infected experiment,first positive detection(1/4) appeared at Day 3 post-infection,all fish(4/4) tested positive at Day 7,however obvious symptoms were observed at Day 8,so LYCIV infection could be detected prior to the appearance of obvious symptoms.These results indicate that this PCR method could be used for early,rapid and specific detection of LYCIV infection. 相似文献
19.
Haasch ML 《Marine environmental research》2002,54(3-5):297-301
The mummichog (Fundulus heteroclitus) has been shown to be responsive to peroxisome proliferating agents (PPAs). Peroxisomes function as important sites for fatty acid beta-oxidation. Peroxisome proliferation by PPAs or starvation can lead to changes in the size and number of peroxisomes and the expression of omega-hydroxylases (CYP2K1/2M1 in rainbow trout). Mummichogs were subjected to 96 h fasting or 96 h recovery from fasting. Expression of PMP70- and CYP2K1/2M1-like proteins in vehicle-treated or non-treated controls was compared in both males and females. Fasting and vehicle produced decreases in PMP70- and CYP2K1/2M1-like proteins in both males and females. In reproductive females, decreases due to fasting and vehicle treatment were greater than in female fish that were not gravid. Recovery from fasting resulted in levels of CYP2K1/2M1 near control levels in males while in recovered females, about 2-fold higher levels compared to controls were noted. These results indicate that gender, reproductive status and diet can produce changes in the expressed levels of peroxisomal PMP70 and microsomal CYP2K1/2M1-like proteins in the mummichog. 相似文献
20.
Benzo[a]pyrene (BaP), a procarcinogenic polycyclic aromatic hydrocarbon (PAH), is bioactivated to BaP diol-epoxides (BPDEs) that can form adducts with DNA and blood proteins. We report here for the first time the in vivo formation of adducts between BPDE and plasma albumin (Alb) from two fish species experimentally exposed to BaP. Brook trout (Salvelinus fontinalis) received either a single i.p. dose (10 mg/kg) or two separate i.p. doses (25 mg/kg; 7 days apart) of BaP, and blood was collected 2 (single exposure) or 3 (multiple exposure) days post-treatment. Arctic charr (Salvelinus alpinus) received 10 i.p. doses (3 mg/kg; a single dose every 6 days), and blood was collected 2 days after the second, sixth, and 10th injections. BPDE-Alb adducts were measured by an improved HPLC/fluorescence method developed to detect and quantify BaP-tetrols released after acid hydrolysis of adducted Alb. HPLC/fluorescence chromatograms of Alb from BaP-treated fish revealed only BaP-tetrol I-1, thus indicating the formation of adducts exclusively via the (+)-anti-BPDE metabolite. Levels of (+)-anti-BPDE-Alb adduct ranged from 0.68 to 19.6 ng of tetrol I-1 per gram of Alb. Notably, adduct level was not related to BaP dose and there was no accumulation of adducts with repeated exposure, which may indicate a very short half-life (< 2 days) of plasma Alb in fish. The data suggest that BPDE-Alb adducts in fish could be useful as a non-destructive biomarker of recent exposure to bioactivated BaP. 相似文献