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1.
Large AT Shaw JP Peters LD McIntosh AD Webster L Mally A Chipman JK 《Marine environmental research》2002,54(3-5):493-497
Levels of polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene (B[a]P) were at least seven-fold higher in mussels sampled from a polluted site (Loch Leven, in Scotland, UK) compared to a nearby clean reference site (Loch Etive) throughout the year 2000. Levels of DNA strand breaks (alkaline COMET assay) using both gill and digestive gland nuclei were similar at both sites despite the difference in contaminant load (total PAH). In contrast, mussels collected from a reference site (Port Quin, Cornwall, UK) had an increase in DNA strand breaks in digestive gland cells following laboratory exposure to B[a]P-dosed Isochrysis galbana. However, after 14 days high dose (20 ppb-exposed diet) animals had returned to levels similar to the controls. There was no evidence of increased necrosis or apoptosis after treatments. The results from these two studies suggest that an adaptive response may prevent ongoing DNA damage in mussels exposed to high levels of B[a]P and PAH contamination. 相似文献
2.
Nigro M Frenzilli G Scarcelli V Gorbi S Regoli F 《Marine environmental research》2002,54(3-5):517-520
The ability of benzo[a]pyrene, Aroclor 1254, 2-3-7-8-tetrachlorodibenzo-p-dioxin and beta-naphthoflavone to induce DNA strand breaks (SB) and apoptosis in erythrocytes of the European eel (Anguilla anguilla) was investigated following by in vivo exposure. DNA damage was evaluated by the Comet assay, while the diffusion assay was used to investigate the induction of apoptosis 7 days after a single intraperitoneal administration. 2-3-7-8-Tetrachlorodibenzo-p-dioxin induced the highest genotoxic effect, followed by benzo[a]pyrene, while the other two substances had limited effects. A significant induction of apoptosis was observed at the highest doses after exposure to benzo[a]pyrene, when DNA damage was also elevated. The occurrence of apoptotic cells after exposure to Aroclor, 2-3-7-8-tetrachlorodibenzo-p-dioxin and beta-naphthoflavone was quite variable and did not show clear dose-related responses. The role of oxidative stress in mediating DNA damage was also discussed. 相似文献
3.
We examined the link between DNA strand breaks and hatching rates in grass shrimp, (Paleomonetes pugio), embryos exposed to 0.2 microM benzo[alpha] pyrene (BP), 5 microM cadmium (Cd) and 330 kJ/m(2) UV light, either alone or together. After exposure, embryos were transferred to clean seawater with or without 5 microM Cd. Hatching rates and DNA strand breaks (Comet Assay) were determined. DNA lesions caused by exposure to BP, UV light, or BP/cadmium were rapidly repaired and were not associated with any effects on hatching. Exposure to Cd after exposure to BP or UV did not affect embryological development or DNA repair. Exposure to BP/UV resulted in a high level of DNA lesions which were slowly repaired. Exposure to cadmium following BP/UV exposure inhibited hatching and DNA repair. Adducts formed during exposure to BP/UV exposure may be difficult to excise or may saturate the nucleotide excision repair system. 相似文献
4.
Micronucleus (MN) frequency is generally accepted as a marker of chromosomal damage and has been studied in a variety of cells and species. In previous work, we detected significant dose-related MN increases in the epithelial-like gill cells and agranular haemocytes of Mytilus galloprovincialis treated with benzo[a]pyrene, a well-known mutagenic pollutant. In addition, we have studied micronuclei and other nuclear abnormalities in mussels collected from the Venice lagoon (Italy). Frequency changes, possibly related to genotoxic/toxic stress, in both granular and micronucleated cells from gills and haemolymph, were detected. Environmental data suggest the effect of genotoxic pollutants and the importance of cell replication in the interpretation of micronucleus frequencies. 相似文献
5.
Contribution of apoptosis to observed dna damage in mussel cells 总被引：1，自引：0，他引：1
The comet assay employs the expression of DNA strand breaks by alkaline treatment to measure DNA damage. In contrast to other similar alkaline treatment assays the comet assay incorporates the microscopic examination of damage to individual cell nuclei. Because individual nuclei are examined, features unique to apoptotic cells can be identified. Apoptosis, programmed cell death, is characteristically an orderly sequence of events that ultimately leads to the complete disassembly of a cell. One consequence of apoptosis is the induction of endogenous nucleases which results in the fragmentation of DNA. Apoptosis can be triggered by a variety of stimuli, giving the misleading impression of genotoxic damage if this cytotoxic mechanism of DNA fragmentation is not taken into consideration. In this study mussel cells were exposed to a variety of toxicants and the cytotoxic (apoptotic) contribution to observed DNA damage was determined. A number of stressors induced apoptosis quite rapidly, within hours of exposure. The comet assay detected apoptosis and under certain circumstances apoptosis was found to be the major contributor to observed DNA damage. 相似文献
6.
Biological and procedural factors can influence DNA adduct detection in aquatic organisms. Among them, functional structure and metabolic traits represent major biological determinants for adducts formed by lipophilic pro-mutagenic contaminants. In detecting DNA adducts through the 32P-postlabelling assay, efficiency in DNA purification, digestion, labelling, as well as adduct enrichment and quantification may explain differences between independent studies. Reference DNA adducts have been used to verify some 32P-postlabelling aspects. Data obtained for mussels and fish treated with benzo[a]pyrene (B[a]P) and environmentally exposed to genotoxins confirm the above assertions. Although the 32P-postlabelling assay cannot be proposed for routine biomonitoring it appears a reliable and very sensitive index of exposure to genotoxic pollutants in both fish and mollusks. 相似文献
7.
DNA single-strand breaks were analysed in the blue mussel (Mytilus edulis L.) deployed in intertidal and subtidal zones in the PAH contaminated Reykjavík harbour and at a reference site, Hvalfj?rethur, Iceland. DNA strand breaks were analysed by Comet assay in isolated gill and haemocyte cells from six mussels from each site and depth. Increased DNA damage in both gill cells and haemocytes were observed in mussels deployed in Reykjavík harbour compared to the reference site. Intertidal mussels from Reykjavík harbour had higher DNA damage in haemocytes compared to subtidal mussels. The Comet assay seems to be useful for measuring genotoxic exposure in mussels from the field, and that DNA damage might be higher in the intertidal zone either due to higher exposure to contaminants or because of physiological and biochemical responses to variations in oxygen availability. 相似文献
8.
Akcha F Tanguy A Leday G Pelluhet L Budzinski H Chiffoleau JF 《Marine environmental research》2004,58(2-5):753-756
DNA single-strand breaks were measured by the comet assay in both gill and hemolymph cells of mussels collected in 3 sampling areas of the French coast (Pointe du Castelli, Pen Bron and Saint-Nazaire Harbour). Whole mussel tissue samples were also collected for the chemical determination of PAH, PCB and heavy metal concentrations. In mussel, a higher level of DNA strand breaks was measured in gill than in hemolymph cells (p < 0.01). Despite a factor of contamination from 2 to 3 between sites, no difference in the extent of mussel DNA strand breaks was shown between sampling locations (p > 0.05), questioning the sensitivity of the assays used in biomonitoring studies. 相似文献
9.
Effects of genotoxic compounds on DNA and development of early and late grass shrimp embryo stages 总被引：1，自引：0，他引：1
Early and late developmental stages of grass shrimp embryos were exposed to different concentrations of two genotoxicants, 2-methyl-1,4-naphthoquinone (MNQ) and 4-nitroquinoline-N-oxide (NQO). DNA strand breaks were assessed by the comet assay while embryo development effects were determined by % of embryos hatching. Early embryo stage embryos were significantly more sensitive to genotoxicants than late stages. For example, all stage 4 embryos failed to hatch at 1 microM NQO while 95% of stage 8 hatched at this concentration. High DNA tail moments, which are a measure of the number of DNA strand breaks, were found in late stage embryos exposed to genotoxicants. Early stage embryo development was effected by low concentrations of genotoxicants but no changes were observed in DNA tail moments. We suggest that high DNA moments in late embryo stages reflect high DNA repair activity, while early stages may lack a fully developed DNA repair system. 相似文献
10.
Lee Shugart 《Marine environmental research》1988,24(1-4)
This paper reports a rapid and facile procedure for obtaining of DNA from small, whole aquatic organisms (fathead minnow, Pimephales promelas) as well as from the liver of another species (bluegill sunfish, Lepomis macrochirus) and the subsequent estimation of its double strandedness. DNA was isolated from various tissues by homogenization in a basic pH solution containing detergent followed by differential extraction using organic solvents. Additional purification was accomplished by molecular sieve chromatography. Under defined alkaline unwinding conditions, the amount of double-stranded and single-stranded DNA was determined using the bisbenzimidazole dye Hoechst 33258. The technique was used to detect the occurrence of strand breaks in DNA of fathead minnows chemically exposed to benzo[a]pyrene. 相似文献
11.
固相萃取-高效液相色谱-荧光检测法测定海水中的多环芳烃 总被引：4，自引：0，他引：4
建立了用固相萃取-高效液相色谱-荧光检测法测定海水中多环芳烃的方法,优化了色谱条件和萃取条件。除苊不能用荧光检测器检出外,其余15种多环芳烃的空白加标回收率在64.5%(苯并[g,h,i],茚并[1,2,3-cd]芘)～88.7%(苯并[a]蒽)之间,相对标准偏差(n=5)为4.9%(荧蒽,苯并[b]荧蒽)～11.1%(苯并[g,h,i],茚并[1,2,3-cd]芘),方法的检出限在0.72(蒽)～14.10 ng/L(荧蒽)之间,基本上达到了痕量分析的要求。利用该方法测得青岛湾表层海水中多环芳烃的浓度在0.125(苯并[k]荧蒽)～25.996 ng/L(萘)之间,但苯并[a]芘未检出。 相似文献
12.
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
13.
The modified nucleoside 7,8-dihydro-8-oxodeoxyguanosine (8-oxo-dG) is an index of oxidative DNA damage. An immunohistochemical approach based on the use of monoclonal antibody 1F7 against 8-oxo-dG was investigated in marine organisms with immunoperoxidase and immunofluorescent detection. Relative staining intensity as a measure of the 8-oxo-dG level was microscopically assessed. After laboratory exposures to benzo[a]pyrene (B[a]P), higher levels of oxidative DNA damage were clearly detected in all treated animals compared to controls. While this method eliminates DNA extraction reducing the processing of biological samples, absolute values are not provided. Further, the method requires only small amounts of tissue and potentially discriminates susceptibility to oxidative damage in different cell types. These results suggest that the assay should have practical applications in marine ecotoxicology. 相似文献
14.
R.H.M. Eertman C.L.F.M.G. Groenink B. Sandee H. Hummel A.C. Smaal 《Marine environmental research》1995,39(1-4)
The polycyclic aromatic hydrocarbons fluoranthene and benzo(a)pyrene significantly reduced the feeding rate of mussels. For both compounds the tissue concentration resulting in a 50% reduction of the clearance rate (TEC50) was calculated. At high tissue concentrations both aromatic compounds reduced the tolerance of mussels to aerial exposure, whereas at low tissue concentrations an improved response was noticed. The activities of the antioxidant enzymes Superoxide dismutase (SOD) and catalase were elevated only at low tissue concentrations of fluoranthene and benzo(a)pyrene. At the highest measured tissue concentrations the activity of both enzymes was reduced, possibly due to a narcotic effect. The reproductive success rate of mussels appeared to be affected negatively by the investigated hydrocarbons. The results of a pilot experiment indicate that mussels can be used also for the testing of contaminated sediments. 相似文献
15.
The assessment of DNA damage by the Comet assay has been described as a useful non-specific general biomarker of stress in many marine organisms. In field situations it has successfully been employed to distinguish between reference and polluted sites and in the laboratory it has been widely used as a mechanistic tool to determine pollutant effects and mechanisms of DNA damage. To date a wide range of marine vertebrates and invertebrates have been used, however, the usefulness of this assay as a biomarker in cnidarians has not yet been assessed. The aims of this study were to optimize the Comet assay for cnidarian cells and to assess its utility for detecting genotoxic damage in these cells. Cells were isolated from the North American pacific coast temperate sea anemone Anthopleura elegantissima using a non-enzymatic dissociation procedure and viability was determined to be in excess of 90%. Cells were incubated either with (1 h acute exposures) hydrogen peroxide (H(2)O(2)), ethylmethanesulphonate (EMS) or benzo(a)pyrene (B[a]P). In comparison to other marine species, anemone cells exhibited high control or background levels of DNA strand breaks. Despite this, however, we observed dose responses for each of the study chemicals with no reduction in cell viability. This study demonstrates that anemone cells respond to known DNA damaging agents, including B[a]P which requires metabolism to exert its genotoxic effect, and that the Comet assay may prove to be a useful biomarker of stress in cnidarian species. 相似文献
16.
夏季珠江口水体中多环芳烃的分布、组成及来源 总被引：2，自引：0，他引：2
利用1999年7月对珠江口海域的调查资料,对该区表层海水中优控多环芳烃的分布、组成及来源进行了分析和讨论,结果表明:(1)夏季珠江口海域表层海水中14种溶解态多环芳烃[苊、芴、菲、蒽、荧蒽、芘、苯并(a)蒽、艹屈、苯并(b)荧蒽、苯并(k)荧蒽、苯并(a)芘、二苯并(a,h)蒽、苯并(g,h,i)艹北、茚并(1,2,3-cd)艹比]的质量浓度为63.8~171.7 ng/L,且沿着冲淡水流向呈降低趋势;(2)颗粒态中15种多环芳烃[萘、苊、芴、菲、蒽、荧蒽、芘、苯并(a)蒽、艹屈、苯并(b)荧蒽、苯并(k)荧蒽、苯并(a)芘、二苯并(a,h)蒽、苯并(g,h,i)艹北、茚并(1,2,3-cd)艹比]的质量浓度为60.7~186.7 ng/L,其分布与水体载沙量及悬浮颗粒物的性质、粒径有关,具有从河口内向外海降低的分布特征;(3)多环芳烃组成和特征参数比值的分析表明,珠江口海域高温裂解来源的多环芳烃在伶仃洋海区输入最多,且主要为人类活动中煤燃烧排放的,而在香港岛周围海区的输入则相对较少,且主要为油燃烧排放的;(4)与法国塞纳河及长江口等河口相比,珠江三角洲海域水体中存在高菲含量排放源。 相似文献
17.
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. 相似文献
18.
19.
The disruption of chromosome segregation which is detected visually by the anaphase aberration (aa) test suggests that unequal amounts of DNA are distributed to daughter cells and possibly to subsequent cell generations. To investigate this possibility trout cell cultures and trout embryos (blastodisc) were exposed to benzo[a]pyrene (B[a]P) and the nitrosamide, MNNG, respectively. They were then examined by flow cytometry to determine if anaphase damage resulted in unequal DNA distribution to daughter cells. Both B[a]P and MNNG produced a significant increase (P < 0·01) in aa in cultured cells after 48 h exposure. These values returned to normal by 10 days in the absence of the genotoxic agents, except for the highest concentration (0·5 μg/ml MNNG), which showed only a 50% recovery by that time. Likewise, the coefficient of variation (CV) of nuclear DNA content of the cells also rose significantly after treatment and remained elevated for as long as 14 days following exposure. Experiments with rainbow trout embryos also showed a significant increase in both aa and CV following exposure to MNNG. Flow cytometric analysis of DNA content of trout cells and embryos following exposure to mutagens showed an unequal distribution of DNA that was transmissible through several cell generations. These findings indicate that visible anaphase aberrations could predict heritable genetic defects such as those associated with aneuploidy. 相似文献
20.
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