Genotoxic assessment of anthracene and benzo [a] pyrene to milkfish Chanos chanos

Mutagenic and genotoxic effects of polycyclic aromatic hydrocarbons, anthracene and benzo [a] pyrene (BaP), in milkfish Chanos chanos were determined using micronucleus (MN) test and comet assay (CA). Distinct mean frequencies of nuclear abnormalities such as MNs; binucleated micronuclei, nuclear bud, and fragmented apoptotic cells were measured. Significant increase in DNA damage with five classes of damage level was observed and expressed in terms of arbitrary unit (AU). Mean frequencies of total nuclear abnormalities were 0.5?±?0.25 cells in control; 0.67?±?0.33 cells in solvent control; 70?±?9.60 cells in 0.176?mg?L^?1 anthracene, and 91.83?±?6.25 cells in 0.031?mg?L^?1 BaP. The greatest DNA damage of 170AU was observed in 0.176?mg?L^?1 anthracene-exposed group and 182AU was observed in 0.031?mg?L^?1 BaP-treated fish. This study confirmed that the CA and MN assays are useful tools in determining potential genotoxicity of water pollutants and might be appropriate as a part of monitoring program.     

Effects of benzo[a]pyrene toxicity on morphology and ultrastructure of Hordeum sativum

Many studies have been devoted to investigation of toxic benzo(a)pyrene (BaP) compound, but studies involving changes at the cellular level are insufficient to understand the mechanisms of polycyclic aromatic hydrocarbons (PAHs) effect on plants. To study the toxicity of BaP, a model vegetation experiment was conducted on cultivation of spring barley (Hordeum sativum distichum) on artificially polluted BaP soil at different concentrations. The article discusses the intake of BaP from the soil into the plant and its effect on the organismic and cellular levels of plant organization. The BaP content in the organs of spring barley was determined by the method of saponification. With an increase in the concentration of BaP in the soil, its content in plants also rises, which leads to inhibition of growth processes. The BaP content in the green part of Hordeum sativum increased from 0.3 µg kg^?1 in control soil up to 2.6 µg kg^?1 and 16.8 µg kg^?1 under 20 and 400 ng/g BaP applying in soil, as well as in roots: 0.9 µg kg^?1, 7.7 µg kg^?1, 42.8 µg kg^?1, respectively. Using light and electron microscopy, changes in the tissues and cells of plants were found and it was established that accumulation of BaP in plant tissues caused varying degrees of ultrastructural damage depending on the concentration of pollutant. BaP had the greatest effect on the root, significant changes were found in it both at histological and cytological levels, while changes in the leaves were observed only at the cytological level. The results provide significant information about the mechanism of action of BaP on agricultural plants.

     

Multiple-time contamination of benzo(a)pyrene in soils inhibits soil enzymatic activities

Benzo[a]pyrene (B[a]P) is accumulating in soils in a low-dose cumulative manner. The objectives of this study were to investigate the effects of B[a]P on the extractable and available fractions of B[a]P and on soil enzymatic activity using multiple-time superimposed and one-time contamination approaches. Results showed that the contents of B[a]P rapidly decreased in the first 14?d and later decreased slowly from 14 to 56?d in both one-time and multiple-time contamination tests. The contents of B[a]P in the multiple-time contamination test were lower than those in the one-time test. Soil urease, sucrase and dehydrogenase activities were rapidly inhibited in the early stage (14?d) and stimulated during the rest of the incubation, and soil dehydrogenase activity was more sensitive to B[a]P contamination than the other enzymes. High concentrations of B[a]P in soil led to greater inhibition of enzymatic activity than that at low concentrations in the early period of culture. Soil enzyme activities were weakly inhibited in multiple-time compared with in one-time contamination tests and were lower in the subsurface layer than in the surface layer. Our results revealed that the multiple-time superimposed approach might be better than one-time contamination for evaluating B[a]P risk in soil.     

Modification of mutagenic activity of benzo[a]pyrene by trace elements in the Ames assay

Benzo[a]pyrene (BaP) is one of the most ubiquitous environmental contaminants and exerts potent carcinogenicity and mutagenicity. In the presence of rat liver S9 mix, four trace elements, germanium (Ge), iron (Fe), zinc (Zn), and molybdenium (Mo), were included in the Ames Salmonella microsome/mutagenic assay to test their inhibitory effects on BaP-induced mutations. All trace elements were found to exert an inhibitory effect in tester TA98 and TA100 strains; Fe was the most potent among the elements tested, whereas Zn was weakly inhibitory to prevent mutations. The present findings demonstrate that the antimutagenic potential of trace elements, notably Fe, may be attributed to its modulatory effect on the bioactivation and detoxification processes of BaP.     

Degradation of Benzo[a]Pyrene in Soil with Arbuscular Mycorrhizal Alfalfa

Mycorrhizal and non-mycorrhizal alfalfa (Medicago sativa) was grown in pots containing soil artificially contaminated with various levels of benzo[a]pyrene (B[a]P)(0, 1, 10 and 100 mg kg^–1). Soil and plants were sampled after 30, 40, 50, 60 and 90 days and compared with unlanted pots. The percentage of mycorrhizal root length colonized by Glomus caledoniun was not significantly affected by the addition of B[a]P up to 10 mg kg^–1 but was significantly lower at 100 mg kg^–1B[a]P compared with low concentrations (p < 0.05). There was no difference in soil polyphenol oxidase and dehydrogenase activity among the controls and applications of 1 and 10 mg kg^–1 of B[a]P. However, enzyme activities were significantly higher at 100 mg kg^–1B[a]P compared with the other three treatments, and there was no mycorrhizal effect. Over a period of 90 days the concentration of B[a]P in soil in which alfalfa was grown was significantly lower than in unplanted soil (p < 0.05). Degradation rates of B[a]P added at 1, 10 and 100 mg kg^–1 without G. caledonium were 76, 78 and 53%, and with mycorrhizal inoculation were 86, 87 and 57%. The degradation rate in unplanted soil was significantly lower than in planted soil, and was significantly higher in medium- and low-B[a]P treatments than in the high B[a]P concentration tested. There is a possibility of enhancement phytoremediation of PAHs in rhizosphere soil with arbuscular mycorrhizal fungi.     

苯并［a］芘对黑鲷肝脏GST活性的影响及其与肝脏代谢酶和胆汁代谢产物之间的变化关系

为探讨苯并[a]芘(B[a]P)暴露对鱼类的影响并筛选特异、敏感的生物标志物,研究了B[a]P对黑鲷(Sparus macrocephalus)肝脏谷胱甘肽-S-转移酶(GST)活性的影响,并分析了肝脏7-乙氧基异吩唑酮-脱乙基酶(EROD)、GST活性与胆汁中B[a]P典型代谢产物3-羟基-苯并[a]芘(3-OHB[a]P)3者之间及与B[a]P之间的剂量、时间-效应关系.结果显示:在B[a]P(0.5、1.0、2.0和5.0μg·L-1)暴露期间,肝脏GST活性随暴露时间总体呈"钟"形的变化趋势,在第2d时达到峰值;在第12h、1d和2d时,GST活性与B[a]P暴露浓度呈显著正相关(R12h=0.966(p<0.05)、R1d=0.953(p<0.05)、R2d=0.824(p<0.05)).与前期研究结果对比分析发现,B[a]P(0.5、1.0、2.0和5.0μg·L-1)暴露7d时,胆汁中3-OHB[a]P浓度与B[a]P暴露浓度呈显著的剂量-效应关系(R=0.943,p<0.05);黑鲷肝脏GST、EROD活性与3-OHB[a]P的对数浓度均呈显著正相关(RGST=0.740(p<0.05)、REROD=0.839(p<0.05));2.0μg·L-1B[a]P暴露后,黑鲷肝脏EROD、GST活性与3-OHB[a]P随暴露时间的变化趋势基本相同,但并不完全一致.鱼类肝脏EROD、GST与胆汁中B[a]P代谢产物3者之间的变化关系较为复杂,暴露浓度和时间是影响其变化的重要因素,肝脏GST和EROD活性比胆汁中代谢产物更易受其影响,在B[a]P的环境监测中代谢产物可能是一种更为可行的生物标志物.     

Benzo [a] pyrene concentrations in topsoils of North Bahrain

A comprehensive field study was conducted to determine the background concentrations of benzo [a] pyrene (B[a]P) in soil samples collected at different points on a grid covering most of the northern and middle parts of the Kingdom of Bahrain. The average B[a]P concentration was 108.4?ng?g^?1 in winter and 73.0?ng?g^?1 in summer. The capital city, Manama, and the industrial areas of the country showed the highest levels of B[a]P.     

苯并(a)芘神经毒性研究进展

苯并(a)芘(BaP)是一种广泛存在于环境中的多环芳烃,具有致癌、致畸、致突变性。目前BaP的神经毒性研究零散而不深入,本文主要总结分析了BaP的神经毒性表现,简要介绍了BaP诱发神经毒性呈现的剂量-效应关系,进一步系统阐述了目前研究中发现的可能分子机制,包括BaP诱导神经递质及其代谢物含量的改变,相关DNA及蛋白质的损伤,抗氧化系统及线粒体的改变等,以期为进一步研究提供参考。     

Distribution,source apportionment and health risk assessment of polycyclic aromatic hydrocarbons (PAHs) in intertidal sediment of Asaluyeh,Persian Gulf

Surface sediment samples were collected from intertidal zone of Asaluyeh, Persian Gulf, to investigate distribution, sources and health risk of sixteen polycyclic aromatic hydrocarbons (PAHs). Total PAH concentrations ranged from 1.8 to 81.2 μg kg^?1 dry weight, which can be categorized as low level of pollution. Qualitative and quantitative assessments showed that PAHs originated from both petrogenic and pyrogenic sources with slight pyrogenic dominance. Source apportionment using principal component analysis indicated that the main sources of PAHs were fossil fuel combustion (33.59%), traffic-related PAHs (32.77%), biomass and coal combustion (18.54%) and petrogenic PAHs (9.31%). According to the results from the sediment quality guidelines, mean effects range-median quotient (M-ERM-Q) and benzo[a]pyrene toxic equivalents (BaP_eq), low negative ecological risks related to PAH compounds would occur in the intertidal zone of Asaluyeh. The total benzo[a]pyrene (BaP) toxic equivalent quotient (TEQ^carc) for carcinogenic compounds ranged from 0.01 to 7 μg kg^?1-BaP_eq, indicating low carcinogenic risk. The human health risk assessment of PAH compounds via ingestion and dermal pathways suggests low and moderate potential risk to human health, respectively.     

固定化微生物对土壤中苯并芘的降解

研究了3株细菌与3株真菌对土壤中苯并芘（BaP）的降解动态,从中筛选出1株细菌（Bacillus sp.）和1株真菌（Mucor sp.）,并采用吸附法将混合菌固定在改性后蛭石上,研究了固定化混合菌对土壤中BaP的降解效果。结果表明：细菌中芽孢杆菌（Bacillus sp.,SB02）降解率最高,42 d对B[a]P的降解率为33.0%,降解速率也最快,1周可降解12.6%的BaP;真菌中毛霉（Mucor sp.,SF06）降解率最高,42 d对B[a]P的降解率为69.7%;以改性后蛭石为载体用吸附法制得的固定化混合菌,传质性能好,对BaP的降解率42 d可达95.32%,高于游离菌20个百分点。     

Laboratory observations on spawning,fecundity and early development of a mesopelagic ostracod,Conchoecia pseudodiscophora,from the Japan Sea

The fate of the polycyclic aromatic hydrocarbon [³H]benzo[a]pyrene (BaP) was examined in two species of scleractinian corals, Favia fragum (Esper) and Montastrea annularis (Ellis and Solander), which were collected in the patch reefs surrounding Alina's Reef (25°23.25N; 80°09.8W) in Biscayne National Park, Florida, USA, in July, 1990. Corals were exposed to initial concentrations of 5 g/l in a simple static system for 25 h. BaP uptake was estimated from the disappearance of BaP from the water. Uptake rates were 6.5±0.7 and 10.8±0.2 g BaP cm^-2h^-1 for F. fragum and M. annularis, respectively, at initial BaP concentrations and were directly proportional to the concentration of BaP in the water. The separation of zooxanthellae from coral tissue revealed that zooxanthellae can accumulate up to 53 and 64% of the total BaP-derived radioactivity present in F. fragum and M. annularis, respectively. Both corals metabolized BaP slowly, as most of the accumulated radioactivity was present as the unmetabolized chemical. However, aqueous and organic-soluble metabolites were found in both the animal and zooxanthellae fractions. Analysis by high-performance liquid chromatography (HPLC) revealed that both species of corals metabolized BaP to various tetrols, triols, dihydrodiols, quinones and phenols, although the pattern of metabolites differed between species. Zooxanthellae contained some of the same Phase I metabolites found in the animal tissue; however, tetrols and triols were absent in extracts from the zooxanthellae. The elimination of BAP from corals was also slow; approximately 38 and 65% of the accumulated radioactivity was still present in F. fragum and M. annularis, respectively, 144 h following the transfer of exposed corals to an uncontaminated flow-through seawater system.     

Benzo[a]pyrene oxidation and microsomal enzyme activity in the mussel (Mytilus edulis) and other bivalve mollusc species from the Western North Atlantic

Analysis of subcellular fractions revealed a complement of microsomal electron transport components including reductases and heme proteins in several organs of the three bivalve species Mytilus edulis, Macrocallista maculata and Area zebra. Dithionite difference spectroscopy of CO-treated microsomes yielded spectra typical of cytochrome P-450 in digestive gland and gill, with absorption maxima at 450 nm. A time-dependent reduction of cytochrome P-450 was also observed. The levels of these components and rates of microsomal benzo[a]pyrene (BP) metabolism were highest in the digestive gland, and were very similar between species. In M. edulis there was a suggested seasonal variation in BP metabolism but no population differences in this activity or in levels of other components. Digestive gland microsomal metabolites of BP identified by HPLC retention and UV spectroscopy included BP-1.6-quinone, BP-3,6-quinone and BP-6,12-quinone, which comprised 65% of the total metabolites, and dihydrodiols and phenols, the latter products consistent with cytochrome P-450 monooxygenation and expoxide hydrolase function. However, the inconsistent dependence of BP metabolism on NADPH, and inconsistent inhibition by CO suggest that catalyst(s) additional to cytochrome P-450 may be acting in BP metabolism. Based on these results and the prominent quinone formation, we speculate that peroxidative mechanism(s) may be involved. The role of peroxidative as well as well as monooxygenase reactions in the in-vivo disposition and effects of foreign chemicals in bivalves, and also the major function of cytochrome P-450 in these bivalves, remain to be established.Some of these results have appeared in preliminary form; J. J. Stegeman, Sea Grant Annual Report, Words Hole Oceanographic Institution, p 15, 1981     

Chemopreventive effects of ellagic acid against genotoxicity induced by benzo(a)pyrene

Benzo(a)pyrene (B(a)P) is a commonly used indicator for polycyclic aromatic hydrocarbons (PAHs) contamination. The use of certain phytochemicals and some polyphenolic compounds proved to be effective in blocking PAH-induced effects. The aim of the present study was to examine the possible inhibitory effects of ellagic acid (EA), present in berries and nuts, against B(a)P-mediated toxicity using in vitro and in vivo test systems. In vitro method included the Ames test, using Salmonella typhimurium tester strain TA100. In vivo experiments were based on mouse bone marrow micronucleus (MN) assay and immunoblotting. EA produced a decrease in the number of revertant colonies against B(a)P in the Ames assay. Further, there was a reduction in the mean number of micronucleated polychromatic erythrocytes in the presence of EA in the co-treatment protocol of the MN test. Western blotting results showed downregulation of CYP450 isoform CYP1A1 which prevented bioactivation of B(a)P. Thus, the present study demonstrates antimutagenic role played by EA against the mutagenic activity of B(a)P in both in vitro and in vivo systems.     

UVR-induced toxicity of sludge and polycyclic aromatic hydrocarbons on seed germination and seedling growth of Triticum aestivum L.

The depletion of the ozone layer and enhancement of solar radiation may exert an adverse influence on the ecosystem. Phototoxicity of sludge and polycyclic aromatic hydrocarbons (PAHs) under ultraviolet radiation (UVR) to seedling was studied. Seeds of wheat (Triticum aestivum) were planted in sludge and PAHs (anthracene (An), benzo(a)pyrene (BaP) and pyrene (Py)) with and without UVR. Toxicity of sludge increased in the presence of UVR in wheat. UVB radiation was found to be more hazardous than UVA radiation. Results demonstrated that An, BaP and Py induced phototoxicity at various concentrations (1–10?µg/mL) under UVA (1.5?mW/cm²) or UVB (1.08?J/cm²) exposure. The pattern of phototoxicity was An?>?Py?>?BaP to shoot length, root length, and fresh weight; chlorophyll, protein content, enzyme activity of catalase and α-amylase were reduced while the activity of superoxide dismutase and starch was enhanced. Reduction in seedling growth and biochemical parameters may be related to less photosynthesis, less nutritional uptake, and distortion of root cap. Thus, the synergistic effect may be due to alterations in photosynthesis, phytohormones, or nutritional uptake.     

Mutagenicity of polyaromatic hydrocarbons by chemical models for cytochrome P450 in Ames assay

DNA damage is an important step in carcinogenesis. The Ames assay is a short-term screening of carcinogens that induce DNA damage. Most carcinogens require enzymatic activation through oxidation by cytochrome P450 (CYP450) in the presence of S9 mix. A combination of iron (Fe)(III) porphyrin and an oxidant is also able to oxidize compounds as an alternative metabolic pathway to CYP450. Previously it was reported that a chemical model containing a water-soluble 5,10,15,20-tetrakis(1-methylpyridinium4-yl)porphyrinatoiron(III) chloride (4-MPy) and tert-butyl hydroperoxide (t-BuOOH) activated aromatic amines and amides. In this study, a chemical model composed of an Fe porphyrin, water-insoluble 5,10,15,20-tetrakis(pentafluorophenyl)porphyrinatoiron(III) chloride (F₅P) or water-soluble 4-MPy was optimized with an oxidant – t-BuOOH, magnesium monoperoxyphthalate (MPPT), or iodosylbenzene (PhIO). Subsequently the mutagenicity of benzo[a]pyrene (B[a]P) and chrysene in Salmonella typhimurium TA strains was compared. B[a]P was activated by a combination of F₅P or 4-MPy plus MPPT or PhIO in S. typhimurium TA1538. The B[a]P-induced mutagenicity with F₅P plus oxidant was higher than 4-MPy plus oxidant. Mutagenicity of chrysene, a tetracyclic aromatic hydrocarbon, was not detected in the presence of F₅P/PhIO in S. typhimurium TA98, but was activated in the presence of F₅P/MPPT. The F₅P/MPPT activated other polyaromatic hydrocarbons (PAH) in the S. typhimurium TA98 assay including dibenz[a,c]anthracene, dibenz[a,h]anthracene, 3-methylcholanthrene, and benzo[a]anthracene. The results indicated that the F₅P/MPPT was the most efficient model for detecting PAH-induced mutagenicity in the Ames assay.     

高分子量多环芳烃--苯并[a]芘的生物降解研究进展

多环芳烃(PAHs)是一类广泛分布于海洋环境中的含有两个以上苯环的有机化学污染物.苯并[a]芘(BaP)是一种由5个苯环组成的分布广泛、致癌性极强的多环芳烃化合物,人们将其作为多环芳烃的指示物,通过研究其在环境中的产生、迁移、转化、降解及毒理作用来判断多环芳烃的污染情况.BaP已成为国内外环境监测的重要指标之一.本文综述了不同环境中BaP的来源与分布,BaP在环境中的行为,微生物对BaP的代谢途径,以及微生物降解BaP的相关限制因子,并结合作者在这方面研究的部分工作与目前国际研究的热点,提出值得进一步探究的有关问题.图4表3参51     

Accumulation and phytoavailability of benzo[a]pyrene in an acid sandy soil

Effects of benzo[a]pyrene (B[a]P) on ryegrass (Lolium perenne L.) growth, plant accumulation and dissipiation of B[a]P in a red sandy soil (Hapli-Udic Argosol) were studied in a pot experiment. The plants were grown for 61 days in soil spiked with B[a]P at 0, 12.5, 25 and 50 mg kg⁻¹. Control pots without plants were also set up. Soil extractable B[a]P, plant shoot and root biomass, and concentrations of B[a]P in plant shoots and roots were determined. Ryegrass biomass was increased by addition of B[a]P and root B[a]P concentrations were significantly correlated with B[a]P application rate, but no such correlation was found for shoot B[a]P concentrations. This indicates that B[a]P enhanced the growth of the ryegrass. The extractable B[a]P concentration in the planted soil was significantly lower than that in the unplanted control soil at the rate of 50 mg B[a]P kg⁻¹. This indicates that ryegrass may help to dissipate B[a]P in soil at concentrations over 50 mg kg⁻¹ soil although the mechanism for this is not understood.     

Gingko biloba extract and cobalt porphyrin additive to remove harmful components from cigarette smoke and reduce its toxicity

Cigarette filters were modified with a combination of gingko biloba extract and cobalt porphyrin (CGC) to remove harmful components from the cigarette smoke and reduce its toxicity. Smoke analysis results indicated that CGC eliminated up to 32% of benzo[a]pyrene (B[a]P), 52% of N-nitrosonornicotine (NNN), 46% of N-nitrosoanabasine (NAB), 35% of 4-(methylnitrosamine)-1-(3-pyridyl)-1-butanone (NNK), 31% of N-nitrosoanatabine (NAT), 30% of gas-phase free radicals, and 33% of solid-phase free radicals. Biological experiments, including the Ames test, neutral red cytotoxicity assay and chronic toxicity, were conducted for both CGC cigarettes and control cigarettes. Results showed that the toxicity of the CGC cigarettes was lower than those of the control cigarettes. The mechanism by which the CGC components could remove harmful components from cigarette smoke is discussed.     

Toxic equivalency factor approach for risk assessment of polycyclic aromatic hydrocarbons

The toxic equivalency factor (TEF) method has been developed to evaluate structurally related compounds, sharing a common mechanism of action. Because certain polycyclic aromatic hydrocarbons (PAHs) appear to fit this requirement and are ubiquitous environmental contaminants, these compounds are TEF candidates. Toxicity is often expressed relative to benzo[a]pyrene (BaP), the reference standard (e.g., TEF = EC50_BaP/EC50_PAH). BaP‐like toxicity or toxic equivalents (TEQs) can then be calculated: TEQ = S(PAH _i × TEF _i ), where PAH _i and TEF _i are the concentration and TEF, respectively, for individual PAH congeners. Representative TEFs for PAHs were compiled from studies measuring carcinogenic potency and surrogate biomarkers. This review examines the application of TEFs to PAHs with established criteria (i.e., demonstrated need, defined chemical grouping, broad toxicological database, endpoint consistency, additivity, common mechanism of action, consensus). Although all criteria are not satisfied (e.g., endpoint inconsistency, nonadditivity) and more rigorous validation studies are needed, the TEF method is useful when limitations are recognized. Refinements of the method might include incorporation of pharmacokinetic factors and segregation of TEFs derived from mammalian vs. nonmammalian systems to increase endopoint consistency. Probabilistic analysis may also prove useful to assess the range of TEF values reported both within and between human health and ecological risk assessments.