Effects of polycyclic aromatic hydrocarbons (PAHs) in environmental pollution on exogenous and oxidative DNA damage (EXPAH project): description of the population under study

The EXPAH project was a molecular epidemiology study whose aims were to evaluate the hypothesis that polycyclic aromatic hydrocarbons (PAHs) are a major source of genotoxic activities of organic mixtures associated with air pollution. Biomarkers of exposure, effects and susceptibility, and oxidative DNA damage were measured in three PAH-exposed populations from Prague (Czech Republic), Kosice (Slovakia) and Sofia (Bulgaria). Control populations were included from each city. In total 356 individuals were enrolled. A questionnaire was used to determine life style/dietary factors. Ambient air exposure was measured by stationary monitoring, and personal exposure monitoring was also carried out. The characteristics of the population are described in this paper together with their personal exposure to carcinogenic PAHs (c-PAHs). The dose of c-PAH exposure was found to vary between the occupationally exposed (e.g. policemen and bus drivers) and the control populations in each country, and also varied from country to country.     

DNA damage in humans exposed to environmental and dietary polycyclic aromatic hydrocarbons

The paper describes recent research on human DNA damage related to environmental and dietary polycyclic aromatic hydrocarbon (PAH) exposures. The study populations either represent general populations of large geographical regions, or their exposure situation may have relevance to the general population. In Silesia, Poland, and Northern Bohemia, Czech Republic, where coal-based industry and domestic heating are the major sources of PAHs, significant differences have been observed in white blood cell DNA adducts and cytogenetic biomarkers between environmentally exposed and rural control populations, and significant seasonal variations of DNA damage have been detected. Bus drivers, traffic policemen and local residents have been involved in biomarker studies in Copenhagen, Athens, Genoa and Cairo, and differences have been measured in the level of DNA damage of urban and rural populations. Burning of smoky coal in unvented homes in Xuan Wei region, China, causes high PAH exposure of residents, which has been reflected in DNA adduct levels in different tissues. Indoor wood burning in open fireplaces did not increase human DNA adduct levels. Oil-well fires left burning in Kuwait after the Persian Gulf war created an unprecedented environmental pollution. However, insignificant environmental PAH levels were measured several miles from these fires. Aromatic and PAH-DNA adduct levels in white blood cells of US Army soldiers were lower during their deployment in Kuwait, than in Fulda, Germany, where they were stationed before and after serving in Kuwait. The contribution of dietary PAH exposure to blood cell DNA adduct levels had been demonstrated in studies in which volunteers consumed heavily charbroiled beef. Environmental tobacco smoke did not cause detectable changes, as measured by 32P-postlabelling, in DNA adduct levels in non-smokers. In the reviewed studies, observed DNA adduct levels were generally in the range of 1 to 10 adducts, and not higher than 40 adducts in 108 nucleotides. Typically, 1.5 to 3-fold differences have been detected in DNA adduct levels between the exposed and control groups.     

Molecular biology approaches for understanding microbial polycyclic aromatic hydrocarbons (PAHs) degradation

The known or suspected hazards of polycyclic aromatic hydrocarbons (PAHs) have provoked enormous concentration and endeavours to relieve or eliminate these precarious compounds from miscellaneous environments including soil, water and air. Among various interventions, biodegradation is an appealing approach for its comparative high efficiency and preferable safety. Microorganisms played crucial role in biodegradation of PAHs. Traditional access mainly including culture-dependent procedures has discovered and isolated PAHs-degrading microorganisms which could be subsequently applied to specific contaminated locus. Although certain progress has been achieved owing to traditional methods, much details in PAHs bioremedation leave pending because of the complexity nature of this process. As the rapid development of biology, molecular techniques such as PCR, fingerprinting technique (mainly DGGE), DNA hybridization technique and gene reporters technique have been intensively applied to gain further insight into the mechanism of PAHs degradation. These techniques not only proved the existence and role of uncultivable microorganisms in the whole population of PAHs degrading related microbials, but also made it possible to revealed the otherwise undetectable complex relationships between multi-microorganism concerned in PAHs biodegradation. Application of such techniques in the field of PAHs biodegradation were reviewed in this article.     

Molecular biology approaches for understanding microbial polycyclic aromatic hydrocarbons (PAHs) degradation

The known or suspected hazards of polycyclic aromatic hydrocarbons (PAHs) have provoked enormous concentration and endeavours to relieve or eliminate these precarious compounds from miscellaneous environments including soil, water and air. Among various interventions, biodegradation is an appealing approach for its comparative high efficiency and preferable safety. Microorganisms played crucial role in biodegradation of PAHs. Traditional access mainly including culture-dependent procedures has discovered and isolated PAHs-degrading microorganisms which could be subsequently applied to specific contaminated locus. Although certain progress has been achieved owing to traditional methods, much details in PAHs bioremedation leave pending because of the complexity nature of this process. As the rapid development of biology, molecular techniques such as PCR, fingerprinting technique (mainly DGGE), DNA hybridization technique and gene reporters technique have been intensively applied to gain further insight into the mechanism of PAHs degradation. These techniques not only proved the existence and role of uncultivable microorganisms in the whole population of PAHs degrading related microbials, but also made it possible to revealed the otherwise undetectable complex relationships between multi-microorganism concerned in PAHs biodegradation. Application of such techniques in the field of PAHs biodegradation were reviewed in this article.     

微生物降解多环芳烃(PAHs)的研究进展

从多环芳烃(PAHs)的降解菌株的筛选、降解机制以及PAHs污染的生物修复等方面介绍了微生物降解PAHs的最新研究进展。     

Biodegradation of selected UV-irradiated and non-irradiated polycyclic aromatic hydrocarbons (PAHs)

Biodegradation of UV-irradiated anthracene, pyrene,benz[a]anthracene,and dibenz[a,h]anthracene was comparedto that of the non-irradiated samples, individuallyand in synthetic mixtures with enrichment cultures.Combined treatment was repeated for individual anthraceneand for the PAH mixture with Sphingomonas sp.strain EPA 505 and Sphingomonas yanoikuyae.Enrichment culture studies were performed on the PAHmixtures in the presence of the main photoproduct ofanthracene, pure 9,10-anthracenedione. Photochemicallypretreated creosote solutions were also subjected tobiodegradation and the results were compared tothose of the non-irradiated solutions. The primaryinterest was on 16 polycyclic aromatic hydrocarbons(PAHs) listed as priority pollutants by European Union(EU) and the United States Environmental ProtectionAgency (USEPA). Irradiation accelerated thebiodegradation onset for anthracene, pyrene, andbenz[a]anthracene when they were treatedindividually. The biodegradation of irradiatedpyrene started with no lag phase andwas complete by 122 h whereas biodegradation of thenon-irradiated sample had a lag of 280 h andresulted in complete degradation by 720 h. Biodegradation ofPAHs was accelerated in synthetic mixtures, especiallyin the presence of pure 9,10-anthracenedione.In general, irradiation had no effect on the biodegradation of PAHsincubated in synthetic mixtures or with pure cultures. Undercurrent experimental conditions, the UV-irradiation invariablyreduced the biodegradation of PAHs in creosote. Based onthe results of the present and previous photochemical-biologicalstudies of PAHs, the influence of the photochemical pretreatmenton the biodegradation is highly dependent on the compoundsbeing treated and other process parameters.     

The unique aromatic catabolic genes in sphingomonads degrading polycyclic aromatic hydrocarbons (PAHs)

Many members of the sphingomonad genus isolated from different geological areas can degrade a wide variety of polycyclic aromatic hydrocarbons (PAHs) and related compounds. These sphingomonads such as Sphingobium yanoikuyae strain B1, Novosphingobium aromaticivorans strain F199, and Sphingobium sp. strain P2 have been found to possess a unique group of genes for aromatic degradation, which are distantly related with those in pseudomonads and other genera reported so far both in sequence homology and gene organization. Genes for aromatics degradation in these sphingomonads are complexly arranged; the genes necessary for one degradation pathway are scattered through several clusters. These aromatic catabolic gene clusters seem to be conserved among many other sphingomonads such as Sphingobium yanoikuyae strain Q1, Sphingomonas paucimobilis strain TNE12, S. paucimobilis strain EPA505, Sphingobium agrestis strain HV3, and Sphingomonas chungbukensis strain DJ77. Furthermore, some genes for naphthalenesulfonate degradation found in Sphingomonas xenophaga strain BN6 also share a high sequence homology with their homologues found in these sphingomonads. On the other hand, protocatechuic catabolic gene clusters found in fluorene-degrading Sphingomonas sp. strain LB126 appear to be more closely related with those previously found in lignin-degrading S. paucimobilis SYK-6 than the genes in this group of sphingomonads. This review summarizes the information on the distribution of these strains and relationships among their aromatic catabolic genes.     

多环芳烃(PAHs)对油菜生长的影响及其积累效应

多环芳烃(PAHs)是持久性有机污染物(POP)之一, 通过大气沉降和污水灌溉能被植物吸收, 对食品生产安全和人类生命健康具有极大威胁。为探究PAHs对蔬菜作物的生长影响及毒害机理, 采用盆栽试验研究了不同浓度的荧蒽(FLU)和苯并[a]芘(B[a]P)单独胁迫下对油菜(Brassica chinensis)生长、生理和品质的影响及在油菜茎叶内的积累。结果表明: FLU和B[a]P在油菜茎叶内的积累量随着土壤中施加浓度的升高而增加, FLU胁迫下各处理间差异显著(p < 0.05), B[a]P胁迫下5.0和10.0 mg·kg^-1时积累量与对照(CK)相比显著(p < 0.05)增加, 10.0 mg·kg^-1时油菜茎叶内的最大B[a]P积累量没有超过我国食品安全标准; FLU和B[a]P对油菜叶长、叶宽和地上生物量的影响都是低浓度促进高浓度抑制; FLU和B[a]P胁迫下与CK相比株高和光合速率(P_n)值都显著(p < 0.05)降低; 对叶绿素含量的影响是低浓度促进高浓度抑制; 从总体来看, FLU胁迫对还原性维生素C (Vc)具有抑制作用, 5.0 mg·kg^-1时Vc含量最低, 而B[a]P胁迫下变化不规律, 在0.5 mg·kg^-1胁迫时与CK相比略有增加, 5.0 mg·kg^-1时含量最低。     

Exposure to environmental polycyclic aromatic hydrocarbons: influences on cellular susceptibility to DNA damage (sampling Kosice and Sofia)

The aim of this study was to investigate a possible influence of occupational exposure to carcinogenic environmental polycyclic aromatic hydrocarbons (c-PAHs) on cellular susceptibility to the induction of the DNA damage. Monitoring was performed and blood samples were collected from two groups of male subjects: occupationally exposed and matched controls. The group exposed to c-PAHs (average age of 35.1 years) consisted of 52 policemen from Košice and 26 policemen and 25 bus drivers (51 altogether) from Sofia. The control group (average age of 36.4 years) consisted of 54 unexposed subjects from Košice and 24 from Sofia. In the investigated groups 52.5% of exposed subjects and 45.3% of control were current smokers. A challenging dose of X-rays (3 Gy) and an alkaline version of the single cell gel electrophoresis (SCGE) assay, known as Comet assay, were used to evaluate levels of induced DNA damage and repair kinetics in isolated human blood lymphocytes. DNA damage detected in lymphocytes prior to or after irradiation did not differ significantly between exposed and unexposed subjects. A significant decrease in repair efficiency due to exposure to PAHs was observed in the exposed individuals from Košice and Sofia, when analysed separately or together. A negative influence of tobacco smoking on the efficiency of DNA repair was observed. Statistically significant differences were found between subgroups stratified according to education level in Sofia: the half times for DNA repair declined with the increasing level of education. These results confirm that environmental exposure to c-PAHs can alter the ability of blood lymphocytes to repair DNA damage and, as a result could potentially lead to effects that are hazardous to human health.     

污灌土壤中多环芳烃（PAHs）的积累与动态变化研究

对污灌土壤中 1 4种多环芳烃的分析表明 ,各灌区土壤中 PAHs的积累一般以渠首最高 ,渠中次之 ,渠尾含量与对照相当 .但在沈抚石油灌区上、中和下游土壤中均有PAHs的积累 .此外 ,水稻生长期污灌可明显增加土壤中 PAHs的总量 ,各单一污染物的增、减趋势有所不同 .     

土壤－植物系统中多环芳烃和重金属的行为研究

对土壤中多环芳烃和重金属的行为研究表明,与对照相比,０—２０ｃｍ以上表土层存在多环芳烃和重金属积累,２０ｃｍ以下土层未发现积累;与春、秋两次采样结果相比,土壤中多环芳烃的含量有所下降,表明土壤微生物对多环芳烃有一定降解作用,且其降解程度与土壤－植物系统的生态结构有关．菲在地下水中检出浓度较高,表明这一污染物有向下迁移的可能性．此外,柳树对土壤中重金属Ｃｄ的积累有明显的削减与净化作用．本研究表明,严格限制污水中多环芳烃和重金属的污染负荷以及设计合理的生态结构是避免多环芳烃和重金属在土壤中积累的关键．     

土壤,植物样品中多环芳烃（PAHs）分析方法研究

土壤、植物和籽实样品分别用四氢呋喃、甲醇、乙酸乙酯以超声技术提取。提取液经旋转浓缩蒸发仪浓缩,经硅胶柱净化后,由高效液相色谱（ＨＰＬＣ）分离,萤光检测分析。对于土壤、植物和籽实样品,其方法回收率根据各个ＰＡＨ化合物的理化性质不同分别为４５．６８－９３．４２、７７．５９－１０８．１３和７９．１１－９８．９６％,结果表明,二氯甲烷、四氢呋喃适合作为土壤样品的提取剂;甲醇、乙酸乙酯分别适合于植物和籽实样     

Effects of environmental air pollution on endogenous oxidative DNA damage in humans

Epidemiological studies conducted in metropolitan areas have demonstrated that exposure to environmental air pollution is associated with increases in mortality. Carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) are the major source of genotoxic activities of organic mixtures associated with respirable particulate matter, which is a constituent of environmental air pollution. In this study,we wanted to evaluate the relationship between exposure to these genotoxic compounds present in the air and endogenous oxidative DNA damage in three different human populations exposed to varying levels of environmental air pollution. As measures of oxidative DNA damage we have determined 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) by liquid chromatography–tandem mass spectrometry (LC–MS/MS) and cyclic pyrimidopurinone N-1,N² malondialdehyde-2′-deoxyguanosine (M₁dG) by the immunoslot blot assay from lymphocyte DNA of participating individuals. The level of endogenous oxidative DNA damage was significantly increased in individuals exposed to environmental air pollution compared to unexposed individuals from Kosice (8-oxodG adducts) and Sofia (M₁dG adducts). However, there was no significant difference in the level of endogenous oxidative DNA and exposure to environmental air pollution in individuals from Prague (8-oxodG and M₁dG adducts) and Kosice (M₁dG adducts). The average level of M₁dG adducts was significantly lower in unexposed and exposed individuals from Kosice compared to those from Prague and Sofia. The average level of 8-oxodG adducts was significantly higher in unexposed and exposed individuals from Kosice compared to those from Prague. A significant increasing trend according to the interaction of c-PAHs exposure and smoking status was observed in levels of 8-oxodG adducts in individuals from Kosice. However, no other relationship was observed for M₁dG and 8-oxodG adduct levels with regard to the smoking status and c-PAH exposure status of the individuals. The conclusion that can be made from this study is that environmental air pollution may alter the endogenous oxidative DNA damage levels in humans but the effect appears to be related to the country where the individuals reside. Genetic polymorphisms of the genes involved in metabolism and detoxification and also differences in the DNA repair capacity and antioxidant status of the individuals could be possible explanations for the variation observed in the level of endogenous oxidative DNA damage for the different populations.     

Multispecies and monoculture rhizoremediation of polycyclic aromatic hydrocarbons (PAHs) from the soil

In this study, we investigated the potential of multispecies rhizoremediation and monoculture rhizoremediation in decontaminating polycyclic aromatic hydrocarbon (PAH) contaminated soil Plant-mediated PAH dissipation was evaluated using monoplanted soil microcosms and soil microcosms vegetated with several different grass species (Brachiaria serrata and Eleusine corocana). The dissipation of naphthalene and fluorene was higher in the "multispecies" vegetated soil compared to the monoplanted and nonplanted control soil. The concentration of naphthalene was undetectable in the multispecies vegetated treatment compared to 96% removal efficiencies in the monoplanted treatments and 63% in the nonplanted control after 10 wk of incubation. Similar removal efficiencies were obtained for fluorene. However, there was no significant difference in the dissipation of pyrene in both the mono- and multispecies vegetated treatments. There also was no significant difference between the dissipation of PAHs in the monoplanted treatments with different grass species. Principle component analysis (PCA) and cluster analysis were used to evaluate functional diversity of the different treatments during phytoremediation of PAHs. Both PCA and cluster analysis revealed differences in the metabolic fingerprints of the PAH contaminated and noncontaminated soils. However, the differences in metabolic diversity between the multispecies vegetated and monoplanted treatments were not clearly revealed. The results suggest that multispecies rhizoremediation using tolerant plant species rather than monoculture rhizoremediation have the potential to enhance pollutant removal in moderately contaminated soils.     

Aerobic bacterial degradation of polycyclic aromatic hydrocarbons (PAHs) and its kinetic aspects

Aerobic bacterial degradation of PAHs is reviewed. Particular attention is paid to its kinetic aspects (rate and specificity). The general concepts of PAH biodegradation in nature and the role of aerobic bacteria in this process are described. The problem of PAH bioavailability and the mechanism of PAH penetration through bacterial cell wall are discussed. The key role of the reaction of PAH hydroxylation in controlling the rate and specificity of PAH biodegradation process is substantiated. The effects of competitive inhibition, intermediate inhibition, cross induction, and cometabolism are considered. The importance of microbial communities for PAH biodegradation in natural ecosystems is shown. The review contains the list of 138 references.     

白洋淀土壤中多环芳烃的分布特征及来源

采用气相色谱质谱联用仪检测了白洋淀表层(0～20 cm)和亚表层(20～30或农田30～40 cm)土壤中16种多环芳烃的含量。结果表明：表层土壤中多环芳烃总量的变化范围为146.0～645.9 ng·g^-1,平均含量为417.4 ng·g^-1;亚表层土壤中多环芳烃总量的变化范围为43.0～394.5 ng·g^-1,平均含量为152.4 ng·g^-1。表层土壤中多环芳烃含量与有机碳含量相关性不显著,亚表层土壤中多环芳烃含量与有机碳含量呈显著正相关(P<0.01),这可能表明了土壤埋藏改造过程中PAHs与土壤有机相结合程度不断加强,以及土壤中不同环数多环芳烃的环境行为差异,总体上看,与高环(≥4环)多环芳烃相比,萘、菲等低环(2～3环)多环芳烃更容易向下层迁移。PAHs的源解析分析表明,白洋淀表层土壤的多环芳烃表现出显著的以生物质和煤燃烧为主的源特征,这与淀区的人为活动,如秸秆燃烧等相关。     

Mutagenic and carcinogenic actions of some polycyclic aromatic hydrocarbons]

The carcinogenic and mutagenic actions of benz(a)pyrene (BP) and its derivatives 6-methyl-, 6-formyl-, 6-chloro-, 6-hydroxy-, 6-acetoxy-, 6-methoxy- and 4(5)-methoxy-BP were studied in mice and bacteria Salmonella typhimurium TA-98 and TA-100, respectively. The potent carcinogenic agents BP, 6-methyl-, 6-formyl-BP proved also mutagenic in bacteria of both strains. Weak carcinogenic compounds (6-chloro-, 6-methoxy-, 4(5)-methoxy-BP) and noncarcinogenic ones (6-acetoxy-, 6-hydroxy-BP) either turned out nonmutagenic or mutagenic in one of two bacterial strains tested. The differences in carcinogenic and mutagenic actions of the substances under study are not relative to the rate of their oxidation in the enzymatic system.     

Binding of polycyclic aromatic hydrocarbons (PAHs) to teleost aryl hydrocarbon receptors (AHRs)

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous, environmental contaminants that pose a potential risk to fish populations. Both field and laboratory studies suggest that exposure of the early life stages of fish to PAH can mimic the embryotoxic effects of the planar halogenated hydrocarbons (PHHs), the most potent of which is 2,3,7,8-tetrachlorodibenzo-p-dioxin. PHH toxicity is mediated by the aryl hydrocarbon receptor (AHR) and PHH potency is predicted by its AHR-binding affinity and CYP1A induction potency. However, the role of the AHR, if any, in mediating the developmental effects of PAH to fish remains unknown. In this study we looked at the AHR binding affinity of a test set of PAH that had been previously ranked for their potency for inducing teleost CYP1A. PAH that induced CYP1A inhibited [3H]TCDD binding to in vitro-expressed AHRs from rainbow trout and the AHR expressed in PLHC-1 fish hepatoma cells. Generally, the relative rank order for AHR binding affinity predicted the rank order of these same PAH for inducing CYP1A reported in other studies. There was a strong, positive relationship between binding to the PLHC-1 AHR (stimulus) and the EC50s for CYP1A induction (response) in whole juvenile trout and in RTL-W1 cells, but EC50s were much higher than expected for a 1:1 stimulus/response relationship. These data show that the ability of PAH to bind to teleost AHR predicts PAH potency for CYP1A induction. If PAH toxicity is receptor-mediated and predicted by induction potencies, we will have a powerful mechanistic-based tool for rapidly assessing the risk of toxicity to fish of PAH from any source.     

Risk assessment of polycyclic aromatic hydrocarbons (PAHs) in sediments from a mixed-use reservoir

Although reservoirs in China are of great significance, very few studies on risk assessment have been reported for reservoirs. This study investigated distribution characteristics, cancer and ecological risks, and source diagnosis of 16 priority polycyclic aromatic hydrocarbons (PAHs) in sediments from Shitou Koumen Reservoir in Jilin Province, China. A total of 12 sediment samples were collected from the reservoir in August (wet season) 2014. Total PAH concentrations in sediment samples ranged from 1294.51 ng/g to 2755.35 ng/g with a mean concentration of 1757.54 ng/g. For individual PAHs, average concentration of Nap was the highest, 800.56 ng/g, while Acy, Fla, BkF, and DahA were undetected in sediment samples. Light PAHs (2–3 rings) accounting for 74.21% was a dominant PAH compositional pattern. Pearson correlation analysis was carried out; results showed that total PAHs was strongly correlated with the highly enriched sedimentary PAHs, and pH was a major factor in controlling PAH distribution. Lifetime cancer risk was employed to assess cancer risk; results indicated that the fish-culturing area was exposed to cancer risk. The molecular diagnostic ratios of isomeric PAHs were applied to identify possible PAH sources; primary PAH sources were identified as oil-related activities, burning agricultural wastes, vehicular emissions, and industrial discharges.