Comparing the removal of polycyclic aromatic hydrocarbons in soil after different bioremediation approaches in relation to the extracellular enzyme activities
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摘要
A 120-day experiment was conducted to compare the removal of polycyclic aromatic hydrocarbons(PAHs) from agricultural soil after natural attenuation(NA), phytoremediation(P), mycoremediation(M), and plant-assisted mycoremediation(PAM) approaches in relation to the extracellular enzyme activities in soil. The NA treatment removed the total soil PAH content negligibly. The P treatment using maize(Zea mays) enhanced only the removal of low and medium molecular PAHs. The Pleurotus ostreatus cultivated on 30–50 mm wood chip substrate used in M treatment was the most successful in the removal of majority PAHs. Therefore,significantly(p < 0.05) highest total PAH removal by 541.4 μg/kg dw(dry weight)(36%) from all tested M treatments was observed. When using the same fungal substrate together with maize in PAM treatment, the total PAH removal was not statistically different from the previous M treatment. However, the maize-assisted mycoremediation treatment significantly boosted fungal biomass, microbial and manganese peroxidase activity in soil which strongly correlated with the removal of total PAHs. The higher PAH removal in that PAM treatment could be reflected in the following post-harvest time. Our suggested M and PAM approaches could be promising in situ bioremediation strategies for PAH-contaminated soils.
A 120-day experiment was conducted to compare the removal of polycyclic aromatic hydrocarbons(PAHs) from agricultural soil after natural attenuation(NA), phytoremediation(P), mycoremediation(M), and plant-assisted mycoremediation(PAM) approaches in relation to the extracellular enzyme activities in soil. The NA treatment removed the total soil PAH content negligibly. The P treatment using maize(Zea mays) enhanced only the removal of low and medium molecular PAHs. The Pleurotus ostreatus cultivated on 30–50 mm wood chip substrate used in M treatment was the most successful in the removal of majority PAHs. Therefore,significantly(p < 0.05) highest total PAH removal by 541.4 μg/kg dw(dry weight)(36%) from all tested M treatments was observed. When using the same fungal substrate together with maize in PAM treatment, the total PAH removal was not statistically different from the previous M treatment. However, the maize-assisted mycoremediation treatment significantly boosted fungal biomass, microbial and manganese peroxidase activity in soil which strongly correlated with the removal of total PAHs. The higher PAH removal in that PAM treatment could be reflected in the following post-harvest time. Our suggested M and PAM approaches could be promising in situ bioremediation strategies for PAH-contaminated soils.
A 120-day experiment was conducted to compare the removal of polycyclic aromatic hydrocarbons(PAHs) from agricultural soil after natural attenuation(NA), phytoremediation(P), mycoremediation(M), and plant-assisted mycoremediation(PAM) approaches in relation to the extracellular enzyme activities in soil. The NA treatment removed the total soil PAH content negligibly. The P treatment using maize(Zea mays) enhanced only the removal of low and medium molecular PAHs. The Pleurotus ostreatus cultivated on 30–50 mm wood chip substrate used in M treatment was the most successful in the removal of majority PAHs. Therefore,significantly(p < 0.05) highest total PAH removal by 541.4 μg/kg dw(dry weight)(36%) from all tested M treatments was observed. When using the same fungal substrate together with maize in PAM treatment, the total PAH removal was not statistically different from the previous M treatment. However, the maize-assisted mycoremediation treatment significantly boosted fungal biomass, microbial and manganese peroxidase activity in soil which strongly correlated with the removal of total PAHs. The higher PAH removal in that PAM treatment could be reflected in the following post-harvest time. Our suggested M and PAM approaches could be promising in situ bioremediation strategies for PAH-contaminated soils.
引文
Andersson,B.E.,Henrysson,T.,1996.Accumulation and degradation of dead-end metabolites during treatment of soil contaminated with polycyclic aromatic hydrocarbons with five strains of white-rot fungi.Appl.Microbiol.Biotechnol.46(5-6),647-652.
Balajil,V.,Arulazhagan,P.,Ebenezer,P.,2014.Enzymatic bioremediation of polyaromatic hydrocarbons by fungal consortia enriched from petroleum contaminated soil and oil seeds.J.Environ.Biol.35(3),521-529.
Baldrian,P.,2009.Microbial enzyme-catalyzed processes in soils and their analysis.Plant Soil Environ.55(9),370-378.
Binet,Ph.,Portal,J.M.,Leyval,C.,2000.Fate of polycyclic aromatic hydrocarbons(PAH)in the rhizosphere and mycorrhizosphere of ryegrass.Plant Soil 227(1-2),207-213.
Bojes,H.K.,Pope,P.G.,2007.Characterization of EPA's 16 priority pollutant polycyclic aromatic hydrocarbons(PAHs)in tank bottom solids and associated contaminated soils at oil exploration and production sites in Texas.Regul.Toxicol.Pharmacol.47(3),288-295.
Bragato,M.,Joshi,K.,Carlson,J.B.,Tenório,J.A.S.,Levendis,Y.A.,2012.Combustion of coal,bagasse and blends thereof:part II:speciation of PAH emissions.Fuel 96,51-58.
?erny,J.,Balík,J.,Kulhánek,M.,?ásová,K.,Nedvěd,V.,2010.Mineral and organic fertilization efficiency in long-term stationary experiments.Plant Soil Environ.56(1),28-36.
Covino,S.,Svobodová,K.,K?esinová,Z.,Petruccioli,M.,Federici,F.,D'Annibale,A.,et al.,2010.In vivo and in vitro polycyclic aromatic hydrocarbons degradation by Lentinus(Panus)tigrinus CBS 577.79.Bioresour.Technol.101(9),3004-3012.
Djajakirana,G.,Joergensen,R.G.,Meyer,B.,1996.Ergosterol and microbial biomass relationship in soil.Biol.Fertil.Soils 22(4),299-304.
Dubrovskaya,E.,Pozdnyakova,N.,Golubev,S.,Muratova,A.,Grinev,V.,Bondarenkova,A.,et al.,2017.Peroxidases from root exudates of Medicago sativa and Sorghum bicolor:catalytic properties and involvement in PAH degradation.Chemosphere169,224-232.
Dupuy,J.,Ouvrard,S.,Leglize,P.,Sterckeman,T.,2015.Morphological and physiological responses of maize(Zea mays)exposed to sand contaminated by phenanthrene.Chemosphere 124,110-115.
Dvo?ák,T.,Száková,J.,Vondrá?ková,S.,Ko?ná?,Z.,Hole?ková,Z.,Najmanová,J.,et al.,2017.Content of inorganic and organic pollutants and their mobility in bottom sediment from the Orlík water reservoir(Vltava river,Czech Republic).Soil Sediment Contam.26(6),584-604.
Eggen,T.,Majcherczyk,A.,1998.Removal of polycyclic aromatic hydrocarbons(PAH)in contaminated soil by white rot fungus Pleurotus ostreatus.Int.Biodeterior.Biodegrad.41(2),111-117.
Feng,L.,Zhang,L.,Feng,L.,2014.Dissipation of polycyclic aromatic hydrocarbons in soil amended with sewage sludge compost.Int.Biodeterior.Biodegrad.95,200-207.
Feng,N.-X.,Yua,J.,Zhao,H.-M.,Cheng,Y.-T.,Mo,C.-H.,Cai,Q.-Y.,et al.,2017.Efficient phytoremediation of organic contaminants in soils using plant-endophyte partnerships.Sci.Total Environ.583,352-368.
Fu,D.,Teng,Y.,Shen,Y.,Sun,M.,Tu,C.,Luo,Y.,et al.,2012.Dissipation of polycyclic aromatic hydrocarbons and microbial activity in a field soil planted with perennial ryegrass.Front.Environ.Sci.Eng.6(3),330-335.
García-Delgado,C.,Alfaro-Barta,I.,Eymar,E.,2015.Combination of biochar amendment and mycoremediation for polycyclic aromatic hydrocarbons immobilization and biodegradation in creosote-contaminated soil.J.Hazard.Mater.285,259-266.
García-Sánchez,M.,Ko?ná?,Z.,Mercl,F.,Aranda,E.,Tlusto?,P.,2018.A comparative study to evaluate natural attenuation,mycoaugmentation,phytoremediation,and microbial-assisted phytoremediation strategies for the bioremediation of an aged PAH-polluted soil.Ecotoxicol.Environ.Saf.147,165-174.
Gerhardt,K.E.,Gerwing,P.D.,Greenberg,B.M.,2017.Opinion:taking phytoremediation from proven technology to accepted practice.Plant Sci.256,170-185.
Kadri,T.,Rouissi,T.,Brar,S.K.,Cledon,M.,Sarma,S.,Verma,M.,2017.Biodegradation of polycyclic aromatic hydrocarbons(PAHs)by fungal enzymes:a review.J.Environ.Sci.51,52-74.
Ko?ná?,Z.,Tlusto?,P.,2018.Removal of soil polycyclic aromatic hydrocarbons derived from biomass fly ash by plants and organic amendments.Plant Soil Environ.64(2),88-94.
Ko?ná?,Z.,Mercl,F.,Perná,I.,Tlusto?,P.,2016.Investigation of polycyclic aromatic hydrocarbon content in fly ash and bottom ash of biomass incineration plants in relation to the operating temperature and unburned carbon content.Sci.Total Environ.563-564,53-61.
Ko?ná?,Z.,Mercl,F.,Tlusto?,P.,2018.Ability of natural attenuation and phytoremediation using maize(Zea mays L.)to decrease soil contents of polycyclic aromatic hydrocarbons(PAHs)derived from biomass fly ash in comparison with PAHs-spiked soil.Ecotoxicol.Environ.Saf.153,16-22.
Kuppusamy,S.,Thavamani,P.,Venkateswarlu,K.,Lee,Y.B.,Naidu,R.,Megharaj,M.,2017.Remediation approaches for polycyclic aromatic hydrocarbons(PAHs)contaminated soils:technological constraints,emerging trends and future directions.Chemosphere 168,944-968.
Larsson,M.,Hagberg,J.,Rotander,A.,van Bavel,B.,Engwall,M.,2013.Chemical and bioanalytical characterisation of PAHs in risk assessment of remediated PAH-contaminated soils.Environ.Sci.Pollut.Res.20(12),8511-8520.
Li,X.Z.,Wu,Y.C.,Lin,X.G.,Zhang,J.,Zeng,J.,2012.Dissipation of polycyclic aromatic hydrocarbons(PAHs)in soil microcosms amended with mushroom cultivation substrate.Soil Biol.Biochem.47,191-197.
Li,Q.,Li,Y.,Zhu,L.,Xing,B.,Chen,B.,2017.Dependence of plant uptake and diffusion of polycyclic aromatic hydrocarbons on the leaf surface morphology and micro-structures of cuticular waxes.Sci.Rep.7,46235.
Lin,Q.,Shen,K.-L.,Zhao,H.-M.,Li,W.-H.,2008.Growth response of Zea mays L.in pyrene-copper co-contaminated soil and the fate of pollutants.J.Hazard.Mater.150(3),515-521.
Maliszewska-Kordybach,B.,Smreczak,B.,Klimkowicz-Pawlas,A.,2009.Concentrations,sources,and spatial distribution of individual polycyclic aromatic hydrocarbons(PAHs)in agricultural soils in the Eastern part of the EU:Poland as a case study.Sci.Total Environ.407,3746-3753.
Marco-Urrea,E.,García-Romera,I.,Aranda,E.,2015.Potential of non-ligninolytic fungi in bioremediation of chlorinated and polycyclic aromatic hydrocarbons.New Biotechnol.32(6),620-628.
Mercl,F.,Tejnecky,V.,Száková,J.,Tlusto?,P.,2016.Nutrient dynamics in soil solution and wheat response after biomass ash amendments.Agron.J.108(6),2222-2234.
Oleszczuk,P.,Baran,S.,2003.Optimization of ultrasonic extraction of polycyclic aromatic hydrocarbons from sewage sludge samples.Chem.Anal.48(2),211-221.
Public Notice No.153/2016 about the Agricultural Soil Quality and Protection Requirements.Legal code of The Czech Republic,pp.2692-2699.
Public notice No.53/2012 about the chemical requirements of foodstuff and other materials on health protection.Legal code of the Czech Republic,pp.866-984.
Sack,U.,Heinze,T.M.,Deck,J.,Cerniglia,C.R.,Martens,R.,Zadrazil,F.,et al.,1997.Comparison of phenanthrene and pyrene degradation by different wood-decaying fungi.Appl.Environ.Microbiol.63(10),3919-3925.
Shi,W.,Zhang,X.-N.,Jia,H.-B.,Feng,S.-D.,Yang,Z.-X.,Zhao,O.-Y.,et al.,2017.Effective remediation of aged HMW-PAHs polluted agricultural soil by the combination of Fusarium sp.and smooth bromegrass(Bromus inermis Leyss.).J.Integr.Agric.16,199-209.
Smith,M.J.,Flowers,T.H.,Duncan,H.J.,Alder,J.,2006.Effect of polycyclic aromatic hydrocarbons on germination and subsequent growth of grasses and legumes in freshly contaminated soil and soil with aged PAHs residues.Environ.Pollut.141(3),519-525.
Song,Y.F.,Jing,X.,Fleischmann,S.,Wilcke,B.-M.,2002.Comparative study of extraction methods for the determination of PAHs from contaminated soils and sediments.Chemosphere 48(9),993-1001.
?tursová,M.,Baldrian,P.,2011.Effects of soil properties and management on the activity of soil organic matter transforming enzymes and the quantification of soil-bound and free activity.Plant Soil 338(1-2),99-110.
US EPA,1996.Method 3630C:Silica Gel Cleanup,part of Test Methods for Evaluating Solid waste,Physical/Chemical Methods.EPA Publication SW-846,Washinghton,DCDecember,1996.Revision 3-Final Update.Washinghton,DC.February,2007.Revision 3-Final Update.Available:.https://www.epa.gov/hw-sw846/sw-846-test-method-3630c-silica-gel-cleanup,Accessed date:7 January 2018.
US EPA,2007.Method 3550C:Ultrasonic Extraction,Part of Test Methods for Evaluating Solid Waste,Physical/Chemical Methods.EPA Publication SW-846,Washinghton,DC February,2007.Revision 3-Final Update.Available:.https://www.epa.gov/sites/production/files/2015-12/documents/3550c.pdf,Accessed date:7 January 2018.
US EPA,2014.Method 8270D:Semivolatile Organic Compounds by Gas Chromatography/Mass Spectroscopy,Part of Test Methods for Evaluating Solid Waste,Physical/Chemical Methods.EPA Publication SW-846,Washington,DC Revision 5,July,2014-Final Update V.Available:.https://www.epa.gov/hw-sw846/sw-846-test-method-8270d-semivolatile-organiccompounds-gas-chromatographymass-spectrometry,Accessed date:7 January 2018.
Vácha,R.,Horváthová,V.,Vyslou?ilová,M.,2005.The application of sludge on agriculturally used soils and the problem of persistent organic pollutants.Plant Soil Environ.51(1),12-18.
Wang,C.,Sun,H.,Li,J.,Li,Y.,Zhang,Q.,2009.Enzyme activities during degradation of polycyclic aromatic hydrocarbons by white rot fungus Phanerochaete chrysosporium in soils.Chemosphere 77(6),733-738.
Wang,H.,Li,Y.,Xia,X.,Xiong,X.,2018.Relationship between metabolic enzyme activities and bioaccumulation kinetics of PAHs in zebrafish(Danio rerio).J.Environ.Sci.65,43-52.
Wei,Y.-L.,Bao,L.-J.,Wu,C.-C.,He,Z.-C.,Zeng,E.Y.,2014.Association of soil polycyclic aromatic hydrocarbon levels and anthropogenic impacts in a rapidly urbanizing region:spatial distribution,soil-air exchange and ecological risk.Sci.Total Environ.473-474,676-684.
Balajil,V.,Arulazhagan,P.,Ebenezer,P.,2014.Enzymatic bioremediation of polyaromatic hydrocarbons by fungal consortia enriched from petroleum contaminated soil and oil seeds.J.Environ.Biol.35(3),521-529.
Baldrian,P.,2009.Microbial enzyme-catalyzed processes in soils and their analysis.Plant Soil Environ.55(9),370-378.
Binet,Ph.,Portal,J.M.,Leyval,C.,2000.Fate of polycyclic aromatic hydrocarbons(PAH)in the rhizosphere and mycorrhizosphere of ryegrass.Plant Soil 227(1-2),207-213.
Bojes,H.K.,Pope,P.G.,2007.Characterization of EPA's 16 priority pollutant polycyclic aromatic hydrocarbons(PAHs)in tank bottom solids and associated contaminated soils at oil exploration and production sites in Texas.Regul.Toxicol.Pharmacol.47(3),288-295.
Bragato,M.,Joshi,K.,Carlson,J.B.,Tenório,J.A.S.,Levendis,Y.A.,2012.Combustion of coal,bagasse and blends thereof:part II:speciation of PAH emissions.Fuel 96,51-58.
?erny,J.,Balík,J.,Kulhánek,M.,?ásová,K.,Nedvěd,V.,2010.Mineral and organic fertilization efficiency in long-term stationary experiments.Plant Soil Environ.56(1),28-36.
Covino,S.,Svobodová,K.,K?esinová,Z.,Petruccioli,M.,Federici,F.,D'Annibale,A.,et al.,2010.In vivo and in vitro polycyclic aromatic hydrocarbons degradation by Lentinus(Panus)tigrinus CBS 577.79.Bioresour.Technol.101(9),3004-3012.
Djajakirana,G.,Joergensen,R.G.,Meyer,B.,1996.Ergosterol and microbial biomass relationship in soil.Biol.Fertil.Soils 22(4),299-304.
Dubrovskaya,E.,Pozdnyakova,N.,Golubev,S.,Muratova,A.,Grinev,V.,Bondarenkova,A.,et al.,2017.Peroxidases from root exudates of Medicago sativa and Sorghum bicolor:catalytic properties and involvement in PAH degradation.Chemosphere169,224-232.
Dupuy,J.,Ouvrard,S.,Leglize,P.,Sterckeman,T.,2015.Morphological and physiological responses of maize(Zea mays)exposed to sand contaminated by phenanthrene.Chemosphere 124,110-115.
Dvo?ák,T.,Száková,J.,Vondrá?ková,S.,Ko?ná?,Z.,Hole?ková,Z.,Najmanová,J.,et al.,2017.Content of inorganic and organic pollutants and their mobility in bottom sediment from the Orlík water reservoir(Vltava river,Czech Republic).Soil Sediment Contam.26(6),584-604.
Eggen,T.,Majcherczyk,A.,1998.Removal of polycyclic aromatic hydrocarbons(PAH)in contaminated soil by white rot fungus Pleurotus ostreatus.Int.Biodeterior.Biodegrad.41(2),111-117.
Feng,L.,Zhang,L.,Feng,L.,2014.Dissipation of polycyclic aromatic hydrocarbons in soil amended with sewage sludge compost.Int.Biodeterior.Biodegrad.95,200-207.
Feng,N.-X.,Yua,J.,Zhao,H.-M.,Cheng,Y.-T.,Mo,C.-H.,Cai,Q.-Y.,et al.,2017.Efficient phytoremediation of organic contaminants in soils using plant-endophyte partnerships.Sci.Total Environ.583,352-368.
Fu,D.,Teng,Y.,Shen,Y.,Sun,M.,Tu,C.,Luo,Y.,et al.,2012.Dissipation of polycyclic aromatic hydrocarbons and microbial activity in a field soil planted with perennial ryegrass.Front.Environ.Sci.Eng.6(3),330-335.
García-Delgado,C.,Alfaro-Barta,I.,Eymar,E.,2015.Combination of biochar amendment and mycoremediation for polycyclic aromatic hydrocarbons immobilization and biodegradation in creosote-contaminated soil.J.Hazard.Mater.285,259-266.
García-Sánchez,M.,Ko?ná?,Z.,Mercl,F.,Aranda,E.,Tlusto?,P.,2018.A comparative study to evaluate natural attenuation,mycoaugmentation,phytoremediation,and microbial-assisted phytoremediation strategies for the bioremediation of an aged PAH-polluted soil.Ecotoxicol.Environ.Saf.147,165-174.
Gerhardt,K.E.,Gerwing,P.D.,Greenberg,B.M.,2017.Opinion:taking phytoremediation from proven technology to accepted practice.Plant Sci.256,170-185.
Kadri,T.,Rouissi,T.,Brar,S.K.,Cledon,M.,Sarma,S.,Verma,M.,2017.Biodegradation of polycyclic aromatic hydrocarbons(PAHs)by fungal enzymes:a review.J.Environ.Sci.51,52-74.
Ko?ná?,Z.,Tlusto?,P.,2018.Removal of soil polycyclic aromatic hydrocarbons derived from biomass fly ash by plants and organic amendments.Plant Soil Environ.64(2),88-94.
Ko?ná?,Z.,Mercl,F.,Perná,I.,Tlusto?,P.,2016.Investigation of polycyclic aromatic hydrocarbon content in fly ash and bottom ash of biomass incineration plants in relation to the operating temperature and unburned carbon content.Sci.Total Environ.563-564,53-61.
Ko?ná?,Z.,Mercl,F.,Tlusto?,P.,2018.Ability of natural attenuation and phytoremediation using maize(Zea mays L.)to decrease soil contents of polycyclic aromatic hydrocarbons(PAHs)derived from biomass fly ash in comparison with PAHs-spiked soil.Ecotoxicol.Environ.Saf.153,16-22.
Kuppusamy,S.,Thavamani,P.,Venkateswarlu,K.,Lee,Y.B.,Naidu,R.,Megharaj,M.,2017.Remediation approaches for polycyclic aromatic hydrocarbons(PAHs)contaminated soils:technological constraints,emerging trends and future directions.Chemosphere 168,944-968.
Larsson,M.,Hagberg,J.,Rotander,A.,van Bavel,B.,Engwall,M.,2013.Chemical and bioanalytical characterisation of PAHs in risk assessment of remediated PAH-contaminated soils.Environ.Sci.Pollut.Res.20(12),8511-8520.
Li,X.Z.,Wu,Y.C.,Lin,X.G.,Zhang,J.,Zeng,J.,2012.Dissipation of polycyclic aromatic hydrocarbons(PAHs)in soil microcosms amended with mushroom cultivation substrate.Soil Biol.Biochem.47,191-197.
Li,Q.,Li,Y.,Zhu,L.,Xing,B.,Chen,B.,2017.Dependence of plant uptake and diffusion of polycyclic aromatic hydrocarbons on the leaf surface morphology and micro-structures of cuticular waxes.Sci.Rep.7,46235.
Lin,Q.,Shen,K.-L.,Zhao,H.-M.,Li,W.-H.,2008.Growth response of Zea mays L.in pyrene-copper co-contaminated soil and the fate of pollutants.J.Hazard.Mater.150(3),515-521.
Maliszewska-Kordybach,B.,Smreczak,B.,Klimkowicz-Pawlas,A.,2009.Concentrations,sources,and spatial distribution of individual polycyclic aromatic hydrocarbons(PAHs)in agricultural soils in the Eastern part of the EU:Poland as a case study.Sci.Total Environ.407,3746-3753.
Marco-Urrea,E.,García-Romera,I.,Aranda,E.,2015.Potential of non-ligninolytic fungi in bioremediation of chlorinated and polycyclic aromatic hydrocarbons.New Biotechnol.32(6),620-628.
Mercl,F.,Tejnecky,V.,Száková,J.,Tlusto?,P.,2016.Nutrient dynamics in soil solution and wheat response after biomass ash amendments.Agron.J.108(6),2222-2234.
Oleszczuk,P.,Baran,S.,2003.Optimization of ultrasonic extraction of polycyclic aromatic hydrocarbons from sewage sludge samples.Chem.Anal.48(2),211-221.
Public Notice No.153/2016 about the Agricultural Soil Quality and Protection Requirements.Legal code of The Czech Republic,pp.2692-2699.
Public notice No.53/2012 about the chemical requirements of foodstuff and other materials on health protection.Legal code of the Czech Republic,pp.866-984.
Sack,U.,Heinze,T.M.,Deck,J.,Cerniglia,C.R.,Martens,R.,Zadrazil,F.,et al.,1997.Comparison of phenanthrene and pyrene degradation by different wood-decaying fungi.Appl.Environ.Microbiol.63(10),3919-3925.
Shi,W.,Zhang,X.-N.,Jia,H.-B.,Feng,S.-D.,Yang,Z.-X.,Zhao,O.-Y.,et al.,2017.Effective remediation of aged HMW-PAHs polluted agricultural soil by the combination of Fusarium sp.and smooth bromegrass(Bromus inermis Leyss.).J.Integr.Agric.16,199-209.
Smith,M.J.,Flowers,T.H.,Duncan,H.J.,Alder,J.,2006.Effect of polycyclic aromatic hydrocarbons on germination and subsequent growth of grasses and legumes in freshly contaminated soil and soil with aged PAHs residues.Environ.Pollut.141(3),519-525.
Song,Y.F.,Jing,X.,Fleischmann,S.,Wilcke,B.-M.,2002.Comparative study of extraction methods for the determination of PAHs from contaminated soils and sediments.Chemosphere 48(9),993-1001.
?tursová,M.,Baldrian,P.,2011.Effects of soil properties and management on the activity of soil organic matter transforming enzymes and the quantification of soil-bound and free activity.Plant Soil 338(1-2),99-110.
US EPA,1996.Method 3630C:Silica Gel Cleanup,part of Test Methods for Evaluating Solid waste,Physical/Chemical Methods.EPA Publication SW-846,Washinghton,DCDecember,1996.Revision 3-Final Update.Washinghton,DC.February,2007.Revision 3-Final Update.Available:.https://www.epa.gov/hw-sw846/sw-846-test-method-3630c-silica-gel-cleanup,Accessed date:7 January 2018.
US EPA,2007.Method 3550C:Ultrasonic Extraction,Part of Test Methods for Evaluating Solid Waste,Physical/Chemical Methods.EPA Publication SW-846,Washinghton,DC February,2007.Revision 3-Final Update.Available:.https://www.epa.gov/sites/production/files/2015-12/documents/3550c.pdf,Accessed date:7 January 2018.
US EPA,2014.Method 8270D:Semivolatile Organic Compounds by Gas Chromatography/Mass Spectroscopy,Part of Test Methods for Evaluating Solid Waste,Physical/Chemical Methods.EPA Publication SW-846,Washington,DC Revision 5,July,2014-Final Update V.Available:.https://www.epa.gov/hw-sw846/sw-846-test-method-8270d-semivolatile-organiccompounds-gas-chromatographymass-spectrometry,Accessed date:7 January 2018.
Vácha,R.,Horváthová,V.,Vyslou?ilová,M.,2005.The application of sludge on agriculturally used soils and the problem of persistent organic pollutants.Plant Soil Environ.51(1),12-18.
Wang,C.,Sun,H.,Li,J.,Li,Y.,Zhang,Q.,2009.Enzyme activities during degradation of polycyclic aromatic hydrocarbons by white rot fungus Phanerochaete chrysosporium in soils.Chemosphere 77(6),733-738.
Wang,H.,Li,Y.,Xia,X.,Xiong,X.,2018.Relationship between metabolic enzyme activities and bioaccumulation kinetics of PAHs in zebrafish(Danio rerio).J.Environ.Sci.65,43-52.
Wei,Y.-L.,Bao,L.-J.,Wu,C.-C.,He,Z.-C.,Zeng,E.Y.,2014.Association of soil polycyclic aromatic hydrocarbon levels and anthropogenic impacts in a rapidly urbanizing region:spatial distribution,soil-air exchange and ecological risk.Sci.Total Environ.473-474,676-684.