بررسی تاثیر سینرژیستیک گیاه‌پالایی و ایزوله باکتریایی در حذف هیدروکربن‌های آروماتیک حلقوی (PAHs) از خاک آلوده
محورهای موضوعی : ژنتیکمهدی خزائی 1 , علیرضا اطمینان 2 , سولماز دشتی 3 , سیداحمد حسینی 4
1 - گروه محیط زیست و منابع طبیعی، واحد کرمانشاه، دانشگاه آزاد اسلامی، کرمانشاه، ایران.
2 - گروه اصلاح نباتات و بیوتکنولوژی، واحد کرمانشاه، دانشگاه آزاد اسلامی، کرمانشاه، ایران.
3 - گروه محیط زیست، واحد اهواز، دانشگاه آزاد اسلامی، اهواز، ایران.
4 - گروه محیط زیست و منابع طبیعی، واحد کرمانشاه، دانشگاه آزاد اسلامی، کرمانشاه، ایران.
کلید واژه: باکتری, گیاه‌پالایی, سینرژیستیک, هیدروکربن‌های آروماتیک حلقوی,
چکیده مقاله :
هیدروکربن های آروماتیک سرطان زا و سمی بوده و آسیب های جدی به محیط زیست و موجودات زنده وارد می کنند. این ترکیبات عموماً به خاک ها تخلیه می شوند. روش های بیولوژیکی با بکارگیری میکروارگانیسم های موثر جدا شده از خاک های آلوده نفتی و گیاهان مقاوم برای پاکسازی این خاک ها ترجیح داده می شوند. هدف مطالعه حاضر بررسی اثر گیاه کنوکارپوس (Conocarpus erectus) و باکتری سودوموناس آئروژینوزا (Pseudomonas aeruginosa) بر میزان حذف Polycyclic Aromatic hydrocarbons (PAHs) از خاک های آلوده و شور حوضچه تبخیر واحد نمکزدایی در طول 275 روز و در شرایط غیرآزمایشگاهی بود. آزمایش بصورت فاکتوریل با دو فاکتور، در قالب طرح پایه کاملاً تصادفی با سه تکرار طراحی شد. فاکتورهای آزمایش شامل4 سطح (گیاه، باکتری، گیاه و باکتری و خاک بدون کاربرد گیاه و باکتری به عنوان شاهد) و غلظت آلاینده در 5 سطح نفت خام بنگستان با غلظت های صفر، 5/0، 1، 5/2 و 5 درصد وزنی بود. با افزایش غلظت هیدروکربن در هر 5 سطح، درصد حذف و نیز وزن خشک ریشه، اندام هوایی گیاه و کلروفیل کاهش یافت. بالاترین درصد حذف در دو غلظت صفر و1 درصد با درصدهای 43/99، 89/59 و 01/57 درصد به تیمارهای دارای باکتری و گیاه، تیمارهای جداگانه باکتری و گیاه تعلق داشت.. کارایی گیاه و باکتری در حذف هیدروکربن های نفتی تقریباً برابر بود. تیمارهای دارای باکتری در مقایسه با تیمارهای فاقد آن کلروفیل، وزن خشک ساقه و اندام هوایی بیشتری داشتند .نتایج نشان داد که حضور گیاه و باکتری به صورت جداگانه اثر مثبتی برتجزیه میزان PAHs داشت اما اثر سینرژیستیک باکتری و گیاه در تجزیه میزان PAHs مثبت تر بود (05/0P<).
Having carcinogenic and toxic effects, aromatic hydrocarbons cause serious damage to the environment and living organisms. These compounds are mainly discharged into the soil. For the remediation of contaminated soils, biological methods utilizing the efficient microorganisms isolated from the oil-contaminated soils as well as resistant plants are preferred. The aim of this study was to assess the effect of Conocarpus erectus and Pseudomonas aeruginosa on the removal efficiency of polycyclic aromatic hydrocarbons (PAHs) from contaminated and saline soils of the salt separation pond of a desalination unit during 275 days under non-laboratory condition. The study was conducted in a factorial experiment with two factors based on completely randomized design with three replications. The factors used in this experiment included four treatment types (plant, bacteria, plant-bacteria cultivated in the soil, and soil with no plant and bacteria (control)) and the concentration of contaminant (Bangestan crude oil) with 5 levels (0, 0.5, 1, 2.5, and 5 wt%). As hydrocarbon concentrations increased at all five levels, the percentage of PAHs removal, the dry weight of roots and shoots, and chlorophyll contents decreased. At 0 and 1 % concentrations, the highest percentages of removal were obtained as 99.43, 59.89, and 57.01 for bacteria-plant treatment and separate bacterial and plant treatments, respectively (p≤0.05). The plant and the bacteria showed almost equal efficiency in the removal of oil hydrocarbons (p≤0.05). Bacterial treatments led to increased chlorophyll content as well as higher dry weight of roots and shoots compared with the treatments without bacteria (p≤0.05). Results indicated that individual treatments of plant and bacteria had a positive effect on the decomposition rate of PAHs. However, the rate was more positively influenced by the synergistic activity of the bacteria and plants (p≤0.05).
Abena, M., T., Li, Shah, M. and Zhong, W. (2019). Biodegradation of total petroleum hydrocarbons (TPH) in highly contaminated soils by natural attenuation and bioaugmentation. Chemosphere. 234: 864-874.
Al-Hawas, G.H.S., W.M. Sukry, Azzoz, M.M. and Al-Moaik. R.M.S. (2012). The effect of sublethal concentrations of crude oil on the metabolism of Jojoba (Simmodsia chinensis) seedlings. International Research Journal of Plant Science. 3(4):54-62.
Bandowe, B. A. M., S. Leimer, Meusel, H. and Velescu, A. (2019). Plant diversity enhances the natural attenuation of polycyclic aromatic compounds (PAHs and oxygenated PAHs) in grassland soils. Soil Biology and Biochemistry. 129: 60-70.
Baruah, P., Saikia, R.R., Baruah, P.P. and Deka, S. (2014). Effect of crude oil contamination on the chlorophyll content and morpho-anatomy of Cyperus brevifolius (Rottb.). Environmental Science and Pollution Research. 21:12530–12538.
Bisht, S., Pandey, P., Bhargava, B.S., Sharma, Kumar, V. and Sharma, K.D. (2015). Bioremediation of polyaromatic hydrocarbons (PAHs) using rhizosphere technology. Brazilian Journal of Microbiology. 46: 7-21
Chiou, C., Sheng, G. and Manes, M. (2001). A partition-limited model for the plant uptake of organic contaminants from soil and water. Environmental Science and Technology. 35(7): 1437–44.
Chukwuma, C., Ikewuchi, J. and Monanu, M. (2019). Removal of hydrocarbons from crude oil contaminated agricultural soil by phytoremediation using Mariscus alternifolius and Fimbristylis ferruginea. European Journal of Biological Research. 9(1): 34-44.
D’Orazio, V., Ghanem, A. and Senesi, N. (2013). Phytoremediation of pyrene contaminated soils by different plant species. CLEAN–Soil. Air., Water. 41(4):377-82
Dasgupta, D., Jublee, J. and Suparna, M. (2018). Characterization, phylogenetic distribution and evolutionary trajectories of diverse hydrocarbon degrading microorganisms isolated from refinery sludge. Biotechnology. 8: 273-282.
Doyle, E., L. Muckian, Hickey, A.M. and Clipson, N. (2008). Microbial PAH degradation. Advances in Applied Microbiology. 65: 27–66
Dudhagara D.R., R.K. Rajpara, J.K., Bhatt, H.B., Gosai, Sachaniya, B.K. and Dave, B.P. (2016). Distribution, sources and ecological risk assessment of PAHs in historically contaminated surface sediments at Bhavnagar coast, Gujarat, India. Environmental Pollution. 213:338–46.
Ebadi, A., N.S.K. Sima, M. Olamaee, Hashemi, M. and Nasrabadi, R.G. 2018. Remediation of saline soils contaminated with crude oil using the halophyte Salicornia persica in conjunction with hydrocarbon-degrading bacteria. Journal of Environmental Management. 219:260-268.
El-Sheekh, M.M., Hamouda, R.A. and Niza, AA. (2013). Biodegradation of crude oil by Scenedesmus obliquus and Chlorella vulgaris growing under heterotrophic conditions. International Biodeterioration and Biodegradation. 82:67-72
EPA. (2005). Method 3535A. Solid–phase extraction (SPE), Revision1. Available t:http://www.epa.gov/osw/hazard/testmethods/sw846/online/3_series.htm
Eskandary, S., Tahmourespour, A., Hoodaji, M. and Abdillahi, A. (2017). The synergistic use of plant and isolated bacteria to clean up polycyclic aromatic hydrocarbons from contaminated soil. Journal of Environmental Health Science and Engineering. 15: 12-20.
Fatima, K., Imran, A., Naveed, M. and Afzal, M. (2017). Plant-bacteria synergism: An innovative approach for the remediation of crude oil- contaminated soils. Soil and Environment. 36(2): 93-113.
Guarino, C., D. Zuzolo, M. Marziano, B., Conte, G., Baiamonte, L., Morra, D., Benotti, D. Gresia, E. Robortella Stacul, Cicchella, D. and Sciarrillo, R. 2019. Investigation and Assessment for an effective approach to the reclamation of Polycyclic Aromatic Hydrocarbon (PAHs) contaminated site: SIN Bagnoli, Italy. Science Repository. 9: 11522-11534.
Han, G., B.X. Cui, Zhang, X.X. and Li, K.R. (2016). The effects of petroleum-contaminated soil on photosynthesis of Amorpha fruticosa seedlings. International Journal of Environmental Science Technology. 13:2383–2392
He, L.Y., Z.J. Chen, Ren, G.D. and Sheng, X.F. (2009). Increased cadmium and lead uptake of a cadmium hyperaccumulator tomato by cadmium-resistant bacteria. Ecotoxicology and Environmental Safety. 72: 1343-1348.
Huang, X.D., Y. El-Alawi, D.M. Penrose, Glick, B.R. and Greenberg, B.M. (2004). Responses of three grass species to creosote during phytoremediation. Environmental Pollution. 130: 453-463.
Javaheri, M., Mokhtati, M. and Samaei, M.R. (2019). Evaluation the Capability of Isolated Bacteria from Stabilized Compost for Bioremediation of Pyrene and Phenanthrene from Contaminated Soil with Municipal Solid Waste Leachate. Journal of Environmental Health and Sustainable Development. 4(3): 819-33.
Kalantary, R.R., A. Mohseni-Bandpi, A. Esrafili, S. Nasseri, F.R. Ashmagh, Jorfi, S. and Ja’fari, M. (2014). Effectiveness of biostimulation through nutrient content on the bioremediation of phenanthrene contaminated soil. Journal of Environmental Health Science and Engineering. 12(1):1-15.
Kosnar, Z. and Tlustos, P. (2018). Removal of soil polycyclic aromatic hydrocarbons derived from biomass fly ash by plants and organic amendments. Plant, Soil and Environment. 64 (2): 88-94.
Lichtenthaler, H.K. (1987). Chlorophylls and carotenoids, the pigments of photosynthetic biomembranes. Met. Enzymol. 148: 350-382
Lin, H., S. Tao, Zuo, Q. and Coveney, R. M. (2007). Uptake of polycyclic aromatic hydrocarbons by maize plants. Environmental Pollution. 148: 614–619.
Macci, C., S. Doni, E. Peruzzi, S. Bardella, G. Filippis, Ceccanti, B. and Masciandaro, G. (2012). A real-scale soil phytoremediation. Biodegradation. PubMed Retrieve. 24(4): 521-538.
Mishra, A. and Nautiyal, C. 2009. Functional diversity of the microbial community in the rhizosphere of chickpea grown in diesel fuelspiked soil amended with Trichoderma ressei using sole-carbon-source utilization profiles. World Journal of Microbiology and Biotechnology. 25: 1175–1180.
Mittal, A. and Singh, P. (2009). Isolation of hydrocarbon degrading bacteria from soils contaminated with crude oil spills. Indian Journal of Experimental Biology. 47: 760 -765
Motamedimehr, S.H. and Gitipour, S. (2019). Extraction and Recovery of Polycyclic Aromatic Hydrocarbons in Petroleum Contaminated Soils Using Supercritical Water by Response Surface Methodology. Pollution. 5(4): 913-922.
Mukred,A.M., A.A. Hamid, Hamzah, A. and Yusoff, W. 2008. Development of Three Bacteria Consortium for the Bioremediation of Crude Petroleum-oil in Contaminated Water. On Line Journal of Biological Sciences. 8 (4): 73-79.
Nnamchi, C.I., Obeta, J.A.N., and Ezeogu, L.I. 2006. Isolation and characterization of some poly aromatic hydrocarbon degrading bacteria from Nsukka soils in Nigeria. International Journal of Environmental Science and Technology. 3(2): 181-190
Oberai, M. and Khanna, V. (2018). Rhizoremediation – Plant Microbe Interactions in the Removal of Pollutants. International Journal of Current Microbiology and Applied Science. 7(1): 2280–2287.
Okoh, A.I. 2003. Biodegradation of Bonny light crude oil in soil microcosm by some bacterial strains isolated from crude oil flow stations saver pits in Nigeria. African Journal of Biotechnology. 2(5): 104-108
Polyak, Y.M., L.G. Bakina, M.V. Chugunova, Mayachkina, N.V. and Gerasimov Bure, A.O. (2018). Effect of remediation strategies on biological activity of oil-contaminated soil - A field study. International Biodeterioration and Biodegradation. 126: 57–68
Quinones-Aquilar, E. E., R. Ferra-Cerrato, R.F. Gavi, L. Fernandez, Rodriguez, V.R. and Alarcom, A. (2003). Emergence and growth of maize in a crude oil polluted soil. Agrociencia. 37: 585-594
Ravanipour, M., R.R. Kalantary, A. Mohseni-Bandpi, A. Esrafili, Farzadkia, S. and Hashemi-Najafabadi. M. (2015). Experimental design approach to the optimization of PAHs bioremediation from artificially contaminated soil: application of variables screening development. Journal of Environmental Health Science and Engineering. 13(1):1-10.
Seyed Alikhani, S., Shorafa, M., Tavasoli, A. and Ebrahimi Seyedeh, S. (2011). Effect of plants growth in different densities on the hydrocarbons of soil oil, soil and water. agricultural science and industries. 25:61-72.
Shim, H., S. Chauhan, D. Ryoo, K. Bowers, Thomas, S.M. and Burken, J.G. (2000). Rhizosphere competitiveness of trichloroethylene-degrading, poplar-colonizing recombinant bacteria. Appli Environmental Microbiology. 66(11): 4673-78
Smith, B., Stachowisk, M. and Volkenbugh, E. (1989). Cellular processes limiting leaf growth in plants under hypoxic root stress. Journal of Experimental Botany. 40(1):89-94
Swaathy, S, V., Kavitha., A.S., Pravin, Mandal A.B. and Gnanamani, A. (2014). Microbial surfactant mediated degradation of anthracene in aqueous phase by marine Bacillus licheniformis MTCC 5514. Biotechnology Reports. 4: 161–70
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Abena, M., T., Li, Shah, M. and Zhong, W. (2019). Biodegradation of total petroleum hydrocarbons (TPH) in highly contaminated soils by natural attenuation and bioaugmentation. Chemosphere. 234: 864-874.
Al-Hawas, G.H.S., W.M. Sukry, Azzoz, M.M. and Al-Moaik. R.M.S. (2012). The effect of sublethal concentrations of crude oil on the metabolism of Jojoba (Simmodsia chinensis) seedlings. International Research Journal of Plant Science. 3(4):54-62.
Bandowe, B. A. M., S. Leimer, Meusel, H. and Velescu, A. (2019). Plant diversity enhances the natural attenuation of polycyclic aromatic compounds (PAHs and oxygenated PAHs) in grassland soils. Soil Biology and Biochemistry. 129: 60-70.
Baruah, P., Saikia, R.R., Baruah, P.P. and Deka, S. (2014). Effect of crude oil contamination on the chlorophyll content and morpho-anatomy of Cyperus brevifolius (Rottb.). Environmental Science and Pollution Research. 21:12530–12538.
Bisht, S., Pandey, P., Bhargava, B.S., Sharma, Kumar, V. and Sharma, K.D. (2015). Bioremediation of polyaromatic hydrocarbons (PAHs) using rhizosphere technology. Brazilian Journal of Microbiology. 46: 7-21
Chiou, C., Sheng, G. and Manes, M. (2001). A partition-limited model for the plant uptake of organic contaminants from soil and water. Environmental Science and Technology. 35(7): 1437–44.
Chukwuma, C., Ikewuchi, J. and Monanu, M. (2019). Removal of hydrocarbons from crude oil contaminated agricultural soil by phytoremediation using Mariscus alternifolius and Fimbristylis ferruginea. European Journal of Biological Research. 9(1): 34-44.
D’Orazio, V., Ghanem, A. and Senesi, N. (2013). Phytoremediation of pyrene contaminated soils by different plant species. CLEAN–Soil. Air., Water. 41(4):377-82
Dasgupta, D., Jublee, J. and Suparna, M. (2018). Characterization, phylogenetic distribution and evolutionary trajectories of diverse hydrocarbon degrading microorganisms isolated from refinery sludge. Biotechnology. 8: 273-282.
Doyle, E., L. Muckian, Hickey, A.M. and Clipson, N. (2008). Microbial PAH degradation. Advances in Applied Microbiology. 65: 27–66
Dudhagara D.R., R.K. Rajpara, J.K., Bhatt, H.B., Gosai, Sachaniya, B.K. and Dave, B.P. (2016). Distribution, sources and ecological risk assessment of PAHs in historically contaminated surface sediments at Bhavnagar coast, Gujarat, India. Environmental Pollution. 213:338–46.
Ebadi, A., N.S.K. Sima, M. Olamaee, Hashemi, M. and Nasrabadi, R.G. 2018. Remediation of saline soils contaminated with crude oil using the halophyte Salicornia persica in conjunction with hydrocarbon-degrading bacteria. Journal of Environmental Management. 219:260-268.
El-Sheekh, M.M., Hamouda, R.A. and Niza, AA. (2013). Biodegradation of crude oil by Scenedesmus obliquus and Chlorella vulgaris growing under heterotrophic conditions. International Biodeterioration and Biodegradation. 82:67-72
EPA. (2005). Method 3535A. Solid–phase extraction (SPE), Revision1. Available t:http://www.epa.gov/osw/hazard/testmethods/sw846/online/3_series.htm
Eskandary, S., Tahmourespour, A., Hoodaji, M. and Abdillahi, A. (2017). The synergistic use of plant and isolated bacteria to clean up polycyclic aromatic hydrocarbons from contaminated soil. Journal of Environmental Health Science and Engineering. 15: 12-20.
Fatima, K., Imran, A., Naveed, M. and Afzal, M. (2017). Plant-bacteria synergism: An innovative approach for the remediation of crude oil- contaminated soils. Soil and Environment. 36(2): 93-113.
Guarino, C., D. Zuzolo, M. Marziano, B., Conte, G., Baiamonte, L., Morra, D., Benotti, D. Gresia, E. Robortella Stacul, Cicchella, D. and Sciarrillo, R. 2019. Investigation and Assessment for an effective approach to the reclamation of Polycyclic Aromatic Hydrocarbon (PAHs) contaminated site: SIN Bagnoli, Italy. Science Repository. 9: 11522-11534.
Han, G., B.X. Cui, Zhang, X.X. and Li, K.R. (2016). The effects of petroleum-contaminated soil on photosynthesis of Amorpha fruticosa seedlings. International Journal of Environmental Science Technology. 13:2383–2392
He, L.Y., Z.J. Chen, Ren, G.D. and Sheng, X.F. (2009). Increased cadmium and lead uptake of a cadmium hyperaccumulator tomato by cadmium-resistant bacteria. Ecotoxicology and Environmental Safety. 72: 1343-1348.
Huang, X.D., Y. El-Alawi, D.M. Penrose, Glick, B.R. and Greenberg, B.M. (2004). Responses of three grass species to creosote during phytoremediation. Environmental Pollution. 130: 453-463.
Javaheri, M., Mokhtati, M. and Samaei, M.R. (2019). Evaluation the Capability of Isolated Bacteria from Stabilized Compost for Bioremediation of Pyrene and Phenanthrene from Contaminated Soil with Municipal Solid Waste Leachate. Journal of Environmental Health and Sustainable Development. 4(3): 819-33.
Kalantary, R.R., A. Mohseni-Bandpi, A. Esrafili, S. Nasseri, F.R. Ashmagh, Jorfi, S. and Ja’fari, M. (2014). Effectiveness of biostimulation through nutrient content on the bioremediation of phenanthrene contaminated soil. Journal of Environmental Health Science and Engineering. 12(1):1-15.
Kosnar, Z. and Tlustos, P. (2018). Removal of soil polycyclic aromatic hydrocarbons derived from biomass fly ash by plants and organic amendments. Plant, Soil and Environment. 64 (2): 88-94.
Lichtenthaler, H.K. (1987). Chlorophylls and carotenoids, the pigments of photosynthetic biomembranes. Met. Enzymol. 148: 350-382
Lin, H., S. Tao, Zuo, Q. and Coveney, R. M. (2007). Uptake of polycyclic aromatic hydrocarbons by maize plants. Environmental Pollution. 148: 614–619.
Macci, C., S. Doni, E. Peruzzi, S. Bardella, G. Filippis, Ceccanti, B. and Masciandaro, G. (2012). A real-scale soil phytoremediation. Biodegradation. PubMed Retrieve. 24(4): 521-538.
Mishra, A. and Nautiyal, C. 2009. Functional diversity of the microbial community in the rhizosphere of chickpea grown in diesel fuelspiked soil amended with Trichoderma ressei using sole-carbon-source utilization profiles. World Journal of Microbiology and Biotechnology. 25: 1175–1180.
Mittal, A. and Singh, P. (2009). Isolation of hydrocarbon degrading bacteria from soils contaminated with crude oil spills. Indian Journal of Experimental Biology. 47: 760 -765
Motamedimehr, S.H. and Gitipour, S. (2019). Extraction and Recovery of Polycyclic Aromatic Hydrocarbons in Petroleum Contaminated Soils Using Supercritical Water by Response Surface Methodology. Pollution. 5(4): 913-922.
Mukred,A.M., A.A. Hamid, Hamzah, A. and Yusoff, W. 2008. Development of Three Bacteria Consortium for the Bioremediation of Crude Petroleum-oil in Contaminated Water. On Line Journal of Biological Sciences. 8 (4): 73-79.
Nnamchi, C.I., Obeta, J.A.N., and Ezeogu, L.I. 2006. Isolation and characterization of some poly aromatic hydrocarbon degrading bacteria from Nsukka soils in Nigeria. International Journal of Environmental Science and Technology. 3(2): 181-190
Oberai, M. and Khanna, V. (2018). Rhizoremediation – Plant Microbe Interactions in the Removal of Pollutants. International Journal of Current Microbiology and Applied Science. 7(1): 2280–2287.
Okoh, A.I. 2003. Biodegradation of Bonny light crude oil in soil microcosm by some bacterial strains isolated from crude oil flow stations saver pits in Nigeria. African Journal of Biotechnology. 2(5): 104-108
Polyak, Y.M., L.G. Bakina, M.V. Chugunova, Mayachkina, N.V. and Gerasimov Bure, A.O. (2018). Effect of remediation strategies on biological activity of oil-contaminated soil - A field study. International Biodeterioration and Biodegradation. 126: 57–68
Quinones-Aquilar, E. E., R. Ferra-Cerrato, R.F. Gavi, L. Fernandez, Rodriguez, V.R. and Alarcom, A. (2003). Emergence and growth of maize in a crude oil polluted soil. Agrociencia. 37: 585-594
Ravanipour, M., R.R. Kalantary, A. Mohseni-Bandpi, A. Esrafili, Farzadkia, S. and Hashemi-Najafabadi. M. (2015). Experimental design approach to the optimization of PAHs bioremediation from artificially contaminated soil: application of variables screening development. Journal of Environmental Health Science and Engineering. 13(1):1-10.
Seyed Alikhani, S., Shorafa, M., Tavasoli, A. and Ebrahimi Seyedeh, S. (2011). Effect of plants growth in different densities on the hydrocarbons of soil oil, soil and water. agricultural science and industries. 25:61-72.
Shim, H., S. Chauhan, D. Ryoo, K. Bowers, Thomas, S.M. and Burken, J.G. (2000). Rhizosphere competitiveness of trichloroethylene-degrading, poplar-colonizing recombinant bacteria. Appli Environmental Microbiology. 66(11): 4673-78
Smith, B., Stachowisk, M. and Volkenbugh, E. (1989). Cellular processes limiting leaf growth in plants under hypoxic root stress. Journal of Experimental Botany. 40(1):89-94
Swaathy, S, V., Kavitha., A.S., Pravin, Mandal A.B. and Gnanamani, A. (2014). Microbial surfactant mediated degradation of anthracene in aqueous phase by marine Bacillus licheniformis MTCC 5514. Biotechnology Reports. 4: 161–70