Main Article Content
Abstract
Soil contamination by petroleum hydrocarbons is a major problem resulting from activities related to petroleum industry, automobile service stations and accidental release of oil in the soil. Bioremediation/biodegradation can be considered as an effective aid to clean up oil spills, aside from conventional methods that are uneconomical and produce residues that harm the surrounding biota. There are an estimated over 250 mechanic shops with more than 5000 technicians in Maiduguri. Mechanics working therein frequently spill oil which is a potent source of immunotoxicants and carcinogenic to humans and animals. The aim of this study was to screen for petrophilic bacterial isolates from engine oil contaminated soil, that can utilize crude oil as sole source of carbon for growth in Maiduguri Metropolis. Ten isolates from engine oil contaminated soils in Bola and Tashan Kano areas of Borno State were obtained. The bacteria were individually cultured in Bushnell Haas Mineral Salt Medium devoid of carbon source except for crude oil for 21 days at 370C. Bacterial growth from visible increased turbidity was enumerated by CFU/g on nutrient agar. Pseudomonas sp. isolate exhibited relatively higher ability to grow on crude oil with TNTC followed by Bacillus sp., 6.7×104, Arthrobacter sp.,6.5×104, Flavobacterium sp.,6.2×104, Pseudomonas sp., 6×104, Nocardia sp.,5.9×104, Acinetobacter sp., 5×104, Bacillus sp., 5×104. Proteus sp., 4.7×104 and Flavobacterium sp. 4×104. It is therefore concluded that all the bacterial genera present in this study are effective hydrocarbon oil degraders.
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References
- Adebusoye, S. A., Amund, O. O., Ilori, M. O., Domeih, D. O., and Okpuzor, J. (2008). Growth and biosurfactant synthesis by Nigerian hydrocarbon-degrading estuarine bacteria. Revista de Biología Tropical, 56(4), 1603 611. doi:10.15517/rbt.v56i4.5746.
- Adsodun, J.K. and Mbagwu, J.S.C. (2008). Distribution of heavy metals and hydrocarbon contents in an alfisol contaminated with waste – lubricating oil amended with organic wastes. Bioresource Technology 99:3195-3204.
- Aina, O. R., Atuanya, E. I., Oshoma, C. E., Omotayo, A. E., and Olaleye, O. N. (2021). Biodegradation potential of rhizospheric microorganisms of rhizophora racemosa in crude oil contaminated mangrove swamp in the Niger delta. African Journal of Health, Safety and Environment, 2(2), 91-102.
- Ajona, M., and Vasanthi, P. (2021). Bioremediation of petroleum contaminated soils–A review. Materials Today: Proceedings 45(1).DOI: 10.1016/j.matpr.2021.01.949
- Aparna, C., Saitha, P., Himabindu, V., Alok, B., and Anjaneyulu, Y. (2010). Evaluation of bioremediation effectiveness on sediments contaminated with industrial wastes. International Journal of Environmental Science;1(4):607-620.
- Atlas, R. M. (1995). Hand Book of Microbiological Media. CRC press, London, pp:175.
- Azubuike, C. C., Chikere, C. B. and Okpokwasili, G. C. (2016). Bioremediation techniques – classification based on site of application: principles, advantages, limitations and prospects. World Journal of Microbiology and Biotechnology. 32 (11), 180.
- Chikere, C. B., Okpokwasili, G. C. and Chikere, B. O. (2012). Bioreactor-based bioremediation of hydrocarbon-polluted Niger Delta marine sediment, Nigeria. 3 Biotechnology, 53 66. http://doi.org/10.1007/s13205-011-0030-8.
- Chikere, C. B. and Ekwuabu, C. B. (2014). Molecular characterization of autochthonous hydrocarbon utilizing bacteria in oil polluted sites at Bodo community, Ogoni land, Niger Delta, Nigeria. Nig. Journal of Biotechnology; 27:28-33
- Colaninno, P. M. (2021). Identification of Gram-Positive Organisms. In Practical Handbook of Microbiology (pp. 51-58). CRC Press.
- Colborn, T., Vom Saal, F. S. and Solo, A. M. (1993). Developmental effects of endocrine-disrupting chemicals in wild life and humans. Environment Health Perspex.; 101(5): 378-84.
- Das, N. and Chandran, P. (2011). Microbial Degradation of Petroleum Hydrocarbon Contaminants: An Overview. Biotechnology Research International, 2011, Article ID:941810. http://dx.doi.org/10.4061/2011/941810.
- Delistraty, D. and Stone, A. (2007). Dioxins, metals, and fish toxicity in ash residue from space heaters burning used motor oil, Chemosphere, 68 (5), 907–914. doi:10.1016/j. chemosphere.2007.01.070
- Dominguez-Rosado, E. and Pichtel, J. (2004). Phytoremediation of soil contaminated with used motor oil: II. Greenhouse studies. Environmental Engineering Science, 21(2), 169-180.
- English, J. S. C., Dawe, R. S. and Ferguson, J. (2003). Environmental effects and skin disease. British Medical Bulletin, 68(1), 129-142.
- Eze, V. C., Okpokwasili, G. C. (2010). Microbial and other related changes in Niger Delta River sediment receiving industrial effluents. Continental Journal of Microbiology; 4:15-24.
- Okpokwasili, G.C. and Okorie, B. B. (1988). Biodeterioration potentials of microorganisms isolated from car engine lubricating oil. Tribology International: 21:4: 215 220. ISSN 0301 679X, https://doi.org/10.1016/0301-679x(88)90020-5.
- Gunasinghe, Y. H. K. I. S., Rathnayake, I. V. N. and Deeyamulla, M. P. (2021). Characterization of toluene degrading bacterial species isolated from soil. International Conference of the Biotechnology Society of Nepal (ICBSN). http://repository.kln.ac.lk/handle/123456789/24284
- Ibe, F. C., Duru, C. E., Isiuku, B. O. and Akalazu, J. N. (2021). Ecological risk assessment of the levels of polycyclic aromatic hydrocarbons in soils of the abandoned sections of Orji Mechanic Village, Owerri, Imo State, Nigeria. Bulletin of the National Research Centre, 45(1), 1-16.
- Ibiene, A. A., Orji, F. A. and Orji-Nwosu, E. C. (2011). Microbial population dynamics in crude oil-polluted soil in the Niger Delta. Nigerian Journal of Agriculture, Food and Environment; 7:8-13.
- Irwin, R. J., VanMouwerik, M., Stevens, L., Sees, M. D. and Basham, W. (1997). Environmental Contaminants Encyclopedia. National Park Service, Water ResourcesDivision, Fort Collins, Colorado.
- Jabir, D. M. and Lahmood, W. Y. (2021). Detection for enzymatic activity for bacteria isolated from the soil in polyethylene bags biodegradation. Materials Today: Proceedings.
- Kaszycki, P. P., Przemyslaw P. M. and Kolocze, H. (2011). Ex-Situ bioremediation of soil polluted with oily waste: The use of specialized microbial consortia for process bio augmentation. Ecological Chemistry and Engineering S:18:83-92.
- Kumar, M., Leon, V., Materano, A.D.S. and Ilzins, O.A.A. (2007). Halotolerant and thermotolerant Bacillus sp, degrades hydrocarbons and produces tension-active emulsifying agent. World Journal. Microbiol. Biotechnol ;23(2):211-220.
- Kuppusamy, S., Thavamani, P., Venka,T. K., Lee. Y.B., Naidu, R. and Megharaj, M. (2017). Remediation approaches for polycyclic aromatic hydrocarbons (PAHs) contaminated soils: technological constraints emerging trends and future directions. Chemosphere 168:944. http://doi.org/10.1016/j.chemoshere. 2016.10.115.
- Mandri, T. and Lin, J. (2007). Isolation and characterization of engine oil degrading indigenous microorganisms in Kwazulu-Natal, South Africa. African Journal of Biotechnology, 6(1):23-27.
- Mishra, S., Jyot, J., Kuhad, R. C. and Lal, B. (2001). Evaluation of inoculum addition to stimulate in situ bioremediation of oily sludge-contaminated soil. Applied Environmental Microbiology. 67, 1675-1681.
- Moyo, B. and Masika, P. J. (2009). Tick control methods used by resource-limited farmers and the effect of ticks on cattle in rural areas of the Eastern Cape Province, South Afriica. Tropical Animal Health and Production, 41(4), 517-523.
- Nathalia D. S., Alvarenga, Santos., Viniclus, R. R. and Marco Tulio, C. F. (2020). Review of engine journal bearing tribology in start-stop applications. http://doi.org/10.1016/j.engfailanal.2019.104344.
- Nivedita, P. and Krushna, P. S. (2020). Characterization of waste engine oil (WEO) pyrolytic oil and diesel blended oil. Fuel Properties and Compositional Analysis. http://doi.org/10.1016/j.matpr.2020.02.679.
- Nweke, C. O. and Okpokwasili, G. C.(2004). Effects of bioremediation treatments on the bacterial populations of soil at different depths. Nigeria Journal of Microbiology: 18: 362-372.
- Obire, O. and Nwanbeta, O. (2002). Effects of refined petroleum hydrocarbon on soil physiochemical and bacteriological characteristics. Journal of Applied Science and Environmental Management: 6 (1):34-44.
- Okerentugba, P. O. and Ezeronye, O. U. (2003). Petroleum Degrading Potentials of Single and Mixed Microbial Cultures Isolated from Rivers and Refinery Effluents in Nigeria. African Journal of Biotechnology: 2 (9): 293-295.
- Oko-Oboh, E., Oviasogie, P. O., Senjodi, B.A. and Oriafo, S. (2016). Characterization of alluvial soils in a derived savannah ecology in Edo State Southern Nigeria. Nigerian Journal of Agriculture, Food and Environment:12(3):204-209.
- Okonokhu, B. O., Ikhajagbe, B., Anoliefo, G. and Emede, T. (2007). The effects of Spent Engine Oil on Soil Properties and Growth of Maize (Zea mays L.). Journal of Applied Sciences and Environmental Management (ISSN:1119-8362) Vol 11Num 3.11.10. 4314/jasem. V11i3. 55162.
- Onwa Ndubuisi, C., Onochie, C. C. and Nwadiogbu, I. (2018). Isolation of hydrocarbon degrading bacteria species from soil contaminated with lubricating oil in Enugu South East Nigeria. International Digital Organisation for Scientific Research (idosr jounal of applied sciences): 3(1):76-85.
- Onwurah, I. N. E., Ogugua, V. N., Onyike, N. B. Ochonogor, A. E. and Otitoju, O. F., (2003). Crude Oil Spills in the Environment, Effects and some Innovative Clean-up Biotechnologies. International Journal of Environmental Research, 1(4), pp: 307-320.
- Perez-Cadahia, B., Lafuente, A., Cabaleiro, T., Pasaro, E., Mendez, J. and Laffon, B. (2007). Initial Study on the effects of Prestige oil on human health. EnvironmentInternational; 33:176-185.
- Prince, R. C. (1993). Petroleum spill bioremediation in marine experiments. Microbiology Review. 19:217-242.
- Prenafeta-Boldu, X. F., Kuhn, A., Dmam, L., Anke, H. and Bont, J. D. (2001). Isolation and characterization of fungi growing on volatile aromatic hydrocarbons as their sole carbon and energy source. Mycology Research. 4:477-484.
- Alvarez, P. J. J. and Vogel, T. M. (1991). Substrate interaction of benzene, toluene, and paraxylene during microbial degradation by pure cultures and mixed culture aquifer slurries. Appliedand Environmental Microbiology: 57(10): 2981-2985.
- Pritchard, P.H., Mueller, J.G. and Rogers, J.C. (1992). Oil spill bioremediation: experiences, lessons and results from the Exxon Valdez oil spill in Alaska. Biodegradation: 3: 315 335:https://doi.org/10.1007/BF00129091.
- Propst, T, L., Lochmiller, R. L., Qualis, C. W. Jr. and McBee, K. (1999). In situ (mesocosm) assessment of immunotoxicity risks to small mammals inhabiting petrochemical waste sites. Chemosphere: 38: 1049-1067.
- Rahman, K. S., Rahman T., Laskshman, P. and Banat, I. (2002). Occurrence of crude oil degrading bacteria in gasoline and diesel station soils. Journal of Basic Microbiology, 42: 284-291.
- Roling, W. F. M., Milner, M. G., Jones, D. M., Lee, K., Daniel, F., Swannell, R. J. P. and Head, I. M. (2002). Robust hydrocarbon degradation and hydrocarbon degradation and dynamics of bacterial communities during nutrient-enhanced oil spill bioremediation. Applied Environmental. Microbiology, 68(11):5537-5548.
- Sarma, A. and Sarma, H. (2010). Enhanced biodegradation by some microbial isolates supplemented with heavy metals. International Journal of Biotechnology: 6:441-448.
- Sepahi, A. A., Golpasha, I. D., Emanmi, M. and Nakhoda, A. M. (2008). Isolation and characterization of crude oil degrading Bacillus spp. Iran Journal of Environmental Health Science and Engineering: 5(3):149-154
- Speight, J.G (1991). The chemistry and technology of petroleum. Lewis Publishers, Marcel Dekker, New York, pp.30
- Stegmann, R., Lolter, S. and Heerenklage, J. (2011). Biological treatment of oil-contaminated soils in bioreactors. International Journal of Microbiology: 4(3):231-235.
- Udeani, T. K. C., Obroh, A. A., Okwuosa, C. N., Achukwu, P. U., and Azubike, N. (2009). Isolation of bacteria from mechanic workshops’ soil environment contaminated with used engine oil. African journal of Biotechnology: 8: 22
- Vilayutham, T., Arutchelvan, V., Nagarajan, S. and Muralikrishnan, V. (2012). Isolation and identification of polycyclic aromatic hydrocarbon-degrading bacteria from crude oil exploration bore well sludge. BioremediationJournal:16(3):141-146.
- Vitalina, L., Oleksandr, T. and Yevheniia, A. (2020). Environmental safety of motor transport enterprises within the urban areas. Journal of Ecological Engineering: 21: (4), 2020.
References
Adebusoye, S. A., Amund, O. O., Ilori, M. O., Domeih, D. O., and Okpuzor, J. (2008). Growth and biosurfactant synthesis by Nigerian hydrocarbon-degrading estuarine bacteria. Revista de Biología Tropical, 56(4), 1603 611. doi:10.15517/rbt.v56i4.5746.
Adsodun, J.K. and Mbagwu, J.S.C. (2008). Distribution of heavy metals and hydrocarbon contents in an alfisol contaminated with waste – lubricating oil amended with organic wastes. Bioresource Technology 99:3195-3204.
Aina, O. R., Atuanya, E. I., Oshoma, C. E., Omotayo, A. E., and Olaleye, O. N. (2021). Biodegradation potential of rhizospheric microorganisms of rhizophora racemosa in crude oil contaminated mangrove swamp in the Niger delta. African Journal of Health, Safety and Environment, 2(2), 91-102.
Ajona, M., and Vasanthi, P. (2021). Bioremediation of petroleum contaminated soils–A review. Materials Today: Proceedings 45(1).DOI: 10.1016/j.matpr.2021.01.949
Aparna, C., Saitha, P., Himabindu, V., Alok, B., and Anjaneyulu, Y. (2010). Evaluation of bioremediation effectiveness on sediments contaminated with industrial wastes. International Journal of Environmental Science;1(4):607-620.
Atlas, R. M. (1995). Hand Book of Microbiological Media. CRC press, London, pp:175.
Azubuike, C. C., Chikere, C. B. and Okpokwasili, G. C. (2016). Bioremediation techniques – classification based on site of application: principles, advantages, limitations and prospects. World Journal of Microbiology and Biotechnology. 32 (11), 180.
Chikere, C. B., Okpokwasili, G. C. and Chikere, B. O. (2012). Bioreactor-based bioremediation of hydrocarbon-polluted Niger Delta marine sediment, Nigeria. 3 Biotechnology, 53 66. http://doi.org/10.1007/s13205-011-0030-8.
Chikere, C. B. and Ekwuabu, C. B. (2014). Molecular characterization of autochthonous hydrocarbon utilizing bacteria in oil polluted sites at Bodo community, Ogoni land, Niger Delta, Nigeria. Nig. Journal of Biotechnology; 27:28-33
Colaninno, P. M. (2021). Identification of Gram-Positive Organisms. In Practical Handbook of Microbiology (pp. 51-58). CRC Press.
Colborn, T., Vom Saal, F. S. and Solo, A. M. (1993). Developmental effects of endocrine-disrupting chemicals in wild life and humans. Environment Health Perspex.; 101(5): 378-84.
Das, N. and Chandran, P. (2011). Microbial Degradation of Petroleum Hydrocarbon Contaminants: An Overview. Biotechnology Research International, 2011, Article ID:941810. http://dx.doi.org/10.4061/2011/941810.
Delistraty, D. and Stone, A. (2007). Dioxins, metals, and fish toxicity in ash residue from space heaters burning used motor oil, Chemosphere, 68 (5), 907–914. doi:10.1016/j. chemosphere.2007.01.070
Dominguez-Rosado, E. and Pichtel, J. (2004). Phytoremediation of soil contaminated with used motor oil: II. Greenhouse studies. Environmental Engineering Science, 21(2), 169-180.
English, J. S. C., Dawe, R. S. and Ferguson, J. (2003). Environmental effects and skin disease. British Medical Bulletin, 68(1), 129-142.
Eze, V. C., Okpokwasili, G. C. (2010). Microbial and other related changes in Niger Delta River sediment receiving industrial effluents. Continental Journal of Microbiology; 4:15-24.
Okpokwasili, G.C. and Okorie, B. B. (1988). Biodeterioration potentials of microorganisms isolated from car engine lubricating oil. Tribology International: 21:4: 215 220. ISSN 0301 679X, https://doi.org/10.1016/0301-679x(88)90020-5.
Gunasinghe, Y. H. K. I. S., Rathnayake, I. V. N. and Deeyamulla, M. P. (2021). Characterization of toluene degrading bacterial species isolated from soil. International Conference of the Biotechnology Society of Nepal (ICBSN). http://repository.kln.ac.lk/handle/123456789/24284
Ibe, F. C., Duru, C. E., Isiuku, B. O. and Akalazu, J. N. (2021). Ecological risk assessment of the levels of polycyclic aromatic hydrocarbons in soils of the abandoned sections of Orji Mechanic Village, Owerri, Imo State, Nigeria. Bulletin of the National Research Centre, 45(1), 1-16.
Ibiene, A. A., Orji, F. A. and Orji-Nwosu, E. C. (2011). Microbial population dynamics in crude oil-polluted soil in the Niger Delta. Nigerian Journal of Agriculture, Food and Environment; 7:8-13.
Irwin, R. J., VanMouwerik, M., Stevens, L., Sees, M. D. and Basham, W. (1997). Environmental Contaminants Encyclopedia. National Park Service, Water ResourcesDivision, Fort Collins, Colorado.
Jabir, D. M. and Lahmood, W. Y. (2021). Detection for enzymatic activity for bacteria isolated from the soil in polyethylene bags biodegradation. Materials Today: Proceedings.
Kaszycki, P. P., Przemyslaw P. M. and Kolocze, H. (2011). Ex-Situ bioremediation of soil polluted with oily waste: The use of specialized microbial consortia for process bio augmentation. Ecological Chemistry and Engineering S:18:83-92.
Kumar, M., Leon, V., Materano, A.D.S. and Ilzins, O.A.A. (2007). Halotolerant and thermotolerant Bacillus sp, degrades hydrocarbons and produces tension-active emulsifying agent. World Journal. Microbiol. Biotechnol ;23(2):211-220.
Kuppusamy, S., Thavamani, P., Venka,T. K., Lee. Y.B., Naidu, R. and Megharaj, M. (2017). Remediation approaches for polycyclic aromatic hydrocarbons (PAHs) contaminated soils: technological constraints emerging trends and future directions. Chemosphere 168:944. http://doi.org/10.1016/j.chemoshere. 2016.10.115.
Mandri, T. and Lin, J. (2007). Isolation and characterization of engine oil degrading indigenous microorganisms in Kwazulu-Natal, South Africa. African Journal of Biotechnology, 6(1):23-27.
Mishra, S., Jyot, J., Kuhad, R. C. and Lal, B. (2001). Evaluation of inoculum addition to stimulate in situ bioremediation of oily sludge-contaminated soil. Applied Environmental Microbiology. 67, 1675-1681.
Moyo, B. and Masika, P. J. (2009). Tick control methods used by resource-limited farmers and the effect of ticks on cattle in rural areas of the Eastern Cape Province, South Afriica. Tropical Animal Health and Production, 41(4), 517-523.
Nathalia D. S., Alvarenga, Santos., Viniclus, R. R. and Marco Tulio, C. F. (2020). Review of engine journal bearing tribology in start-stop applications. http://doi.org/10.1016/j.engfailanal.2019.104344.
Nivedita, P. and Krushna, P. S. (2020). Characterization of waste engine oil (WEO) pyrolytic oil and diesel blended oil. Fuel Properties and Compositional Analysis. http://doi.org/10.1016/j.matpr.2020.02.679.
Nweke, C. O. and Okpokwasili, G. C.(2004). Effects of bioremediation treatments on the bacterial populations of soil at different depths. Nigeria Journal of Microbiology: 18: 362-372.
Obire, O. and Nwanbeta, O. (2002). Effects of refined petroleum hydrocarbon on soil physiochemical and bacteriological characteristics. Journal of Applied Science and Environmental Management: 6 (1):34-44.
Okerentugba, P. O. and Ezeronye, O. U. (2003). Petroleum Degrading Potentials of Single and Mixed Microbial Cultures Isolated from Rivers and Refinery Effluents in Nigeria. African Journal of Biotechnology: 2 (9): 293-295.
Oko-Oboh, E., Oviasogie, P. O., Senjodi, B.A. and Oriafo, S. (2016). Characterization of alluvial soils in a derived savannah ecology in Edo State Southern Nigeria. Nigerian Journal of Agriculture, Food and Environment:12(3):204-209.
Okonokhu, B. O., Ikhajagbe, B., Anoliefo, G. and Emede, T. (2007). The effects of Spent Engine Oil on Soil Properties and Growth of Maize (Zea mays L.). Journal of Applied Sciences and Environmental Management (ISSN:1119-8362) Vol 11Num 3.11.10. 4314/jasem. V11i3. 55162.
Onwa Ndubuisi, C., Onochie, C. C. and Nwadiogbu, I. (2018). Isolation of hydrocarbon degrading bacteria species from soil contaminated with lubricating oil in Enugu South East Nigeria. International Digital Organisation for Scientific Research (idosr jounal of applied sciences): 3(1):76-85.
Onwurah, I. N. E., Ogugua, V. N., Onyike, N. B. Ochonogor, A. E. and Otitoju, O. F., (2003). Crude Oil Spills in the Environment, Effects and some Innovative Clean-up Biotechnologies. International Journal of Environmental Research, 1(4), pp: 307-320.
Perez-Cadahia, B., Lafuente, A., Cabaleiro, T., Pasaro, E., Mendez, J. and Laffon, B. (2007). Initial Study on the effects of Prestige oil on human health. EnvironmentInternational; 33:176-185.
Prince, R. C. (1993). Petroleum spill bioremediation in marine experiments. Microbiology Review. 19:217-242.
Prenafeta-Boldu, X. F., Kuhn, A., Dmam, L., Anke, H. and Bont, J. D. (2001). Isolation and characterization of fungi growing on volatile aromatic hydrocarbons as their sole carbon and energy source. Mycology Research. 4:477-484.
Alvarez, P. J. J. and Vogel, T. M. (1991). Substrate interaction of benzene, toluene, and paraxylene during microbial degradation by pure cultures and mixed culture aquifer slurries. Appliedand Environmental Microbiology: 57(10): 2981-2985.
Pritchard, P.H., Mueller, J.G. and Rogers, J.C. (1992). Oil spill bioremediation: experiences, lessons and results from the Exxon Valdez oil spill in Alaska. Biodegradation: 3: 315 335:https://doi.org/10.1007/BF00129091.
Propst, T, L., Lochmiller, R. L., Qualis, C. W. Jr. and McBee, K. (1999). In situ (mesocosm) assessment of immunotoxicity risks to small mammals inhabiting petrochemical waste sites. Chemosphere: 38: 1049-1067.
Rahman, K. S., Rahman T., Laskshman, P. and Banat, I. (2002). Occurrence of crude oil degrading bacteria in gasoline and diesel station soils. Journal of Basic Microbiology, 42: 284-291.
Roling, W. F. M., Milner, M. G., Jones, D. M., Lee, K., Daniel, F., Swannell, R. J. P. and Head, I. M. (2002). Robust hydrocarbon degradation and hydrocarbon degradation and dynamics of bacterial communities during nutrient-enhanced oil spill bioremediation. Applied Environmental. Microbiology, 68(11):5537-5548.
Sarma, A. and Sarma, H. (2010). Enhanced biodegradation by some microbial isolates supplemented with heavy metals. International Journal of Biotechnology: 6:441-448.
Sepahi, A. A., Golpasha, I. D., Emanmi, M. and Nakhoda, A. M. (2008). Isolation and characterization of crude oil degrading Bacillus spp. Iran Journal of Environmental Health Science and Engineering: 5(3):149-154
Speight, J.G (1991). The chemistry and technology of petroleum. Lewis Publishers, Marcel Dekker, New York, pp.30
Stegmann, R., Lolter, S. and Heerenklage, J. (2011). Biological treatment of oil-contaminated soils in bioreactors. International Journal of Microbiology: 4(3):231-235.
Udeani, T. K. C., Obroh, A. A., Okwuosa, C. N., Achukwu, P. U., and Azubike, N. (2009). Isolation of bacteria from mechanic workshops’ soil environment contaminated with used engine oil. African journal of Biotechnology: 8: 22
Vilayutham, T., Arutchelvan, V., Nagarajan, S. and Muralikrishnan, V. (2012). Isolation and identification of polycyclic aromatic hydrocarbon-degrading bacteria from crude oil exploration bore well sludge. BioremediationJournal:16(3):141-146.
Vitalina, L., Oleksandr, T. and Yevheniia, A. (2020). Environmental safety of motor transport enterprises within the urban areas. Journal of Ecological Engineering: 21: (4), 2020.