Isolation, identification and evaluation of oil hydrocarbon decomposing bacteria from contaminated areas of oil fields

Document Type: Research Paper


1 Department of Chemical Engineering, Qaemshahr Branch, Islamic Azad University, Qaemshahr, Iran.

2 Department of Chemical Engineering, Qaemshahr Branch, Islamic Azad University,


Microbialbiodegradation is known as an effective and harmless method to overcome environmental pollution with oil hydrocarbon. Some bacterial species were isolated from the Sarvestan oilfields (Iran, Fars province), then identified and applied for oil hydrocarbon decomposition. A carbon-free minimum medium (CFMM) containing 1% crude oil was used to isolate bacteria through incubation at 30°C in the dark at 200 rpm for 7 days. Different methods were used to identify the  hydrocarbon oil decomposing bacteria: gram staining, squalene hydrolysis, catalase, production of arginine dihydrolase, gelatin liquefaction, hydrogen sulfide production, levan production, methyl red, oxidase, nitrite reduction, oxidative/fermentative, starch hydrolysis and Tween-80 hydrolysis tests. Nine different oil decomposing bacterial species were isolated. All the species grew well at 28 and 35°C, while four isolates containing of Bacillus sp. SA13, Pantoea sp. SA1112, Pseudomonas aeruginosa sp. SA21, and Bacillus sp. SA23 were capable of growing in a temperature of up to around 42°C. The minimum salt tolerance for isolates, except for Enterobacter sp. SA711, was 8%; Bacillus sp. SA212 had the highest tolerance of 15% sodium chloride. Acinetobacter sp. SA172, Enterobacter sp. SA711, Pseudomonas sp. SA75, Bacillus sp. SA212 and Bacillus sp. SA23 had the most growth rate in the CFMM. The highest percentages of oil removal obtained were 89% for Enterobacter sp. SA711, 86% for Acinetobacter sp. SA172, and 68% for Pseudomonas sp. SA75. The three isolated bacterial strains from the contaminated soil of the Sarvestan area had a good ability to degrade oil hydrocarbon. Therefore, they could be used commercially for the bioremediation of this region.


Main Subjects

[1] Alikhani, J., Shayegan, J., Akbari, A. (2015). Risk assessment of hydrocarbon contaminant transport in vadose zone as it travels to groundwater table: A case study. Advances in environmental technology, 2, 77-84.

[2] Panda, S. K., Kar, R. N., Panda, C. R. (2013). Isolation and identification of petroleum hydrocarbon degrading microorganisms from oil contaminated environment. International journal of environmental sciences, 3(5), 1314-1321.

[3] Hazen, T. C., Prince, R. C., Mahmoudi, N. (2016). Marine oil biodegradation. Environmental science and technology, 50(2), 2121–2129

[4] Srivastava, J., Naraian, R., Kalra, S. J. S., Chandra, H. (2014). Advances in microbial bioremediation and the factors influencing the process. International Journal of environmental science and technology, 11(6), 1787-1800.

[5] Esmaeili, A. K. B. A. R., Sadeghi, E. (2014). The efficiency of Penicillium commune for bioremoval of industrial oil. International journal of environmental science and technology, 11(5), 1271-1276.

[6] Mani, D., Kumar, C. (2014). Biotechnological advances in bioremediation of heavy metals contaminated ecosystems: an overview with special reference to phytoremediation. International journal of environmental science and technology, 11(3), 843-872.

[7] Lai, C. C., Huang, Y. C., Wei, Y. H., Chang, J. S. (2009). Biosurfactant-enhanced removal of total petroleum hydrocarbons from contaminated soil. Journal of hazardous materials, 167(1-3), 609-614.

[8] Subathra, M. K., Immanuel, G., Suresh, A. H. (2013). Isolation and Identification of hydrocarbon degrading bacteria from Ennore creek. Bioinformation, 9(3), 150.

[9] Geetha, S. J., Joshi, S. J., Kathrotiya, S. (2013). Isolation and characterization of hydrocarbon degrading bacterial isolate from oil contaminated sites. APCBEE procedia, 5, 237-241.

[10]Rashedi, H. (2015). Indigenous production of biosurfactant and degradation of crude oil. Advances in environmental technology, 1(1), 11-16.

[11] Kostka, J. E., Prakash, O., Overholt, W. A., Green, S., Freyer, G., Canion, A., Huettel, M. (2011). Hydrocarbon-degrading bacteria and the bacterial community response in Gulf of Mexico beach sands impacted by the Deepwater Horizon oil spill. Applied and environmental microbiology, 77(22), 7962-7974.

[12] Liu, J., Bacosa, H. P., Liu, Z. (2017). Potential environmental factors affecting oil-degrading bacterial populations in deep and surface waters of the northern Gulf of Mexico. Frontiers in microbiology, 7, Article 2131, 1-14.

[13] Liu, Z., Liu, S. (2015). High phosphate concentrations accelerate bacterial peptide decomposition in hypoxic bottom waters of the northern Gulf of Mexico. Environmental science and technology, 50(2), 676-684.

[14] Schedler, M., Hiessl, R., Juárez, A. G. V., Gust, G., Müller, R. (2014). Effect of high pressure on hydrocarbon-degrading bacteria. AMB express, 4(1), 77-84.

[15] Bacosa, H. P., Liu, Z., Erdner, D. L. (2015). Natural sunlight shapes crude oil-degrading bacterial communities in Northern Gulf of Mexico surface waters. Frontiers in microbiology, 6, Article 1325, 1-14.

[16] Dubinsky, E. A., Conrad, M. E., Chakraborty, R., Bill, M., Borglin, S. E., Hollibaugh, J. T., Tom, L. M. (2013). Succession of hydrocarbon-degrading bacteria in the aftermath of the Deepwater Horizon oil spill in the Gulf of Mexico. Environmental science and technology, 47(19), 10860-10867.

[17] Shokrollahzadeh, S., Azizmohseni, F., Golmohamad, F. (2015). Characterization and kinetic study of PAH–degrading Sphingopyxis ummariensis bacteria isolated from a petrochemical wastewater treatment plant. Advances in environmental technology, 1(1), 1-9.

[18] Bao, M. T., Wang, L. N., Sun, P. Y., Cao, L. X., Zou, J., Li, Y. M. (2012). Biodegradation of crude oil using an efficient microbial consortium in a simulated marine environment. Marine pollution bulletin, 64(6), 1177-1185.

[19] Hassanshahian, M., Emtiazi, G., Cappello, S. (2012). Isolation and characterization of crude-oil-degrading bacteria from the Persian Gulf and the Caspian Sea. Marine pollution bulletin, 64(1), 7-12.

[20] Kurjogi, M. M., Kaliwal, B. B. (2018). Rapid and sensitive method for detection of Staphylococcus aureus enterotoxin genes in milk sample. Journal of applied biology and biotechnology Vol, 6(2), 15-19.

[21] Palacio-Bielsa, A., Pothier, J. F., Roselló, M., Duffy, B., López, M. M. (2012). Detection and identification methods and new tests as developed and used in the framework of COST 873 for bacteria pathogenic to stone fruits and nuts. Journal of plant pathology, 94(1sup), 1-135.

[22] Taylor, W. I., Achanzar, D. (1972). Catalase test as an aid to the identification of Enterobacteriaceae. Applied microbiology, 24(1), 58-61.

[23] Thirst, M. L. (1957). Gelatin liquefaction: a microtest. Journal of general microbiology, 17(2), 396-400.

[24] Clarke, P. H. (1953). Hydrogen sulphide production by bacteria. Journal of general microbiology, 8, 397-407.

[25] Pradhan, P. (2015). Tween 80 hydrolysis test. Mocrobiology and Infectious Diseases,

[26] Alegbeleye, O. O., Opeolu, B. O., Jackson, V. (2017). Bioremediation of polycyclic aromatic hydrocarbon (PAH) compounds:(acenaphthene and fluorene) in water using indigenous bacterial species isolated from the Diep and Plankenburg rivers, Western Cape, South Africa. Brazilian journal of microbiology, 48(2), 314-325.

[27] Parach, A., Rezvani, A., Assadi, M. M., Akbari-Adergani, B. (2017). Biodegradation of heavy crude oil using Persian Gulf autochthonous bacterium. International journal of environmental research, 11(5-6), 667-675.

[28] Ansari, N., Hassanshahian, M., Ravan, H. (2018). Study the Microbial Communities’ Changes in Desert and Farmland Soil After Crude Oil Pollution. International journal of environmental research, 12(3), 391-398.

[29] Toledo, F. L., Calvo, C., Rodelas, B., González-López, J. (2006). Selection and identification of bacteria isolated from waste crude oil with polycyclic aromatic hydrocarbons removal capacities. Systematic and applied microbiology, 29(3), 244-252.

[30] Hua, X., Wu, Z., Zhang, H., Lu, D., Wang, M., Liu, Y., Liu, Z. (2010). Degradation of hexadecane by Enterobacter cloacae strain TU that secretes an exopolysaccharide as a bioemulsifier. Chemosphere, 80(8), 951-956.

[31] Nkem, B. M., Halimoon, N., Yusoff, F. M., Johari, W. L. W., Zakaria, M. P., Medipally, S. R., Kannan, N. (2016). Isolation, identification and diesel-oil biodegradation capacities of indigenous hydrocarbon-degrading strains of Cellulosimicrobium cellulans and Acinetobacter baumannii from tarball at Terengganu beach, Malaysia.Marine pollution bulletin,107(1), 261-268.

[32] Shao, Y., Wang, Y., Wu, X., Xu, X., Kong, S., Tong, L., Li, B. (2015). Biodegradation of PAHs by Acinetobacter isolated from karst groundwater in a coal-mining area. Environmental earth sciences, 73(11), 7479-7488.

[33] Moscoso, F., Deive, F. J., Longo, M. A., Sanromán, M. A. (2015). Insights into polyaromatic hydrocarbon biodegradation by Pseudomonas stutzeri CECT 930: operation at bioreactor scale and metabolic pathways. International journal of environmental science and technology, 12(4), 1243-1252.

[34] Bisht, S., Pandey, P., Kaur, G., Aggarwal, H., Sood, A., Sharma, S, Bisht, N. S. (2014). Utilization of endophytic strain Bacillus sp. SBER3 for biodegradation of polyaromatic hydrocarbons (PAH) in soil model system. European journal of soil biology, 60, 67-76.

[35] Barin, R., Talebi, M., Biria, D., Beheshti, M. (2014). Fast bioremediation of petroleum-contaminated soils by a consortium of biosurfactant/bioemulsifier producing bacteria. International journal of environmental science and technology, 11(6), 1701-1710.

[36] Simard, R. G., Hasegawa, I., Bandaruk, W., Headington, C. E. (1951). Infrared spectrophotometric determination of oil and phenols in water. Analytical chemistry, 23(10), 1384-1387.

[37] Siddiqui, N., Rauf, A., Latif, A., Mahmood, Z. (2017). Spectrophotometric determination of the total phenolic content, spectral and fluorescence study of the herbal Unani drug Gul-e-Zoofa (Nepeta bracteata Benth). Journal of Taibah university medical sciences, 12(4), 360-363.