Isolation and molecular identification of hydrocarbon-degrading bacteria from soil around Tabriz refinery

Document Type : Original Article


1 Department of Natural Resources, Isfahan University of Technology, Isfahan, Iran

2 Department of Biology, Faculty of Sciences, Isfahan University of Falavajan, Isfahan, Iran


A wide range of organic chemicals enter into the environment, intentionally or unintentionally, creating public concern. Petroleum hydrocarbons represent one of these organic pollutants threatening various hazards to terrestrial and marine ecosystems. We need cost effective technologies for restoration of contaminated sites, and bioremediation is one the methods which has been used in recent decades. In this method, microrganisms and plants are used to remove, degrade or stabilize pollutants. Biological methods are economically more acceptable than physical and chemical methods. The current study aims to identify the best bacteria for decomposing petroleum hydrocarbons in the soil at Tabriz refinery.
Material and methods:
In this study, oil contaminated soils were collected from a refinery in Tabriz and, in order to identify them, a synthetic Bushnell Haas Mineral Salts medium (BHMS) was used. After the enrichment process, separation and purification were performed. Two strains were selected for molecular identification based on the higher growth in a medium containing crude oil. Molecular identification of the selected strains was conducted after DNA extraction and sequencing of the 16S rRNA gene. The growth of bacteria in both light and heavy diesel was recorded using a spectrophotometer at wavelengths of 560 nm. To assess the production of a biosurfactant, several tests including a drop collapsing test, oil displacement test as well as emulsification activity and cell surface hydrophobicity were performed.
Results and discussion:
The sequences obtained were compared with those of the National Center for Biotechnology Information (NCBI). The isolated bacteria with 98% similarity belonged to Pseudomonas aeruginosa and Achromobacter spanius. Both bacteria showed growth in all the three oil components, however,Achromobacter spanius had higher growth rate compared to Pseudomonas aeruginosa. The latter could decompose 67 percent of light diesel in 9 days. Although both bacteria could produce a biosurfactant, Achromobacter spanius had higher hydrophobic cell surface and emulsification activity.
In the current study we introduced Achromobacter spanius as an efficient bacterium for the decomposition of heavy and light diesel and crude oil. Based on the results, it is recomended that the ability of the identified bacteria in petroleum contaminated terrestrial and aquatic environments be investigated further.


  1. Adli, M., Hassanshahian, M. and Krimi Niki, A., 2013. Isolation, identification and characterization of two species of Shyvanlay biosurfactantproducing Persian Gulf. Joranl. Microbes World. 6, 53-61. ( In Persian with English abstract).
  2. Cheng Deng, M., Li, J., Rui Liang, F., Yi, M., Ming Xu, X., Ping Yuan, J., Peng, J. and Fei Wu, C., 2014. Isolation and characterization of a novel hydrocarbon-degrading bacterium Achromobacter sp. HZ01 from the crude oil-contaminated seawater at the Daya Bay, southern China. Marine Pollution Journal. 83, 79-86.
  3. Crapez, M. and Wasserman, J., 2006. Characterization of hydrocarbonoclastic bacterial communities from mangrove sediments in Guanabara Bay, Brazil. Research in Microbiology Jornal. 157, 752–762.
  4. CapelliS, M., Busalmen, J. and Sanchez, S., 2001. “Hydrocarbon bioremediation of a mineral-base contaminated waste from crude oil extraction by Indiagenous bacteria”. International Biodeterioration and Biodegradation.. 4, 233-23.
  5. Hasanshahian, M. and Emtiazi , G., 2008. Investigation of alkane biodegradation using the microtiter plate method and correlation between biofilm formation, biosurfactant production and crude oil biodegradation. International Biodeterioration and Biodegradation Jornal. 62, 170–178.
  6. Hassanshahian, M., Emtiazi, G. and Cappello , S., 2012. Isolation and characterization of crudeoil-degrading bacteria from the Persian Gulf and the Caspian Sea. Marine Pollution Bulletin Jornal. 64, 7-12.
  7. Hassanshahian, M., Ahmadinejad, M. Tebyanian, H. and Kariminik , A., 2013. Isolation and chracterization of alkan degrding bacteria from peteroleum reservoir waste water in Iran (Kerman and Tehran provinances ). Marin Pollution Bulletine Jornal. 73, 300-305.
  8. Hassanshahian, M., Zeynalipour, M. and Musa, F., 2014. Isolation and characterization of crude oil degrading bacteria from the Persian Gulf (Khorramshahr provinnce). Marine Pollution Bulletin Jornal. 82, 39-44.
  9. Hassanshahian, M., 2013. Isolation and characterization of biosurfactant producing bacteria from Persian Gulf (Bushehr provenance). Marine Pollution Bulletin Jornal. 86, 361-366.
  10. Isaac , P., Sanchez , L., Bourguignon, N., Eugenia Cabral, M. and Ferrero, M., 2013. Indigenous PAH-degrading bacteria from oil-polluted sediments in Caleta Cordova, Patagonia Argentina. International Biodeterioration and Biodegradation Jornal. 82, 207-214
  11. Kumaria, B., Singha ,S. and Singhb, P., 2012. Characterization of two biosurfactant producing strains in crude oil degradation. Process Biochemistry Journal. 47, 2463–2471.
  12. Liu, H., Yao, J., Yuan, Z., Shang, Y , Chen, H., Wang, F. and Masakorala , K., 2014. Isolation and characterization of crude oil degrading bacteria from oil water mixture in Dagang oil field China. International Biodeterioration and Biodegradation Jornal. 87, 52-59.
  13. Margareth, E., Guyoneaud, R., Marisol GoñiUrriza, M., Ranchou-Peyruse, A., Verbaere, A., Crapez, M . and Wasserman, J., 2006. Characterization of hydrocarbonoclastic bacterial communities from mangrove sediments in Guanabara Bay, Brazil. Research in Microbiology Jornal. 157, 752–762.
  14. Rosanohernandez , M., Sadd , H. and linares, L., 2012. Petroleum-influenced beach sediment of the cameche bank mexico: Diversityand bacterial community structure assessment. Journal of Environmental anagement Journal.
  15. Silva, T., Verde, L., Santos Neto, E. and Oliveira, V., 2013. Diversity analyses of microbial communities in petroleum sample from Brazilian oil field. International Biodeterioration and Biodegradation Jornal. 81, 57-70.
  16. Soleimani , M., Farhoudi, M. and Christensen, J., 2013. Chemometric assessment of enhanced bioremediation of oil contaminated soils. Journal of Hazardous Materials Jornal. 254, 372– 381.
  17. Shah alyan , F., Safahiyh, A., salamat, N., Mojodi, F. and Zareh Dost, M., 2014. Check growth and production of biosurfactants two species Pseudomonas aeruginosa, Pseudomonas putida in Moses cove. Journal of Aquatic Ecology. 5, 51- 59.(In Persian with English abstract).
  18. Tebyanian , H. Hassanshahian , M. Kariminik, A.2013. Isolation and characterization of alkane degrading bacteria from petroleum reservoir waste water in Iran (Kerman and Tehran provenances). Journalm of Microbes World. 4, 105-114. (In Persian with English abstract).
  19. Vidali, M., 2001. Bioremediation. An overview. Pure and Applied Chemistry Jornal. 73, 1163– 1172.
  20. Wilson , S. and Jones, K., 1993. Bioremedation of soil contaminated with polynuclear aromatic hydrocarbons (PAH). Environmental Polluation Jornal. 81, 229-249.