Environmental Science and Pollution Research

, Volume 21, Issue 18, pp 10725–10735 | Cite as

Assessing the hydrocarbon degrading potential of indigenous bacteria isolated from crude oil tank bottom sludge and hydrocarbon-contaminated soil of Azzawiya oil refinery, Libya

  • Abdulatif A. Mansur
  • Eric M. Adetutu
  • Krishna K. Kadali
  • Paul D. Morrison
  • Yuana Nurulita
  • Andrew S. Ball
Research Article


The disposal of hazardous crude oil tank bottom sludge (COTBS) represents a significant waste management burden for South Mediterranean countries. Currently, the application of biological systems (bioremediation) for the treatment of COTBS is not widely practiced in these countries. Therefore, this study aims to develop the potential for bioremediation in this region through assessment of the abilities of indigenous hydrocarbonoclastic microorganisms from Libyan Hamada COTBS for the biotreatment of Libyan COTBS-contaminated environments. Bacteria were isolated from COTBS, COTBS-contaminated soil, treated COTBS-contaminated soil, and uncontaminated soil using Bushnell Hass medium amended with Hamada crude oil (1 %) as the main carbon source. Overall, 49 bacterial phenotypes were detected, and their individual abilities to degrade Hamada crude and selected COBTS fractions (naphthalene, phenanthrene, eicosane, octadecane and hexane) were evaluated using MT2 Biolog plates. Analyses using average well colour development showed that ~90 % of bacterial isolates were capable of utilizing representative aromatic fractions compared to 51 % utilization of representative aliphatics. Interestingly, more hydrocarbonoclastic isolates were obtained from treated contaminated soils (42.9 %) than from COTBS (26.5 %) or COTBS-contaminated (30.6 %) and control (0 %) soils. Hierarchical cluster analysis (HCA) separated the isolates into two clusters with microorganisms in cluster 2 being 1.7- to 5-fold better at hydrocarbon degradation than those in cluster 1. Cluster 2 isolates belonged to the putative hydrocarbon-degrading genera; Pseudomonas, Bacillus, Arthrobacter and Brevundimonas with 57 % of these isolates being obtained from treated COTBS-contaminated soil. Overall, this study demonstrates that the potential for PAH degradation exists for the bioremediation of Hamada COTBS-contaminated environments in Libya. This represents the first report on the isolation of hydrocarbonoclastic bacteria from Libyan COTBS and COTBS-contaminated soil.


Weathered Hamada crude oil Crude oil tank bottom sludge (COTBS) Contaminated soil MT2 Biolog plates Hydrocarbonoclastic bacteria Bioremediation 



This work was supported by the Libyan Ministry of Higher Education and Science Research. The authors thank the Environmental and Natural Resources Engineering, Azzawiya University, Libya, the management of Azzawiya oil refinery, Libya, and brothers Ali Mansour, Mohamad Ahfeda and Nassradden Souf for supplying soil and COTBS samples.

Supplementary material

11356_2014_3018_MOESM1_ESM.pptx (364 kb)
Supplementary Fig. 1 Figure. 1a Chromatogram of soil extract obtained from treated COTBS contaminated soil. Figure. 1 b: Chromatogram of COTBS. (PPTX 363 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Abdulatif A. Mansur
    • 1
    • 2
  • Eric M. Adetutu
    • 1
  • Krishna K. Kadali
    • 1
  • Paul D. Morrison
    • 1
  • Yuana Nurulita
    • 1
    • 3
  • Andrew S. Ball
    • 1
  1. 1.School of Applied ScienceRoyal Melbourne Institute of TechnologyMelbourneAustralia
  2. 2.Environmental and Natural Resources Engineering, Faculty of EngineeringAzawia UniversityAzawiaLibya
  3. 3.Chemistry Department, Faculty of Mathematics and Natural SciencesUniversity of RiauPekanbaruIndonesia

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