One of the major environmental problems today is hydrocarbon contamination resulting from the activities related to the petrochemical industry. Accidental releases of petroleum products are of particular concern in the environment. Hydrocarbon components have been known to belong to the family of carcinogens and neurotoxic organic pollutants. Currently accepted disposal methods of incineration or burial insecure landfills can become prohibitively expensive when amounts of contaminants are large. Mechanical and chemical methods generally used to remove hydrocarbons from contaminated sites have limited effectiveness and can be expensive. Bioremediation is the promising technology for the treatment of these contaminated sites since it is cost-effective and will lead to complete mineralization. Bioremediation functions basically on biodegradation, which may refer to complete mineralization of organic contaminants into carbon dioxide, water, inorganic compounds, and cell protein or transformation of complex organic contaminants to other simpler organic compounds by biological agents like microorganisms. Many indigenous microorganisms in water and soil are capable of degrading hydrocarbon contaminants. This paper presents an updated overview of petroleum hydrocarbon degradation by microorganisms under different ecosystems.

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Five yeast species, namely Candida tropicalis, Cryptococcus laurentii, Trichosporon asahii, Rhodotorula mucilaginosa and Candida rugosa isolated from hydrocarbon-contaminated soil were found to be potent degraders of diesel oil. These microorganisms showed the presence of enzymes cytochrome P450, NADPH cytochrome c reductase, aminopyrine N demethylase, alcohol dehydrogenase, aldehyde dehydrogenase, naphthalene dioxygenase, catalase and glutathione S transferase when the cells were incubated for 48 h in Bushnell Haas medium supplemented with 2% diesel oil as the sole source of carbon. The cytochrome P450 monooxygenase enzyme system was found to play an important role in diesel oil degradation. A plasmid approximately 12kb in size was found to be harboured by all the yeast species. The role of the plasmid on diesel oil degradation was assessed by biomass inhibition studies, which confirmed that the metabolic machinery of yeast species for diesel oil degradation was plasmid coded. This is the first report establishing the involvement of a plasmid in diesel oil degradation by yeast species.

Petroleum based products are the major source of energy for industries and daily life. Leaks and accidental spills occur regularly during the exploration, production, refining, transport, and storage of petroleum and petroleum products. In the present study we isolated the bacteria from diesel contaminated soil and screened them for diesel biodegradation capacity. One monoculture isolate identified by 16S rRNA gene sequence analysis to be Acinetobacter baumannii was further studied for diesel oil biodegradation. The effects of various culture parameters (pH, temperature, NaCl concentrations, initial hydrocarbon concentration, initial inoculum size, role of chemical surfactant, and role of carbon and nitrogen sources) on biodegradation of diesel oil were evaluated. Optimal diesel oil biodegradation by A. baumanii occurred at initial pH 7, 35°C and initial hydrocarbon concentration at 4%. The biodegradation products under optimal cultural conditions were analyzed by GC-MS. The present study suggests that A. baumannii can be used for effective degradation of diesel oil from industrial effluents contaminated with diesel oil.

The performance of diesel oil degradation by Candida tropicalis immobilized on various conventional matrices (sodium alginate, carboxyl methyl cellulose, chitosan) and biowaste materials (wheat bran, sawdust, peanut hull powder) was investigated using the method of entrapment and physical adsorption. The yeast species immobilized in wheat bran showed enhanced efficiency in degrading diesel oil (98%) compared to free cells culture (80%) over a period of 7 days. Copious amount of exopolysaccharides were also produced in the presence of diesel oil. The biofilm forming ability of C. tropicalis on PVC strips was evaluated using XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide) reduction assay and monitored by scanning electron microscopy and atomic force microscopy. Yeast biofilm formed on gravels showed 97% degradation of diesel oil over a period of 10 days. The potential use of the biofilms for preparing trickling filters (gravel particles), for attenuating hydrocarbons in oily liquid wastes before their disposal in the open environment is suggested and discussed. This is the first successful attempt for 'artificially' establishing hydrocarbon degrading yeast biofilm on solid substrates.

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Efficiency of Enterobacter cloacae KU923381 isolated from petroleum hydrocarbon contaminated soil was evaluated in batch culture and bioreactor mode. The isolate were screened for biofilm formation using qualitative and quantitative assays. Response surface methodology (RSM) was used to study the effect of pH, temperature, glucose concentration, and sodium chloride on diesel degradation. The predicted values for diesel oil degradation efficiency by the statistical designs are in a close agreement with experimental data (R = 99.66%). Degradation efficiency is increased by 36.78% at pH = 7, temperature = 35°C, glucose = 5%, and sodium chloride concentration = 5%. Under the optimized conditions, the experiments were performed for diesel oil degradation by gas chromatographic mass spectrometric analysis (GC-MS). GC-MS analysis confirmed that E. cloacae had highly degrade hexadecane, heptadecane, tridecane, and docosane by 99.71%, 99.23%, 99.66%, and 98.34% respectively. This study shows that rapid bioremoval of hydrocarbons in diesel oil is acheived by E. cloacae with abet of biofilm formation. The potential use of the biofilms for preparing trickling filters (gravel particles) for the degradation of hydrocarbons from petroleum wastes before their disposal in the open environment is highly suggested. This is the first successful attempt for artificially establishing petroleum hydrocarbon degrading bacterial biofilm on solid substrates in bioreactor.

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