Isolation and Molecular study of indigenous Bacillus strains from oil storage | Abstract
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Isolation and Molecular study of indigenous Bacillus strains from oil storage

Author(s): Sevil Hariri Ahari*1, Behboud Jafari1, Morteza Hanifehzadeh2

Iran is first country in oil rich in the Middle East region that operation of its oil productioncapacity more than 4 barrels of crude oil and m3/day80000 diesel fuel began. In this study, isolation and characterization of bacterial strains isolated from soil contaminated with hydrocarbons investigated in the oil distribution centers and stores in Ahar throughout the years 2010 to2011. The main purpose of this article is describe the bacteria isolation that have ability analyze and gene involved in the analyze of diesel. Bacterial strains used in this study of soils contaminated with crude oil hydrocarbons in the oil distribution centers and stores in Ahar-Iran were isolated. 5 samples soil infected to diesel oil were collected from underground storage area and were kept at 4c in closed containers for use. Commercial diesel fuel as the only carbon source was added to the autoclaved medium. PH solid medium for each of Hcl or Nacl is set to 7/3. The enriched medium was used to isolate the diesel-degrading bacteria. Basis mineral medium(MM) containing of soil and diesel was incubated at 35°C with orbital vibration (120r/min). All strains to determine the ability analyzed diesel fuel and growth were tested in rich liquid medium. A collection of Erlen Mayerflaks 500ml was used for this experiment. Each flak containing 100ml of 8g/l nutrient broth plus diesel and medium with transfer 3ml of pre-culture were incubated and primary PH was adjusted to 7/3. Genomic DNA isolationwas extracted with an extraction kit from sigma.Filogenic related isolates, were determined by comparing sequence date with sequences of members of the genus Bacillus that is available through the database,Genbank, National center for Biotechnology information. Each gene sequences from isolates obtained in this study compared with GenBank database sequences in the gene 16srRNA. Biochemical and physiological tests performed is species test, Bacillus sp, TMY-z. To confirm the identity of isolation, PCR and sequencing was performed in 16 srRNA. To verify the identity of bacteria identified as Bacillus niacin with phenotypic analysis. Benzoevorans, oleronius, Galactosidilyticus strains is a potential volunteer for bioremediation of sites contaminated by the diesel fuel. This species identified parser effectively as Bacillus niacin (more than 98% similarity). Using consortium of indigenous bacteria with the ability to use diesel as seed into the environment infected with oil could prove environmentally conducive approach that cause increases sustainable development bacterial species are foreign and chemicals.