Biodegradation of Crude Oil Spill Using Bacillus Subtilis and Pseudomonas Putida in Sequencing Method

Crude oil, otherwise called petroleum, occurs naturally as a complex organic mixture underneath the subsurface. The activities related to its exploration, production, refining, storage and distribution are mostly accompanied with extreme pollution and other hazardous conditions. For these reasons, the need to critically devise the best possible solutions becomes paramount, particularly as regards oil spills. Therefore, the purpose of this research was to determine the efficiency of TPH removal in crude oil using Bacillus Subtilis and Pseudomonas Putida. The sequencing method was applied in a laboratory scale and under artificial seawater media conditions. The total petroleum hydrocarbon (TPH) serves as a significant parameter in detecting crude oil, although the extraction and analysis were conducted with the use of a separator funnel and gas chromatography mass spectrometry (GCMS), respectively. In addition, the simulated seawater media was described as the mineral salt medium (MSM), with 33% salinity. Moreover, five reactors were also employed, including K for control, B for B. subtilis, P for P. putida, BP for B. subtilis and P. putida sequence and PB for P. putida and B. subtilis sequence. The entire treatments obtained the access to two replicate reactors. Furthermore, the bacteria inoculum and crude oil concentration in each unit were estimated at 5% and 10% (v/v), respectively. The results achieved the maximum TPH removal at 66.29% in the PB reactor after 35 days. On the basis of ANOVA reports, no significant variation was observed between the sequential additions of a single bacterial treatment and consortium microbes. In summary, two bacterial species demonstrated high potential to degrade TPH, but predicted an increase in the break down time, as the nutrient or oxygen tends to accelerate the process.