|Budget Amount *help
¥4,100,000 (Direct Cost : ¥4,100,000)
Fiscal Year 2003 : ¥1,600,000 (Direct Cost : ¥1,600,000)
Fiscal Year 2002 : ¥2,500,000 (Direct Cost : ¥2,500,000)
Bioremediation has been partially employed as a technical option to remove spilled oil at some coastal and soil environments. In almost of all cases, biostimulation in which indigenous pollutants-decomposing microorganisms are enriched and/or activated by the exogenous supply of nutrients has been applied instead of bioaugmentation. The essence of spilled oil bioremediation is exogenous supply of macronutrients to indigenous hydrocarbon-degrading microorganisms. However, the efficacy to exogenously supply of macronutrients remains to be controversial. Recently some methods like DGGE and clone library analysis of 16S rDNA are widely used to chase the behaviors and occurrences of the key hydrocarbon-degrading bacteria in evaluating oil bioremediation processes. Unfortunately, these tools give us qualitative information, not quantitative one. Therefore, in this research, we constructed PCR primer sets to detect oil-degrading bacteria specifically, and traced the transition of oil-degrading bacteria in biostimulation process by using real-time PCR.
Consequently, we succeeded to construct the PCR primer sets to detect representative genes in oil-degrading pathway, alkB, C120, and C230, encoding alkane hydroxylase, catechol 1,2 dioxygenase, and catechol 2,3 dioxygenase, respectively. In the field experiments, it was shown that bacteria possessing alkB and C120 were dominant in oil degradation process. These 3 primer sets were also effective for soil samples. These results suggest that our primer sets are feasible for the use in evaluating effectiveness and safety of bioremediation.