Improvement of Pure Poly aromatic Hydrocarbon Degrading Bacteria Using Gamma Irradiation

The main goal of this study is to obtain a potent polycyclic aromatic hydrocarbons (PAHs) biodegrading bacteria to be used for bio augmentation purpose which is considered a promising strategy for the cleanup of contaminated sites. Among 10 selected potent PAHs degrading bacteria isolated from Suez Gulf water (after enrichments on phenanthrene as a sole source of carbon and energy). Isolate HD20 was selected due to its fast and remarkable abilities to breakdown phenanthrene . In a trial to improve the biodegradation potentials of the most potent isolate (Had) identified as Stenotrophomonas maltophilia, low doses of gamma irradiation were used to activate the organism. To determine the radiation response of S. maltophilia, 24 hours culture was exposed in saline solution in a triplicate glass vials separately to increasing doses of gamma irradiation, 0.25, 0.5, 1.0, 1.5, 2.0 and 2.5 kGy. The dose response curve revealed the linear death of bacterial cells with increasing irradiation dose, The D10 value of S. maltophilia was found to be 0.3 kGy. For PAHs biodegradation enhancement, the organism was irradiated at low doses of gamma irradiation, 0.2, 0.4, 0.5, 0.6 and 0.7 kGy. The grown colonies exhibited a morphological differences from the non irradiated Stenotrophomonas maltophilia on Tryptone-glucose-yeast extract (TGY) plates. All of them showed a creamy rough appearance and a loss of the known yellow colour of the original isolate. The single selected irradiated colonies as well as the whole irradiated mixture of cells irradiated at different doses were tested separately in fertilized marine microcosms containing 200 ppm of phenanthrene as a model of PAH and the degradation rates of polyaromatic hydrocarbons were monitored by the determination of the residual phenanthrene up to one week . The biodegradation potentials of irradiated and non irradiated S. maltophilia was compared with that of the natural Suez Gulf microbial communities. The irradiated culture of S. maltophilia at dose 0.7 kGy (M7) showed the maximum degradation potential (31.5%) after 96 hrs compared to 18.5% after 168 hours degradation for the control (non irradiated isolate). whereas the selected single isolate at dose of 0.7 kGy (S7) shows its maximum biodegradation potential (30%) after 72 hrs. The biodegradation potentials of the non irradiated adapted natural mixed microflora of Suez Gulf was found to be 19%. According to the findings of this study, it is recommended to use both the 0.7 kGy-irradiated cell mixture of S. maltophilia as well as the single colonies of the organism at the same irradiation dose to examine their bio augmentation efficiency.