Molecular Characterization of Petroleum Compounds-Degrading Bacteria Isolated from Petroleum Contaminated Soil
Mohamed Eraky Mohamed Wahballah;
Abstract
Oil contamination with petroleum and petroleum-based hydrocarbons
has caused critical environmental and health defects and increasing attention has been paid for developing and implementing innovative technology for cleaning up this contaminant. Bioremediation methods are currently receiving favorable publicity as promising environmental friendly treatment technologies for the remediation of hydrocarbons. Moreover, biological methods can have an edge over the physico-chemical treatment regimes in removing spills as they offer cost effective in situ biodegradation of oil fractions by the microorganisms. Bioremediation can be described as the conversion of chemical compounds by living organisms, especially microorganisms, into energy, cell mass and biological waste products. The results of this study were summeraized as follow:
1- The bacterial strains used in this study were isolated from contaminated soil near gas stations.
2- The bacterial strains were isolated by enrichment cultivation from oil-contaminated soil samples on Modified Basal Salt Medium (MBSM) amended with 2% crude oil, (50 mg/L) Benzene, toluene, ethylbenzene, p-xylene, o-xylene, m-xylene, respectively and BTEX mixture (1:1:1:1:1:1) as the sole source of carbon and energy.
3- According to the colony size, twenty bacterial isolates were obtained by their enrichment on 2% crude oil while thirty four bacterial isolates were obtained by enrichment on BTEX compounds.
SUMMARY
101
4- The bacterial isolates able to grow on 50 mg/L BTEX or 2% crude oil were screened for their growth on different concentrations of BTEX and 4% crude oil respectively.
5- The isolate MS30 was selected based on its ability to grow on MBSM plates amended with 250 mg/L benzene, toluene, ethylbenzene, o-, m- and p-xylene individually.
6- Four bacterial isolates (RAM03, RAM06, RAM13 and RAM17) were selected based on their growth rates on 4% crude oil.
7- The analysis of 16SrRNA gene sequences confirmed that the isolate RAM03 revealed its affiliation to Ochrobactrum cytisi, RAM06 and RAM17 revealed their affiliation to Ochrobactrum anthropi, RAM13 and MS30 revealed their affiliation to Sinorhizobium meliloti and Ochrobactrum lupine with 96, 97, 97, 97 and 97% similarity, respectively.
8- From the growth curves of the selected bacterial isolates on MBSM containing 2% crude oil, O. cytisi (RAM03) was achieved the highest log10 cfu/ml after 3 days incubation at 30◦C.
9- O. cytisi (RAM03) and O. anthropi (RAM06) had good growth on 200 mg/L of Benzene, Toluene, Ethylbenzene, m- and p- Xylenes and BTEX mixture but their was no significant growth on o-Xylene.
10- The highest growth rates of the bacterial strains O. cytisi (RAM03), O. anthropi (RAM06) and O. anthropi (RAM17) was obtained at pH 7, while Sinorhizobium meliloti (RAM13) and O. lupine (MS30) prefered pH 8 in MBSM amended with 100mg/L BTEX at 30◦C.
SUMMARY
102
11- According to growth rate of the bacteria on MBSM amended with 100 mg/L BTEX, The most suitable temperature of the bacterial growth was 30◦C.
12- The bacterial strains used in this study were able to produce biosurfactant causing high emulsification index and decreasing of the surface tension and enhance availability and the utilization of the
petroleum compounds by the microorganisms.
13- Ochrobactrum sp. (RAM03, RAM06, RAM17 and MS30) and Sinorhizobium meliloti (RAM13) degraded more than 84% of the TPHs and 80-100% of n-alkanes (C14-C30) after 30 day incubation period in MBSM supplemented with 4% crude oil at ◦C.
14- Ochrobactrum sp (RAM03, RAM06, RAM13 and MS30) and Sinorhizobium meliloti (RAM17) were able to completely degrade chrysene (100%) and degrade > 80% of phenantherene and > 48% of (anthracene) found in MBSM amended with 4% crude oil after 30 day at 30◦C.
15- O. cytisi (RAM03) degraded 54.3% of TPH found in the soil while the other bacterial isolates degraded 19-49.2% in batch experiment.
16- O. cytisi (RAM03) degraded almost all n-alkanes (C14-C30) present in the crude oil. In contrast, O. anthropi (RAM06) had low n-alkane degradation abilities (in the batch study).
17- C14 was increased in the batch study after incubation of O. anthropi (RAM06) for 30 days.
18- Benzo(b)fluoranthene was degraded completely after 30day by O. cytisi (RAM03), Sinorhizobium meliloti (RAM13), O. anthropi (RAM17) and
SUMMARY
103
O. lupine (MS30) while O. anthropi (RAM06) degraded 84.25% of Benzo(b)fluoranthene. Pyrene was degraded (52.9, 74.08, 81.5, 85.5 and 48.5%) by RAM03, RAM06, RAM13, RAM17 and MS30 respectively (in the batch study).
These bacterial strains obtained could effectively remove both aliphatic and aromatic petroleum hydrocarbons, and they are able to produce bio-surfactant. However, these data indicate that these isolates may have the potential for use in bioremediation of petroleum hydrocarbon contaminated soil.
has caused critical environmental and health defects and increasing attention has been paid for developing and implementing innovative technology for cleaning up this contaminant. Bioremediation methods are currently receiving favorable publicity as promising environmental friendly treatment technologies for the remediation of hydrocarbons. Moreover, biological methods can have an edge over the physico-chemical treatment regimes in removing spills as they offer cost effective in situ biodegradation of oil fractions by the microorganisms. Bioremediation can be described as the conversion of chemical compounds by living organisms, especially microorganisms, into energy, cell mass and biological waste products. The results of this study were summeraized as follow:
1- The bacterial strains used in this study were isolated from contaminated soil near gas stations.
2- The bacterial strains were isolated by enrichment cultivation from oil-contaminated soil samples on Modified Basal Salt Medium (MBSM) amended with 2% crude oil, (50 mg/L) Benzene, toluene, ethylbenzene, p-xylene, o-xylene, m-xylene, respectively and BTEX mixture (1:1:1:1:1:1) as the sole source of carbon and energy.
3- According to the colony size, twenty bacterial isolates were obtained by their enrichment on 2% crude oil while thirty four bacterial isolates were obtained by enrichment on BTEX compounds.
SUMMARY
101
4- The bacterial isolates able to grow on 50 mg/L BTEX or 2% crude oil were screened for their growth on different concentrations of BTEX and 4% crude oil respectively.
5- The isolate MS30 was selected based on its ability to grow on MBSM plates amended with 250 mg/L benzene, toluene, ethylbenzene, o-, m- and p-xylene individually.
6- Four bacterial isolates (RAM03, RAM06, RAM13 and RAM17) were selected based on their growth rates on 4% crude oil.
7- The analysis of 16SrRNA gene sequences confirmed that the isolate RAM03 revealed its affiliation to Ochrobactrum cytisi, RAM06 and RAM17 revealed their affiliation to Ochrobactrum anthropi, RAM13 and MS30 revealed their affiliation to Sinorhizobium meliloti and Ochrobactrum lupine with 96, 97, 97, 97 and 97% similarity, respectively.
8- From the growth curves of the selected bacterial isolates on MBSM containing 2% crude oil, O. cytisi (RAM03) was achieved the highest log10 cfu/ml after 3 days incubation at 30◦C.
9- O. cytisi (RAM03) and O. anthropi (RAM06) had good growth on 200 mg/L of Benzene, Toluene, Ethylbenzene, m- and p- Xylenes and BTEX mixture but their was no significant growth on o-Xylene.
10- The highest growth rates of the bacterial strains O. cytisi (RAM03), O. anthropi (RAM06) and O. anthropi (RAM17) was obtained at pH 7, while Sinorhizobium meliloti (RAM13) and O. lupine (MS30) prefered pH 8 in MBSM amended with 100mg/L BTEX at 30◦C.
SUMMARY
102
11- According to growth rate of the bacteria on MBSM amended with 100 mg/L BTEX, The most suitable temperature of the bacterial growth was 30◦C.
12- The bacterial strains used in this study were able to produce biosurfactant causing high emulsification index and decreasing of the surface tension and enhance availability and the utilization of the
petroleum compounds by the microorganisms.
13- Ochrobactrum sp. (RAM03, RAM06, RAM17 and MS30) and Sinorhizobium meliloti (RAM13) degraded more than 84% of the TPHs and 80-100% of n-alkanes (C14-C30) after 30 day incubation period in MBSM supplemented with 4% crude oil at ◦C.
14- Ochrobactrum sp (RAM03, RAM06, RAM13 and MS30) and Sinorhizobium meliloti (RAM17) were able to completely degrade chrysene (100%) and degrade > 80% of phenantherene and > 48% of (anthracene) found in MBSM amended with 4% crude oil after 30 day at 30◦C.
15- O. cytisi (RAM03) degraded 54.3% of TPH found in the soil while the other bacterial isolates degraded 19-49.2% in batch experiment.
16- O. cytisi (RAM03) degraded almost all n-alkanes (C14-C30) present in the crude oil. In contrast, O. anthropi (RAM06) had low n-alkane degradation abilities (in the batch study).
17- C14 was increased in the batch study after incubation of O. anthropi (RAM06) for 30 days.
18- Benzo(b)fluoranthene was degraded completely after 30day by O. cytisi (RAM03), Sinorhizobium meliloti (RAM13), O. anthropi (RAM17) and
SUMMARY
103
O. lupine (MS30) while O. anthropi (RAM06) degraded 84.25% of Benzo(b)fluoranthene. Pyrene was degraded (52.9, 74.08, 81.5, 85.5 and 48.5%) by RAM03, RAM06, RAM13, RAM17 and MS30 respectively (in the batch study).
These bacterial strains obtained could effectively remove both aliphatic and aromatic petroleum hydrocarbons, and they are able to produce bio-surfactant. However, these data indicate that these isolates may have the potential for use in bioremediation of petroleum hydrocarbon contaminated soil.
Other data
| Title | Molecular Characterization of Petroleum Compounds-Degrading Bacteria Isolated from Petroleum Contaminated Soil | Other Titles | التوصيف الجزيئي للبكتريا المحللة للمواد البترولية والمعزولة من تربة ملوثه | Authors | Mohamed Eraky Mohamed Wahballah | Issue Date | 2015 |
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