Dataset Information

Molecular characterization of bacterial populations in petroleum-contaminated groundwater discharged from underground crude oil storage cavities.

ABSTRACT: Petroleum-contaminated groundwater discharged from underground crude oil storage cavities (cavity groundwater) harbored more than 10(6) microorganisms ml(-1), a density 100 times higher than the densities in groundwater around the cavities (control groundwater). To characterize bacterial populations growing in the cavity groundwater, 46 PCR-amplified almost full-length 16S ribosomal DNA (rDNA) fragments were cloned and sequenced, and 28 different sequences were obtained. All of the sequences were affiliated with the Proteobacteria; 25 sequences (43 clones) were affiliated with the epsilon subclass, 2 were affiliated with the beta subclass, and 1 was affiliated with the delta subclass. Two major clusters (designated clusters 1 and 2) were found for the epsilon subclass proteobacterial clones; cluster 1 (25 clones) was most closely related to Thiomicrospira denitrificans (88% identical in nucleotide sequence), while cluster 2 (11 clones) was closely related to Arcobacter spp. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified partial 16S rDNA fragments showed that one band was detected most strongly in cavity groundwater profiles independent of storage oil type and season. The sequence of this major band was identical to the sequences of most of the cluster 1 clones. Fluorescence in situ hybridization (FISH) indicated that the cluster 1 population accounted for 12 to 24% of the total bacterial population. This phylotype was not detected in the control groundwater by DGGE and FISH analyses. These results indicate that the novel members of the epsilon subclass of the Proteobacteria grow as major populations in the petroleum-contaminated cavity groundwater.

SUBMITTER: Watanabe K

PROVIDER: S-EPMC92383 | biostudies-literature | 2000 Nov

REPOSITORIES: biostudies-literature

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Molecular characterization of bacterial populations in petroleum-contaminated groundwater discharged from underground crude oil storage cavities.

Watanabe K K Watanabe K K Kodama Y Y Syutsubo K K Harayama S S

Applied and environmental microbiology 20001101 11

Petroleum-contaminated groundwater discharged from underground crude oil storage cavities (cavity groundwater) harbored more than 10(6) microorganisms ml(-1), a density 100 times higher than the densities in groundwater around the cavities (control groundwater). To characterize bacterial populations growing in the cavity groundwater, 46 PCR-amplified almost full-length 16S ribosomal DNA (rDNA) fragments were cloned and sequenced, and 28 different sequences were obtained. All of the sequences wer ...[more]

PMID: 11055927

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Project description:This study investigated the impacts of crude oil, diesel, and gasoline on the diversity of indigenous microbial communities as well as culturable microorganisms in the studied soil. Oil contamination led to shifts in the diversity of the soil's microbial communities, regardless of the contaminant applied. Unpolluted soils were more diverse and evenly distributed than contaminated samples. The domain Bacteria accounted for 65.15% of the whole microbial community. The bacterial phylum Proteobacteria dominated in all samples, followed by Actinobacteria and Acidobacteria. Pseudomonas with 28.15% of reads dominated in Proteobacteria, while Rhodococcus (3.07%) dominated in Actinobacteria, and Blastocatella (2.53%) dominated in Acidobacteria. The dominant fungal phyla across all samples were Ascomycota dominated by Penicillium (50.48% of sequences), and Zygomycota dominated by Mortierella (16.87%). Sequences similar to the archaeal phyla, Euryarchaeota and Thaumarchaeota, were also detected. The number of culturable microorganisms increased following the contamination and was higher in contaminated samples than in clean samples. Oil contamination also resulted in the enrichment of oil-degrading strains. Two bacteria, Serratia marcescens strain PL and Raoultella ornithinolytica PS, which were isolated from crude oil-contaminated soil, exhibited strong crude oil degradation ability. Strain PL was the most efficient strain and degraded 75.10% of crude oil, while strain PL degraded 65.48%, after 20 days of incubation. However, the mixed culture of the two strains was more effective than single strain and could achieve up to 96.83% of crude oil degradation, with a complete abatement of straight-chain hydrocarbons (from C12 to C25), and more than 91% removal of highly branched hydrocarbons, phytane and pristane, which are known to be more recalcitrant to biodegradation. Strains PS and PL are two newly isolated crude oil degraders that are not among the most prominent crude oil-degrading strains referenced in the literature. Therefore, their high degradation capacity makes them perfect candidates for the bioremediation of petroleum hydrocarbon contaminated environments.

Project description:Total petroleum hydrocarbons (TPHs), which are often found in soil, water, sediments, and air. These compounds are a type of pollutant that can have a serious negative impact on living things and human health. Soil washing method is a remediation technique used to remove contaminants from the soil. This process involves the use of water or other solvents to extract contaminants from the soil, followed by separation and disposal of the contaminated solution. This research engineered the effectiveness of soil washing method to remove TPHs from a genuine, sullied soil sample. After analyzing the physical and chemical properties of the soil, the Box-Benken Design (BBD) technique was used to optimize the variables that influence the process's effectiveness. A quadratic model was suggested based on the BBD design, correlation coefficients, and other factors. The minimum, maximum and mean removal of TPHs during the stages of the study were 63.5, 94.5 and 76.7%, respectively. The correlation between the variables was strong, as shown by the analysis of variance (ANOVA), F-value (1064.5) and P-value (0.0001), and the proposed model was highly significant. The most effective soil washing method (SWM) was obtained with pH 7.8, liquid to solid ratio 50:1, reaction time 52 min, surfactant concentration 7.9 mg kg-1, and three washings. A removal rate of 98.8% was accomplished for TPHs from the soil in this context. The kinetic results indicate that the kinetic of TPHs removal follows the first-order kinetics (R2 = 0.96). There was not a major difference in the process's efficiency based on temperature. The removal efficiency heightened from 0 to 150 rpm and then remained steady. Introducing air flow increased the rate of removal, and the combination of ultrasonic waves with the reaction environment increased the process efficiency and decreased the time for the process and the amount of times it needed to be washed. An analysis of the washed soil both physically and chemically revealed a substantial decrease in the concentration of other elements.

Dataset Information

Molecular characterization of bacterial populations in petroleum-contaminated groundwater discharged from underground crude oil storage cavities.

Publications

Molecular characterization of bacterial populations in petroleum-contaminated groundwater discharged from underground crude oil storage cavities.

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