Microbial diversity and bioremediation of a hydrocarbon-contaminated aquifer (Vega Baja, Puerto Rico).
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Hydrocarbon contamination of groundwater resources has become a major environmental and human health concern in many parts of the world. Our objectives were to employ both culture and culture-independent techniques to characterize the dynamics of microbial community structure within a fluidized bed reactor used to bioremediate a diesel-contaminated groundwater in a tropical environment. Under normal operating conditions, 97 to 99% of total hydrocarbons were removed with only 14 min hydraulic retention time. Over 25 different cultures were isolated from the treatment unit (96% which utilized diesel constituents as sole carbon source). Approximately 20% of the isolates were also capable of complete denitrification to nitrogen gas. Sequence analysis of 16S rDNA demonstrated ample diversity with most belonging to the infinity, beta and gamma subdivision of the Proteobacteria, Bacilli, and Actinobacteria groups. Moreover, the genetic constitution of the microbial community was examined at multiple time points with a Functional Gene Array (FGA) containing over 12,000 probes for genes involved in organic degradation and major biogeochemical cycles. Total community DNA was extracted and amplified using an isothermal phi29 polymerase-based technique, labeled with Cy5 dye, and hybridized to the arrays in 50% formimide overnight at 50 degrees C. Cluster analysis revealed comparable profiles over the course of treatment suggesting the early selection of a very stable microbial community. A total of 270 genes for organic contaminant degradation (including naphthalene, toluene [aerobic and anaerobic], octane, biphenyl, pyrene, xylene, phenanthrene, and benzene); and 333 genes involved in metabolic activities (nitrite and nitrous oxide reductases [nirS, nirK, and nosZ], dissimilatory sulfite reductases [dsrAB], potential metal reducing C-type cytochromes, and methane monooxygenase [pmoA]) were repeatedly detected. Genes for degradation of MTBE, nitroaromatics and chlorinated compounds were also present, indicating a broad catabolic potential of the treatment unit. FGA's demonstrated the early establishment of a diverse community with concurrent aerobic and anaerobic processes contributing to the bioremediation process. (+info)
In situ biosurfactant production by Bacillus strains injected into a limestone petroleum reservoir.
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Biosurfactant-mediated oil recovery may be an economic approach for recovery of significant amounts of oil entrapped in reservoirs, but evidence that biosurfactants can be produced in situ at concentrations needed to mobilize oil is lacking. We tested whether two Bacillus strains that produce lipopeptide biosurfactants can metabolize and produce their biosurfactants in an oil reservoir. Five wells that produce from the same Viola limestone formation were used. Two wells received an inoculum (a mixture of Bacillus strain RS-1 and Bacillus subtilis subsp. spizizenii NRRL B-23049) and nutrients (glucose, sodium nitrate, and trace metals), two wells received just nutrients, and one well received only formation water. Results showed in situ metabolism and biosurfactant production. The average concentration of lipopeptide biosurfactant in the produced fluids of the inoculated wells was about 90 mg/liter. This concentration is approximately nine times the minimum concentration required to mobilize entrapped oil from sandstone cores. Carbon dioxide, acetate, lactate, ethanol, and 2,3-butanediol were detected in the produced fluids of the inoculated wells. Only CO(2) and ethanol were detected in the produced fluids of the nutrient-only-treated wells. Microbiological and molecular data showed that the microorganisms injected into the formation were retrieved in the produced fluids of the inoculated wells. We provide essential data for modeling microbial oil recovery processes in situ, including growth rates (0.06 +/- 0.01 h(-1)), carbon balances (107% +/- 34%), biosurfactant production rates (0.02 +/- 0.001 h(-1)), and biosurfactant yields (0.015 +/- 0.001 mol biosurfactant/mol glucose). The data demonstrate the technical feasibility of microbial processes for oil recovery. (+info)
Oil field souring control by nitrate-reducing Sulfurospirillum spp. that outcompete sulfate-reducing bacteria for organic electron donors.
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Nitrate injection into oil reservoirs can prevent and remediate souring, the production of hydrogen sulfide by sulfate-reducing bacteria (SRB). Nitrate stimulates nitrate-reducing, sulfide-oxidizing bacteria (NR-SOB) and heterotrophic nitrate-reducing bacteria (hNRB) that compete with SRB for degradable oil organics. Up-flow, packed-bed bioreactors inoculated with water produced from an oil field and injected with lactate, sulfate, and nitrate served as sources for isolating several NRB, including Sulfurospirillum and Thauera spp. The former coupled reduction of nitrate to nitrite and ammonia with oxidation of either lactate (hNRB activity) or sulfide (NR-SOB activity). Souring control in a bioreactor receiving 12.5 mM lactate and 6, 2, 0.75, or 0.013 mM sulfate always required injection of 10 mM nitrate, irrespective of the sulfate concentration. Community analysis revealed that at all but the lowest sulfate concentration (0.013 mM), significant SRB were present. At 0.013 mM sulfate, direct hNRB-mediated oxidation of lactate by nitrate appeared to be the dominant mechanism. The absence of significant SRB indicated that sulfur cycling does not occur at such low sulfate concentrations. The metabolically versatile Sulfurospirillum spp. were dominant when nitrate was present in the bioreactor. Analysis of cocultures of Desulfovibrio sp. strain Lac3, Lac6, or Lac15 and Sulfurospirillum sp. strain KW indicated its hNRB activity and ability to produce inhibitory concentrations of nitrite to be key factors for it to successfully outcompete oil field SRB. (+info)
Effects of constructed wetland system on the removal of dibutyl phthalate (DBP).
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Phthalic acid esters (PAEs) have become widely diffused in the environment via the manufacturing process. Numerous experiments have shown that the bioaccumulation of PAEs occurred in the aquatic and terrestrial food chain; meanwhile, it was found that some of PAEs were considered as potential carcinogens, teratogens and mutagens. In this research, two vertical/reverse-vertical flow constructed wetland systems were set up to study its removal efficiency of dibutyl phthalate (DBP) pollution. The results showed that the constructed wetland system could remove DBP effectively, and the removal rates reached nearly 100%. Substrate microorganism and enzymatic activities probably played key roles during DBP removal, and the removal of DBP probably mainly took place in the upper layer of chamber A in the constructed wetland systems. (+info)
Biological removal of carcinogenic chromium(VI) using mixed Pseudomonas strains.
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The contamination of soil and wastewaters with Cr(VI) is a major problem. It has been suggested that microbial methods for Cr(VI) reduction are better than chemical methods, as they do not add other ions or toxic chemicals to the environment. In this study an aerobic reduction of Cr(VI) to Cr(III) by employing mixed Pseudomonas cultures isolated from a marshy land has been reported. The role of chromium concentration, temperature, pH and additives on the microbial reduction of Cr(VI) has been investigated. NADH was found to enhance the rate of reduction of Cr(VI). Complete reduction of chromium(VI) has been possible even at chromium(VI) concentrations of 300 ppm. Ions like SO(4)(2-) and poly-phenols inhibited the metabolic activity relating to Cr(VI) reduction. Under optimal conditions 100 mg/L of Cr(VI) was completely reduced within 180 min. (+info)
Superfund: evaluating the impact of executive order 12898.
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BACKGROUND: The U.S. Environmental Protection Agency (EPA) addresses uncontrolled and abandoned hazardous waste sites throughout the country. Sites that are perceived to be a significant threat to both surrounding populations and the environment can be placed on the U.S. EPA Superfund list and qualify for federal cleanup funds. The equitability of the Superfund program has been questioned; the representation of minority and low-income populations in this cleanup program is lower than would be expected. Thus, minorities and low-income populations may not be benefiting proportionately from this environmental cleanup program. In 1994 President Clinton signed Executive Order 12898 requiring that the U.S. EPA and other federal agencies implement environmental justice policies. These policies were to specifically address the disproportionate environmental effects of federal programs and policies on minority and low-income populations. OBJECTIVE AND METHODS: I use event history analysis to evaluate the impact of Executive Order 12898 on the equitability of the Superfund program. DISCUSSION: Findings suggest that despite environmental justice legislation, Superfund site listings in minority and poor areas are even less likely for sites discovered since the 1994 Executive Order. CONCLUSION: The results of this study indicate that Executive Order 12898 for environmental justice has not increased the equitability of the Superfund program. (+info)
Protecting contract workers: case study of the US Department of Energy's nuclear and chemical waste management.
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Increased reliance on subcontractors in all economic sectors is a serious occupational health and safety challenge. Short-term cost savings are offset by long-term liability. Hiring subcontractors brings specialized knowledge but also young, inexperienced, inadequately trained workers onto industrial and hazardous waste sites, which leads to increased rates of accidents and injuries. Reliable data on subcontractor occupational health and safety programs and performance are sparse. The US Department of Energy has an excellent safety culture on paper, but procurement practices and contract language deliver a mixed message--including some safety disincentives. Its biphasic safety outcome data are consistent with underreporting by some subcontractors and underachievement by others. These observations are relevant to the private and public sectors. Occupational health and safety should be viewed as an asset, not merely a cost. (+info)
The effect on ecological systems of remediation to protect human health.
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Environmental remediation of contaminated eco-systems reduces stresses to these ecosystems, including stresses caused by the production, use, and storage of weapons of mass destruction. The effects of these various stressors on humans can be reduced by remediation or by blocking the exposure of humans, but blocking the exposure of resident biota is almost impossible. Remediation may involve trade-offs between reducing a minor risk to public health and increasing risks to workers and ecosystems. Remediation practices such as soil removal disrupt ecosystems, which take decades to recover. Without further human disturbances, and with low levels of exposure to stress-ors, ecosystems can recover from physical disruptions and spills. Remediation to remove negligible risk to humans can destroy delicate ecosystems for very little gain in public health. (+info)