Information Last Revised: 2-15-93
TTP Reference Number: OR 1111-01 (3BAC)
1. Technical Name of Technology: Microbial Monitoring
2. Common Name of Technology: Microbial Monitoring
3. PI and Telephone No: Anthony B. Palumbo 615-576-8002
4. Affiliation: Oak Ridge National Laboratory
5. Technology Category: Monitoring and Characterization
6. Developers: VOC's at Non-Arid Sites/ ORNL/ University of Tennessee/ University of Minnesota
7. Application
7.1. Where (in-situ/ex-situ): in-situ and ex-situ
7.2. Media: Soil, Groundwater, Bioreactors
7.3. Targeted Contaminants: TCE, PCE, Toluene
8. Scope of project (feasibility study, treatability, bench, pilot, field):
Application to all phases
9. Integrated Demonstration (ID) Need/Requirements:
DOE sites have been contaminated with numerous organic compounds. Many technologies designed to remediate this contamination either directly depend on increasing populations of bacteria that can degrade the compounds (such as the methane injection at SRL) or lead to increases in degradative populations as a secondary result of non-biological remediation (such as the biological benefits of venting-bioventing). This technology is needed to demonstrate the effectiveness of bioremediation and for demonstration of additional bioremediation benefits from other technologies (e.g. bioventing benefits from soil venting). The various advanced monitoring techniques developed and applied as part of this task (DNA probe analysis, lipid analysis, activity and biomass measurements) all contribute to documenting the necessary changes in microbial populations. In addition, these techniques afford the opportunity to give feedback during operation so that procedures may be changed (e.g. changing nutrients) to increase effectiveness of the remediation.
10. Objective
10.1. Objective of technology (i.e., This technology will destroy VOCs in groundwater.):
This will contribute evidence for biological destruction of VOC's. The objective is to develop and demonstrate the techniques that can be used to monitor significant changes in the microbial populations during remediation. Significant changes are those that result in increased or decreased effectiveness of the remediation.
10.2. Baseline (baseline technology to which it is compared):
Mass Balance Measurements (Mass Balance)
11. Process Description:
This project monitors microbial population changes in soil and groundwater samples form SRS. Microbial biomass was evaluated using measurements of colony forming units, MPN techniques for methanotroph and methylotroph population and PFLA analysis. Microbial activity was assessed using acetate incorporation techniques and by measurement of TCE degradation in enrichments. DNA probes are used to enumerate specific groups of bacteria such as methanotrophs, toluene degraders, etc. Several new probes have been developed as part of this work.
The primary source of samples for the monitoring is a series of 122 wells located at and around the methane injection site at Savannah River. The wells are sampled biweekly and transferred to Tennessee for processing. Samples are then held at reduced temperature until processed. Additional samples come from periodic drilling and coreing procedures.
11.1. Input:
11.2. Output:
The output consists of data reflecting changes in microbial populations.
12. Summary of Technology Advantages (relative to the baseline: faster, better, cheaper, safer):
The limitations of mass balances approaches with field sites have led to efforts designed to monitor critical changes in the microbial populations that would result in increased rates of TCE degradation. If the remediation technique is a biological activity information on the progress and the success of the remediation activities can come from monitoring changes in critical microbial populations. An additional benefit is the feedback on a more fundamental level than is available from mass balance approaches. This feedback can result in better control of the process resulting in more effective remediation. Evaluation of the effectiveness of a large scale remediation of chlorinated solvents in the subsurface can be a significant problem given uncertainties usually present in estimating total concentrations of contaminant present. These limitations often result in a failure of the mass balance approach.
13. Limitations of Technology (relative to the baseline: faster, better, cheaper, safer):
Biological monitoring will add costs at the monitoring stage. However, it can result in reduced total costs by affording the potential for a faster and cheaper remediation effort by increasing efficiency of the remediation.
14. Major Technical Challenges:
Major challenges have included development of DNA probes appropriate for the critical populations, application of DNA concentration and extraction techniques, processing large numbers of samples required for effective monitoring and interpretation of results in a timely manner to afford feedback for the remediation efforts.
15. Technical Effectiveness:
The monitoring has been very effective in documenting significant changes in the microbial community in response to the remediation. The results of this effort indicate that there have been substantial changes in biological activity and biomass with the increasingly aggressive measures to promote TCE degration in the subsurface. However, there are some indications of a leveling off or a decrease in some of these measures as the 4% methane injection proceeds. Thus the data indicate the success in stimulating TCE degrading populations in the subsurface and now may be indicating a limitation of further increases or the potential for decreases in critical populations. Other nutrients may be becoming limiting and a further phase (methane injection with nutrient addition) may be necessary to further increase critical population levels and degradative activity.
15.1. Performance
15.1.1. Remaining Contamination: (contamination mobility reduction, volume reduction, toxicity reduction)
Summary (20 words or less): Not applicable for monitoring technology.
Further Description (unlimited length):
15.1.2. Process Waste
15.1.2.1. Status of waste (mobility, volume, hazard, recyclability)
Summary (20 words or less): Small amounts of contaminated material (soil and groundwater) are used in the monitoring. This material must be disposed of as hazardous waste.
Further Description (unlimited length):
15.1.2.2. Treatment (needed, available)
Summary (20 words or less): Disposal is via Oak Ridge National Labs guidelines.
Further Description (unlimited length):
15.1.2.3. Decontamination / Decommissioning
Summary (20 words or less): No significant decontamination or decommissioning is required.
Further Description (unlimited length):
15.1.2.4. Disposal (needed, available)
Summary (20 words or less): Disposal is standard for contaminated soil and groundwater samples.
Further Description (unlimited length):
15.1.3. Practicality
15.1.3.1. Foreclose Future Options
Summary (20 words or less): No effects on future options
Further Description (unlimited length):
15.1.3.2. Reliability
Summary (20 words or less): Instruments (e.g. gas chromatograph / mass specrometer used in lipid analysis) must be maintained on a regular basis.
Further Description (unlimited length):
15.1.3.3. Failure Control
Summary (20 words or less): No immediate consequences.
Further Description (unlimited length):
15.1.3.4. Ease of Use
Summary (20 words or less): The techniques must be applied by trained personnel.
Further Description (unlimited length):
15.1.3.5. Infrastructure
Summary (20 words or less): Depending on the techniques used, specific instruments are needed.
Further Description (unlimited length): Instruments needed could include gas chromatograph / mass specrometer, liquid scintillation counters, auto-analyzers, and a well equipped molecular biology laboratory.
15.1.3.6. Versatility
Summary (20 words or less): Techniques can be modified to monitor degradation of most organic contaminants.
Further Description (unlimited length): Probe development and other modifications may be needed for specific contaminants.
15.1.3.7. System Compatibility
Summary (20 words or less): Not applicable
Further Description (unlimited length):
15.1.3.8. Off-the-Shelf (procurement ease)
Summary (20 words or less): Equipment and materials are off the shelf except for DNA probes, which must be made or obtained from researchers.
Further Description (unlimited length):
15.1.3.9. Maintainability
Summary (20 words or less): Periodic maintenance is required for instruments used in the monitoring.
Further Description (unlimited length):
15.1.3.10. Safety Measures
Summary (20 words or less): Standard laboratory and field sampling procedures are required.
Further Description (unlimited length):
15.1.4. "Works" (functions as intended):
Summary (20 words or less): Has provided important data for the SRS Integrated Demonstration.
Further Description (unlimited length):
15.2. Cost
15.2.1. Start-Up Cost
Summary (20 words or less): Will vary with technique and pre-existing equipment.
Further Description (unlimited length): Highest start up cost would be for PFLA analysis if gas chromatograph / mass specrometer purchase is necessary. CFU, Most Probable Number, activity measurement start up costs are minimal if done in an existing microbiology laboratory. DNA probe start up costs are low provided development of new probes is not necessary.
15.2.2. Operations and Maintenance Cost
Summary (20 words or less): Depending on techniques, the per sample cost can vary from less than $100 to more than $500.
Further Description (unlimited length): Higher costs are associated with PFLA analysis. Costs will vary with the intensity of the sampling techniques used. Costs at SRS were high due to redundancy built in to sampling needed to develop the data on reliability of individual monitoring techniques. Costs for a more focused effort would be lower.
15.2.3. Life-cycle cost
Summary (20 words or less): Depends on intensity and duration of sampling.
Further Description (unlimited length):
15.3. Time
15.3.1. Years Until Available
Summary (20 words or less): Available now for contaminants used to date.
Further Description (unlimited length):
15.3.2. Speed/Rate
Summary (20 words or less): Analysis is complete in 2 days to 1 month depending on techniques used.
Further Description (unlimited length): For specific contaminants DNA probe development may be needed at 6 months to 1 year depending on contaminants.
15.3.3. Years to Finish
Summary (20 words or less): Depends on length of remediation effort monitoring and post monitoring requirements.
Further Description (unlimited length):
16. Environmental Safety and Health
16.1. Worker Safety
16.1.1. Exposure to Hazardous Materials/Hazards
Summary (20 words or less): Potential exists for exposure to solvents and low levels of radionuclides (e.g. 14C).
Further Description (unlimited length): Worker health and safety issues are associated with standard laboratory and field sampling operations.
16.1.2. Physical Requirements
Summary (20 words or less): No unusual physical requirements.
Further Description (unlimited length):
16.1.3. Number of People Required
Summary (20 words or less): Number of people required depends on the number of techniques used and intensity of sampling. Minimum is probably two people.
Further Description (unlimited length):
16.2. Public Health and Safety
16.2.1. Accidents
Summary (20 words or less): No public health and safety issues have been identified.
Further Description (unlimited length):
16.2.2. Routine Releases
Summary (20 words or less): No public health and safety issues have been identified.
Further Description (unlimited length):
16.2.3. Transportation
Summary (20 words or less): Transportation of samples to the laboratory is essential for the analysis. DOT standards are followed.
Further Description (unlimited length):
16.3. Environmental Impacts
16.3.1. Ecological Impacts
Summary (20 words or less): No ecological impacts are anticipated.
Further Description (unlimited length):
16.3.2. Aesthetics
Summary (20 words or less): No impacts are associated with this technology.
Further Description (unlimited length):
16.3.3. Natural Resources
Summary (20 words or less): No impacts except those associated with wells and drilling for obtaining samples.
Further Description (unlimited length):
16.3.4. Energy Demands
Summary (20 words or less): Energy use is minimal.
Further Description (unlimited length):
17. Socio-Political Interests
17.1. Public Perception
17.1.1. Proponent Reputation
Summary (20 words or less): Although these techniques use hazardous chemicals and radioisotopes there has been no problem in the past.
Further Description (unlimited length):
17.1.2. Familiarity / Understandability
Summary (20 words or less): This technology is common and can be explained to a non-technical audience in terms of the general principals involved.
Further Description (unlimited length):
17.2. Tribal Rights / Future Land Use
17.2.1. Capacity for Unrestricted Use (terrestrial, aquatic)
Summary (20 words or less): Wellheads may impact terrestrial use if post remediation monitoring is required. This should not be the case for most/any situations.
Further Description (unlimited length):
17.3. Socio-Economic Interests
17.3.1. Economic Impacts
Summary (20 words or less): No impacts are anticipated.
Further Description (unlimited length):
17.3.2. Labor Force Demands
Summary (20 words or less): No significant demands are anticipated.
Further Description (unlimited length):
18. Regulatory Objectives
18.1. Compatibility with Cleanup Milestones
Summary (20 words or less): Not usually directly relevant to clean-up milestones.
Further Description (unlimited length):
18.2. Regulatory Infrastructure / Track Record
Summary (20 words or less): To our knowledge no EPA standards for these techniques exists.
Further Description (unlimited length):
18.3. Regulatory Compliance
Summary (20 words or less): National Environmental Policy Act (NEPA) documentation is in place for these activities.
Further Description (unlimited length):
19. Industrial Partnerships
19.1. Company Names:
None at this time.
19.2. Rationale:
Not applicable
19.3. Contract Mechanism:
Companies are contacted by seminars and presentations.
19.4. Other Potential Companies:
The technology has been developed with university participants. Thus, many of the rights will belong to the universities. Many potential companies at international meetings.
19.5. International:
Presentations are at international meetings held in the US.
20. Intellectual Property
20.1. Patent Ownership:
None.
20.2. Other Owners:
20.3. Patent Number:
21. Cost Sharing:
No cost sharing is in place.
22. Background on this technology (Where did the idea come from? Who else is doing similar work? What have the results been to date? What is the most significant competitor to this technology?):
This technology builds on microbial monitoring that has gone on for 20+ years. New developments include the specific DNA probes applied, specific procedures developed, additional information on relevance of the PFLA data. University laboratories apply many of these techniques in collaboration with government and industry.
23. Reference Documents: