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1. Bioremediation-From the Lab to the Field
Mitch Lasat, Ph.D.
NCER/ORD
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2. •What is Bioremediation?
•Why Bioremediation works?
•Contaminants amenable to Bioremediation
•Limiting factors (why bioremediation doesn’t work?)
•Engineering strategies for Bioremediation
•Is bioremediation a “hot” research topic for the EPA?
•Bioremediation research
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Presentation Outline

3. What is Bioremediation?
 the use of biota to degrade/mitigate environmental contamination
-bioremediation- by microorganisms (soil, groundwater-organic
contaminants)
-phytoremediation- by plants (mostly soil and surface water)
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4. Why Bioremediation works?
 microorganisms obtain energy for growth by degrading organic contaminants
in an enzyme-mediated process- direct metabolism
-aerobic biodegradation of BTEX in the presence of an oxygenase
(Pseudomonas)
 some enzymes are not very specific and in addition to the growth substrate
transform other compounds-cometabolism
-oxygenases are not very substrate-specific and can also degrade TCE
(however TCE cannot be used as a growth substrate)
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5. Contaminants amenable to Bioremediation I
 Hydrocarbons:
- BTEX (aerobic and anaerobic biodegradation)
- PAH (less amenable)
- aerobic degradation via cometabolism
- anaerobic biodegradation (naphtalene-denitrification)
 Chlorinated Aliphatic Hydrocarbons
- aerobic electron donor (DCM, CM, DCA)
- anaerobic electron donor (TCE, DCE)
- anaerobic acceptor (PCE, TCE)- dehalorespiration
- cometabolism (aerobic, anaerobic-reductive dechlorination)
 Chlorinated aromatic hydrocarbons
- PCB (in general bioremediation recalcitrant)
-aerobic, less chlorinated
-anaerobic (dehalorespiration)
- PCP; aerobic, anaerobic (groundwater-reductive dechlorination)
- Dioxins; highly resistant to Bioremediation
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6. Contaminants amenable to Bioremediation II
Pesticides
-chlorinated; highly resistant to aerobic transformation
-phosphorus based and carbamate; quickly hydrolyzed
-triazine; biodegradable
Explosives
-biotransformation is partial (TNT) or slow (RDX)
Inorganics
-bacterial reduction of Hg2+ to Hg0
-bacterial reduction of Cr6+ to Cr3+
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7. Factors that limit the potential for Bioremediation
(why Bioremediation doesn’t work)
1) Contaminant-related limitations:
 Synthetic vs. natural contaminants
-bioremediation potential greater for natural compounds
 Physical characteristics
-density, Henry’s constant, solubility, octanol/water partition coefficient
 Molecular structure of the contaminant
-extent of chlorination, linear vs. branched structure, saturated vs.
unsaturated compounds
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8. Factors that limit the potential for Bioremediation
(why Bioremediation doesn’t work)
2) Environmental conditions:
Hydrogeology: permeability/hydraulic conductivity, heterogeneity,
fracture bed rocks, soil properties, pH
 Nutrients: C:N:P-100:10:1
Electron acceptor: oxygen (3 parts of oxygen to converts 1 part of
hydrocarbon to CO2), nitrate, sulfate, ferric iron
3) Microorganisms presence:
Assessment of microbial activity, introduced microorganisms
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9. Engineering Strategies for Bioremediation
Intrinsic bioremediation/natural attenuation
Enhanced/engineered bioremediation
-addition of nutrients, oxygen
Bioaugmentation
-introduction of appropriate organisms
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10. Phytoremediation
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 Phytoextraction (removal/extraction of toxic metals- Pb)
 Phytodegradation (organics degradation in roots and shoots-
TPH, PAHs, BTEX, pesticides, CAHs)
 Phytovolatilization (CAHs, Hg, Se)
 Evapotranspiration/Hydraulic control (plume reduction)

11. Is bioremediation a “hot” research topic for the EPA?
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 Cross-agency research advisory workgroup for Goal 3; Bioremediation- a
priority technology for remediation of contaminated sediments,
groundwater and soil
 ORD GOAL 3 MYP-long-term goal oriented with annual progress measured
by completion of APG/APM-of the approximately 70 remediation-related
APMs, approximately half pertain to bioremediation:
- Report on biodegradation of PAHs in sediments
- Report on solvent-enhanced residual biotreatment of residual DNAPL
- Develop and evaluate microbial populations for effective TCE
biodegradation
- Develop and evaluate cost-effective methods for nutrient mixing and
delivery for bioremediation of chlorinated aliphatic hydrocarbons
- Synthesis report on 5 DNAPL remediation technologies

12. •Bioremediation research I
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 ORD’s Goal 3research program is designed to provide a better understanding
of the traditional risk management options (dredging, capping, pump and
treat), and to investigate alternative options (bioremediation, MNA)
 Problem-driven research program, supporting research needs of:
- Office of Solid Waste
- Superfund
- Leaking Underground Storage Tank Corrective Action
- Oil Spills
 Contacts: NPD-Randy Wentsel
NRMRL-Trish Erickson
NCER-Mitch Lasat

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 NCER Bioremediation research:
- 1997-2001; several RFAs on Bioremediation and Phytoremediation
- 2001; HSRC program was recompeted, research focus on contaminated
sediments, VOC-contaminated groundwater, mine wastes,
phytoremediation
http://es.epa.gov/ncer/grants/
 Case studies/performance data:
http://clu-in.org/techfocus/
- site general information
- contaminants
- site hydrology
- media
- cleanup goals
- technology used
- results/costs
- lessons learned
•Bioremediation research II