in situ bioreactors for in-well groundwater remediation · the bio-sep isbr • bio-sep beads...
TRANSCRIPT
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In Situ Bioreactors for In-Well Groundwater Remediation
Eric J. Raes, P.E., LSRPKerry Sublette
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The Bio-Sep In Situ Bioreactor (ISBR)
• Enhancement of in situ bioremediation in groundwater with compact bioreactor installed in-well
• Overcomes common limitations of bioremediation of groundwater– Low contaminant concentrations
• A threshold concentration of substrate is required for growth
– Substrate inhibition• At high concentrations some biodegradable
contaminants can be toxic to the organisms that have the ability to degrade them
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In the beginning…..
The Bio-Trap® Sampler:• Rapidly colonized by indigenous
bacteria forming active biofilms• Thousands used worldwide for
over a decade for forensic analysis of groundwater microbiology
Cell division inside of Bio-Sep® Bead within a Bio-Trap Sampler
X-Section of Bio-Sep® Bead Interior of Bio-Sep® Bead
Bio-Trap® Sampler with Bio-Sep® Beads
Nomexand PAC
• Adsorptive surface• High surface area
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The Bio-Sep ISBR• Bio-Sep beads provide an incredible
surface area for microbial growth• Gas sparging (air or N2) creates an
airlift for circulation of groundwater through the bioreactor. • Contaminated groundwater is
treated as it moves through the column of Bio-Sep beads
• Nutrient addition (N, P, electron donors, electron acceptors) support growth of desired indigenous microbes
• Water exiting the reactor carries contaminant-degrading microbes into the aquifer
1 ¼” PVCFits in 2” well
Air or N2 and nutrient delivery
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Topside control
• Nutrient reservoirs and pumps
• Air pump• Air flow control
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• Petroleum hydrocarbons• Aerobic• Anaerobic
• Chlorinated hydrocarbons• Anaerobic
• Fuel oxygenates (MTBE, TBA)• Emerging contaminants (1,4-Dioxane)
Bio-Sep ISBR Applications
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Case Study – Aerobic ISBR – Low Concentrations of Hydrocarbons• Fuel oil release impacting soil and groundwater beneath
a private residence• The bioreactor well & four monitoring wells were
installed in the basement• Wells spaced 1 ft. apart
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ISBR Timeline
• Days -67 through -34: Air sparging only (no Bio-Sep)
• Days -34 through 0: Air sparging and nutrient delivery (no Bio-Sep)
• Day 0 onward: Complete bioreactor system operational (Bio-Sep beads added)
• Bio-traps® (Microbial Insights, Inc) used through out testing to monitor microbiology of bioreactor well and monitoring wells
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qPCR Analysis of BR1 Bio-traps During ISBR Testing
Didn’t see much difference in hydrocarbon catabolic genes with DNA. This is common observation since hydrocarbon degraders are ubiquitous in the environment.
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RT-qPCR Analysis of BR1 Bio-traps During ISBR Testing
No expression of hydrocarbon catabolic genes until the complete bioreactor system was in service.
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Anaerobic ISBRs, the same but different
N2 sparger
1 ¼” PVCFits in 2” well
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Case Study – Anaerobic ISBR
• Chlorinated solvent impacted site
• Fractured bedrock aquifer
• Deep groundwater impacts (140’ bgs)
• Unfavorable geochemistry
• Low but measureable DO
• DO increase with rain event
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Case Study – Anaerobic Bioreactor
• ISBR installed at a depth of 30’ BGS• “Liquid carbon” electron donor• Groundwater monitoring
• Contaminant concentrations
• Geochemistry
• qPCR for Dehalococcoides and functional genes for reductive dechlorination (bio-traps and groundwater)
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Groundwater Contaminants & Microbiology
0
200
400
600
800
1000
1200
1400
1.0E+00
1.0E+01
1.0E+02
1.0E+03
1.0E+04
1.0E+05
1.0E+06
0 1 2 3 4 5
Conc
entr
atio
n (µ
g/L)
Cells
/mL
DHC BVC VCR DHBt PCE TCE cDCE Ethene VC
Quarters
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Dehalococcoides (DHC)Concentration with Depth
(Monitored by Bio-traps)
1.00E+00 1.00E+01 1.00E+02 1.00E+03 1.00E+04
-140
-105
-85
-60
Cells or gene copies/bead
Elev
atio
n (ft
)
ND
1.00E+00 1.00E+01 1.00E+02 1.00E+03 1.00E+04 1.00E+05
-140
-105
-85
-60
Cells or gene copies/bead
Elev
atio
n (ft
)
Pre-ISBR Post-ISBR
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1.00E+00 1.00E+01 1.00E+02 1.00E+03 1.00E+04
-140
-105
-85
-60
Cells or gene copies/bead
Elev
atio
n (ft
)
Vinyl Chloride RDases with DepthPost-ISBR
1.00E+00 1.00E+01 1.00E+02 1.00E+03 1.00E+04
-140
-105
-85
-60
Cells or gene copies/bead
Elev
atio
n (ft
)
ND
NDND
NDND
NDND
Pre-ISBR
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ISBRs Will Transfer Degraders from One Well to Another
In situtreatment and colonization of ISBR
Inoculation and/or kick start degradation
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Transfer of DHC in a TCE Plume
0
500
1000
1500
2000
2500
3000-9
00
-870
-840
-810
-720
-630
-540
-450
-360
-270
-180 -90 0 30 60 90 120
150
Time (days)
[VO
C],
ug/L TCE
cis-DCEVCEthene
nZVI/
Emulsified oil
Colonized ISBR installed
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When to consider an ISBR
• Inhibitory contaminant concentrations
• Dilute plumes (persistent low levels of contaminants)
• Following ISCO
• Difficult situations
• Limited physical access
• Where one-time amendment injection is not feasible
• Where bioremediation has failed previously
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ISBR Limitations
• Aerobic operation limited to low concentrations of reduced iron (fouling)
• Radius of influence decreases with increasing hydraulic conductivity of aquifer matrix
• Works best with contaminants adsorbed by activated carbon
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ISBR O&M and Costs
• O&M
• System checks every 2-4 wks
• Power
• Nutrients
• Water level (ISBR must be totally submerged to function)
• Costs
• Life of project rental
• $10,000 for one unit (ISBR and controller)
• $15,000 for two units
• Decreasing per unit costs with addition of more units at a given site
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Eric RaesBioEnhance, [email protected](973)219-6185
Kerry SubletteBioEnhance, [email protected](918)691-0639
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Chemical Oxidation & Aerobic ISBR
Case study: Fuel oil release
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Naphthalene degraders Pre- and Post-ISCO
0
2000
4000
6000
8000
10000
1.0E+00
1.0E+02
1.0E+04
1.0E+06
1.0E+08
1.0E+10
Pre-ISCO ISCO Post-ISCOBiostimulation
Post-ISCOMNA
δ13 C
DIC
(‰)
Gen
e Co
pies
/bea
d
NAH Population (DNA) NAH Expression (RNA) DIC Del (‰)
Present but not active
Present & active Aerobic naphthalene
degraders are present but not
active
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0
2000
4000
6000
8000
10000
1.0E+00
1.0E+02
1.0E+04
1.0E+06
1.0E+08
δ13 C
DIC
(‰)
Gen
e Co
pies
/bea
d
NAH Population (DNA) NAH Expression (RNA) DIC Del (‰)
ND NA
Naphthalene degraders ISBR – Post-ISCO
No ISBRPresent but not active
ISBR DeactivatedNo activity or mineralization
ISBR InstalledActive &
mineralizing
ISBR Re-installedActive &
mineralizing