bioremediation of groundwater and soil contaminated with
TRANSCRIPT
Bioremediation of Groundwater and Soil Contaminated with Hazardous Chemicals
2006.5.17
Osami Yagi
Advanced Research Institute for the Sciences and HumanitiesNihon University
1. Current status of groundwater and soilpollution
2. Guidance to bioremediation technologies
3. Field application of bioremediation
4. Ongoing research on bioremediation
5. Future aspects of bioremediation
Outline
Groundwater and Soil Pollution in JapanGroundwater and Soil Pollution in Japan
Use of GW Living 21％, Industrial 26％
GW Pollution Cumulative numbers (2004) 3,120 Tetrachloroethylene 632Trichloroethylene 480NO2-N, NO3-N 1,398
Soil Pollution Japan (2003) 1,458US 217,000Germany 300,000Holland 500,000
Survey of Groundwater Pollution (2004)
Substance Surveyed wells
Pollutedwells
Ratio(%)
lead 14
cis-1,2-dichloroethylene 5trichloroethylene 18
tetrachloroethylene 22
total 4,955 387 7.8
NO2-N, NO3-N 235FB
19
4,234
4,2484,2603,5423,499 8
0.40.55.50.50.2
3,566
3,743
0.4
total mercury 53,235 0.2
1,1-dichloroethylene 23,744 0.10.1
arsenic 3,666 74 2.0
carbontetrachloride 43,661 0.1
Chemicals
Conventional remediation technologies for groundwater and soil
Reduce flux to groundwater
Reduce flux to groundwater
Removal
Technology Mechanism of treatment
Heavy metals
Halogenated compounds
Solidification/ Stabilization
Sealing
Excavation
Soil vapor extractionExcavation
Bioremediation
Pumping and treatment
Removal
Removal
Removal
Degradation
Ongoing New Remediation Technologies for VOC
Technology Mechanism of treatment
Physical and chemical
・Chemical oxidation Degradation
・Zero valent Iron Reductive dehalogenation・Thermal treatment Heating of soil/volatilization
・Surfactant flush and solvent extraction
Extraction
H2O2, KMnO4
Bioremediation
Bioremediation
Bioremediation is an engineered process that uses microorganisms to clean up the polluted environment by hazardous chemicals.
Approach
BiostimulationThe addition of organic or inorganic compounds to
cause indigenous organisms to effect remediation of the environment, e.g. nutrients, lactate, CH4
BioaugmentationThe addition of microorganisms to effect remediation
of the environment, e. g. contaminant-degrading bacterial injection into an aquifer
Definition
Current and Future Application of Bioremediation
Substrates Water Air Wastewater
Heavy metalsHg P MCd PCr6+ M
Toxic chemicalsPCB _ MTCE MPCE MDioxin PEndocrine disruptor P MNOx, SOx P
Other chemicalsBOD, COD P MNO3-N, NH4-N P MP P MOil M
__ _
_ _
__ __ _
_ __
_ _
__
Soil
MP
MMMMM
M, P
M, P
_
_
_
_ __ M
M: Microbes P: Plant : Practical use
Ex situBiopileLand treatmentSlurry phase treatmentBioreactor
In situBioventingBiospargingDirect injectionPumping and treatment, RecirculationPermeable Barriers, Permeable reactive wall
Bioremediation Technologies
Biopile
Offgas Treatment Blower
Liner Perforated laterals
Vaper Cover Optional
Pipe
Bioventing
Extraction well
Contamination
Air flow
Water table
Injection well
Air
Injection well
Air
・Light oil, sandy soil
Biosparging
・Vapor extraction and biodegradation
Ｗ．Ｌ
Oil pollution
Unsaturate
Saturate
Air
Gas
Unpermiable zone
AC carbon
RoadWall Extpump
Comp
Separate
Air N, P, CH4, Microbes
Vadose zone
Water Table
Aquifer
Plume
In situ bioremediation
Extraction well
Permeable Barriers or Zones
• Iron walls - biofilms, • Biosparge/Biofilter walls - secondary effects of redox and
pH increases
Factors for BioremediationFactors for Bioremediation
Type Pollutant Electron donor Electron acceptor oil
TCE(PCB)(dioxin)PAH
CH4、C3H8、Toluene, Phenol
PCETCEdioxin
NO3-N
N, P, Microorganisms
Aerobic O2, H2O2CaO2, MgO2
Anaerobic Lactate, Poly-Lactate, Acetate, Ethanol, H2
Soil Reduction method (Hybrid process)– Mechanism
•Reductive Dechlorination (Chemical reaction, Iron) •Anaerobic Dechlorination (Microbial reaction, Organic acid)
RCl + H+
RH + Cl-
e-M+
H2O
OH- + H2
M++ RCl + H+
RH + Cl-e-
M+
RCl
RH + Cl- + H++Cat
M0
M0
Reduction by Iron
RCl
H2
FermentationElectron-donor
Electron acceptor
Reduction by anaerobic bacteria
RH + Cl- + H+
Biomass
Organic
Inter-action
？
？
Re
Re
Matheson & Tratnyek, 1994
Proposed Pathways for the Dechlorination of PCE under Anaerobic Condition
C CCl
Cl
Cl
Cl
C CCl
H
Cl
Cl
C C
Cl
H
Cl
H
C CH
Cl
Cl
H
C C
H
H
Cl
Cl
C CH
H
Cl
H
C CH
H
H
H
H2 HCl H2 HCl H2 HCl H2 HCl
PCE TCE
Trans-
DCE
Cis-
1.1-
VC ETH
• Field application for VOC pollution–Pollutant
•TCE and cDCE•Max con： 21 mg/L（TCE）•Groundwater 15 mg/L（cDCE）
– Area•Depth： （Max depth 20m）
•Unsaturated + saturated•Volume： 100,000 m3
– Site•Building removed place
• Treatability test• Step 1： gene diagnosis → negative（D. ethenogens not present）• Step 2： laboratory test → degradation
• Method• Iron and organic acid injection• Unsaturated zone and high concentration saturated zone, Mixing• Groundwater circulation
• Cis-DCE concentration
Before Mixing areaInjection area
Mixingafter 2 months
Mixing after 4 months
Characteristics of TCE
M. W. 131Removal of fat and solventHepatotoxic and carcinogenic
Henry‘ s const. 11.7 l ・atm/ mol •
CCl
HCl
ClC
TCE degradation (%) by Methylocystis sp. M
Conditions TCE concentration (mg/l)
Growing cell*1
Resting cell*2
Immobilized cell*2
190
90
90
3.590
50
50
1055
25
30
350
5
10
100-
0
0
*1 Incubated for 15days*2 Incubated for 10hr
Thin-section electron micrograph of Methylocystis sp. M
0.5mm
TCE degradation pathway by strain M
TCE
CO2
Methane monooxygenase
CH4 CH3OH
NADH NAD+
O2 H2O
Chloral
TCE oxide intermediate
Cl C
Cl
Cl
CH
O
ClC
ClClC
O
H
ClC
Cl
ClC
H
Trichloroacetic acid
1,1,1-Trichloroethanol
Cardonmonooxide Formic acid Glyoxlic acid Dichloroacetic acid
15 cm 15 cm
150 cm
50 cm
50 cm
170 cm
PP
TCE contaminationg soil
PPCH4, O2, P, N
strain M
Sandy soil lysimeter
1050-5-100.0
0.1
0.2
Days
TC
E (m
g/l)
control
strain M
with strain MTCE degradation in sandy soil lysimeter
DioxinsO
O O
Dibenzo-p-dioxin (DD)
Dibenzofuran (DF)
Coplanar-PCB (Co-PCB)
Environmental standardSoil: 1000pg-TEQ/g, Water : 1pg-TEQ/L
Dioxin emissionsMore than 80 % from incineration facilities
Polluted site Nose,Osaka: 8500 pg-TEQ/g (1998)Omori,Tokyo: 16,000 pg-TEQ/g (2000)
Pollution in Japan
Shape Short-rods(0.8×1.0 μm)
Gram stain +Spore formation -Motility -Colonies Circularon Nutrient Agar Entire margin
48 h incubation ConvexGlistenYellow
Growth at 37 °C +Growth at 45 °C +w
Growth underNaCl
10.0 % +
DF-utilizing bacterium, strain YA
500nm
Electron micrograph of
Strain YA
Degradation of 1,2,3,4Degradation of 1,2,3,4--TT44ClDD by Resting CellsClDD by Resting Cells
O
O
Cl
ClCl
Cl
•109cells/mL•1,2,3,4-T4ClDD : 10mg/L, 2wk, triplicate•GC-MS
0
2
4
6
8
10
12
Cont AS1 AS2 AS3 AS4 AS5 YA YZ JTStrain
mg/l
iter
0
2
4
6
8
10
12
Cont AS1 AS2 AS3 AS4 AS5 YA YZ JTStrain
mg/l
iter
Hg
Hg
Hg
HgHg Hg
Hg
Hg
Hg
Hg
Hg
Hg
Mercury-contaminated areas
Hg
Hghuman exposureaquatic organism exposure
inorganic mercuryorganomercury compounds
Ghana
Pakistan
Iraq
Switzerland
Sweden Finland China
Greenland
Japan
Brazil
Venezuela
Guatemala
USA
Canada
Hg
Niigata Minamata
Hg Hg
Pseudomonas putida PpY101/pSR134
NADPH NADP+
02+Hg
merR
merT
merA
transfer protein
mercury reductase
pSR134
mer operon
Hg
Rem
oval
Hg2
+(%
)
0
20
40
60
80
100
NaCl 0M
NaCl 0.02M
NaCl 0.2M
control(without cell)
Removal of mercury from soil slurry by P. putida PpY101/pSR134
Thiol 1 mMCells 100 mg dry weight (4.0 x 108 CFU/ml)
Soil sample 20 g dry weight
0, 0.02, 0.2 MNaClHgCl2 40 mg-Hg/LDistilled water 200 ml
Reaction condition
30ÞC, 120rpm, 23hr
Brassica juncea2,080~8,240 mg-lead /kg plant
Lead Phytoremediation (Kondo etc. 2002)
PhytoremediationPhytoremediationArsenic (Arsenic (PterisPteris vittatavittata ））
• Dr. Ma• Hyper accumulater• Accumulation 2.2％• Biomass 2～3kg/㎡ yr
Clean up by self-purification
It is very usefulto predict the clean up periodto select the bioremediation technology
How to determine the natural attenuation biological degradation rate, non-biodegradation rate
diffusion, dispersion, adsorption, volatilization
Factorspollutant, pollution concentration,organic concentration, DO, pH, ORP, temperature
Natural Attenuation
Future aspects of bioremediation technologies
1. Enhancement of natural attenuationMonitoring microbial community
2. Development of in situ bioaugmentationIncrease the microbial activity
3. Development of hybrid remediation technologies
4. Phytoremediation
Aerobic and anaerobic, Chemical and biological, Physical and biological
Establish toxic chemical accumulating and degrading plant
5. Public acceptanceUnderstand the new technology