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Plant Growth Promoting Rihzobacteria
Enhanced Phytoremediation Systems for
  Remediation of Contaminated Soils

              Xiao-Dong Huang, Ph.D.
                  Vice President

     Waterloo Environmental Biotechnology Inc
                Waterloo, Ontario
Outline
1. Bacteria enhanced phytoremediation
3. Remediation cases: PHC and Salt
3. Cost analysis
Advantages of Phytoremediation

1.   Preserves natural environment and improves soil quality
2.   It is driven by solar energy and suitable to most regions
     and climates
3.   It is cost effective and technically feasible
4.   Restoration of ecosystem
5.   Can be used effectively at remote sites
6.   Effective for remediation of PHC and salt – relevant to the
     energy industry
Disadvantages of Phytoremediation


Phytoremediation process is too SLOW
10-20 years for completion
Feasibility ? Not good
The Question:
How to speed up the process to 10-20 months?
The answer:
Soil Rhizobacteria
Microorganisms in soil:
Plant associated bacteria:

Plant Growth Promoting
Rhizobacteria (PGPR)
Degrading Bacteria:

Contaminant degradation
Inoculation
to plants
1 week later




                  Inoculated
uninoculated
Greenhouse experiment for PHC degradation




                                                        Bacteria enhanced
                                    Bacteria enhanced
Bioremedaition   Phytoremediation                       Phytoremediaion
                                    Phytoremediaion
Greenhouse testing: PHC remediation




       phytorem             Enhanced phytorem
Plant promote rhizobacteria population


                                                               5 x 108
                                              Phytorem
      Rihzospere bacteria population, log
                                              Enhanced phyto
                                              Biorem
                                              有                5 x 107




                                            June        July     August   September
                                            seeding
Degradation kinetics of bacteria enhanced phytoremediation

                        60
                                                                  Phytoremediation
                                                                  Enhanced Phytorem
                        50
Degradation rate , %




                        40


                        30


                        20


                        10


                        0
                             0        2          4            6            8
                                                  time
HPLC analysis of PAH Degradation by PEP System

                    PYR               BBF
                                              BAP
                                CHR

                FLA                                             DBP
                          BBA               BKF           BGP

                                                    DBA               3 months
              PHE                                                     40%
Application of PEPS for PHC Remediation(2007- 09)

All sites met applicable criteria
1.   Sarnia, ON – land farm for oil sludge – 3 year study (15% w/w –
     60% F3 (C16-C34), 30% F4 (C34 – C50)
2.   Quebec City, QC – Tier 1 criteria met in one year
3.   Steinbach, MB – Complete remediation in 1 year – Gas station
     site
4.   Hinton, AB – Complete remediation in 2 years – Diesel invert
     drilling waste
5.   Peace River, AB – Complete remediation in 3 years – Flare pit
     material
6.   Edson, AB – Complete remediation in 2 years – Diesel invert
     drilling waste
Field application: Sarnia refinery landfarming site




                                                      2006/6/19
Field application: Sarnia refinery landfarming site



                          3 years remediation: PHC from 15% down to 3%
  PHC, %
28



                  21                                               35%




           g/kg
                  14



                  7




                       F2 (C16-15)   F3(C16-25)   F4 (C26-50)   >C50

Phytorem
                                                  PEPs
Quebec City, QC
PHC contaminated soil
treated with bioremediation for one year
Quebec City, QC – start of Season
              May 20 2009
                             Quebec tier 1 criteria: 300 ppm


 Soil contain 1000 ppm PHC




                                         Treatment pads
Quebec City, QC – End of Season
         Oct 20 2009
PHC Removal for Growth Season




                 QB Tier 1
                 standard
Field application: Gas Station in Steinbach, Manitoba

                  One year remediation:
          PHC from 2200 ppm down to 1000 ppm
Plant Growth and Coverage at a Diesel Invert Site near Hinton AB
                                      2008 remediation operation:
                                      Plating on June 25
                                      Final sampling on Oct 5
                                      Plant coverage 95%
                                      Pant height: 45 cm
PHC removal at Hinton site, AB




                       Alberta Tier 1
                       standard
A closed contaminated site near Peace River, AB
PHC Remediation at Peace River Site, AB




                                     28%
             Alberta Tier 1
             standard
                                      51%
Plant growth and coverage at a diesel invert site near Edson AB




                                                     July 31 2008
PHC Remediation near Edson, AB




                         Alberta Tier 1
                         standard
Second Generation Full Scale Sites

1.   Three sites near Dawson Creek, BC
2.   One site near Fort Nelson, BC
3.   One site near Swan Hills, AB
4.   One site near Hinton, AB
5.   One site near Edson, AB
6.   One site near Red Earth Creek, AB
Application of PEPS for Salt Remediation
                (2007- 09)

        1.   Norman Wells, NWT
        2.   Provost, AB
        3.   Cannington Manor, SK
        4.   Alameda, SK
        5.   Kindersley, SK
        6.   Brazeau, AB
        7.   Weyburn, SK (7 sites)
        8.   Red Earth, AB
Norman Wells, NWT – Start of Season (2008)
Soil Impact – Salt
Norman Wells, NWT – End of Season (2010)
Soil Impact – Salt
Norman Wells, NWT – End of Season (2009)
Soil Impact – Salt




                       Plant Biomass (dry wt g/m2 ± SE)
                     Year    Plot A     Plot B     Plot C
                     2009   300 ± 26   397 ± 50   623 ± 44
                     2010   393 ± 16   592 ± 40   525 ± 20
Provost, AB: End of Season (2009, Year 1)
Soil Impact – Salt
 •
     High salt spots have filled in with plants
                                                  •
                                                      ECe (2009):
                                                      decreased from
                                                      13-17 to 4-12
                                                      dS/m
                                                  •
                                                      ECe (Spring
                                                      2010): all
                                                      sampling points
                                                      were below
                                                      applicable targets
                                                  •
                                                      Successful
                                                      remediation was
                                                      achieved in 1
                                                      year
Phytoremediation Cost analysis for the Edson Site

•
    1.07 ha impacted to a depth of 0.3 m or 3,210 m3 of PHC
    impacted material
•
    The costs for the entire project for PEPs was: $104,000 or
    $32.50/m3
•
    Landfilling this material would have cost $70/m3
     •
       Assumes a 2 h truck turnaround time if no backfill
       required
     •
       If backfill was required, the cost would rise to $90/m3
•
    Composting would cost $75-150/m3
Examples of Remediation Methods
•   Dig and dump - Any contaminant type - $100-500/m3
•   Soil incineration - On or off site - Organic contamination -
    $200-600/m3
•   Chemical extraction - Any type of contamination -
    $250/m3
•   Electrokinetic separation - Metals/Salts - $200/m3
•   Soil flushing/fracturing - Any contaminant type - $250/m3
•   Land farming - Small organics - $50/m3
•   Bioremediation - Organics - $100/m3
•   Our PEPs - Any contaminant type - $25-50/m3
Development, Proof, and Application of
        PGPR Enhanced Phytoremediation
               Systems (PEPS)
Over 10 years of research and full-scale field
application:
1.   PHC: 18 sites in AB, BC, MB, QC and ON(2004-10)
2.   Gas station: site fully remediated in 1 summer (2007)
3.   Salt: 14 sites in SK, AB and NWT (2007-10)
Why Use Phytoremediation?
•
    Works effective for PHC and salt remediation.
•
    Remediation at all sites have been successful; > 20 sites.
•
    Costs of PHC and salt remediation are low.
•
    Unit cost drops as the volume of material increases.
•
    Phytoremediation costs (all in) < half the cost of landfilling.
•
    Liability is reduced, not transferred to a landfill.
•
    Cost effective at remote sites.
•
    Purchase of backfill not required. Soils are reused.
•
    Tier 2 approach - if required only marginal cost increase.
•
    Green technology, solar energy and carbon sequestration.
•
    Environmentally friendly and Positive PR opportunity.
Colleagues and Partners
People in the group       Partners
   •
       Bruce Greenberg    •
                              S Willets, O Mrklas, C Gordey,
   •
       Karen Gerhardt             Conoco Phillips Canada
   •
       Jola Gurska
                          •
                              B Moore, Devon Canada Inc
   •
       Xiao-Ming Yu
                          •
                              E Harrison, Cenovus Energy
   •
       Mark Lampi         •
                              P Coldham, Questerre Engegy
   •
       Shan Shan Wu       •
                              L Lawlor, Imperial Oil
   •
       Julie Nykamp       •
                              A Traverse, Baytex Energy Corp
   •
       Nicole Knezevich   •
                              G Millard, Shell Canada
   •
       Greg MacNeill      •
                              Canadian Forestry Oil Corp
   •
       Xiaobo Lu          •
                              Perry Gerwing, Earthmaster
   •
       Scott Liddycoat
                          •
                              T Chidlaw, MWH
   •
       Han Zheng
   •
       Brianne McCallum
                          •
                              J Budziak, Seaway
   •
       Peter Mosley       •
                              D McMillan, SNC
   •
       Matt Hannaberg     •
                              G Stephenson, Stantec
                          •
                              S Steed, NorthWind
                          •
                              G Adams, URS

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ONEIA Xiao-Dong Huang presentation

  • 1. Plant Growth Promoting Rihzobacteria Enhanced Phytoremediation Systems for Remediation of Contaminated Soils Xiao-Dong Huang, Ph.D. Vice President Waterloo Environmental Biotechnology Inc Waterloo, Ontario
  • 2. Outline 1. Bacteria enhanced phytoremediation 3. Remediation cases: PHC and Salt 3. Cost analysis
  • 3. Advantages of Phytoremediation 1. Preserves natural environment and improves soil quality 2. It is driven by solar energy and suitable to most regions and climates 3. It is cost effective and technically feasible 4. Restoration of ecosystem 5. Can be used effectively at remote sites 6. Effective for remediation of PHC and salt – relevant to the energy industry
  • 4. Disadvantages of Phytoremediation Phytoremediation process is too SLOW 10-20 years for completion Feasibility ? Not good The Question: How to speed up the process to 10-20 months? The answer: Soil Rhizobacteria
  • 5. Microorganisms in soil: Plant associated bacteria: Plant Growth Promoting Rhizobacteria (PGPR) Degrading Bacteria: Contaminant degradation
  • 7. 1 week later Inoculated uninoculated
  • 8.
  • 9. Greenhouse experiment for PHC degradation Bacteria enhanced Bacteria enhanced Bioremedaition Phytoremediation Phytoremediaion Phytoremediaion
  • 10. Greenhouse testing: PHC remediation phytorem Enhanced phytorem
  • 11. Plant promote rhizobacteria population 5 x 108 Phytorem Rihzospere bacteria population, log Enhanced phyto Biorem 有 5 x 107 June July August September seeding
  • 12. Degradation kinetics of bacteria enhanced phytoremediation 60 Phytoremediation Enhanced Phytorem 50 Degradation rate , % 40 30 20 10 0 0 2 4 6 8 time
  • 13. HPLC analysis of PAH Degradation by PEP System PYR BBF BAP CHR FLA DBP BBA BKF BGP DBA 3 months PHE 40%
  • 14. Application of PEPS for PHC Remediation(2007- 09) All sites met applicable criteria 1. Sarnia, ON – land farm for oil sludge – 3 year study (15% w/w – 60% F3 (C16-C34), 30% F4 (C34 – C50) 2. Quebec City, QC – Tier 1 criteria met in one year 3. Steinbach, MB – Complete remediation in 1 year – Gas station site 4. Hinton, AB – Complete remediation in 2 years – Diesel invert drilling waste 5. Peace River, AB – Complete remediation in 3 years – Flare pit material 6. Edson, AB – Complete remediation in 2 years – Diesel invert drilling waste
  • 15. Field application: Sarnia refinery landfarming site 2006/6/19
  • 16. Field application: Sarnia refinery landfarming site 3 years remediation: PHC from 15% down to 3% PHC, %
  • 17. 28 21 35% g/kg 14 7 F2 (C16-15) F3(C16-25) F4 (C26-50) >C50 Phytorem PEPs
  • 18. Quebec City, QC PHC contaminated soil treated with bioremediation for one year
  • 19. Quebec City, QC – start of Season May 20 2009 Quebec tier 1 criteria: 300 ppm Soil contain 1000 ppm PHC Treatment pads
  • 20. Quebec City, QC – End of Season Oct 20 2009
  • 21. PHC Removal for Growth Season QB Tier 1 standard
  • 22. Field application: Gas Station in Steinbach, Manitoba One year remediation: PHC from 2200 ppm down to 1000 ppm
  • 23. Plant Growth and Coverage at a Diesel Invert Site near Hinton AB 2008 remediation operation: Plating on June 25 Final sampling on Oct 5 Plant coverage 95% Pant height: 45 cm
  • 24. PHC removal at Hinton site, AB Alberta Tier 1 standard
  • 25. A closed contaminated site near Peace River, AB
  • 26. PHC Remediation at Peace River Site, AB 28% Alberta Tier 1 standard 51%
  • 27. Plant growth and coverage at a diesel invert site near Edson AB July 31 2008
  • 28. PHC Remediation near Edson, AB Alberta Tier 1 standard
  • 29. Second Generation Full Scale Sites 1. Three sites near Dawson Creek, BC 2. One site near Fort Nelson, BC 3. One site near Swan Hills, AB 4. One site near Hinton, AB 5. One site near Edson, AB 6. One site near Red Earth Creek, AB
  • 30. Application of PEPS for Salt Remediation (2007- 09) 1. Norman Wells, NWT 2. Provost, AB 3. Cannington Manor, SK 4. Alameda, SK 5. Kindersley, SK 6. Brazeau, AB 7. Weyburn, SK (7 sites) 8. Red Earth, AB
  • 31. Norman Wells, NWT – Start of Season (2008) Soil Impact – Salt
  • 32. Norman Wells, NWT – End of Season (2010) Soil Impact – Salt
  • 33. Norman Wells, NWT – End of Season (2009) Soil Impact – Salt Plant Biomass (dry wt g/m2 ± SE) Year Plot A Plot B Plot C 2009 300 ± 26 397 ± 50 623 ± 44 2010 393 ± 16 592 ± 40 525 ± 20
  • 34.
  • 35. Provost, AB: End of Season (2009, Year 1) Soil Impact – Salt • High salt spots have filled in with plants • ECe (2009): decreased from 13-17 to 4-12 dS/m • ECe (Spring 2010): all sampling points were below applicable targets • Successful remediation was achieved in 1 year
  • 36. Phytoremediation Cost analysis for the Edson Site • 1.07 ha impacted to a depth of 0.3 m or 3,210 m3 of PHC impacted material • The costs for the entire project for PEPs was: $104,000 or $32.50/m3 • Landfilling this material would have cost $70/m3 • Assumes a 2 h truck turnaround time if no backfill required • If backfill was required, the cost would rise to $90/m3 • Composting would cost $75-150/m3
  • 37. Examples of Remediation Methods • Dig and dump - Any contaminant type - $100-500/m3 • Soil incineration - On or off site - Organic contamination - $200-600/m3 • Chemical extraction - Any type of contamination - $250/m3 • Electrokinetic separation - Metals/Salts - $200/m3 • Soil flushing/fracturing - Any contaminant type - $250/m3 • Land farming - Small organics - $50/m3 • Bioremediation - Organics - $100/m3 • Our PEPs - Any contaminant type - $25-50/m3
  • 38. Development, Proof, and Application of PGPR Enhanced Phytoremediation Systems (PEPS) Over 10 years of research and full-scale field application: 1. PHC: 18 sites in AB, BC, MB, QC and ON(2004-10) 2. Gas station: site fully remediated in 1 summer (2007) 3. Salt: 14 sites in SK, AB and NWT (2007-10)
  • 39. Why Use Phytoremediation? • Works effective for PHC and salt remediation. • Remediation at all sites have been successful; > 20 sites. • Costs of PHC and salt remediation are low. • Unit cost drops as the volume of material increases. • Phytoremediation costs (all in) < half the cost of landfilling. • Liability is reduced, not transferred to a landfill. • Cost effective at remote sites. • Purchase of backfill not required. Soils are reused. • Tier 2 approach - if required only marginal cost increase. • Green technology, solar energy and carbon sequestration. • Environmentally friendly and Positive PR opportunity.
  • 40. Colleagues and Partners People in the group Partners • Bruce Greenberg • S Willets, O Mrklas, C Gordey, • Karen Gerhardt Conoco Phillips Canada • Jola Gurska • B Moore, Devon Canada Inc • Xiao-Ming Yu • E Harrison, Cenovus Energy • Mark Lampi • P Coldham, Questerre Engegy • Shan Shan Wu • L Lawlor, Imperial Oil • Julie Nykamp • A Traverse, Baytex Energy Corp • Nicole Knezevich • G Millard, Shell Canada • Greg MacNeill • Canadian Forestry Oil Corp • Xiaobo Lu • Perry Gerwing, Earthmaster • Scott Liddycoat • T Chidlaw, MWH • Han Zheng • Brianne McCallum • J Budziak, Seaway • Peter Mosley • D McMillan, SNC • Matt Hannaberg • G Stephenson, Stantec • S Steed, NorthWind • G Adams, URS