The Chemistry and Biology BehindSuccessful Phytoremediation of Petroleum        and Salt Impacted Soils Bruce Greenberg, X...
The Phytoremediation Process
Phytoremediation                         • Volatilization                         • Phytodegradation                      ...
Advantages of Phytoremediation 1. Soil quality improved 2. Driven by solar energy - suitable to most regions    and climat...
Development, Proof and Full Scale Application ofPGPR Enhanced Phytoremediation Systems (PEPS) Over 13 years of research wi...
The key to phytoremediation success -Transfer of the science from the lab to the field1. Strategies for aggressive plant g...
WEBi-Earthmaster-UW Partnership• Synergistic expertise in contaminated site remediation• Developed commercial phytoremedia...
PGPR Enhanced Phytoremediation Systems (PEPS)Aggressive plant growth strategies leads to remediation   Physical soil treat...
"So, what have we beendoing for the last year?"
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
Interaction of a PGPR Containing ACCDeaminase with a Plant Seed or Root           Amino           Acids                   ...
PGPR Isolation                 Plant Growth-Promoting                 Rhizobacteria (PGPR) Isolation                 • Hav...
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
PGPR Regeneration & Validation                    Regeneration of proven                    PGPR isolates for field use   ...
PGPR Validation – Assess plant growth    enhancement•   Before use in PEPS: plant growth assays to assure PGPRs perform pr...
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
Seed Treating & Shipping                           • Treat seeds with proven                             and regenerated P...
PGPR Seed Treatment QA/QCAliquots of PGPR-treated seeds assayed for plant growth enhancementto assure successful PGPR appl...
Seed Treating & Shipping                           • Treated seeds shipped to                             sites after QA/QC
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
Disking/Tilling to prepare seed bed
Application of appropriate fertilizer
Sample soil for beginning of season PHC and/or Salt levels
Sow PGPR-treated seed
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
Edson, AB – Before site prep and seeding All previous steps assure sites that looked like this…..                  Soil Im...
PEPS Deployment at Edson, AB…..Look like this               Soil Impact – PHC (Diesel Invert; 85% F3)
Weyburn, SK - 2: Before site prep and seedingdeployment                       Soil Impact – Salt (ECe ~ 10 dS/m)
Weyburn, SK - 2: PEPS deployment     – One month                    Soil Impact – Salt (ECe ~ 10 dS/m)
Weyburn, SK - 2: PEPS deployment       – 3 Months                     Average NaCl in leaf tissue = 23 g/kg               ...
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
Swathing/Mowing
Bailing
At salt sites, cut grass is removed from the site•    ~ 4000 lbs. of grass were removed from this site•    At remote sites...
Fall sampling and fall site work                            Sampling to get end                            of season sampl...
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
Sample Analysis                  • Analysis of PHC and                    salt impacted soils                  • Soil PHC ...
QA/QC Analysis PHC              PHC samples are              analyzed in at least              two laboratories           ...
QA/QC Analysis Salt               Salt samples are               analyzed in at least               two laboratories      ...
Interpretation of                          remediation results    Sample analysis    (GC and QA/QC)                       ...
Phytoremediation of PHC (A) Microbial aerobic PHC degradation – rhizosphere supported by plants (B) Possible microbial oxy...
After PEPS    Before PEPS                                                       Interpretation of              2000       ...
Full Scale PEPS Deployment at Typical PHC Sites                                                Average       %        Site...
For salt: NaCl in leaves – leaves removed                  from the site2 to 4 ha site - 500 kg of salt (NaCl) off the sit...
Full Scale PEPS Deployment at Typical Salt Sites                                                Average       %        Sit...
Why Use PEPS?• Scientifically proven and tested SOP• Verified methods for PHC and/or salt impacted sites.• Remediations at...
Thank youPlease visit us at the Earthmaster booth          for more information
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Greenberg chemistry of phc salt rem tech 2012 15 oct final

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Greenberg chemistry of phc salt rem tech 2012 15 oct final

  1. 1. The Chemistry and Biology BehindSuccessful Phytoremediation of Petroleum and Salt Impacted Soils Bruce Greenberg, X-D Huang, K Gerhardt, X-M Yu, S Liddycoat, X Lu, B McCallum, G MacNeill, M Hannaberg, P Mosley, J Gurska, N Knezevich, P Gerwing and B Chubb Partners: Cenovus; ConocoPhillips; Lone Pine Resources; Baytex; TransEuro Energy; Solaction; Shell; Talisman; PennWest; Questerre Beaver River; Imperial Oil; Seaway Energy Services; MWH; SNC Lavalin; NSERC
  2. 2. The Phytoremediation Process
  3. 3. Phytoremediation • Volatilization • Phytodegradation • Chelation/compartment in leaves Salt • Translocation: root symplast xylem • Chelation/compartment in roots • Plant uptake soilroot • Rhizosphere ProcessesRhizodegradation - PHC • Bioavailability particlewater
  4. 4. Advantages of Phytoremediation 1. Soil quality improved 2. Driven by solar energy - suitable to most regions and climates 3. Cost effective 4. Plants provide sufficient biomass for rapid remediation; promote high rhizosphere activity 5. Reasonable time frames - 2 to 3 years 6. Can be used effectively at remote sites 7. Greenhouse gas storage: 6 tonnes per ha per year 8. Effective for remediation of PHC and salt – relevant to the energy industry
  5. 5. Development, Proof and Full Scale Application ofPGPR Enhanced Phytoremediation Systems (PEPS) Over 13 years of research with full-scale field remediations at each stage of development and application 1. PHC: sites in AB, BC, QC, MB, NWT and ON (2004-12) 2. Salt: sites in SK, AB and NWT (2007-12) Performing full scale remediations for > 7 yrs PEPS currently successfully operating at > 30 sites > 10 sites completed
  6. 6. The key to phytoremediation success -Transfer of the science from the lab to the field1. Strategies for aggressive plant growth in impacted and poor quality soils at full scale sites – PEPS Deployment by highly trained scientists2. Monitoring the progress of phytoremediation at each site – Following the chemistry3. Continuous improvement of our phytoremediation systems through scientific research
  7. 7. WEBi-Earthmaster-UW Partnership• Synergistic expertise in contaminated site remediation• Developed commercial phytoremediation technologies (PEPS)• > 10 years of research, development and full scale field implementation• Field proven systems• Research to continually improve PEPS
  8. 8. PGPR Enhanced Phytoremediation Systems (PEPS)Aggressive plant growth strategies leads to remediation Physical soil treatment: Seed bed preparation Phytoremediation: Growth of plants with PGPR Monitoring and remediation assessment: Environmental chemistry to follow PEPS from start–to–finish • PGPR: Plant growth promoting rhizobacteria. • Prevent the synthesis of stress ethylene. • PGPR are applied to the grass seeds prior to sowing NOT Bioaugmentation • Effect depth of remediation ~ 0.5 m
  9. 9. "So, what have we beendoing for the last year?"
  10. 10. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  11. 11. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  12. 12. Interaction of a PGPR Containing ACCDeaminase with a Plant Seed or Root Amino Acids Plant growth promoting rhizobacteria (PGPR) Exudation Natural, non-pathogenic strains of PGPR Cell Elongation and Proliferation (usually Pseudomonads) IAA IAA Amino Acids SAM We have isolated PGPRs from ON, AB, SK ACC Synthase and the NWT ACC ACC ACC Deaminase ACC Oxidase PGPR are applied to seeds prior to planting Ammonia and -ketobutyrate Ethylene Stress Bacterium Response Plant Tissue
  13. 13. PGPR Isolation Plant Growth-Promoting Rhizobacteria (PGPR) Isolation • Have great PGPRs already – assume better strains in environment • New PGPR continually being isolated • Successful strains are assayed further by DNA sequencing to identify the bacteria. Only those in Biosafety Level 1 used. All non-GMOs • Isolated from site rhizosphere soils - environments we work in so they are adapted to impacted soils • These will be PGPR we will use in the future • New IRAP funding for this research
  14. 14. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  15. 15. PGPR Regeneration & Validation Regeneration of proven PGPR isolates for field use • Currently have > 10 strains of proven PGPR • Every year before use in PEPS – must confirm they are healthy and retain key biological activities for active plant growth promotion • Assay for ACC Deaminase • Assay for Auxin production
  16. 16. PGPR Validation – Assess plant growth enhancement• Before use in PEPS: plant growth assays to assure PGPRs perform properly• PGPRs now ready for use in PEPS
  17. 17. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  18. 18. Seed Treating & Shipping • Treat seeds with proven and regenerated PGPR for field deployment • Only proven grass and cereal species are used • Mechanical seed treater efficiently and evenly coats the seeds • Dried seed rapidly produced
  19. 19. PGPR Seed Treatment QA/QCAliquots of PGPR-treated seeds assayed for plant growth enhancementto assure successful PGPR application
  20. 20. Seed Treating & Shipping • Treated seeds shipped to sites after QA/QC
  21. 21. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  22. 22. Disking/Tilling to prepare seed bed
  23. 23. Application of appropriate fertilizer
  24. 24. Sample soil for beginning of season PHC and/or Salt levels
  25. 25. Sow PGPR-treated seed
  26. 26. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  27. 27. Edson, AB – Before site prep and seeding All previous steps assure sites that looked like this….. Soil Impact – PHC (Diesel Invert; 85% F3)
  28. 28. PEPS Deployment at Edson, AB…..Look like this Soil Impact – PHC (Diesel Invert; 85% F3)
  29. 29. Weyburn, SK - 2: Before site prep and seedingdeployment Soil Impact – Salt (ECe ~ 10 dS/m)
  30. 30. Weyburn, SK - 2: PEPS deployment – One month Soil Impact – Salt (ECe ~ 10 dS/m)
  31. 31. Weyburn, SK - 2: PEPS deployment – 3 Months Average NaCl in leaf tissue = 23 g/kg Soil Impact – Salt (ECe ~ 10 dS/m)
  32. 32. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  33. 33. Swathing/Mowing
  34. 34. Bailing
  35. 35. At salt sites, cut grass is removed from the site• ~ 4000 lbs. of grass were removed from this site• At remote sites - removal by helicopter• At PHC sites, grass does not need to be removed
  36. 36. Fall sampling and fall site work Sampling to get end of season samples At same sampling points as used at beginning of season.
  37. 37. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  38. 38. Sample Analysis • Analysis of PHC and salt impacted soils • Soil PHC – CCME GC method • Soil Salt – ECe, SAR, Na and Cl • Tissue Salt: Analysis of plant samples to assess plant uptake of salt
  39. 39. QA/QC Analysis PHC PHC samples are analyzed in at least two laboratories Data sets compared to assure data quality Only if correlations are strong are the data accepted
  40. 40. QA/QC Analysis Salt Salt samples are analyzed in at least two laboratories Data sets compared to assure data quality Only if correlations are strong are the data accepted
  41. 41. Interpretation of remediation results Sample analysis (GC and QA/QC) PGPR isolationFall sampling and PEPS PGPR regeneration fall site work & validation Seed preparation Plant growth and shipping and monitoring Site prep, sampling and seeding
  42. 42. Phytoremediation of PHC (A) Microbial aerobic PHC degradation – rhizosphere supported by plants (B) Possible microbial oxygenation pathway of PHC to form a fatty acid A B Cn PHC PHC-degrading microbes [O] ω-OxGenase oxidized PHC OH OH PHC [O] DEHYD O2 HH oil O O droplet [O] ω-OxGenase OH OH Cn fatty O O acid cell membrane -oxidation 1 µm (width C30 to C40) Acetate Rhizo. Cytosol
  43. 43. After PEPS Before PEPS Interpretation of 2000 remediation dataPHC (mg/kg) 1500 Example of a 1000 Completed Site 500 All 10 sampling points below criteria after 0 remediation Jun 07 Jun 08 Oct 08
  44. 44. Full Scale PEPS Deployment at Typical PHC Sites Average % Site Analysis Date Notes (mg/kg) RemediationCompleted Sites CCME F3 Spring 2007 1500 5 of 10 sample points above Tier 1 criteria Edson 33.33% CCME F3 Fall 2008 1000 All sample points met Tier 1 criteria CCME F3 Spring 2007 900 6 of 15 sample points above criteria Hinton 2 44.44% CCME F3 Fall 2008 500 All sample points met Tier 1 criteria EPH(C10-19) Spring 2009 6500 12 of 12 sample points above Tier 1 criteria Dawson 1 EPH(C10-19) Fall 2011 550 91.54% 1 of 12 sample points above Tier 1 criteria Examples of EPH(C19-32) Spring 2009 2500 11 of 12 sample points above Tier 1 criteria EPH(C19-32) Fall 2011 700 72.00% All sample points met Tier 1 criteria Completed Sites CCME F3 Spring 2007 900 4 of 11 sample points above Tier 1 criteria Peace River 78.89% CCME F3 Fall 2008 190 All sample points met Tier 1 criteria CCME F3 Spring 2009 550 3 of 3 sample points above criteria Quebec City 49.09% CCME F3 Fall 2009 280 All sample points met Tier 1 criteriaSites in Progress CCME F2 Spring 2010 1100 10 of 10 sample points above Tier 1 criteria 77.27% CCME F2 Fall 2010 250 6 of 10 sample points above Tier 1 criteria Hinton 1 CCME F3 Spring 2010 3200 9 of 10 sample points above Tier 1 criteria 56.25% CCME F3 Fall 2010 1400 3 of 10 sample points above Tier 1 criteria CCME F2 Spring 2009 1400 78.57% 8 of 8 sample points above Tier 1 criteria Examples of CCME F2 Fall 2010 300 4 of 8 sample points above Tier 1 criteria Swan Hills CCME F3 Spring 2009 2550 64.71% 7 of 8 sample points above Tier 1 criteria Sites in progress CCME F3 Fall 2010 900 1 of 8 sample points above Tier 1 criteria EPH(C10-19) Spring 2009 6500 15 of 15 sample points above Tier 1 criteria 46.15% EPH(C10-19) Fall 2011 3500 8 of 15 sample points above Tier 1 criteria Dawson 2 EPH(C19-32) Spring 2009 700 3 of 15 sample points above Tier 1 criteria 42.86% EPH(C19-32) Fall 2011 400 All sample point met Tier 1 criteria EPH(C10-19) Spring 2009 7000 11 of 12 sample points above Tier 1 criteria 81.43% EPH(C10-19) Fall 2011 1300 5 of 15 sample points above Tier 1 criteria Dawson 3 EPH(C19-32) Spring 2009 3500 12 of 12 sample points above Tier 1 criteria 57.14% EPH(C19-32) Fall 2011 1500 6 of 12 sample points above Tier 1 criteria EPH(C10-19) Spring 2010 1600 8 of 20 sample points above Tier 1 criteria 25.00% EPH(C10-19) Fall 2010 1200 6 of 20 sample points above Tier 1 criteria Beaver River EPH(C19-32) Spring 2010 850 8 of 20 sample points above Tier 1 criteria 35.29% EPH(C19-32) Fall 2010 550 3 of 20 sample points above Tier 1 criteria Average Remediation = 34 % per year
  45. 45. For salt: NaCl in leaves – leaves removed from the site2 to 4 ha site - 500 kg of salt (NaCl) off the site in the plantsper year
  46. 46. Full Scale PEPS Deployment at Typical Salt Sites Average % Site Analysis Date (dS/m) RemediationCompleted Sites ECe Spring 2008 7.7 Nota 70.13% ECe Fall 2010 2.3 Examples of ECe Spring 2009 14.5 Completed Sites Provost 44.83% ECe Fall 2009 8Sites in Progress ECe Fall 2010 13.5 Weyburn 1 22.22% ECe Fall 2011 10.5 ECe Fall 2010 6.9 Weyburn 2 14.49% ECe Fall 2011 5.9 Weyburn 3 ECe Fall 2010 13.5 10.37% Examples of ECe Fall 2011 12.1 ECe Fall 2010 14.3 Sites in progress Weyburn 4 11.89% ECe Fall 2011 12.6 ECe North, Sp 2010 5.2 13.46% ECe North, F 2011 4.5 Red Earth ECe South, Sp 2010 4.2 9.52% ECe South, F 2011 3.8 ECe Spring 2008 5.5 Kindersley 27.27% ECe Fall 2009 4 ECe Spring 2007 17.6 Cannigton Manor 32.95% ECe Fall 2008 11.8
  47. 47. Why Use PEPS?• Scientifically proven and tested SOP• Verified methods for PHC and/or salt impacted sites.• Remediations at all sites have been successful; > 30 sites.• Phytoremediation costs < half the cost of landfilling.• Liability is reduced, not transferred to a landfill.• Cost effective at remote sites.• Enhanced CCME BOC method – phytoremediation will meet Tier 1 criteria.• Tier 2 approach – will work – After PEPS brings F3 levels ≤ 2500 mg/kg no plant toxicity.
  48. 48. Thank youPlease visit us at the Earthmaster booth for more information
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