Comparing Microbial Profiles from Five Service Station Sites
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The document discusses microbial profiling at five service station sites to evaluate biodegradation potential of contaminants such as BTEX and MTBE, utilizing various sampling methods to analyze microbial populations. Key findings included the prevalence of phenol hydroxylase and naphthalene dioxygenase genes, indicating potential for aerobic biodegradation, while also highlighting variations in microbial populations and biomass across different sites. Additionally, case studies illustrate troubleshooting and optimization strategies for remediation based on microbial data and geochemical conditions.
Comparing Microbial Profiles from Five Service Station Sites
- 1. Comparing Microbial Profiles from Five Service Station Sites Battelle Bioremediation Symposium Wednesday – May 24, 2017
- 3. • 30 days of in-well time • Place at level other samples are collected BioTrap® Samplers
- 4. • Real time sample collection • Low budget • Comparable accuracy Bio-flo Filter® Sampling
- 9. • Phenol Hydroxylase (qPHE) - Catalyzes further oxidation of BTEX compounds, indicates the potential for aerobic BTEX biodegradation. • Naphthalene dioxygenase (qNAH) - Catalyzes aerobic biodegradation of naphthalene and other PAHs by incorporation of oxygen into the aromatic ring. • MTBE utilizing PM1 (qPM1) - Targets Methylibium petroleiphilum PM1, one of the few bacteria isolated that is capable of growth on MTBE, indicates potential for aerobic MTBE biodegradation. • Total Eubacteria (qEBAC) – total biomass estimate. Four CENSUS Targets
- 10. • Total biomass averaged E+06 cells/ml or cell/bead regardless of soil type • Little difference between BioTrap® and Bio-flo ® filter • PHE gene function most prevalent at four sites/PHE increases possible • NAH gene function prevalent at only one site/NAH increase possible • PM1 organism typically at E+02 at sites where it was present • PM 1 increases found • Unusual conditions can exist Observations from the 5 Sites
- 11. • Total biomass averaged E+07 cells/ml or cells/bead regardless of soil type • PHE gene function most prevalent at 11 sites • NAH gene function most prevalent at 4 sites • PM1 organism typically at E+02 at sites where it was present. • Unusual conditions can exist Observations Compared to 15 Additional Sites
- 12. Well I.D. Date pH ORP DO TPH-g Benzene MTBE TBA Methane Sulfate Manganese (Dissolved) Manganese (Total) Iron (Total) Iron (II) Ferrous (units) (millivolts) (mg/L) (µg/L) (µg/L) (µg/L) (µg/L) (µg/L) (mg/L) (mg/L) (mg/L) (mg/L) (mg/L) MW-1 3/16/2012 6.6 (208.4) 0.59 2,200 56 13 2,800 3530 110 2.77 3.00 9.95 0.24 8/1/2012 11.66 (57.8) 28.48 400 17 11 470 52.9 780 <0.00500 0.0777 0.866 <0.100 10/18/2012 9.9 48 32.39 240 7.4 16 49 81.8 680 <0.00500 0.0190 0.220 <0.100 12/18/2012 8.35 106.9 15.2 53 0.73 2.1 19 22.2 650 0.0198 0.0905 0.714 <0.100 2/18/2013 8.04 129.8 10.84 52 <0.50 <1.0 <10 4.05 600 0.0147 0.0852 0.314 <0.100 Optimal Response
- 13. • PHE baseline was E+04 PM1 baseline was E+02 • PHE increased to E+07 • NAH baseline was E+02 • NAH increased to E+05 • EBAC baseline was E+07 PM1 increased to E+06 • EBAC increased to E+09 Microbial Profile near MW-1
- 14. Well I.D. Date pH ORP DO TPH-g Benzene MTBE TBA Methane Sulfate Manganese (Dissolved) Manganese (Total) Iron (Total) Iron (II) Ferrous (units) (millivolts) (mg/L) (µg/L) (µg/L) (µg/L) (µg/L) (µg/L) (mg/L) (mg/L) (mg/L) (mg/L) (mg/L) B2-A 3/15/2012 6.75 224.7 3.68 68 <0.50 3.9 38 10.9 270 0.553 0.946 0.995 <0.10 8/17/2012 6.63 (25.8) 1.33 120 0.60 1.6 1300 408 260 2.62 2.76 5.16 <0.100 10/18/2012 7.03 (6.7) 1.68 230 1.6 <2.0 1700 1090 250 2.81 2.99 8.07 7.14 12/18/2012 7.18 68.6 1.17 270 <1.0 <2.0 1700 950 280 2.53 2.88 11.4 5.26 2/18/2013 7.21 47.2 0.92 230 <0.50 1.9 1400 754 270 2.40 2.40 7.43 5.63 What’s up here?
- 15. • PHE baseline was E+03 PM1 baseline was E+02 • PHE decreased to E+02 • NAH baseline was E+02 • NAH decreased to E+01 • EBAC baseline was E+06 PM1 decreased to E+00 • EBAC decreased to E+04 Microbial Profile near B2-A
- 16. • MTBE/TBA remediation • (20 µg/L/6000 µg/L) • Lesser concentrations of • benzene and TPHg • Full suite of analytical and • field parameters • No degradation observed • No apparent response in geochemical or field data Case Study – Inactive Service Station
- 18. • Methane and carbon dioxide showed dramatic fluctuation across the site. • Possibly a microbial issue? • Investigation undertaken to evaluate microbial population – Total bacteria – MTBE/TBA degrader – Methanotrophs – bacteria that consume methane as a source of energy – Methanogens – bacteria that produce methane while degrading organic matter under anaerobic conditions Case Study – The cause?
- 19. • CENSUS – upgradient, treatment area, downgradient Remediation Troubleshooting Data
- 20. • Evidence of Deeply Reduced Aquifer • Bio-Geochemically - methanogenic • Raise REDOX (climb the REDOX ladder) – Satisfy O2 debt • Used a quick release O2 material to – rapidly satisfy REDOX demands and – reduce non-target degrader populations – improve target degrader populations • Use ORC Advanced for long-term O2 release – Maintain beneficial REDOX condition – Provide support for biodegradation of target COC’s Case Study - Optimization
- 23. Microbial data establish a baseline. Microbial data allow tracking of progress. Microbial data can help troubleshoot a site. SUMMARY
- 24. Questions and Answers Jack Sheldon jack.sheldon@anteagroup.com Acknowledgement – Jeff Friedman, Antea Group-Long Beach