1. Bioremediation: The current and the future Graeme Paton Technical Director, Remedios Diarmuid O’Sullivan Senior Environmental Consultant

2. Bioremediation “ The act of stimulating the metabolism or co-metabolism of contaminants in the natural environment. Can be ex-situ, in-situ or involve natural attenuation” 6 months - 3 years (ex-situ usually quicker)

3. Decomposition of biopolymers Cell Oxidation Endoenzyme Exoenzyme Dimers Monomers Endoenzyme Exoenzyme Extracellular enzymes ?? ½ O 2 H2OH2O CO 2

4. Tool Kit

5. Induction Catabolic biosensors enable a prediction of the bioavailability of the target molecule in that matrix Hence if the genes for degradation are in the matrix, degradation is very likely

6. Tool Kit

7. Procedure for lux-based assay % of the control luminescence 100 % 10 % Increasing toxicity

8. Tool Kit

9. Paton et al 2003 MPN- Tetrazolium

10. Site Location- Oil spill 1

11. ToxicityToxicity mpnmpn InductionInduction TPHTPH Tool Kit- at oil spill

12. Kuwait Oil Lake- Oil spill 2 During the first Gulf war (1990-1991) 50 million tonnes of soil was contaminated Air-borne oily material contaminated one thousand square kilometres of soil.

13. Systematic Grid

14. Can we predict? Bioremediation (f) [TPH] (f) availability (f) degraders (f) constraints BF= Induction x [TPH] mpn x (1-EC)

15. Prediction Once all of the data have been integrated an estimation of the likely case for degradation can be made Iroegbu et al., 2003

16. TPH Week 0Week 15

17. Prediction v Actual

18. Prediction There is a close relationship between the predicted value and the actual hydrocarbon concentration. This could define the end-point Regular monitoring confirmed the progress of this work This has been extended to over twenty trials to assess the value of the data

19. Max specific deg rate mg TPH/ day Bioremediation Factor Prediction

20. Max specific deg rate mg TPH/ day Bioremediation Factor Prediction

21. Gaswork Site- 10,000 tonnes

22. Bulk chemical- 2,500 tonnes

23. Coupling Technologies

24. The Big Issues On four occasions we have been required to seed/ augment a biological treatment process. This is a growing issue and a commercial opportunity.

25. Bioaugmentation Agents Inoc. Source Pre-condition 1, 2, 3, 4 Substrate Remedios Nutrients SoilSolution

26. Examples of Data

27. What does this mean? Oil Technics- cfu= 10 8 ; mpn- 10 7 Supplier 1- cfu= 10 6 ; mpn- 10 4 Supplier 2- cfu= 10 6 ; mpn- 10 5 Supplier 3- cfu= 10 4 ; mpn- 10 3 Oil Technics and precondition- cfu= 10 9 ; mpn- 10 7 Supplier 1 and precondition- cfu= 10 8 ; mpn- 10 5 Supplier 2 and precondition- cfu= 10 7 ; mpn- 10 6 Supplier 3 and precondition- cfu= 10 5 ; mpn- 10 3

28. Respiration Days

29. Degradation TPH Days

30. Max specific deg rate mg TPH/ day Bioremediation Factor Prediction

31. Max specific deg rate mg TPH/ day Bioremediation Factor Prediction

32. What does this mean Augmentation is possible Steps are required to refine Immobilisation of wood is not ideal Cost issues and value of application need to be considered Need to focus and develop the skill- base