This project is supported by the European Union under the Food, Agriculture and Fisheries and Biotechnology theme of the 7th Framework
Programme for Research and Technological Development under Grant Agreement no. 312139
Integrated biotechnological solutions for combating marine oil spills
Prof. Nicolas Kalogerakis, Coordinator School of Environmental Engineering Technical University of Crete
3rd Conference on "Ocean of Tomorrow" Projects, 11 November 2015 , Brussels
Weathering processes & period of activation The essence of
KILL•SPILL
To turn bioremediation from a weathering process into an attractive technology, several “problems” must be overcome (by human intervention):
Speed up the growth of indigenous hydrocarbon degraders or add (previously grown & lyophilized degraders) - bioaugmentation.
Add missing nutrients (primarily N&P) in oligotrophic environments - biostimulation.
Disperse oil into fine droplets (surfactants or biosurfactants)
Oxygen limitations can be a real challenge.
Can we make bioremediation a Green Technology?
Kill•Spill Project Focus / Objectives
1. Review State-of-the-Art & identify technology gaps
2. Develop techniques (biosensors & chemical) to monitor hydrocarbon degraders & hydrocarbon degradation
3. Develop novel bio-based dispersants
4. Develop novel bioremediation agents
5. Develop solutions for sediments
6. Develop multi-functional bioremediation agents
7. Test their toxicity
8. “Mandatory” Field Testing of Technologies Emergency Response Companies as Kill•Spill partners: EPE (Greece) & MMB (Norway)
Development of biosensors for the in-situ monitoring of hydrocarbons at contaminated sites
KIL
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Construction of plasmids encoding sensor/promoter::gfp pairs that, when introduced into a suitable host, respond to the presence of hydrocarbons by producing Green Fluorescent protein.
Sevilla, E., Yuste, L., and Rojo, F., 2015; Marine hydrocarbonoclastic bacteria as whole-cell biosensors for n-alkanes; Microbial Biotechnology, 8: 693-706
Gas CO2
Biodegradation is associated with an enrichment of 13C in the remaining contaminant pool increasing 13C
Compound Specific Isotope Analysis
First KillSpill FISH kits are now
commercially available …
FISH protocols adapted to easy and standardized VIT®
format allows fast and specific on-site monitoring
Identification and quantification of various hydrocarbon-
degrading bacteria using developed test kits:
o Alcanivorax borkumensis
o Cycloclasticus pugetii
o Marinobacter sp.
o Oleispira antarctica
o Oleiphilus messinensis
o Thalassolituus oleivorans
KIL
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kit
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Green: Alcanivorax borkumensis, Yellow: Oleispira antarctica; Total magnification 1000 x
Green: Alcanivorax borkumensis, Yellow: Oleispira antarctica; Total magnification 1000 x
Microbial Biosurfactants: Rhamnolipids
Sophorolipids
Biosurfactants + Oxygel (O2 delivery) + treated CaCO3
(adsorption/sinking agent) + hydrocarbon degrading bacteria.
Several plant surface active extracts/oils from BioBased Europe Ltd (a Scottish company).
Surface active products to date K
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Strain Biosurfactants (main congener) Yield (g/L)
Pseudomonas (aeruginosa PAO1) Rhamnolipids (Rha-Rha-C10-C10) 2-3
Burkholderia (Thailandensis E264) Rhamnolipids (Rha-Rha-C14-C14) 2-4
Candida bombicola Sophorolipids 20-100
Enhancing aerobic biodegradation with Oxygen releasing compounds: Gel form
Oxygel™
With additives (solvents & surfactants
from Bio-Based):
Suitable for free oil absorption into its matrix
& Oxygen release for enhanced degradation.
Without additives:
Suitable for injection into
sediments
Oil desorption & degradation
stimulation by oxygen release
Slow O2 release in H2O
Slow-release microparticles for the slow release of HC-degrading cells, biosurfactants, fertilizers, enzymes and e-acceptors
Polycaprolactone (PCL) COREs
PCL
Alginate HC-deg MOs
Nutrients (SRFs)
Kill•Spill Snorkel Proof of concept: a novel bio-electrochemical approach to
accelerate in situ the degradation of petroleum hydrocarbons in anoxic marine sediments
Large scale application considerations: Radius of influence & geometry
Development of a “smart gate” system allowing the release of the nutrient in the contact of an organic phase
P
P
PP
P
P
P
P
P
P
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PGATEOPENING
AQEUOUSPHASE ORGANICPHASE=
SELF-ASSEMBLY
SILICAPOLYCONDENSATION
CALCINATION
C16H33
N+
Biostimulation of crude oil contaminated beach sand
Biostimulation with N & P Biostimulation with N & P + rhamnolipids
Kill•Spill Sediment Cleaner OXIC (inside)
ANOXIC (outside)
Project co-funding by the European Union within the 7th Framework Programme under Grant Agreement No. 312139 is gratefully acknowledged
www.killspill.eu
Kill•Spill - Integrated Biotechnological Solutions for Combating Marine Oil Spills
Thank you for your attention!
All 28 Kill•Spill products developed or currently under development can be viewed at
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