Enzymatic Bioremediation of Petroleum Hydrocarbons

Transcription

1 Enzymatic Bioremediation of Petroleum Hydrocarbons Paul W Sammarco Louisiana Universities Marine Consortium (LUMCON) Chauvin, LA USA psammarco@lumconedu, (985) Elizabeth Montgomery Lawrence Anthony Earth Organization (LAEO) Glendale-Montrose, CA USA montgomerylizzy@gmailcom William A Adams Remote Energy Security Technologies Collaborative (RESTCo) Ottawa, ONT Canada beckettslanding@gmailcom (613) Diane Wagenbrenner Lawrence Anthony Earth Organization (LAEO) Glendale-Montrose, CA USA diane@theearthorganizationorg (858)

2 Characteristics of Enzymatic Biotreatment of Oil Spills Biodegrades crude oil rapidly into smaller components and finally into CO2 and H2O Non-toxicity - demonstrated on fw and marine organisms fish, plankton, and bacteria Non-toxic to humans Stimulates population growth in naturally occurring bacteria Works to completion within a period of 4-6 weeks mechanism of action protects shorelines, beaches, marine life and human health within hours

3 Four Types of Remediation for Oil Spills NCP Product Types Dispersants Nutrient Additives Microbiological Cultures Enzyme Activities - Example OSE-II

4 Mechanism of Enzyme Bioremediation Action Contains - Bio-surfactants - Enzymes - Nutrients

5 Testin g Tests performed by numerous intl govt agencies and independent labs Non-Toxic - Safe for marine and aquatic organisms at effective applications levels - No specific precautions need be taken - No vapors emitted; thus, no inhalation problems - Safe for humans (OSHA) - No special handling or protective equipment or clothing required

6 Efficacy

7 Concentration of Analytes (ppm) Degradation of 96 Analytes through Time Using a Bioenzymatic treatment (OSE-II) vs Nutrient Controls 12 Non-Nutrient Control Nutrient Treatment 8 Bio-enzymatic Treatment (OSE-II) Time (Days) 3

8 TPH Biodegradation Tests Using Enzymatic Treatment of Crude Oil Performed by Independent Labs (OSE-II)

9 (University of Yamanashi, Japan

10 Dielectric Insul Oil Concentration (ppm) Enzymatic Bioremediation (OSE-II) Dissipation of Spilled Dielectric Insulating Oil (by GC/MS) 28 Days 6 Control 4 Nutrient Addit 2 OSE-II Time (Days) (US Dept Interior, BOEM; and Dept Envtl Sci, LA State Univ)

11 Toxicity

12 Pimephales promelas (Fathead Minnow)

13 Mortality of Pimephales promelas 2 (Y = X + 311X ) 1 Percent Morality (Angular Transform) _ 5 _ 25 _ 5_ LC-5 _ 1 1, 1, OSE-II Concentration (mg/l, log transform) 1,

14 Enzymatic Bioremediation (OSE-II) Toxicity Pimephales promelas (Fathead Minnow) 2 Mortality (no inds, n=2) LC5 = 11,8 mg/l 5 1, 5, 2, 1, 3, Concentration (OSE-II) (mg/l, log [X+1] transform)

15 Enzymatic Bioremediation (OSE-II) Toxicity - Mortality Pimephales promelas (Fathead Minnow) Mortality (no inds, n = 2) LC5 = 5,85634 mg/l 25 1, 4, 16, 5 2, 8, Concentration (OSE-II) (mg/l, Log1 + 1 transform)

16 Ceriodaphnia dubia (Freshwater water flea planktonic) nidae/gceriodaphnia/ceriodaphnia_dubia/ceriodaphniadubia html

17 Enzymatic Bioremediation Toxicity - Ceriodaphnia dubia (planktonic crustacean) (Using OSE-II) Y = 373/(1+e(-(x-4281)/168)) Percent Mortality LC-5 5 2, 4, 8, 16, Concentration (mg/l) (Sqrt Transform)

18 Enzymatic Biotreatment (OSE-II) Toxicity - Mortality Ceriodaphnia dubia (fw water flea) Mortality (no inds, n = 2) Concentration of Application and Efficacy 5 LC5 > 16, mg/l , 5 4, 2, 16, 8, Concentration (mg/l, Log1 + 1 transform) (Huther & Assocs, 28)

19 Mysidopsis bahia (Estuarine/marine mysid shrimp)

20 OSE-II Enzymatic Bioremediation Toxicity - Mysidopsis bahia (mysid shrimp) 1, mg/l Percent Mortality _ LC-5 = 2,1 25 1, 1, 1, Time (hrs) EnviroSystems Division Resource Analysts, Inc

21 Artemia salina (Freshwater brine shrimp, Sea Monkeys )

22 Enzymatic Bioremediation Toxicity - Artemia salina (planktonic crustacean) (Using OSE-II and Water) Percent Mortality mg/l Time (hrs) 5

23 Enzyme Bioremediation (OSE-II) Toxicity Mortality in Artemia salina (freshwater plankton) 2 OSE-II Alone 1 LC5 = > 1 mg/l } 5 { 25 mg/l LC5 = 126 mg/l No 2 Fuel Oil 15 Mortality (no inds) OSE-II and No 2 Fuel Oil (1:1) 25 Mortality (no inds, n = 2) Mortality (no inds, n = 2) 2 1 mg/l 4, mg/l LC5 = 294 mg/l Time (hrs) 48 48

24 Additional Test Organisms Oncorhyncus mykiss (Pacific salmon) Menidia sp (Neotropical Silverside) Daphnia magna (fw plankton) Mysidopsis sp Photobacterium phosphorum Gasterosteus aculeatus(estuarine/marine shrimp) m-negative phosphorescent (fw three-spined-stickleback) bacterium)

25 12 Enzymatic B ioremediatio n (OSE-II) LC-5s for Two Corexit Products and OSE-II Enzymatic Bioremediation Product, on Five Test Organisms Corexit 95 Corexit 9527 * LC-5 (ppm) Menidia Gasterosteus Sp aculeatus (Neotropical (three-spined silverside stickleback fish) fish) Mysidopsis Dahnia Photobacterium sp magna phosphoreum (mysid (planktonic (gram-neg shrimp) fw crustacean) phosphorescent bacterium) Oncorhynchus mykiss (salmon) *Contains 2-BTE Test Species

26 Toxicity to Humans

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28 (OSHA)

29 Bacterial Enhancement

30 Enzymatic Bioremediation (OSE-II) Bacterial Enhancement OSE-II Bacterial Populations (MPN, Log1) 1,, Nutrient Addit 1,, Control 1, Time (Days) (Bio-Aquatic Testing, NELAC Certified)

31 Enzymatic Biotreatment (OSE-II) Bacterial Response 1x16 OSE-II 3 Bacterial Count (MPN) 75x1 Nutrient Addit 5x13 Control 3 25x1 1 2 Time (Days) (US Dept Interior, BOEM; and Dept Envtl Sci, LA State Univ) 3

32 Respirocity Test Indicator of Oxygen-Enhanced Bacteria Oxygen Uptake (mg/l) Days: 1 3 Product Product 2 Comparative Products 3 Bioenzymatic Treatment (OSE-II)

33 Enhancement of Bacterial Population Growth on Crude Oil after application of Enzymatic Bioremediation treatment (OSE-II) (OSE-II)

34 Cost Comparisons BP/Deepwater Horizon Spill Est cost of treatment in situ burning, mechanical recove chemical dispersants, damage claims, fines - Est spillage 2M gallons - Total est cost of remediation - $42B Est cost of clean-up = $21/gal oil Cost of alternate approach Enzymatic Bioremediation - Total est cost of remediation - $8M Est cost of clean-up = $4/gal oil (Savings 981%)

35 Countries using Enzymatic Bioremediation on Oil Spills with Governmental Approval Australia Bahrain Colombia Greece Iraq Iran Kenya Kuwait Mexico New Zealand Nigeria Oman Philippines Qatar Saudi Arabia South Korea Trinidad and Tobago United Arab Emirates United Kingdom United States (Military Army, Navy, Coast Guard Air Force, Marines)

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38 Acknowledgements The Lawrence Anthony Earth Organization (LAEO) - Ms Diane Wagenbrenner Ms Elizabeth Montgomery Ms Barbara Wiseman Mr Steven Pedigo, The OSE-II Corporation Louisiana Universities Marine Consortium (LUMCON) The 216 Clean Gulf Conference Ms Cassie Davie

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40 Dispersants Cleanup % Toxicit y Toxic Enzymatic Bioremediat Mechanical ion 2-8% ~ 1% Toxic Non-toxic (derived from (detoxifies (To marine spp oil) oil quickly) & humans, particularly w 2-butoxyethanol 2-BTE

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42 Tier II Efficacy Data Enymatic Bioremediation Treatment (OSE-II) Percent Reduction after 21 Days Ch rys en la e ro m Ph at i en cs an thr en es Flu or en e Pr ist an e C3 Ph To yta tal ne n-p ara ffin s C18 To ta Percent Reduction (%) 1 Analytes

43 OSE-II Toxicity Test - LC-5 (mg/ L) Menidia beryllina and M bahia (Silversides- fish) Species Menidia beryllina Menidia bahia * Static * * Static renewal Duration (hrs) * * * *

44 Enzymatic Biotreatment (OSE-II) Juvenile Mysid Shrimp Toxicity - Survivorship 1 1, mg/l Mortality (no inds) 8 6 _ 4 LC5-2,1 mg/l 1, mg/l { mg/l 1 mg/l 1 mg/l mg/l (Control) Time (hrs) (EPA/NETAC)

45 Mechanism of Action (cont) Neutralizes toxicity to soil and aquatic or marine organisms Enzymes separate oil molecules and enhance their metabolic breakdown

46 Additional Advantages of Enzymatic Bioremediation No specific precautions need be taken No vapors emitted; thus, no inhalation problems A one-time application is sufficient to achieve effectiveness