Transcript File - Amazon Mycorenewal Project

BIOREMEDIATION OF PETROLEUM CONTAMINATED SOIL
AND WATER
Robert Rawson,
President of Bioremediation International
[email protected]
Alexis Gropper,
[email protected]
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Bob Rawson
• President: International Wastewater Solutions Corporation, and BioRemediation International.
• Director of Bio-Remediation Research for IOS-Corporation in
Collaboration with CINVESTEV, and PRG Environmental Engineering,
Mexico.
• Inventor of US Patent No. 7,658,851 “AN APPARATUS AND METHOD
FOR GROWING BACTERIA”, for use in wastewater treatment,
biological pest management, bioremediation of soil and wastes.
• Inventor of US Patent No. 8114659 “A DEVICE AND METHOD FOR
THE CATALYTIC TREATMENT OF A MEDIA”.
• Vice President: Northern California River Watch, Environmental
Expert Witness for Clean Water Act, ESA, RCRA litigation.
• Environmental Technology Instructor at Santa Rosa Junior College 27
years
• Grade V Wastewater Treatment Operator 36 years Experience.
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I wish to thank the Geographic Institute
of Universidad San Francisco de Quito
alongside El Frente, UDAPT, and
everybody involved with organizing and
participating in this program,
“Reconstrucción Socio-Ecológica en el
Norte de la Amazonía Ecuatoriana”.
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Review
• Advantages of Bioremediation
• International Success of IOS-500 Bacteria
• Growing Methods and Applications of IOS-500
Bacteria
• Non-pathogenic certifications of IOS-500 Bacteria
• Multi-Kingdom Approach to Site Specific
Treatments
• Potential Proposals
Advantages of Bioremediation to Address
Multiple Problems Efficiently
Remediation efforts can serve multiple benefits:
• Compensate the “Affected People” for the enormous health
consequences that have been inflicted on them through:
Healing environment, Providing Jobs, Economic Stimulation
Optimizing the Labor and Capital needed to remediate the
contaminated environment in favor of the affected communities,
instead of strategies of highest economic gain distributed to few
people.
Production Possibilities Curve to clean up the Petroleum contaminated
The Y Axis of this
Graph represents the
money spent on
Equipment, Chemicals
and Energy required to
clean up a given
quantity of
contaminated soil.
OUTSIDE CAPITAL
Greater
Benefits
Expanding out from
Curves Q0 to QL show
greater Benefits like
the Amount of Soil
Bio-Remediated for a
specific combination
of Money as Capital
and Labor.
The X axis represents money paid to workers in the form of
wages and benefits to clean up a given unit of contaminated
soil.
WAGES and LOCAL EMPLOYMENT
• BOBS SCHEME
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Disadvantages of Conventional
Remediation Methods
• high chemical, mechanical and transportation
costs
• offsite and/or onsite pollution
• not true remediation (burning, chemical,
burying, relocating)
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International Success of IOS-500
Bacteria on Petrol-Contaminated Soils
• We will demonstrate several systems for
growing and implementing non-toxic bacterial
methods that are adequate for petroleum
spills on land and in various aquatic
environments.
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Santa Alejandra Swamp
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GROWING METHODS: Composting as
Bacterial Farming
• Utilizing bacteria in a composting process to
treat waste products such as petroleum
contaminated soil.
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BP Cleanup Site in Egypt.
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DILUTION!
Bacteria and Carbon are the magic ingredients.
It is necessary to dilute the concentration of
contaminants below a level of 15% and provide Carbon
mass to support bacterial survival and augmentation
(Cow and Chicken manure are excellent supplements)
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The Fire Triangle Analogy
Critical components for the survival of aerobic Bacteria
(like us we need food and oxygen to survive)
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Why Bioremediation is an Art
Composting microorganisms require the correct mix of carbon, nitrogen, oxygen and water
to operate. The recipe:
•
•
•
•
•
Blending the correct ratios of raw materials during windrow formation is one of the key
steps to successful composting.
According to literature, the optimum recipe will have a carbon-to-nitrogen (C:N) ratio of
between 20:1 and 30:1. This means there will be between 20–30 units of carbon, by mass,
for every unit of nitrogen present.
The optimum moisture content of the recipe is 40-60%. Feel is the best measure for
determining proper moisture, because clay and sand hold different amounts of water and
feel different in the hands.
The materials must be blended or piled to allow for oxygen to infiltrate the pile —
materials should not be compacted in a compost pile.
Observations of poor practices by local contractors practicing in Lago Agrio
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1) Dilution with Carbon sources and
application of IOS-500 Bacteria
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Example of Windrow layout and
turning.
2) Aeration
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3) Taking the Temperature of the Pile
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BP Cleanup Site in Egypt.
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Windrow Composting Rows
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WHAT ARE THE IOS-500 BACTERIA????
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IOS-500 TM contains
• Pseudomonas species Breaks down chemical
ring structures like phenol and nitrogenous
compounds. Capable of plasmid transfer.
(sharing information with other bacteria and
borrowing from them)
• Bacillus Species- Breaks down fats, oils,
greases, and cellulose.
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Aromatic Ring Compounds
Benzene
Naphthalene
Anthracene
Pyridine
Quinoline
Isoquinoline
Pyrazine
Quinoxaline
Acridine
Pyrimidine
Quinazoline
Pseudomonas in the aerobic root zone
of soil
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Bacillus Subtilus Enzymes for bio-remediating lipids.
2.4.
Metabolism of lipids BG10013 yybR; unknown; similar to ester hydrolase [SP:P37486] BG10305 bkdB, bfmB2, bfmBB, bkd; branched-chain alpha-keto acid
dehydrogenase E2 subunit (lipoamide acyltransferase) [EC:2.3.1.-] [SP:P37942] BG10306 bkdAB, bfmB1b, bfmBAB, bkd; branched-chain alpha-keto acid dehydrogenase E1 subunit (2oxoisovalerate dehydrogenase beta subunit) [EC:1.2.4.4] [SP:P37941] BG10307 bkdAA, bfmB1a, bfmBAA, bkd; branched-chain alpha-keto acid dehydrogenase E1 subunit (2-oxoisovalerate
dehydrogenase alpha subunit) [EC:1.2.4.4] [SP:P37940] BG10632 ywfH, ipa-86r; unknown; similar to 3-oxoacyl- acyl-carrier protein reductase [SP:P39644] BG10646 glpQ, ybeD;
glycerophosphoryl diester phosphodiesterase [EC:3.1.4.46] [SP:P37965] BG10679 lip, lipA; extracellular lipase [EC:3.1.1.3] [SP:P37957] BG11012 pssA, pss; phosphatidylserine synthase
[EC:2.7.8.8] [SP:P39823] BG11013 psd; phosphatidylserine decarboxylase [EC:4.1.1.65] [SP:P39822] BG11040 dgkA, yqxF; diacylglycerol kinase [EC:2.7.1.107] [SP:P19638] BG11048 yoxD;
unknown; similar to 3-oxoacyl- acyl-carrier protein reductase [SP:P14802] BG11153 scoA, yxjD; probable succinyl CoA:3-oxoacid CoA-transferase (subunit A) [EC:2.8.3.5] [SP:P42315] BG11154
scoB, yxjE; probable succinyl CoA:3-oxoacid CoA-transferase (subunit B) [EC:2.8.3.5] [SP:P42316] BG11225 ycsD; unknown; similar to hydroxymyristoyl-(acyl carrier protein) dehydratase
[SP:P42961] BG11239 acdA; acyl-CoA dehydrogenase [EC:1.3.99.-] [SP:P45867] BG11305 ywiE; unknown; similar to cardiolipin synthetase [SP:P45860] BG11310 ywjE; unknown; similar to
cardiolipin synthetase [SP:P45865] BG11319 mmgA, yqiL; acetyl-CoA acetyltransferase [EC:2.3.1.9] [SP:P45855] BG11320 mmgB, yqiM; 3-hydroxybutyryl-CoA dehydrogenase [EC:1.1.1.157]
[SP:P45856] BG11321 mmgC, yqiN; acyl-CoA dehydrogenase [EC:1.3.99.-] [SP:P45857] BG11373 pgsA, ymfN; phosphatidylglycerophosphate synthase [EC:2.7.8.5] [SP:P46322] BG11383 accB,
fabE, yqhW; acetyl-CoA carboxylase (biotin carboxyl carrier subunit) [EC:6.4.1.2] [SP:P49786] BG11384 accC, yqhX; acetyl-CoA carboxylase (biotin carboxylase subunit) [EC:6.4.1.2]
[SP:P49787] BG11417 ykhA; unknown; similar to acyl-CoA hydrolase [SP:P49851] BG11535 fabG, ylpF; beta-ketoacyl-acyl carrier protein reductase [EC:1.1.1.100] [SP:P51831] BG11536 acpA,
acpP; acyl carrier protein [SP:P80643] BG11611 ugtP, ypfP; UDP-glucose diacylglycerol glucosyltransferase [SP:P54166] BG11701 yqhM; unknown; similar to lipoate protein ligase [SP:P54511]
BG11714 yqiD; unknown; similar to geranyltranstransferase [SP:P54383] BG11719 yqiK; unknown; similar to glycerophosphodiester phosphodiesterase [SP:P54527] BG11722 ptb, bkd, yqiS;
probable phosphate butyryltransferase [EC:2.3.1.19] [SP:P54530] BG11723 bcd, bkd, yqiT; leucine dehydrogenase [SP:P54531] BG11724 buk, bkd, yqiU; probable branched-chain fatty-acid
kinase (butyrate kinase) [SP:P54532] BG11725 lpdV, bkd, yqiV; probable branched-chain alpha-keto acid dehydrogenase E3 subunit (dihydrolipoamide dehydrogenase) [SP:P54533] BG11746
yqjQ; unknown; similar to ketoacyl reductase [SP:P54554] BG11836 fabD, ylpE; malonyl CoA-acyl carrier protein transacylase [EC:2.3.1.39] [SP:P71019] BG11843 plsX, ylpD; involved in fatty
acid/phospholipid synthesis [SP:P71018] BG11946 lcfA; long chain acyl-CoA synthetase BG11951 lipB, yfiP; extracellular esterase [EC:3.1.1.1] BG12023 yclB; unknown; similar to phenylacrylic
acid decarboxylase BG12080 ydbM; unknown; similar to butyryl-CoA dehydrogenase BG12089 acpS, ydcB; probable holo-acyl carrier protein synthase [SP:P96618] BG12143 ydeP; unknown;
similar to cinnamoyl ester hydrolase BG12221 fabL, yfhR, ygaA; enoyl-acyl carrier protein reductase [EC:1.3.1.9] BG12241 yisP, yucD; unknown; similar to phytoene synthase BG12331 ysiB;
unknown; similar to 3-hydroxbutyryl-CoA dehydratase BG12456 ywhB; unknown; similar to 4-oxalocrotonate tautomerase BG12483 ywnE; unknown; similar to cardiolipin synthase BG12496
ywpB; unknown; similar to hydroxymyristoyl-(acyl carrier protein) dehydratase BG12557 accA; acetyl-CoA carboxylase (alpha subunit) BG12575 cdsA; phosphatidate cytidylyltransferase
[EC:2.7.7.41] BG12679 sqhC; squalene-hopene cyclase [EC:5.4.99.-] BG12729 cypC, ybdT; fatty acid beta-hydroxylating cytochrome P450 BG12809 ydzF; unknown; similar to cinnamoyl ester
hydrolase BG12914 yfjR; unknown; similar to 3-hydroxyisobutyrate dehydrogenase BG12994 yhaR; unknown; similar to enoyl CoA hydratase BG13021 yhdO; unknown; similar to 1acylglycerol-3-phosphate O- acyltransferase BG13029 yhdW; unknown; similar to glycerophosphodiester phosphodiesterase BG13048 fabHB, yhfB; beta-ketoacyl-acyl carrier protein
synthase III [EC:2.3.1.41] BG13055 yhfJ; unknown; similar to lipoate-protein ligase BG13057 yhfL; unknown; similar to long-chain fatty-acid-CoA ligase BG13063 yhfS; unknown; similar to
acetyl-CoA C-acetyltransferase BG13064 yhfT; unknown; similar to long-chain fatty-acid-CoA ligase BG13127 fabHA, yjaX; beta-ketoacyl-acyl carrier protein synthase III [EC:2.3.1.41]
[SP:O34746] BG13128 fabF, yjaY; beta-ketoacyl-acyl carrier protein synthase II BG13152 fabI, yjbW; enoyl-acyl carrier protein reductase [EC:1.3.1.9] [SP:P54616] BG13173 yjdA, yidA;
unknown; similar to 3-oxoacyl-acyl-carrier protein reductase BG13328 ykwC; unknown; similar to 3-hydroxyisobutyrate dehydrogenase BG13408 uppS, yluA; probable undecaprenyl
pyrophosphate synthetase [EC:2.5.1.31] [SP:O31751] BG13429 ymfI; unknown; similar to 3-oxoacyl- acyl-carrier protein reductase BG13457 yngF; unknown; similar to 3-hydroxbutyryl-CoA
dehydratase BG13458 yngG; unknown; similar to hydroxymethylglutaryl-CoA lyase BG13460 yngI; unknown; similar to long-chain acyl-CoA synthetase BG13461 yngJ; unknown; similar to
butyryl-CoA dehydrogenase BG13518 des, yocE; membrane phospholipid desaturase BG13523 yocJ; unknown; similar to acyl-carrier protein phosphodiesterase BG13544 yodR; unknown;
similar to butyrate-acetoacetate CoA-transferase BG13545 yodS; unknown; similar to 3-oxoadipate CoA-transferase BG13870 ytkK; unknown; similar to 3-oxoacyl- acyl-carrier protein
reductase BG13900 ytpA; unknown; similar to lysophospholipase BG13926 accD, yttI; acetyl-CoA carboxylase (beta subunit) BG14022 yusJ; unknown; similar to butyryl-CoA dehydrogenase
BG14023 yusK; unknown; similar to acetyl-CoA C-acyltransferase BG14024 yusL; unknown; similar to 3-hydroxyacyl-CoA dehydrogenase BG14029 yusQ; unknown; similar to acyloate
catabolism BG14030 yusR; unknown; similar to 3-oxoacyl- acyl-carrier protein reductase BG14031 yusS; unknown; similar to 3-oxoacyl- acyl-carrier protein reductase BG14063 yvaG;
unknown; similar to 3-oxoacyl- acyl-carrier protein reductase BG14144 yvrD; unknown; similar to ketoacyl-carrier protein reductase
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Growing and Application Methods of
the IOS-500
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IOS-500 microbes are grown with a White Knight inoculator contained inside
a Nursery Tank such as pictured. A microbial inoculation and aeration device
is depicted in this picture.
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Multi-Kingdom Applications to Site
Specific Treatments
• Bacteria (bio-remediation)
• Mushrooms (myco-remediation)
• Plants (phyto-remediation)
Ecosystem pioneers and their roles in
decomposition and distribution of the nutrient
cycle
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Possibilities with
Mycoremediation
Chemical structure
of lignin in wood
Chemical structures
of petroleum
Documenting species thriving directly
in oil spills
Taking it from Laboratory (ex-situ) to
In-situ
• In the laboratory
• Controlled Experiments(Ex-Situ)
• At a contaminated site (In-Situ)
Myco-Reactor Array
This conference has provided a unique
collaboration between local, international and
multi-disciplinary experts >> a conference of
complimentary strategies
We’ve seen so many examples of projects
terminating prematurely due to various reasons,
perhaps due to temporary international help
leaving, or funds drying out…
How do we guarantee the continuation of these
social and environmental projects? Short term,
Long term planning.
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Potential proposals
• Present here today we have universities, indigenous, regional organization
leaders, municipalities-- each with their individual strengths.
• A Communication platform to elaborate on the social and environmental
problems to work to find the adequate solutions
• Ideally, an institution where we can collectively train local leaders,
organizations, students, professors in these multi-disciplinary themes and
investigations
• Ideally, a location for on-site remediation demonstrations and continued
training
Criteria: security, electricity, containment, capacity, credibility, etc.
(potential sites like Municipality of Cascales where criteria are met)
• Such pilot projects can provide important opportunity to organize our
collective strategies in preparation for the larger scale remediation that
will be possible when the Texaco Case is successfully concluded.
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A Proposed Example of Stacking Multiple Functions that
hypothetically could provide jobs for Affected Communities, and
improve the economy.
• Grow Enormous numbers of Bacteria using waste products in the manner
that Alvaro Borja is in process of demonstrating at the City of Cascales and
then using these bacteria to treat the landfill leachate to reduce the
strength of the pollution being discharged to rivers.
• These same bacteria can then be used in many other applications
including: Bio-Remediation of Petroleum contaminated soil, and in
agriculture to treat fungal and bacterial plant pathogens including Black
Pod which is attacking the Cacao Crops, and other agricultural
commodities.
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• WE APPRECIATE YOUR PRESENCE, YOUR
ATTENTION, AND YOUR FUTURE
COLLABORATION OF IDEAS AND PROJECTS.
Robert Rawson,
President of Bioremediation International
[email protected]
Alexis Gropper,
[email protected]
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Controlling temperature is important
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Windrow pile construction.
• 4 feet (1.25 meters) or more high, by as much
as 12 feet (3.5 meters) across in layers of
matrix such as manure or green chop and
contaminated soil. I like a 9 foot high pile.
• Consider Mass Effect in determining the size.
More mass more heat and more moisture
retention. Cold and warm climate are
considerations.
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