Robert Rawson President of Bioremediation International iwssonicnet Alexis Gropper lexgropgmailcom 1 Bob Rawson President International Wastewater Solutions Corporation and BioRemediation International ID: 920309
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Slide1
BIOREMEDIATION OF PETROLEUM CONTAMINATED SOIL AND WATER
Robert Rawson,President of Bioremediation Internationaliws@sonic.netAlexis Gropper,lexgrop@gmail.com
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Slide2Bob
RawsonPresident: International Wastewater Solutions Corporation, and Bio-Remediation 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 yearsGrade V Wastewater Treatment Operator 36 years Experience.
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Slide3s
3I 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”.
Slide4Review
Advantages of BioremediationInternational Success of IOS-500 BacteriaGrowing Methods and Applications of IOS-500 BacteriaNon-pathogenic certifications of IOS-500 BacteriaMulti-Kingdom Approach to Site Specific TreatmentsPotential Proposals
Slide5Advantages 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.
Slide6The 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 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
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.Production Possibilities Curve to clean up the Petroleum contaminated Greater Benefits
Slide7BOBS SCHEME
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Slide8Disadvantages of Conventional Remediation Methods
high chemical, mechanical and transportation costsoffsite and/or onsite pollutionnot true remediation (burning, chemical, burying, relocating)8
Slide9International 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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Slide10Slide11Santa Alejandra Swamp
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Slide1212
Slide1313
Slide1414
Slide15GROWING METHODS: Composting
as Bacterial FarmingUtilizing bacteria in a composting process to treat waste products such as petroleum contaminated soil.15
Slide16BP Cleanup Site in Egypt
.
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Slide17DILUTION!
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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Slide18The Fire Triangle Analogy
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Critical components for the survival of aerobic Bacteria
(like us we need food and oxygen to survive)
Slide19Why Bioremediation is an Art
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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
Slide2020
1) Dilution with Carbon sources and
application of IOS-500 Bacteria
Slide21Example of Windrow layout and turning.
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2) Aeration
Slide2222
Slide233) Taking
the Temperature of the Pile
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Slide24BP Cleanup Site in Egypt
.
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Slide2525
Windrow Composting Rows
Slide26WHAT ARE THE IOS-500 BACTERIA????
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Slide27Slide2828
Slide29IOS-500
TM containsPseudomonas 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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Slide30Aromatic Ring Compounds
BenzeneNaphthalene
Anthracene
Pyridine
QuinolineIsoquinolinePyrazineQuinoxaline
Acridine
Pyrimidine
Quinazoline
Slide31Pseudomonas in the aerobic root zone of soil
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Slide32Bacillus 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 (2-oxoisovalerate 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 1-acylglycerol-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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Slide33Growing and Application Methods of the IOS-500
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Slide34IOS-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.
Slide3535
Slide3636
Slide3737
Slide38Multi-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 cycle38
Slide39Chemical structure of
lignin
in wood
Chemical structures of petroleum
Possibilities with
Myco
remediation
Slide40Documenting species thriving directly in oil spills
Slide41Taking it from Laboratory (ex-situ) to In-situ
In the laboratoryControlled Experiments(Ex-Situ) At a contaminated site (In-Situ)
Slide42Myco-Reactor Array
Slide43This 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.43
Slide44Potential 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 solutionsIdeally, 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. 44
Slide45A 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.45
Slide46WE APPRECIATE YOUR PRESENCE, YOUR ATTENTION, AND YOUR FUTURE COLLABORATION OF IDEAS AND PROJECTS.
Robert Rawson,President of Bioremediation Internationaliws@sonic.netAlexis Gropper,lexgrop@gmail.com
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Slide47Controlling temperature is important
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Slide48Windrow 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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