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BIOREMEDIATIONANDTHEUSE
OFBIOAUGMENTATIONPRODUCTS
SUMMARY:
Bioremediationtechnologyencompassesthecontrolled,practicaluseofmicroorganismsforthebreakdownofchemical
pollutants.Thesevarioustechnologiesrelyonthebiodegradationactivitiesofmicroorganisms.Theyfocusonenhancing
existing, but slow, biodegradation processes in nature, or technologies that bring chemicals into contact with micro
organisms in reactors to effect a rapid degradation. The popular use of the term bioremediation relates to insitu
treatment of contaminated sites by the optimization of conditions that otherwise limit the biological breakdown of
pollutants.

INTRODUCTION:
Thegoalofbioremediationistodegradeorganicpollutantssothatremainingconcentrationsareeitherundetectableor,
ifdetectable,belowthelimitsestablishedassafebyregulatoryagencies.
Bioremediation is used for the breakdown or transformation of chemicals in soils, groundwater, wastewater, sludge,
industrialwastesystems,andgases.Typicalcandidatesforbioremediationaresitesthatarecontaminatedwithoiland
oil products, gasoline and its constituents, polycyclic aromatic hydrocarbons (PAH), chlorinated aliphatics such as
tetrachloroethylene(TCE)andperchloroethylene(PCE),andchlorinatedaromatichydrocarbons.Suchsitesarecurrently
recognized as hazardous to health and the environment. Soil contaminated with pesticides may also be suitable for
bioremediation.
Natural, indigenous microorganisms have the capacity to detoxify a variety of compounds. Bioremediation is a cost
effectivetechnologythatcanrestorelargecontaminatedareastothepristineconditionsthatmayhaveprevailedprior
tothepollution.Itisaflexibletechnology,asavarietyofprotocolsmaybedesignedtoaccommodatethespecificneeds
ofeachseparatecase.
Thetechnologyhasalonghistory,andiscurrentlyacceptedasadependablesolutioninmanycaseswherechronicor
acute problems exist. The largest field demonstration of bioremediation ever undertaken was carried out after the
groundingoftheoiltankerExxonValdezinPrinceWilliamSound,Alaska,1989.Atotalof37000tonsofcrudeoilwas
releasedandalargeproportionofthisspreadontotheadjacentshorelines.Thesubsequenttreatmentandmonitoring
unambiguouslyshowedthatbioremediationenhancedtherate,andimprovedtheextentofbreakdownofthemarine
oil spill. The same has been verified for polluted soils throughout the United States. Bioremediation of soil is often
carried out by mixing the contaminated material with different additives that promote the degradation process. The
fertilizedmassesareallowedtoreactforweeksandmonths,andtheindigenousmicroflorawilldetoxifyanddegrade
manyofthemostcommonorganicpollutantstoacceptablelimits.

BIOLOGICALDEGRADATION:
Inrecentyearsmethodsofbiologicalremediationofoilcontaminatedsoilshavebecomemoreandmoresignificantdue
totheirecologicalsafety.Therearetwoprincipalapproachestothebiologicaldegradationofhydrocarbonsinwaterand
soil: (1) introduction of specifically selected associations of microorganisms, which degrade various classes of
hydrocarbon pollutants (bioaugmentation); (2) activation of indigenous oiloxidizing microflora by providing optimal
conditionsforitsgrowth(biostimulation).Themosteffectivearebiotechnologiesincludingbothapproachese.g.useof
biofertilizercontainingmicrobialculturesandinorganicnutrients.

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It should be noted, that in cooler climatic zones and their typical shorter warm periods (120125 days), natural
attenuationofoilcontaminatedsoilsisslow.Therefore,biotechnologiesthatinvolvestimulationofhydrocarbon
oxidationbynaturaloiloxidizingmicroorganismsisessentialtosuccess.Severalagrotechnicalmethods,namely,tilling
andloosening;wateringanddrainagesystems;additionoforganicmaterials(straw,compostetc)andmineralfertilizers
coulddecreasethecontaminationlevelupto3040%duetotheoxidationofeasilydegradablepetroleumcomponents
(Christofietal.,1998).However,highmolecularparaffins,aromaticandpolycycliccompounds,withconcentrationsin
crude oil from 20 to 65 %, are not degraded for years. The above compounds bind to the soil particles, producing
hydrophobicresidues,andbecomenonbioavailabletomicroorganisms.Toincreasethebioavailabilityofhydrocarbon
pollutants, surfaceactive substances (surfactants) are used which allow desorption and solubilization of petroleum
hydrocarbons and thus facilitate their assimilation by microbial cells (Deschenes et al., 1995; Volkering et al., 1995;
Thibaultetal.,1996).Manychemicalsupplierspromotesyntheticsurfactantstobeusedforpurposeandtheycanbe
highlytoxicandnonbiodegradable(Chinetal.,1996).Applicationofsyntheticsurfactantsleadstotheaccumulationof
ecologically harmful compounds in soil. Until the recent development of Synergy Services BioSolv 801 Biosurfactant,
organic biodegradable surfactants were considered notably difficult to formulate a technology that would breakdown
largemassesofoilintomicroscopicmicellularoildropletsforbiologicalassimilation.TheSynergyServicesBioSolv801
Biosurfactantwasdevelopedbyoilfieldchemicalengineerswithyearsofexperienceinthedispersantandremovalof
paraffinandasphaltenedepositsfromformations,wellbores,andproductiontubulars.Withthisknowledgebeingvital
to their solution, Synergy Services teamed with key Microbiologists and Geneticists, in order to fill the need for a
biodegradableorganicsurfactanttechnologythataugmentsthebreakdownandspeedsbioassimilationofcrudeoilsand
otherorganicwastes.
EffectiveandecologicallysafebiosurfactantsproducedbyRhodococcusbacteriaisolatedfromoilextractionareaswere
successfully developed by a team of scientists years ago and never marketed due to collaboration problems. Our
developmentteamdiscoveredthatthesebiosurfactantspossesssignificantadvantagesoversyntheticones,particularly
biodegradability,stableactivityunderextremeenvironment conditions,awiderangeoffunctionalcharacteristicsand
thepossibilitytobeproducedfromnontraditionalandrelativelyinexpensiverawmaterials.Laboratorystudydataon
recovery of heavy oils (density 0.93 to 0.97 g/cm3) from oilcontaminated sand using Rhodococcus biosurfactant
complexesshowedthattherateofoilremovalexceedsthecontroldata1520fold(Ivshinaetal.,1998).
TwoRhodococcusstrainsisolatedfromaheavyoilextractingareawereusedforbiofertilizerproduction;R.erythrolopis
wasisolatedfromthebottomofcrudeoilwastestoragepitandtheR.ruberwasisolatedfromanoilcontaminatedsoil.
Theabilityofstrainstousepetroleumhydrocarbonsassolecarbonsourcewastestedonmineralagarcontaining0.5%
of individual compound (naphthalene was tested in a vapor phase). The growth of strains on crude oil was tested in
shake flask experiments (Mueller et al., 1992) and was shown to breakdown the heavy oils quickly with the added
benefitofthenaturallyoccurringbiosurfactantpresent.
SYNERGYSERVICESBIOREMEDIATIONTECHNOLOGIES:
Synergy Services is a manufacturer of pipeline cleaning, oilfield treatment, industrial water technologies, and
environmental treatment products. Our product offerings for the cleanup of petroleumbased contaminants in soil,
ground water and marine environments include BioSolv 801 Biosurfactant, BioSolv 808 Bioaugmentation Microbes,
and BioSolv 817 Fertilizer/Stimulant. These propriety formulations consist of enzymes, organic nutrients,
microorganisms, and stimulants that enhance the bioremediation processes. The technology of bioremediation is
confirmedscientificallysound bythe USEPAandnumerousenvironmentalagenciesaroundtheworld, consideredthe
mostenvironmentallybenignapproachtocleanup,andofferthegreatestlevelofcompetivenessinusecosteconomics.

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ENHANCEDBIOREMEDIATION
Synergy Services BioSolv 801 Biosurfactant is a complex derived from Rhodococcus (lipophyllic bacterium) cultures,
organic surfactants, dispersants, microbiological enzymes, and proprietary biostimulation additives. The Product is
designed to disperse and augment remediation of crude oil spills on water, soil, or solid substrate surfaces.
Biosurfactants disperse hydrocarbon contamination into micellular particles which are easily assimilated and
biodegraded by the indigenous microbes provided by nature or augmented by application of hydrocarbon degrading
microorganisms.BioSolv801isamultifacetedbioremediationenhancingagentwhichishighlyeffective,simpletouse,
costeffective and completely nontoxic. The product contains no bacteria or genetically engineered microbes, but
ratheritstimulatesindigenousbacteriatodegradepetroleumproducts.Theultimatemeasureofsuccesshasbeenthe
rapid disappearance of both target and nontarget contaminants without the buildup of toxic byproducts following
BioSolv801enhancedbioremediation.
BioSolv 801 Biosurfactant technology, while agreeably simple, is scientifically sound and highly costeffective. It is
environmentallyfriendly,nontoxic,andallitsingredientsareorganicinoriginandcompletelybiodegradable.
MECHANISMOFACTION
Soil and its complex microbial community can accept and degrade to some degree a variety of petroleumbased
pollutantsintonontoxicforms.Thereexistsinnaturemicrobialforms(i.e.,bacteria,yeast,molds,etc.)whichhavethe
capability to biodegrade many petroleumbased agents which are noxious to the environment. There are particular
families of microbes that Synergy Services markets under the trade name, BioSolv 808 Bioaugmentation Microbes
(consists of a proprietary blend of microbes known as Pseudomas, Arthobacter, Alcaligenes, Corynbacterium,
Flavobacterium,Achrombacter,Acinetobacter,Micrococcus,Nocardia,andMycobacterium)thatpossesstheabilityto
produce extracellular enzymes which leads to the breakdown (mineralization) of the petroleum compounds and
transforms them into food sources for the microbes. Upon biodigestion, all that is left of the waste is carbon dioxide
(CO2),water(H20),andmicrobialbiomass(fattyacidswhichareusedforafoodsourcebyothermicroorganisms).
Thebiodegradationofpetroleumbasedcompoundscanbeaccomplishedbothinoxygenrich(aerobic)andinoxygen
poor(anaerobic)environments.Theaerobicmineralizationofcontaminantsismuchfasterandmorecompletethanthe
anaerobicprocess.However,bothaerobicandanaerobictreatmentsmaybenecessaryinsomecases.
At sites contaminated with petroleumbased compounds, a small percentage of the overall bacterial population is
capableofutilizingthepetroleumcompoundsasafoodsourceforenergyandgrowth.Thislimitedbacterialpopulation,
combinedwithalackofenhancedstimulation,iswhatlimitsmostnormalbioremediationprojects.Withtheadditionof
BioSolv801BiosurfactantandBioSolv808BioaugmentationMicrobes,thepetroleumconsumingmicrobesarefavored
in the biological chain and increase in number proportional to those without that special capability. BioSolv 801
Biosurfactantactsinathreestageprocesstoenhancebioremediation.Inthefirststage,nativesandorsoilbacterial
populationsinapetroleumrichenvironmentarestimulatedwiththeBioSolv801growthenhancementsubstrate.At
the same time, BioSolv 801 biodegradable digestive enzymes (natures catalysts) are provided to break down the
petroleumcomplexesintomoredigestiblefragmentsorwhathascometobeknownasmicroscopicmicellulardroplets,
therebyallowingthenativesoilbacteriatobeginconsumingthepetroleumcompounds.
Inthesecondstage,theenlargednativebacterialpopulationundergoesnaturalbioselectivity.Thosebacteriacapableof
utilizingpetroleumcompoundsastheirnatural"foodsource"arepresentedwitharichsourceofdigestiblepetroleum
compounds to continue their growth. Bioselectivity is the primary reason Synergy Services markets BioSolv 808
Bioaugmentation Microbes in order to insure sufficient levels of the bacteria that utilize petroleum products as their
preferredfoodsource.

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ThethirdstageofbiologicalenhancementisoneinwhichsmallamountsofbothBioSolv801BiosurfactantandBioSolv
808 Bioaugmentation Microbes are applied at regular intervals to support, stimulate and build colonies of selective
petroleumconsumingbacteriaandtoprovidesurplusenzymestobreakdownthepetroleumcompoundsforbacterial
digestion.InadditiontothesmallamountsofBiosurfactantandMicrobes,SynergyServicespromotestheutilizationof
BioSolv 817 Biofertilizer in order to enhance the final critical stage in which the natural Gaussian decline in bacteria
populationsiscurtaileduntiltheavailablepetroleumcomplexhasbeenreduced.Oncethepetroleumcompoundshave
beentotallyscavenged,thepetroleumconsumingbacteriarapidlydeclinetotheirnaturalpopulationinthesoilmatrix.
ThefollowingdescribesinmoredetailhowtheSynergyServicescompoundsinteractinpetroleumaffectedsoiland/or
waterbycatalyzingthenaturalbiochemicalreactions:
MICROBIALGROWTH
Whilemicrobesarecapableofutilizingpetroleumhydrocarbonsasanenergysource,theypreferentiallyutilizesimple
sugars (glucose) for rapid growth and energy. Other growth factors required by some microbes include amino acids,
purinesandpyrimidines.SynergyServicesBioSolv817Biofertilizerprovidesarichsourceofthesesimplesugars,amino
acids, and the other related growth factors. Once the glucose energy source is exhausted, those microbes capable of
utilizing petroleum for such growth are naturally favored and their absolute, as well as relative numbers increase
dramatically.
The microbial growth phase is characterized by an initial lag phase while the DNA and enzyme phase develops. The
extracellularenzymesnecessarytobreakdownorganicmoleculesformicrobialgrowtharereadilyavailableinBioSolv
801 Biosurfactant, shortening the lag phase. As soon as the basic requirements for enzyme and DNA production are
met,thesoilmicrobesenteraphasewheretheexponentialgrowthdevelops.Duringthisrapidgrowthphase,microbes
areparticularlysensitivetoadverseconditions,andthusrespondwellwiththepresenceofBioSolv817Biofertilizer.By
continuallyaddingfreshnutrients,microbialgrowthishealthierandthemonitoringofsuchwithvariousfieldtestsisso
important throughout the process. These include pH, byproduct gas analysis (ASTM lengthofstain vials), oxygen
uptake,watersaturation,andconsistentmicroscopicreviewofmicrobialactivity.
ENZYMEINDUCTION
BioSolv 801 provides a rich mixture of extracellular enzymes capable of initiating and catalyzing the breakdown of a
wide variety of petroleum compounds. Extracellular enzymes, which initiate the metabolic (oxidative) process for
biodegradation of petroleum compounds, are produced by certain microbes. In the first step of such oxidation, these
extracellular enzymes instantaneously break off two carbon units from the saturated hydrocarbon chains, which are
typicalofmostpetroleumcompounds.Thetransformedpetroleummoleculeisthenreleasedfromtheenzyme,whichis
thenfreedtoreactwithotherpetroleummoleculesaretransferredintothemicrobeforitsownmetabolicprocess.The
microbes "recognize" from whence the food source originated and alters its own enzyme production to attack the
identifiedfoodsource(inthiscasethepetroleummolecules)directly.Thisiscalled"enzymeinduction".Suchenzyme
inductionintheenlargedsoilbacterialcommunityresultsinanexponentialincreaseinpetroleumbacterialgrowth.
COMETABOLISMANDSECONDARYSUBSTRATEACTIVITY
BioSolv 801 further enhances the cleanup process through cometabolism and secondary substrate activity. Co
metabolicbreakdownoccursasaresultofanontargetcompoundbeinginthepresenceofthehighenergymetabolic
field created with BioSolv 801. This highly structured metabolic process of petroleum compounds is quite capable of
altering nontarget organic compounds and transforming them into readily oxidizable and nontoxic forms which can
then be metabolized by the soil microbial consortium. As the target contaminant decreases below a critical
concentration, microbial populations decrease and the reaction slows or stops completely. This explains why most
bioremediationsoilventingoperationsceasetobeeffectiveasthepetroleumcompoundconcentrationsdecrease.By
maintainingarichmicrobialpopulationwithBioSolv817Biofertilizerscarbonandfoodsources,evensmallresidualsof

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petroleumcompoundscontinuetobemetabolizedtoverylowconcentrations.Thissecondarysubstrateenhancement
by the use of Synergy Services bioremediation compounds allow for a costeffective treatment of large volumes of
petroleumaffectedsoilwithlowhydrocarbonconcentrations.
MOBILIZATIONOFPETROLEUMCOMPOUNDSFORIMPROVEDMICROBIALACTION
BioSolv801Biosurfactantcontainsorganicbiodegradablebioemulsifiers(surfactants)tobreakdowntheadsorptionof
petroleumontosoilparticles.Thisallowsthehydrocarbonstomovemorefreelythroughthesoilporeswheretheless
mobilemicrobialcommunitylives.Thebioemulsifiersbreakdownmacroscopicclumpsofpetroleumintosmallerunits
withincreasedsurfaceareaformicrobialeffect.Thesurfactantactionalsohelpsthenutrientsinmigratethroughthe
soilmatrixmoreeasily.

OPERATIONALOVERVIEW:
Therearemultiplefactorswhichdetermine thespeedandcompletionof thebreakdownofpetroleumcompoundsin
bothsoilandwater.Theaccelerationofthenaturallyoccurringmicroorganicmetabolicandenzymeproductionprocess
isaccomplishedbywayofdeliveringtothenaturallyoccurringbacteriapresentinthesoilanoptimalenvironmentand
foodsource.Thevariouscomponentsnecessarytothisprocessinclude:
Bacteriaandbacterialmobilitywithinthematrix.
Oxygeninsufficientquantitytosupportmetabolism
Moisturecontentcontrol
Temperaturecontrol
pHcontrol
Soilpropertiesclaycontent,porosity,permeability,presenceofothertoxins
The environmental factors must be continually monitored throughout the treatment process in order to create the
optimalbreakdownprocess.Addressingalloftheabovevariablesprovidesasoundbasisforasuccessfulproject.
ENGINEERINGMANAGEMENTOFBIOREMEDIATION
Under normal enhanced soil remediation operations, an aliquot of BioSolv 801 Biosurfactant and water is moved
throughthesoilinaweeklypulsedfashionunderaerobicconditions.Wateriscriticalformetabolicactivity(growthand
reproduction)ofthemicrobialcommunityaswellashydrolysis.Idealmoisturecontentisusuallyat20%to50%offield
soilholdingcapacity.
Sincethevastmajorityofpetroleumbreakdownisperformedbybacteriathatutilizeoxygenintheirmetabolicprocess,
itisnecessarytoensurethatoxygenisavailableinsufficientquantities.Thisisprovidedintheuppersoillayersbymixing
BioSolv801Biosurfactantwithdomesticwater,oxygenatedaquiferwater,and/orairspargedwater.Inextremecases,
asmalldoseofhydrogenperoxidecanbeaddedtotheinjectionsolution.
If the petroleum compounds have penetrated deep into the soil and the conditions are anaerobic, a combination of
BioSolv801Biosurfactant,oxygenatedwaterandothersourcesofoxygencanbeinjected.MovementofBioSolv801
Biosurfactant,waterandoxygenthroughthedeeplayersofthevadosezoneandbeyonddependonsoundengineering
studiestodevelopaplantoutilizeSynergyServicestechnologytoitsfullestpotential.
BioSolv 801 Biosurfactant is provided in a concentrated form. One gallon of the concentrate is used to assist the
bioremediationofapproximatelyfourteencubicyardsofpetroleumaffectedsandorsoil.A1%solution(onegallon/99
gallons water) of BioSolv 801 Biosurfactant is applied to the affected soil in periodic applications in order that the
moisturecontentofthebioremediationbedneedstobemaintainedat20%to50%fieldcapacitybetweenBioSolv801

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Biosurfactant applicationsforsuccessfultreatment.Inaddition,theuseofBioSolv808BioaugmentationMicrobesand
BioSolv817Biofertilizerwillonlyservetospeedandcompletetheremediationprocess.
EXAMPLE
A property has 1,000 cubic yards of petroleumaffected soil. The soil has been placed in a lined berm, 18 inches high
withasoildepthof14inches.UsingtheratioofonegallonofBioSolv801Biosurfactantper14cubicyards,71gallonsof
BioSolv801Biosurfactantwillbeneeded.Eachweek,20%oftheBioSolv801Biosurfactant(14gallons)isappliedina
singledose,dilutedfirstwith1,386gallonsofwater.
Forthefirstapplication,applying28gallonsofBioSolv801Biosurfactantdilutedinamixingtankwith2,772gallonsof
water(1%solution)andthenappliedtothebioremediationbedslowlyenoughforpercolationthroughthesoil.Ifthe
surfaceisimpermeable,tillwithamechanicalorhandrakeisconductedfirsttoincreasepermeability.Theadditional
application of BioSolv 808 Bioremediation Microbes during the tilling, disking, or soil breakup process will greatly
enhanceandspeedthewholeprocess.SynergyServicesrecommendsaugmentingtheprocesswith1poundofBioSolv
808 Bioremediation Microbes per 14 cubic yards of contaminated soil. The addition of microbes can also be broken
downintofourorfivesmallerapplicationstosupplythebioselectivityprocessthroughouttheremediationproject.For
theremainderofthefirstweek,themoisturecontentismaintainedbyaddingoxygenatedwaterwiththeadditionof
BioSolv 817 Biofertilizer at the rate of 1 gallon per 100 gallons of water (1% solution). No additional BioSolv 801
Biosurfactantisused.Attheendoftheweek,thesoilmaybeturnedortilledtoaugmentoxygenation,ifwarranted.
For each subsequent week of treatment, 14 gallons of BioSolv801 Biosurfactant, diluted with 1,386 gallons of water
(1%solution),isappliedfollowedwithmaintenanceofthemoisturecontent.
Inmanycases,themostrapidandcostefficientmethodoftreatmentistospreadthepetroleumaffectedsoilonthe
surface in single layer 14inch lifts and treated under optimal aerobic conditions. However, one can move a weekly
aliquotofBioSolv801Biosurfactantladenwaterand/orairthroughalargerexsitusoilpile.SlottedPVCpipeinstalled
in manifolded grids throughout larger piles can be used to maintain appropriate application of Synergy Services
Biologicals,waterandoxygen.
A sound engineering plan must be developed for the unique conditions existing at each site, taking into account the
manyvariablesinvolvedinmovingthealiquotofBioSolv801Biosurfactant,waterandoxygenthroughthepetroleum
plume.Insomecases,petroleumaffectedgroundwatercanberecycledthroughthebioremediationbedinconjunction
withtheBioSolv801Biosurfactant,therebyyieldinggroundwatercleaningsimultaneously.BioSolv801Biosurfactant
has been used in conjunction with many other remediation technologies for performance enhancement. These
applicationsshouldbecoordinatedcloselywithSynergyServicestoensuresuccessfulapplication.
TOXICOLOGY
TheingredientsofSynergyServicesbiologicalshavebeensubjectedtotoxicologicaltestinginmammalsandmarinelife
toassessitssafetyinintendedfielduses.Undernormaloperatingparameters,BioSolv801Biosurfactantisappliedata
dosageof0.100mg/l.Ofallthemultifacetedorganicingredients,whenindividuallytested,theharshestcomponentin
marinelifestudiesdemonstratedthatthe48hourmediallethalconcentration(LC50)determinedonbrineshrimpwas
greaterthan100mg/landthe96hourLC50determinedonfishwasfoundtobe5.3mg/l.Ofthatcomponentatthe4
percent by weight concentration contained in the proprietary formulation these calculations can be exponentially
increasedaccordingly.
PROJECTRESULTS
Resultsofselectedbenchtests,pilotscaletestsandactualfieldprojectimplementationareappended.

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TESTPROTOCOL:
I.TestProtocolMethodology
Thetestsoilcontaminatedwithpetroleumproductshouldfirstbecharacterizedandthendividedintothreeparts,one
for control (blank), one for testing BioSolv 801 Biosurfactant alone, and one for testing BioSolv 801 Biosurfactant,
BioSolv808BioaugmentationMicrobes,andBioSolv817Biofertilizer.
Wheninthefullscalemode,itisstilladvisedtoreserveasmallcontrolareasorelativedegradationobservationcanbe
correlated.Inthelaboratory,however,eachsoiltestvolumeshouldbeplacedinopentoppedtestpanstodepthofsix
inches.ThecalculatedvolumeofpredilutedBioSolv801Biosurfactantsolutionshouldbeaddedtothetwotreatedtest
bedsandanequivalentamountofwatershouldbeaddedtothecontrolbed.Thenallthreetestbedsshouldbebrought
toapproximately30percentmoisturebyaddingwater,withtheexceptionofthethirdtestsamplethatwillrequirea
controlled portion of BioSolv 808 Bioaugmentation Microbes (powdered microbes) and the moisture content can be
increased with a 1% solution of BioSolv 817 Biofertilizer in order to determine the combined technologys efficacy in
comparison to water alone, and water mixed with BioSolv 801 Biosurfactant. This addition of water and Synergy
Servicescompoundsshouldbecontinuedinfiveequalweeklydosesoverafiveweekperiodinordertoreplicatefield
conditions.Onadailybasis,inallthreematrices,moisturelevelsshouldbemaintainedat20to25percentatalltimes,
oranamounttomaintainmoistconditionswithoutproducingleachateorstandingwater.Inactualfieldconditions,the
sameprotocolshouldbeobserved,buttheturningofthesoilmaybeconductedonalessfrequentbasis,dependingon
thesoilcharacteristicsandrainconditions.
Abacterialcountisrecommendedpriortocommencementoftestingtocheckforsoilsterility.Additionally,thebacteria
inthesoilshouldhavebeenexposedtopetroleumhydrocarbons,allowingsomeformstoalreadybeacclimated.This
shortensthelagtimeforexponentialbacterialgrowthoncetheBioSolv801Biosurfactanthasbeenadded.Ifthesoilis
devoid of bacteria, Synergy Services recommends the application of 2 pounds of BioSolv 808 Bioaugmentation
Microbesper14cubicfeetofcontaminatedsoil.
II.TestProtocolProductUtilization
One gallon of BioSolv 801 Biosurfactant concentrate will augment treatment of fourteen cubic yards of
petroleumaffectedsoil.
OnepartofBioSolv801Biosurfactantconcentrate(1%)shouldbepremixedwith99partswater(99%).
For each fourteen cubic yards of soil to be treated, the one gallon of BioSolv 801 Biosurfactant concentrate
(mixedwithoxygenatedwater)shouldbeappliedonefifthatatimeforfiveweeklydoses.
SynergyServicesrecommendsapplyingBioSolv817Biofertilizerwheneverwaterisaddedtothebioremediation
sitetospeedbacterialgrowthandthebiodegradationprocess.Ifspeedisntapriorityinyourprocess,then
youmayeliminatethisaugmentationproduct.
Inordertofullybiodegradetheremediationsite,SynergyServicesrecommendstheadditionoftheBioSolv808
Bioaugmentation Microbes at the rate of one pound per fourteen cubic yards of petroleumaffected soil,
regardless of the current microbial count. If the site is found to contain little or no petroleumacclimated
microbes, then two pounds of BioSolv 808 Bioaugmentation Microbes should be added per fourteen cubic
yardsofsoil.
In order to promote bioselectivity and stay on the upside of the Gaussian decline in microbial activity and
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promotetreatmentclosure,onecubicyard(0.756m )ofsoil,useatotalofoneounce(32.4grams)ofBioSolv
808 Bioaugmentation Microbes. You may find the best results by dividing these microbe aliquots over five
weeklydosesorinthiscase6.48gramspercubicyd.ofsoilperweek(5X).
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ManysiteswillfindusingBioSolv801Biosurfactantwithoutanyothertreatmentcompoundandhavedelivered
excellentresultsduetothepropernutritionandmicellularbreakdownofthelargeoilparticles.
It is imperative that the remediation site receive proper soil handling for particle surface area treatment and
oxygenation.Waterisalsorequiredforphasetransmittalofbionutritionandaugmentationofmuchrequired
oxygeninordertomaintainahealthyBOD/COD(measuredbyoxygenuptakeinthefield).

MAJORGENERAOFOILDEGRADINGBACTERIAANDFUNGI

BACTERIA FUNGI
Achromobacter Coryneforms Allescheria Penicillium
Acinetobacher Erwinia Asperglilus Rhodosporidium
Actinomyces Flavobacterium Aurebasidium Rhodotorula
Aeromonas Klebsiella Botrytis Sccharomyces
Alcaligenes Lactobacillus Candida Saccharomycopisis
Arthrobacter Leucothrix Cephalosporium Scopulariopsis
Bacillus Moraxella Cladosporium Sporobolomyces
Beneckea Nocardia Cumminghamella Torulopsis
Achromobacter Peptococcus Debaromyces Trichoderma
Acinetobacher Pseudomonas Fusarium Trichosporon
Actinomyces Sarcina Gonytrichum
Aeromonas Spherotilus Hansenula
Alcaligenes Spirillum Helminthosporium
Arthrobacter Streptomyces Mucor
Bacillus Vibrio Oldiodendrum
Beneckea Xanthomyces Paecylomyces

Brevebacterium Phialophora

GENERALCOMMENTS:
Historically, bioremediation of petroleum based compound incidents has been limited by the uncertainty of time and
completenessofthereactions.Bacterialbasedenhancementproductshavehelpedinlimitedapplications,buthavenot
beencompletelysuccessfulinallcases.Recently,nonpathogenenzymeagentshavecomeontothecommercialmarket
which,combinedwithairinjectiontechnology,enablecompaniestolookatbioremediation(bothinsituandexsitu)asa
fasttrackalternative.
Soilandgroundwaterremediationmaybeacceleratedusingablendofbioremediationenhancingagentstostimulate
nativemicrobestoreducebiologicalreactionkineticstolessthaneightweeks,andsometimeslessthanfourweeks.
Mostsoil,groundwater,andfreeproductcontaminatedsiteshavenativemicroorganismspresentthatareacclimatedto
destroy hydrocarbons. To accelerate the breakdown process, addition of a blend of enzymes, amino acids, glucose,
oxygen,andmoisturecontrolisallthatisrequiredtocompressbioremediationtimesfromyearstoweeks.
CopyrightedInformation
AllinformationcontainedhereinisthepropertyofSynergyServicesandwaswrittenfornoncommercialuseonlyasatrainingtool.UnlesspermissionisgrantedbySynergyServices,Inc.,
thisTechnologyPrimershallnotbeduplicatedorusedforcommercialpurposes.

DisclaimerofLiability:SynergyServices,Inc.(hereinknownasSynergy)warrantstopurchasers,butnothirdpartiesorothers,thespecificationsfortheproductshallfallwithagenerally
recognized rangefor typical properties established bySynergy when the product departs from Synergys pointof origin and that any services shall only be performed in WARRANTY OF
MERCHANTABLITILYORFITNESSFORAPARTICULARPURPOSE,REGARDINGANYSERVICESPERFORMEDORPRODUCTSUPPLIED.SynergywillgivepurchaserthebenefitofSynergysbest
judgmentinmakinginterpretationsofdata,butdoesnotguaranteetheaccuracyorcorrectnessofsuchinterpretations.Synergysrecommendationscontainedhereinareadvisoryonlyand
withoutrepresentationastotheresults.Synergyshallnotbeliableforanyindirect,special,punitive,exemplaryorconsequentialdamagesorlossesfromanycausewhatsoeverincluding
butnotlimitedtoitsnegligence.