С - Danish Technological Institute

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С - Danish Technological Institute

The resident communities include a variety of anaerobicmicroorganisms:Fermentative, sulfate-reducers, manganese and iron-reducers, acetogensand methanogens.Aerobic bacteria observed in oil fields are believed to beexogenous contaminants.Scheme of oil field exploited with water-flooding1 12 21 – near-bottom zone of injection well 2 – production well

PetroleumAerobic +O 2Surfactants, CO 2 , oxidizingpolysaccharidesHd Hydrocarbon-Microbialtransformation ofpetroleumaccompaniedwith productionof oil-releasingagents in a waterfloodedoil fieldBiomass, Fatty acids, alcohols,Exometabolites aromatic compounds_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _AnaerobicSolvents, gases,lower fatty acidsFermentativebacteriaPropionate, butyrate,pyruvate, ethanol, propanolSynthrophicbacteriaFe3+-H 2 , CO 2 , formate, reducingacetate, methanolSulfate-Methanogens reducingFe 3+ 2-+SO 4CH 4 , CO 2 Fe 2+ , CO 2 H 2 S, CO 2The field tests ofthe biotechnologygave positiveresults; over750000 tons of oilwas additionallyrecovered.

Methods of investigation1. Analytical (chemistry of water, oil, gases)2. Microbiological methodsUse of selective media for:• Aerobic organotrophs• Hydrocarbon-oxidizing bacteria• Fermentative bacteria• Sulfate-reducing bacteria• Methanogens3. Methods of Polyphase TaxonomyMorphology, physiology and phylogenetic position ofmicroorganisms by 16S rRNA gene sequencing4. Radioisotope methods• Sulfate reduction – Na352SO 4• Lithotrophic methanogenesis – NaH 14 CO 3• Aceticlastic methanogenesis - 14 CH 3-COONaSampling5. Molecular biology techniques• DNA and RNA extraction• Amplification and cloning of 16S rRNA genes• Sequencing and phylogenetic identification

CHARACTERISTICS OF THE KONGDIAN AREA OF THEDAGANG OIL FIELDDepthTemperature1206 – 1434 m59 o CpH 7.1-7.6Salinity it 06% 0.6 Water cut 95%22productionwells11injectionwellsLayout of injection and production wellsat the experimental site of theKongdian oil field

• Near-bottom zone*:Chemical and bacteriological parameters of theKongdian bed before the trialCharacteristic• Aerobic bacteria

Aerobic bacteria isolated from the Dagang oil field•34 aerobic strains Geobacillusjurassicus DS1 Т•45-65 °С:Geobacillus(G. subterraneus, G. pallidus,G. stearothermophilus,G. thermoglucosidasius)ThermoactinomycesAbundance,%•20-45 °С:24680Bacillus,Oceanobacillus,8Micrococcus,64Cellulomonas,2Pseudomonas0AcinetobacterAbundance, %101214С12GLC tracing of crude oil satrated hydrocarbons afterexposure to degradation by G. jurassicus DS2С 10С 12С 14С 16С 18С 20С 22С 24С 26С 28С 30С 32С 34С 36Numbers of carbonsweathered oil, control incubated at 60 0 CС14С16С18С20Branched alkanesС22С24Number of carbonsС26С28С30С32Unbranched alkanes

Diversity of alkB homologs in bacteria of thegenus GeobacillusalkB Nearest alkane 1-monooxygenase according to Nucleotidehomolog BLAST analysis sequencesimilarity, %alkB-geo1 alkB4 from R. erythropolis NRRL B-16531 99.2alkB-geo2 alkB4 from R. erythropolis NRRL B-16531 90.0alkB-geo3 alkB3 from Nocardia sp. H17-1 87.7alkB-geo4 alkB3 from R. erythropolis NRRL B-16531 96.7alkB-geo5 alkB2 from R. erythropolis 50-V 95.4alkB-geo6 alkB2 from R. erythropolis NRRL B- 99.0alkB-geo7 alkB, environmental clone alkG4-35k 69.8alkB-geo8 alkB1 from Rhodococcus sp. Q15 70.00

• Rheological og characteristics acte cs of culture media of aerobicthermophilic bacteria from the Kongdian bedStrain,substrateMicrobialcommunity1098-25 m 3Emulsifyingactivity, %Surface tension,mN/mInterfacialtension*, mN/mViscosity, mPa xs(at 60 °C)Sucrose 50 53.1 19.3 0.68Acetate 30 51.3 19.0 0.66C10-C20paraffin40 51.5 25.1 0.62Crude oil 30 41.9 11.8 0.63Geobacillus jurassicus DS1 ТSucrose 30 45.3 15.5 0.64Acetate t 30 47.3 15.3 061 0.61C10-C20paraffin5 51.3 22.7 0.64Crude oil 30 41.9 12.5 0.60

--Determination of the filtration properties of the collector ofthe Kongdian bed1017-7 1092109464,2 Бк/л1,76 Бк/л1094-1418,3 Бк/л10151017-51,89 Бк/л1013-1115,6 Бк/л13,78 Бк/л1,44 Бк/л1,96 Бк/л1015-110171008-1100801 1050-31017Appearance of tracersin formation watersfrom production wellslocated around theinjection wells1050-31002-160,2Бк/л58,9 Бк/л10501032-189,0 Бк/л36,0 Бк/л1050-111050-2

•Development of a technological scheme of the activation offormation microflora;•Development of a field engineering process for injectingwater-air mixture and mineral salts into the stratum;•Injection of the water-air mixture and nutrients into thestratum.

Monitoring the microbiological,biogeochemical and productioncharacteristics of the oil field in thecourse of the pilot trialApproximately 40 parameters of the fluids from 22production wells, of backflushed water from the injection welland of injection water were monitored for 5 years.

The numbers ofmicroorganisms in formationwater from wells 1015-1 inthe course of the pilot trialThe rate of sulfate reduction information water of the Kongdianbed in the course of the pilot trial1015-1 300lglls number,Ce876543210June e,2000.,2000Dec.,2001July,Jan.,2 2002June,2 002Jan.,20 003FermentMet-H2HOBSulf fate reduction rate, u g S2-/l/day250200150100500Dec2000July2001Jan2002June2002Jan2003Dec2004May2005Dec2005May2006Dec20061017-46310921094-11050-1HOB SRB Met-H2 Met-Acet Ferment1050-1 1094-1 1092 63 1017-4

Peak values of methanogenesis rate in formationwater in the course of the trial(μg of CH 4 L -1 day -1 )45 14004012003530100025800206001540010520001631002-110081008-110121012-11015-110171017-21017-31017-41017-51017-710321032-11050-11050-21050-31092109408 m3 25 m3 1094-1From NaH14CO3 From 14CH3-COONa From NaH14CO3 From 14CH3-COONa

Physicochemical parameters of formation water and gas of the Kongdianbed during the biotechnological trial180140016012001401201008060A c etate, m g / l1000800600400НСО3, mg/l200400Sep., 2003200631002-110081008-110121012-11015-110171017-21017-31017-41017-51 017-710320 32-10 50-150-25 0-394-11Jan., 2002June, 2000Sept. 2003June, 2002July 2001May 200511 01 0101051 0 921010941631002-110081008-110121012-11015-110171017-21017-31017-41017-51017-710321032-11050-11050-21050-3109210941094-10-200 200-400 400-600 600-800 800-1000 1000-1200 1200-1400July 2001 Jan. 2002 June, 2002 Jan. 2003 Sept. 2003 Nov 2004 May 2005Content of HCO 3-and acetate in formation waters of the Kongdianbed in the course of the pilot trial (mg/l)

The rheological characteristicsof formation water from well• Emulsifying activity of formation1008-1 in the course of the pilottrialfluids of the Kongdian bed50454035/mmN30252010 10 102 08- 0-1 1 081015 1-1 0121017-463151050Dec 2000July 2001Jan 2002June, 2002Jan 2003Sept 2003Surface tensionInterfacial tensionInterfacial tension Surface tension • Viscosity of formation waterincreased from 0.70 to 0.86mPa x s.

Gas composition and Biogeochemical Characteristics of theKongdian BedIn the course of the trial:CO 2 increased on 2-4.1 1%;CH 4 increased on 2-5.7 % in thegas of several production wells.• δ 13 C/CH 4 , o / oo (in the gas)-151631 002-110081 008-110121 012-11 015-110171 017-21 017-31 017-41 017-51 017-710321 032-11 050-11 050-21 050-3109210941 094-111111111111111•δ 13 C∑CO 2 + HCO 3- +CO2- -253(in the formation water)Before the trial:+6.4 o / oo ÷-1.5 o / oo-35-45In the course of the trial:+6.4 o / oo ÷ -12.9 o / oo (14 wells)-55+6.4 o / oo ÷ +10.6 o / oo (3 wells)-65Dec 2000 July 2001 Jan 2002 June 2002 Jan 2003 Sept 2003

Additional oil production on the North block of the Kongdianbed as a result of application of MEOR technology in V.2001-V.2007WellNo.Decline formulationAdditionaloil, t1002-1 Q 0 =16.75(1+0.5*0.039t) -2 94951008-1 Q 0 =9.8(1+0.5*0.02t) -2 38921015-1 Q 0 =11(1+0.5*0.021t) -2 52781017-2 Q 5*0 028t) 0 =7.15(1+0.5*0.028t) -2 30521017-3 Q 0 =8.31(1+0.5*0.072t) -2 4461017-7 7 Q =24.11(1+0.5*0.027t) -2 0 0.027t) 19321032-1 Q 0 =31.71(1+0.5*0.02t) -2 58681050-3 Q 0 =5.48(1+0.5*0.027t) -2 50361094-1 Q 0 =5.99(1+0.5*0.031t) -2 1003Total 36002Decline curve of oil productionon the North block in 1997-2007

CONCLUSIONS• It was experimentally proved that the high-temperature oil field is anecosystem in which a biotic community interacts with an abioticmedium so that energy fluxes create a certain trophic structure. Theseenergy fluxes are based on the biotransformation of oil in particulartrophic chains and can be regulated in a goal-directed manner.• The possibility of the application of the MEOR technology based onthe activation of metabolism of the formation microorganisms wasdemonstrated in high-temperature oil field.• As the result of the injection of water-air mixture and mineral nutrientsmicrobial transformation of residual oil was accompanied byaccumulation of bicarbonate b ions, volatile fatty acids, biosurfactants,tand microbial biomass in the stratal water, and of CH 4 and CO 2 in thegas phase.• Microbial metabolites promoted additional oil replacement from thelayer. About 36 thousand tons of oil was recovered additionally on theDagang oil field as a result of MEOR application.

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