LT4522B - Petroleum oil and its products decomposing microorganism arthrobacter globiformis md18 - Google Patents
Petroleum oil and its products decomposing microorganism arthrobacter globiformis md18 Download PDFInfo
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- 244000005700 microbiome Species 0.000 title claims abstract description 29
- 241000186074 Arthrobacter globiformis Species 0.000 title claims abstract description 26
- 239000003208 petroleum Substances 0.000 title claims description 6
- 239000002689 soil Substances 0.000 claims abstract description 45
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 150000001875 compounds Chemical class 0.000 claims abstract 2
- 239000003921 oil Substances 0.000 claims description 20
- 239000000295 fuel oil Substances 0.000 claims description 13
- 239000002609 medium Substances 0.000 claims description 11
- 238000004140 cleaning Methods 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- 239000012531 culture fluid Substances 0.000 claims description 8
- 239000003209 petroleum derivative Substances 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- 235000016709 nutrition Nutrition 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 5
- 239000002002 slurry Substances 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 239000003337 fertilizer Substances 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
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- 240000006909 Tilia x europaea Species 0.000 claims description 2
- 235000011941 Tilia x europaea Nutrition 0.000 claims description 2
- 230000001580 bacterial effect Effects 0.000 claims description 2
- 229910052799 carbon Inorganic materials 0.000 claims description 2
- 230000000694 effects Effects 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 claims description 2
- 239000004571 lime Substances 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 239000010902 straw Substances 0.000 claims description 2
- 239000000758 substrate Substances 0.000 claims description 2
- 238000009423 ventilation Methods 0.000 claims description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims 2
- 239000007921 spray Substances 0.000 claims 2
- 239000007900 aqueous suspension Substances 0.000 claims 1
- 238000007865 diluting Methods 0.000 claims 1
- 235000015097 nutrients Nutrition 0.000 claims 1
- 235000010344 sodium nitrate Nutrition 0.000 claims 1
- 239000000243 solution Substances 0.000 claims 1
- 238000005507 spraying Methods 0.000 claims 1
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 208000034809 Product contamination Diseases 0.000 abstract 1
- 230000015556 catabolic process Effects 0.000 description 11
- 238000006731 degradation reaction Methods 0.000 description 10
- MWPLVEDNUUSJAV-UHFFFAOYSA-N anthracene Chemical compound C1=CC=CC2=CC3=CC=CC=C3C=C21 MWPLVEDNUUSJAV-UHFFFAOYSA-N 0.000 description 8
- NNBZCPXTIHJBJL-UHFFFAOYSA-N decalin Chemical compound C1CCCC2CCCCC21 NNBZCPXTIHJBJL-UHFFFAOYSA-N 0.000 description 8
- 241000589149 Azotobacter vinelandii Species 0.000 description 7
- 229930195733 hydrocarbon Natural products 0.000 description 7
- 150000002430 hydrocarbons Chemical class 0.000 description 7
- 238000005067 remediation Methods 0.000 description 7
- 239000004215 Carbon black (E152) Substances 0.000 description 6
- RZJRJXONCZWCBN-UHFFFAOYSA-N octadecane Chemical compound CCCCCCCCCCCCCCCCCC RZJRJXONCZWCBN-UHFFFAOYSA-N 0.000 description 6
- 125000003118 aryl group Chemical group 0.000 description 4
- 239000003344 environmental pollutant Substances 0.000 description 4
- 230000035764 nutrition Effects 0.000 description 4
- 231100000719 pollutant Toxicity 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- PXXNTAGJWPJAGM-UHFFFAOYSA-N vertaline Natural products C1C2C=3C=C(OC)C(OC)=CC=3OC(C=C3)=CC=C3CCC(=O)OC1CC1N2CCCC1 PXXNTAGJWPJAGM-UHFFFAOYSA-N 0.000 description 4
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 239000000356 contaminant Substances 0.000 description 3
- 238000000354 decomposition reaction Methods 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 229940038384 octadecane Drugs 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- UFWIBTONFRDIAS-UHFFFAOYSA-N Naphthalene Chemical compound C1=CC=CC2=CC=CC=C21 UFWIBTONFRDIAS-UHFFFAOYSA-N 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 239000010692 aromatic oil Substances 0.000 description 2
- DIOQZVSQGTUSAI-UHFFFAOYSA-N decane Chemical compound CCCCCCCCCC DIOQZVSQGTUSAI-UHFFFAOYSA-N 0.000 description 2
- DCAYPVUWAIABOU-UHFFFAOYSA-N hexadecane Chemical compound CCCCCCCCCCCCCCCC DCAYPVUWAIABOU-UHFFFAOYSA-N 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 239000012266 salt solution Substances 0.000 description 2
- 241000186063 Arthrobacter Species 0.000 description 1
- 241000589151 Azotobacter Species 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241000549556 Nanos Species 0.000 description 1
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 241000589776 Pseudomonas putida Species 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
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- 150000001491 aromatic compounds Chemical class 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
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- 239000000571 coke Substances 0.000 description 1
- 238000009264 composting Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000003305 oil spill Substances 0.000 description 1
- 238000006395 oxidase reaction Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000010690 paraffinic oil Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
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- 208000024891 symptom Diseases 0.000 description 1
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- 239000008096 xylene Substances 0.000 description 1
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- Processing Of Solid Wastes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
Description
Išradimas priklauso mikrobiologijos bei aplinkos apsaugos sritims, būtent, naftą ir naftos produktus skaidantiems mikroorganizmams. Išradimas gali būti panaudotas vandens bei įvairių dirvožemių, užterštų naftos produktais, ypač sunkiosiomis jo frakcijomis, biologiniam valymui.The invention relates to the fields of microbiology and environmental protection, namely, microorganisms which break down oil and petroleum products. The invention can be used for biological treatment of water and various soils contaminated with petroleum products, in particular its heavy fractions.
Žinomas JAV patente Nr. 3813316 aprašytas Pseudomonas aeroginosa PAO kamienas, kuris turi plazmidės CAM, OCT, NAH ir SAL su angliavandenilių utilizacijos genais ir naudojamas naftos produktų skaidymui.Known in U.S. Pat. No. 3813316 describes a strain of Pseudomonas aeroginosa PAO which contains the plasmids CAM, OCT, NAH and SAL with hydrocarbon utilization genes and is used for the degradation of petroleum products.
Šio kamieno trūkumas - labai lėtas naftos degradacijos procesas.The disadvantage of this strain is the very slow process of oil degradation.
Žinomas išradimo aprašyme (pagal buv. TSRS a.l. Nr.612958) aprašytas Pseudomonas aeruginosa kamienas, turintis plazmidės OCT+, CAM+ ir NAH+, naudojamas biologiniam vandens, užteršto naftos produktais, valymui.A strain of Pseudomonas aeruginosa containing the plasmids OCT + , CAM + and NAH + as used in biological purification of water contaminated with petroleum products is known in the specification (ex USSR 612958).
Šio kamieno trūkumas yra siauras jo veikimo spektras.The disadvantage of this strain is its narrow range of action.
Žinomas išradimo aprašyme (pagal buv. TSRS a.l. Nr.1076446) aprašytas Pseudomonas putida 36 kamienas, skaidantis naftą ir jos produktus vandenyje ir dirvožemyje. Šis kamienas išskirtas Tiumenės srityje iš Belozersko naftos radimvietės 13100 gręžinio.A strain of Pseudomonas putida 36, which decomposes petroleum and its products in water and soil, is known from the description of the invention (ex USSR al.l. No. 1076446). This strain was extracted from the 13100 borehole at the Belozersk oilfield in the Tyumen region.
Šio kamieno trukumas - lėtas aromatinių junginių skaidymas. Per 30 parų suskaido 74,538 parafininės naftos, o aromatinės - 68 % nuo pradinio kiekio.The disadvantage of this strain is the slow degradation of aromatic compounds. 74,538 paraffinic oils decompose in 30 days and aromatic 68% of initial volume.
Artimiausias savo veikimu siūlomam kamienui yra kamienas, aprašytas LR patenteThe closest to the proposed strain is the strain described in the LR patent
Nr.3111 B, Azotobacter vinelandii 21, išskirtas iš užteršto dirvožemio sunkiosiomis naftos frakcijomis po tanklaivio avarijos ir siūlomas naudoti gėlo ir sūraus vandens bei dirvos valymui nuo naftos ir jos produktų teršalų. Kamienas deponuotas UAB “Biocentras” mikroorganizmų muziejaus kolekcijoje, kolekcinis numeris BC-21.No. 3111 B, Azotobacter vinelandii 21, isolated from contaminated soil by heavy oil fractions after a tanker accident and proposed for use in the cleaning of fresh and salt water and soil from contaminants of petroleum and oil products. The strain is deposited in the collection of the Microorganism Museum of Biocentras UAB, collection number BC-21.
Šio kamieno trūkumas - gana lėtas naftos ir sunkiųjų jos frakcijų skaidymas . Per 21 parą šis kamienas suskaidė iki 75,6% parafininės naftos ir iki 80,2% aromatinės naftos nuo pradinio kiekio.The disadvantage of this strain is the relatively slow decomposition of oil and its heavy fractions. Within 21 days, this strain decomposed to 75.6% of paraffin oil and up to 80.2% of aromatic oil from its original content.
Išradimo tikslas - pagreitinti sunkiųjų naftos frakcijų skaidymą.The object of the invention is to accelerate the decomposition of heavy oil fractions.
Siūlomas Arthrobacter globiformis MD18 kamienas, išskirtas iš mazutu užteršto Vilniaus naftos bazės dirvožemio. Kamienas deponuotas Botanikos instituto Biodestruktorių tyrimo laboratorijoje, kolekcinis numeris B-84.Proposed strain of Arthrobacter globiformis MD18 isolated from oil-polluted soil of Vilnius oil base. The strain is deposited in the Biodestructors Research Laboratory of the Botanical Institute, Collecting Number B-84.
Arthrobacter globiformis MD18 kamienas gautas panaudojant mikrobiologinę kultūrų skyrimo metodiką, būtent užteržtas mazutu dirvožemio mėginys buvo įbertas į Capeko terpę su skaidomu substratu (mazutu) ir aeruojant kultivuojamas 4-6 paras apie 30°C temperatūroje, gauta kultūra praskiedimo metodu buvo išsėta ant standžios Capeko terpės su mazutu, kaip vienintelės anglies ir energijos šaltinio, po to vėl kultivuojama 5 - 6 paras apie 30°C temperatūroje. Užaugusios izoliuotos bakterijų kolonijos išsėjamos i mėgintuvėlius ant įžambaus Čapeko terpės agaro su mazutu. Po to buvo lyginamas išskirtų kultūrų aktyvumas ir išrenkamas aktyviausiai skaidantis naftos produktus kamienas. Atrinktasis aktyviausias mikroorganizmo kamienas vadovaujantis Bergey’o apibūdintoju (Murray R.G.F. et ai (eds. board), 1984-1989: Bergeys Manual of Systematic Bacteriology 9th ed, The Williams and Wilkins Co., Baltimore) apibūdintas kaip Artfirobacterglobiformis MD18.The strain of Arthrobacter globiformis MD18 was obtained by microbiological culture isolation, namely, the soil sample contaminated with fuel oil was poured into Capek medium with degradable substrate (fuel oil) and cultured for 4-6 days at about 30 ° C, and the resulting culture was diluted onto rigid Capek medium. with fuel oil as the sole source of carbon and energy, then cultivated again for 5 to 6 days at about 30 ° C. Overgrown isolated bacterial colonies are inoculated into tubes on slanting Chapek medium agar with fuel oil. This was followed by comparing the activity of isolated cultures and selecting the strain most active in the decomposition of petroleum products. The selected most active strain of the microorganism is described as Artfirobacterglobiformis MD18 according to Bergey's characterist (Murray R.G.F. et al. (Eds. Board), 1984-1989: Bergeys Manual of Systematic Bacteriology 9th ed., The Williams and Wilkins Co., Baltimore).
Siūlomas mikroorganizmo kamienas pasižymi šiomis savybėmis:The proposed micro-organism strain has the following properties:
Morfologinės savybės. Ląstelės jaunoje kultūroje yra 1,0 - 1,2 x 2,3 - 2,9 ųm dydžio, o senose kultūrose jos virsta kokais, kurie sudaro grandinėles. Morfologiniai formos kitimai priklauso nuo amžiaus ir kultivavimo sąlygų. Nesudaro endosporų, neturi kapsulių bei neturi žiuželių. Gramteigiamos.Morphological properties. Cells in young culture are 1.0 - 1.2 x 2.3 - 2.9 µm in size and in old cultures they turn into coke forming chains. Morphological variations in shape depend on age and culture conditions. It does not form endospores, does not have capsules and does not have blisters. Gram positive.
Kultūriniai požymiai. Ant mėsos peptono agaro (MPA) užauga per 48 vai. ir yra balsvos, netaisyklingos, nelygaus krašto, iškilo profilio, blizgančios kolonijos 2-3 mm skersmens. Ant sintetinės terpės su nafta užauga per 24 vai. sudarydamos smulkias baltas kolonijas. Mėsos peptono buljone (MPB) plėvelės nesudaro, duoda drumstumą ir nuosėdas.Cultural attributes. It takes 48 hours to grow on meat peptone agar (MPA). and are whitish, irregular, uneven edge, raised profile, glossy colony 2-3 mm in diameter. It grows on synthetic medium with oil in 24 hours. forming fine white colonies. Meat peptone broth (MPB) does not film, gives turbidity and sediment.
Fiziologinės savybės. Aerobinės. Katalazės reakcija yra teigiama, o oksidazės neigiama. Tipiški mezofilai, auga nuo 4° iki 40°C, optimali temperatūra 28°-30° C. Krakmolo nehidrolizuoja, vykdo denitrifika'ciją, želatinos neskystina, naudoja gliukozę, fruktozę, sacharozę bei manitą. Auga ant Ešbio terpės.Physiological properties. Aerobic. The catalase reaction is positive and the oxidase reaction is negative. Typical mesophiles grow from 4 ° to 40 ° C, optimum temperature from 28 ° to 30 ° C. Starch does not hydrolyze, undergo denitrification, does not liquefy gelatin, uses glucose, fructose, sucrose and mannitol. Grows on Ashbee medium.
Auga ant sintetinės terpės su angliavandeniliais: heksanu, dekanu, heksadekanu, oktadekanu, antracenu, toluolenu, ksilolenu, naftalenu, dekalinu, parafinu bei asfaltenais.It grows on synthetic media containing hydrocarbons: hexane, decane, hexadecane, octadecane, anthracene, toluene, xylene, naphthalene, decalin, paraffin and asphaltenes.
Siūlomo Arthrobacter globiformis MD18 ir žinomo Azotobacter vinelandii 21 kamienų naftą degraduojančių savybių palyginimas.Comparison of the oil-degrading properties of the proposed Arthrobacter globiformis MD18 and known Azotobacter vinelandii 21 strains.
Požymiai Arthrobacter Azotobacter globiformis MD 18 vinelandii 21Symptoms of Arthrobacter Azotobacter globiformis MD 18 vinelandii 21
Naftos degradacija (%)Oil degradation (%)
Aukščiau pateiktų duomenų apibendrinimas rodo, kad siūlomas kamienas per 18 parų parafininę naftą suskaidė iki 77,2%, aromatinę iki 82,4%, o Azotobacter vinelandii 21 per 21 parą suskaidė atitinkamai iki 75,7% ir 80,2% nuo pradinio kiekio. Per 20 parų Arthrobacter globiformis MD18 suskaidė naftos sunkiųjų frakcijų iki 75,6%, o Azotobacter vinelandii 21- 72,5 % nuo pradinio kiekio. Vandenyje Arthobacter globiformis MD18 suskaidė 78,4% naftos, o Azotobacter vinelandii 21 - 77,4% nuo pradinio kiekio. Arthrobacter globiformis MD18 suskaidė 74,9% antraceno, 44,5% dekalino, 42,7% oktadekano ir 29,7% asfaltenų, tuo tarpu Azotobacter vinelandii 21 72,6%, 44,4%, 36,7%, 27,5% atitinkamai nuo pradinio kiekio.The summary of the above data shows that the proposed strain decomposed to 77.2%, aromatic to 82.4% within 18 days, and Azotobacter vinelandii 21 to 75.7% and 80.2% from the initial amount in 21 days, respectively. . Within 20 days, Arthrobacter globiformis MD18 degraded heavy oil fractions to 75.6% and Azotobacter vinelandii 21-72.5% from baseline. In water, Arthobacter globiformis MD18 degraded 78.4% of the oil and Azotobacter vinelandii 21 77.4% of the original amount. Arthrobacter globiformis MD18 cleaved 74.9% anthracene, 44.5% decalin, 42.7% octadecane and 29.7% asphaltenes, whereas Azotobacter vinelandii 21 72.6%, 44.4%, 36.7%, 27, 5% from the initial quantity respectively.
Arthrobacter globiformis MD18 kamieno panaudojimo pavyzdžiai.Examples of use of Arthrobacter globiformis MD18 strain.
pavyzdys.example.
Dirvos, ant kurios išsipylė nafta, vietinis valymas. Nafta užteršto dirvožemio plotas 200 m2, teršalų kiekis 2 kg/1 m2 . Bendras teršalų kiekis 400 kg. Dirvos valymui paruošia Arthrobacter globiformis MD18 suspensiją: fermentatoriuje Čapeko terpėje išaugina 100 1 kultūrinio skysčio (8-10 g/l sausos masės). Išaugintą kultūrinį skystį praskiedžia ΙχΙΟ31 mitybai reikalingų druskų vandeniniu tirpalu, pavyzdžiui, NaNOs 4g/l, K2HPO4 - 0,6 g/l, Na2HPO4 - 0,6 g/l. Gautą suspensiją išpurškia ant užteršto dirvožemio paviršiaus, po to dirvožemį perkasa ir išpurena. Valymo proceso eigoje dirvos sluoksnį purena periodiškai 1 kartą per savaitę. Natūraliomis sąlygomis, esant oro temperatūrai 15 - 22° C, dirvos išsivalymo procesas trunka 2 - 3 mėnesius. Dirvožemio -išsivalymo kokybę nustato' specializuotoje ekologinės tarnybos laboratorijoje atliekant laboratorinę dirvos mėginio analizę pagal likutinę angliavandenilių koncentracjją. · pavyzdys.Local cleaning of oil spill. Oil contaminated soil area 200 m 2 , pollutant content 2 kg / 1 m 2 . Total emissions 400 kg. Prepares Arthrobacter globiformis MD18 suspension for soil cleaning by cultivating 100 l of culture fluid (8-10 g / l dry weight) in a Chapek fermenter. Harvested culture fluid is diluted with 3 1 ΙχΙΟ nutrition salts with an aqueous solution such as nano 4 g / l, K2HPO4 - 0.6 g / l Na2HPO4 - 0.6 g / l. The resulting slurry is sprayed onto the contaminated soil surface, then the soil is digested and crushed. During the cleaning process, the soil layer is thawed periodically once a week. Under natural conditions at 15 - 22 ° C, the soil remediation process takes 2-3 months. The quality of soil remediation is determined by a laboratory analysis of the soil sample based on the residual hydrocarbon concentration in a specialized ecological service laboratory. · Example.
Naftos produktais užteršto dirvožemio valymas aikštelėje. Užteršto dirvožemio kiekis aikštelėje - 100 tonų. Naftos produktų 200 mg/g, bendras teršalų kiekis 20 tonų.On-site remediation of oil-contaminated soil. The amount of contaminated soil at the site is 100 tons. 200 mg / g of petroleum products with a total content of 20 tonnes.
Užterštą dirvožemį sumaišo su trąšomis, turinčiomis N ir P ir/arba kitų priedų, reikalingų minėto mikroorganizmo dauginimuisi. Patikrina mišinio pH, ir jam esant mažesniam už 4, mišinį kalkina. Po to dirvožemį paskleidžia aikštelėje apie 0,5 m storio sluoksniu (esant teršalų didesniam kiekiui, sluoksnis turi būti plonesnis). Paruošia Arthrobacter globiformis MD18 suspensiją: fermentatoriuje Čapeko terpėje išaugina 100 1 kultūrinio skysčio (8-10 g/1 sausos masės). Išaugintą kultūrinį skystį praskiedžia 1χ103 1 mikroorganizmų mitybai reikalingų druskų vandeniniu tirpalu, pavyzdžiui, NaNOs - 4g/l, K2HPO4 - 0,6 g/1, Na2HPCh - 0,6 g/1. Gautą suspensiją išpurškia ant paruošto dirvožemio sluoksnio, kurį po to perkasa. Natūraliomis sąlygomis, esant oro temperatūrai 15 - 22° C, dirvos išsivalymo procesas trunka 2-3 mėnesius. Dirvožemio išsivalymo kokybę nustato specializuotoje ekologinės tarnybos laboratorijoje atliekant laboratorinę dirvos mėginio analizę pagal likutinę angliavandenilių koncentraciją.The contaminated soil is mixed with fertilizers containing N and P and / or other additives necessary for the growth of said microorganism. Checks the pH of the mixture and, at less than 4, limes the mixture. The soil is then spread on the site with a layer approximately 0.5 m thick (in the case of higher levels of contaminants, the layer should be thinner). Prepare a suspension of Arthrobacter globiformis MD18 by culturing 100 l of culture fluid (8-10 g / l dry weight) in a Chapeck fermenter. The culture medium is diluted with 1χ10 3 1 aqueous solution of salts necessary for nutrition of microorganisms, eg NaNOs - 4g / l, K2HPO4 - 0.6 g / l, Na2HPCh - 0.6 g / l. The resulting slurry is sprayed onto the prepared soil layer, which is then reapplied. Under natural conditions at 15 - 22 ° C, the soil remediation process takes 2-3 months. The quality of soil remediation is determined by a laboratory analysis of the soil sample based on the residual hydrocarbon concentration in a specialized ecological service laboratory.
pavyzdys.example.
Naftos produktais užteršto dirvožemio valymas kompostavimo būdu. Vasarą aikštelėje (arba šildomoje patalpoje iki 20°C žiemą) yra 100 tonų užteršto dirvožemio. Naftos produktų 200 mg/g žemės, bendras teršalų kiekis 20 tonų. Į užterštą dirvožemį deda trąšų, turinčių N ir P ir/arba kitų medžiagų, reikalingų minėto mikroorganizmo dauginimuisi ir prideda susmulkintų šiaudų. Po to dirvožemį sumaišo ir patikrina mišinio pH. Esant mišinio pH mažesniam už 4, mišinį kalkina. Po to supila dirvožemio kaupą. Jeigu teršalų daug ir žemės kiekis didelis, kaupe įrengia oro prapūtimo sistemą (aktyvus būdas) arba išvedžioja vamzdelių sistemą natūraliai ventiliacijai paspartinti (pasyvus būdas). Paruošia Arthrobacter globiformis MD18 suspensiją: fermentatoriuje Čapeko terpėje išaugina 100 1 kultūrinio skysčio (8-10 g/1 sausos masės). Išaugintą kultūrinį skystį praskiedžia lxl0J 1 mikroorganizmų mitybai reikalingų druskų vandeniniu tirpalu, pavyzdžiui, NaNCb - 4g/l, K2HPO4 - 0,6 g/1, Na2HPO4 - 0,6 g/1. Gautą suspensiją išpurškia ant paruošto dirvožemio kaupo ir kuo giliau dirvožemį perkasa. Natūraliomis sąlygomis, esant oro temperatūrai 15 - 20° C, dirvožemio išsivalymo procesas trunka 1-2 mėnesius. Dirvožemio išsivalymo kokybė nustatoma specializuotoje ekologinės tarnybos laboratorijoje atliekant laboratorinę dirvožemio mėginio analizę pagal likutinę angliavandenilių koncentraciją.Composting of petroleum-contaminated soil. In summer, the site (or heated room up to 20 ° C in winter) contains 100 tons of contaminated soil. 200 mg / g of petroleum products in the earth, total pollutants 20 tons. Fertilizers containing N and P and / or other substances necessary for the growth of said microorganism are added to the contaminated soil and crushed straw is added. The soil is then mixed and the pH of the mixture is checked. At a pH of less than 4, the mixture is limed. The soil is then poured. If the pollutant is high and the amount of land is high, either install an air purge system (active mode) or drive a pipe system to accelerate natural ventilation (passive mode). Prepare a suspension of Arthrobacter globiformis MD18 by culturing 100 l of culture fluid (8-10 g / l dry weight) in a Chapeck fermenter. Harvested culture liquid was diluted with 1 J lxl0 microorganism nutrition aqueous salt solution such as NaNCb - 4 g / l, K2HPO4 - 0.6 g / 1, Na2HPO 4 to 0.6 g / 1st The resulting slurry is sprayed onto the prepared soil and then digged as deep as possible. Under natural conditions at 15 - 20 ° C, the soil remediation process takes 1-2 months. The soil remediation quality is determined by a laboratory analysis of the soil sample based on the residual hydrocarbon concentration in a specialized ecological service laboratory.
pavyzdys.example.
Naftos produktais užteršto vandens telkinio valymas. Užterštas vandens plotasPurification of an oil-polluted water body. Contaminated area of water
100 m2, teršalų kiekis - 0,2 tonos. Valomą plotą izoliuoja (panaudojant spec. priemones) nuo platesnio užteršto vandens pasiskleidimo. Paruošia Arthrobacter globiformis MD18 suspensiją: fermentatoriuje Čapeko terpėje išaugina 100 1 kultūrinio skysčio (8-10 g/1 sausos masės). Išaugintą kultūrinį skystį praskiedžia lxl03l mikroorganizmų mitybai reikalingų druskų vandeniniu tirpalu, pavyzdžiui, NaNOj 4g/l, K2HPO4 - 0,6 g/1, Na2HPO4 - 0,6 g/1. Paruoštą suspensiją išpurškia ant vandens paviršiaus 5 1/m2 . Natūraliomis sąlygomis, esant oro temperatūrai 15 - 22° C, vandens išsivalymo procesas trunka 2-4 savaites. Vandens išsivalymo kokybė nustatoma specializuotoje ekologinės tarnybos laboratorijoje atliekant laboratorinę vandens mėginio analizę pagal likutinę angliavandenilių koncentraciją100 m 2 , the amount of pollutants - 0.2 tons. The area to be treated is insulated (by special means) from the wider spread of contaminated water. Prepare a suspension of Arthrobacter globiformis MD18 by culturing 100 l of culture fluid (8-10 g / l dry weight) in a Chapeck fermenter. Harvested culture fluid was diluted with 3 lxl0 microbial nutrition aqueous salt solution such as NaNOj 4 g / l, K2HPO4 - 0.6 g / 1, Na2HPO 4 to 0.6 g / 1st The prepared suspension is sprayed on a water surface of 5 1 / m 2 . Under natural conditions, with an air temperature of 15 - 22 ° C, the water purification process takes 2-4 weeks. The quality of the water purification is determined by a laboratory analysis of the water sample based on the residual hydrocarbon concentration in a specialized ecological service laboratory.
Pasiūlytas Arthrobacter globiformis MD18 kamienas palyginus su Azotobacter vinelandii 21 pasižymi tuo, kad greičiau skaido aromatinę naftą bei naftos sunkiąsias frakcijas, geriau įsisavina asfaltenus, aromatinius (antracenas), ciklinius (dekalinas) angliavandenius ir geriau skaido naftą vandenyje. Minėti požymiai leidžia pagreitinti valymo procesą panaudojant aprašytąjį kamieną dirvožemio ar vandens biologiniam valymui nuo naftos ir jos produktų teršalų.The proposed strain of Arthrobacter globiformis MD18 compared to Azotobacter vinelandii 21 is characterized by faster degradation of aromatic oil and heavy oil fractions, better absorption of asphaltenes, aromatic (anthracene), cyclic (decalin) carbohydrates and better degradation of oil in water. These features make it possible to speed up the cleaning process by using the described strain for biological cleaning of soil or water from contaminants of oil and oil products.
IŠRADIMO APIBRĖŽTISDEFINITION OF INVENTION
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3813316A (en) | 1972-06-07 | 1974-05-28 | Gen Electric | Microorganisms having multiple compatible degradative energy-generating plasmids and preparation thereof |
| SU612958A1 (en) | 1976-12-01 | 1978-06-30 | Институт Биохимии И Физиологии Микроорганизмов Ан Ссср | 123 pseudomonas aeruginosa strain carrying oct+, cam+, nah+ plasmides used for biological purification of water from petroleum and petroleum products |
| SU1076446A1 (en) | 1982-07-22 | 1984-02-29 | Западно-Сибирский научно-исследовательский геологоразведочный нефтяной институт | Strain pseudomonas putida 36 for use in purification of water and soil from petroleum and petroleum products |
| LT3111B (en) | 1992-07-15 | 1994-12-27 | Saulius Grigiskis | The oil pollutants decomposing microorganism azotobacter vinelandii 21 |
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1998
- 1998-10-16 LT LT98-146A patent/LT4522B/en not_active IP Right Cessation
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3813316A (en) | 1972-06-07 | 1974-05-28 | Gen Electric | Microorganisms having multiple compatible degradative energy-generating plasmids and preparation thereof |
| SU612958A1 (en) | 1976-12-01 | 1978-06-30 | Институт Биохимии И Физиологии Микроорганизмов Ан Ссср | 123 pseudomonas aeruginosa strain carrying oct+, cam+, nah+ plasmides used for biological purification of water from petroleum and petroleum products |
| SU1076446A1 (en) | 1982-07-22 | 1984-02-29 | Западно-Сибирский научно-исследовательский геологоразведочный нефтяной институт | Strain pseudomonas putida 36 for use in purification of water and soil from petroleum and petroleum products |
| LT3111B (en) | 1992-07-15 | 1994-12-27 | Saulius Grigiskis | The oil pollutants decomposing microorganism azotobacter vinelandii 21 |
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| LT98146A (en) | 1999-05-25 |
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