SP. 168 BEST AVAILABLE COPY

Special Report 168

THE MICROBIOLOGY OFL. TERRESTRIAL CRUDE OIL DEGRADATION

Patrick G. Hunt

April 1972

DEY~

ýA TASZ 4A161 101A9D103

BEST AVAILABLE COPY

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The findings in this report are not to be

construed as an official Department of

the Army position unless so designated

by other authorized documents.

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.. . . .. ..

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Y0CUMUNT CONTROL DATA - R &DI afl.4OOyo10. 60 D 66tM44 w AM i&~dithout *4wotellof Must it's iiftwd W- h vpl [ I0 C40

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C2ROBIOLOGY OF T12RMES]TBIA CRUDE OIL DEGMATAION

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. .DA Task 4A061101oA91D03 uGrie

I~.CI'ME AWPOINT tUols (Aity Ouee wmam Woee£t m~y be =cl.ýMtn.

10.

Approved for public release; distribution unlimited.:

II. SUPPLI.EAdgNTARV N0T96 12. SPOwS*PING MILITAIMV ACTIVIYgy

JU.S. Army Cold Regions Researchand E1ngin,ýering Laboratory

j~ovr 74SS1Ihir ACTE5

It hais been well established in the literature that microorganisms capable ofdiecomposing crude oil ore quite common in terrestrial areas around the world.It hias also been well established that the straight chained hydrocarbons of CIOto C18 length., the kerosene fraction, are the most susceptible to microbial de-gra.datiort1. However., the shorter-chained hydrocarbons., the gasoline fraction.,seems, -1o b-ý degraded more slowly,, and in some instances miay be oicto nricro-

~organ~isms. The ring-structured hydrocarbor~s are also degraded b.1'- -at a consid-illerblyslic-,wer rate than- the straight chained hydrocarbons. The biochemical

Ity ieich hydrocarbons* are degraded are niot unique or few in number.Mr-_ r-cmri to be a substantial void in the literature concerntng bydrocar-.bon

;r'~fl~r> 1- i cold r~egions and the means of optimizing decompositon incld ;~c'1~'~l re-as. It is possible,, bec-cuse of t-he 'Long gene:aticn t-imen oil rd

c2I c) n'- ri r nr wie n s tlr? t oil spills J r' cold Lerre~trial 1-*e1s, ma~y beploaces ý,rhere rncoP.inoei:1aticn. may be beriefici-1t.. llUere

.1" L3.)n ho"L o i-ca ac~tion is li-mit-eci by envi rorumiental 2<ion arcr~nooof microorgani sms.

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1THE MiCKAubIPLOG-1 Q+

TERRESTRIAL CRUDE OIL DEGRADATION

Patrick G. Hunt

S~April 1972

- ~DA TASK, 4ACW3t1IA9V•K03

S~~~~CORPS OF" EN•GINL-FRS, U.S. ,I-.

CO)LD RREGIU RESEARCH AND ENGINEERMHG LABORATORY@SHANOVER, NEW HAMPSHIRE

•" AMPP"VEL.' FOR PU14LIC RULWARE;• DISTRIEU011ON UtNLI"ITE-0.

This paper was prepared by Captaith PatricK G, Hunt, Rescarch Soii Microbl " :.of the Earth Sciences Branch, Research Division, U.S. Army Cold Regions Reseý;..ch -riEngineering Laboratory.

The autho-r wishes to express thanks to Dr. R.P. M-Wrrmann ar d Mv. (A FJck. uUSA CRREL and Dr. R. Benoit of Virginia Polytechnic 1nw:titute foz reviewing, tii inscript.

This iiport is publiahed under DA Task 4A061101691DC3, In-House L-abor.toryIndependent Research.

CONTE~NTS

Introduction ......... . ..... ... 0..... ........ .....

Cntde oil ........................................... ......... ..Crude oil decomposing organisms ......... 2. ....... ..... .

Biodegradability of various crude oil fractions .......... 2Biochemical pathways of hydrocarbon decomposition ... ....... 4Environmental conditions affectiiug crude oil decomposition ....................Literature cited .......... ......... ... ....... ..................Abstract ... ............................................................ 13

ILLUSTRATIONSFigure

1. A schematic of the TCA cycle ......................................2. A schematic CC P-oxidation.........................................,.3. A schematic c( saturated aliphatic dtwcompositlon ........................ 54. A schematic of alkene decomposition ................................ 65. A schematic of aromatic ring cleavage .................................. 66. A schematic of decalin degradation ................................... 67. A s(,ematic of phenanthrene and anthracene degradation to catechol .......8. A schimatic of catechol conversion to TCA cycle constituents ............ 3

MICROBIOLOCY OF TERRESTRIAL CRUDE OIL DEGRADATION

by

P.G. Hunt

INTRODUCTION

The utilization of petroleum for products and services throughout the world has inorfased to

monumental levels and large utilization increases are projected. However. t", benefits that are

realized from the use of petroleum prodi'ets are somewhat negated by the icnidsed probability o0:environmental damage from mishaps iii the handling of petroleum. A part :fly critical, area ofpotential pollution is the transport of massive amounts of both crude anu ocd petrcuim by ship

and pipeline. Notable oil spill disasters such as the Torrey Canyon and Santa Barbara spills havcalready occurred, but considerable progress in prevention and clean-up materials and pr (yedures

has been made (personal observation at the 1971 Conference on Preventicr: and Control fc Oil Spil]h).

However#e4e'nost oil mishaps have been on water most of this rogiress has hri Ia thearea of aquatic spills and relatively little has been done or considered in th57 area of ,:rr-strialspills. Yet numerous petroleum transport systems are terrestrial. For txawoie, the Xr:if•3er

Alyeska pipeline will cross 800 miles of ecologically sensitive terrain in Alaska. Thu te poten-tial for massive ecological damage from terrestrial oil spills exists. Terrestrial oil sp"l" clean-u•is difficvi in any area, but in Alaska, where permafrost soils and slow c"-n. ve,.ta... are pre-valent, the potential problems are magnified immensely., In such a sensitive environm, .-v, physic-,iand mechanical clean-up may cause more ecological damage than the spill itr.,•f. Therefore. afterthe potential water pollution and health hazards have been addressed, c-ne of thte mosat lcicalapproaches for treating a terrestrial oil spill in Alaska is by microbiolocaa teac,. mem Pi;, reportconcerns the topic of microbial decomposition of crude oil in zoils.

CRUDE OIL

Crude oil is composed of numerous organic compounds that originate from direct or indirectbiological action. The components can be classified as paraffins (alkanes), cycloparaff ins (cycio-Salkanes or naphthenes), aromatics, asphalts, and combinations of these (Davis 1967). Otherclassifications of crude oil are also commonly used; Morrison and Boyd (1966) used distillationtemperatures and carbon number to denote certain fractions of crude oil. The composi:tcn andcharacteristics of crude oil vary greatly with source, but compared to the extensive number of possiblecompounds and isomers the variation is relatively small (Meinschein 1959). The aroma tios arenormally iound in lower quantities relative to the paraffins or cycloparaffins, and cyclosxn'aiwc andcyclohexane often constitute large portions of the cycloparaffins. The three primary elements otherthan carbon of the asphaltic, non-hydrocarbon fractions are oxygen, sulfur and nitrogen; in someinstances the asphaltic fraction composes as much as 50% of the crude oil (Davis 1967). The as-phaltic crudes are often more toxic and less bN.deg-adable than other types. However, spillago ofany type of crude oil releasos a large variety o( orgauic compounds into the environi.ient, and zhel,removal with the least health and ecological damage is imperative.

Z ~~MICROBIOL~OGY OF TERRESTE'41f CTZU-Z 1 L DEG P'rt 1 f

CRUOR OIL DECOMPO3SING ORGANIES

Hydrocarbon-utilizing microbes are found from Lhe tropics to the A~ctic an"" 1~"'7" been" is"AU1from Soil, estuaries, ariznal feces, and numerou!ý o' er ioca isn (M,1ge 't d1G''West 1970. Bushnell and Hass 1941). However. ihe -'Gt mpr'tc'"their presence in the high seas to be practically - 3,i"hell ai ' Oxfs

utiti z at ion of hydrocarbon s by t he ge ner a PsezV S' a~ c0b, ,te(1,906), Kaserer (1906), 'St~rmer (190f8), ani I'v -"71 C Qy,

carbon decomposing organisms. lh3 k llGj)Thhy~drc.Tar bons. Streptomyc-es, Nocardia, Myccbacý-cri.Lm, - ,

c~ited as the most commonly isolated hydrocarbon ý`cc<cmpoz, rs -Vi~e IN2number of isolated hydrocarbon decom~pos'trs rep-cwt-0 J, exte-r- 1c:e!>

plating,-and plating is considered to isolaqte only I tc, !107 if tl' rvw 'soil (Casida 1968). Foster (1962), 'or instance, ýý-ec so'mr nr7- C' 11(carbon media but ;,.(ould grow oai the rmore selective md~L" 'carbon souiuce. This serves to illus-;:ate that num-.ý ~~crlanisms ii, t-'-oil at very slow rates. hi:: their isolation arid riýte of oil decompositiou &- ;ý vconditions to include competition with other orgaii M ' or, carýon1 u;ollre-, u!In addition to those organisms that growi on hlydrocar-,s somre organism,4 ,

do not utilize them for growth (Leadbetter and Fo,;!er *;TA 1i.6(..Davi-whe~n the large number of hydrocarbon util~zers and ox'ldi ers iFC (cnsvid red,,hat even though some organisms can d< compos- cortzain l £cttclr-5 of r~~eo;,

>-~others, the decomposition of crude otil in t~et iP c!'.rried Out V rumr' ~ n

)v acv particular speciips at. ai will b*:ýhol--,'''i

Bt1OD~ft A1AB1M~ OF VARWOU CRUDE OIL FRACTIONS

Zobell (1969) stated that more organisms wete capable of decomposing C10 to C18 (kerosenes)than any other fraction. Similar fkiddhg have been iftated by Kazto et al. (1970). This patternof degradation would be intuitively expectcsd since thes compounds are not as volatile as theshorter chained hydrocarbons nor aS sterically complicated as the larger or branched hydrocarbons.Davis (1967) stated that the gaseous hydrocarbonts of low molecular weight, methane through n-butan~e,were oxidized at a rate usually limited by the solub-lity of the gas in the aqueous surroundings ofmicrobial cells. The gaseous phase or 5-10 carbon hydrocarbons was also reported to be utilizedmost readily. However. the liquid phase of these low molecular weight hydrocarbons (paraffins andcycloparaffins) was found to retard microbial growth and in some cases be refactory. Similarly,Davis and Raymond (1961) found that aromnatic hydrocarbons of this size were often toxic to growthof Nocardia, but the toxicity was mitigated by the presence of other degradable hydrocarbonts. Thisstipports the view that the organisms in a natural system such as the soil of an oil spill area wouldbe less affected by toxic components of crude oil than organisms exposed to a medium containinga single toxic component of crude oil as the carbon source. McKenna and Kallio (1965) demonstratedthe variation of biodegradability of hydrocarbons with molecular structure. using gr7owth of speciesof Micrococcus, Pseudomnonas, Mycobacteriumn and Nocardia as their criteria of degradation. Even

.P mhyl group on a straight chained hydrocarb-on redu-ed it- -utilization. and dimethyl or espe-Sgrrvirs grezatlv Teduced m'44frobý.ai tjl>Ziori. Alkanes with icimnal~ phenyl. groiips

- - '~'r~c'~ 'M I!tan es with nhcnyl ':-:,ution alom: the carbon chaln.

A0IcAJoOLOL1Y OF TEiUPESM'1IAL C RUDE;o qL 46HkAV(AT 1 3

Daini~ 11967 P rt~pun"#.0 04". o0taift- of? Cc ft Bol toder both fiold andI laboratory coudittanxiVwvi chactwix" by high Ab34tbAita of iiratA at V1315 rind PAP., hit1j.'ýf wu ge pItr"of aroiwiec sntwflr". tje ulgo found Wth even a~tt 4564A by weisit of c~ttde cM Wiu denmo~l

UWAWa vMnft' I-ii W~~c f[M OIil MidUlf)031Ititlf QXI the~ pmr irfi VI'e ilow dewrAdatkim oftho ~aroILcs w"a* a18 tWdkted by Zabeffl (1960) wbo fow thai polyciekb2-*ebX " VCIKS aam parafYIf fim c t"Ued by bA'twinawm~e readily. thia ~ Wwi iekmn. Thc~oa todifis Luict th'athn highly valastil naM ~toraiht vbauim6 fra.ctimon of i.-:We Ai are likuly to b- m-wae wdfnai ~ai (Ul

ti ~~will Aitlf IpI ofvc u tiid thZ-11 chI ~ifvhly hiaCN-I~d. VnPhIut~t fln d aurmatic tyl'4 coimoffidqi

which am likelly to rentain in an WI1 sral am& fot batended mmiuAA bVOU kwa U~idl;M4I 2A~~

DPNW + IP

MI-31. Co

ItciCCN

HO-0oo

/ ~f- -'DPNH

Thicarboxylic Acid Cycle OýC--COOn(Krebs Cycle)

H,

ft~q H+

GDP C*AMU

Figure 1. A schematic of the TCA cycle (Conn and Stumpf 19636).

4 MICROBIOLOGY OF TF2rRESTRIAL CPj., ? r DEGRAOATIONV

EB!OPHIEMICAh PATHIWAYS OF IIYDROCAR24IX! aE MMP DUIXON

~~ ~ewA) cycle ýnd the beta oxidtv ;'!Twaqy 2Fg I ) wil) t~Otse lx~tersu-termwto h10 the readerý,who is n.ii ok:h fr. 0 rtý-uesv~ ~ b

gr ia jobardbits, othydrocarbo.- are converted to ciaergy, "c's, C92 , n F

Iei6-t~prike~i81cbemistry'of Niese prcsssnr thi tci oV0 U. ~ •

bolites- ie'hddrerssed., These topics aro discussed ab',y b-,; K and Cordes (1%§), Vvbie, 11i 4 w'larid Smtfitf(1,988),". and Conn A, d Stumipf (1IM). The reactions taat are' presented are not intended to

be a&otpiete 1Ws of the hydrocarbon degradztiVe pathways, but they Ir wc, xamples of' how s, vef alfactI60s bf. crude oil 6~n be degraded.,,

R-CH2 -C11 2 -COCH

* ~ 1 ATP(D GAS11

* I R-CH2 -CH 2 -C-CA

CILI~(-C S -C(:A

0

1R-(>-S-CoA C2 01

tram?3

-I Cý 2 02Cli--C--S--CoA 12(

4 0

B-4 -- -S--%CoA

-211

4 * .* *4 .- A

HICRt)DIOLIV~r OF T WE'5TRVAI . 4 MI[I M CbF`PlTIoi'W-

WT-eW I th-~ hinictV 4 hywnu-mifii McKoih*a Aud Kadlio (ji5) liKod ~Novat oxgamIaw

i~~~~~~!dj V '-twmifd k~twen winjo f-heIraY 0h~wxfir twaclaia

ra-vi~tita-i tho tmty thing that mpratami thu Ihyirci'lo fronm typical fatly ar=iti c4 itiv, va0,_1Ar "A

that v t.~ Vw bvmWýVt abWyut by relA-tivoiy o'irmPl lihmi ox mygo tih41 "Md hytcoyktdicwýi AB~b~ziwm vetil Fbr'ý tleb pa . ~ _ d d~ tkt "i!!I-i-(1Aitd! to,1 w'J"My (;(-)A --n C4

i. Akkýion Qv-o~ (.0-A c-au untuc th-3 1,VýA cyrin~~I rsvdu it" to t,, wt UnA i=~ ot w

M(' pwatfic-M11 tin jaeistot with thotr wo-iarvely rapidl ii~eumptiioni~ In uil aspil at#"n.

cKunrat vAe Ka~ul (1(e8) aluso citcnd v~non-_wh Mt lc i u1ucs~ate that Candida. a ynent, CMuldwaft alkmnm by itotinog diols from the double bGmI. but bacteria iureforiti~tally otidiweil the

W"VA gd of Aikehue (tinwart ot iii. 11)61). I-'eudurnon-3 heii Immi nihown to tiograde akikononby ftsm-ation of opozirles " the &xjlo boid. IThese genouel pathways ara preseted in ftgixo 4.Agaiii in. 0-o caane of alkunen the hlnnhomictfl atuop roqaired rft these iomrpouw4. to iintor tho -3m-lI -_

man uzidntivo pvkthwayu aimp ljlo.

Tho cycloparaff ion and artrutictl compoindaw~ havo boon shown to be much less degradablo thanthe alkauuv en aikerien, but thoy are dulraided. Often the degradable side chains rt a ring compouind-_will be deirraded bo(tue thn ring is cleavedl. BiAhi cytcloparaff i and aruntatic rings tond to be tip-

graded vial the fiwmaticm of dudes mid diaciden (E.lliot ot al. 1969), aud dopeadhig upon where the- arcmatic ring breaks a dhIack or uenitaldeiiyde is formned (Fig. 6). Snotu rycitopriraff Inn e-an be con-

'tiwted to tt comirwmo ceIlilar c~mmitutent by relatiVely simlplesap otuc wh as decalin to adipic acidtN p~malin acid (Fig. 6). Adipic. aid nun be idhunted to the 'ICA cycle aui akkcilsic acid by atIoxiygoant~i arnd the. reamual of a terminal aetic) i4rcop as #Aetyl CO-A. Pimulik Aid in involvedin biotin eynthenin in mnone microorganlam O aniIts dlamlne foial in a cell wall conmponet of matnybacteria.

0 c - C- C -

J

G c..H~

:A Iguw 3. A echowaicn~ of styraed 10aI~~n-c decomniilf 1on framMe-Kenna and Ka~ll~o (11GS),

rc

p~gatre 4. A achmLI f~ d-J~~f' c~4 1OKUv 1M~

V HCIOU

indicagv p'oltit of efixyiwe ceIavabe3.

I 1. -'Cool-)

:c e p ico i t. lfr dw-ft~ i'Ad .0 (P ht-njai~ "d K,91110) :;c)

MICROBIOLOGY OF TERI1&STRIAL CRUDE OIL DEGRADATION 7

The moxr complex ring compounds are stertcally more complicated and their degradation ro-quires more complicated enzymes than those required for paraffin. However. even for this group t4compounds a degradative pathway that is relatirely simple can be envisioned. Figure 7 shows Qschevatic f", phemumthrene and anthracene degradatihaxto catechol. Thds degradative scheme iseven more enlightening when it is realized that many organisms readily convert catechol to suc-cinate and acetyl CO-A (T ig. 8) (Lamannaiand Malette 1965).

The asphaltic fractions of iAude oil are degraded by similar pathways, but their degradationinvolves desultation, diamonization or some similar step, that adds another restrictive factor in theirdegradation. Thus it can be seen that the degradation of crude oil components occurs via many verycommon biochemical pathways. However, the reactions can easily be limited by one slow step orenvironmental condition. Seldom are the conditions in soils such that an organism which grows onone of the more complicated components of crude oil will be able to compete favorably with thoseorganisms that grow on relatively simple cotapounds. Therefore, actions that change thb liiltinggrowth factors of hydrocarbon decomposers to optimum levels will probably be the most effectivemeans of increasing the rate of microbial degradation of crude oil in spill areas.

knthroc ens- I

Phen~onthians 8

HO HO H 0 OH

3,4-D1hydro-3,4-dl .!ydroxy- 1, 2-D! hydro-wI,2-dlhydroxy-Phenonthremb anithracene/

OH// ¢/•~'NCOOH i•O

0 H

W,,5

H~~G I' 2H CU" GOA

ALL IIQwe na-d al' rawrehai 'ovnfr-c4tq ro rC-A cyck 'ittofi._

153uff is" LWV rlWi 1W4, 'Igjnýf thi'' IyVilCU-MI fUNWly MIN 1 r-~mA otQit sh a I"Ugt teMrnarrIz WAD. BUYl ti g irlllpru-

(~~p' .-- q'i ~~'fl&M Js~l, keit-ti-si, t- . ''h.-i s;ial'st-3 C, crudn oil ma-y quickly trstvo 1sP15

s>' dJirerunt141v~r-II'1 5'~tth L'IIinWWY'- oVodlvd -' (11 V'A ~ fv;e-rr"e tw eovsrtcd "An a cut s~vUt.s

''½' '' k~i'* tnc' t''•UtsZ boIo col'a ului Ns'wis? hym=-ntI fms~ipi . )¶s~tw~itty 4 :tailm Iob, N" ni

:-st vok ;, OIs lb " kt''"Ahw Lun vsjdt-Itis' of Luu-LwanLha d AintAttk thoii,'I-&c usmeiplsi w0 - A5 "bo W'-"',m Ai'lhi~to U" mi':sl (AbrYsuln 1331y, 'Oiion, a 1Limo jýpnpUI•.l' list mp'4

5

R. jI PP2tf vcs injsapI r din Yk it muat Lu abluo 1Kn i ms"m vitifiduIu pztnn~hI.

- -~- ~i'-'f~ *titA!~U''J t~y OZ QI'ir'b MIAr Ml IM Ali -JA UQJi'il 1nis-h, a9 isfrn K i sill 011nPi , lit- M I sslrdlW~ aLL. 3ý A' 9) rinlLI-.atli ito oitozs lW-s mvalibsilly ofa! om unr Ioniiuu tsP't, olarolilallv Isbntlon.:'ss

I'. l~i2; 5 V i'2.fliithi$4. ll i-x-'3i1y 'Wviiflali-s ltki WIVsp!ulstu (A 1.srYJ4As-ris1'ss .Y-

-'--'= a.- bit i5tsUmjV'tAt'3-1 wdrh thl utllirv A! thei 4J-IiAIPUTIpsllIEt/ kioth. 11d- tv/u tygi

o! no Wi.s1~4.ilk likoiy to) jwi !:rdse uni sim"u- in a *Oyel II (a,1tltnI w041 o-nostre'; kf

tb-ý [--Mn' 5.AM ouS Waya d 6Mt''-Aai' ds-• Irmo" 4 CIAO oil Mg'o~a-ssh It.ev sulnl1 ic tintil1-

'53 'b iSlit W1 " 5914 t n ~nfIlJ1 l-a, si nsd 24h asiý 1- ' ugluiŽtitv' 'is'Its-'i

- -- -AI V of. M45 1''W loa1 z"0

f- t' ll sis wnV cmin-Mi Ms-sr!0 .' ,- liI' 4'~

I ~m f.~-'5W4L vitSjI~.;{~ lVW~ "t L? M I''!''ap-,a 01,, iý.... i si4 t0 !''boI-1s'

Q24 o4.914 11 4~ 'o.;-' -1'' ~~'.

-~~~~i FJF A..-i -

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I htitt ~ Ot. IM, tou4n -Mi.dlo - at-i t,4

W~'oid i:L4 . IJno UMi Žfio vrxi'Lj by a.U ralin it *

""1:1 O itO i of -9Y M.h t lIpA ini II{1( W aji" wht Usiter zllhill[! iOfi UAYiti p ul. VJjit It'r1:6m8ik OL ~i uIg d"E

cgupv~~1)fi in ndditwonn wik y Iv:uin. nXIi-W t4 piyon"Mpim' (.q191 I thuczi dthatsa [me A-

fwilba. In lf tvypit9,x to ~iutiUn ttWrOCithitl aM-"1U4Ati wundiulSM bti wa x die~yslow 00imrn ULh e Uiiii

IMN~'rw ant 01-m~n W ay is, reqIre 0 MrNhwuea n uA.

1 iwi L~g twtin,.~ Aluwp atv d -,loc inmx si atiofol pllrntagbecuuilrnas

mu' t~~ oxyg 1 ditu (1 .:emigh W ftm"Q raply thmwlilih). oils aph- j 7itb~y whyr~i oerdng niws-rs-

Itx tth iitrntulilnhi- have-i) lower ra~n Ilu,; o c'tvivru- knlhtuM a-I~t~prtrtsi ~~kohi's wtAit- 'in a-uit ioni work byiiiu M eim", iuri" aI kwe li-jrtia (1wc71) 'irim aeti!to ha i-t asiný m

wajtp s .,t~iC~ii witil ,A- munz. str-iam Itruph Ava guv;r "r~i SNMM oil 'r~~ted i tMti-i hit UN--641 pC)'C ti

file.wIIt;ia thi typ enhW smion~u~ Iim"obIi ho~ntetiuimi' woul Wi~"~ ui-ieuly islow, an w-tthe-~u p"svlty 21

Ccru-vye iv water~n' P01 tttitltov and i bat;h 'insulttisiii;.uLu t i. low. Itucnift-M'd~t "Man -o ilt %p3t ildifn

T-ompmat~ ureo hit- v:, moor Alul-tIe Wr?.t rm 1ute Wuhd chmiwv I' mH-tt bntgis' h trmuii' tiiy am.1 iuho low

html:;ilitittlastd VIuo t, mti mini t dlytc Man r un We cooII~ of1' muobi;l sp'uilt. reovr byi mitt rubtdianilumi'At itt[ irnetIl_, ,ledI. tite2 III MMl ii fIIlm.tin nuMauini irUWWINruho Lie OWW"0pii "Wi W it AWil- tintm'iI I-

avt muI as. 25i toit30 thld igu (Lumallunit'' 'v d'l'lltu m 1% l6V. Tl iIIv; probabl why (,it degrading j2 t -lm hlu-

ot -gai as- hav bwan ttmlrrthmrv to hwtisi OW late. oft~ ).ritruo f~~c. r"0-t!" Pnr

nio-1iui it;WW Oraktl l il !;u hid -, at'r. 'n 'ill 2iho Noah ":-!ka I t i:tofta tykc ýueovt!m iditl fI W

a i lenin It'1 104 CITU r .li'l'Illttt i ti uw titini:';' ~ ~ n u nmnouw yimiin t

10 tMICROBIOLOGY' OF TERRESTRIAL CRUDE OIL DEGRADATION

LITRATURE CrIED

Alexander, M. (1961) introduction to so&i microbiology. New 'Vork: John Wiley and Sons, Inc.,

472 p.

,Brown,, J. "nd O.C. West (1970) Tundra Biorne research "In Ala:.ia: The structu¢a and function

cold-dominated ecosystems. U.S. Tundra Bluome. Lwornatioa! Dloiogical Pr•,Zm 1,0 p.

t ?Availible from Tundra Bluors Center, Univ. of Aatz.)

-"Bushnell, L.D. and H.F. Hass (1941) The utilization of certain hydrocaxbons by iroo•nisr.:,

Journal of Bacteriology. vol. 4 1, p. 653-672.

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~T.,

... .. .. , .*.h; '~'r"v r~ ~.'' ,~51

-7 *,

MICROBIOWAGY OF TERRESTRIAL CRUDE 011, DECP VAATION 'A

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