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PROCEDURES

The phenolics biodegradation data froe the 1982 hndhrlling prograa val reviewed for 'the purposes of (1) evduating the effec­tiveness of biological treablent and (2) recotl8endating treatment procedure• for the 1983 prOC)raa. After co.pletion of the 1982 prograa, bench scale phenolics biodegradation studiaa vera b89un in aoil bo•a• cont.lning aoil fro. tha traataant alta and t n atattc flaak culturaa uatng a phenol-baaal aalta aediua. In both traataant •atricaa, tha affect of coeubetrata addition ta belOCJ evaluated.

Polloving taraination of the 1912 landfaratng protraa on October 21, phenolica contaainated aoil vaa collected fr• quad­rant one. '11\a 1011 waa tranaported to OMM'a Plndlay, Ohio labora­tory and aubdivided into IOU bo••• (21• • 21•• ,., for biodegra­dation atudiaar aoil depth ln ..ch boa vaa I lnchaa.

each ::ry:"!:."~!:.09:; :~..::::::o-!,;:!:.~= :1 := ~:i!~"p11 •• edjuoted to within on optlaol ro- fo• bac:toflol ,.t.th ••lot

t!:':';: =::!:t!~2.!.10t) ~!trt!:~0:=!t:::i::;c:.:c~•l:!:f; wre Miatai... witltia ae opt!._ l'IDI• fol' Mctel'ial ll'owt.h. aou -••t•r• coeteet .....iatai... wltltla ,. ta 10 parceat of tloe •htuo 1oo1•1.. -ltr, - r.-ur tilled, - •• th.. Miat•l... Ia a Milt, looM eo•Utt•.

lloo •,.•'-tel -It• to• t1oe •11 looo ot..r wo oo fol-1-•

Cj!!Pttrott M!!!!J!

11­·-t btrect81_... + ,..at ntl'act .. IMitiM

Ja ...ltl• to .-tterl.. tM iMqMic • aftlrf t ceeeeatrationa, .. 1...1, _.Mil •t•t•l'• coet-t, •11 Mctedal ,.,.latiNa_. tetal .....Uca coaceatl'atloea were ..temi... laltlallr aadl par i­..lcallr tlilerMfter fer eecta of tM aoU ....... .,.ltloaa of tltM:OM aM./er J"Hit ••tract wre parlodlcallr .... to the dealt· aated eon .....

Ia ...itloa to the eoU lao• atody, tiM effect of coaubatrete addition on phenol d19radetion Ia MiDI deta..ine4 Ia 1 baaal aalU ..... uiat eoU tnoc:.ala fr• a •teUlo Mil -ple. With r19ard to eapert..ntel delltn, co•ubatrate addition• are ai•ll•r to tho•• for the aoll boaee ..

Prior to initietlon of the 1011 boa ltudy, an ••tract fro. • 5 grM 1011 1•pla in 10 ailliliter1 ••thenol VII analyae4 for phenolic content by OC/Ma procedurea. The purpoae of the analyala val to identify the principal phenolica r•aining in the aoil fol ­loviOCJ the 1912 landfaralng prOCJraa.

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RESULTS AND DISCUSSION

The sample means and 95t confidence limits for th.e colori­metr ically quantified total phenolics and the microbial aerobic heterotroph concentrations for soil umples collected from the treatment site during the 70 day 1983 landfarming program are pre­

\ aented in Figure 1. Results demonstrate an exponential decrease

\ i~.!~:.~~·~~!~~=m~on!~~~tl~n t~~ !~!~1 f~~!~~l~Y.!"!i 1~~!~~.~~.~~· qlucoae to the treatment site significantly increased the pheno­lics degradation rate. The cosubatrate enrichment technique was inithted on day U and was utilized through the remainder of the 1982 hndhrming program.

Dur ing the fh'st 13 days of t r eatment, total phenolics con­tent of the s oil WiU reduced from a mean value of 51 6 t o 247 •Uligra•a tohl phenolic• per kil ogram aoil . Using the linear regreaaion line-of-beat-fit through the data, a daily removal rate of 21 -.g total phenolic:a per kilogram aoil waa achieved. That rate of r•oval h equivalant to 68 pounds of phenolic:a removed fr011 the 1700 cubic: yarda of soil per day or a total of 880 pounds of phenolic:a rMoYed fro. the aite in the firat 13 days of treat­

"") ••nt ("'J totol ~enollco/kg ooll x3 lkg/ lOOOg x lg~lOO~IIl<J x ..._) 70 • 34 lbl 1011/ft IOU X 1700 yda IOil X 24 ft / yd • lba

total phenolic:a). ·

roUowinv detradation of the •ore easily metabolized pheno­lics, the eoaubstrate enrichment technique (1,3 , 8,10) usin9 glu­coae as c:oaubatrate (2,4-7,9), waa uaed to inc:reaae the biodegra­dation rate of the •ore recalcitrant phenolica that remained. Ca.p.ring the alopee of the linear regreuion linee-of-tt.at-fit throuth the data, phenolica degradation rate waa 5. 74 timea treater for daya 41 th r ough 70 than that achieved for daya 13 throuth 41; i.e., total phenolica reduction in t he aoil following coaubatrate addition waa 69 pounda tiJiova1 per week. Following coaubatrate llddition, •e•n (geometric) microbial aerobic: hetero­troph population• in the aoil increaaed by a factor of 100; i.e., the •ic:robilll popul11tion waa 100 timea greater on day U than on day 41 and that increaald population denai ty remained throughoutthe r ..ainder of the 1982 landfarming program.

Prior to initiation of the bench acale atudiea in the Findlay, Ohio lllboratory, a aample of the aoil to be used in those studiea waa analyzed by GC/ MS procedures to identify the principal phenolica remaining in the soU. o.s. EPA priority pollutant phe­nolica, which were initially found, were no longer detected in the treatment aite aoil. 'l'he principal phenola found remaining in the oil were m-.!!!!-butylphenol and p-1, 1, 3, 3-tetramethy1buty1phenol.

60

0

500

40

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200

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40

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, 1/

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. T

he Slllp

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•its fo

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e colo

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uan

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olics

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Preliminary data from the bench scale studies supports con­tinued use of the cosubstrate enrichment technique as a means to significantly increase phenolics biodegradation rate in the soil environment. At the start of the landfarming program, colorimet­ric total phenolics analysis indicated 1670 pounds of phenols con­taminated the 1700 cubic yards of soil. That was mean value based on the anal yah of two seta of four compost te samples, one com­posite per quadrant, and each composite was baaed on the collec­tion of eight randomly collected soil samples per quadrant. By the close of the 1982 landfarming program, 1120 pounds of phenols had been removed from the treatment ai te, leaving only 550 pounds remaining. Removal of an additional 220 pounds will leave the treatment site with a total phenolics concentration of 100 milli­gn.ma per kilogram soil which is the goal of the treatment program.

CONCLUSIONS AND RECOHHENOATIONS

Based on (1) the total pounds of phenolics removal from the tteatment lite duri ng the 1982 landfarming program, (2) the effec­tive rate of phenolics removal from the soil at the close of the 1982 pro<Jrarn, and (3) the preliminaty resultl of the bench scale studies, it is recommended to initiate the 1983 laridfarming pro­gram, as soon as possible, in order to complete the phenolics cleanup in the contaminated soil at the site during ideal weather cC?nditions for microbial biodeqradat"ion.

}

LITERATURE CITED

1Beam, H.W. and J.J. Perry, "Microbial Degradation of Cycloparaf­finic Hydrocarbons Via co-Metabolism and commensalism," J. ~ Microbial., 1974, 82:163-169. ­

2cox, D.P. and A.L. Williams, "Biological Procesa for Converting Naphthalene to cis-1, 2-Dihydroxy-1, 2-Dihydronaphthalene,"!221:.. Environ. MICrobial., 1980, 39 (2): 320-326.

3Horvath, R.S., "Cometabol ism of the Herbicide, 2, 3, 6-Tr ichloro­benzoate by Natural Microbial Populations," Bull. Environ •

..,.....-.., ~~· 1972, 7(5):273-276. ----­

(~orvath, R.S., "Microbial co-Metabolism and the Degradation of Organic Compounds i n Nature," Bacteriol. Rev., 1972, 36(2) : 146-155.

5Horvath, R.S., " Enhancement of co-Metabolism of Chlorobe nzoatea by the Co-Substrate Enrichment Technique, " ~ Microbial . , 1973, 25 (6) :961-963.

6Horvath, R.S. and a.w . Koft, "Degradation of Aklyl Benzene ~ Sulfonate by Pseudomonas Species," ~ Microbial., 1972,

· .~ 23 (2) :407-414.

Horvath R.S. and P. Flathman, "Co-Metabolism of Fluorobenzoatea by Natural Microbial Populations," !E!2h ~Microbial., 1976, 31(6) :889-891.

1Rubin, H.!. and M. Alexander, "Effect of Nutrients on the Rates of Mineralization of Trace Concentrations of Phenol and p­Nitrophenol," Environ . !!:..!...:. Techno!., 1983, 17:104-107.

9sethunathan , N. and 11. D. Pathak, "Development of a Oiazinon­De4Jrad ing Bacterium in Paddy Water after Repeated Applica­tion• of Diazinon," £!.!!.:_:!.:,Microbial., 1971, 17:699-702 .

10Tabak, H.H., S.A . Quave, C.I . Maahni, and !.E. Barth, "Biodegra­dability Studies with Organic Priority Pollutant Compounds," ~!!!!!.!. !.21!.:. ~ !!:!!.:.· 1981, 53(10):1503-1518.

11united States Environmental Protection Agency, Methode for Chem­ical rnalyiis of Water and Wastes, EPA-600-4~,£nviron­iii'i'nta Hon torTng~Support La oratory, U.S. EPA, Cincinnati, OH, 1979 .