EPA/540/R-96/505
                                September 1997
         SITE  Emerging Technology Report
Innovative Methods for Bioslurry Treatment
                                           by:
                                   Kandi Brown
                                IT Corporation
                    San Bernardino, California 92408
           EPA Assistance Agreement CR821186-01-0
                           IT Project No. 408250
                                 Project Officer:
                                 Brunilda Davila
     National Risk Management Research Laboratory
  National Risk Management Research Laboratory
              Office of Research and Development
            U.S. Environmental Protection Agency
                          Cincinnati, Ohio 45268

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                                      TECHNICAL REPORT DATA
                              (Plane read Instructions on the mmt before completing]
1, REPORT NO.
  EPA/S4Q/R-96/5Q5
              3. RECIPIENT'S ACCESSION NO.
                   PB97-176820
4, TITLE ANOSUBTITUi
              S, REFQRT DATE  , ft-_
                   September 1997
  SITE Emerging Technology Report:
  Innovative Methods for Bioslurry Treatment
              6, re,BF0«t«»NO ORGANIZATION CODE
7, AUTMOR(S)
  Kandi Brown
              S. PERFORMING ORGANIZATION REPORT NO,
9, PERFORMING ORGANIZATION NAMS AND AODWiSS
  IT Corporation
  1425 South Victoria, Suite A
  San Bernadino, CA 92408-2923
              1O. PROGRAM ELEMENT NO.
                   TD1Y1A
              11. CONTRACT /GRANT NO,
                   CR821186-01-01
!~2. SPONSORING AGENCY NAME AND AOOBiSS
  National Risk Management Research Laboratory
  Office of Research and Development
  U.S. Environmental Protection Agency
  Cincinnati^OH 45268	_	_	_

                                        COVERED
              14, SPONSORING AGENCY CODE
                   EPA/600/14
15, SUPPLEMENTARY NOTES
  Project Officer: Brunilda Davila (513)569-7849
18. ABSTRACT
  IT Corporation conducted a pilot-scale testing of biological and chemical oxidation of slurry-phase polycyclic
  aromatic hydrocarbons (PAHs). The 7-month demonstration illustrated potential effectiveness of combined
  biological and chemical oxidation for the treatment of PAH-contaminated soils.  The 30 percent CPAH
  destruction goal was achieved with CPAH transformation ranging up to 84 percent.  A system hydraulic
  residence time (HRT) of approximately 37 days increased system  performance.  Due to  the increased
  transformation of PAH in reactor I (Rl) and reactor 2 (R2) during optimal performance, transformation
  in reactor 3 (R3) were significantly decreased.  This result may indicate that R3 is not required for effective
  treatment.  It should be noted that greater than 80 percent CPAH removal was achieved during the last 2
  weeks of operation following modifications to the treatment process made during the previous weeks. These
  results are reflective of the effectiveness of the treatment system following achievement of'steady
  operation.  The modification to the treatment system included increasing the system HRT from 13.5 to 37
  days. This change resulted in a HRT in Rl equal to the previous system HRT. As a result, PAH and CPAH
  removal increased in Rl and 12, with a decreasing performance in R3. Overall, operation of Rl and 12 only
  was adequate for effective treatment following in 'increase in HRT, Continued investigation under the process
  set points maintained during the final month of system operation is recommended.
17.
                                   KEY WORDS ANO DOCUMENT ANALYSIS
                    DESCRIPTORS
   PAHs, CPAHs, biodegradation, Fenton's,
   chemical, oxidation, bioslurry reactors
                                                   •.IDENTIFIERS/OPEN gNOED TERMS
                             c.  COSATI Field/Croup
18, DISTRIBUTION STATEMENT

   Release to Public
                                                   IS. SECURITY CLASS (This Report)
                                                                                 1. NO, OP ?AG
      Unclassified
20. SECURITY CLASS (This pagtl

      Unclassified	
                              22. PRICE
 6PA Form 2220-1 (R»*. 4-77)    previous COITION is OBSOLETE

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 Notice
The U.S. Environmental Protection Agency through its Office of Research and Development
partially funded and collaborated in the research described here under assistance agreement
CR821186-01-0 to IT Corporation.  It has been subjected to the Agency's peer and administrative
review and has been approved for publication as an EPA document.  Mention of trade names or
commercial products does not constitute endorsement or recommendation for use.

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Foreword	

 The U.S. Environmental Protection Agency is charged by Congress with protecting the Nation's
 land, air, and water resources. Under a mandate of national environmental laws, the Agency
 strives to formulate and implement actions leading to a compatible balance between human
 activities and the ability  of natural systems to support and nurture life. To meet this mandate,
 EPA's research program is providing data and technical support for solving environmental
 problems today and building a science knowledge base necessary to manage our ecological
 resources wisely, understand how pollutants affect  our health, and prevent or reduce
 environmental risks in  the future.

 The National Risk Management Research Laboratory is the Agency's center for investigation of
 technological and management approaches for reducing risks from threats to human health and
 the environment.  The focus of the Laboratory's research program is on methods for the
 prevention and control of pollution to air, land, water and subsurface resources; protection of
 water quality in public water systems; remediation  of contaminated sites and ground water; and
 prevention and control of indoor air pollution. The goal of this research effort is to catalyze
 development and implementation of innovative, cost-effective environmental technologies;
 develop scientific and engineering information needed by EPA to support regulatory and policy
 decisions; and provide technical support and information transfer to ensure effective
 implementation of environmental  regulations  and strategies.

 This publication has been produced as part of the Laboratory's strategic long-term research plan.
 It is published and made available by EPA's Office of Research and Development to assist the
 user  community and to link researchers with their clients.
 E. Timothy Oppelt, Director
 National Risk Management Research Laboratory
                                             m

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A bstract	

 IT Corporation (IT), Knoxville, Tennessee, in collaboration with U.S. Environmental protection
 Agency (EPA), investigated the feasibility of combined biological and chemical oxidation of
 polycyclic aromatic hydrocarbons (PAH).  Bioslurry treatment of PAH-contaminated soils was
 demonstrated under the Superfund Innovative Technology Evaluation - Emerging Technology
 Program (SITE ETP) as an extension of research previously funded by IT Corporation (IT)
 (Brown and Sanseverino 1993) and additional investigations supported by the U.S. EPA (Davila
 et al. 1994). All testing was initiated in September, 1994.

 During the demonstration,  IT operated two 60-liter (L) TEKNO Associates bioslurry reactors
 (Salt Lake City, Utah) and a 10-L reactor in series under semicontinuous, plug-flow mode for a 7-
 month period.  The first 60-L reactor received fresh feed daily  and supplements of salicylate and
 succinate to  enhance PAH biodegradation.

 Slurry from the first reactor was fed to the second 10-L reactor, where Fenton's reagent
 (Fe+++H2O2)was added to accelerate chemical oxidation of 4 to 6-ring PAHs.  The third reactor
 in series was used to biologically  oxidize contaminants remaining following addition of Fenton's
 reagent. This reactor received no additions of salicylate and succinate and was aerated, nutrient
 amended, and pH adjusted only.

 During  operation, the reactor system demonstrated total PAH  and carcinogenic PAH (CPAH)
 transformation up to 95 and 84 percent,  respectively.

 This report was submitted in fulfillment  of assistance agreement CR821186-01-0 by IT
 Corporation under the partial sponsorship of the United States Environmental Protection Agency.
 This report covers a period from  September 1994 to April 1995, and work was completed as of
 December 1995.
                                             IV

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Table of Contents.
Foreword  	  iii
Abstract	  iv
List of Tables	yii
List of Figures	  viii
List of Acronyms	  ix
Acknowledgment	xi
1.0    Introduction	1
   1.1      Site Description  	1
   1.2 Soil Characterization  	  1
   1.3      Waste Stream Description	  1
   1.4     Remedial Technology Description	2
2.0    Conclusions and Recommendations	4
   2.1      Conclusions   	 4
   2.2     Recommendations	4
3.0    Treatability Study Approach	6
   3.1      Test Objectives 	 6
   3.2     Experimental Design and Procedures	6
   3.3      Equipment and Materials	  7
           3.3.1      Soil Collection and Preparation	7
           3.3.2      Reactor Description and Operation 	9
   3.4     Sampling and Analysis	  10
           3.4.1      Physical  Analyses  	  11
           3.4.2      PAH Analyses	11
           3.4.3      Microbial Enumerations	  12
           3.4.4       14C Mineralization Assays  	  12
           3.4.5      Toxicity Screening	  12
           3.4.6      Chemicals  	  12
   3.5     Data Management	  13
   3.6     Deviations from the Test Plan and QAPP  	  13
4.0    Results and Discussion	  14
   4.1      Data Analysis and Interpretation 	  14
           4.1.1      Reactor Operation	  14
           4.1.2      Physical  Analyses  	  14
               4.1.2.1  pH  	  ,	  14
               4.1.2.2  Dissolved Oxygen	  15
               4.1.2.3  Reactor Solids	  15
               4.1.2.4 Nutrients	  16
               4.1.2.5  Microbial Enumerations	  16
               4.1.2.6  Total Recoverable Petroleum Hydrocarbons	  17
               4.1.2.7  Total Organic  Carbon Analyses	  17

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Table of Contents (continued).
               4.1.2.8   PAH and Mass Balance	  18
               4.1.2.9   14C-Mineralization Assays  	20
               4.1.2.10 Toxicity Screening	21
   4.2      Quality Assurance/Quality  Control	21
   4.3      Costs/Schedule for Performing the Treatability Study	21
   4.4      Key Contacts	   	21
5.0   References	22
Tables
Figures

Appendices  -  (Complete copies of all listed appendices  may be received by  contacting the U.S.  EPA
Project Manger, Bruni Davila [513-569-7849].)

Appendix A    Geotechnical Reports
Appendix B    Site Data
Appendix C    PAH Data and Mass Balance Spreadsheets
                                           VI

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List of Tables	

Table               Title

   1-1                Physical/Chemical Properties of CPAH Constituents
   1-2                General Influent Feed Characteristics for Bioslurry Treatment
   3-1                Initial Operational  Setpoints
   3-2                Sampling and Analytical Schedule for Rl, R2, and R3 Slurry Phase
   3-3                Summary of Analytical Methods
   3-4                Summary of Chemicals
   3-5                List of Technical Changes
   3-6                List of Nonconformances
   4-1                Summary of Reactor Upsets and Operational Modifications
   4-2                Summary of Total Recoverable Petroleum Hydrocarbons (TRPH), PAH,
                     and CPAH for March 16 Samples
   4-3                PAH Removal Efficiencies (Percent)
   4-4                CPAH Removal Efficiencies (Percent)
   4-5                PAH Mass Removal (Grams/Day)
   4-6                CPAH Mass Removal (Grams/Day)
   4-7                16-Hour Toxicity Determination Using Tetrahymena
                                          vn

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List of Figures^
   3-1
   3-2
   4-1
   4-2
   4-3
   4-4
   4-5
   4-6
   4-7
   4-8
   4-9
   4-10
   4-11
   4-12
   4-13
   4-14
   4-15
   4-16
   4-17
   4-18
   4-19
   4-20
 Title

Process Flow Diagram
Eimco Biolift™ Reactor
Reactor pH Data
Dissolved Oxygen Concentrations
Reactor Total Solids
Reactor Volatile Solids
Ratio of Volatile Solids to Total Solids
 Solids  Distribution in Reactor 1
 Solids  Distribution in Reactor 3
Reactor Ammonia Concentrations
Reactor Phosphate Concentrations
Reactor Total Heterotrophic Bacterial Populations
 Solid Phase Total Carbon
Aqueous Phase Total Organic Carbon
Percent Reduction - Rl
Percent Reduction - R2
Percent Reduction - R3
Percent Reduction - Overall
Fluorene  Concentrations
Benzo(a)Pyrene Concentrations
PAH Concentrations
 CPAH Concentrations
                                    Vlll

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List of Acronyms,
                 Biotechnology Applications Center
                 Benzo(a)pyrene
                 colony forming units per milliliter
                 Chemical  Hygiene plan
                 carcinogenic polycyclic aromatic hydrocarbons
                 dichloromethane
                 diatomaceous earth
                 U.S. Environmental Protection Agency
                 Emerging Technologies Program
                 gas chromatography/mass spectrography
                 sulfuric acid
                 high performance liquid chromatography
                 hydraulic retention time
                 IT Corporation
                 liter
                 lethal dose
                 molar
                 milligrams per liter
                 milligrams per kilogram
                 manufactured gas plant
                 milliliters
                 millimolar
                 millimeter
                 Normal
                 nanometer
                 polycyclic aromatic  hydrocarbons
                 pentachlorophenol
                 process flow diagram
                 quality assurance project plan
                 Reactor  1
                 Reactor 2
                 Reactor 3
                                   IX

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List of Acronyms  (continued).
 rpm          revolutions per minute
 SITE         Superfundlnnovative Technology Evaluation
 SOP          standard operating procedure
 TC           total carbon
 TOC          total organic carbon
 TRPH        total recoverable petroleum hydrocarbon
 TS           total solids
 TSDF        treatment, storage, and disposal facility
 UV           ultraviolet
 VS           volatile solids

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A cknowledgment

 IT and U.S. EPA would like to acknowledge the contribution of Gunter Brox, TEKNO
 Associates, during the execution of this work.
                                      XI

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1.0 Introduction
IT Corporation (IT) was contracted by the U.S. Environmental Protection Agency (EPA)
under a cost sharing contract (CR821186-01-01) in October 1993 to conduct pilot-scale testing
of the biological and chemical oxidation of slurry-phase poly cyclic aromatic hydrocarbons
(PAH).  Bioslurry treatment of PAH-impacted soils was demonstrated under the Superfund
Innovative  Technologies Evaluation -  Emerging Technologies Program (SITE ETP) as an
extension of research previously funded by IT (Brown and Sanseverino 1993) and additional
investigations supported by the EPA (Davila et al. 1994).

All testing  was initiated in September, 1994. Testing was conducted by IT personnel at IT's
Biotechnology Applications Center (BAG) located in Knoxville, Tennessee.

1.1    Site  Description
Among the types of contaminants present in Superfund soils, complex PAHs constitute the
more challenging class to treat.  Sites that contain PAH contamination include manufactured
gas plants (MGP), wood-treating facilities, petrochemical facilities, and coke plants.  Soils
employed during this investigation were collected from a wood-treating facility located in
Arkansas.

1.2    Soil Characterization
All soil collection and screening  activities were conducted by IT personnel, with supervision
by the wood-treating site health and safety officer during the week of September 12, 1994.
For a complete review of soil screening activities see Section 3.3.1.

1.3     Waste Stream Description
PAH and carcinogenic PAH (CPAH)-impacted  soils, primarily sand (30 percent) and clay (70
percent), were wet-sieved on site through a 30 mesh screen and submitted to  IT's  BAG, for
testing. All geotechnical analyses are  presented in Appendix A. Oversized material was
disposed on site. Blended slurry PAH and CPAH concentrations ranged up to 6,120 and 434
milligrams per kilogram (mg/kg), respectively. Wet sieving the soils increased the uniformity
of the slurry,  thereby, reducing the potential for sampling variability.

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 1.4    Remedial Technology Description
PAHs are characterized by high organic partition coefficients, low aqueous solubility, and low
vapor pressures (Table 1-1). These characteristics result in the highly sorptive nature of PAHs
and their subsequent limited availability to microbial populations.  IT's past experience with
PAH-contaminated soils indicated that contaminant desorption from soil is the rate limiting
factor in bioremediation. Manipulation of parameters such as pH, agitation, and temperature,
as well  as the addition of surfactants or solvents, can be used to enhance the rate of desorption
and, thereby, increase in the rate of biodegradation.

The optimum method of manipulating these parameters is in bioslurry reactors. Bioslurry
reactors can provide rapid biodegradation of contaminants due to enhanced mass transfer rates
and increased contaminant to microorganism contact.  These units are capable of treating high
concentrations of organic contaminants in soils and sludges, with demonstrated biodegradation
of selected contaminant concentrations ranging from 2,500 to 250,000 mg/kg. In general, the
percent  removal of PAH in  these systems ranges from 70 to 95 percent, with 30 to 80 percent
reduction of the CPAH fraction (EPA, 1990).

Bioslurry  reactors can aerobically biodegrade aqueous slurries created through the mixing of
soils or sludges  with water.  Maximum contaminant reduction is accomplished in bioslurry
reactors primarily through proper feed preparation.  Preparation of the influent waste stream
should produce the general characteristics presented in Table 1-2.

The most common mode of bioslurry treatment is batch; however, continuous-flow operation
can be achieved.  Aeration is provided through floating  or submerged aerators or compressors
and spargers.  Mixing may be achieved through aeration alone or in conjunction with
mechanical mixers. Nutrient addition and pH  adjustment are accomplished through metered
chemical addition to the reactor. Following aeration, the treated slurry is dewatered via
standard dewatering equipment, such as clarifiers or filter presses.

The residual streams created during bioslurry treatment include treated solids, process water,
and possible air  emissions.   The process water collected during the solids/liquid  separation
phase is usually recycled for influent waste stream slurrying or discharged under permit.  Air
emissions may be controlled through air pollution control devices.

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Full-scale commercial bioslurry units require approximately 0.5 to 1 acre per million gallons
of reactor volume (EPA, 1990).  Reactor size is determined based on the hydraulic retention
time (HRT) required for treatment. Retention times are established based on the
biodegradability of the waste, level of treatment required, influent  contaminant concentration,
and physical/chemical nature of the waste.

Major issues of concern during bioslurry treatment system design include reducing system
HRT  and increasing the rate and extent of contaminant biodegradation.   These factors  were
addressed by IT during  the SITE investigation.  To  reduce the operating HRT, thereby
decreasing the size of the system, IT operated bioslurry reactors in series (i.e., plug flow)
under semicontinuous mode and evaluated two HRT set points.

 During the demonstration, IT operated two 60-liter (L) Tekno Associates  bioslurry reactors
(Salt  Lake City, Utah) and a 10-L fermentation  unit in semicontinuous, plug-flow mode for a
7-month period. The first 60-L reactor received fresh  feed daily and supplements of salicylate
and succinate to enhance PAH biodegradation.

Slurry from the first reactor was fed to the second 10-L reactor, where Fenton's reagent
(Fe+++H2C>2) was added to accelerate chemical oxidation of 4 to  6-ring PAHs.   The third
reactor in series was used to biologically oxidize contaminants remaining following addition of
Fenton's reagent. This reactor received no additions of salicylate and succinate and was
aerated, nutrient amended, and pH adjusted only.

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2.0 Conclusions and Recommendations
 2.1    Conclusions
 The 7-month demonstration illustrated the potential effectiveness of combined biological and
 chemical oxidation for the treatment of PAH-impacted soils.  Overall, the following
 conclusions were made:
       The 80 percent CPAH destruction goal was achieved with CPAH transformation
       ranging up to 84 percent.

       A system HRT of approximately 37 days increased system performance.

       Due to the increased transformation of PAH in reactor 1 (Rl) and reactor 2 (R2) during
       optimal performance, transformation rates in reactor 3 (R3) were significantly
       decreased.  This result may indicate that R3 is not required for effective treatment.

It should be noted that greater than 80 percent CPAH removal was achieved during the last 2
weeks  of operation, following modifications to the treatment process made during the previous
2 weeks.   These results are reflective of the effectiveness of the treatment system following
achievement of steady state operation.

The modification to the treatment system included increasing the system HRT from 18.5 to 37
days. This change resulted in a HRT in Rl equal to the previous system HRT. As a result,
PAH and CPAH removal increased in Rl and R2, with a decreasing performance in R3.
Overall, operation of Rl and R2 only was  adequate for effective treatment following an
increase in HRT.
2.2     Recommendations
Continued investigation under the process set points maintained during the final month of
system operation is recommended.  As demonstrated by the increase in PAH and CPAH
transformation during this period, reduced solids loading, increased clay content, and extended
HRT set points proved beneficial to the treatment process.

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Modifications to the pilot-scale reactor design, decreasing the incidence of foaming should be
investigated. Process foaming, particularly when operating on the full-scale, will result in
poor system performance, reactor overflow, and the inability to  effectively aerate the system.

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3.0 Treatability Study Approach
 3.1    Test Objectives
 The primary objective of this SITE investigation was to document accelerated PAH removal
 rates using combined chemical and biological treatment techniques.  The project used specific
 organic (succinate and salicylate) and inorganic nutrient (ammonia and phosphate)
 supplements, in combination with Fenton's reagent to achieve increased removal in a plug-
 flow treatment system.

 The  specific objectives of the pilot-scale demonstration were:

       Determine efficacy  of achieving greater than 80 percent reduction in CPAH due to a
       combination of biological and chemical oxidation

       Estimate HRT required for  operation of each reactor
       Determine CPAH reduction in each reactor to compare combined biological/chemical
       oxidation to biological treatment alone

       Determine the need for R3

       Generate performance data  upon which full-scale design can be established

       Provide operating data from which full-scale cost estimates can be generated.

 3.2    Experimental Design and Procedures
Previous  work by IT demonstrated that sodium salicylate and sodium succinate enhanced the
 levels of naphthalenedegrading bacteria in a slurry reactor.  Previously published research has
 demonstrated that in MGP site soils naphthalene, phenanthrene, anthracene, and to  a limited
extent benzo(a)pyrene [B(a)P] were mineralized (Sanseverino et al., 1993). Biodegradability
was  confirmed with 14C-radiolabeled PAH.   Further; the naphthalene-degrading pathway of
Pseudomonas purida NAH7 and NAH7-like  organisms  mineralize phenanthrene and anthracene
through the same genetic and biochemical pathway as naphthalene.  Therefore, the  presence of
salicylate will keep  the naphthalene pathway induced, promoting degradation of these 3-ring
PAHs even when naphthalene levels diminish (Ogunseitan et al., 1991).

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The primary objective of Rl operation was to increase the biological removal of organic
 carbon.  Salicylate was used to induce the naphthalene degradation operon on NAH plasmids.
 It was assumed that NAH plasmids were naturally occurring in microbial populations
 indigenous to subject soils. Succinate, a by-product of naphthalene metabolism, served as a
 general carbon source  in Rl which removed easily degradable carbon and increased biological
 activity against more recalcitrant PAH (i.e., 4-ring compounds and higher).

 Slurry from Rl was fed to R2 where Fenton's reagent was continuously introduced, resulting
 in chemical oxidation being the primary mechanism for PAH transformation in this reactor.
 The pH in R2 was adjusted to 2.0 following the addition of Rl slurry.  Fenton's  reagent
 (hydrogen peroxide in the presence of reduced iron salts) produces free radicals, which have
 been shown effective in extensively oxidizing multiring aromatic hydrocarbons in both soil and
 water systems (Gauger et al. 1990; Elizardo 1991). The objective of Fenton's reagent addition
 was not PAH mineralization, but the  hydroxylation of PAH, because hydroxylation of high-
 molecular-weight PAHs generally is the rate-limiting step in biological oxidation.

R3 was used for biological oxidation of R2 slurry.  R3 received no additions of salicylate and
 succinate. The reactor was aerated, nutrient amended, and pH adjusted following the
introduction of R2 feed.  The system process flow diagram (PFD) is  presented in Figure 3-1.

3.3 Equipment and Materials
All treatability testing was completed at  the BAG laboratory located in Knoxville, Tennessee.
This facility holds a special exemption from the State of Tennessee that permits execution of
treatability studies.  The BAG laboratory operates  in accordance with  an approved Chemical
Hygiene Plan (CHP).   All project activities at the BAG conformed to  the standards set forth in
the CHP.

3.3.1  Soil Collection and Preparation
Soils were excavated by the on-site contractor using a rubber-tire backhoe and a Kamatso
DC200 trackhoe.  Lightly impacted soils were collected from the A-cell area during site
preparation of a land treatment cell.  Highly contaminated soils were collected from Catch All
Pond Sediments in the  area where the  SB-5  sample had been collected.  Appendix B contains
all data obtained from the wood-treating  facility prior to soil collection.

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The objectives of field screening were:

    •  Prepare adequate volume of -30 mesh soil slurry to complete the pilot-scale
       investigation
    •  Dispose all oversized material on-site
    •  Prepare one 55-gallon drum of lightly impacted slurry
    •  Prepare four 55-gallon drums of a 50:50 blend of lightly and highly impacted material
    •  Collect additional volume of unscreened soil to support on-going project activities.

All objectives were met during field activities.  Soils were screened in accordance with the
approved Test Plan (IT, 1994). Site soils were excavated and staged on visqueen. Soils were
transported to the wet-sieving area using a wheelbarrow.  Three galvanized aluminum watering
troughs with stainless-steel mesh sieves secured to the rim with lumber were used during soil
screening. Each sieve was constructed using a -30 mesh U.S.A.  Standard Testing Sieve.

During screening, the troughs were partially filled with tap water. Little Giant 2E Series
submersible pumps (aluminum housing, epoxy coating, nylon pump head and  impeller, and
polypropylene screen) were placed on concrete blocks inside each trough.  These  pumps were
used to recirculate the wash water  and, thereby, increase the slurry density of the mixture.
Evaporation of excess water could  not be achieved during screening due to the limited
equipment.

Soils were characterized as sand (30 percent) and clay (70 percent).  Particle size  distribution
data are presented in Appendix A.

During screening, five drums of sieved material were generated - one drum of clean soil and
four drums of blended material.  Blended slurry was produced through the separate screening
of lightly and highly impacted soils.  The slurry produced during soil screening was then
blended in a 1: 1 ratio. In addition  to  soil slurry, two drums of impacted soil were collected.
All seven drums were shipped to the IT Bear Creek Facility located in Knoxville,  Tennessee.

Following soil collection and screening, three drums of slurry (Drums 1, 2, and 3) were
individually mixed and separated into smaller, more manageable containers. Cross-
contamination of drum contents was avoided during mixing.  A MQ Multiquip Whiteman
cement mixer with a Honda GX240 8.0 motor operated at approximately 28 revolutions per

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minute (rpm) was used to mix all slurry. The system was decontaminated using Mi-T-M
pressure washer with a Honda GX160 5.5 motor.

3.3.2      Reactor Description and Operation
Rl and R3 were 60-L,  stainless-steel, Tekno™ (formerly Eimco) bioslurry reactors (Figure 3-
2; Salt Lake City, Utah).   R2 was a 30-L glass vessel fitted with an overhead impeller
system. The system PFD is shown in Figure 3-1. Initial operational setpoints for R1,R2, and
R3 are provided in Table 3-1.  All portions of the reactor system that contacted the slurry
mixture were stainless-steel, glass, or Viton tubing.

Agitation and aeration in Rl and R3 were accomplished using a combination of the Tekno™
reactor impeller, air lift, and diffuser systems.   System pH was maintained through manual
additions of 10 Normal (N)  sodium hydroxide to the reaction vessels as necessary.

Fresh feed was manually introduced to  Rl at an average daily flow rate of 6 L/day.  For the
first 4 months of operation, fresh feed was introduced to the system in 2-L batches three times
per day to equalize the load of carbon into the system.  At this daily flow rate and operating
volume of 57 L, the Rl HRT was maintained at approximately 10 days.

Slurry from Rl was manually fed to R2.  The working volume of this 10-L reactor was
approximately 6 L. With an influent flow rate of 6 L/day, the HRT in R2 was 1 day. In
addition to the influent slurry, Fenton's reagent was added to this reactor at a rate of 2 L/day.
Overall, 8 L/day of slurry was  removed from R2 and introduced to R3.

Slurry from R2 was manually fed to R3.  The working volume of this reactor was equal to Rl.
The resulting HRT at  8 L/day was 7.5 days.  Volume loss due to evaporation was checked
daily and adjusted as needed. Effluent from R3 was collected and stored for disposal at a
licensed treatment storage and  disposal  facility (TSDF).

After 4 months of operation, the HRT was doubled to 20 days in Rl, 2 days in R2, and 15
days in R3 to reduce the loading of organic carbon on the system.  The total system HRT was
increased from 18.5 to 37 days. In addition, influent feed (3 L) was introduced once per day.

For the first 4 months of operation, Fenton's  reagent was prepared by mixing a 1:1  ratio of 35
percent hydrogen peroxide  and 1.5 molar (M) ferrous sulfate heptahydrate solution.   For  the

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 remainder of the study, Fenton's reagent was prepared using 8.8 milliMolar (mM) ferrous
 sulfate heptahydrate.  The change in the concentration of Fenton's reagent addition was
 initiated to increase the efficiency of the chemical reaction.  A reduced concentration
 prevented the occurrence  of competing side reactions.

Initially, ferrous sulfate and hydrogen peroxide were added by  dripping each solution into R2
 at a rate of 1.0 L each per day.  This system was modified such that each solution was
 introduced simultaneously below the slurry surface.  This was thought to provide for better
 mixing, less splashing of  each reagent, and reduced foam production.

Mixing efficiency of the reactor solids was verified periodically during the course  of the
 investigation. Verification was  accomplished through analysis of total solids (TS)
 concentrations in samples extracted from sample ports located on the side of the bioslurry
reactors. If nonuniform mixing was evident, agitation speed, rake speed,  airlift system, or
 solids  content was adjusted.  Due to the combined application of a 1:1 hydrogen peroxide:iron
sulfate solution in R2, only the impeller system was used to maintain complete mixing of the
 soil slurry.

 3. 4    Sampling and Analysis
 The reactors were charged on September 23, 1994.  All reactors were operated  in batch
 through October 10,  1994. Initial analytical data was colkcted prior to reactor batch
 operation, during batch operation, at the initiation of semi-continuous operation, and routinely
 throughout the remainder  of the study.  Sampling dates are presented in Section 4.0.

No steps were taken to reduce biological activity in soil samples prior to testing  due to a
shortage of refrigerated storage area.  However, each influent batch was analyzed  prior to
introduction into the reactor system to accurately determine initial concentrations.

 Slurry samples were  collected  from sample port S2 on. Rl and R3 (Figure 3-1).  Grab samples
were collected from R2.   The sampling and analytical schedule for the slurry phase, as well as
volumes required for  each analysis is shown in Table 3-2.  In addition to the slurry phase
analyses, the headspace of Rl was sampled monthly for PAHs   The influent  slurry was
sampled each time a new batch was introduced into the reactors.  Influent slurry was analyzed
for PAH, total  organic carbon  (TOC), TS/volatile  solids (VS), and density.
                                            10

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3.4.1      Physical Analyses
A summary of all analytical methods used during this investigation is presented in Table  3-3.
Physical measurements included dissolved oxygen, total and volatile solids, pH, ammonia, and
phosphate.

3.4.2     PAH Analyses
PAHs were measured in the slurry phase of each reactor, as well as the headspace of Rl.
PAH concentrations in the slurry were determined using modified EPA Method 8310. For
PAH analysis of solids, air-dried slurry samples (10 grams) were mixed with anhydrous
sodium sulfate, placed in an extraction thimble and extracted using dichloromethane (DCM) in
a Soxhlet extractor for 16 hours.  (The dry weight of the solid phase was analyzed for weight
loss in a 105°C drying oven.) The DCM extract was  concentrated to 1 milliliters (mL) using a
Snyder column and solvent exchanged to  100 mL acetonitrile. Following extraction the
sample was analyzed using a Dionex high performance liquid chromatograph (HPLC)
equipped with an ultraviolet (UV) detector at 254 nanometer (nm).   The elution profile was
acetonitrile:water (35:65) for 1 minute followed by a  gradient to 100 percent acetonitrile over
15 minutes and held for 10 minutes.

Aqueous phase PAH analysis was conducted for the first 3 months of operation.  Aqueous
phase PAH were quantified by direct injection of the  aqueous phase into a Perk&Elmer HPLC
equipped Vydac Cjg column (Model 201TP54; Hesperia, California) with a variable
wavelength programmable  fluorescence detector (LC-240). The elution profile was
acetonitrile: water (50:50) for 2 minutes followed by a gradient to 100 percent acetonitrile over
12 minutes and held for 5 minutes.

Headspace semivolatile constituents were measured through air sampling at port Z-2 on Rl
(Figure 3-1). The air sampling train consisted of a Teflon™ probe, a 47-millimeters (mm)
Teflon™  membrane filter, and an XAD-2  sorbent sampling tube.  Headspace gases  were
pulled through the XAD-2 tube for 24 hours.  The XAD tube was  extracted with 5  mL of
acetonitrile and analyzed by HPLC.

Reactor samples were shipped to the EPA  (Cincinnati, Ohio)  for extraction and analysis by gas
chromatography/mass  spectrography (GC/MS) for confirmation of the HPLC analysis.
                                          11

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 3.4.3      Microbial Enumerations
Total heterotrophic bacteria were quantified using BAG standard operating procedure (SOP)
009. Naphthalene  degrading bacteria were quantified by DNA:DNA colony  hybridization
using the NAH gene as a probe (Sayler et al., 1985; Sanseverino et al.,  1993). The NAH
encodes the naphthalene dioxygenase enzyme which is the first step in naphthalene
metabolism. Enumeration of naphthalene-degrading bacteria occurred on weekly samples for
the first three months of operation at which time it was discontinued.

3.4.4      14CMineralization Assays
  C-PAH  mineralization assays were performed to estimate in situ microbial degradative
capacity for specific  compounds. Two mL of slurry were placed in a 40-mL vial (Pierce,
Rockford,  Ii.). Labeled l-14C-naphthalene, 9-14C-phenanthrene, UL-14C-anthracene, or 7, 10-
14C-benzo(a)pyrene (Sigma, St. Louis,  Mo; specific activity 8.0, 10.4, 10.4, and 60.0
mCi/mmol,  respectively) were individually added to triplicate vials.  Slurries were incubated at
26°C with  shaking (100 rpm). Naphthalene and phenanthrene mineralization  (  C02
production) was analyzed at regular intervals over a 7-day period.  Anthracene mineralization
was analyzed at intervals up to  10  days. Benzo(a)pyrene mineralization was analyzed at
intervals  up to 2 weeks. Biological control samples were inhibited by acidification with 0.5
mL of 2 N  sulfuric acid (HzSO^) and metabolism assays were also terminated by H2S04
addition.  14C02 was  trapped in  0.5 mL of 0.5 N NaOH. The NaOH was added to 1 mL of
water and 10 mL of Beckman ReadySafe m scintillation fluid and counted in a Beckman liquid
scintillation counter (Model LS380 1).

 3.4.5      Toxicity Screening
The aqueous phases  of the influent  feed, Rl, and R3 were tested for toxicity using
Terrahymena as a test organism.   Dilutions of the aqueous phase (1:200, 1: 100, 1:40) were
added to Tetrahymena cells (approximately 1,000 cells/ml)  and incubated for 16 hours. Acute
toxicity was assessed by observing  each tube for cell lysis and/or lack of ciliary movement as
determined with the aid of a dissecting microscope.  A lethal dose (LDso) was determined for
each sample.

3.4.6      Chemicals
Chemicals,  analytical grade, and sources are listed in Table 3-4.
                                           12

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3.5      Data Management
All data was generated and stored as specified in the approved Test Plan

3.6     Deviations from the Test Plan and QAPP
Per the project contract, work progress and quality were monitored through audits of the
laboratory and project files.  Audits indicated minor nonconformances and technical changes
from the Test Plan, however, nothing was identified which adversely impacted the project
quality of work.  All project technical  changes and nonconformances are listed in Tables 3-5
and 3-6.
                                           13

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4.0 Results and Discussion
4.1     Data Analysis and Interpretation
The results of the 7-month demonstration are presented in the following text.

4.1.1      Reactor Operation
Daily reactor operation was described in Section 3.3.2.   Operational difficulties  were
encountered, during the demonstration.  First, tar balls were formed following reactor charging
and aeration.  These tar balls were physically removed from the system and their mass
determined. Following removal, PAH concentrations and mass within all reactors were
recalculated to assure that physical removal was not accounted for as biological/chemical
removal.

Second, foaming of the reactor contents was a routine occurrence.  Rl and R3 were fitted
with mechanical foam breakers but these were inadequate to contain the foam.  Antifoam  289
(Sigma, St. Louis, Missouri) was used as required to  contain the foam inside  the reactors.

The third problem was clogging of the airlifts.  This problem usually  appeared due to
disruption of the air flow to the reactors and unusually high settling due to  decreased clay
content of the soil. Periodic purging and/or dismantling  of the air lifts were  necessary to
restore air flow.

Table 4-1 summarizes significant upsets and changes  which effected reactor operations.

4.1.2     Physical A nalyses
Results of physical analyses are presented below

4.1.2.1     pH
The pH of Rl and R3 were maintained at 7.0 (Figure 4-1) through the addition of NaOH  on
an as needed basis.  Rl required minor periodic adjustments in pH through the addition of 1
N NaOH.   The material transferred from R2 into R3 required  pH adjustment with every
transfer using 10 N NaOH.  The Fenton's reagent dictated the pH in R2. When 1.5 M
                                           14

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ferrous sulfate was used, the pH ranged from 2.0 to 2.5. When 8.8 mM ferrous sulfate was
used, the pH reached a new steady state at 3.5 to 4.0.

4.1.2.2    Dissolved Oxygen
For the first 3 months of operation, the dissolved oxygen concentration fluctuated between 0
and  10 milligrams per liter (mg/L) (Figure 4-2).  Due to foaming within the reactors, it was
not always possible to obtain a "pure" liquid sample for dissolved oxygen analysis.   The last 3
months of operation showed a steady dissolved oxygen in Rl ranging from 2.0 to 7.0 mg/L.
The  dissolved oxygen in R3 ranged from  4 .0 to 9.0  mg/L over the same period.

4.1.2.3    Reactor  Solids
The reactors were initially charged at 40  percent solids using the  highly contaminated site soil.
This loading was necessary for the airlifts to operate properly and keep the solids in suspension
due to high sand content.   Due to increased foaming problems, the impacted TS content was
reduced to 20 percent and Celatom    diatomaceous earth (DE)  was used to increase the TS
concentration to 35 percent.  This switch occurred on December  13, 1995. Use of DE was
discontinued on December 29,  1995  and  locally obtained clean clay was used to reduce the
influent  organic content.   The solids loading for the final 3 months of operation was 30
percent contaminated soil plus  10 percent clay for a 40 percent total solids loading.

Figure  4-3 shows the TS for  each reactor.  For the final 3 months of operation, the  TS in  Rl
was  steady at 30 to 35 percent. R2 showed a decrease in TS relative to Rl. This decrease was
due to dilution of the reactor  contents with Fenton's reagent.  R2 and R3 showed  a  steady
decline in TS over time.  On January 4,  1995, the TS was 29  percent. On April  19, 1995, the
TS in R3 reached a low of 9  percent.

Volatile solids for  each reactor are shown in Figure 4-4.  Rl VS remained steady for the last 2
months of operation ranging from 8 to 9 percent. R2 displayed wide fluctuations  in VS
ranging from 5  to  14 percent. R3 showed an increase in VS with values ranging from 10 to  13
percent over the last 2 months  of operation.  The increase in VS  may represent increased
biological growth possibly due to the metabolism of recalcitrant hydrocarbons oxidized in R2.

The VS/TS ratio (Figure 4-5) remained steady for Rl ranging from 0.2 to 0.3 while R3
showed a  steadily increasing VS/TS  ratio during the final 2 months of operation.  R3 values
                                           15

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 ranged from  0.8 during early March to  1.3  on April 19,  1995. R2   VS/TS values fluctuated,
 but centered around  0.4 to 0.5.

 Solids  distribution for Rl and R3  is shown  in Figures 4-6  and 4-7,  respectively.  A uniform
 solids  suspension was dependent on the airlift operating properly, maintaining  an adequate
 solids  loading  within the reactor,  and  maintaining  the proper  volume.   Stratification  was  a
 minor  problem in Rl (Figure 4-6) but was a more persistent problem in  R3 (Figure  4-7).   The
 DE promoted stratification by binding  to the clay  present in the contaminated soil.  This  did
 not pose a significant problem in  Rl due to  the high  organics  present, however, it may have
 been a problem in  R3 as seen in the solids distribution during the months of December, 1994
 and January,  1995.

 4.1.2.4      Nutrients
 Ammonia was added initially once per week for the first 2 months followed by 3 times per
 week  for the remainder of the study.   Ammonia  addition to R3 was  discontinued  after 3
 months due to carry over from  Rl and R2.  Ammonia values  ranged from 0 to 240  mg/L over
 the course  of the study (Figure 4-8).

The large increase in ammonia concentrations resulted from increased addition rather than
 reduced utilization.  During the mid project review meeting held in December it was  decided
 that a  possible nitrogen  deficiency may have resulted in system foaming. As a result, the
 nitrogen addition rate to the system  was increased.

 Phosphate was initially  added for the first  month of operation  at which time it was
 discontinued.    The phosphate concentration  in the contaminated soil was  significant  and
 ranged from  100  to 200  mg/L in  each reactor  during  the course of the investigation (Figure 4-
 9).

 4.1.2.5    Microbial  Enumerations
 The total heterotrophic  bacterial  populations  in Rl ranged from 1.4  x 10'  colony forming units
 per mL of slurry (cfu/mL)  to 4.0  x  1$  cfu/mL  slurry over the course of the  study (Figure 4-
 10).  Bacterial  populations in R3 were similar to  Rl  except  during the last 2 months of the
 study.  Total populations  reached  2.2  x 109 cfu/mL slurry on  March  2,  1995  and  ranged  from
 3.0 to  5.0 x  10s cfu/mL slurry for the remainder of the study.   This increase  coincided with an
 increase in the HRT  set  points.

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 The naphthalene-degrading bacterial populations  were 3.6  x  105 cfu/mL  slurry at the time the
 reactors were  charged. This population dropped  below the method detection limit (2.0  x 105
 cfu/mL)  after onset of continuous reactor operation.   Naphthalene-degrading  bacteria  were
 enumerated  by colony  hybridization in the April 13,  1995 samples.    The influent  feed
 material, Rl,  and R3 contained 1.5 xlO4, 1.1 x 106, and 1.1 x 108NAH positive cfu/mL
 slurry.  The large increase in NAH positive cells in R3 was surprising since there was  no
 detectable naphthalene  in R3.

 4.1.2.6      TotalRecovemblePetroleum Hydrocarbons
 Total recoverable petroleum hydrocarbons (TRPH) were  measured in March  16,  1995
 samples.  The data is summarized in Table 4-2.  R2 had  no apparent effect on  TRPH while R3
 showed a 86  percent reduction.  Even though R2 had the same TRPH concentration  as  Rl, this
 does not imply that the TRPH is in the same form as in  Rl.  ERA Method 418.1 does  not
 discriminate differences in  hydrocarbon chain length or possible  side  chain modifications (such
 as  hydroxylations).  Therefore, the  Fenton's reagent  could have broken  down longer chain
 hydrocarbons into  shorter chains which were more susceptible to bacterial  degradation in R3.

 4.1.2.7     Total  Organic  Carbon Analysis
 Solid and  aqueous phase TOC concentrations are  shown in Figures  4-11 and 4-12,
 respectively.    The average  influent  solid phase total  carbon  (TC) concentration  was 19,000
 mg/kg. On  March 16, 1995, influent  solid phase TC was 17,500 mg/kg. After treatment,
 there was  a  55 percent reduction in TC.   R2 showed a 48 percent reduction relative to Rl.
 Although this  reduction  was not consistent with the TRPH removals,  there may have been
 shorter chained organic compounds not measured in the TRPH analyses  which were
 mineralized  in  R2.

 For the 4-week  period from March 16 through  April 19,  1995, the average solid  phase  TC for
 Rl, R2, and R3 was 13,700; 7,900; and 6,400 mg/kg, respectively. In comparison, for the
 first 6 months  of the study, the average solid phase  TC for Rl, R2, and R3 was  14,500;
 11,900; and 10,000  mg/kg, respectively.

The average aqueous phase TOC for the  4-week period from March 16 through April 19, 1995
in Rl, R2, and R3 was 570; 1,680; and  740 mg/L, respectively.  In comparison, for the first 6
 months of the  study, the average aqueous  phase TOC in  Rl, R2,  and R3,  was 690;  1,680; and

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340  mg/L,  respectively.  By observation of Figure 4-12, the  production of aqueous phase  TOC
 was erratic in R2 while  Rl was consistent over the 7-month period.

 4.1.2.8    PAH and Mass Balance
 Physical removal  of PAHs  during treatment were  corrected during the  determination  of mass
 removal.   All tar ball removal  was corrected  through  the recalculation of initial PAH
 concentrations and mass  in  all  reactors following physical removal.   Significant wall  losses
 were not  recognized following the dismantling  of the reactors due to operation at  all the
 reactor's  full  working volume.   Dilution with the addition  of Fenton's  reagent was corrected
 by using  real-time TS concentration data from each reactor to determine PAH mass.

 Additionally the difference in sampling location  for  Rl and R3 as compared to R2 did not
 create a sampling bias in PAH  results.  All reactors were tested for adequate mixing  and  PAH
 mass determinations were corrected for the TS concentration of the sample.   Reduced
 performance in R3 during the final stages of the testing  program were due to the increased
 performance of  Rl and R2 and reduced  influent  carbon concentrations in  the reactor.

 It should also be noted that  PAH  percent removal  perturbations were due to operational
 conditions rather than a decrease in PAH recovery efficiencies.  See Section 4.2 for  a detailed
 discussion.

 Slurry samples from  each reactor were dried and analyzed by modified  EPA method  8310.
 This method accounted for  any  recoverable PAH in the soil and aqueous phases.   The air
 phase was monitored once per month  and no  substantial volatilization of PAH was  observed in
 Rl.

 The bioslurry  reactor system  demonstrated 95  and 84  percent removal of  PAH and CPAH,
 respectively, as  of April  19,  1995. Figures 4-13 through 4-16 illustrate  PAH  and CPAH
 reductions in Rl, R2, R3,  and overall. Overall, the biologically  active reactors (Rl and R3)
 illustrated a decreasing  effectiveness in PAH  transformation as a function  of compound
 molecular  weight.  This is  indicated in Figures 4-17 and 4-18 which present the concentrations
 of fluorene and  B(a)P throughout the  system.

 Prior to operational changes  initiated  in  March  (following 5 months of treatment),  Rl
 demonstrated 62 percent  transformation of PAH, with approximately  28 percent transformation

                                              18

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of CPAH.  R2 demonstrated comparable destruction  of PAH and CPAH (approximately 30
percent), as expected  during  chemical  oxidation. R3  CPAH and PAH  transformations
averaged approximately  14 and 33 percent, respectively. The total  system PAH and CPAH
transformations averaged 85  and  65  percent,  respectively.  All PAH  and CPAH  removal
efficiency data are presented  in Tables 4-3  and 4-4, respectively.   No  significant volatilization
of PAH was evident in  Rl.

Following operational  changes initiated in March, overall PAH and CPAH transformation
rates increased up to 95 and  84 percent.  Rl  demonstrated  87 percent transformation of PAH,
with 65  percent transformation of CPAH. R2 demonstrated comparable destruction  of PAH
and CPAH (greater than 45 percent),  as  expected during chemical  oxidation. R3  CPAH  and
PAH  transformations were decreased  averaging -31.6  and  -26 (0)  percent,  respectively.  The
total system PAH  and  CPAH transformations  increased to  91 and 75 percent, respectively.
All PAH and CPAH mass removal efficiency  data are presented in  Tables 4-5 and 4-6,
respectively.

During  optimal operation,  the  influent  PAH concentration  was  decreased from 6,210  mg/kg  to
325 mg/kg. Influent CPAH concentrations were decreased from 422  mg/kg to 65 mg/kg.
CPAH and PAH concentrations throughout the system are presented in  Figures 4-19 and 4-20,
respectively. All PAH data is presented in Appendix C.

Mass  balance  data is  presented in Appendix  C.   All  mass  removals  were calculated per the
specifications  of the approved Test Plan Table 4-3.  This table  has been included in Appendix
C for reference.  PAH and CPAH mass balance data has been summarized  in Tables 4-5 and
4-6.  Over  the  7-month operational  period,  Rl demonstrated the highest transformation  of
PAH, averaging 10.6 grams/day (g/day). Following March 2, 1995, the PAH mass removal
in Rl was  increased to 7.7 g/day.

R2 and R3 demonstrated  3.2  and 3.2  g/day  PAH removal,  respectively, during the entire
operational  period (Table 4-5). R2 and R3  PAH removal efficiencies also decreased following
March  2, 1995. R2 and R3  demonstrated 1.17  and  0.48 g/day  PAH  removal,  respectively,
from March 2  to Apnl  19, 1995.

CPAH mass removal is summarized in Table 4-6.   The same trends  in PAH removal were also
evident  when analyzing  the CPAH removal data.  CPAH mass removal in Rl, R2, and R3

                                            19

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during the total operational period averaged  0.93,0.44,  and  0.24  g/day, respectively.   Rl
performance following  March 2, 1995  indicated an average CPAH removal of 0.63 R2 and  R3
CPAH removal was decreased following  March 2, 1995  to an average  of 0.22 and 0.08 g/day,
respectively.

Since February 2, 1995, R3 showed  little  to  no transformation of PAH.  In fact, there was an
increase  in PAH  observed in the slurry.   This  observation was preceded by the change in
addition  of  Fenton's reagent to  R2.  It is  also  known (at  least for March 16, 1995 samples) that
62 percent of the total  TRPH was removed by  R3.   It is  hypothesized that if R.2  was breaking
down the larger chain  aliphatic hydrocarbons to smaller chain  aliphatic hydrocarbons and these
hydrocarbons  were  subsequently metabolized  in R3, then  PAH could be released  from these
aliphatic  hydrocarbons.  The tar phase, not the soil, is  considered to be the dominant phase in
manufactured  gas plant sites and  creosote-contaminated  sites (Lane  and  Loehr,  1993).
Therefore, all  PAH will be found  sorbed to the tar.  If the tar phase breaks down, the sorbed
PAH  will be released. The build-up in R3 may reflect PAH being released from the tar phase
and accumulating in the reactor.  Whether PAHs were  being metabolized in R3 is not clear
although  14C-mineralization data discussed in Section 4.1.2.9 suggests that at least the   2- and
3-ring  PAHs  were  metabolized.

4.1.2.9     l4C-Mineralization  Assays
First-order mineralization  rates were  determined by calculating the  available (soluble)  PAH for
degradation  and calculating specific activities  for each   14C-PAH  in  each soil.  To estimate  the
distribution  of each PAH  in the aqueous phase, the tar-water partition coefficient  (Ktw) was
estimated (Lane and Loehr 1992).  The tar phase,  rather than the particulate phase, is the
dominant force in  determining partition in  these soils due  to the  high  organic carbon content.
Lane and Loehr (1992),  using MGP soils in their experimental design, determined that a
relationship  existed  between the octanol-water partition  coefficient (KoW) and  K^.

                   log Ktw =  1.13  log Kow  + 0.33               (Equation  1)

The log Rvalues for naphthalene,  phenanthrene and anthracene were 3.37, 4.46, and 4.45,
respectively  (Sims and  Overcash  1983). Specific activities of  PAH  were determined by
dividing the  /iCi  of specific radiolabeled PAH added to the slurry by the calculated number of
jtmoles of specific  PAH present in the aqueous phase.   Mineralization  rates  were determined
                                            20

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 by plotting the number of jimoles of 14CC>2 produced vs time.   The  slope of the initial linear
 portion (r2 * 0.95) of each curve was used as an estimate of the mineralization rate.

 4.1.2.10  Toxicity Screening
 To estimate the reduction in toxicity of the aqueous phase, an  assay  using the protozoan
 Tetrahymena was performed.   Table 4-7  summarizes the results with an estimate  of the lethal
 dose-50 (LDso)  for the aqueous phases of the  influent feed,  Rl, and R3.  Rl  and R3 had
 at  1: 100 dilution of the  aqueous phase in comparison to an LDso at the  1:40 dilution for the
 aqueous phase of the  influent  feed.  No definitive  conclusions  should be drawn from this
 assay. While  there was a substantial reduction in PAH  and TRPH, pentachlorophenol   (PCP)
 and arsenic were  also present  in this soil.  Although PCP was not  measured  directly,  past
 experience has shown that there  is little to no reduction  in this type of treatment  system.
 After passage through R2, Fenton's reagent should  be in its  most oxidized form  (As+5).

 4.2    Quality Assurance/Quality Control
 GC/MS analyses  confirmed the  correct identification of all PAH  peaks  measured using the
 HPLC.  In addition,  an  approximate 20  percent variance comparing the  GC/MS and HPLC
 analyses was  noted.  All other analytical measurements  were made within the specifications
 defined in the approved  quality  assurance project plan (QAPP) unless specified in Table  3-6.

With  regard to accuracy  criteria  for  matrix spikes using modified Method 8310, the stipulated
 acceptance criteria provided in Table 6-1 of the QAPP was a minimum  recovery  of 80  percent
 for each of the 16 compounds.  The corrective  action for those matrix spikes  which did  not
 meet criteria  was reanalysis of the sample extract.   Upon analysis and  reanalysis,  where
 necessary, 95 percent of the matrix spike recoveries met this  criteria, and nonconformances
 were generated for any outlying  data points.

4.3  Costs/Schedule for Performing the Treatability Study
 The overall budget for project execution was $209,751,   All project  activities were conducted
 between September 1994 and  October 1995.

4.4    Key   Contacts
 The EPA  Project  Manager, Brunilida Davila, can  be  contacted at (513-569-7849). IT contacts
 including the project manager  (Kandi Brown)  and principal investigator (John  Sanseverino)
 who  can be reached at 909-799-6869 and  615-690-3211  respectively.

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5.0   References
Brown,  K.L.  and J.  Sanseverino,  1993,  "Factors Affecting PAH  Biodegradation in
Liquid/Solids  Contact Reactors,  " Proceedings of the  86th  Annual Meeting and Exhibition of
the Air and Waste  Management Association,  Denver,  Colorado.

Davila, B. F. Kawahara, and J.  Ireland, 1994, "Combining Biodegradation and Fenton's
Reagent to  Treat  Creosote Contaminated Soil," In Press.

Elizardo, K.,  1991,  "Fighting Pollution  with  Hydrogen Peroxide,"  Pollution  Engineering,  pp.
106-109.

Gauger, W.K., V.J.  Srivastava, T.D. Hayes, and D.G. Lrnz, 1990, "Enhanced
Biodegradation  of Polyaromatic  Hydrocarbons in Manufactured Gas Plant  Wastes,"
Environmental   Biotechnology.

IT Corporation,  1994, SITE  Emerging Technologies Program  E06  - Bioslurry Treatment Test
Plan,  July.

Lane  W.F.  and Loehr  R.C., 1992, "Estimating the Equilibrium Aqueous  Concentrations  of
Polynuclear  Aromatic Hydrocarbons  in  Complex Mixtures",  Environmental  Science and
Technology,  26:983-990.

Ogunseitan, O.A., I.L.  Delgado, Y.-L. Tsai, and B.H. Olson, 1991, "Effect of 2-
Hydroxybenzoate  on  the Maintenance of Naphthalene-Degrading Pseudomonads in Seeded and
Unseeded  Soil", Applied and Environmental Microbiology,  57:2873-2879.

Sanseverino J, B.M. Applegate, J.M.H. King, and G.S. Sayler, 1993, "Plasmid-mediated
Mineralization  of Naphthalene,  Phenanthrene, and  Anthracene," Applied  and Environmental
Microbiology,  59:  1931-1937.

Sayler,  G.S., M.S. Shields,  E.T.  Tedford,  A. Breen,  S.W. Hooper, K.M.  Srrotkrn, and J.W.
Davis, 1985,  "Application of DNA-DNA Colony Hybridization to  the  Detection of Catabohc
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Genotypes in Environmental  Samples, "  Applied  and  Environmental  Microbiology, 49:  1295-1303.

Sims, R.C. andM.R. Overcash, 1983, "Fate of Polynuclear Aromatic Hydrocarbons (PNAs)
in Soil-plant  Systems," Residue Review, 88:  1-68.

U.S.  Environmental Protection Agency,  1990, "Slurry  Biodegradation,  " EPA/540/290/016
                                          23

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                                           Table 1-1
                    Physical/Chemical Properties of CPAH Constituents


                                    IT Project No. 408250

Carcinogenic Polynuclear
Aromatic Hydrocarbons
benz(a)anthracene
benzo(a)pyrene
benzo(b)fluoranthene
benzo(k)fluoranthene
chrysene
dibenz(a,h)anthracene
benzo(g,h,i)perylene
indeno(l ,2,3-c,d)pyrene

Kow
Cog)
5.61
6.04
6.57
6.84
5.61
5.97
7.23
7.66
Aqueous
Solubility
(MR/0
14
3.8
1.2
0.55
2
.50
0.26
62
V.P at
20°C
(torr)
S.OxlO'9
S.OxlO'7
S.OxlO'7
S.OxlO'7
6.3xlO'7
l.OxlO40
l.OxlO40
l.OxlO40
Sims, R. C. and M, R. Overcash "Fate of Polynuclear
Aromatic Compounds (PNAs) in Soil - Plant Systems," Residue Reviews, 1983.

jig/1 - micrograms per liter
V.P. - vapor pressure
                                              24

-------
                       Table 1-2
        General  Influent  Feed Characteristics
               for Bioslurry  Treatment

                IT Project No. 408250
   Parameter
         Target
    Oiganics
     Solids
     Water
Solids Particle Size
   Temperature
      PH
0.025 -25 percent by weight
 10-40 percent by weight
 60 - 90 percent by weight
    Less than 1/4  inch
        15-35°C
        4.5 - 8.8
        EPA, 1990, "Slurry Biodegradation,' EPA/540/290/016
                           25

-------
         Table 3-1
Initial Operational Set points

   IT Project No. 408250
Parameter
Feed Flow
Hydraulic Retention Time
Temperature
Dissolved Oxygen
oH
Agitation
Working Volume
Ammoniacal Nitrogen
0-phosphate
Sodium Salicvlate
Sodium Succinate
Fenton's Reagent
Addition
Rl
6L/day
10 days
25°C ± 5°C
3 mg/L
7.0 ±05
500 rom
57 L
50 mg/L
10 mg/L
100 me/L
10 mg/L

R2
6L/day
1 day
24°C + 5°C
...
<5.0
250 rpm
6L
---
___
«.«.«.
--_
2L/day
R3
8L/day
7.5 days
25°C ± 5°C
3 mg/L
7,04:0.5
500 rpm
57 L
=»*»
10 mc/L

—«.
-..'
            26

-------
                          Table 3-2
Sampling and Analytical Schedule for Rl, R2, and R3 Slurry Phase

                    IT Project No. 408250



Volume
(mL)
Frequency

Volume
(mL)
Frequency
Analysis
PAH

100
1/wk
TOC N&P

100
1/wk

100
1/wk
DO
First 1
1
2/wk
PH
TS/VS
Density
Microbial
Enumerations
Particle
Size
"our Months of Operation
-
daily
20
2/wk
-
daily
10
1/wk
500
1 /month
(Rl only)
Last Two Months of Operation
too
Biweekly
100
biweekly
100
1/wk
1
2/wk
-
daily
20
2/wk
-
daily
10
biweekly
500
1 /month
rat nnlvt
                             27

-------
           Table  3-3
Summary of Analytical  Methods

     IT Project No.  408250
Parameter
PAH
PAH
TOC
TC
TS
VS
NH3
o-PO4
Total
Heterotrophs
Naphthalene
Degraders
PH
DO
Particle Size
PAH
Sample Type
soil/water
soil
waler
soil
slurry
slurry
flurry
slurry
slurry
flurry
flurry
flurry
flurry
headspace
Method Number
modified EPA Standard
Method 8310
EPA Method 8270
BAC008
BAC031
Standard Methods
2540G or 2540B
Standard Methods
2540G or 2540B
BAC022
BAC015
BAC009
NA
BAC014
BAC021
ASTM Method D422
modified NIOSH 5506
Method Title
Polynuclear Aromatic Hydrocarbons
Polynuclear Aromatic Hydrocarbons
Carbon Analysis Using the Dohrmann Total
Carbon Analyzer
Total Carbon Analysis
Total, Fixed, and Volatile Solids in Solid and
Semisolid Samples or Total Solidr Dried at
103-105°C, respectively
Total, Fixed, and Volatile Solidr in Solid and
Semisolid Samples or Total Solids Dried at
103-105°C, respectively
Electronic Ammonia Analysis
Phosphate Analysis
Microbial Enumeration Analysis
Application of DNA-DNA Colony
Hybridization to Ihe Detection of Catabolic
Genotypes in Environmental Samples
pH Analysis
Oxygen Analysis
Particle Size Analysis
Polynuclear Aromatic Hydrocarbons
Method Type
HPLC
GC/MS
TOC Analyzer
Persulfate
Oxidation
Drying oven
Drying oven
Ion Probe
Colorimetric
Spread Plate
Colony
Hybridization
Membrane probe
Galvanic cell
Sieve and
Hydrometer
HPLC
Reference
SW-846
SW-846
BAC Proprietary
BAC Proprietary
Standard Method
Standard Method
BAC Proprietary
BAC Proprietary
BAC Proprietary
Saylcr, et al.,
1985
BAC Proprietary
BAC Proprietary
ASTM
Standard
Methods
              28

-------
      Table 3-4
Summary of Chemicals

IT  Project No.  408250
Chemical
Sodium salicylic
Sodium succinic
Grade
Reagent
Source
J.T. Baker, Philhpsburg, N.J.
Reagent Mallinckrodt. Paris, KE.
Ammonium chloride Reagent
Potassium phosphate dibasic 1 Reagent
Hydrogen peroxide (35%)
Sodium Hvdroxide
Ferrous sulfate • 7H20
Sodium sulfate anhydrous
Methylene chloride
PAH standards
| Acetonitrile
Reagent
Reaeent
Reagent
Reagent
Nanograde

Nanograde
Mallinckrodt, Paris, KE.
J.T. Baker, Phillipsburg, NJ.
PB&S Chem. Co., Hendersonville, KE.
J.T. Baker. Philliosbure. NJ.
Sigma Chemical Co., St. Louis, MO.
Mallinckrodt, Paris, Kentucky
Burdick & Jackson, Muskegon, WI.
Supelco, Inc., Belief onte. PA.
Burdick & Jackson, Muskegon, WI.
         29

-------
                                  Table  3-5
                        List of Technical  Changes
                         IT Project No.  408250
CHANGE
 NUMBER
         TECHNICAL  CHANGE
      EPA
 APPROVED3
      1
     4
     5
     6
     7
     8
     9

     10

     11

     12

     13
     14
     15

     16

     17
BAG completion of initial  EIMCO  soil
evaluation.
Use of 1.3 M ferrous sulfate and 30  percent
peroxide  solution to  prepare Fenton's
reagent.
Only  reagent grade chemicals used are
nutrients.
Reactor 2 equipment change.
No influent air filtration.
All reactors sampled from middle port.
Calibration of thermocouple by  manufacturer.
Change in components of air sampling tram.
Placement  of anhydrous  sodium  sulfate
during sample extraction.
Vessel volume  change during initial  batch
study.
Correction  of QAPP  concerning surrogate
additions.
Addition of GC/MS  confirmation for PAH
and PCP.
Fenton's reagent addition  at 2.0  L/day.
PAH  extract  exchange and dilution.
Preparation of  surrogates  through  1-gram  vial
additions and analysis.
QA/QC analyses for PAH  at 10 percent of ail
samples collected.
Surrogate  switched to  1-Fluoronapthaiene.
    10/7/94

    10/7/94

    10/7/94

    10/7/94
    10/7/94
    10/7/94
    10/7/94
    10/7/94
    10/7/94

    10/7/94

    10/7/54

    10/7/94

    10/7/94
    10/7/94
    10/7/94

    10/7/94

Verbal on 10/7/94
                                           30

-------
          LIST OF TECHNICAL CHANGES (CONTINUED)
                          IT Project No.  408250
CHANGE
NUMBER
18
19
20
21
TECHNICAL CHANGE
Increase reactor and influent feed TS
concentration to 40 percent.
Change in salicylate and succinate
concentration additions.
Discontinued use of the clarifier.
Increased daily TS and density measurements
EPA
APPROVEDa
Verbal on
Verbal on
Verbal on
Verbal on
10/7/94
10/7/94
10/7/94
10/7/94
      22

      23
      24

      25
      26
      27

      28

      29

      30
and reduced DO measurements  during period
of varying  reactor solids distribution.
Rerun samples when matrix is out  of ±_ 20
percent
Recalculate MDLs
Changes in the  analytical schedule  for
TS/VS, DO, aqueous PAH, nitrogen, gene
probe, and pH
Increased impeller speed to control DO
Adjust volume  of Rl
Decrease influent  organic loading by one-half
through the mixing  of  clean clay
Reduction in Rl and R3 HRT from 10 to 20
days
Reduce feed introduction to  each reactor from
twice/day to once/day
Discontinued addition of salicylate  and
succinate
Verbal on 10/7/94

Verbal on 10/7/94
     12/20/94

    12/20/94
    12/20/94
    12/20/94

    12/24/95

    12/24/95

     4/6/95
a    Date  indicates date  of EPA correspondence  documenting technical variance approval.
                                      31

-------
                                   Table 3-6
                      List of  TechnicalNonconformances
                             IT Project No. 408250
NONCONFORMANCE
         NO.
     NONCONFORMANCE
  DATE
REPORTED
           1

           2
           3
           4
           5

           6
           7
Influent  feed added as 2,  3L transfers
instead of 3,  2L transfers.
Bottle study  recoveries
Influent  T0 MS and MSD
SRM  acceptance criteria
PAH not identified during  aqueous
analyses
Method  calibration  for  chrysene 10/14/94
Missing  check standards
   10/19/94

   11/8/94
   11/8/94
   12/12/94
   12/12/94

   12/12/94
   12/12/94
                                           32

-------
                      Table 4-1
Summary  of Reactor Upsets and Operational Modifications

                 IT Project No. 408250
„ - * Date ~%i
29 November, 1994
5 December, 1994
13 December, 1994
29 December, 1994
HJanuary,1995
24 January, 1995
27 February, 1995
22 March, 1995
TT . IK r VI* :' • -WWXvM-K-fftJK-W-S
UpsetsMcohcations
Rl airlifts clogged
Rl dismantled to remove clogs from airlifts; tar balls
responsible for clog
In order to reduce organic loading in Rl, solids were
reduced to 20% and 15% diatomaceous earth was
added
Discontinued use of diatomaceous earth as a solid
supplement; caused air lifts to clog
Supplemented solids with a local, clean clay
Switched to 8.8 mM FeS04.7H20
Increased HRT to 20 days for Rl , 2 days for R2, and
20 days for R3
Discontinued salicylate and succinate addition to Rl
                         33

-------
                                     Table 4-2
Summary of Total Recoverable Petroleum Hydrocarbons (TRPH), PAH, and CPAH for
                                March 16 Samples

                              IT Project No. 408250
Sample-"" '\ ,
Influent Feed
Reactor 1
Reactor 2
Reactor 3
;?>5:,,TKPH
27,000
14,000 (48%)1
14,000 (48%)
3,700(86 %)
PAH
5860
710 (88%)
430 (93%)
560 (90%)
^ICPABu
430
130 (70%)
70 (84%)
110 (74%)
  Number in parenthesis represents the percent reduction relative to the  influent feed TRPH
                                  concentration.
                                       34

-------
              Table 4-3
PAH Removal Efficiencies (Percent)

       IT Project No. 408250
Date
10/10
10/19
10/26
11/2
i 11/9
11/16
11/22
Reactor 1
85
65
29
55.5
59.2
61.7
55.3
Reactor 2
' -180
, 58.2
45.2
37.7
26.7
41
42.9
12/2 30.4 62
12/8 56.9
12/15 68.6
12/30 , 71.6
1/5
1/26
2/2
2/16
3/2
3/9
3/16
3/30
4/13
4/19
Total Operational
Period Average
Average Prior to
3/2
Average Following
3/2
63.6
69.3
71.6
79.8
85.9
87.9
86
85.8
88.6
88.3
69. 17 ±17.9
6 1.5 ±15.45
87.08 ± 1.32
44.1
~72
51.1
64
44.8
38
22.3
33
39.1
37
65.9
37
55.4
32.7 ±50.3
28 ± 58.95
44.6 ± 13.04
1
Reactor 3 I Overall
36.1 72.6
67
59.3
42
44.8
59.6
69.5
28.9
5.7
31.7
6.5
44.8
12.1
1.2
-19
-82
-31
10.3
-66
12.9
-1.1
15.87 ± 40.57
32.68 ±26.56
-26.2 ± 40.53
95.2
84.2
83.9
83.5
90.9
92.2
81
77.2
83.3
87
92.8
85.1
82.6
81.3
82.8
90.4
92
91.9
93.8
94.7
86.59 ±6. 17
84.85 ± 6.05
90.93± 4.26
      ± Indicates standard deviation.
                 35

-------
            Table 4-4
CPAH Removal Efficiencies (Percent)
       IT Project No. 408250
Date
10/10
10/19
10/26
11/2
11/9
11/16
11/22
12/2
12/8
12/15
12/30
1/5
1/26
2/2
2/16
3/2
1 3/9
3/16
3/30
4/13
1 4/19
Total Operational
Period Average
Average Prior to
. 3/2
Average Following
I1 3/2
Reactor 1
60
20.2
-16
2.5
-1.1
9.3
30.4
10.4
30.9
32.7
43.5
38
30.7
37.6
44.4
66.4
71
59.9
60.9
69
64.8
36.83 ±25. 18
27.99 ±22.11
65.33 ± 4.39
Reactor 2
-58.2
56.9
48.5
40.5
23.9
50
33.4
51
48.3
15
43.3
59
35.3
40
23.3
34
46.3
44
69
44
55.9
38.26 ±25.52
34.01 ±27.51
48.87 ±12.08
Reactor 3
40.9
49
33.9
26
44
44.5
51.2
26.3
-7.1
1 36.1
!
12.9
-26
14.4
-8.4
-5
-105
-63
2.5
-93
-1.2
-3.1
3.3 ±44. 19
14.23 ± 39.51
-31.56 ±43.75
Overall
62.2
84.3
60.5
56.9
56.9
74.7
77.4
67.7
61.8
63.4
72.1
67.7
61.6
59.4
55.2
54.2
74.7
7S \
76.6
82.6
84
68.19 ± 9.92
65.45 ±8.42
75.02 ±10.79
-1- Indicates standard deviation.
               36

-------
            Table 4-5
PAH Mass Removals (Grams/Day)

      IT Project No. 408250
Date
10/10
10/19
10/26
11/2
11/9
11/16
11/22
12/2
12/8
12/15
12/30
1/5
1/26
2/2
2/16
3/2
3/9
3/16
3/30
4/13
4/19
Total Operational
Period Average
Average Prior to
Average Following
3/2
1 - \
Reactor 1 | Reactor 2 , Reactor 3
j
35.5 0
0 1.78
0 i 3.68
15.6 4.00
19.7 I 4.90
7.13 2.78
2.65 5.35
7.86 4.14
9.57 11.23
12.67 4.30
13.82 4.96
17.50
11.49
0
0
0
5.95
4.24
0
1.43
6.75
10.55
|
12.48 1.26 5.61
12.52
13.02
14.01
7.62
7.70
7.85
7.54
7.90
7.71
10.61 ±7.5
11.51 ±8.45
7.74 ±0.14
3.15
4.06
3.32
2^7
1.26*
0
1.75
1.47
1.38
3.21 ±2.43
3.93 ± 2.49
1.17 ±0.68
0
0
0.90
1.10
0
0.29
0.58
0.47
1.06
3.23 ± 4.83
4.29 ±5.39
0.48 ±0.39
     ± Indicates standard deviation.
               37

-------
            Table 4-6
CPAH Mass Removal (Grams/Day)

      IT Project No. 408250
Date
10/10
10/19
10/26
11/2
11/9
11/16
Reactor 1
0.44
0
0
0.65
1.61
0.18
11/22 0
12/2 1 0
12/8 2.33
12/15
12/30
J
1/5
1/26
]
2/2
2/16
3/2
3/9
3/16
3/30
0.24
4.02
0.50
0.32
1.90
0.95
3.33
1.06
0
0.44
^
4/13 . 1.31
4/19
Total Operational
Period Average
Average Prior to
3/2
Average Following
3/2
0.33
0.93 ± 1.13
0.88 ±1.14
0.63± 0.54
Reactor 2
0
0.31
0.52
0.47
0.76
0.59
0.73
0.43
1.00
0.24
0.76
0.19
0.34
0.69
0.49
0.55
0.23
0
0.34
0.29
0.25
0.44 ± 0.26
0.50 ± 0.26
0.22 ±0.13
Reactor 3
0.29
0.56
0
0
0
0.97
0.44
0.28
0.13
0
1.82
0
0
0.12
0.06
0.01
0
0.08
0.07
0.10
0.16
0.24 ± 0.43
0.29 ± 0.49
0.08 ± 0.06
     •S. Indicates standard  deviation.
               38

-------
                   Table 4-7
16-Hour Toxicity Determination Using  Tetrahymena

             IT Project No. 408250
?$amp le
Influent Feed



Reactor 1


Reactor 3


Sample Dilution
1:10
1:20
1:40
1:100
1:40
1:100
1:200
1:40
1:100
1:200
Percent Mortality
100
90
50
NOEC1
100
50
NOEC
100
50
NOEC
          -No  observable  effect concentration
                        39

-------
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                                                                                                               IKTEHHAT10HAL
                                                                                                               TtCHHOU)GY
                                                                         40

-------
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                                                                                 30E ACCtSl COvtHi Ql ©
                                                                                                 u* to IAM< oon
                                                                                          SECTION B-B
                                                               E1MCO Biolift™ Reactor
SECTION  A-A
                                                            BMCO PROCCSS  EQUTMENT COMPANY -  Salt l«K> City. Utah
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                                                Tu
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                                                        41

-------
9

8

7


6
                    FIGURE 41
                REACTOR pH DATA
4 -


3

2


1
          i
         0
 i .•
10
      20     30
Days of Operation
40
50
       •  Reactor 1
       •   Reactor 2
       ^  Reactor 3
                           42

-------
                        FIGURE  4-2


                  DISSOLVED OXYGEN DATA
   50
   40 -
I "30

a


"C
O)
ba
>s
T3
Ol
£  10 -
    0 -
      0
10     5    20    25


     Days of Operation
30    35
40
                Reactor 1

                Reactor 2

                Reactor 3
                                 43

-------
                                    FIGURE 4-3
                             REACTOR TOTAL SOLIDS
  60 -
  50
CO

140
C/D
I

-------
                                FIGURE 4-4

                        REACTOR VOLATILE SOLIDS
  30
  20
~o
CO



1
0)
Q.
  10
   0
4 Oct     1 Nov
              1
                                4 Jan 19 Jan 2 Feb







                                —•—     2

3

-------
                                 FIGURE 4-5
                         RATIO OF VOLATILE SOLIDS
                              TO TOTAL SOLIDS
 0.1
0.01
40ct
             1 Nov
2 Dec
                 Reactor 1
4 Jan      2 Feb
 Sampling Dates

 —•— Reactor 2
      46
19 Apr
                                                       Reactor 3

-------
   70 -T-
                                               4-6

                                                 IN             1
   80
   50
^**


-------
  70
  60
  50
c
m


I40
"••^
at
to 30
ts
  20
  10
                                FIGURE 4-7

                   SOLIDS DISTRIBUTION IN REACTOR 3
       10Oct
2Nov
           Sample Port 1
8 Dec    4 Jan        2 Feb


         Sampling Date



     •• Sample Port 2
2 Mar        18 Apr
                                          Sample Port 3
                                    48

-------
0
                             FIGURE 4-8
               REACTOR AMMONIA CONCENTRATION
        Reactor 1
i   i  i  i
30 Dec  19 Jan
  Sampling Date
      Reactor 2
                                          i  i  i
                                          8Feb
  i  i  i
2 Mar
  i     i  r
30 Mar  19 Apr


   Reactor 3
                                   49

-------
  500
  400
  30°
.§3

li 200
c

o
                                FIGURE 4-9

                  REACTOR PHOSPHATE CONCENTRATION
CO

E?
o
c
  100
    0
      6Oct
2Nov 22Nov
              Reactor 1
                             15-Dec     19 Jan  8 Feb  2 Marie Mar


                                Sampling Date
19 Apr
                                    Reactor 2



                                     50
                                          • Reactor 3

-------
                           FIGURE 4-10
     REACTOR TOTAL HETEROTROPHIC BACTERIAL POPULATIONS
3Oct
            Reactor 1
 5 Jan     2 Feb
Sampling Dates
 —•— Reactor 2
     51
2 Mar
30 Mar   19 Apr
       Reactor 3

-------
                                   FIGURE 4-11

                         SOLID PHASE TOTAL CARBON
25000
20000
15000
10000
 5000
    0
         13
         O

         o
o
2

CM
            Reactor 1
a
OJ
O

CM
c
m
~3
to
xa
ID
y_
CM
                                   Sampling Date
(0



CM
              Reactor 2
                                       52
                    Reactor 3
                                                CO
                                                                o
                                                                CO
Q,

<

o>
                            Influent feed

-------
                                     FIGURE 4-12

                                       TOTAL

^      H
'£
CL
a.

c

s
"t.
18
u

   1500
o

tf)
CO

f
to
o
03
3
O"

^^   500
      0

t3
O
o
                   o
                   Z

                   CM
O
0>
Q-
©
U.
                                                             CO
            10
Q.

<

O>
                                     Sampling
                Reactor
                    2
               53


-------
           Figure 4-13
     % REDUCTION - REACTOR 1
            SAMPLE DATE
- PAH
CPAH
CPAH GOAL
                 54

-------
           Figure 4-14
   % REDUCTION - REACTOR 2
                          CM
_ — _f_ — ,
-CJ
tt
u.
to
m
— — i — —
*«»
so
2
*
CD
u.
m
5
I
»*-— —t— —
*«,
2
o
a
*
-— i
Q.
cn
                                             en
           SAMPLE DATE
PAH
CPAH
CPAH GOAL
                 55

-------
             Figure 4-15
    %              -           3
PAH



                 56

-------
                                Figure 4-16
                         % REDUCTION - OVERALL




z
o
p
o
Q
UJ
CC






100 -I
90
80
70 !

60 l~

50

40
30
20
o -
o
O
6
o
O
o
(O
CN
O
2
CN
o
z
o
2
(O
o
Z
CN
CM
                     PAH
T 	 ' — 1 	 '
tl O
0) at
3 0
N CO
_j__
O
«
Q
i
	 	 T^
O
O
6
SAMPLE
— — t— — i — -
c c
ra ro
-, -j
to 

GOAL
— — f-
10
2
ci


TO
2
6
ex
<
CO
                                                                              a.
                                       57

-------
                     Figure 4-17
                FLOURENE CONCENTRATIONS
u.
              DATE
DR3
• R2
• RI
S3 INFLUENT
                                   58

-------
          FIgyre 4-18

DATI
                                                              DR3
                                                              BR2
                                                              • «,:
                                                              E

-------
        Figure 4-19
    PAH CONCENTRATIONS
a
jj

"ra

JE





2
DATE
                                                        DR3



                                                        Bn2



                                                        • R1



                                                        SI INFLUENT
                     60

-------
        Figure 4-20
    CPAH CONCENTRATIONS
DATE
S; « m

-$lisStn
  en ^i tO ^- OJ J»l
  * * ri r- o ^? P
     ^ 12 52 o>
       «^5
                              R1
                            R3
                                                 DR3



                                                 • R2



                                                 • RI



                                                 d INFLUENT
                 61

-------