EPA/540/R-05/006
                                             April 2005
   Treatability Study Report of Green D
       Mountain Laboratories, Inc. s D
          Bioremediation Process D

Treatment of PCB Contaminated Soils, at
 Beede Waste  Oil/Cash  Energy Superfund
       Site, Plaistow, New Hampshire D
                         by
              Science Applications International Corporation
                    Hackensack, NJ 07601
                   Contract No. 68-C5-0036
                   Work Assignment No. 0-6

                  Technical Project Manager

                     Vicente Gallardo
             Land Remediation and Pollution Control Division
             National Risk Management Research Laboratory
                    Cincinnati, Ohio 45268
             National Risk Management Research Laboratory
                Office of Research and TableDevelopment
                U.S. Environmental Protection Agency
                    Cincinnati, Ohio 45268

-------
                                  NOTICE
This document was funded by the U.S. Environmental Protection Agency's (EPA) Superfund
Innovative Technology Evaluation (SITE) Program under Contract No. 68-C5-0036, Work
Assignment No. 0-6, to Science Applications International Corporation.  This document has
been subjected  to  EPA's  peer and administrative  reviews and  approved for
publication as an official EPA document.  Mention of trade names or commercial products
is incidental and constitutes neither endorsement nor recommendation for use.

-------
                                FOREWORD
The U.S. Environmental Protection Agency (EPA) 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 (NRMRL) is the Agency's center for
investigation of technological and management approaches for preventing and  reducing
risks from pollution that threaten human health  and the  environment. The focus of the
Laboratory's research program  is on methods and their cost-effectiveness for prevention
and control  of pollution to air, land, water, and subsurface resources; protection of water
quality in public water systems;  remediation of contaminated sites, sediments and ground
water; prevention and control of indoor air pollution; and restoration of ecosystems. NRMRL
collaborates with both public and private sector partners to foster technologies that reduce
the cost of compliance and to anticipate emerging problems. NRMRL's research  provides
solutions to  environmental problems by: developing and promoting technologies that protect
and improve the environment; advancing scientific and engineering information to support
regulatory and policy decisions; and providing the technical support  and information transfer
to ensure implementation of environmental regulations and strategies at the national, state,
and community levels.

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.
                                 Sally Guiterrez, Acting Director
                                 National Risk Management Research Laboratory
                                       in

-------
                                ABSTRACT
In 1998, under the sponsorship of the New Hampshire - Department of Environmental
Services (NHDES), Green Mountain Laboratories, Inc. (GML) and the USEPA agreed to
carry out a Superfund Innovative Technology Evaluation (SITE) project to evaluate the
effectiveness of GML's Bioremediation Process for the treatment of PCB contaminated
soils at the Beede Waste Oil/Cash Energy Superfund site in Plaistow, New Hampshire
(hereinafter referred to as the Beede site). The treatment process involved
inoculation/augmenting of the PCB contaminated soils with bulk microbial inoculum and
nutrients, and allowing the microbes to aerobically degrade the PCBs. The bulk
inoculum was produced on-site by the developer using animal feed-grade oatmeal as
the substrate, shredded pine needles that provided certain specific co-metabolite
compounds, nutrients and a proprietary consortium of microorganisms capable of
degrading the PCBs to their eventual endpoints - carbon dioxide and mineral halides.

The results of the field evaluation of the technology, which are based on the data
collected from the treatability study conducted in the third quarter of 1998,  indicate no
removal/degradation of the PCBs. In earlier laboratory tests, GML had used
concentrated pine extract to provide the co-metabolite compounds, whereas, for the field
study it used shredded pine needles.  At the end of the field treatability study, based on
its own observations and data, GML concluded that it may have inadvertently made
some fundamental errors in the production and application of the bulk inoculum.
Subsequently, the EPA SITE program and the NHDES agreed to give GML another
opportunity to demonstrate its technology's capability in degrading PCB in the Beede
site soil, but at a much smaller laboratory scale. In September 2000, GML carried out a
limited number of preliminary bench-scale tests, at the Middlebury College in
Middlebury, Vermont to reestablish the viability of its process. At the conclusion of the
bench-scale tests, GML conceded that, at best the tests were inconclusive and at worst
had failed. The project was terminated at that time.
                                      IV

-------
                        TABLE OF CONTENTSD
Section
Notice	  ii D
Foreword  	iii D
Abstract  	iv n
Tables	vi D
Acknowledgments  	  vii n

Section  1D   INTRODUCTION  	  1 D
        1.1 DSITE Program	 2 D
        1.2 D Demonstration Program  	 3D
        1.3 DSelection into the Demonstration Program	 3D
        1.4 D Points of Contact	 4 D

Section  2D   TECHNOLOGY DESCRIPTION	 6 D
        2.1 D Introduction	 6 D
        2.2D The GML Bioremediation System	 7 D
             2.2.1  Technology Description	 7 D
             2.2.2  Process Development	 7 D

Section  3 n   TREATABILITY STUDY  	 9 D
        3.1 D Treatability Study Overview  	 9 n
        3.2 D Waste/Soil Selected for Testing 	  10 D
        3.3D Objectives and Scope of the Treatability Study  	  10 n
        3.4 DExperimental Design	  12 D
        3.5 DField Operations	  14 n
             3.5.1  Plot Design and Construction 	  15 D
             3.5.2  Inoculum Production	  15 n
             3.5.3  Soil Preparation and Inoculation	  16 D
             3.5.4  Plot Maintenance  	  18 n
             3.5.5  Support Equipment and Facilities	  19 D
             3.5.6  Decontamination, Waste Disposal and Contingency Plan  ....  19 n
        3.6D Sample Collection and Analysis	  19 D

Section  4 n   RESULTS AND DISCUSSION 	  24 D
        4.1 D Introduction	  24 D
        4.2 DSoil Characterization	  24 D
        4.3 DPerformance Data	  24 D
             4.3.1  PCB Data for Treatments and Controls  	  24 D

Section  5 n   QUALITYASSURANCE	 34 D
        5.1 DQA Summary 	  34 D
        5.2 DData Quality Indicators 	  34 D
        5.3D Conclusions and Data Quality Limitations  	  35 D

Section  6 D   REFERENCES	  37 D

-------
                                TABLES
Table
Table 3.1-1   Summary Results of the June 1998 Bench-Scale Tests	  10 n
Table 3.4-1   Experimental Design Matrix  	  14 D

Table 4.2-1   Summary of PCB Aroclor Data for Various Treatments and Controls .  26D
Table 4.2-2   Summary of PCB Congener Data for Various Treatments and Controls 28D
Table 4.2-3   Summary of PCB Homologue Data for Various Treatments and ControlsD
              	  33 D
                                    VI

-------
                         ACKNOWLEDGMENTSD

This report was prepared under the direction of Vicente Gallardo, Technical Project
Manager, U.S. Environmental Protection Agency (EPA), National Risk Management
Research Laboratory (NRMRL), Cincinnati, Ohio. A key contributor included Dr.
Anthony Rutkowski (formerly with Green Mountain Laboratories, Inc.). EPA NRMRL
peer review of this report was conducted by Dr. Ronald F. Lewis and  Dr. Ronald
Herrmann. New Hampshire Department of Environmental Services (NHDES) peer
review was conducted by Robert P. Minicucci, II.

This report was prepared for EPA's Superfund Innovative Technology Evaluation (SITE)
program by the Environmental Technology  Division of Science Applications
International Corporation (SAIC) in Hackensack, New Jersey under Contract No. 68-C5-
0036,  WA# 0-6.  Laboratory analyses described in this report were performed by
Northeast Analytical Environmental Lab Services (Schenectady, NY)  and Alta Analytical
Laboratory, Inc.  (El Dorado Hills, CA).
                                     Vll

-------
                                    Section  1.0
                                 INTRODUCTION

In 1980, the U.S. Congress passed the Comprehensive Environmental Response, Compensation,
and Liability Act (CERCLA), also known as Superfund, committed to protecting human health and
the  environment  from uncontrolled  hazardous waste sites.   CERCLA was amended  by  the
Superfund Amendments and Reauthorization Act (SARA) in 1986. These amendments emphasize
the  long term effectiveness and permanence of remedies at Superfund sites.  SARA mandates
implementing permanent solutions and using alternate treatment technologies or resource recovery
technologies, to the maximum extent possible, to clean up hazardous waste sites.

State and Federal agencies, as well as private parties, are now exploring a growing number of
innovative technologies for treating hazardous waste. The sites on the National Priorities List total
more than 1,200  and comprise a broad spectrum of physical, chemical,  and environmental
conditions requiring varying types of remediation.  The U.S.  Environmental Protection Agency
(EPA) has focused on policy, technical, and informational issues related to exploring and applying
new remediation technologies  applicable to Superfund  sites.  One such initiative  is EPA's
Superfund Innovative Technology Evaluation (SITE) Program, which was established to accelerate
development, demonstration, and use of innovative  technologies for site cleanups.

In  1998,  a  bioremediation  process  developed by  Green Mountain  Laboratories,  Inc.  (GML),
Middlesex, Vermont was field-tested under the SITE program.  Through  its research in  the
laboratory and some limited involvement in PCB remediation projects, GML believed that it had put
together a process combining engineering, chemistry, and microbiology to remediate PCBs in soil.
GML believed that its unique on-site  processing  technique  using indigenous microbes,  co-
metabolites and nutrients could drive the bioremediation of PCBs  to their ultimate  endpoint of
carbon dioxide and mineral halides.

A pilot-scale treatability study of the GML PCB Bioremediation process was performed at the Beede
Waste Oil/Cash Energy site (hereafter referred to as Beede site) in Plaistow, New Hampshire, over
an eight week period,  from August through October 1998.

The results of the field study indicated that the GML process was unsuccessful in  degrading the
PCBs.  In earlier laboratory tests, GML had used a concentrated extract of pine needles to provide
the  co-metabolite compounds to facilitate PCB degradation, whereas for the field  study,  it used
shredded pine needles.  At the end of the field study, based on its own observations and data GML
concluded that it may have inadvertently made a fundamental error in the production of the bulk
inoculum by using the shredded pine needles instead of the concentrated extract. Consequently,
the  EPA SITE program and the NHDES agreed to give GML another opportunity to demonstrate
the technology's capability in degrading PCBs in the Beede site soil. However, GML was asked
to first reestablish the viability of the process on a much smaller laboratory scale before conducting
another field demonstration. In September 2000, GML conducted a limited number of preliminary
bench-scale tests at the Middlebury College in Middlebury, Vermont to examine its process and the
processing protocols.  At the conclusion  of the bench-scale tests, GML conceded that, at best the
tests were inconclusive and at worst had  failed to establish the capability of its PCB bioremediation
process.  The project was terminated at that time.

This treatability study report, organized into six sections, describes the GML technology, provides

-------
information pertaining to the staging of the treatability tests, and analyzes data from the field trial.
Section 1 presents background on the SITE program, the selection of the GML technology into the
Demonstration Program, and lists  points of contact for GML,  NHDES and the SITE program.
Section 2 describes the fundamentals of the GML PCB bioremediation process, and the research
and development work that led to the technology's current design. Section  3 summarizes the
treatability study from the planning stage through the field trial.  Section 4 analyzes the data and
discusses the results.  Section 5 reviews quality assurance/quality control (QA/QC) issues.
Section 6 lists technical references used in developing this report.

Although the GML technology did not meet its treatment objectives, publishing the results of the
field treatability study is still worthwhile. By carefully documenting the experimental design of the
project and describing its results,  researchers can advance the  technology by exploring new
approaches.

This  report represents the only published EPA document resulting  from this SITE Program-
sponsored project.

1.1      SITE Program

The SITE Program is a  formal program established by the EPA's Office of Solid Waste and
Emergency Response (OSWER) and Office of Research and Development (ORD) in response to
the Superfund Amendments and  Reauthorization  Act  of 1986 (SARA).   The SITE Program
promotes the development, demonstration, and use of new or innovative technologies to clean up
Superfund sites across the country.

The SITE Program's primary purpose is to maximize the use of alternatives in cleaning hazardous
waste sites by encouraging the development and demonstration of new, innovative treatment and
monitoring technologies.  It consists of three major elements:

        the Demonstration Program,

        the Consortium for Site Characterization Technologies (CSCT), and

        the Technology Transfer Program.

The objective of the Demonstration Program is to develop reliable performance and cost data on
innovative  technologies  so that potential users  can  assess the  technology's site-specific
applicability. Technologies evaluated are either available commercially or close to being available
for full-scale remediation  of Superfund sites.  SITE demonstrations  usually are  conducted at
hazardous waste sites under conditions that closely simulate full-scale remediation conditions, thus
assuring the usefulness and reliability of the information collected. Data  collected are used to
assess: (1) the performance of the technology; (2) the potential need for pre- and post-treatment
of wastes; (3) potential operating problems; and (4) the approximate costs. The demonstration also
provides opportunities to evaluate the long term risks and limitations of a technology.

Existing and  new technologies and  test  procedures  that improve  field monitoring and site
characterizations are explored in the CSCT Program. New monitoring  technologies, or analytical
methods that provide faster, more cost-effective contamination and  site assessment data are
supported by this program. The CSCT Program also  formulates the protocols  and standard

-------
operating procedures for demonstration methods and equipment.

The Technology Transfer Program disseminates technical information on innovative technologies
in the Demonstration and CSCT Programs through various activities. These activities increase
awareness and promote the use of innovative technologies for assessment and remediation at
Superfund sites. The goal of technology transfer activities is to develop interactive communication
among individuals requiring up-to-date technical information.
1.2     The SITE Demonstration Program and Reports

For the  first ten years in the history of the SITE program,  technologies had been selected for
evaluation through annual requests for proposals. EPA reviewed proposals to determine the
technologies with promise for use at hazardous waste sites. Several technologies also entered the
program from current Superfund projects, in which innovative techniques of broad interest were
identified under the program.

In  1997  the program shifted from a technology driven focus to a more integrated  approach driven
by the needs of the hazardous waste remediation community. The SITE program now annually
solicits applications for participation in the Demonstration program from parties responsible for
clean up operations at hazardous waste  sites. A team  of stakeholders led by SITE program
personnel will select  sites  and work  with site representatives  in  bringing  technologies for
demonstration to their respective sites.

Once the EPA has accepted an application, cooperative arrangements are established among EPA,
the developer, and the stakeholders to set forth responsibilities for conducting the demonstration
and evaluating the technology. Developers are responsible for operating their innovative systems
at a selected site, and are expected to pay the costs to transport equipment to the site, operate the
equipment on site  during the demonstration, and remove the equipment from  the site.  EPA is
responsible for project planning, sampling and analysis,  quality assurance and quality control,
preparing reports, and disseminating information. Typically, results of Demonstration Projects are
published in three documents: the SITE Demonstration Bulletin, the Technology Capsule, and the
Innovative Technology Evaluation Report( ITER). The Bulletin describes the  technology and
provides preliminary results  of the field demonstration. The Technology Capsule provides more
detailed information about  the technology and  emphasizes key results of the SITE field
demonstration. An additional report, the Technology  Evaluation Report (TER), is available by
request  only.  The TER contains a comprehensive presentation of the data collected during the
demonstration and provides  a detailed quality assurance review of the data.

However, with the GML study, the technology did not advance to a full Demonstration, thus only
a treatability study report will be published.

1.3     Selection into the Demonstration Program

In the past, technologies were selected for the Demonstration  Program from a pool of responses
to  SITE'S annual request for proposals (RFP). EPA reviewed proposals to search for innovative
technologies that offered either reduced risk or cost or provided a treatment solution where none
had existed previously. In 1997, the program shifted from a technology-driven  focus to  a more
integrated approach shaped by the needs of the hazardous waste remediation community. The

-------
annual RFP was discontinued, and instead a team of stakeholders matches technologies with a
particular site that has been selected for study by the SITE program. The stakeholders and EPA
solicit and evaluate proposals from technology developers interested in working at the chosen site.

In its "Host Site Application (HSA) to the SITE Demonstration Program" the State of New Hampshire -
Department of Environmental  Services (NHDES), on February 26,  1998,  identified one such
innovative technology. The technology identified is a bioremediation process for the treatment of soils
contaminated with polychlorinated biphenyls (PCBs) developed by Green Mountain Laboratories, Inc.
of Middlesex, Vermont (hereinafter also referred to as the "Developer"). In the HSA, the NHDES also
identified a candidate site for demonstrating and evaluating the GML technology; the Beede Waste
Oil/Cash  Energy Superfund Site in Plaistow, New Hampshire (hereinafter also  referred to as the
Beede site).

1.4 Points of Contact

Additional information on the  GML Technology, the Beede site and the SITE Program can  be
obtained from the following sources:

The GML PCB Bioremediation Technology:

Raul Sanchez
Green Mountain Laboratories,  Inc.
27 Cross Road
Middlesex, Vermont 05602
Telephone:    (802)223-1468
Fax:          (802) 223-8688
Email:       gml@together.net

The Beede Site:

Robert P. Minicucci, II
Innovative Technology Coordinator
New Hampshire Department of Environmental Services (NHDES)
P.O. Box 95
6 Hazen Drive
Concord, New Hampshire 03301
Telephone:    (603)271-2941
Fax:          (603)271-2456
Email:       rminicucci@des.state.nh. us

The SITE Program:

Annette M. Gatchett, Acting Director
Land Remediation and Pollution Control Division
U.S. Environmental Protection Agency
26 West Martin Luther King Drive
Cincinnati, OH  45268
Telephone:    (513)569-7697
Fax:          (513)569-7620
Email:       gatchett.annette@epa.gov

-------
Vicente Gallardo
EPA SITE Technical Project Manager
U.S. Environmental Protection Agency
26 West Martin Luther King Drive
Cincinnati, OH  45268
Telephone:   (513)569-7176
Fax:         (513)569-7620
Email:        gallardo.vincente@epa.gov

Information on  the SITE  program is  available  through  the  following  on-line information
clearinghouses:

        • D   The  SITE Home  Page  (www.epa.gov/ORD/SITE)  provides  general  program
             information, current project status, technology documents, and access to other
             remediation home. Note: URL is case sensitive.

        • D   The  OSWER  CLU-ln electronic  bulletin board (http://www.clu-in.org)  contains
             information on the status of SITE technology demonstrations.

Technical reports may be obtained by writing to USEPA/NSCEP, P.O. Box 42419, Cincinnati, Ohio
45242-2419, or by calling 800-490-9198.

-------
                                    Section 2.0
                         TECHNOLOGY DESCRIPTION
2.1      Introduction
Polychlorinated biphenyls (PCBs) are mixtures of synthetic organic chemicals that possess useful
industrial characteristics. They are chemically stable, have low vapor pressure, low flammability,
high heat capacity, low electrical conductivity, and high dielectric constant.   Based on these
properties commercial PCB mixtures were  used  in many industrial applications, especially in
capacitors, transformers, and other  electrical equipment. They were also used, but to a lesser
extent, as plasticizers, hydraulic fluids and lubricants, carbonless copy paper, heat-transfer fluids
and petroleum additives. The unique chemical properties, also contribute to the  persistence of
PCBs after they are released into the  environment.  Evidence that PCBs persist in the environment
and  may  cause environmental and health hazards  stopped  the domestic manufacture  of
commercial mixtures in  1977.  In 1976, US Congress enacted the Toxic Substance Control Act
(TSCA), which directed the EPA to control the manufacture, processing, distribution, use, disposal,
and labeling of PCBs. Regulations under TSCA govern all forms and combinations of chemicals
which contain the biphenyl molecule with one or more chlorine atom substitutions. They also apply
not only to the PCB chemicals themselves, but to items and materials which have been in contact
with PCBs.

In the environment, PCBs also occur as mixtures of congeners, but their composition may differ
from the commercial mixtures. After release into the environment, the composition of PCB mixture
can  change   over  time   through  partitioning,  chemical  transformation,  and  preferential
bioaccumulation. PCBs adsorb to organic materials, sediments, and soils. PCBs are widespread
in the environment, and humans can be exposed through multiple pathways.  Levels in air, water,
sediment, soil, and foods vary over several orders of magnitude, often depending on proximity to
a source of release into the environment.

Based on the  1990 EPA Superfund guidelines, the  NHDES has adopted the stringent soil cleanup
standard of total PCBs < 1 mg/kg or ppm for residential areas.  In the NHDES policy, the S-1  and
S-2 standard  is 1  ppm, while the S-3 standard is 2 ppm.  The S-1 and S-2 standards apply to
situations where soil is more accessible, whereas the S-3 standard is for fairly inaccessible soil.
The current cleanup goals for PCB Superfund sites in EPA Region 1 are determined on a case-by-
case basis and determined  by the risks posed by each site.

In  1998, from August through October,  over an eight-week period, a pilot-scale field treatability
study of the GML technology was conducted at the Beede site in Plaistow, New Hampshire. The
primary purpose of the treatability study was to provide an initial assessment of the effectiveness
of the GML process in achieving or approaching either of these goals (the DES S1/S2 standards)
under actual and/or simulated field conditions (soil character,  other contaminants,  weather, etc.).

Prior to entering the SITE program, GML conducted laboratory scale experiments on Beede soils
using a unique combination of indigenous microbes, co-metabolites and nutrients. Based on the
findings of these preliminary laboratory experiments, GML believed that it had developed the know-
how that could be used to successfully degrade the full range of PCB congeners to innocuous final
products, particularly carbon dioxide and mineral halides. GML claimed that it had designed its
proprietary microbial consortium so  that it would  be able to successively degrade the  various

-------
intermediates that may be produced as well as the PCB congeners that may be present at a site.
GML also claimed that the technology was designed to degrade PCBs in the presence of other
contaminants such as oils (petroleum hydrocarbons) and heavy metals. The Beede site provided
a unique opportunity to evaluate all these aspects of the process.

Information from the treatability study was to be used to design a larger scale system for treating
a larger range and quantity of the contaminated soil at the site. I nformation from the study was also
used to provide initial assessment of the applicability of GML bioremediation technology to other
sites with similar waste characteristics.
2.2     The GML Bioremediation System

2.2.1    Technology Description

The GML PCB Bioremediation process involves bioaugmentation of the PCB contaminated soil with
a customized microbial inoculum and a proprietary nutrient formulation.  The bulk inoculum is built
on food or animal grade grain (in the case of this study GML used oatmeal) that serves as the
substrate  as well  as  the  food source for the proprietary consortium  of  PCB degrading
microorganisms. The GML designers of the inoculum have indicated that in addition to the oatmeal
and the microbial consortium, the extract of pine (specifically Spruce pine) needles forms a key
constituent of the  inoculum.   According to  GML,  the  Spruce  pine extract  supplies the
microorganisms with terpenes (naturally occurring  compounds  found in Spruce pine  needles)
which serve as co-metabolites for the PCB degrading microbes. As per GML, as the microbes get
acclimated towards the terpenes and other pine constituents they develop a greater affinity for PCB
molecules, and thereby end up consuming/degrading the PCBs.

2.2.2    Process Development - As Described by GML

As originally conceived, this project was intended to provide an inexpensive, natural and efficient
method forthebiodegradation of Polychlorinated Biphenyls(PCB).  Of particular interest were those
of higher molecular weight, such  as Aroclor,  1254 and 1260 that had proven recalcitrant  to
biodegradation.  This recalcitrance is based on  the difficulty of dechlorinating highly substituted
biphenyls and by the accumulation of by-products toxic to  organisms capable of initiating the
dehalogination and cleavage of the biphenyl rings. Only naturally occurring organisms were to be
employed and ideally the process was to be aerobic, as anaerobic conditions are logistically harder
to maintain.  It was also realized that no one organism was likely  to carry out all necessary steps
in the process, so a synergistic consortium of microbes was sought.

Isolation of microorganisms capable of tolerating the presence of and ultimately degrading PCBs
was achieved through conventional methods. Coverslips coated with either Aroclor 1254 or 1260
were  inserted into  a dish containing soil known to be  contaminated  with Aroclors.  A broth
containing mineral salts, ammonium and trace metals intended to make up for deficiencies in the
soil was added. Coverslips were then withdrawn at intervals; washed to remove loose debris and
transferred to vials  containing mineral salts, ammonium, trace  metals and an Aroclor.  Samples
were transferred from vials demonstrating growth to a second vial, again containing mineral salts,
ammonium, trace metals and Aroclor.  Growth from this second set of vials was transferred to a
third set from which regular passages were  maintained in the same fashion as eukaryotic cell
cultures, as well as being streaked onto an  assortment of agar based solid media to facilitate
isolation of the organisms.  In this way, two aerobic,  gram-negative rods were isolated which

-------
demonstrated the potential to exist in tandem in or on minimal media with Aroclor 1254, 1260, or
a combination of both as sole carbon source. It was also determined that one of these organisms
was a methylotroph, as survival in Aroclor was aided by the addition of small amounts of methanol
or formaldehyde.

After the initial isolation was achieved, the microbial pair was screened for its ability to degrade
Aroclor 1254 and 1260 in aerobic culture. As no standard chromatographic method existed which
could simultaneously separate PCBs from the major expected by-products, chlorocatechols and
chlorobenzoates, a non-traditional  approach was adopted. Thin layer chromatography, utilizing
silica  gel as the stationary phase and a water, methanol and acetic acid mobile phase proved
capable of clearly separating all target compounds from chloroform extracts of test cultures. Target
bands were plainly visible under ultraviolet light, and test cultures routinely exhibited evidence of
the  generation of the expected by-product not seen in uninoculated or killed control cultures.

Since the degradation of PCBs is stimulated through the use of a co-metabolite a search was
made for a suitable structural analog. (In this application, GML defines a co-metabolite as a
compound similar in structure but more easily degraded than the PCBs)  The compound most
commonly used experimentally,  biphenyl, was eliminated since  it is considered  a  hazard.  The
terpenes, a class of isoprenoid compounds common in the oils and waxes of plants, was selected,
and crude extracts of balsam fir, pine and various herbs were tried. Balsam fir needles were settled
upon as a source, being plentiful and easily  available.

After selecting the organisms, media and a co-metabolite, a  method of delivery to the soil was
investigated. The delivery mechanism needed to  be biodegradable, absorbent and economically
feasible.  Ease of storage, handling and transport were also considered.  A process was developed
in which oatmeal was saturated with mineral salt broth and an extract of balsam fir.  This mixture,
when dried and ground, resembled coarse sand. When needed, an appropriate amount could be
inoculated, allowed to grow for 72 hours, fed with  MSB and mixed with the soil.  The goal was to
insure sufficient cell density within the inoculum to allow the introduced cells to grow  and thrive
when introduced into soil. This amendment could be added to the soil and land farmed for aeration,
and sprayed with MSB and fir extract until PCB levels in the target soil fell to acceptable levels.

-------
                                    Section 3.0
                             TREATABILITY STUDY
3.1      Treatability Study Overview

Based on the 1990 EPA Superfund guidelines the NHDES has adopted the soil cleanup standard
of Total PCBs  <  1 mg/kg or ppm for residential areas.  In the  NHDES policy, the S-1 and S-2
standard is 1 ppm, while the S-3 standard is 2 ppm. The S-1 and S-2 standards apply to situations
where soil is more accessible, whereas the S-3 standard is for fairly inaccessible soil. The current
cleanup goals for PCB Superfund sites in EPA Region 1 are determined on a case-by-case basis
and determined by the risks posed by each site.

According to GML, priorto entering the SITE Program, they performed laboratory scale experiments
with the unique combination of indigenous microbes,  co-metabolites, nutrients and processing
techniques that successfully degraded the full range of PCB congeners to innocuous final products.
GML designed its microbial consortium so that it was  able to successively degrade the various
intermediates that may be produced  as well as the PCB congeners that may be present at a site.
GML claimed that the technology was particularly  designed to degrade PCBs in the presence of
other contaminants such as oils  (hydrocarbons) and heavy metals.  The  Beede site provided a
unique opportunity to evaluate all these aspects of the  process.

Thus,  the primary purpose of this treatability study was to determine how  effective the GML
bioremediation process was in achieving or approaching either of the NHDES treatment goals
under actual and/or simulated field conditions (soil character, other contaminants, weather, etc.).

If successful, information generated from this treatability study was to be used to design a larger
scale  system for treating  a larger range and  quantity of the contaminated soils at the site.
Information from this study was to provide initial assessment  of the  applicability  of  GML's
bioremediation technologies at other sites that may contain similar waste constituents.

In early June of 1998, GML conducted an experiment with six test plots at its Middlesex, Vermont
facility on PCB-contaminated soils from the Beede site. Two soils, one from location S-109 and the
other from S-43 were used. [Note: Locations S-109 and S-43 refer to the general locations where
soil samples S-109 and S-43 had  been collected,  respectively,  during an earlier  remedial
investigation study at the Beede site.] Each soil was thoroughly homogenized and split into three
parts.  For each type of soil, one part was set aside as untreated control, while the other two were
subjected to two different treatments (in terms of the consortium selected). At the end of two weeks
of treatment the four treated and  two control samples were analyzed for PCBs. Results of these
analyses are presented in Table 3.1-1.  Although limited, these results confirmed for GML that
some extent of PCB degradation was achieved. It also suggested that high concentrations of total
petroleum hydrocarbons (TPHs) may have interfered or retarded PCB degradation. Therefore, for
the subsequent  treatability study GML selected a consortium based  on  these results that
performed best with high and low TPH soils and extended the treatment period to eight weeks.

-------
             Table 3.1-1   Summary Results of the June 1998 Bench-Scale Tests

Experi-
ment
#
1
2
3
4
5
6
Treatment
or
Control
Treatment
Consortium
1
Treatment
Consortium
1
Treatment
Consortium
2
Treatment
Consortium
2
Control
Control
Soil
Source
Location
S-1 09
S-43
S-1 09
S-43
S-1 09
S-43
Reference Data
Concentrations based
on Remedial
Investigation
Total
Aroclors
(mg/kg)
250
260
250
260
250
260
TPHs
(mg/kg)
8800
600
8800
600
8800
600
June 1998 Bench-Scale Test Data
PCB Concentrations for
Bench-Scale Test Samples
(Post Treatment Experiments
1-4)
Total
Aroclors
(mg/kg)
234
45.0
376
49.6
373
119
Total
Congeners
(mg/kg)
306
48.8
489
56.3
508
140
Percent
Reductions in
Total Aroclors
based on Control
Soil
concentrations
37.3
62.2
-0.8
58.3


3.2     Waste/Soil Selected for Testing

Two locations on the Beede site with relatively high PCB concentrations (i.e. >  100 mg/kg total
PCBs) were selected based on previous analytical results.  The selection of  the  two source
locations also depended on the concentrations of other relevant contaminants (TPHs and metals),
and on availability and accessibility of the soil.  Based on initial Rl data provided by NHDES, the
two candidate sources were identified as the locations S-109 and S-43, which had shown total PCB
concentrations of 250 and 260 mg/kg, respectively, in the surface soil. Based on the needs of the
experimental design (i.e., the various treatment and control plots) discussed in Section 3.4,  it was
estimated that about 10 cubicyards of PCB-contaminated soil (excluding gross debris and material
larger than 1 inch in size) would be needed to stage the treatability study; approximately 7 cu. yd.
from location S-109 and 3 cu. yd. from location S-43.
3.3     Objectives and Scope of the Treatability Study

The primary objectives of this field treatability study were (1)  to establish the applicability of the
GML treatment process to the Beede site, and (2) to determine its effectiveness in biodegrading
the primary target contaminants, PCBs, in  soil  which also contained lead and other organic
contaminants including TPHs.
                                          10

-------
For the treatability study, GML proposed to set up and test a series of treatments and controls for
a period of eight weeks using PCB-contaminated soil from two distinct locations on the Beede site.
Details of the  experimental design are discussed in Section 3.4.  The data and information
generated from this study was to provide a better understanding of the treatment process and allow
the developer to identify the candidate treatments and controls that would be used subsequently
in a pilot-scale demonstration study. The data from the treatability study was to allow the developer
to determine the optimal treatment conditions and parameters, for customizing the treatment to the
Beede site soil. In addition, the data generated through this treatability study would enable the
developer to clearly define the primary and secondary objectives for the demonstration study and
also allow the SITE Program to develop a comprehensive site-specific demonstration test plan and
a Quality Assurance Project Plan (QAPP).

The objectives for the treatability study were:

1. D     To determine if the GML process could work under field conditions to degrade total PCBs
        from an initial concentration of 100 mg/kg or more to less than 1 mg/kg of total PCBs in
        soil, using the congener specific EPA Method 8082 or its equivalent. To accomplish this
        objective,  for each plot, the  pre- and  post-treatment average  total PCB  Aroclor
        concentrations was to be computed (along with their corresponding upper and lower 90%
        confidence limits) and reported along with the corresponding congener specific total PCB
        data.  In addition, the average percent reduction in total PCBs was to be determined for
        each plot (i.e., treatment or control) based on the baseline (Day 0) and the final (Day 56)
        total PCB Aroclor concentrations.

2.D     To estimate the PCB degradation rates for the various treatments and control by plotting
        the average total PCB Aroclor and the congener specific total PCB concentrations against
        the  treatment time. This analysis was to provide the basis for selecting the candidate
        treatments and controls for the demonstration tests. In addition, it was to provide an initial
        estimate of the treatment duration for the demonstration.

3.D     To determine the impact of TPH concentrations on the process' ability to degrade PCBs
        in soil.  This was to be accomplished by comparing the overall reductions as well as the
        degradation rates (i.e., the concentration versus time profiles)  observed in the plots with
        identical treatments but on different soils (i.e., on S-109 with high and S-43 with low TPH
        concentrations).

4.D     To  determine the  reproducibility of the  treatment  performance  data.  This will be
        accomplished by comparing the performance data (in terms  of overall reductions and
        degradation rates) obtained from duplicated experiments.

5.D     To determine if toxic by-products are being produced as a result of PCB degradation. This
        determination was  to be based on analyses of intermediate and  post-treatment soil
        samples  for chlorobenzoates,  chlorocatechols,  vinyl chloride and other chlorinated
        compounds.

6.D     To examine the toxicological  impact of the GML treatment  on the soil. This was to be
        accomplished to a limited degree by examining toxicological  data from the analysis of the
        pre- and post-treatment soil from a given treatment plot.

7.D     To examine the nature and size of microbial populations in the  various treatments and

                                           11

-------
        controls through the course of the demonstration.  This examination was to be based on
        selected microbial analyses that would be performed on soils from the same five plots
        (treatments and controls) at the start, midpoint and end of the study.

3.4     Experimental Design

In order to determine the optimal processing parameters and conditions that could be later used
to demonstrate the effectiveness of its PCB treatment process through a pilot-scale demonstration,
GML staged a series of treatments and controls which are presented in the Experimental Design
Matrix in Table 3.4-1. The design included a total of eight distinct experiments plus two duplicates
to determine reproducibility of performance data. Thus, a total of ten ex-situ plots were set up for
this treatability study.  GML designed, constructed and maintained the treatment  plots. SAIC
provided oversight during construction.

Each ex-situ plot was approximately 4' x 3.5' in size and lined on the insides and bottom with two
layers of 10-mil (or higher) polyethylene liner to prevent  escape of contaminated leachate, if
generated.  Of the ten plots, nine were protected by a roof-like cover (in the form of a removable,
raised but slanted plywood sheet cover), and one was left exposed to the elements. As the GML
treatment process was  intended to be aerobic in nature, seven  of the ten ex-situ plots were
equipped with a passive aeration system in the form of perforated corrugated PVC piping.

Surface soils (i.e., top 12 inches) from two selected locations (primarily from the locations of Rl
samples S-109 and S-43) on the Beede site were excavated, screened, analyzed on-site (using a
gas chromatograph  (GC)  with an ECD  detector)  to determine PCB  concentrations,  and
homogenized.  The  homogenized soils were then  blended  with  GML's  proprietary inoculum
(consisting  of a substrate and a custom designed consortium  of microbes) or just the  substrate
(i.e., without the specific microbes) and placed in the appropriate treatment or control plots as per
the experimental design matrix shown in Table 3.4-1  In the non-amended control plot only the
homogenized soil (i.e., without the inoculum orthe substrate) was placed. Soil was placed (loosely
packed) in  these plots to a depth of approximately 18 inches.  Plot covers were installed in a
manner that allowed easy removal and reinstallation to facilitate sample collection.

Through the course of the study, GML routinely monitored soil moisture levels and irrigated the
plots as needed. A brief description of each  plot is presented below.

Plot 1D       Ex-situ covered (rain and direct sunlight sheltered) plot equipped with corrugated
             perforated piping that provided passive aeration and received inoculum at about 5%
             by weight of the soil within the plot.  The soil used for this experiment was obtained
             from source location S-109. It did not receive methanol as a co-metabolite.

Plot 2 D       Ex-situ covered plot with no corrugated perforated piping but received inoculum at
             about 5% by weight of the soil within the plot. The soil used for this experiment was
             obtained from source location S-109. It did not receive methanol as a co-metabolite.

Plot 3D       Ex-situ covered plot equipped  with  corrugated perforated piping that provided
             passive aeration and received uninoculated substrate at about 5% by weight of the
             soil within the plot.  The soil used  for this experiment was obtained from source
             location S-109. It did not receive methanol as a co-metabolite.

Plot4D       Ex-situ covered plot equipped  with  corrugated perforated piping that provided

                                           12

-------
             passive aeration and received inoculum at about 5% by weight of the soil within the
             plot. The soil used for this experiment was obtained from source location S-109.
             In addition a liter of methanol was poured into the corrugated piping at the start of
             the experiment and every 14 days thereafter. After adding methanol, the manifold
             was plugged for a day or two to prevent off  gassing. Methanol was added as
             thestarter source of carbon for the inoculum. It did not function as a  co-metabolite
             in the sense that it was a compound similar in structure and more easily degraded
             than the PCBs.

Plot 5D       Prepared and maintained identically as Plot 4.  The experiment in  Plot  5 was a
             duplicate to Plot 4.

Plot6D       Ex-situ  covered plot equipped with a corrugated perforated  piping  that provided
             passive aeration and received inoculum at about 5% by weight of the soil within the
             plot. The soil used for this experiment was obtained from source location S-43. In
             addition, a liter of methanol, which served as a carbon source, was poured into the
             corrugated piping at the start of the experiment and every 14 days thereafter. After
             adding methanol, the manifold was plugged for a day or two to prevent off gassing.

Plot 7D       Prepared and maintained identically as Plot 6.  The experiment in  Plot  7 was a
             duplicate to Plot 6.

Plot 8D       Ex-situ  covered plot equipped with corrugated  perforated piping  that provided
             passive aeration but did not receive inoculum. The soil used forthis experiment was
             obtained from source location S-109. In addition, no methanol was added through
             the course of the study.

PlotQD       Ex-situ  covered plot with  no corrugated perforated piping  and  did not receive
             inoculum. The soil used for this experiment was  obtained from source location S-
             109. In addition, no methanol was added through the course of the  study.

Plot 10DEx-situ  experiment in a plot with  liner but no corrugated  perforated piping for passive
        aeration, and was not covered. The S-43 soil mixed with 5% inoculum was used for this
        experiment. One liter of methanol, diluted in water, was sprayed on the plot initially and
        every 14 days thereafter.
                                          13

-------
                          Table 3.4-1  Experimental Design Matrix
Plot
No.
1
2
3
4
5
6
7
8
9
10
Experimental Parameters
Source1
Location
of Soil
Used
S-109
S-109
S-109
S-109
S-109
S-43
S-43
S-109
S-109
S-43
Plot
Type
Ex-situ
Ex-situ
Ex-situ
Ex-situ
Ex-situ
Ex-situ
Ex-situ
Ex-situ
Ex-situ
Ex-situ
Plot
Sheltered/
Covered?
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
Corrugated
perforated
piping Used?
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
No
No
Inoculum or
Substrate Appli-
cation, in % of
weight of soil
5% (inoculum)
5% (inoculum)
5%
(Non-microbial
substrate)
5% (inoculum)
5% (inoculum)
5% (inoculum)
5% (inoculum)
0
0
5% (inoculum)
Methanol2
Application
No
No
No
Yes
Yes
Yes
Yes
No
No
Yes
Notes:
        Source location S-109 is known to contain higher levels of TPH (on the order of 8,800 ppm), and source
        location S-43 is known to contain lower levels TPH (on the order of 600 ppm).
        Methanol was applied to ex-situ plots 4, 5, 6 & 7 by pouring one liter into the piping manifold on day 0 and every
        14 days thereafter. Methanol was applied to Plot #10 by diluting one liter with a few gallons of water and then
        spraying it over the soil.
3.5D    Field Operations

The major components involved in the staging of this treatability study were as follows:
•D      design and construction of treatment and control plots in which the defined experiments
        were carried out. Subsection 3.5.1 describes the plot design and its construction.
•D      production of the bulk inoculum.  Subsection 3.5.2 describes the  procedures that GML
        implemented to produce the inoculum on-site.
•D      prescreening sampling, analysis and preparation of the PCB  contaminated test  soils,
        blending with  bulk inoculum  or substrate and placement in the respective treatment or
        control plots. Subsection 3.5.3 discusses the soil presampling and analysis, preparation
        and  inoculation/blending procedures used.
•D      plot maintenance. Subsection 3.5.4 describes the manner in which GML maintained the
        experimental plots through the course of the study.
•D      equipment and facilities needed to support the treatability study, which are discussed in
                                            14

-------
        Subsection 3.5.5.
•D      decontamination and waste disposal activities which are discussed in Subsection 3.5.6,
        and
•D      collection  and  analysis of representative samples and collection  of physical  and
        operational data.  Section 3.6 presents a  discussion on sample/data  collection  and
        analysis.
3.5.1D  Plot Design and Construction

The ten ex-situ treatment and control plots were staged on top of the paved area immediately
adjacent to the main building (the enclosed warehouse type structure) near the main entrance, on
the Beede site.  The following procedures was used to construct the ex-situ plots.

On the paved surface two layers of 10-mil (or thicker) polyethylene liner was laid out over an area
of about 15 feet by 60 feet.  The ex-situ plots were constructed using five discarded but clean
chipboard (wooden) crates that were available at the Beede site.  Each wooden crate was about
4 feet wide, 3 feet tall and 6.7 feet  long.  First the top cover of each crate was removed. Next,
wooden (chipboard) planks were used to divide each of the five 6.7-foot long open boxes into two
compartments, each approximately 4 feet by 3.3 feet, thus creating a total of ten experimental
chambers.  The  inside surfaces of each chamber were then  secured with two layers of 10-mil
plastic.  In each plot that was designated to receive passive aeration two lengths (approximately
12 feet) of 4 inch diameter corrugated perforated pipe were laid  across the plot in a manner that
formed a "U" shape, with the two ends of the "U" raised enough so as not to be buried under the
soil that was later placed over the pipe. The bottom of the "U"  rested on top of the plastic liner at
the bottom of the plot.  To facilitate free cross-ventilation, multiple rectangular openings were cut
about 20 inches above the plot bottom (i.e., 2  inches above the top of the soil surface).  For plots
with the passive aeration system, the top ends of the "U-shaped" perforated pipe were allowed to
stick out of the rectangular openings in the side of the plots to facilitate improved air exchange.  Plot
#10 (which was to be left exposed to precipitation)  had a solid 4  inch diameter PVC pipe open at
the top and slotted and covered on the bottom with  perforated landscape material and placed
vertically in the lower corner so that it could be used to remove leachate, if necessary. A sloped
cover made from chipboard and  lumber was constructed for each box to shed rainfall for all plots
except for #10 which was left uncovered.  The original design which called for greenhouse type
covers made with polyethylene sheeting was redesigned using solid chipboard which made for a
stronger roof structure. This redesign, however, resulted in less solar heat gain over the duration
of the test. The boxes, which were not originally designed to hold the pressure of soil and water,
were reinforced by wrapping each box with two steel bands.  Each box was then placed on two
pallets and placed on two layers of  10 mil polyethylene sheeting.
3.5.2D  Inoculum Production

Based on information from GML, at its Middlesex, Vermont facility, GMLtooka literof its proprietary
nutrient fortified suspension and inoculated it with a few microliters of its customized consortium
of microbes. After allowing the one liter suspension to incubate for about 48 hours,  it was further
split into six to eight equal parts and used in turn to inoculate six to eight five-gallon carboys filled
with the same nutrient fortified suspension. After allowing the suspensions to incubate for about
48 hours, the 30 or 40  gallons of inoculum suspension were ready to be transported to the Beede
site. The carboys were transported to the site in a refrigerated truck by road.

                                           15

-------
On August 8, 1998, personnel from Green Mountain Labs began the preparation of the microbial
substrate. Two 150 gallon open top livestock watering basins were used to prepare the substrate.
The key  ingredients were rolled oats, ground pine needles,  nutrients and bacterial culture.  A
perforated garden hose that was placed in the bottom of each tub and covered with perforated
landscape fabric was attached to an air compressor to serve as an aerator to help maintain aerobic
conditions in the substrate mixture.  Next,  both tubs were filled with a total of 700 pounds of dry
rolled oats and wetted with 30-40 gallons of water to which nutrients including ferric chloride had
been  added.  The two basins  were  inoculated with a solution  of the microbial  consortia at
approximately 16:00 on Augusts, 1998.  On August 9, approximately 600 pounds of pine needles
were ground up using a bagging  lawn mower and were then mixed into the inoculated wetted oats
in the two basins using hand held garden hoes.  The pine needles had been sitting in closed trash
cans  in the sun for several days and had begun to  compost.  The mixture was covered with
plywood.  Observation of the emergence of the bubbles at the  substrate surface indicated that
aeration was occurring but the distribution throughout the basins was not uniform suggesting that
both aerobic and anaerobic regions may have existed within the basins contents.  Care was taken
to  mix the substrate thoroughly prior to adding it to the soil.

In  addition to the inoculated substrate, a separate uninoculated batch  of substrate consisting of
rolled oats, ground pine needles, and nutrients and ferric chloride solution was mixed in a plastic
trash  can on August 9 to be used in test plot #3.
3.5.3    So/7 Preparation and Inoculation

On August 3, 1998, under the supervision of SAIC field personnel, excavation of soil from the
previously identified candidate source area was initiated. The soil excavation and screening and
the sampling of the excavated soil piles was completed  on August 4, 1998. The excavation was
performed by personnel from Sanborn, Head & Associates' (engineering contractor for NHDES) a
subcontractor,  TWM.  Soil sampling was conducted by SAIC personnel. A field engineer from
Sanborn Head & Associates was also present at the site to help coordinate the soil excavation
activities.

Soil Selected for Excavation
Based on data from the Beede Waste Oil site remedial investigation two sites had been selected
to serve as the source of the  soil to be  used  in the treatability test.  These two site were each
associated with a specific soil sample. These soil samples are designated as "S-109" and "S-43."
The following is a  description of the  soil excavation and  preparation procedure.

Preparation of S-109 Soil
An area  roughly 20 ft by 20 ft (centered  on the S-109 sample location) was marked off  for
excavation.  A polyethylene liner was placed adjacent to the excavation area and a Reed shaker
screen with  one-inch openings was placed on the liner. A backhoe was then used to dig soil,
initially from within 18 inches of the surface, and dropped into the shaker. The goal was to screen
discrete batches of approximately two cubic yards of soil and then stop excavation and transport
the soil  to separate polyethylene liners laid out nearby.  The soil was dropped into separate piles
of roughly one half of the backhoe bucket per pile using the large loader bucket on the back of the
backhoe.  Due to initial communication  problems, the first batch was larger than planned and
consisted of approximately five  cubic yards  placed  in 11 piles.  A total  of  15 batches were
screened and placed  on three 20 ft by 100 ft liners.  Batches 12 through 15 were excavated from
a depth of approximately 18 to 24 inches within the same hole. The piles were then covered and

                                           16

-------
marked with stakes that identified the batch with a number starting with "1" and then a letter starting
with "A" to designate individual piles within each screened batch. A total of 62 individual piles were
prepared in this manner.

Preparation of S-43 Soil
An area  roughly 12 ft by 12 ft (centered on the  S-43 sample location)  was marked off  for
excavation. The soil was excavated, screened, and placed on a single 20 ft by 100 ft polyethylene
liner in the same manner as for the S-109 soil except that the piles were increased in size to the
full contents of  large backhoe bucket. A total of  five screened batches were processed. The
batches were numbered 16 through 20 and resulted in a total of 15 soil piles. These piles were
marked, sampled, and covered in a similar manner as the S-109 soil was.

Pre-Screening Soil Sampling
Separate soil samples were collected from each  pile. Grab samples were collected from several
locations around the surface of the soil pile using  a stainless steel spoon. The grab samples were
mixed by hand in a stainless steel bowl. A representative sample of the composited sample was
then placed  in a small plastic container for subsequent analysis using an on-site  gas
chromatograph. After the sampling was completed the soil piles were then covered with a sheet
of polyethylene to  protect the soil from rainfall.

On-Site Soil Analysis
A gas chromatograph was set up in the on-site  trailer to be used by Raul Sanchez of GML to
perform a field screening of the samples collected  from the soil piles for PCB Aroclor 1248. The
PCB Aroclor 1248 data (based on the on-site analysis) for the staged soil piles was examined by
SAIC field   staff.  Piles with the greatest concentrations of PCBs were selected  for further
processing.  Piles were selected to ensure that sufficient soil volume was available to conduct the
treatability test.

Final Soil Preparation and Inoculation
On  August 10 and 11, 1998 a field crew from TWM returned to the site to perform the final soil
handling  activities. On August 10, 1998, the selected  S-109 soil piles were picked up with the
backhoe  and placed on a polyethylene liner and were then  mixed using the small bucket of the
backhoe.  A cement mixer was placed on the liner adjacent to the soil pile and used in order to
obtain  more thorough mixing. Once the  soil was mixed with the  backhoe, the  soil was then
transferred to the cement mixer using shovels.  In  order to estimate the soil volume per mixer batch
a five gallon bucket was filled with soil and placed in the mixer. A total of eight buckets (40 gallons)
was  needed to  fill the mixer to the maximum operating volume.  In order to simplify operating
procedures it was  decided that each soil plot would receive three mixer volumes for an estimated
total volume of 120 gallons or 0.6 cubic yards. Note that a rough measurement of the  soil after
placement resulted in a volume of 150 gallons  or 0.74 cubic yards. In order to ensure that the
controls were handled in a similar manner as the other test plots, the soil was also placed in the
cement mixer and mixed prior to placement in the test plots even though no additional materials
were added. To prevent cross contamination in the mixer, the soil to be placed in the control soil
plots was processed first.  Once the two  controls,  plots #8  and #9, were  filled, soil preparation
operations ceased for the day.

On August 11,1998, soil preparation and placement in the soil plots was resumed.  No substrate
had been used to this point. At the request of Green Mountain personnel, the volume of substrate
added to each plot was set at a volume that would maximize the volume added, ensuring that
most of the prepared substrate was utilized. This was determined to be equivalent to one and one

                                          17

-------
half five-gallon buckets full per mixer batch which as described above was estimated to be 40 to
50 gallons in volume.  Using a scale, a five gallon bucket of soil was determined to weigh 66
pounds. Assuming that the substrate has a density near that of water, the substrate application rate
was estimated to be 12.7% by weight and 18.8% by volume on a wet basis. This was two to three
times the application rate of 5% that was specified in the treatability study test plan. The developer,
during setup, produced more substrate than needed and did not want to waste any.  The test plots
were filled in a sequence that minimized cross-contamination of the microbial organisms. Because
the S-43 soil was not included in any of the uninoculated plots, this soil was processed last in the
sequence.

Plot #3 was filled with soil that had been mixed with the uninoculated substrate. The procedure for
mixing the substrate was to first put 1.5 5-gallon bucket fulls of substrate into the cement mixer and
then fill the remaining volume of the mixerwith soil. The mixerwas operated long enough to ensure
that the material was  well mixed.  Observation of the soil  substrate mixture indicated that the
substrate which consisted of a sticky oat and pine needle mixture, was broken into  clumps of 1-2
inches or less in diameter. The remaining plots containing inoculated substrate and S-109 soil were
then filled in this manner using three full cement batches  per  plot.  In order to prevent  cross
contamination of plot #3 with inoculated soil, a plywood  cover with plastic sheeting draped over
the edge was placed over plot #3 when the adjacent plot was filled.

At the request of NHDES upon completion of the filling of the plots needing S-109 soil, a 55-gallon
drum was filled with some of the remaining S-109 soil and set aside for potential use by another
party (Ttanks). The remaining S-109 soil was returned to the excavation hole and the selected soil
piles from the S-43 area were transferred to the liner for  processing.  As was done with the S-109
soil the S-43 soil was first mixed using the small bucket  of the backhoe and was then mixed with
inoculated substrate in  the cement mixer and placed in the remaining soil plots. The last plot (#10)
was filled on August 10, and the plots were watered by GML staff. Shortly thereafter, one liter of
pure methanol was added to plots #4, #5, #6, #7, and #10.  For plots #4, #5, #6, and #7 one half
of each literwas poured down into each of the two corrugated vent pipes using a funnel with a three
foot tube attached to ensure that the methanol was placed near the bottom.  For plot #10 the
methanol was dribbled across the surface of the soil.
3.5.4   Plot Maintenance

Once the treatment and control plots were setup, the only maintenance required was periodic
irrigation of the soils. GML was solely responsible for this activity and irrigated the plots on an as-
needed basis at a frequency that it deemed necessary to ensure that the plots are maintained in
proper conditions. A logbook was kept by GML to document visits, observations, moisture content,
irrigation time and rate, etc.

Water Source
Water to be used for cleaning of equipment and watering of the soil plots was obtained from an on-
site potable  water well.  Water was obtained by  placing a submersible pump  in the well and
pumping the water through garden hoses into two 150 gallon basins used to prepare the inoculated
substrate.  In order to prevent cross contamination with live microorganisms the basins were first
washed and then filled will a dilute chlorine solution and left to stand overnight. The basins were
then drained, rinsed  , and then filled again with well water. After several hours they were drained
and then filled with well water and were then ready for use. Water was drawn from the basins using
a submersible sump pump placed in one of the basins. When needed, water was pumped from

                                           18

-------
the well to the basins using a submersible pump.

3.5.5D  Support Equipment and Facilities

Typical support equipment and facilities that were required during the course of the treatability study
at the Beede site were:
•D      110V electrical power supply,
•D      potable water, for plot irrigation and decontamination of earth  handling and sampling
        equipment,
•D      a  trailer, to set up a temporary on-site analytical laboratory for analysis of screened soil
        samples, as well as for the SAIC and GML field staff to conduct administrative and clerical
        activities, and to take rest breaks,
•D      portable toilet, and
•D      earth handling equipment, suchasabackhoewith a front-end loader, a mechanical shaker
        screen (like a Reed-Screen-All) and a cement mixer.

NHDES, through its on-site contractor, SHA, provided for or facilitated the  availability of support
equipment and facilities. SAIC, with the assistance of SHA, installed a pump at the deep well (6"
O.D.) existing at the site for potable water.

3.5.6D  Decontamination,  Waste Disposal and Contingency Plan

During and at the completion of the treatability study, equipment was decontaminated by washing
with water/alconox or other suitable means (e.g. steam or pressure washing) before being moved
off-site.

All investigation derived waste (IDW) such as, personal protective gear and  other solid hazardous
waste generated during the treatability study (e.g., cinder blocks, plastic liners, manifold PVC) was
placed in approved 55 gallon drums, labeled appropriately and stored on-site where indicated by
NHDES.   Similarly,  liquid wastes  (e.g., washwater, methanol and hexane  used in  sample
preparation) were placed in separate 55 gallon drums, labeled appropriately, and staged as
directed by NHDES for ultimate disposal.

Analytical  and toxicity data generated for the soils in the various plots remaining at the end  of the
treatability study were used by  NHDES to determine the suitability of the soils for return to their
respective original source locations on the site.  If it was deemed suitable, then NHDES, through
its on-site contractor, SHA, provided for or facilitated the return of the soil to its original source area
or any other staging area on the site it determined to be appropriate.  However, if NHDES
determined that the soils were NOT suitable  for return to their respective original source locations,
then it arranged for these soils to be drummed in appropriate 55-gallon drums and staged on-site
for subsequent disposal and/or processing.

3.6     Sample Collection and Analysis

The  primary  objective of this treatability  study was  to measure  the changes  in  the  PCB
concentrations in the various treatments and controls over the course of the study by obtaining soil
samples from each plot biweekly.  SAIC was responsible for obtaining and  preparing all samples
necessary to accomplish  the objectives stated  in Section 3.3 and obtaining all the supporting
analyses.
                                          19

-------
The Test Plan (for this treatability study) dated July 1998, provided a detailed sampling plan, which
included the number and type of samples to be collected, collection frequencies, and the analyses
required. Based on the prescribed test plan, five soil grab samples from each plot per each of the
five sampling  events (fora total of 250, excluding duplicate and QC samples) were collected for
total PCB-Aroclor and other analyses. The five sampling events were conducted on Days 0, 14,
28, 42 and 56. The SITE program offered to split samples with GML during each sampling event.
However, GML requested split soil samples only from the Day 0 (orthe baseline) and Day 14 event.
The following  discussion provides a summarized account of the sample collection effort that was
undertaken for the treatability study.

Sample Sequence
Within each sample plot five grab samples were collected; one was from the center and four from
near each corner approximately  6 inches from either side and in a location that would not be
impeded by the  buried passive air  vents.   The first grab  sample was collected   from the
westernmost corner (closest to the drum storage area). The sequence then proceeded from corner
to corner in a clock-wise manner and with the fifth or last grab from the plot being from the center.

Plot Sequence
In order to minimize cross-contamination during sampling, the plots were sampled in a sequenceD
of less treatment to more treatment and secondly less soil contaminants to more soil contaminants. D
Following this rule the plots were sampled in the following order: D
Plot 9   (Control - no treatment) D
Plot 8   (Control - no treatment) D
Plot 3   (uninoculated substrate added)D
Plot 6   (inoculated substrate added,  soil with lower PCB and TPH)D
Plot 7   (inoculated substrate added,  soil with lower PCB and TPH)D
Plot 10  (inoculated substrate added,  soil with lower PCB and TPH)D
Plot 1   (inoculated substrate added,  soil with higher PCB and TPH)D
Plot 2   (inoculated substrate added,  soil with higher PCB and TPH)D
Plot 2   (inoculated substrate added,  soil with higher PCB and TPH)D
Plot 4   (inoculated substrate added,  soil with higher PCB and TPH)D
Plot 5   (inoculated substrate added,  soil with higher PCB and TPH)D

Grab Samples
Each soil grab sample was collected from the soil plots using a 2 % inch diameter coring sampler
with a cone shaped nose. The samples were collected by pushing the sampler downward until the
sampler touched the liner at the bottom of the plot or when the sampler failed to move farther with
direct pressure but no twisting. The sampler was then withdrawn  so as not to damage the liner.
Sufficient sample volume was collected from each grab except for plot 10 which  is discussed
separately below.  On subsequent sampling episodes, more than one core sample was collected
at each grab sample location when additional sample volume  was needed.

Plot #10
Due to heavy rainfall the night before collection of Day 0 samples, Plot #10 had become filled with
water that extended to the top of the soil. The consistency of the Plot #10 samples was of a slurry
and during the initial attempt to use the 2 % inch sampler with the cone tip, the sample would drop
out of the sampler as soon as the tip broke the water surface upon retrieval.  Attempts to use the
same sampler with a butterfly tip also failed. Attempts to use other diameter samplers and covering
the vent hole in the top of the samplers (at the time of retrieval) also failed. Ultimately  , the Plot #10
grab samples were collected using a stainless steel spoon by digging down no more than 6 inches.

                                         20

-------
Grab Sample Processing
The contents of the sampler were deposited in a decontaminated stainless steel bowl and was then
thoroughly mixed using a decontaminated stainless steel spoon.  Once mixed, a  portion was
transferred into sample bottles including a split for Green Mountain Lab.

Composite Sample
After all appropriate grab sample bottles were filled,  a composite sample was produced in a
decontaminated bowl by combining equal amounts of material from each of the remaining grab
samples left in their respective bowls. An  unused sample bottle served as the measuring device
to ensure equal volumes were collected from each grab. This was done starting with the bowl that
contained the least sample and if this first volume did not fill the bottle completely then the volume
collected from all of the other grabs was reduced accordingly.  This combined soil  became the
composite sample  for the plot. The composite sample was  then thoroughly mixed using a
decontaminated stainless steel spoon. The mixed composite  sample was then transferred to the
appropriate sample containers.  All  remaining sample was then returned to the  appropriate
treatment plot and the sample boreholes were physically collapsed using one of the stainless steel
spoons. All sampling equipment was decontaminated between the sampling effort for each plot.

Sampling Equipment Decontamination Procedures
In response to advice from Green Mountain Labs personnel, a modification of the first washing step
involving a dilute chlorine solution was  added to the sampling equipment decontamination
procedures to reduce contamination with any live microorganisms growing in the sampled media.
The modification involved replacing the first soak with tap water with a 15 -minute soak in a solution
of 25% chlorine bleach in tap (well) water.  In addition the first step of scraping off the equipment
with a brush was modified to a rinsing step using well water and pressurized garden sprayers. The
following sequential procedure was used to decontaminate sampling equipment and utensils:
•D      Rinse off gross soil particles with tap water using pressure sprayer,
•D      soak and scrub inside and out, for 15 minutes using a 25% chlorine bleach solution in the
        first tub,
•D      soak and scrub inside and  out, with alconox solution  in the second tub,
•D      rinse with tap water in the third tub,
•D      final rinse distilled water from a squeeze bottle,
•D      rinse with methanol from a squeeze bottle,
•D      finally, rinse with hexane from a squeeze bottle, and then,
•D      allow it to air dry on top of a flat surface covered with clean aluminum foil.

Decontamination of Sample Boring Device Between Grab Samples Within the Same Plot
Due to the limited number of sampling devices  available and time required to perform the full
decontamination procedure between each grab, it was decided that, forgrab samples within a given
soil plot, only a gross decontamination would be performed on the sample boring device used to
collect the soil sample. This decision is based on the knowledge that the contents of each soil plot
were well mixed at the beginning of the test. The gross decontamination steps used on the sample
boring device between grabs within each soil plot are as follows:
•D      Rinse off gross soil contamination using pressure sprayer and tap water,
•D      wipe the inside and outside of the boring devise with  a clean paper towel.

Water in Treatment Plots
On August 12,1998 water was observed covering approximately one half of the surface of plot #10.
Plot #10  was the only uncovered treatment plot and it had  rained approximately 2 inches (as
measured in the 5-gallon bucket left out the night before) during a heavy thunderstorm the night

                                          21

-------
before.  It was suspected that additional rainwater was driven by the wind off the side of the roof
of the adjacent plot, into plot #10. Raul Sanchez of GML was informed of this development. He
said there were no plans to remove the water in plot #10 and so it was left as is.

While sampling plot #4 water was also observed  at a depth of approximately 4 inches from the
bottom.  The observation was made by looking down into the holes left behind when the grab
sampler was removed. Raul Sanchez of GML was also informed of this observation.

Temperature Measurement
On August 12 the first treatment plot temperature measurements were made using a bi-metal type
temperature gauge in which the temperature sensitive portion is encased in a approximately 3-foot
long thin metal rod.  Two temperature measurements were made per plot. The first was made by
inserting the rod straight down into the center of the treatment plot. The gauge was allowed to
stabilize for approximately two minutes before reading. The second reading was made by inserting
the rod straight down approximately 4-6 inches in from the side of the treatment plot in the middle
of the side between the location  of the first and second grab samples.
DAY "14" SAMPLING
On Day 13, August 25, 1998, SAIC staff arrived on-site to prepare for the Day 14 sampling to be
conducted the next day.

The following day, August 26, 1998 (Day 14) soil sampling was conducted following the same
procedures established on August 12 (Day 0). As specified in the sampling plan the list of analyses
to be conducted on the composite samples was less than those analyzed for Day 0 samples. The
following  changes to the Day 0 sampling procedures were made:

•D     Due to water in the plot, the coring devise would  not retain  sufficient volume while
       sampling plot #4. Grabs 2-5 were  collected using a spoon in a similar manner as was
       done for plot #10.
•D     At the request of GML staff, the number of split samples collected was reduced from five
       to one to one per plot. This change was made to determine the treatment effectiveness,
       modify nutrient addition and because of budget constraints. The split sample was taken
       from the composite sample for each plot.

Day "28" Sampling
On Day 27, September 8, 1998, SAIC staff arrived on-site to prepare for the Day 28 sampling to
be conducted the next day.

The following day, September 9, 1998 (Day 28), soil sampling was conducted following the same
procedures established on August 12 (Day 0). As specified in the sampling plan the list of analyses
to be conducted on the composite samples was the same as those analyzed for Day 0 samples
with the exception for  metals and toxicological analyses.

So/7 Gas Sampling
Measurements of the  oxygen content of the soil gas  in the treatment plots were conducted on
September 11,1998.  The measurements were made using a thin stainless steel tube attached to
an oxygen gas meter using vinyl tubing. The stainless steel tube was equipped with a threaded rod
that could be inserted  into the tube its entire length. Appendix A presents the procedures for soil
gas sampling that were used. With the exception of the control plots #8 and #9, all of the treatment

                                         22

-------
plots exhibited reduced levels of oxygen with the lowest reading being 5.8%.

Leachate Sample Collection
Samples of the water were collected from Plots #1, #4, #5, #6, and #7.  However, the shipping
cooler was damaged during shipment and resulted in the breakage and loss of both sample bottles
from Plot #1. In between sample collection at each plot several liters of tap water were flushed
through the sample tubing to prevent gross cross contamination of the samples.

Day "56" Sampling
On  Day 55, October 6,  1998, SAIC staff arrived on-site to prepare for the Day 56 sampling to be
conducted the next day.

The following day, October 7, 1998 (Day 56), soil sampling was conducted following the same
procedures established on August 12 (Day 0). As specified in the sampling plan the list of analyses
to be conducted on the composite samples was the same as those analyzed for Day 0 samples
with the exception for metals.  Upon completion of the sampling effort the air compressor was
turned off and disconnected.
                                         23

-------
                                    Section 4.0
                            Results and Discussion
4.1      Introduction
As noted in Section 1, this project was not carried to completion (i.e., through a field demonstration)
due to strong indications that the technology did not perform effectively or as anticipated by GML,
the developer, in spite of efforts by all stakeholders to overcome the problems. Nevertheless, data
collected through the course of the eight-week field treatability study (August through October 1998)
is summarized and discussed in this section.

4.2     Soil Characterization

As discussed in Section 3, based on the soil PCB concentration data available from previously
conducted remedial investigation (Rl)atthe Beedesite, two areas that were known to be potentially
contaminated with PCBs, Rl sample locations S109 and S43, were chosen as the soil source areas
for the treatability tests. However, in order to ensure that a sufficient quantity of soil with reasonably
high PCB concentration was available from both areas, the soils from locations S109 and S43 were
carefully excavated, screened  and staged in  small piles, sampled and  analyzed  on-site, as
discussed in Section 3.

Based on the results of on-site analysis of the soil excavated from the S109 area, the PCB Aroclor
1248 concentrations ranged from 24 mg/Kg to 2,350 mg/Kg, with an average concentration of 669
mg/Kg and a median concentration of 238 mg/Kg. Of the S109 derived soil piles used to constitute
the bulk of the test soil used for the study the PCB Aroclor  1248 concentrations ranged from 154
mg/Kg to 2,350 mg/Kg, with an average concentration of 977 mg/Kg and a  median concentration
of 1,003 mg/Kg.

Based on the results of on-site analysis of the soil excavated from the S43 area, the PCB Aroclor
1248 concentrations ranged from 47 mg/Kg to 288 mg/Kg,  with an average concentration of 118
mg/Kg and a median concentration of 105 mg/Kg. Of the S43 derived soil piles used to constitute
the bulk of the test soil used for the study the PCB Aroclor 1248 concentrations ranged from 93
mg/Kg to 288 mg/Kg, with an average concentration of 154  mg/Kg and a  median concentration of
112 mg/Kg.

4.3     Performance Data

4.3.1    PCB Data for Treatments and Controls

As stated in Section 3.3, the primary objectives of this field treatability study were 1) to establish
the applicability of the GML treatment process to the Beede  site  and (2)  to determine its
effectiveness in biodegrading the primary target contaminants, PCBs, in soil which also contained
lead and other organic contaminants including TPHs.  To accomplish these objectives, a series of
experiments, as described in Section 3.4, consisting of a  few variations of GML's biotreatment
process and controls were carried out over an eight-week period from August through October of
1998 at the Beede site. The key evaluation criteria  for the these objectives was the total PCB
concentrations in  the soil that was subjected to the various treatments and controls.  Due to
budgetary constraints the  majority of the soil samples collected through the course of the study
were analyzed for the PCB Aroclor contents using the EPA Method 8082.  However, a  limited

                                          24

-------
number of composite samples from the Days 0, 28 and 56 sampling events were analyzed for
congener specific PCBs using a combination of EPA Methods 1668 and 680. Congener specific
analyses were performed to determine if biodegradation altered the Aroclor 1248 pattern.  If PCBs
were to undergo biodegradation, the original Aroclor patterns would no longer prevail and thereby
mislead one to believe that all PCBs have undergone biodegradation when in fact only a few of the
more readily degradable congeners may have biodegraded.  The rationale was, that if one or more
of the GML treatments did indeed achieve or came significantly close to achieving the treatment goal
of reducing the total PCB concentration to less than 1 mg/Kg, then the congener-specific PCB
analysis of the composite samples from the respective treatment plot would provide a more definitive
confirmation of such treatment performance.

Summary results of PCB Aroclor analysis are presented in Table 4.2-1.  These results, which are
based on at least five grab samples per plot per each of the five sampling events, clearly  indicate
that irrespective of the treatment  or control applied to the  test soil the PCB concentrations (as
Aroclor 1248)  remained unchanged after eight weeks of treatment duration. Furthermore, the PCB
concentration ranges (or the 90% confidence intervals for each plot per event) were found to be
fairly tight, thus reducing the uncertainties of the analytical findings (of the lack of performance). The
relative tightness of the confidence intervals could be attributed to fact that the target soils and the
bulk inoculum (or the non-inoculated substrate) were thoroughly homogenized prior to use in the
respective plots, thus reducing the variability in PCB concentrations across a given plot.

Summary results of the congener-specific PCB analyses are presented in Table 4.2-2  and the
homologue-specific analyses are  presented in Table 4.2-3.  These results also suggest that no
specific or noteworthy treatment effect could be attributed to any of the treatments or controls.

GML was provided with split samples from the baseline event Day 0, and Day 14, however GML
only analyzed samples  from the baseline event.  In addition, the  SITE Program made all of its
analytical results available to GML for review. Based on the results of their own sample analysis (for
PCB Aroclor) and the SITE Program results, Mr. Raul Sanchez of GML (the President) conceded
during an October 1998 teleconference with the EPA Technical Project Manager (Dr. Ronald Lewis),
the NHDES Innovative Technology Coordinator (Mr. Robert P.  Minicucci, II), and the SAIC Work
Assignment Manager (Mr. Dan Patel) that their efforts (through the field treatability tests) had failed
to demonstrate the treatment effect of their PCB Bioremediation process. Mr. Sanchez stated that
he and his  technical team (including  Dr. Tony Rutkowski, the chief microbiologist) had carefully
reviewed the treatment  procedures and methods employed prior to and during the course of the
treatability testing.   Although  they could not pin point the exact cause of the  system's non-
performance,  GML believed that the primary reason may have  been the method in which the co-
metabolite source material was applied. Mr. Sanchez elaborated that during their earlier laboratory
trials, they had use concentrated pine needle extract as the source for terpenes - the desired co-
metabolite  compounds.   However, for the  field tests, they had used  shredded pine needles.
According to Mr. Sanchez, the lack of readily available co-metabolite compounds to the treatment
system within the inoculated soil may have been a key detrimental factor.

Subsequently, the EPA SITE program and the NHDES agreed to give GML another opportunity to
demonstrate its technology's capability in degrading  PCB in  the Beede soil, but at a much smaller
laboratory scale.  In September 2000, GML carried out a limited number of preliminary bench-scale
tests, at the Middlebury College in Middlebury, Vermont to reestablish the viability of its process.
At the conclusion of the bench-scale tests, GML conceded that, at best the tests were  inconclusive
and at worst had failed.  The project was terminated at that time.

-------
Table 4.2-1    Summary of PCB Aroclor Data for the Various Treatments and Controls
PCB Aroclor 1248 Concentrations

Soil
S-109



S-109


S-109





S-109



S-109



Plnttt
Description of Experiment
Plot 1 withD
•D Inoculum at 5 % weight of soil,
•D passive aeration through
corrugated perforated piping,
•D sloped plywood cover, and
•D no methanol
Plot 2 with D
•D Inoculum at 5 % weight of soil,
•D no passive aeration through
corrugated perforated piping,
•D sloped plywood cover, and
•D no methanol
Plot 3 with
•D Non-microbial substrate at 5 %
weight of soil, D
•D passive aeration through
corrugated perforated piping,
•D sloped plywood cover, and
•D no methanol
Plot 4 with D
•D Inoculum at 5 % weight of soil,
•D passive aeration through
corrugated perforated piping,
•D sloped plywood cover, and
•D methanol
Plot 5 (a duplicate of Plot 4) with
•D Inoculum at 5 % weight of soil,
•D passive aeration through
corrugated perforated piping,
•D sloped plywood cover, and
•D methanol

StatisticD
Average
90% Confidence Interval

Median
Average
90% Confidence Interval
Median
Average D

90% Confidence Interval

Median

Average
90% Confidence Interval

Median
Average
90% Confidence Interval

Median

DayO
220
187-253

208
207
183-231
213
224

210-238

222

254
203 - 305

235
214
191 -237

205

Day 14
254
238 - 270

249
225
222 - 228
225
223

207 - 239

223

219
210-228

221
247
226 - 268

255

Day28
250
239 - 261

254
256
239 - 272
264
240

205 - 275

223

229
214-244

222
237
227 - 247

232

Day 42
267
238 - 296

269
267
193-341
239
261

210-312

230

222
213-231

224
240
227 - 253

236

Day 56
231
202 - 260

221
216
200 - 232
210
205

188-222

211

207
191 -223

205
228
220 - 236

233

-------
       Table 4.2-1    Summary of PCB Aroclor Data for the Various Treatments and Controls (continued...)
n
PCB Aroclor 1248 Concentrations

Soil
S-43


S-43


S-109


S-109


S-43




Plntfi
Description of Experiment
Plot 6 with
• Inoculum at 5 % weight of soil,
• passive aeration through
corrugated perforated piping,
• sloped plywood cover, and
• methanol
Plot 7 (a duplicate of Plot 6) with
• Inoculum at 5 % weight of soil,
• passive aeration through
corrugated perforated piping,
• sloped plywood cover, and
• methanol
Plot 8 with
• No inoculum or substrate,
• passive aeration through
corrugated perforated piping,
• sloped plywood cover, and
• no methanol
Plot 9 with
• No inoculum or substrate,
• no passive aeration through
corrugated perforated piping,
• sloped plywood cover, and
• no methanol
Plot 10 with
• Inoculum at 5 % weight of soil,
• no passive aeration through
corrugated perforated piping,
• no shelter or cover, and
• methanol

Statistic
Average
90% Confidence Interval
Median
Average
90% Confidence Interval
Median
Average
90% Confidence Interval
Median
Average
90% Confidence Interval
Median
Average

90% Confidence Interval

Median

Day 0
77
72-82
77
72
62-82
73
223
190-255
212
190
154-226
196
68

62-74

65

Day 14
89
74- 104
84
80
74-86
81
236
224 - 248
233
227
184-270
217
69

60-78

71

Day28
81
75-87
78
74
67-81
74
249
207-291
236
237
202 - 272
227
77

73-81

76

Day 42
87
83-91
89
81
77-85
81
237
208 - 266
237
246
227 - 265
257
81

73-89

80

Day 56
77
73-81
78
72
68-76
71
225
211 -239
230
230
209 - 251
223
79

72-86

82

-------
Table 4.2-2    Summary of PCB Congener Data for the Various Treatments and Controls

PCB Congener
ID #
1
3
8
15
18
28
37
44
49
52
66
70
74
77
81
87/115
90/1 01
99
110
119
118
123
105
114
126
151
128/167
138/158
149
153/168
156
157
169
170
177
180
183
184
187
189
201
202
194
195
206
207
209
Total Congener
Concentration


Day 0
14
1
1,600
8,000
30,000
57,000
9,200
170
10,000
19,000
14,000
18,000
9,600
1,200
110
2,300
4,500
2,200
4,100
120
3,000
460
2,400
240
19
170
240
1,200
730
560
130
30
ND
130
20
210
49
ND
100
420
38
8
22
10
17
2
5
218,155


Plot # 1
Day 28
11
1
1,600
8,900
14,000
42,000
6,000
16,000
9,800
15,000
14,000
16,000
7,900
1,300
140
2,600
6,000
2,400
4,600
130
3,400
420
2,000
220
14
130
260
1,100
760
580
110
26
ND
100
15
180
40
ND
78
4
28
7
18
9
16
1
4
177,901

PCB Concentration (ug/Kg)

Day 56
7
1
1,200
7,400
13,000
18,000
4,800
15,000
14,000
17,000
14,000
16,000
7,800
1,300
130
2,980
2,400
1,500
2,200
89
2,900
110
1,900
170
18
130
248
1,130
149
901
120
26
ND
100
20
200
42
ND
81
4
4
8
18
10
18
1
4
147,118



Plot # 2
Day 0
15
1
1,600
6,600
27,000
51,000
7,800
14,000
8,800
18,000
12,000
13,000
6,600
980
95
1,500
3,200
1,700
3,300
110
2,600
370
1,800
200
14
100
210
1,000
690
540
110
26
ND
110
16
180
43
ND
92
4
3
7
18
8
13
1
4
185,461

Day 28
12
4
450
3,800
13,000
34,000
6,900
12,000
8,500
13,000
8,500
17,000
15,000
1,500
130
3,100
6,900
2,700
6,600
130
3,100
380
2,300
200
11
86
230
1,100
580
570
110
24
ND
110
15
170
38
ND
70
4
31
6
17
7
17
2
4
162,408

Day 56
13
1
1,400
7,100
1 1 ,000
20,000
5,700
15,000
14,000
17,000
13,000
17,000
7,700
1,400
110
2,990
3,000
1,600
2,300
69
2,500
39
1,500
150
12
120
211
1,110
660
881
98
22
ND
86
16
160
34
ND
63
3
34
7
15
8
16
1
4
148,133

                                        28

-------
Table 4.2-2   Summary of PCB Congener
             (continued...)
Data for the Various Treatments and Controls

PCB Congener
ID #
1
3
8
15
18
28
37
44
49
52
66
70
74
77
81
87/115
90/1 01
99
110
119
118
123
105
114
126
151
128/167
138/158
149
153/168
156
157
169
170
177
180
183
184
187
189
201
202
194
195
206
207
209
Total Congener
Concentration


Day 0
13
1
1,200
5,100
25,000
25,000
6,400
12,000
7,600
14,000
1 1 ,000
110,000
6,700
860
84
1,300
2,600
1,400
2,600
95
2,200
400
1,600
190
13
87
190
750
480
370
97
23
ND
99
15
160
37
ND
74
3
29
7
16
8
12
1
4

141,000

Plot # 3
Day 28
5
1
1,300
8,600
19,000
47,000
74,000
21,000
15,000
23,000
13,000
24,000
21,000
2,000
180
3,800
7,000
3,500
7,500
160
4,800
560
3,300
310
18
140
310
1,500
880
820
170
35
ND
160
23
280
58
ND
110
6
41
10
27
12
28
2
6

238,000
PCB Concentration (ug/Kg)

Day 56 D
9
1
110
6,300
12,000
16,000
6,000
14,000
13,000
16,000
15,000
13,000
7,600
1,300
110
2,860
2,900
1,600
2,200
79
2,400
80
1,700
150
15
120
213
1,070
620
831
98
23
ND
89
16
160
34
ND
64
3
34
7
15
8
15
1
4

138,000


Plot#4D
Day 0
9
1
1,300
5,100
30,000
50,000
7,800
13,000
8,800
16,000
12,000
14,000
7,600
980
37
1,300
2,500
1,400
2,500
77
2,300
490
1,700
180
14
71
190
720
440
330
100
24
ND
96
14
160
38
ND
72
4
31
7
16
8
13
1
4

181,000
Day 28
30
2
4,500
25,000
19,000
50,000
7,700
15,000
1 1 ,000
17,000
9,000
17,000
15,000
1,300
130
3,000
5,400
2,700
5,800
120
3,300
400
2,200
200
14
100
200
1,100
640
570
100
24
ND
110
14
170
38
ND
72
4
31
7
17
8
16
1
4

218,000
Day 56
5
1
1,200
7,100
1 1 ,000
20,000
5,200
14,000
12,000
15,000
15,000
18,000
6,600
1,300
110
3,000
2,800
1,700
2,200
80
3,100
68
1,700
150
15
120
191
1,100
710
871
94
23
ND
79
16
170
36
ND
71
3
29
8
15
9
15
1
4

145,000
                                         29

-------
Table 4.2-2   Summary of PCB Congener
             (continued...)
Data for the Various Treatments and Controls

PCB Congener
ID #
1
3
8
15
18
28
37
44
49
52
66
70
74
77
81
87/115
90/1 01
99
110
119
118
123
105
114
126
151
128/167
138/158
149
153/168
156
157
169
170
177
180
183
184
187
189
201
202
194
195
206
207
209
Total Congener
Concentration


Day 0
9
1
1,400
6,200
36,000
60,000
9,100
17,000
10,000
19,000
14,000
17,000
9,200
1,100
43
1,300
2,800
1,400
2,600
120
2,600
450
1,900
200
16
86
200
860
540
400
110
26
ND
110
17
180
40
ND
81
4
30
7
17
8
15
1
4

216,000

Plot # 5
Day 28
16
1
2,700
17,000
22,000
66,000
12,000
26,000
16,000
2,600
14,000
28,000
24,000
2,200
190
4,000
9,900
3,800
82,000
170
5,100
600
3,100
310
19
140
310
1,500
920
820
160
33
ND
140
22
240
58
ND
100
5
38
10
24
11
28
2
6

272,000
PCB Concentration (ug/Kg)

Day 56 D
6
1
1,500
8,900
13,000
21,000
5,300
12,000
14,000
18,000
16,000
17,000
8,300
1,500
110
3,640
4,300
2,000
2,700
94
3,000
70
2,000
160
17
140
236
1,220
690
1,001
110
26
ND
95
19
180
41
ND
79
3
34
8
18
10
20
1
4

159,000


Plot#6D
Day 0
1
ND
74
1,850
4,600
16,000
2,750
5,550
3,800
3,905
6,000
6,250
3,400
450
28
555
1,250
590
1,200
26
1,200
110
795
58
5
68
105
560
315
255
57
14
ND
59
8
91
21
ND
41
2
10
2
8
3
6
0
2

62,000
Day 28
1
ND
160
7,450
2,700
15,000
2,850
6,350
4,800
7,350
4,150
8,000
7,000
630
57
1,300
3,250
1,200
2,800
61
1,700
180
1,020
92
7
61
155
730
430
395
74
16
ND
71
9
105
23
ND
39
3
16
3
9
4
8
1
2

78,000
Day 56
0
ND
55
2,000
2,100
7,450
2,750
8,250
6,550
8,200
7,600
8,500
4,350
705
59
1,480
1,600
770
1,305
313
1,750
44
1,100
78
9
87
173
920
475
691
84
19
ND
71
13
130
29
ND
23
3
21
5
10
6
9
1
2

68,000
                                         30

-------
Table 4.2-2   Summary of PCB Congener
             (continued...)
Data for the Various Treatments and Controls

PCB Congener
ID #
1
3
8
15
18
28
37
44
49
52
66
70
74
77
81
87/115
90/1 01
99
110
119
118
123
105
114
126
151
128/167
138/158
149
153/168
156
157
169
170
177
180
183
184
187
189
201
202
194
195
206
207
209
Total Congener
Concentration


Day 0
ND
ND
22
1,300
3,100
13,000
2,400
3,900
2,700
4,800
5,200
3,700
2,200
390
28
370
900
470
930
22
850
90
660
48
4
53
80
460
270
220
45
10
ND
46
6
70
16
ND
31
2
7
2
6
3
4
ND
1

48,000

Plot # 7
Day 28
ND
ND
45
2,900
1,500
1 1 ,000
2,500
5,200
4,000
6,200
3,500
6,600
6,000
520
54
1,200
2,800
1,000
2,200
55
1,500
140
860
75
6
44
120
610
380
320
58
13
ND
57
7
82
16
ND
32
2
16
3
7
3
8
1
2

62,000
PCB Concentration (ug/Kg)

Day 56 D
ND
ND
32
1,900
1,200
3,600
2,000
4,600
4,500
4,200
5,200
4,500
1,900
510
34
1,050
1,000
670
800
29
1,100
29
680
51
6
63
115
710
410
541
53
12
ND
47
8
85
18
ND
1
2
12
3
6
4
6
ND
1

42,000


Plot#8D
Day 0
13
1
1,600
7,800
26,000
54,000
7,400
13,000
8,000
15,000
1 1 ,000
14,000
7,400
1,000
120
1,000
2,200
1,000
2,100
60
2,500
210
2,000
180
9
120
160
940
570
440
90
20
ND
97
15
160
37
ND
76
3
23
5
16
7
14
1
4

180,000
Day 28
4
1
1,400
8,000
20,000
44,000
7,300
18,000
13,000
20,000
9,800
19,000
17,000
1,500
120
2,800
7,000
2,600
5,700
120
3,400
400
2,300
200
13
100
210
1,100
670
600
110
23
ND
110
16
170
39
ND
81
4
28
7
18
8
16
1
4

207,000
Day 56
2
ND
780
4,000
22,000
19,000
6,300
18,000
22,000
15,000
22,000
16,000
1 1 ,000
1,500
96
2,240
2,000
1,400
1,700
77
2,700
57
1,900
210
12
120
201
1,088
630
881
94
23
ND
81
17
160
37
ND
70
3
28
7
15
9
16
1
3

173,000
                                         31

-------

PCB Congener
ID #
1
3
8
15
18
28
37
44
49
52
66
70
74
77
81
87/115
90/1 01
99
110
119
118
123
105
114
126
151
128/167
138/158
149
153/168
156
157
169
170
177
180
183
184
187
189
201
202
194
195
206
207
209
Total Congener
Concentration


Day 0
14
1
1,200
5,800
27,000
56,000
7,900
14,000
8,100
16,000
12,000
15,000
8,300
1,000
40
1,200
2,500
1,300
2,300
86
2,900
400
2,100
180
13
160
210
920
600
460
100
25
ND
100
16
180
40
ND
78
4
26
6
18
7
15
1
4

188,000

Plot # 9
Day 28
4
1
1,700
9,800
21,000
48,000
7,100
18,000
13,000
20,000
9,900
20,000
16,000
1,500
140
3,200
7,600
2,900
6,100
140
3,600
440
2,200
210
12
98
220
1,200
740
640
110
24
ND
110
17













216,000
PCB Concentration (ug/Kg)

Day 56
2
ND
710
4,000
22,000
15,000
6,500
30,000
24,000
22,000
23,000
18,000
13,000
1,600
99
2,000
2,100
1,300
1,600
86
2,800
58
1,800
150
14
130
202
1,090
620
891
98
24
ND
82
18
170
39
ND
75
3
28
7
16
9
16
1
4

195,000


Plot* 10
Day 0
ND
ND
44
2,600
4,700
21,000
3,800
6,400
4,000
7,300
7,000
6,900
4,000
540
45
530
1,200
600
1,200
46
160
20
1,100
99
7
100
150
720
410
300
83
19
ND
75
11
120
26
ND
50
3
16
4
10
5
8
1
2

75,000
Day 28
ND
ND
46
2,600
2,000
1 1 ,000
2,200
5,500
3,900
6,100
3,200
6,300
5,500
520
52
1,000
2,500
900
2,000
50
1,300
150
800
72
6
45
120
570
340
310
61
13
ND
56
7
81
18
ND
33
2
12
3
7
3
8
1
2

59,000
Day 56
ND
ND
31
1,400
2,800
6,400
2,000
6,900
5,000
4,600
5,900
4,600
3,400
550
36
840
880
670
720
29
1,100
23
710
56
7
64
117
590
310
471
54
12
ND
45
8
82
18
ND
31
2
12
3
6
4
6
0
1

50,000
32

-------
Table 4.2-3   Summary of PCB Homologue Data for the Various Treatments and Controls
                                                       PCB Concentration (ug/Kg)
PCB
Homologue
Total monoCB
Total diCB
Total triCB
Total tetraCB
Total pentaCB
Total hexaCBI
Total heptaCB
Total octaCB
Total nanoCBI
Total Homologues
Plot # 1
Day 0 Day 28 Day 56
16 12 8
17,000 18,000 22,000
190,000 120,000 90,000
150,000 130,000 140,000
29,000 31,000 21,000
4,000 3,700 3,900
720 580 670
120 96 110
24 22 25
391,000 303,000 278,000
Plot # 2
Day 0 Day 28 Day 56
17 17 15
16,000 14,000 15,000
170,000 110,000 97,000
130,000 120,000 140,000
24,000 35,000 21,000
3,500 3,400 3,600
610 560 540
99 98 99
19 24 22
344,000 283,000 277,000
Plot # 3
Day 0 Day 28 Day 56
14 5 11
13,000 17,000 13,000
150,000 140,000 97,000
100,000 150,000 140,000
21,000 54,000 20,000
2,700 4,900 3,500
540 900 560
94 1 40 99
17 38 20
287,000 358,000 274,000
Plot # 4
Day 0 Day 28 Day 56
10 35 5
12,000 48,000 12,000
170,000 140,000 95,000
120,000 130,000 130,000
20,000 32,000 22,000
2,500 3,400 3,700
540 550 570
98 100 94
18 22 20
325,000 354,000 263,000
Plot # 5
Day 0 Day 28 Day 56
11 18 7
14,000 31,000 15,000
180,000 180,000 110,000
150,000 220,000 140,000
23,000 50,000 25,000
3,000 4,800 4,000
600 820 640
96 130 110
20 37 260
371,000 487,000 295,000
PCB
Homologue
Total monoCB
Total diCB
Total triCB
Total tetraCB
Total pentaCB
Total hexaCBI
Total heptaCB
Total octaCB
Total nanoCBI
Total Homologues
Plot # 6
Day 0 Day 28 Day 56
1 1 0
3,900 6,250 2,450
38,000 37,000 27,500
45,000 57,000 70,500
8,000 16,500 11,350
1,800 2,350 2,850
310 345 455
30 49 64
8 11 12
97,000 120,000 115,000
Plot # 7
Day 0 Day 28 Day 56
ND ND ND
1,800 3,600 2,200
29,000 28,000 20,000
36,000 48,000 39,000
6,300 14,000 7,400
1,500 1,900 2,200
240 280 290
25 45 38
6118
75,000 96,000 71,000
Plot # 8
Day 0 Day 28 Day 56
15 4 2
17,000 17,000 9,200
160,000 120,000 130,000
110,000 280,000 150,000
17,000 33,000 19,000
3,000 3,500 3,600
540 600 560
78 93 87
19 23 21
308,000 454,000 312,000
Plot # 9
Day 0 Day 28
15
15,000
1 70,000
130,000
20,000
3,300
580
88
21
339,000
Day 56
2
9,100
130,000
210,000
19,000
3,600
590
90
22
372,000
Plot* 10
Day 0 Day 28 Day 56
ND ND ND
3,500 3,300 1,800
54,000 28,000 26,000
56,000 47,000 45,000
11,000 13,000 6,600
2,400 1,800 1,900
390 270 280
53 39 36
10 10 8
127,000 93,000 82,000

-------
                                    Section 5.0
                             QUALITY ASSURANCE
5.1      QA Summary
Quality Assurance (QA) may be defined as a system of activities the purpose of which is to provide
assurance that defined standards of quality are met with a stated  level of confidence.  A QA
program is a means of integrating the quality planning, quality assessment, quality control (QC),
and quality improvement efforts to  meet user requirements.  Included are all actions taken by
project personnel, and the documentation of laboratory and field performance. Typically, project-
specific QA/QC requirements are specified in a Quality Assurance  Project Plan (QAPP). The
objective of the quality assurance program is to reduce measurement errors to agreed upon limits
and to produce results of acceptable and known quality.  The QAPP specifies the necessary
guidelines to ensure that the measurement systems remain  in control and provides detailed
information on  the analytical approach to ensure that data of  acceptable quality is obtained to
achieve project objectives.

For the preliminary evaluation of the GML PCB Bioremediation Process, a Treatability Test Plan
(TP), instead of a QAPP, was developed, approved and implemented.  The following  sections
provide information on the use of data quality indicators, limitations on data use and a summary of
the QC analyses associated with  project measurements.
5.2     Data Quality Indicators

To assess the quality of the data generated for this field test, two important data quality indicators
are of primary concern: precision and accuracy. Precision can be defined as the degree of mutual
agreement of independent measurements generated through repeated application of the process
under specified conditions. Accuracy is the degree of agreement of a measured value with the true
or expected value.

Precision is generally measured by laboratory/matrix spiked sample duplicates and field sample
duplicates. In the case of duplicates, precision is evaluated by expressing, as a percentage, the
difference between results of the sample and sample  duplicate results.   The relative percent
difference (RPD) is calculated as:

             RPD = (Maximum Value-Minimum Value) x 100n
                    (Maximum Value+Minimum Value)/20

For three or more measurements, precision is evaluated by the standard deviation of the multiple
measurements relative to the mean, i.e. the  relative standard deviation (RSD), according to the
following equation:

             RSD= (SD/Xavg) x100

Where SD is the standard deviation and Xavg  is the average of the multiple concentrations.
                                         34

-------
To determine and evaluate accuracy, known quantities of select target analytes were spiked into
selected field samples.  Equipment used to provide data for this project was tested for accuracy
through the analysis of calibration check standards and laboratory control samples. To determine
matrix spike recovery, the following equation was applied:

             % Recovery = Css^s x 1 00
where   Css   =      Analyte concentration in spiked sample
        Cus   =      Analyte concentration in unspiked sample
        Csa   =      Analyte concentration added to sample
To determine the % recovery of LCS analyses or spiked blanks, the equation below was used:

             % Recovery = Measured Concentration    x 100D
                          Theoretical Concentration n

Another important aspect of assessing data quality is completeness. Completeness is a measure
of the amount of valid data produced from the total effort compared to the total amount of data
determined to be necessary to meet project objectives.

To determine if a measurement is valid, it must be  reproducible and comparable. Comparability
expresses the extent to which one data set can be compared to another. To generate comparable
results, standard methods that are widely accepted along with strict analytical and field protocols
were used. These methods were clearly specified in the TP and reviewed before samples or data
were collected.

While several precautions were taken to generate data of known quality through the control of the
measurement system, the data must also be representative of true conditions. Representativeness
refers to the degree with which analytical  results accurately and precisely reflect actual conditions
present at the locations chosen for sample collection.
5.3     Conclusions and Data Quality Limitations

A review of the critical sample  data and associated QC analyses was performed to determine
whether the data collected were of adequate quality to provide proper evaluation of the project's
technical objectives. The only critical measurement forthis technology demonstration was the PCB
Aroclor analysis of the soil grab samples from the various treatment and control experiments.
These samples were  collected on Days 0,  14, 28 42 and 56 of the field treatability testing. The
results of the measurements designed to assess the data quality objectives for these analyses are
summarized and discussed below.

Accuracy for the analysis of PCB Aroclor in the soil  grab samples collected from  the various
treatment and  control plots was assessed  by the analysis of Matrix Spike Duplicate Samples
(MS/MSD).  As it turned out, the predominant PCB constituent in the target soil  matrix was Aroclor
1248. Therefore, the MS/MSD samples were spiked with Aroclor 1248. For the most part (i.e., for
22 out of the 24 pairs of MS/MSDs) the critical compounds met the data quality objectives with
average % recovery values ranging between 71 -123%, which was within the established control
limits of 70 - 130 % for the Aroclor analysis.

-------
Precision was evaluated through the Aroclor 1248 analysis of field duplicate samples as well as
laboratory paired matrix spiked duplicates (MS/MSDs).  For the most part (i.e., for 22 out of the 25
pairs of field duplicate samples) the % RPD values were within specified control limit of 30%, with
an average % RPD of 16.3.  For the most part (i.e.,  for 22 out of the 24 pairs of laboratory
MS/MSDs samples) the % RPD values were within specified control limit of 30%, with an average
% RPD of 19.7.

Detection limits as reported met objectives as stated in  the TP.

Comparability was achieved through the use of EPA approved analytical methods and protocols
and verified by the validation of analytical data, which indicated that most TP and method-specified
criteria were met.

Completeness objectives were met for the treatability study phase sampling and analytical program.
                                          36

-------
                                  Section 6.0

                                 REFERENCES
SITE Program - Test Plan (July 1998) for Treatability Study of Green Mountain Laboratories, Inc.'s
       Bioremediation Process for Treatment of Soils Contaminated with PCBs at the Beede
       Waste Oil/Cash Energy Superfund Site, Plaistow, New Hampshire.

Rutkowski, Anthony A., U.S. Patent - 6,096,531, August 1, 2000, Methods and composition for
       bioremediation.
                                       37

-------